Advances in Animal Science Volume II

Advances in Animal Science
Volume II
(ISBN: 978-93-91768-45-4)
Editors
Dr. Gyanendra Kumar Dr. Anil Madhukarrao Khole

Department of Zoology, Department of Zoology,
National P. G. College, B. Raghunath Arts, Commerce
Lucknow (U.P.) and Science College, Parbhani (M.S.)
Mr. Debajyoti Pradhan Dr. Naveen Kumar Chourasia

Department of Zoology, Department of Zoology,
Egra Sarada Shashi Bhusan College Govt. College Bichhua,
(under Vidyasagar University), Chhindwara
Egra (W. B.)

ISBN: 978-93-91768-45-4
 Copyright reserved by the publishers
Publication, Distribution and Promotion Rights reserved by Bhumi Publishing, Nigave Khalasa, Kolhapur
Despite every effort, there may still be chances for some errors and omissions to have crept in
inadvertently.
No part of this publication may be reproduced in any form or by any means, electronically, mechanically,
by photocopying, recording or otherwise, without the prior permission of the publishers.
The views and results expressed in various articles are those of the authors and not of editors or
publisher of the book.
Published by:
Bhumi Publishing,
Nigave Khalasa, Kolhapur 416207, Maharashtra, India
Website: www.bhumipublishing.com
E-mail: [email protected]
Book Available online at:
https://www.bhumipublishing.com/books/
PREFACE
We are delighted to publish our book entitled "Advances in Animal Science Volume
II". This book is the compilation of esteemed articles of acknowledged experts in the fields
of basic and applied animal science.
This book is published in the hopes of sharing the excitement found in the study of
animal science. Animal science can help us unlock the mysteries of our universe, but
beyond that, conquering it can be personally satisfying. We developed this digital book
with the goal of helping people achieve that feeling of accomplishment.
The articles in the book have been contributed by eminent scientists, academicians.
Our special thanks and appreciation goes to experts and research workers whose
contributions have enriched this book. We thank our publisher Bhumi Publishing, India for
taking pains in bringing out the book.
Finally, we will always remain a debtor to all our well-wishers for their blessings,
without which this book would not have come into existence.
- Editors
Advances in Animal Science Volume II
CONTENTS
Sr. No. Book Chapter and Author(s) Page no.
1. FORMULATION OF FISH FEED USING TRADITIONAL MEDICINAL 1 – 12

PLANT VETIVERIA ZIZANIOIDES FOR TREAT EPIZOOTIC
ULCERATIVE SYNDROME INFECTED ORNAMENTAL FISH KOI
CARP - CYPRINUS CARPIO
R. Bhuvaneswari and N. Manickam
2. POTENTIAL OF WHEATGRASS BASED FORMULATED FEED IN 13 – 29

FRESH WATER FISH CULTURE
S. S. Butle, R. A. Gulhane and P. S. Joshi
3. PRODUCTION OF VACCINE AGAINST GASTROINTESTINAL 30 – 36

NEMATODE HELIGMOSOMOIDES POLYGYRUS
Dipti Bhimrao Kadu
4. TECHNOLOGICAL ADVANCES AND ANIMAL HEALTH 37 – 42

Mohsina Mushtaq
5. CLONING AND GENETIC ENGINEERING 43 – 49

Khan Rumana Amanullah
6. EMERGING TOOLS FOR CONSERVATION AND 50 – 54

MONITORING OF BIODIVERSITY
Sachin K. Shelake and Vishwas Y. Deshpande
7. DIMETHOATE: KILLER PESTICIDE 55 – 56

Chandani B. Jamdade
8. DIVERSITY OF MOTHS (LEPIDOPTERA: HETEROCERA) AND 57 – 62

THEIR POTENTIAL ROLE AS A CONSERVATION TOOL IN
PROTECTED AREAS OF KATEPURNA WILDLIFE SANCTUARY
OF AKOLA (M.S.)
Priyanka M. Ramteke
9. CLIMATE CHANGE AND ITS INTERACTIONS WITH PEOPLE 63 – 68
AND ECOSYSTEMS
Ansari Abdullah Daniyal
10. SERICULTURE BASED INTEGRATED FARMING SYSTEM FOR 69 – 76

SUSTAINABLE AGRICULTURAL DEVELOPMENT
Suraksha Chanotra and Muzafar Ahmad Bhat
11. AN OVERVIEW OF DIFFERENT TYPES OF TOXIC 77 – 89

ENVIRONMENTAL CHEMICALS AND THEIR
CELLULAR RECEPTORS
Deeksha Sharma
12. HEAT STRESS AND ITS IMPACT ON HORMONAL PHYSIOLOGY 90 – 99

OF HOMEOTHERMIC ANIMALS
Manomita Maity and Debajyoti Pradhan
13. DETERMINATION OF MICROBIAL GUT DENSITY IN 100 – 108

FRESH WATER FISHES
Annapurneshwari H
14. STUDIES ON BREEDING CHARACTERISTICS OF NATURALLY AND 109 – 121

INDUCED BRED DOTTYBACK FISH,
PSEUDOCHROMIS DILECTUS (LUBBOCK, 1976)
Nanthini devi Karuppaiya, Santhanam Perumal and
Perumal Pachiappan
15. UNDERSTANDING GENETIC 122 – 123

Vinda Manjramkar
16. VERMICOMPOSTING OF RICE STRAW USING EARTHWORM 124 – 129

EUDRILUS EUGENIAE
Sivasankari. B, Priyadharshini. R and Ganesh. S
17. DICHLORVOS MEDIATED BEHAVIOURAL RESPONSESIN FRESH 130 – 134
WATER FISH, CHANNA GACHUA (F. HAMILTON)
P. S. Pawar and R. N. Patil
18. VERMICOMPOSTING OF SUGAR CANE BAGASSE USING 135 – 139

EISENIA FETIDA
Sivasankari B, Nathiya G. and Ganesh S
19. BLACK SOLDIER FLY, HERMETIA ILLUCENS L.: A NUTRITIVE 140 – 146
INSECT AND A SOLUTION TO LIVESTOCK FEED
Karuna P. Ganvir
20. AN ASSESSMENT OF INVERTEBRATE SPECIES IN 147 – 153

WETLAND ECOSYSTEMS
Wasudha J. Meshram
(ISBN: 978-93-91768-45-4)
FORMULATION OF FISH FEED USING TRADITIONAL MEDICINAL PLANT

VETIVERIA ZIZANIOIDES FOR TREAT EPIZOOTIC ULCERATIVE SYNDROME
INFECTED ORNAMENTAL FISH KOI CARP - CYPRINUS CARPIO
R. Bhuvaneswari*1 and N. Manickam2
1Fish Disease Diagnostic Laboratory, Department of Zoology,
AyyaNadar Janaki Ammal College, Sivakasi-626 124, Tamil Nadu, India
2Marine Planktonology and Aquaculture Laboratory, Department of Marine Science,
Bharathidasan University, Tiruchirappalli-620 024, Tamil Nadu, India
*Corresponding author E-mail: [email protected]; [email protected]
Abstract:
Nowadays, our focus completely towards herbs and their medicinal usages, reasons are
low cast easily available low side effects and environmentally friendly he world population,
especially in developing countries depends on the traditional system of medicine for a variety of
diseases. The medicinal plants can be used not only against diseases but even more so, as growth
promoters, stress resistance boosters and preventatives of infections. Additionally,
phytomedicines provide a cheaper source for treatment and greater accuracy than
chemotherapeutic agents without causing toxicity. The herbs can also act as immunostimulants,
conferring the non-specific defense mechanisms of fish and elevating the specific immune
response. In the present article, traditional medicinal plant Vetiveria zizanioides added as a major
ingredient in fish feed formulation whichacting against EUS fish diseases, the root powder of V.
zizanioides containing feedwas fed to experimentally infected EUS (2.7 × 104CFU) Ornamental
Koi fish Cyprinus carpio maintained in laboratory cement tanks where, 98±2.35% were found to
be recovered at 20th day. Tetracycline powder added feed used as a control in this study showed
medium to weak effect recovering the EUS infected fish.
Key words: Vetiveria zizanioides, Epizootic ulcerative syndrome, Ornamental Koi, Cyprinus
carpio, Herbal feed, Tetracycline feed, Treatment
Introduction:
Epizootic Ulcerative Syndrome (EUS) is one of the most destructive diseases among
fresh and brackish water fish in the Asia-Pacific region and it is very common in northern and
southern India, causing considerable loss to fish farmers (Chinabut et al., 1995). The emergence
1
Bhumi Publishing, India
and spread of aquatic freshwater diseases are a major conservation concern. EUS is caused by
the fungus-like oomycete Aphanomyces invadans and Aeromonas hydrophila can cause
significant ulceration of the skin, necrosis of muscle with extension to subjacent structures
including abdominal cavity and cranium, and leading to mortality in many cases.India witnessed
the first major outbreak of EUS in 1988 in the states of Tripura, Assam, Meghalaya and West
Bengal. It gradually spread until 1992 in the states of Orissa, Bihar, Uttar Pradesh, Madhya
Pradesh, Maharashtra, Andhra Pradesh, Tamil Nadu, Kerala, Karnataka and Haryana (Das,
1997). In India, Central Inland Fisheries Research Institute (CIFRI) has been monitoring the
disease since early 1988. Over the past two decades, EUS has had a serious impact on tropical
fisheries, resulting in heavy economic loss. It is one of the most destructive diseases in fresh and
brackish water fish in the Asian Pacific region. It is very common in India and has spread
through rivers, reservoirs and paddy fields to neighboring states, causing considerable loss to fish
farmers (Muthukrishnan et al., 2008). Vetiver known as Khus grass is a perennial grass of Indian
origin. In India it is mainly used in ayurvedic medicine. The plant has This property of Vetiver
Essential Oil helps heal wounds by promoting growth of newtissues at the wounded place and
also by keeping it safe from infections by inhibiting growth of microbes and promoting crowding
of leucocytes and platelets at the place.Vetiver helps in the formation of new tissue is used so as
to accelerate the healing and recovery of skin wounds as well to remove stains, marks on the skin
and the scars themselves. Also, we used to repair the cracks and grooves in the skin caused by
different circumstances such as pregnancy, diets, allergies, burns.
Many fish farmers and ornamental fish hobbyists buy the bulk of their feed from
commercial manufacturers. Small ornamental fish farms with an assortment of fish require small
amounts of various diets with particular ingredients. However, the small quantities of fishfeeds
can be made quite easily in the laboratory, or at home, with common herbal ingredients and with
simple kitchen or laboratory equipment. Henceforth, this chapter will provide the knowledge of
herbal feed formulation, which are beneficial for fish health and may act against EUS disease
causing pathogens A. invadans and A. hydrophila. From our earlier study it was found that EUS
causing microbesA. invadans and A. hydrophila 3.7 × 104 CFU completely ward off by the
herbal extract of V. zizanioides in vitro study Based on the findings, the present study was
designed to evaluate the efficacy of V. zizanioides in healing the lesion caused by EUS disease
causing pathogens A. invadans and A. hydrophila.
Materials and method:

Plant material:
The medicinal plant was selected for the study from the local area based on their basic
2
(ISBN: 978-93-91768-45-4)
information in the available. The medicinal plant Vetiveria zizanioides were collected from
follow land in and around Orathanadu, Thanjavur brought into the laboratory for further
processes. The collected samples were carefully stored in sterile polythene bags and used for the
further study. V. zizanioides are widely distributed in tropical and warm temperate regions. These
plants are commonly used in folk medicine to treat dermatitis, gastric ulcer, abrasion, lesions and
inflammation (Vaidyarathnam, 1995).
Preparation of plant powder:
The fresh rhizome of V. zizanioides was collected and washed under running tap water
for 5 minutes; the small hairs of the rhizome were removed and the parts of the plants were
chopped and shade dried for a week to achieve weight constancy. The dried parts were finely
powdered in an electric blender and stored in airtight bottles
Experimental animal:
The Koi carp, Cyprinus carpio (27.6±4.2 g) obtained from local ornamental fish farm in
Kadachanendhal, Madurai, Tamil Nadu, were used in the experiments. The fish were transported
to the laboratory in plastic bags (5L) filled with oxygenated water. They were acclimatized in
cement aquaria (capacity 2000L; 6×4×4 m) for 3 weeks to laboratory conditions (10 hrs. dark: 14
hrs. light). The observed water quality parameters are: dissolved oxygen.
Pathogen: Aeromonas hydrophila:
The reference strain A. hydrophila (MTCC code no-646) was purchased from the
Institute of Microbial Technology (Government of India), Chandigarh and maintained in the
laboratory under standard conditions. Subcultures were maintained on Tryptone soya agar slopes
at 30˚ C (Hi-Media, Mumbai) (Joseph and Carnahan, 1994).
Fungi isolation and identification:
Fungi species were isolated from the water samples by baiting method and using
sterilized maize, and hemp seed as described by Stevens (1974). To induce sporulation, place an
agar plug (3-4 mm in diameter) of actively growing mycelium in a Petri dish containing GPY
broth and incubate for 4 days at approximately 20°C, after about 12 hours, the formation of
achlyoid clusters of primary cysts and the release of motile secondary zoospores should be
apparent under the microscope.
Infective experiment:
Fish can be experimentally infected by intramuscularly injecting a 0.1 ml suspension of
100+ motile zoospores into fish susceptible to infection with A. invadans and with A. hydrophila
at the concentrations of 104CFU/ml at 28°C, and demonstrating histological growth of aseptate
hyphae, with red spot at 14mm in diameter, in the muscle of fish sampled after 3rd days. Each
3
experiment was carried out in triplicate with 10 fishes in each to quantify survival rate and ID 50
values. The experiment lasted for 45 days. Every day the size of the induced lesion was
measured with vernier caliper.
Intramuscularly injective experimental design:
Appropriate volume (0.025ml/100g body weight) of the selected serial dilution
(101,102,103,104,105or 106CFU/fish) was injected intramuscularly beneath the dorsal fin of the
fish using a tuberculin syringe using 20-gauge needle size (Scharperculeus, 1991). For each
dilution a triplicate with ten fishes were kept and the fishes (n=10) were kept in plastic aquaria
(capacity 10L). Mortality was monitored continuously on each days and dead fish, if any,
removed with a forceps. The fish was considered dead when gill movement ceased and there was
no response to gentle prodding (Sprague, 1973). The medium was not changed or aerated during
this period (Peer Mohamed, 2000).
Confirmation of pathogenesis:
The dose which afforded at least 20% survival of the infected fish beyond 12 days after
infection selected to observe to pathological signs. The following clinical signs were used to
confirm the induction of pathogenesis: a) discoloration of the body and the gills, b) abnormal
swimming-rushing around, whirling, reeling, rotating or swimming upside down, c) refusal of
food, d) retarded movement to the water surface or bottom, e) loss of equilibrium, f) closed fin-
dorsal fin, g) swelling of abdomen, h) erection of scales, and i) appearance of external bumps,
sores or lesions (Balasubramanian, 2005). The cause of the disease in the fishes, which survived
the inoculation, was confirmed by isolation of bacteria from the outer edge of the skin lesions.
Sterile nutrient broth was inoculated with the swabs from the skin lesions. The bacterial cells
were harvested and their characteristics compared for their identity to A. hydrophila (Selvaraj et
al., 2004).
Feed preparation:
The fish feed was prepared by mixing the selected feed ingredients. The mixture of the
feed ingredient was moistened with water and steam cooked in batches and cooled. To this feed,
vitamin mix and cod liver oil capsule were also added, and then mixed thoroughly to make
Dough. This feed mixture was pelletized through hand pelletized until a size of 1.0 to 4mm and
dried in shade to reduce the moisture content of the feed below 10% (Vijayakumari and
Sukumaran, 2014).
Experimental feed preparation:
The medicated feed was prepared by using the V. zizanioides40gm herbal plant powder,
15gm groundnut oil cake, 15gm fishmeal, vitamin c capsule 2mg, rice bran 5gm, tapioca powder
5 gm, cod liver oil capsule 2mg. The control feed was prepared by using the tetracycline40gm
4
(ISBN: 978-93-91768-45-4)
herbal plant powder, 15gm groundnut oil cake, 15gm fishmeal, vitamin c capsule 2mg, rice bran
5gm, tapioca powder 5 gm, cod liver oil capsule 2mg. The normal diet was prepared by using
without plant powder, 15gm groundnut oil cake, 15gm fishmeal, vitamin c capsule 2mg, rice
bran 5gm, tapioca powder 5 gm, cod liver oil capsule 2mg.
Results:
Infective dosage:
In C. carpio, A. hydrophila with A. invadans infection produced mean percentage
mortality 87, 73, 50, 44, 40 and 20 at (Group I) 3.9×101, (Group II) 3.5×102, (Group III)3.1×103,
(Group IV) 2.7×104, (Group V) 2.3×105, and (Group VI) 1.9×106 CFU/ml concentrations
respectively. After 15 days infection produced 50% mortality with 2.7×104CFU/ml (Table 1).
Since 50% mortality was observed at 2.7×104CFU /ml concentration. This dose was chosen for
all the experiments.
Plate I: Pathogens culture

Table 1: Decrease in the lesion size (14mm) of infected C. carpio treated withV. zizanioides
powder added /not added during 15 days
Group 3rd day 5th day 10th day 15th day 20th day
1.42±0.59 1.72±0.2 2.4±0.07 3.9±0.03 4.6±0.05
Infected untreated
(14mm) (17mm) (24mm) (39mm)
Infected fed with V. zizanioides 1.42±0.59 0.53±0.33 0.39±0.31 0.22±0.00 0.00±0.00
powder added feed (14mm) (05mm) (03.1mm) (02.2mm)
1.42±0.59 1.73±0.3 2.38±0.17 3.92±0.06 4.1±0.03
Infected fed with normal feed
(14mm) (17mm) (23mm) (039mm) (41mm)
Infected treated with 1.42±0.59 1.62±3.3 1.53±0.29 1.51±0.07 1.31±0.07
Terramycin (14mm) (16.2mm) (15.3mm) (15.1mm) (13.1mm)
5
Clinical Sign and Gross Pathology:

In general, lesions on EUS-affected fish can be separated into three groups on the basis of
gross appearance. Clinical signs in the early stage of the disease are similar. Appetite is reduced
or absent and fish become lethargic, either floating just beneath the surface or swimming with
the head out of the water Pinhead-sized, red spots develop on the body surface, head and fins,
caudal peduncle, dorsum or operculum with no noticeable hemorrhages or ulcers. The
intermediate stage lesions are represented by small (1.42 cm) dermal ulcers, with associated loss
of scales, hemorrhages and oedema.
Table 2: Clinical sign and gross pathological funding in C. carpio (Ornamental Koi carp)
infected with A. invadans and A. hydrophila at the concentrations of 2.7 x 104CFU/ml
Clinical sign and pathological gross
12 hrs. Abnormal swimming
24 hrs. Abnormal swimming-rushing around, whirling, reeling,
rotating or swimming upside down
nd
2 Day Refusal of food, loss of equilibrium
rd
3 Day Pinhead-sized, red spots develop on the body surface
Duration
5th Day Noticeable hemorrhages or ulcers
7th Day Dermal ulcers, with associated loss of scales,
9th Day Closed fin-dorsal fin, resulting eventually in death
11th Day large necrotic open ulcers; resulting eventually in death
13th Day large necrotic open ulcers; resulting eventually in death
The advancedstage lesions appear on other parts of thefish‘s body and expand into large
necrotic open ulcers; resulting eventually in death. Some affected species, such as striped
snakehead, can survive with much more severe, chronic lesions that may have completely
destroyed the caudal peduncle or eroded into the cranium or abdominal cavity, sometimes
exposing the swim bladder (Table 2).
Size of the lesion:
The size (mm) of the lesion increased as a function of infection period (25days). The
mean size of lesion on 3rd day was 1.02±0.01 mm and on the 15th day increased into
9.29±0.10mm on the in the ideal dosage 2.7×104CFU/ml (Table 1). In I group high concentration
of infected with A. invadans and A. hydrophila 3.7×101CFU the mean size of lesion increased
from 3.2±0.02 mm on the 3rd day to 9.88±0.06mm on the 20th day and 90% of mortality occurred
on the 20th day in this concentration. Approximately the pathogens are present in the
6
(ISBN: 978-93-91768-45-4)
concentration was 8.15 million /ml and in the ID50 concentration 6.45 million /ml. Infected fishes
the mean size of lesion at the least concentration of 2.06×106CFU/ml caused from 1.50 ± 0.0 mm
on the 3rd day to 4.62±3.17mm on the 15th day. Infected fed with V. zizanioides powderadded
feed showed complete healing on 20th day (Plate-I)
Plate II: Infected Cyprinus carpio treated with Vetiveria zizanioides powder added feed
Mortality:
On 30th day of infection 90 % of mortality occurred at the high concentration of A.
hydrophila 3.7×104CFU/ml. The lesion 9.29±0.10mm on 15th day in the ideal dosage
2.7×104CFU/ml while on 30th day fish suffered 50 % mortality. In the infected fishes, the mean
mortality value 20 % on 30th day at the least concentration of 2.06×106CFU/ml. But mortality
rate completely nil in infected fish fed and treated with V. zizanioides added feed.
Preparation of plant powder:
The dried V. zizanioides were grinded into powdered form mechanically and sieved with
a hand sieve to obtain 500 g of each of the powders which were then stored in airtight containers
till formulation and preparation of experimental diets. Fresh weight was 500gm, dried weight
was 462.7gm.
7
Plate III: Dried V. zizanioides

Feed formulation:
Experimental diet was formulated using the Pearson‘s square method and distributed into
treatments. Feed ingredients used for the experimental diets includeV. zizanioides 40gm herbal
plant powder,15gm groundnut oil cake, 15gm fishmeal, sodium chloride 1mg,rice bran 5gm,
tapioca powder 5 gm, cod liver oil capsule 2mg.The formulation was based on the percentage
composition of the ingredient (Table 3). To prepare the diets, the dried and grinded ingredients
of each diet were weighed and mixed thoroughly in a bowl, vegetable oil and warm water were
then added slowly along to the mixture and mixed manually for about 30 minutes to achieve a
proper consistency. The resulting mixture was pelletized (2 mm) using a traditional pelletizerand
allowed to dry for 24 hrs, by air circulation before being packed into airtight containers and
stored at room temperature to be crumbled before use. Formulated diet samples (10 g) were
analyzed following the procedures of AOAC. Moisture was analyzed by drying the sample in an
air convection oven at 105oC overnight.
Table 3: Formulation and proximate composition of experimental diets (40g/100 g dry
weight)
Ingredients Experimental diet Normal diet Tetracycline diet
V. zizanioides 40 gm Nil Nil
Fish Meal 15 gm 15 gm 15 gm
Soybean cake 10 gm 10 gm 10 gm
groundnut oil cake 15 gm 15 gm 15 gm
Rice bran 5 gm 5 gm 5 gm
Tapioca flour 5 gm 5 gm 5 gm
Veg.oil 3 ml 3 ml 3 ml
Sodium chloride 1 mg 1 mg 1 mg
Cod liver oil capsule 1 mg 1 mg 1 mg
Tetracycline Nil Nil 40 gm
8
(ISBN: 978-93-91768-45-4)
Plate IV: Herbal feed
Discussion:
The LD50 value of Clarias batracus (ip) injected with A. hydrophila was 3.79 x
1010CFU/ml at 96 hrs (Supap, 1985). Experimentally infected Oreochromis with A. hydrophila
showed LD50 value of 1.5 106 at 96 hrs (Yambot, 1998). The isolates of A. hydrophila and A.
sobria showed LD50 value of 103 and 104 respectively indicating their virulence in native fish
(Shariff and Subasinghe, 1994). In rainbow trout (Salmo gairnern) when 5×106CFU/ml were
injected intramuscularly; the bacterial cells were completely cleared from the site of the injection
and the organs; within 7 days the lesion was appeared. 60-g rainbow trout for held at 10°C, the
50% lethal dose of Pd-10 was >107 CFU, compared with 8.1×105CFU/ml for the parent strain
(Leung et al., 1995). After 15 days infection produced 50% mortality with 2.7×104CFU/ml
(Table 1). Since 50% mortality was observed at 2.7×104CFU /ml concentration. This dose was
chosen for all the experiments.
To preserve and protect the environment as well as human health as a best alternative,
different parts of Azadirachta indica (Neem) tree have been studied by Chitmanat et al. (2005)
Indian almond (Terminalia catappa) and garlic (Allium sativum) have been said as an alternative
to chemicals to treat fish ectoparasites, Trichodina sp. infections in tilapia (O. niloticus)
fingerlings. Both Indian almond and garlic had low acute toxicity to tilapia fingerlings, treating
the Trichodiniasis caused by Trichodina. The immunostimulant effects of the dietary intake of 3
plants (viz., Viscum album, Urtica dioica and Zingiber officinale)-extracts on rainbow trout
(Oncorhynchus mykiss) have also been narrated by the authors. Christybapitaetal.(2007)
observed the immunostimulatory effect of aqueous extract (AqE) of Eclipta alba (Bhangra) leaf
(oral administration as feed supplement) in tilapia fish, Oreochromis mossambicus. It was noted
that the E. alba extract enhances non-specific immune responses and disease resistance of O.
9
mossambicus against A. hydrophila infection. According to Winkaler et al. (2007), A. indica

extract can be used successfully in aquaculture to control fish predators. Ravikumar et al. (2010)
observed that the chloroform extract of Datura metel plant has wide range of antimicrobial
activity against many fish pathogens. D. metel which collected from the Kanyakumari coast can
be used as a putative antimicrobial drug in the aquaculture maintenance. The chloroform extract
of D. metel can be effectively used as a potential antimicrobial agent to overcome the problem of
mass mortality of ornamental fish in aquarium so as to enable to enhance the market revenue
throughout the world. These authors also told the antimicrobial activity of 5 Chinese herb
extracts against 13 bacterial and 2 viral fish pathogens. Sharma et al. (2010) observed the
stimulatory effect of dietary doses of Withania somnifera (Ashwagandha) root on immunity and
disease resistance against A. hydrophila infection in Indian major carp, L. rohita fingerlings.
Abdul Kader Mydeen and Haniffa (2011) cited that A. indica leaf AqE could effectively control
the A. hydrophila infection in common carp (a fresh-water fish usually bred in ponds), Cyprinus
carpio. Further, Enterobacter sp. and Escherichia coli bacteria, isolated from marine fish
(Amphiprion sebae) showed 15 mm zone of inhibition against neem extract. The herb-
supplements/additives included in the fish feeds usually maintain and improve the physiological
functions. Ahilanet al. (2010) reported that the herbs have significant role in aquaculture. The
herbal growth promoters in the carp fish feeds showed beneficial effects. There was a significant
difference between different herbal additives on the effect of growth rate in goldfish. The
synergistic effects of herbs have been found in Clarias gariepinus (Turan and Akyurt, 2005) and
Japanese flounder (Ji et al., 2007).
Conclusion:
Medicinal plants are important elements of traditional medicine in the virtually all
cultures and promise a cheaper source for therapeutics. Herb powder added fish feed are cheap
and having lesion healing potency. The present study revealed that V. zizanioides traditional
medicinal plant possesses potential antimicrobial activity against fish pathogenic A. hydrophila
with A. invadans.
Reference:
Abdul Kader Mydeen K.P. and Haniffa M.A. (2011). Evaluation of antibacterial activity
ofmedicinal plants on fish pathogen,Aeromonas hydrophila. Journal ofResearch Biology.
1: 1-5.
10
(ISBN: 978-93-91768-45-4)
Ahilan, B., Nithiyapriyatharshini, A., and Ravaneshwaran, K., (2010). Influence of certain herbal
additives on the growth, survival and disease resistance of goldfish, Carassius auratus
(Linnaeus). Tamilnadu J. Vet. Ani. Sci., 6(1): 5-11.
Balasubramanian, V. (2005). Physiology of Cyprinus carpio infected by Aeromonas hydrophila
and efficacy of a selected neem formulation for the treatment of infected fish. Ph. D.,
Thesis, Madurai Kamaraj University, Madurai, India.
Chinabut, S., Roberts, R.J. Willoughby, L.G., and Pearson, M.D. (1995). Histopathology of
snakehead, Channastriatus (Bloch), ex-perimentally infected with specific Aphanomyces
fungus associated with epizootic ulcerative syndrome (EUS) at different tempera-tures,
Journal of Fish Diseases 20: 135-144.
Chitmanat, C., Tongdonmuan, K., and Nunsong, W. (2005). The use of crude extracts from
traditional medicinal plants to eliminate Trichodina sp. in tilapia (Oreochromis niloticus)
fingerlings. Songklanakarin J. Sci. Technol., 27(Suppl. 1): 359-364.
Christybapita, D., Divyagnaneswari, M., and Michael, R.D., (2007). Oral administration of
Eclipta alba leaf aqueous extract enhances the non-specific immune responses and disease
resistance of Oreochromis mossambicus. Fish Shellfish Immunol., 23(4): 840-852.
Das, M.K. (1997). Epizootic Ulcerative Syndrome (EUS) in fishes - its present status in India.
CICFRI (ICAR), Bull. No. 69.
Ji, S.C., Takaoka, O., Jeong, G.S., Lee, S.W., Ishimaru, K., Seoka, M., and Takii, K., (2007).
Dietary medicinal herbs improve growth and some non-specific immunity of red seabream
Pagrus major. Fish. Sci., 73: 63-69.
Joseph, S.W. and Camahan, A. (1994). The isolation, identification and systematic of the motile
Aeromonas species. Annu. Rev. Fish Dis. 4: 315-343.
Leung, K.Y., Low, K.W., Lam, T.J. and Sin. Y.M. (1995). Interaction of the fish pathogen
Aeromonas hydrophila with tilapia, Oreochromis aureus (Steindachner), phagocytes. J.
Fish Dis.18: 435– 447.
Muthukrishnan, D., Mohamed, A.K.H., Chidambaram, M.R.S., and Sing., V.A., (2008).
Microbial flora from the Epizootic Ul-creative Syndrome (EUS) infected
murrelChannastriatus (Bloch, 1797) in Tirunelveli Region. Turkish Journal of Veterinary
and Animal Sciences. 32(3): 221-224.
Ravikumar, S., Palani Selvan, G., and AnithaAnandhaGracelin, N. (2010). Antimicrobial activity
of medicinal plants along Kanyakumari coast, Tamil Nadu, India. Afr. J. Basic Appl. Sci.,
2(5-6): 153-157.
Scharperculeus, W. (1991). Text book of Fish diseases.1: 75-102.
11
Shariff, M. and Subasinghe, R. (1994). Experimental induction of epizootic ulcerative syndrome

e(EUS). ODA Reginal Seminar on epizootic ulcerative syndrome. In: Roberts, R.J.,
Campbell, B. and MacRae, I.H (Eds.). The Aquatic Animal Health Research Institute,
Bangkok. 273: 25-27.
Sharma, A., Deo, A.D., Riteshkumar, S.T., Chanu, T. I., and Das, A. (2010). Effect of Withania
somnifera (L. Dunal) root as a feed additive on immunological parameters and disease
resistance to Aeromonas hydrophila in Labeo rohita (Hamilton) fingerlings. Fish Shellfish
Immunol., 29(3): 508-512.
Sprague, J.B. (1973). The ABC‘s of pollutant bioassay using fish. Biological methods for the
assessment of water quality. ASTM STP 528. American Society for Testing and Materials,
Philadelphia. 6-30.
Stevens, R.B. 1974. Mycology Guidebook. University of Washington Press, Seattle. National
Science Foundation to the Mycological Society of America. 703 pp.
Supap, P. (1985). Histopathological and hematological studies of walking catfish (Clarias
batrachus Linn.) infected by Aeromonas hydrophila. M.S. Thesis (Fisheries Science),
Kasetsart University, Bangkok.
Turan, F., and Akyurt, I., (2005). Effects of red clover extract on growth performance and body
composition of African catfish, Clarias griepinus. Fish. Sci., 71: 618- 620.
Vaidyarathnam, P.S.V. (1995). Indian medicinal plants, Aryavaidyasala, kottakkal, orient
Longman Ltd. Chennai.
Vijayakumari, P., and Sukumaran, N.P. (2014). Effect of Euphorbia hirta plant leaf extract on
immunostimulant response of Aeromonas hydrophila infected Cyprinus carpio. Peer
journal, 2: 1-17.
Winkaler, E. U., Santos, T.R.M., Machado-Neto, J.G. and Martinez, C.B.R. (2007). Acute
lethal and sub-lethal effects of neem leaf extract on the neotropical freshwater fish,
Prochiloduslineatus. Comp. Biochem.Physiol. Part C, 145: 236-244.
Yambot, A.V. (1998). Isolation of Aeromonas hydrophila from Oreochromis niloticus during
fish disease outbreaks in the Philippines. Asian Fisheries Society, Manila,
Philippines.10:347-354.
12
(ISBN: 978-93-91768-45-4)
POTENTIAL OF WHEATGRASS BASED FORMULATED FEED IN FRESH

WATER FISH CULTURE
S. S. Butle1, R. A. Gulhane1 and P. S. Joshi2
1Department of Zoology,
S. S. S. K. R. Innani Mahavidyalaya, Karanja (Lad), Dist. Washim, Maharashtra, India
Shri Shivaji Arts, Commerce and Science College, Akot, Dist. Akola, Maharashtra, India
Corresponding author E-mail: [email protected],
[email protected], [email protected]
Background:
The severe world population explosion is presenting complex challenges, being the most
important one is malnutrition and shortage of food in terms of quantity and quality. Milk, meat
and eggs are the important animal source of protein. Animal production is a long term project to
produce adequate amount of quality protein to meet the national demand. Meat production in
terms of demanding protein requirement is improving with the establishment of poultry industry
and fish farming. The production of quality protein is associated with the development of
fisheries on commercial basis. Fish production sector is very important not only as a main source
of animal protein to ensure food security but also to improve employment and income for
poverty elimination in developing countries like India (Bhosale et al., 2010).
Fish is very important and common dietary animal protein source in human nutrition in
Indian subcontinent. Production of aquatic species through freshwater fisheries and aquaculture
for protein supply is being encouraged in developed or developing countries but in under-
developed countries, it is declining. According to nutritionists, fish is an excellent substitute for
red meat and an excellent source of protein. Fish flesh contains all the essential amino acid and
minerals like iodine, phosphorus, potassium, iron, copper and vitamin A and D in desirable
concentrations. It serves as valuable ingredient to a healthy diet because of its low carbohydrate
and unsaturated fat contents. It is often recommended by doctors to heart patients since it is an
excellent source of Omega 3. So the inclusion of fish in our diet can make a valuable
contribution to any diet that contain mainly of cereals, starchy roots and sugar for the healthy
growth (Corto-Martinez et al., 2007)
13
The fresh water fish is highly demanded as a food in India and adjacent countries. To
satisfy the increasing demand of fish as food, it is important to adopt modern practices of
aquaculture. As part of these practices, supplementation of nutrients to the fish is very necessary.
Good nutrition in animal production systems is essential to economically produce a healthy, high
quality product. In fish farming, nutrition is critical because feed represents 40-50% of the
production costs. Fish nutrition has advanced dramatically in recent years with the development
of new, balanced commercial diets that promote optimal fish growth and health. The
development of new species-specific diet formulations supports the aquaculture industry as it
expands to satisfy increasing demand for affordable, safe, and high-quality fish products (Li et
al., 2013).
Now a day, varieties of fish meals are easily available in market. Fishmeal is a
commercial product mostly made from fish that are not generally used for human consumption; a
small portion is made from the bones and left over from processing of fish used for human
consumption, while the larger percentage is manufactured from wild-caught, small marine fish;
either unmanaged by-catch or sometimes sustainable fish stocks. It is powder or cake obtained
by drying the fish or fish trimmings, often after cooking, and then grinding it. If the fish used is
a fatty fish it is first pressed to extract most of the fish oil. Fish meal production is a significant
contributor of over-fishing, and risks pushing fisheries beyond their replacement rate (Sahu et
al., (2007).
The use of feedadditives is beneficial in increasing fish production. The feedadditives are
also the supplements for farm animals that cannot get enough nutrients from regular meals that
the farmers provide and it includes vitamins, amino acids, fatty acids, and minerals. In some
cases, if an animal does not have some specific nutrition in its diet then it may not grow properly.
The nutritional values of animal feeds are influenced not only by their nutrient content, but also
by many other factors. These include the feed presentation, hygiene, digestibility, and effect on
intestinal health. Even with all of the benefits of higher quality feed, most of a farm animal's diet
still consists of maize, wheat and soybean meal because of the higher costs of quality feed.
Hence feed and feeding are among the most important factors influencing growth, feed
utilization and tissue composition of the fish in intensive culture (Okumus and Mazlum, 2002).
In intensive culture systems fish production per unit of surface can be increased at well,
as long as sufficient oxygen, fresh water and food are provided. Because of the requirement of
sufficient fresh water, a massive water purification system must be integrated in the fish farm.
One way to achieve this is to combine hydroponic horticulture and water treatment, see below.
The exception to this rule is cages which are placed in a river, which supplements the fish crop
14
(ISBN: 978-93-91768-45-4)
with sufficient oxygenated water. Some environmentalists object to this practice (Aly et al.,
2008).
The cost of inputs per unit of fish weight is higher than in extensive farming, especially
because of the high cost of fish feed. It must contain a much higher level of protein. These higher
protein-level requirements are a consequence of the higher feed efficiency of aquatic animals.
Fish such as salmon have Fish do not use energy to keep warm, eliminating some carbohydrates
and fats in the diet, required to provide this energy. This may be offset, though, by the lower land
costs and the higher production which can be obtained due to the high level of input control.
Aeration of the water is essential, as fish need a sufficient oxygen level for growth. This is
achieved by bubbling, cascade flow, or aqueous oxygen (Butle et al., 2018).
In fish culture, the risk of infections by parasites such as fish lice, intestinal worms,
bacteria, and protozoa is similar to that in animal husbandry, especially at high population
densities. However, animal husbandry is a larger and more technologically mature area of human
agriculture and has developed better solutions to pathogen problems. Intensive aquaculture has to
provide adequate water quality levels to minimize stress on the fish. This requirement makes
control of the pathogen problem more difficult. Intensive aquaculture requires tight monitoring
and a high level of expertise of the fish farmer (Metwally, 2009 a, b).
The formulated diet can also prove to be effectives in increasing fish production. The
main issue in formulating feed is to meet the protein and essential amino acids requirements of
the species. Fishmeal is generally the preferred protein source because of the high quality of the
protein and its essential amino acids profile. However, fishmeal is generally expensive and is not
always available. It is economically judicious to replace fishmeal with alternative protein sources
including animal by-products, oilseed meal and cakes, legumes and cereal by-products and
aquatic plants. Most of these ingredients are deficient in some essential amino acids and hence
require supplementation or be compensated with other feedstuffs. Although most of the oilseed
cakes or by-products are generally deficient in lysine and methionine, blending of different
oilseed cakes often provides balanced amino acid profile. However they contain many anti-
nutritional factors such as gossypol, glucosinolates, saponins, trypsin inhibitors etc. which limit
their use in compound feeds or require removal or inactivation through specific processing such
as heating, cooking etc. Hence, the formulation of diet is necessary with sophisticated method
help to preserve the nutrients for rapid growth of fish (Farahi et al., 2010).
The protein is the most expensive part of fish feed; hence it is important to accurately
determine the protein requirements for each species and size of cultured fish. Proteins are formed
15
by linkages of individual amino acids. Fish feeds prepared with plant like soybean meal protein
typically are low in methionine; therefore, extra methionine must be added to soybean-meal
based diets in order to promote optimal growth and health. Lipids are high-energy nutrients that
can be utilized to partially spare protein in aquaculture feeds. A recent trend in fish feeds is to
use higher levels of lipids in the diet. Although increasing dietary lipids can help reduce the high
costs of diets by partially sparing protein in the feed, problems such as excessive fat deposition
in the liver can decrease the health and market quality of fish (Diegane and Fall, 2011).
Carbohydrates are the most economical and inexpensive sources of energy for fish diets.
Although not essential, carbohydrates are included in aquaculture diets to reduce feed costs and
for their binding activity during feed manufacturing. Dietary starches are useful in the extrusion
manufacture of floating feeds. Cooking starch during the extrusion process makes it more
biologically available to fish. Vitamins are organic compounds necessary in the diet for normal
fish growth and health. They often are not synthesized by fish, and must be supplied in the diet.
Minerals are inorganic elements necessary in the diet for normal body functions. They can be
divided into two groups (macro-minerals and micro-minerals) based on the quantity required in
the diet and the amount present in fish. Common macro-minerals are sodium, chloride,
potassium and phosphorous. These minerals regulate osmotic balance and aid in bone formation
and integrity (Lee et al., 2012).
Micro-minerals (trace minerals) are required in small amounts as components in enzyme
and hormone systems. Common trace minerals are copper, chromium, iodine, zinc and selenium.
Fish can absorb many minerals directly from the water through their gills and skin, allowing
them to compensate to some extent for mineral deficiencies in their diet. Dietary nutrients are
essential for the construction of living tissues. They also are a source of stored energy for fish
digestion, absorption, growth, reproduction and the other life processes. The nutritional value of
a dietary ingredient is in part dependant on its ability to supply energy (Agatha, 2012).
Feeding rates and frequencies are in part a function of fish size. Small larval fish and fry
need to be fed a high protein diet frequently and usually in excess. Small fish have a high energy
demand and must eat nearly continuously and be fed almost hourly. Feeding small fish in excess
is not as much of a problem as overfeeding larger fish because small fish require only a small
amount of feed relative to the volume of water in the culture system. As fish grow, feeding rates
and frequencies should be lowered, and protein content reduced. However, rather than switching
to a lower protein diet, feeding less allows the grower to use the same feed (protein level)
throughout the grow-out period, thereby simplifying feed inventory and storage (Megbowon et
al., 2013).
16
(ISBN: 978-93-91768-45-4)
Feeding fish is labor-intensive and expensive. Feeding frequency is dependent on labor

availability, farm size, and the fish species and sizes grown. Large catfish farms with many
ponds usually feed only once per day because of time and labor limitations, while smaller farms
may feed twice per day. Generally, growth and feed conversion increase with feeding frequency.
In indoor, intensive fish culture systems, fish may be fed as many as 5 times per day in order to
maximize growth at optimum temperatures.
Many factors affect the feeding rates of fish. These include time of day, season, water
temperature, dissolved oxygen levels, and other water quality variables. For example, feeding
fish grown in ponds early in the morning when the lowest dissolved oxygen levels occur is not
advisable. In contrast, in recirculating aquaculture systems where oxygen is continuously
supplied, fish can be fed at nearly any time. During the winter and at low water temperatures,
feeding rates of warm-water fishes in ponds decline and feeding rates should decrease
proportionally. Feed acceptability, palatability and digestibility vary with the ingredients and
feed quality. Fish farmers pay careful attention to feeding activity in order to help determine feed
acceptance, calculate feed conversion ratios and feed efficiencies, monitor feed costs, and track
feed demand throughout the year. Published feeding rate tables are available for most commonly
cultured fish species. Farmers can calculate optimum feeding rates based on the average size in
length or weight and the number of fish in the tank, raceway, or pond (Lee et al., 2014).
The hormones, antibiotics and several other chemicals have been tested as growth
promoters, antibacterial and for other purposes in aquatic animals, but their use in aquatic animal
production cannot be recommended due to the residual effects in the muscle of fish as well as
prawns. Plants are natural sources of safer and cheaper chemicals. The beneficial effects of
bioactive plant substances in animal nutrition may include the stimulation of appetite and feed
intake, growth promotion, the improvement of endogenous digestive enzyme secretion, and
activation of immunostimulation and antibacterial, antiviral and antioxidant actions in
aquaculture practices. But in recent years, the concern about bacterial resistance to antibiotics in
livestock industry has led to legislation minimizing or eliminating the use of such compounds
(Lee et al., 2012).
Wheatgrass Triticum aestivum refers to young grass of the common wheat plant, which
belongs to Poaceae family. This is the most commonly found herb in India which is freshly
juiced or dried into powder for animal and human consumption. Wheatgrass‘ culms are simple,
hollow or pithy, glabrous, and the leaves are approximately 1.2 m tall, flat, narrow, 20-38 cm
long and 1.3 cm broad. The spikes are long, slender, dorsally compressed and somewhat
17
flattened (Mujoriya and Bodla, 2011; Sareen et al., 2014). The phytochemical and
pharmacological potential of wheat grass is well reviewed by Durairaj et al. (2014);
Suriyavathana et al. (2016). Wheatgrass is a source of potassium, dietary fiber, vitamin A,
vitamin C, vitamin E, vitamin K, thiamin, riboflavin, niacin, vitamin B6, pantothenic acid, iron,
zinc, copper, manganese and selenium. Wheatgrass is also a source of protein. Kumar et al.
(2016) quoted that wheat grass is a nature‘s wonderful medicine that helpful in treatment of
various diseases and infections. Plant has been shown to have anti-inflammatory, antioxidant,
anticarcinogenic, immunomodulatory, laxative, astringent, diuretic, antibacterial, antihemolytic
and anti-aging properties as well improve reproductive health. Its use in acidity, colitis, kidney
malfunctions, atherosclerosis and swelling has been shown to be beneficial (Sharma et al., 2016;
Johri and Khan, 2017, Uraiwan et al., 2019).
As well, this herb is useful for the animals in variety of ways. Aabdullahet al. (2014)
mentioned that the wheat flour and bran is common ingredient in animal feed. The wheat bran
mixed with fish meal improved the growth performance and meat quality in cattle. Emmanuelle
et al. (2014) quoted that wheat gluten can be used as an alternative to protein in feed for
carnivorous fish. Nath et al. (2014) perform the successful rearing of Asian catfish fry (Clarias
batrachus) with wheatgrass powder mixed formulated feed in plastic half drum. Islam et al.
(2017) observed that potential of wheatgrass based feed for stinging catfish fry nursing in
laboratory condition. Johri and Khan (2017) had given the phytochemical characterization of
wheat grass and observed antioxidant and anti-haemolytic potential. Gasco et al. (2018)
suggested that addition of easily available phytochemically effective herbs into the fish diets help
to improve the feed conversion efficiency or elevate general conditions for fish growth and
maintenance (Makode et al., 2018; Uraiwan et al., 2019).
Review of Earlier studies

The herbs and herbal products extract contains various bioactive compounds which
potential source of natural antioxidants with medicinal properties. Their addition to the feed
cures many diseases, promote growth, reduce stress, improve immunity and prevent infections in
fish under culture. According to Cristea et al. (2012), the supplementation of herbs and herbal
products in fish diet is cheaper and environmental friendly with no side effects to the fish and
consumers. Hence their use may lead to achieve an augmented aquaculture production inan eco-
friendly, cost effective and sustainablemanner is gaining popular. In recent years, many studies
are being carried to study the effectiveness of herbal supplementation in fish feed to manage fish
growth and produce healthy fish. The outcomes of these studies suggested that the herbal feed
18
(ISBN: 978-93-91768-45-4)
supplements promote growth, minimizes stress, improves immunity and prevents various
infections in fishes that will help to produce healthy fishes for humanconsumption (Shakya,
2017). The present review highlights the importance of few effective herbs and herbal products
supplementation in fish feed for better fish production.
Omoregie (2001) studied the utilization and nutrient digestibility of mango seeds
Mangifera indica and palm Kernel Elaeis guineensis meal by juvenile Labeo senegalenis.
Citarasu et al. (2002) developed Andrographis paniculata, Eclipta erecta, Hygrophila spinosa,
Ocimum basilium, Phyllanthus niruri, Picrorhiza kurooa, Psoralea corylifolia, Solanum
trilobatum, Tinospora cordifoliaare and Zingiber officinalis enriched herbal diet for producing
quality larvae in Penaeus monodon. According to Brannas et al. (2003) and Elkayam et al.
(2003), the use of such natural supplements in formulated diet, increased their behavioural and
growth performances. Skidmore-Roth (2003) mentioned that Acalypha indica, Aegle marmelos,
Chrysopogon zizanioides, Cymbopogon citratus, Hordeum vulgare, Medicago sativa, Senna
auriculata, Sorghum bicolor and Urtica dioica are some of the most useful medicinal herbs that
can be used as natural supplements. Shalaby et al. (2004) observed the response of Nile tilapia,
Oreochromis niloticus, fingerlings to diets supplemented with different levels of fenugreek
seeds.
Chitmanat et al. (2005) isolated antiparasitic, antibacterial, and antifungal solution from
Terminalia catappa against some tilapia Oreochromis niloticus pathogens. Micol et al. (2005)
observed that olive leaf extract exhibits antiviral activity against viral haemorrhagic Septicaemia
rhabdo virus. Turan (2006) improved the growth performance in tilapia Oreochromis aureus by
supplementation of red clover Trifolium pratense in diets. Yin et al. (2006) noticed the
effectively of two Chinese herbs Astragalus radix and Scutellaria radix on non-specific immune
response of tilapia Oreochromis niloticus (Zakes et al., 2008). John et al. (2007) found the
effectiveness of Eichinacea purpurea, Nigella sativa and Origanum marjoranaas as feed
additives on the survival, growth performance and immune response of Nile tilapia Oreochromis
niloticus.
Ardo et al. (2008) noticed that Chinese herbs Astragalus membranaceus and Lonicera
japonica with boron enhance the non-specific immune response of Nile tilapia Oreochromis
niloticus and resistance against Aeromonas hydrophila. Ashraf and Goda (2008) mentioned that
dietary Ginseng herb Eleutherococcus senticosus supplementation improved growth, feed
utilization, and hematological indices of Nile Tilapia, Oreochromis niloticus fingerlings.
19
Pachanawan et al. (2008) observed the potential of Psidium guajava supplemented fish diets in
controlling Aeromonas hydrophila infection in tilapia Oreochromis niloticus.
Thy et al. (2008) studied the effect of water spinach Spinacia oleracea and duckweed
Lemna on fish growth performance in poly-culture ponds. Won et al. (2008) found that residum
extract of Siberian ginseng Eleutherococcus senticosus increased non-specific immunity in olive
flounder Paralichthys olivaceu. Yuan et al. (2008) found the appreciable effects of Astragalus
membranaceus extracts on the expression of immuneresponse genes in head kidney, gill and
spleen of the common carp, Cyprinus carpio. Zakes et al. (2008) studied the effects of Lonicera
japonica on the growth performance and body composition of juvenile pike perch (Sander
lucioperca).
Ergun et al., (2009) proved the influence of Ulva lactuca meal on growth, feed
utilization, and body composition of juvenile Nile tilapia Oreochromis niloticus at two levels of
dietary lipid. Jeney et al. (2009) mentioned that addition of different single herbal extracts of the
herbs Artemisia capillaries, Cnidium officinale, Crataegi fructus, Glycyrrhiza glabra, Isatis
tinctoria, Massa medicate and Polygonum multiflorum promoted the growth and enhanced some
non-specific immunity indicators of fish. Omoregie et al., (2009) studied effect of varying levels
of sweet potato Ipomea batatas peels on growth, feed utilization and some biochemical
responses of the Cichlid Oreochromis niloticus (Faramarzi et al., 2012).
Citarasu (2010) noticed that Ricebran Oryza sativa improved the survival and
reproductive performance in fish Artemia parthenogenetica. Sharma et al. (2010) observed the
efficacy of Withania somnifera root as a feed additive on immunological parameters and disease
resistance to Aeromonas hydrophila in Labeo rohita fingerlings. Ahmad and Abdel-Tawwab
(2011) used the caraway Carum carvi seed meal as a feed additive in fish diets to improve
growth performance, feed utilization, and whole-body composition of Nile tilapia Oreochromis
niloticus. Chakraborty and Hancz (2011) studied the application of phytochemicals as
immunostimulant, antipathogenic and antistress agents in finfish culture.
Coutteau et al. (2011) mentioned that extracts of Allium tuberosum, Aniba rosaeodora,
Capsicum annum longum, Cinnamomum zeylanicum, Elettaria caramomum, Mentha piperita,
Myristica flagrans, Piper nigrum, Salvia apiana and Syzygium aromaticum improves the
productivity and economics in aquaculture.Joseph et al. (2011) observed the influence of
Crossandra infundibuliformiss, Hibiscus rosasinensis, Ixora coccinea and Rosa indica flowers
on the growth and colouration of orange sword tail Chicilidae fish Xiphophorus hellerei.
Kaleeswaran et al. (2011) observed the growth response, feed conversion ratio and antiprotease
activity in Catla catla fed on Cynodon dactylon mixed diet. Obaroh and Achionye-Nzeh (2011)
20
(ISBN: 978-93-91768-45-4)
studied the effects of crude extract of Azadirachtaindica leaves at controlling prolific breeding
in Oreochromis niloticus.
Arumugam et al. (2012) observed the effect of dietary Nelumbo nucifera in growth and
haematology of Cirrhinus mrigala challenged with Pseudomonas aeruginosa. Benny et al.
(2012) investigated the immunostimulatory behaviour of Musa acuminate peel extract in
Clarias batrachus. Falaye et al. (2012) replaced the maize Zea mays using cowpea Vigna
unguiculata hull meal in practical feeds of African catfish Clarias gariepinus. Jha et al. (2012)
studied the effects of marigold flower Beta vulgaris and beetroot Calendula officinalis meals on
growth performance, carcass composition, and total carotenoids of snow trout Schizothorax
richardsonii. Sivagurunathan et al. (2012) studied the immunostimulatory potential of dietary
amla Phyllanthus emblica in growth and hematology of Tilapia mossambicus challenged with
Pseudomonas aeruginosa. Yilmaz et al. (2012) observed the effects of herbal supplements on
growth performance, change in body composition and some blood parameters of sea bass
Dicentrarchus labrax.
Hwang et al. (2013) observed that dietary green tea extract improves the growth
performance, body composition, and stress recovery in juvenile black rockfish, Sebastes
schlegeli. Mamman et al. (2013) studied the hematological indices of Clarias griepinus
fingerlings fed diet containing graded levels of calabash Lagenaria vulgaries seedmeal. Borkar
et al. (2014) evaluated the impact of shatavari Asparagus racemosus and ashwagandha
Withanias ominiferaon average body weight of freshwater fish Channa punctatus. Gaber et al.,
(2014) observed the mentioned that dietary Alterna storini, Hippophaer hamnoides Phoenix
dactylifera and Thymus vulgaris improves the growth performance in Nile tilapia Oreochromis
niloticus fingerlings. Karpagam and Krishnaveni (2014) studied the effect of Supplementation
of Moringa oleifera, Ocimum sanctum, Sesbania grandiflora and Solanum verbascifolium
leaves as Growth Promoters of Tilapia Fish Oreochromis mossambicus.
Adel et al. (2015) observed the effects of dietary peppermint Mentha piperita on growth
performance, chemical body composition and hematological and immune parameters of fry
Caspian white fish. Labh and Shakya (2016) studied the effects of dietary Choerospondias
axillaris on survival, growth and protein profile of common carp Cyprinus carpio fingerlings.
Kaur and Shah (2017) mentioned the efficacy of Vegetable Colour from Red Sandal
Pterocarpus santalinus on acceptability, colour development and growth of Tilapia Tilapia
mossambica. Makode (2017) studied the effects of dietary onion on growth performance in the
fresh water fish Clarias batrachus.
21
Butle et al. (2018) studied the dietary garlic Allium sativum induced effects on behaviour
responses, growth performance and feed utilization in Clarias batrachus. Plaza et al. (2018)
observed the effect ofspirulina Arthrospira platensis supplementation on tilapia Oreochromis
niloticus growth and stress responsiveness under hypoxia. Rodge et al. (2018) observed the
effects of dietary garlic Allium sativum on hematology and biochemistry of Clarias batrachus.
Gabriel et al., (2019) studied the effect of dietary Aloe vera on growth performance, feed
utilization, hemato-biochemical parameters, and survival at low pH in African catfish Clarias
gariepinus. Roghieh et al., (2019) assessed the effects of Coriandrum sativum as feed additive
on mucosal immune parameters, antioxidant defense and, immune‐related genes expression in
Zebrafish Danio rerio. Like all these above herbs, wheatgrass is well known for its medicinal
properties. Butle et al. (2019) evaluated the phytochemical and antioxidant potential of aqueous
wheatgrass Triticum aestivum extract.
Abdus et al. (2020) conducted an experiment to produce hydroponic wheatgrass and
feeding trials with stinging catfish, rohu and grass carp. Abdus et al. (2020) observed the growth
response of juvenile rohu (Labeo rohita) to wheatgrass powder supplemented diet. They
observed the effects of wheat sprout improved the growth, survival and production of
experimental fish. Rana et al. (2020) studied the dietary supplementation of wheatgrass powder
to assess somatic response of juvenile grass carp (Ctenopharyngodon idella).The overall somatic
performance of grass carp fed wheatgrass supplemented test diets was satisfactory compared to
control diet without wheatgrass. Notably, fish survival was substantially improved. The feed
conversion ratio value and feed formulation cost of the respective diets were reduced in a good
amount.
The phytochemical screening of the aqueous extract of wheatgrass showed the presence
of various secondary metabolites. As well wheatgrass was proved to be an effective in different
antioxidant assays. These potential varieties of wheatgrass suggested the effective utility in fresh
water fish culture as dietary supplementation.It will beprove to be beneficial for the successful
improvement of fisheries by achieving maximum yields with using safer and cheaper formulated
feed to induce developmental performance of fresh water fishes.
References:
Abdullah A., Osman M., Isam K., Waleed A., Goddard S. (2014). Effect of feeding fish-wheat
bran mean on performance and meat quality of Omani Sheep. J. Appl. Ani. Res., 45:1-4.
Abdus S. M., Das S. C., Noor A. M., Sultana M. (2020). Development of hydroponic
wheatgrass cultivation as supplementary feed production techniques for fish and poultry
22
(ISBN: 978-93-91768-45-4)
farming in Bangladesh. Poster Published by Department of Aquaculture, Bangladesh

Agriculture University, Mymensingh.
Abdus S., Shakil R., Rakib A., Noor M. (2020). Growth response of juvenile rohu (Labeo
rohita) to wheatgrass powder supplemented diet. Res. Agri. Live.Fish. 7(3): 533-543
Adel M., Abedian-Amiri A., Zorriehzahra J., Nematolahi A., Esteban M. (2015). Effects of
dietary peppermint (Mentha piperita) on growth performance, chemical body composition
and hematological and immune parameters of fry Caspian white fish (Rutilusfrisii-
kutum).Fish Shellfish Immunol. 45 (2):841-7.
Agatha A. N. (2012). The Effect of Garlic (Allium sativum) on Growth and Haematological
Parameters of Clarias griepinus(Burchell, 1822) Sustain. Agr. Res. 1 (2): 222-228
Ahmad M. H., Abdel-Tawwab M. (2011). The use of caraway Carum carvi seed meal as a feed
additive in fish diets: Growth performance, feed utilization, and whole-body composition
of Nile tilapia, (Oreochromis niloticus ) (L.) fingerlings. Aquacult.31(4):110-114.
Aly S. M., Nashwa M. A., Mohamed M. F. (2008). Effect of garlic on the survival, growth,
resistance and quality of Oreochromis niloticus .8th Int. Sympo. Tilapia Aqua.: 277-295.
Ardo L., Yin G., Xu P., Varadi L., Szigeti G., Jeney Z. (2008). Chinese herbs
(Astragalusmembranaceusand Lonicera japonica) and boron enhance the non-specific
immune response of Nile tilapia (Oreochromis niloticus ) and resistance against
Aeromonas hydrophila. Aquaculture.275:26-33.
Arumugam S., Innocent B., Muthulakshmi S. (2012).Immunomodulatory effect of dietary
Nelum bonucifera (lotus) in growth and haematology of Cirrhinus mrigala challenged
with Pseudomonas aeruginosa. J Appl Pharm Sci. 2:191-195.
Ashraf M. A., Goda, S. (2008).Effect of dietary Ginseng herb (Ginsana® G115) supplementation
on growth, feed utilization, and hematological indices of Nile Tilapia, Oreochromis
niloticus (L.), fingerlings.J World Aquac Soc. 39(2):205–214.
Benny P., Viswanathan G., Thomas S., Nair A. (2012).Investigation of
immunostimulatorybehaviour ofMusa acuminate peel extract in Clarias batrachus. IIOAB
Journal. 1(2):39-43.
Bhosale S. V., Bhilave M. P., Nadaf S. B. (2010). Formulation of Fish Feed using Ingredients
from Plants.Res. J. Agric. Sci. 1:284-287.
Bloch M. E. (1793).Naturgeschichte der ausländischenFische.Mitsechs und
dreissigausgemaltenKupfernnachOriginalen.SiebenterTheil. - pp. I-IX, 1-144. Berlin.
Borkar S. B., Rathod S. H., Kulkarni K. M., Tantarpale V. T. (2014).Impact of shatavari and
ashwagandha on average body weight of freshwater fish Channa punctatus.Journal of
Global Biosciences. 3(2): 582-585.
23
Brannas E., Jonsson S., Lundquist H. (2003). Influence of food abundance on individual
behavior strategy and growth rate in juvenile brown trout (Salmo trutta). Can J Zool., 8(1):
684-691.
Butle S. S., Gulhane R. G., Joshi P. S. (2019). Evaluation of Phytochemical and Antioxidant
Potential of Aqueous Wheatgrass Extract for fish feed formulation, VIIRJ, 9(2):65-70.
Butle S. S., Thakare V. G., Joshi P. S. (2018).Study of dietary garlic induced effects on
behaviour responses, growth pśerformance and feed utilization in Clarias batrachus
(Linnaeus, 1758). Bioscience Discovery, 9(3): 403-408.
Chakraborty S. B., Hancz C. (2011).Application of phytochemicals as immunostimulant,
antipathogenic and antistress agents in finfish culture.Reviews in Aquaculture,3(3):103-
119.
Chitmanat C., Tongdonmuan K., Khanom P., Pachontis P., Nunsong W. (2005).Antiparasitic,
antibacterial, and antifungal activities derived from a terminaliacatappa solution against
some tilapia (Oreochromis niloticus ) pathogens. Actahorticulturae, 05:678-681.
Citarasu T. (2010). Herbal biomedicines: a new opportunity for aquaculture industry. Aquacult
Int. 18:403–414.
Citarasu T., Sekar R. R., Babu M. M., Marian M. P. (2002). Developing Artemia enriched
herbal diet for producing quality larvae in Penaeus monodon (Fabricius). Asian Fish
Sci.15:21-32.
Corto-Martinez, M., Corzo, N., Villamiel M. (2007).Biological properties of onions and garlic,
Trends in food Sci. Tech, 18; 609-625.
Coutteau P., Ceulemans S., Alexander H. (2011). Botanical extracts improve productivity and
economics in aquaculture. Nutriad Int.,Belgium. 11: 1-8
Cristea V., Antache A., Grecu I., Docan A. (2012).The use of phytobiotics in
aquaculture.University of Agricultural Sciences and Veterinary Medicine.IASI, 12:57-60.
Diegane N., Fall J. (2011). The effects of the garlic (Allium sativum) on growth and immune
responses of hybrid tilapia Oriochromis niloticus x Oriochromis aureus. J. Clin.
Immune.Immunepatho. Res. 3(1): 1-9.
Durairaj V, Hoda M, Shakya G, Pajaniradje S, Babu P and R Rajagopalan (2014).Phytochemical
screening and analysis of antioxidant properties of aqueous extract of wheatgrass. Asian.
Pac. J. Trop. Med.,7(1): 398-404.
Elkayam A., Mirelman D., Peleg E., Wilchek M., Miron T., Rabinkov A., Oron-Herman M.,
Rosenthal T. (2003).The effects of allicin on weight in fructose-induced hyperinsulinemic,
hyperlipidemic, hypertensive rats.Ameri. J. Hyper., 16(12): 1053-1056
Emmanuelle A, Aurélien F, Anne A and F Respondek (2013).Use of vital wheat gluten in
aquaculture feeds. Aqua.Bios.9 (21): 1-13.
24
(ISBN: 978-93-91768-45-4)
Ergun S., Soyuturk M., Guroy B., Guroy D., Merrifield D. (2009). Influence of Ulva meal on
growth, feed utilization, and body composition of juvenile Nile tilapia (Oreochromis
niloticus ) at two levels of dietary lipid. Aquacult Int. 17: 355-361.
Falaye A. E., Omoike A., Olasebikan O. B. (2012). Replacement of maize (Zea mays) using
cowpea (Vignaunguiculata) hull meal in practical feeds of African catfish (Clarias
griepinus) fry. International Journal of Plant, Animal and Environmental
Sciences.2(3):178-182.
Farahi, A., Kasiri M., Sudagar M., Iraei M., Shahkolaei M. (2010).Effect of garlic (Allium
sativum) on growth factors, some hematological parameters and body compositions in
rainbow trout (Oncorhynchus mykiss).AACL Bioflux, 3 (4): 317-323.
Faramarzi M.,Lashkarboloki M., Kiaalvandi S., Iranshahi F. (2012).Influences of different levels
of sweet potato peels on growth and feeding parameters and biochemical responses of
Cyprinus carpio(Cyprinidae).American- Eurasian J Agric. & Environ. Sci. 12(4):449-455.
Gaber M. M., Labib E. H., Omar E. A., Zaki M. A., Nour A. M. (2014). Effect of partially
replacing corn meal by wet date on growth performance in nile tilapia (Oreochromis
niloticus) fingerlings, diets supplemented with digestarom. Journal of Geoscience and
Environment Protection.2:60-67.
Gabriel N. N., Wilhelm M. R., Shimoosshil K. (2019). Effect of dietary Aloe
vera polysaccharides supplementation on growth performance, feed utilization, hemato-
biochemical parameters, and survival at low pH in African catfish (Clarias griepinus)
fingerlings.Int. Aquatic Research, 11: 57–72(2019)
Gasco L., Gai F., Maricchiolo G., Genovese L., Ragonese S., Bottari T., Caruso G.
(2018).Fishmeal alternativeprotein sources for aquaculture feeds. In Feeds for the
Aquaculture Sector; Springer: Berlin/Heidelberg, Germany, pp. 1–28.
Hwang J. H., Lee S. W., Rha S. J., Yoon H. S., Park E. S., Han K. H., Kim S. J. (2013). Dietary
green tea extract improves growth performance, body composition, and stress recovery in
the juvenile black rockfish, Sebastesschlegeli. AquacultInternat.21:525-538.
Islam T., Rana K. M., Salam M. A. (2017). Potential of wheatgrass based feed for stinging
catfish fry nursing in laboratory condition. International Journal of Fisheries and Aquatic
Studies, 5(6): 179-184.
Jeney G., Yin G., Ardo L., Jeney Z. (2009).The useof immunostimulating herbs in fish.An
overview of research.Fish Physiology and Biochemistry 35:669-676.
Jha G. N., Sarma D., Qureshi T. A., Akhtar M. S. (2012).Effect of marigold flower and beetroot
meals on growth performance, carcass composition, and total carotenoids of snow trout
(Schizothoraxrichardsonii).The Israeli Journal of Aquaculture.12:1-5.
25
John G., Mesalhy S., Rezk M., El-Naggar G., Fathi M. (2008). Effect of some
immunostimulants as feed additives on the survival and growth performance of Nile
tilapia, Oreochromis niloticus and their response to artificial infection. Egypt J AquatBiol
Fish.11(3):1299-1308.
Johri S., Khan N. (2017).Invitro Antioxidant and Antihaemolytic Potential of Triticumaestivum
Grass. Int. J. Compl. And Alt. Med. 9(5): 1-5.
Joseph B., Sujath S., Shalin J., Palavesam A. (2011). Influence of Four ornamental flowers on
the growth and colouration of orange sword tail Chicilidae fish (Xiphophorushellerei,
Heckel, 1940). International J. of Biological and Medical Research.2(3):621-626.
Kaleeswaran B., Ilavenil S., Ravikumar S. (2011).Growth Response, Feed Conversion Ratio and
Antiprotease Activity of Cynodondactylon(L.)Mixed Diet in Catla catla(Ham.).J Anim
Vet Adv. 10(4):511-517.
Karpagam B., Krishnaveni N. (2014).Effect of Supplementation of Selected Plant Leaves as
Growth Promoters of Tilapia Fish (Oreochromismossambicus).Res J Recent Sci. 3:120-
123.
Kaur R., Shah T. (2017).A review on role of plant waste products on fish growth, health and
production.Journal of Entomology and Zoology Studies 5(3): 583-589.
Kumar N, Murali M, Nair A and A Nair (2016).Green Blood Therapy of Wheat Grass - Nature‘s
Finest Medicine‘- A Literature Review.OSR-JPBS 11(2): 57-64.
Labh S. N., Shakya S. R. (2016). Effects of dietary lapsi (ChoerospondiasaxillarisRoxb.) on
survival, growth and protein profile of common carp (Cyprinuscarpio L) fingerlings. Int.
J. Zool. Stud. 1(5):36-415.
Lee D .H., Ra C. S., Song Y. H., Sung K. I., Kim J. D. (2012). Effect of dietary garlic extracts on
growth, feed utilization and whole body composition of juvenile sterlet sturgeon
(Acipenserruthenus). Asian Austra.. J. Anim. Sci. 25:577-583.
Lee D. H., Ra C. S., Song Y. H., Sung K. I., Kim J. D. (2014). Effects of dietary garlic powder
on growth, feed utilization, and whole body composition changes in fingerling starlet
sturgeon, Acipenserruthenus. Asian Austra.. J. Anim. Sci. 27:1303-1310.
Li M.,1Kaviarasu M., Zulkafli A. Subha B. (2013). Characterisation of Asian Snakehead
MurrelChannastriata(Channidae) in Malaysia: An Insight into Molecular Data and
Morphological Approach. The ScientificWorld Journal, 917506:1-16
Makode P. M. (2017). Effects of dietary onion on growth performance in the fresh water fish
Clarias batrachus (Linn.). Bioscience Discovery, 8(2):241-243.
Makode P., Pandharikar S., Joshi P., Gulhane R. (2018).Effects of dietary onion on behaviour of
the fresh water fish Channapunctatus (Bloch, 1793). ICRTST: 70-72.
26
(ISBN: 978-93-91768-45-4)
Mamman T., Ipinjolu J., Magawata I. (2013).Hematological indices of Clarias

griepinus(Burchell, 1882) fingerlings fed diet containing graded levels of calabash
(Lagenariavulgaries)seedmeal.JournalofBiology,Agricultureand Healthcare.3:100-104.
Megbowon, I., Adejonwo O., Adeyemi Y., Kolade O., Adetoye A., Edah B., Okunade O.,
Adedeji A. (2013).Effect of garlic on growth performance, nutrient utilization and survival
of an ecotype Cichlid.IOSR-JAVS, 6(3): 10-13.
Metwally M. A. (2009).Effect of Garlic (Allium sativum) on Some Heavy Metal (Copper and
Zinc) Induced Alteration in Serum Lipid Profile of Oreochromis niloticus World J. of Fish
and Mari. Sci. 1 (1): 01-06.
Metwally M. A. (2009). Effects of garlic (Allium sativum) on some antioxidant activities in
Oreochromis niloticus .World J. of Fish and Mari. Sci. 1 (1):56-64.
Micol V., Caturla N., Perez-Fons L., Mas V., Perez L., Estepa A. (2005). The olive leaf extract
exhibits antiviral activity against viral haemorrhagicsepticaemiarhabdovirus (VHSV).
Antiviral Research.66(2-3):129-136.
Mujoriya R., Bodla R. (2011). A study on wheat grass and its Nutritional value.Food Sci. Qual.
Mng. 2: 1-10.
Nath T., Hashem S., Salam M. (2014).Asian catfish fry (Clarias batrachus) rearing with
wheatgrass powder mixed formulated feed in plastic half drum. International Journal of
Fisheries and Aquatic Studies, 1(5): 162-168.
Obaroh I. O., Achionye-Nzeh G.C. (2011). Effects of Crude Extract of Azadirachtaindica Leaves
at Controlling Prolific Breeding inOreochromis niloticus (Linnaeus, 1758). Asian Journal
of Agricultural Research, 5: 277-282.
Okumus I., Mazlum M. (2002). Evaluation of commercial trout feeds: Feed consumption,
growth, feed conversion, carcass composition and bio-economic analysis. Tur. J. of
Fish.and Aqua Sci. 2:101-107.
Omoregie E.(2001). Utilization and nutrient digestibility of mango seeds and palm Kernel meal
by juvenile Labeosenegalenis (Antheriniformes: Cyprinidae). Aquaculture Research.
32:681-687.
Omoregie E., Igoche L., Ojobe T. O., Absalom K. V., Onusiriuka B. C. (2009). Effect of varying
levels of sweet potato (Ipomeabatatas) peels on growth, feed utilization and some
biochemical responses of the Cichlid(Oreochromis niloticus ). African Journal of Food
Agri., Nutrition and Development.9(2):1-7
Pachanawan A., Phumkhachorn P.,Rattanachaikunsopon P. (2008). Potential of
Psidiumguajavasupplemented fish diets in controlling Aeromonas hydrophilainfection in
tilapia (Oreochromis niloticus ). Journal of Bioscience and Bioengineering. 106:419-424.
27
Plaza I., García J. L., Villarroel M. (2018).Effect of spirulina (Arthrospira platensis)

supplementation on tilapia (Oreochromis niloticus ) growth and stress responsiveness
under hypoxia.Spanish Journal of Agricultural Research, 16(1):1-8..
Rana K. M., Salam M. A., Ahmmed M. R., Noor A. M.(2020).Dietary supplementation of
wheatgrass powder to assess somatic response of juvenile grass carp
(Ctenopharyngodonidella). Asian J. Med. Biol. Res. 2020, 6 (3), 482-490.
Rodge S. G., Thakare V. G., Joshi P. S. (2018).Study of dietary garlic induced effects on
hematology and biochemistry of Clarias batrachus (Linnaeus, 1758). Bioscience
Discovery, 9(3): 411-415.
Roghieh S., Seyed H., Maryam D., MasoudS. , Mina R. (2019).The effects
of Coriandrumsativum L. as feed additive on mucosal immune parameters, antioxidant
defence and, immune‐related genes expression in zebrafish (Daniorerio). 19:1-8.
Sahu S., Das B., Mishra B., Pradhan J., Sarangi N. (2007). Effect of Allium sativum on the
immunity and survival of Labeo rohita infected with Aeromonas hydrophila. J. Appl.
Ichthyol. 23:80-86.
Sareen M., Baghla P., Dhaka P., Mathur E., Sobti P., Khajuria S. (2014).Wheat Grass-A Wonder
Herb.Sys. Rev. Pharm. 5 (1): 4-5.
Shakya S. R. (2017). Effect of Herbs and Herbal Products Feed Supplements on Growth in
Fishes: A Review, Nepal Journal of Biotechnology. 5(1):58-63.
Shalaby S. M. M. (2004). Response of Nile tilapia, Oreochromis niloticus, fingerlings
to diets supplemented with different levels of fenugreek seeds (Hulba). J Agric Mansoura
Univ. 29:2231–2242.
Sharma A., Deo A. D., Riteshkumar S. T., Chanu T. I., Das A. (2010).Effect of Withania
somnifera(L Dunal) root as a feed additive on immunological parameters and disease
resistance to Aeromonas hydrophilain Labeo rohita(Hamilton) fingerlings.Fish and
Shellfish Immunology.29:508-512.
Sharma R., Nair A., Harak S., Patil T., Shelke S. (2016). Wheat Grass Juice: Nature‘s Powerful
Medicine. World J. Pharma.Pharmac.Sci.5(7): 384-391.
Sivagurunahan A., Xavier B., Gurusaraswathi S., Mariappan A. (2012).Immunostimulatory
potential of dietary amlaPhyllanthusemblicain growth and hematology of Tilapia
mossambicus challenged with Pseudomonas aeruginosa. International Research Journal
of Pharmacy. 3(7): 165-168
Skidmore-Roth L. (2003).Handbook of Herbs and Natural Supplements.2nd Edn. St. Louis:
Mosby. pp. 265.
Suriyavathana M, Roopavathi I and V Vijayan (2016).Phytochemical Characterization of
Triticum aestivum (Wheat Grass).J. Pharma. Phytochem.,5(1): 283-286.
28
(ISBN: 978-93-91768-45-4)
Thy S., Khieu B.,Vanvuth T., Thomas R P. (2008).Effect of water spinach and duckweed on fish
growth performance in poly-culture ponds.Livestock Research for Rural
Development 20(1): 1-6.
Turan F. (2006), Improvement of growth performance in tilapia (Oreochromis aureus Linnaeus)
by supplementation of red clover Trifolium pretense in diets. Israeli J Aquacult.58:34-38.
Uraiwan W., Wattana W., Karun T. (2019). Optimal Replacement of Fish Meal Protein by Stick
Water in Diet of Sex-Reversed Nile Tilapia (Oreochromis niloticus).Animals 9:521-533.
Won K. M., Kim P. K., Lee S. H., Park S. I. (2008).Effect of the residum extract of Siberian
ginseng Eleutherococcussenticosuson non-specific immunity in olive flounder
Paralichthysolivaceus. Fisheries Science.74:635- 641.
Yilmaz S., Ergün S., Türk N. (2012). Effects of herbal supplements on growth performance of
sea bass (Dicentrarchus labrax): Change in body composition and some blood
parameters. J BioSci Biotech.1(3):217-222.
Yin G., Jeney G., Racz T., Xu P., Jun X., Jeney Z. (2006).Effect of two Chinese herbs
(Astragalus radix and Scutellaria radix) on non-specific immune response of tilapia,
Oreochromis niloticus , Aquaculture.253:39-47.
YuanC.,PanX.,GongY.,XiaA.,WuG.,TangJ. (2008).Effectsof Astragalus polysaccharides (APS)
on the expression of immuneresponse genes in head kidney, gill and spleen of the common
carp, Cyprinus carpio L. International Immunopharma.;8:51-58.
Zakes Z., Kowalska A., Demska-Zakes K., Jeney G., Jeney Z. (2008). Effect of two medicinal
herbs (Astragalus radix and Lonicera japonica) on the growth performance and body
composition of juvenile pike perch (Sander lucioperca). Aquacult.39:1149-1160.
Zakes Z., Kowalska A., Demska-Zakes K., Jeney G., Jeney Z. (2008). Effect of two medicinal
herbs (Astragalus radix and Lonicera japonica) on the growth performance and body
composition of juvenile pike perch (Sander lucioperca). Aquacult.39:1149-1160.
29
PRODUCTION OF VACCINE AGAINST GASTROINTESTINAL NEMATODE

HELIGMOSOMOIDES POLYGYRUS
Dipti Bhimrao Kadu
Department of Zoology,
Arts and Science College, Pulgaon, Maharashtra
Corresponding author E-mail: [email protected]
Introduction:
Heligmosomoides polygyrus is a natural intestinal parasite of mice. Chronic helminth
infections remain a huge global health problem. It causes extensive deaths in both humans and
livestock. Some mouse strains evade immunity to the parasite naturally while in some it is
induced through immunization. The mechanisms of protective immunity are not yet fully
defined. Humans are hosts to nearly 300 species of parasitic worms and over 70 species of
protozoa. There are number of misconception about parasitic infections that it occurs only in
tropical areas but many people in temperate and subtropical areas also become infected. This
statement was proposed by parasitologist F.E.G. Cox that some parasitic infection was derived
from our primate ancestors and some acquired from our domestic animals or we can get infection
from contact with these parasites during our relatively short history on Earth.
Materials and Methods:

Experimental animal:
The Swiss albino mouse, Mus musculus albinus of either sex were selected as an
experimental animal for the present investigation. The mice were obtained from the Institute of
Nutrition (NIN) Hyderabad, India and were kept in the animal house under local conditions of
light, temperature ventilation and food. Food and water were provided ad libitum. Male and
female healthy mice of 6-8 weeks old and 15-20 gms in weight were used according to the need
of the experimental design. Animal experimentations were conducted according to INSA ethical
guidelines for the use of animals for scientific research purpose, after permission from the ethical
committee.
The Parasite: Heligmosomoides polygyrus
Experimental studies on Heligmosomoides polygyrus are few. H. polygyrus is a
trichostrongyloid nematode of family Heligmosomidae, first reported from Mus musculus by
30
(ISBN: 978-93-91768-45-4)
Spurlock (1943) and from Peromyscus maniculatus by Ehrenford (1954). H. polygyrus is a long-
lived intestinal nematode of mice. Rats are known to be naturally resistant to it (Cross, 1960).
Development of its free living stage has been reported by Fahmy (1956) and its complete life
cycle in mice by Spurlock (1943); Baker (1954); Ehrenford (1954); Dobson (1960); Callizo
(1962) and Bryant (1973). Adults of H. polygyrus are spirally coiled (females 18-21 mm and
males 8-10 mm long) and are found in first 2 cm. attached to duodenal mucosa. The eggs of H.
polygyrus are fully emryonated when evacuated with feces and hatch at 20-23° C. Life cycle is
direct and non- migratory, infection to host is through ingestion of infective filariform 3rd stage
larvae which penetrate intestinal mucosa maturing in 8-9 days after infection and females lay
eggs up to 9 months producing 1280 eggs per day per female (Scott, et al., 1959). Infection can
easily be maintained in the laboratory and the larvae culture well within limits and thus, forms an
excellent parasitological tool for experimental investigations.
Experimental protocol:
The mice were divided into following six groups –
1) Non Infected Non Vaccinated Control – 1
2) Infected Non –Vaccinated Control –2
3) Infected Vaccinated with Larval Somatic Antigens
4) Infected Vaccinated with Adult Somatic Antigens
5) Infected Vaccinated with Larval ES Antigens
6) Infected Vaccinated with Adult ES Antigens
Preparation of inoculums for infection:
A larval suspension of about 100 ml was prepared in a glass stoppered measuring
cylinder of 100 ml capacity. The numbers of actively motile larvae were counted by the dilution
method of Scott (1928). After vigorous shaking, 1 ml of the suspension was pipette out,
transferred onto several glass slides with squares already made on their reverse with a glass
marking pencil and the larvae in all squares were carefully counted under a suitable dissecting
microscope. Three such counts were repeated and the average count in 1 ml was multiplied by
the total volume to get the total number of the larvae. An inoculum containing the desired
number of actively motile larvae was adjusted in 0.2-0.3 ml to be given to each mouse. Each
mouse was orally inoculated directly into stomach with the desired number of larvae (300) by 1
ml syringe having a blunt 18 gauge-feeding needle. After inoculation, mice were kept in cages in
groups of five and labeled according to the design of experiments and were fed routinely with the
same standard diet.
31
Worm recovery:
1. L-3 larval and adult worm recoveries from infected mice (with 300 larvae of H.
polygyrus) after 1, 5 and 13th day of administration of larval somatic antigens:

Each mouse was infected with 300 larvae and after 1, 5 and 13 days of infection worm
recovery was seen. In infected non-vaccinated mice 29.01% recovery was observed after 5 days
and out of them 26.26% adults were recovered after 13 days.
While in mice which were vaccinated by larval somatic antigens, 9.41%, 6.20% and
5.22% recovery was recorded after 1, 5 and 13 days of vaccination [Table-4.1, Fig. 4.1].
polygyrus) after 1, 5 and 13th day of administration of adult somatic antigens:

In infected non-vaccinated mice 29.01% recovery was observed after 5 days and out of
them 26.26% adults were recovered after 13 days.
While in mice which were vaccinated by adult somatic antigens, 9.66%, 7.40% and
6.07% recovery was recorded after 1, 5 and 13 days of vaccination [Table-4.2, Fig. 4.2].
polygyrus) after 1, 5 and 13th day of administration of larval ES antigens:

them 26.66% adults were recovered after 13 days. While in mice which were vaccinated by
larval ES antigens, 6.03%, 5.26% and 3.95% recovery was recorded after 1, 5, and 13 days of
vaccination [Table-4.3, Fig. 4.3].
polygyrus) after 1, 5 and 13th day of administration of adult ES antigens:

them 26.66 % adults were recovered after 13 days. While in mice which were vaccinated by
adult ES antigens 8.21%, 6.85% and 5.36% recovery was recorded after 1, 5, and 13 days of
vaccination.
Observations and Results:

Heligmosomoides polygyrus is a natural intestinal helminth of mice. It inhibits in the
duodenal region of small intestine. In the present investigation, albino mice were colonized with
300 larvae each time by placing larvae directly in the stomach through gastric lavage. The larvae
migrated to duodenum, to house in the submucosa where they matured and then emerged as adult
worms migrating in the intestinal lumen by 13 days of time.
32
(ISBN: 978-93-91768-45-4)
The present study was carried out to investigate the immunological responses and
possibility of vaccine development.
Summary and Conclusion:

The % protection after administration of larval ES antigen was the highest followed by
vaccination with somatic antigens, then adult ES antigen and least after vaccinated with adult
somatic antigens. Larval ES > Larval somatic antigens > adult ES antigens > adult somatic
antigens.
140
120
WORM RECOVERIES
100
80 I-NV-C-2
69.78 %
80.12 %
60 I-V-LSA
78.69 %
40
20
0
1 5 13
Days after infection
Figure 1: L-3 larval and adult worm recoveries from infected mice (with 300 larvae of H.
polygyrus) after 1, 5 and 13 days of administration of larval somatic antigens
140
120
WORM RECOVERIES
100
80
I-NV-C-2
60 I-V-ASA
73.32 %
74.49 %
69 %
40
20
0
1 5 13
polygyrus) after 1, 5 and 13 day of administration of adult somatic antigens
33
I-NV-C-2 Infected Non Vaccinated Control-2

I-V-ASA Infected and Vaccinated with adult Somatic Antigens
I-V-LSA Infected and Vaccinated with L-3 larval Somatic Antigens
Values on I-V-LSA bar denote % protection
Values on I-V-ASA bar denote % protection
140
120
100
WORM RECOVERIES
80
I-NV-C-2
60 I-V-LESA
80.63 %
81.90 %
84.96 %
40
20
0
1 5 13
Figuure 3: L-3 larval and adult worm recoveries from infected mice (with 300 larvae of H.
polygyrus) after 1, 5 and 13 day of administration of larval ES antigens
140
120
100
WORM RECOVERIES
80
I-NV-C-2
60 I-V-AESA
73.63 %
76.46 %
40
79.59 %
20
0
1 5 13
polygyrus) after 1, 5 and 13 day of administration of adult ES antigens
I-NV-C-2 Infected Non Vaccinated Control-2
I-V-LES Infected and Vaccinated with L-3 larval Somatic Antigens
34
(ISBN: 978-93-91768-45-4)
I-V-AES Infected and Vaccinated with adult ES Antigens

Values on the I-V-LESA bar denote percent recovery
Values on the I-V-AESA bar denote percent recovery
After 1, 5 and 13 days of administration of larval ES antigens, the protection achieved

was 80.63%, 81.90% and 84.96% respectively which was quiet better than the rest of the
antigens tried during the present investigation. Even after the 1st day of administration of larval
ES antigens, the larval recovery was 80.63%, while in rest of the vaccinated conditions the
protection was below 75%. Vaccination with larval antigens, both somatic and ES antigens, led
to less recovery of mature worms, than vaccination after adult somatic and ES antigens and
similarly ES antigens were more potent than somatic antigens. These differences may be due to
developmental modifications in larvae to become adult for continuation of race.
References:
Adams, J. H.; East, I. J.; Monroy, F.; Washington, E. A. and Dobson, C. (1987). Stage-specific
antigens of Nematospiroides dubius Baylis, 1926 (Nematoda: Heligmosomidae). J.
Parasitol. 73, 1164-8.
Anthony, Robert M.; Laura, I.; Rutitzk.; Joseph F.; Urban, Jr.; Miguel J.; Stadecker and William,
C. Gause (2007). Protective immune mechanisms in helminth infection. Nat. Rev.
Immunol. 7 (12), 975-987.
Baggiolini, M.; Dewald, B.; Moser, B. (1994). Interleukin-8 and related chemotactic
cytokines: cxc and cc chemokines. Adv Immunol. 55, 97-179.
Baker, N. F. (1954). Trichostrongylidosis: The mouse as an experimental animal. Pro. Am.Vet.
Med. As. 91, 185-191.
Bryant, V. (1973). The life cycle of Nematospiroides dubius, Baylis,1926 (Nematoda:
Heligmosomidae). J. Helminth. 47, 263-268.
Callizo, P. J. (1962). The free living development of Nematospiroides dubius, Baylis, 1926
(Nematoda: Heligmosomidae).
Cooper P. J.; Chico, M. E.; Losonsky, G. et al., (2000). Albendazole treatment of children with
ascariasis enhances the vibriocidal antibody response to the live attenuated oral cholera
vaccine CVD 103-HgR. J Infect Dis. 182, 1199–1206.
Crawford, C.; Behnke, J. M. and Pritchard, D. I. (1989). Suppression of heterologous immunity
by Nematospiroides dubius antigens in vitro. Int. J. Parasitol. 19, 29-34.
35
Cross, J. H. Jr. (1960). The natural resistance of the white rat to Nematospiroides dubius and the
effect of cortisone on this resistance. J. Parasit. 46, 175-185.
Cypess, R. H. and Zidian, J. L. (1975). Heligmosomoides polygyrus (Nematospiroides dubius):
the development of self-cure and/or protection in several strains of mice. J. Parasit. 61,
819-824.
Donald, A. D. and Waller, P. J. (1982). Problems and prospects in the control of helminthiasis in
sheep. Page 157 in Biology and Control of Endoparasites. L. E. A. S ymons, A. D. Donald,
and J. K. Dineen, ed. Academic Press, New York, NY.
Dobson, C. (1960). ―An investigation of the host parasite relations and host specificity of
Nematospiroides dubius Baylis, 1926, Heligmosomadae, a mouse nematode in its normal
and abnormal hosts‖. Ph.D. thesis, University of Scheffield.
Ehrenford, F. A. (1954). The life cycle of Nematospiroides dubius, Baylis, 1926 (Nematoda:
Heligmosomidae). J. Parasit. 79, 480-481.
Fakae, B. B.; Harrison, L. J.; Sewell, M. M. (2000). The intensity and duration of primary
Heligmosomoides polygyrus infection in TO mice modify acquired immunity to secondary
challenge. Immunology. 43, 56-60
Fahmy, M. A. M. (1956). An investigation on the life cycle of Nematospiroides dubius
(Nematoda: Heligmosomidae) with reference to the free living stages.Z. Parasiten. 17, 394-
399.
Garside, P.; Kennedy, M. W.; Wakelin, D. and Lawrence, K. E. (2000). Immunopathology of
intestinal helminth infection. Parasite Immunology. 22, 605-612.
Kerboeuf, D. and Jolivet, G. (1980). Repeated anthelmintic treatments and receptivity of mice to
experimental infections with Heligmosomides polygyrus (Nematospiroides dubius). Ann.
Rech. Vet. 11, 185-93.
Scott, J. A.; Cross, J. H. Jr. and Dawson, C. (1959). Egg production of Nematospiroides dubius
in mice and rats. Texas Reports on Biol. Med. 17, 610-617.
Spurlock, G. M. (1943). Observation on the host parasite relations between laboratory mice and
Nematospiroides dubius. Baylis. J. Parasit. 29, 303-311.
Robinson, M.; Gustad, T. R. and Erickson, M. R. (1997). Immunological interactions between
Trichinella spiralis and Heligmosomoides polygyrus: cross reactivity between muscle
larvae and antibodies raised to unrelated antigens. Int. J. Parasitol. 27, 865-72.
Yazdanbakhsh, M.; van den Biggelaar, A.; Maizels, R. M. (2001). Th2 responses without atopy:
immunoregulation in chronic helminth infections and reduced allergic disease. Trends
Immunol. 22, 372-377.
36
(ISBN: 978-93-91768-45-4)
TECHNOLOGICAL ADVANCES AND ANIMAL HEALTH

Mohsina Mushtaq
Department of Veterinary Medicine,
SKUAST-K, Shuhama, Kashmir, India
Abstract:
In this rapidly growing world of technology new techniques are being used for increasing
animal production, efficiency and profitability as well as to provide more comfort to farmers.
Due to increasing market pressure dairy industry is moving towards more intensive, profit driven
enterprise which in turn demands use of modern technologies that are cost effective and provide
maximum benefits to the industry. As animal diseases cost the livestock industry billions of
dollars each year new vaccines, disease monitoring systems and diagnostic kits can help to
decrease both morbidity and mortality levels in livestock.Application of biotechnology to the
livestock industry has generated a number of products for improving production, animal health,
and food processing.The aim of this review is to focus on the modern technologies that are being
currently used for improving animal health and increasing national economy through livestock
sector.
Keywords: Advances, Animal health, Technologies
Introduction:
Animal health is important to the farmers as well as to the livestock and dairy industry of
the country for gaining maximum benefits as livestock sector is the main component of India‘s
agrarian economy providing food and nutritional security and livelihood. Over the past few years
advanced digital technologies have improved every sector of the economy, including animal
production, health and welfare. These include wireless and mobile technologies for animal health
monitoring, disease surveillance, reporting and information sharing; advanced data processing
technologies such as big data and data analytics used to uncover hidden patterns, predictions,
correlation and other information; and promising technologies such as blockchain applications
used for effective and efficient management of various input supply chains.In order to meet the
food production needs due to increasing world population,livestock sector has to satisfy the
increasing world consumption of animal-source food through sustainable animal production in
37
ways that promote food security, poverty reduction, public health and food safety (FAO, 2009
and Tedeschi et al., 2015).
In addition to the increased world population and its demand for more reliable quality
livestock products the livestock production problems are also increasing (Thornton, 2010). As
the number of animals increase error burden and work load also increases. Hence for better
production successful livestock farmers rapidly adapt their infrastructures with changes in
technology. As such, automation systems provide options in front of the user in intense
competition for convenience. Manual observation is gradually beingreplaced by many milking
systems by automated recording (milk yield, milk conductivityand other measurements) leading
to better quantity and quality of data.
Electronic recording, milking, heat detection auto-weighing,auto-drafting, genetic
improvement, feeding, barn optimization, and health monitoring, livestockhousing and
equipment designs are the technologies that provide dairyman many opportunitiesto make easier
and more convenient decisions about dairy future plans.
Modern technologies in animal science use:

Biotechnology:
It involves techniquessuch as genetic engineering, cell culture, and monoclinal antibody
methods that use living organisms or their partsto make or modify products, to improve plants or
animals,or to develop microorganisms for specific uses. It is specifically used to produce
productsthat promote growth, increase feed efficiency,carcass leanness in growing animals, and
significantlyincrease milk production in lactating animals.Transgenic animals producedgrow
faster, have greater disease resistance, and produce high-value pharmaceutical products. Also,
newvaccines and diagnostic kits are being developed to improvelivestock health. Biotechnology
is also being usedto process meat and dairy products and to detect foodcontaminants that might
be present in those products.
Anabolic steroids and antimicrobial compounds:
These are currently being used in the livestock sector topromote growth and increase feed
efficiency. However, new products are also being produced like protein hormones
(somatotropins) and catecholamine compounds (beta-adrenergicagents) that increase growth
rates inyoung animals, improves FCE (food conversion efficiency) and significantly reduce
carcassfat so that meat products are leaner. Somatotropins also increase milk production in
lactating dairy cows.
38
(ISBN: 978-93-91768-45-4)
Reproduction technologies:
Like estrous cycle regulation, embryo cloning, superovulation, embryo and sperm sexing,
ETT (embryo transfer technology) and production of transgenic animalsare being used to
improve the quality of livestock herds more rapidly than could be achieved with traditional
breeding.
Animal health technologies:
These are being used to create new vaccines which include deleting or inactivating the
genes in a pathogen that cause disease,and inserting into a vector gene that cause an
immuneresponse to a pathogen. Synthetic peptides are also beingproduced that stimulate the
immune response like genetically engineered protein lysostaphin kills Staphylococcus aureus
and achieves cure rates as high as 80 percent for mastitis in animals.Some synthetic peptides
have been used to inhibit critical functionsof lentiviruses in sheep. Techniques for advanced
laboratory diagnosis include enzyme immunoassays (ELISA), monoclonal antibodies,
recombinant antigens, immunoblotting, nucleic acid hybridization, PCRand real time PCR
techniques and nucleic acid analysis.Advances in vaccine technology aims to improve the
immunogenicity and safety of vaccines, their shelf life and their cost of production. These
techniques include recombinant DNA technology, modified live virus vaccines, synthetic
peptides as vaccines, sub-unit vaccines, vectored vaccines, chimeric live vaccines and DNA
vaccines. Immunomodulators like lymphokines (interleukins and interferons) and cytokines are
hormone-like molecules that are being used for coordinating immune defences to infectious
agents, cancer, and autoimmune diseases.
Digital technologies:
It includes new methods of data collection and management using advanced information
and communication technologies (ICTs) andinnovations. Recent developments in ICTs and
innovations lead to newopportunities for improving veterinary practice (Bellet, 2019 and EBVS,
2019), timeliness and accuracy of datacollection and reporting for diseasesurveillance and animal
health monitoring (Holmstrom and Beckham, 2017). The use ofnew ICTs also facilitate mapping
and monitoring the spread of infectious diseases and their coordination across sectors, as well as
tracking supplies of drugs and vaccines (WHO, 2018). These developments lead to better, more
efficient, and timely decisions that affect the performance and quality of veterinary services
meeting standards of animal health and welfare practices (Liu et al, 2019). Mobile phones are
being used for data collection onanimal diseases surveillance in animal health (Robertson et al.,
2010 and Madder et al., 2012). In veterinary epidemiology, big data analysisresults in better
39
understanding of animal diseases and health related risks and improvements in the fields of
bioinformatics which facilitate the understanding of host-pathogen interactions towards the
development of new diagnostics, therapeutics and vaccines (Deblais et al., 2019). In case of
livestock and veterinary sectors, blockchain-based systems are potentially applied for efficient
management of various input supply chains such as animal feed, veterinary drugs, diagnostic kits
and vaccines (Makkar and Costa, 2020).
Sensor technology:
It is an effective method for health monitoring which uses a sensor for measuring
physiological or behavioural parameter of an individual cow and enables automated, on-farm
detection of changes in this condition that is related to a health event like a disease and requires
action on the part of the farmer such as treatment. Sensors are of two types: Attached and Non-
attached and currently the sensor-based data acquisition systems are classified into two
categories: Non-invasive (Immobile sensors located in the barn and Mobile sensor boxes
attached to the cow called external sensors) and Invasive.
Examples of non-invasive type of sensors include temperature measurements of the udder
or of the face in an automatic milking station and measurement of breath composition.
Surveillance cameras are another kind of immobile sensors that can continuously provide
information for the cows in the herd (Poursaberi et al., 2010). The most reliable way to monitor
cows throughout a day is to attach sensors at individual cow by a neck collar or an ankle ribbon.
Typical sensors of this type are accelerometers, pedometers, vibration sensors,
thermometers for temperature measurements, humidity sensors etc. Pedometers are cheap and
simple sensors that give insight in the activity status of a cow like oestrus behaviour with good
prediction capabilities (Løvendahl and Chagunda, 2010).Recently, low-cost and infrastructure-
less GPS positioning sensorshave been used to identify different motionstates of cows (Godsk
and Kjærgaard, 2011). The GPS sensors attached to the animal‘s collars identify the following
activities: eating, seeking, walking, lying and standing.
Invasive sensors include mobile sensor boxes swallowed or implanted to cow called
internal sensors.Typical sensors of this kind are thermometers for measuring the core body
temperature or vaginal pressure during birth, sensors for measuring electrical conductivity and
pH value of rumen fluid.
Advanced Animal Breeding and Genetics:
Innovations in animal breeding and genetics result in improving food quality. Genome
editing techniques in animals has led to greater meat and milk production. Various approaches to
retrieve superior qualities in animal breeds include artificial insemination, somatic cell nuclear
40
(ISBN: 978-93-91768-45-4)
transfer, in vitro production of embryos, gene transfer, nuclear transfer and aquaculture.New
animal breeds have been developed in animal husbandrywith the help of breeding and gene
technology. Till 1980s livestock products demands have been met by breed substitution, cross-
breeding, and within-breed selection. But these demands in future are to be met using new
techniques such as artificial insemination and more specific selection techniques.
Conclusion:
Recent advances in animal health are a result of the application of new technologies in
the field of animal husbandry and veterinary sciences. New technological interventions and
application of digitalization in animal breeding and genetics, biotechnology, animal disease
monitoring and survillence, vaccine production and other prophylactic and therapeutic
approaches for improvement of animal health which in turn has increased the productivity and
economic status of the country to a great extent. However, more is to be done to improve the
health, productivity and efficiency of the livestock for gaining maximum benefits.
References:
Bellet, C. 2019. The Future of Animal Health: How Digital Technologies Reconfigure Animal
Healthcare in Farming. Discover Society, 7 August.
Deblais, L., Kathayat, D., Helmy, Y.A., Closs, Jr. G. and Rajashekara, G. 2019. Translating ‘big
data’: better understanding of host-pathogen interactions to control bacterial foodborne
pathogens in poultry. Anim. Health Res. Rev., 21 (1), 15–35.
European Board of Veterinary Specialisation. 2019. ECCVT workshop Embracing digital
technology in veterinary practice Brussels, 13th and 14th May 2019. EBVS, Thessaloniki,
Greece.
Food and Agriculture Organization of the United Nations. 2009. The State of Food and
Agriculture: Livestock in the balance. FAO, Rome, Italy, 180 pp.
Godsk, T. and Kjærgaard, M. B. 2011. High classification rates for continuous cow activity
recognition using low-cost GPS positioning sensors and standard machine learning
techniques, pp. 174–188.
Holmstrom L.K. and Beckham T.R. 2017. Technologies for capturing and analysing animal
health data in near real time. In Biological threat reduction (T. Beckham, ed.). Rev.Sci.
Tech. Off. Int. Epiz. 36 (2), 525–538.
41
Liu, J., Toma, L., Barnes, A.P. and Stott, A. 2019. Farmers uptake of animal health and welfare
technological innovations. Implications for animal health policies. Front. Vet. Sci. 6: 410.
Løvendahl, P. and Chagunda, M. G. G. 2010. On the use of physical activity monitoring for
estrous detection in dairy cows., J. Dairy Sci. 93 (1): 249–59.
Madder, M., Walker, J.G., Van, Rooyen J., Knobel, D., Vandamme, E., Berkvens, D.,
Vanwambeke, S.O. and De Clercq, E.M. 2012. e-Surveillance in animal health: use and
evaluation of mobile tools. Parasitology. 139 (14): 1831–1842.
Makkar, H.P.S. and Costa, C. 2020. Potential blockchain applications in animal production and
health sector. CAB Rev. 15 (035): 1–8.
Poursaberi, A., Bahr, C., Pluk, A., Van Nuffel, A. and Berckmans, D. 2010. “Real-time
automatic lameness detection based on back posture extraction in dairy cattle: Shape
analysis of cow with image processing techniques, Comput. Electron. Agric. 74 (1): 110–
119.
Tedeschi, L.O., Muir, J.P., Riley, D.G. and Fox, D.G. 2015. The role of ruminant animals in
sustainable livestock intensification programs. Int. J. Sustainable Dev. World Ecol. 22 (5),
452–465.
Thornton, P.K. 2010. Livestock production: Recent trends, future prospects. Philosophical
Transactions of the Royal Society, B: Biological Sciences. 365 (1554): 2853-2867.
World Health Organization. 2018. Digital technologies: shaping the future of primary health
care. WHO, Geneva, Switzerland.
42
(ISBN: 978-93-91768-45-4)
CLONING AND GENETIC ENGINEERING

Khan Rumana Amanullah
J. A. T. Arts, Science and Commerce College (For Women), Malegaon
Abstract:
The technique of creating a genetically identical replica of a cell or an organism is known
as cloning. Naturally cloning occurs when a cell copies itself asexually i.e. without undergoing
any genetic alterations or recombination. Bacteria and other prokaryotic organisms (those
without a cell nucleus) use binary fission or budding to generate genetically identical replicas of
them. With the exception of gametes (eggs and sperm), which undergo meiosis and genetic
recombination, all cells that undergo mitosis, such as skin cells and cells lining the
gastrointestinal system, are clones in eukaryotic beings (organisms with a cell nucleus).
Cloning is a wide term in biomedical research that refers to the duplication of any type of
biological material for scientific study, such as a bit of DNA or a single cell. Segments of DNA,
for example, are reproduced exponentially using a technique known as polymerase chain
reaction, or PCR, which is extensively employed in fundamental scientific research. The
development of cloned embryos, particularly human embryos, that are genetically identical to the
species from which they are derived, and their subsequent usage for scientific, medicinal, or
reproductive purposes is the topic of heated discussion.
Reproductive cloning:
The purposeful generation of genetically identical people is known as reproductive
cloning. Each new individual is exactly same as the original. Natural clones are monozygotic
(identical) twins. Hence the genes and the DNA present in the nucleus of cells from two clones
are identical. A method for creating a clone or an exact replica of a whole multicellular organism
is reproductive cloning. The majority of multicellular creatures reproduce sexually, which entails
the genetic hybridization of two individuals (parents), making it difficult to produce an exact
duplicate or clone of either parent. Asexual reproduction of animals may now be intentionally
induced in the laboratory thanks to recent breakthroughs in biotechnology.
Parthenogenesis, or "virgin birth," is a type of asexual reproduction in which an embryo
grows and develops without the egg being fertilised. In animals where the female deposits an
egg, parthenogenesis is an example. If the egg is fertilised, it becomes a diploid egg and the
individual becomes a female; if the egg is not fertilised, it becomes a haploid egg and the
43
individual becomes a male. A parthenogenic, or virgin, egg is one that has not been fertilised.
Parthenogenic eggs, which can grow into adults, are laid by some insects and reptiles.
For sexual reproduction, it is necessary to have two cells. When the haploid egg and
sperm cells unite it forms a diploid zygote. The genetic information that is required to create a
new human is stored in the nucleus of zygote. Early embryonic development, on the other hand,
needs the cytoplasmic material present in the egg cell. Reproductive cloning is based on this
concept. When the haploid nucleus of an egg cell is replaced with a diploid nucleus from any
member of the same species (donor), the result is a zygote with the same genetic makeup as the
donor. The procedure of putting a diploid nucleus into an enucleated egg is known as somatic
cell nuclear transfer. It's suitable for both reproductive and therapeutic cloning.
Dolly, a sheep born in 1996, was the first cloned mammal. At the time, reproductive
cloning had an extremely low success rate. Dolly survived for seven years before succumbing to
respiratory problems. It's possible that because the cell DNA belongs to an older person, the age
of the DNA will impact the lifespan of a cloned person. Several animals (including horses, bulls,
and goats) have been successfully cloned since Dolly, albeit these creatures frequently have
facial, limb, and cardiac defects. Attempts to produce cloned human embryos as sources of
embryonic stem cells have been made. The procedure, which is sometimes referred to as
therapeutic cloning, creates stem cells that are used to treat illnesses or abnormalities (unlike
reproductive cloning, which aims to reproduce an organism). Despite this, therapeutic cloning
initiatives have been faced with opposition due to bioethical concerns.
44
(ISBN: 978-93-91768-45-4)
Molecular cloning:
Molecular cloning is a collection of molecular biology experimental methods for
assembling recombinant DNA molecules and directing their replication in host organisms.
Cloning refers to a technique that involves the reproduction of a single molecule to generate a
population of cells with identical DNA molecules. In most cases, DNA sequences from two
separate animals are used in molecular cloning: the source of the cloned DNA and the species
that will act as the live host for recombinant DNA replication. Many contemporary fields of
modern biology and medicine rely on molecular cloning procedures.
In a traditional molecular cloning procedure, the DNA to be cloned is acquired from a
target organism and then processed with enzymes in the test tube to produce smaller DNA
fragments. After that, these pieces are mixed with vector DNA to make recombinant DNA
molecules. After that, the recombinant DNA is implanted into a host organism (typically an easy-
to-grow, benign, laboratory strain of E. coli bacteria). This will result in a colony of creatures
that reproduce recombinant DNA molecules alongside host DNA. These are transgenic or
genetically engineered bacteria because they contain foreign DNA pieces (GMO). The fact that a
single bacterial cell may be encouraged to take in and reproduce a single recombinant DNA
molecule is used in this technique. This single cell may then be multiplied indefinitely to produce
a vast number of bacteria, each containing a copy of the original recombinant molecule. As a
result, both the recombinant DNA molecule and the ensuing bacterial population are usually
referred to as "clones." Recombinant DNA, strictly speaking, relates to DNA molecules, whereas
molecular cloning refers to the experimental procedures used to put them together. Different
DNA sequences may be put into a plasmid, and these alien sequences would be taken into
bacteria and digested as part of the plasmid, according to the theory. In other words, these
plasmids might be employed as cloning vectors for carrying genes.
Although virtually any DNA sequence may be cloned and amplified, various variables
may restrict the procedure' effectiveness. Inverted repeats, replication origins, centromeres, and
telomeres are examples of difficult-to-clone DNA sequences. When inserting large-sized DNA
sequences, there is also a decreased possibility of success. Insertions bigger than 10 kbp have a
low success rate, yet bacteriophages like bacteriophage can be engineered to insert sequences up
to 40 kbp effectively.
Therapeutic cloning:
Therapeutic cloning refers to the cloning with the purpose of treating a sickness. The
nucleus of a cell, usually a skin cell, is put into a fertilised egg whose nucleus has been removed
45
in therapeutic cloning. A blastocyst is formed when a nucleated egg divides repeatedly. The
blastocyst's stem cells are then extracted and used to generate cells that are genetically identical
to the patient. Therapeutic cloning cells can then be put into a patient to address a condition that
they are suffering from. In contrast to therapeutic cloning, the purpose of reproductive cloning is
to generate a new individual, a proposition that has sparked widespread debate and widespread
opposition.
Therapeutic cloning might result in stem cells that are genetically identical to the patient.
The procedure entails transferring the nucleus from a patient's cell to an egg cell that has had its
nucleus removed.
Benefits and risks associated with the use of stem cells in medicine. Stem cells show a lot
of promise in terms of treating patients with diseases that are now incurable, generating organs
for transplantation, and research. However, its usage raises clinical, ethical, and societal
concerns. These challenges will be different for adult, embryonic, and therapeutically-cloned
stem cell development and transplantation. They will also be determined by whether the stem
cells will be employed for therapeutic or research purposes.
It's critical to get a balanced perspective. There are no right or wrong solutions or even
answers at all, in certain cases. When talking about stem cells, there are a few things to think
about:
46
(ISBN: 978-93-91768-45-4)
Clinical problems:
 There is no assurance that these therapies, such as the use of stem cells to replace nerve
cells lost in Parkinson's disease patients, would be successful.
 The challenge in identifying acceptable stem cell donors at the moment.
 The difficulty of acquiring and preserving embryonic stem cells from a patient. These
would have to be obtained before to delivery; some clinics will retain blood from a
newborn's umbilical cord.
 In stem cells cultivated for several generations, mutations have been discovered, and some
altered stem cells have been seen to behave like cancer cells.
 Viruses might infect cultured stem cells, which could then be transmitted to a patient.
Ethical concerns:
 Is it ethical to make embryos for therapeutic cloning and then kill them?
 Are unused embryos created by in vitro fertilization (IVF) a source of embryonic stem
cells?
 Embryos might be considered as a commodity rather than an embryo that has the potential
to develop into a person.
 At what point in its development should an embryo be considered a person and treated as
such?
Social Concerns:
 It is critical to educate the public on what stem cells can and cannot do.
 Whether the advantages of stem cell therapy outweigh the drawbacks.
 Because a lot of the research is done by for-profit clinics, the results aren't subjected to
peer review. Because stem cell therapies are still in their early phases of research, patients
may be abused by paying for expensive procedures and being offered false hope of a cure.
Conclusion:
The same scientific procedures that allow us to clone animals may be used to duplicate
particular cells throughout the body. The techniques established in this sector might eventually
be used to create new tissues or organs as needed. It would not endanger the animal's life, and
the knowledge gathered from these procedures might lead to new advances in human medical
research. We might analyze the cloned cells to see how effective each procedure is in
determining how these fields can advance. Cloning animals might provide better comfort to pet
owners.
47
Losing a pet is especially painful since these creatures are generally considered family
members. Our everyday routines are made more structured by the presence of cats, dogs, and
other animals. These partners keep us busy and might even encourage us in our attempts to
conquer life's obstacles.
Pets provide us with a sense of purpose. Cloning animals would allow individuals to
safeguard their memories of a valued friend by having a scientist make an identical animal. This
endeavour would still result in a one-of-a-kind animal for the species, but it would also provide
some continuity and support, potentially reducing emotional reactions. Animal cloning does not
result in precise duplication.
When we clone animals, we do not achieve precise replication:
The genetic material is implanted into an embryo, which is made up of distinct cells. The
clone can then create children later in life as a result of these procedures. Because Dolly the
Sheep was coupled with a Welsh Mountain ram, scientists were able to produce six lambs in all.
Bonnie was the first, and she was born in 1998.
Then Dolly gave birth to twins named Rosie and Sally by her carers. Cotton, Darcy, and
Lucy were the names of her triplets, who were born in the fall of 2001. Some experts believed
that the children would be infertile, but they were not, demonstrating the full potential of this
scientific method.
Animal cloning helps us to save endangered species:
Only because 13 horses were caught from a wild herd and maintained in a zoo in the
1940s were researchers able to conserve the Przewalski's horse. Two of the horses were hybrids
even back then. Standard breeding procedures aided in the survival of the species, which
currently numbers in the thousands.
With the Northern White Rhino, we won't have the same luck. In March 2018, the
species' only known male died. Because there are only two females of the same subspecies left
alive, cloning is the only method to rescue them. This work allows us to save endangered
creatures and maybe even bring extinct species back into the world.
Cloning animals allows us to develop the most desirable characteristics:
Through cloning scientists can do what other scientists have done for more than 1000
years through selective breeding. The results are comparable to those obtained through natural
reproductive processes involving human intervention. This approach offers the opportunity to
develop exact, desirable features in animals.
Animal cloning might be used to create dairy cows that generate more milk. Researchers
may investigate the possibility of cloning certain chickens to boost commercial egg output. This
48
(ISBN: 978-93-91768-45-4)
method might be used to breed livestock animals to generate more meat per corpse. When we
consider the whole potential of this technology, the possibilities are practically endless
Human diseases could be reduces with the help of animal cloning:
Influenza is one of the most troublesome infections that humans face each season.
Millions of people have died as a result of flu epidemics in the past, especially when a new strain
of the virus spreads across the population. Even though yearly immunizations are available to
prevent it, about one in every five people will contract it each year.
The influenza virus is spread via birds, pigs, and other animals. Our animal cloning techniques
may be able to halt its development by increasing resistance to its activities as it develops. It's an
opportunity to avert disease's negative consequences before it even begins.
Quality of food supply would not be altered by cloning:
People can consume animal products from cloned animals, according to 2008 rulings
based on science at the time. Any animal species can enter the commercial food chain, according
to the FDA. That means scientists may seek for methods to improve the nutrient profile of the
proteins we eat, giving everyone the opportunity to eat better without having to change their
eating habits. Consider this benefit to be the beef equivalent of working with enriched flour.
References:
Blackman, K. (2001) The coming of hereditary designing. Patterns in Biochemical Science, 26,
268–270. (An record of the beginning of quality cloning.)
Brown, T.A. (2006) Genomes, third edn. Festoon Science, Oxford. (An prologue to current
hereditary qualities and atomic biology.)
Colman A. Molecular Cell Biology. fourth edition. Physical cell atomic exchange in warm
blooded animals: Progress and applications . Cloning 1999, 1(4): 185-200. (PubMed)
Dale, J.W. and Park, S.T. (2004) Molecular Genetics of Bacteria, fourth edn. Wiley Blackwell,
Chichester. (Provides a definite depiction of plasmids and bacteriophages.)
Messing J (1983) New M13 vectors for cloning. Strategies in Enzymology 101: 20–78.
Sambrook J and Russell DW (2001) Molecular Cloning: A Laboratory Manual, third edn. Cold
Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
Willey, J., Sherwood, L. and Woolverton, C. (2007) Prescott's Microbiology, seventh edn.
McGraw Hill Higher Education, Maidenhead. (A great prologue to microbial science,
including plasmids and phages)
49
EMERGING TOOLS FOR CONSERVATION AND

MONITORING OF BIODIVERSITY
Sachin K. Shelake and Vishwas Y. Deshpande
Department of Zoology and Fisheries,
Yashwantrao Chavan Institute of Science, Satara, MS, India
Corresponding author E-mail: [email protected], [email protected]
Introduction:
The continual loss of biodiversity on Earth is a serious concern and challenge for the
twenty-first century, and there is worldwidepolitically aware agreement to limit or stop this loss.
The task is hampered in large part by a dearth of understanding about the status and
biodiversitydistribution– specifically because science has yet to define the vast majority of
species on the planet. To get credible estimates of population numbers and dispersal trends, all
conservation initiatives to conserve biodiversity rely on species and population monitoring.
Traditionally, visual surveys and individual counts have been used to physically identify species
for monitoring purposes.
Traditional monitoring procedures, on the other hand, continue to be troublesome due to
challenges in correctly identifying hidden species or life stages of juvenile, a steady reduction in
taxonomic knowledge, non-standardized sample and the intrusivenessof certain survey practices.
As a result, innovative and effective methodologies in order to keep track of biodiversity on a
wide scaleare urgently needed. DNA barcoding, environmental DNA (eDNA), community DNA,
invertebrate ingested DNA (iDNA) metabarcoding, and other developing conservation
technologies for monitoring biodiversity include DNA barcoding, environmental DNA (eDNA),
community DNA, and invertebrate ingested DNA (iDNA) metabarcoding.
DNA Barcoding:
DNA barcoding is a beneficial method foridentifying organisms at the molecular level.
This method entails amplifying a segment, the cytochrome oxidase enzyme I (COI) gene with
PCR, sequence analysis and matching it to a databank of already identified species. Various
genes, like as rbcL and matK, are frequently used in plants for barcoding identification. The aim
of this study was to find primers and reagents for polymerase chain reaction (PCR) DNA
barcoding on a wide range of taxonomic groups.
The published literature was evaluated for DNA barcoding methodologies and processes
for invertebrates, fish, mammals, and plants, and the PCR primers for these taxonomic groupings
were discovered and arranged. DNA barcoding methods were used to evaluate by researchers
50
(ISBN: 978-93-91768-45-4)
provided samples from a wide spectrum of species. Following the completion of the PCR
process, the PCR result was uploaded to the BOLD and NBCI databases for DNA sequencing
and analysis.DNA samples from fish, birds, animals, plants, and invertebrates might be
analysed.The findings of the analysis of samples supplied by Reclamation researchers were
given to the researchers. During the course of this assignment, we learnt a few things about DNA
barcoding. A literature review of DNA barcoding methods is included in this paper, as well as an
overview of the research and a discussion of the findings.
Taxonomic specialists have long used specialised language and literature to describe and
identify creatures; DNA-based identification methods, on the other hand, employ standardised
molecular biology procedures (DNA extraction, PCR, and DNA sequencing) to speed up the
identification of unknown organisms (Seifert et al., 2007). The objective of DNA barcoding
scientists is to develop a library of every life on the planet (Stoeckle et al., 2004).
Environmental DNA (eDNA):
Extracting DNA from environmental samples (environmental DNA - eDNA) to get
information on species, populations, and communities offers the potential to alleviate many of
the difficulties facing biodiversity monitoring (Baird and Hajibabaei, 2012; Kelly et al., 2014b).
The informationof DNA from higher species can be takenand examined in the environment has
been a major technical and scientific achievement in the recent decade. As creatures interact with
their surroundings, they continually discharge DNA into the environment.
This DNA can be derived from expelled cells or tissue in higher species, such as urine,
faeces, hairs and skin and, of course, from dead persons leaking genetic material. Although
macrobial eDNA possibly will reside largely inside mitochondria or tiny cells in some systems
(Turner et al., 2014), extracellular DNA will be present in the surroundings due to membrane
breakdown (Nielsen et al., 2007).
As a result, eDNA has been utilised to answer both practical and basic research problems
in a variety of fields, including ecology, molecular biology, environmental sciences and
palaeontology are some of the fields of study. The identification of various eDNA from macro-
organisms validated the method as actually important in a conservation context, and it has been
discovered in a wide range of ancient and modern terrestrial and aquatic habitat (Thomsen and
Willerslev, 2015).
Community DNA:
Since the commencement of high-throughput sequencing (HTS), the application of
metabarcoding as a technique of biodiversity identification has attracted biologists' interest.
(Margulies et al., 2005, Hajibabaei et al., 2011).Changes in sampling andvariations in research
laboratory processes, might affect later bioinformatics methods used to data processing and
51
confound the understanding of regional and of time biodiversity trends if there is no clarity
between these two source materials (Creer et al., 2016).
Typically, the selected populations are assembled in quantity (e.g., soil, malaise trap, or
net), and individuals are separated from other sample material and mixed together prior to bulk
DNA extraction (Creer et al., 2016, Lévêque et al., 2008). Macro-organism eDNA, on the other
hand, is extracted directly from an environmental substance (e.g., soil or water) without the need
to separate individual organisms or plant material from the sample, implying that the entire
organism is not there. Of course, community DNA samples may contain DNA from other
creatures' tissues, cells, and organelles (e.g., gut contents, cutaneous intracellular or extracellular
DNA). Similarly, macroorganism eDNA samples may catch complete tiny nontarget species
accidently (e.g., protists, bacteria). As a result, the distinction can, at least in part, be blurred in
reality (Deiner et al., 2014).
Metabarcoding of invertebrate ingested DNA (iDNA):
Using DNA metabarcoding to target vertebrate genetic material obtained from
invertebrates (such as leeches, mosquitoes, or ticks, among others). It might be considered a
unique example of eDNA metabarcoding because the DNA sources include swallowed stuff or
faeces.
Metabarcoding:
Metabarcoding is the process of barcoding DNA or RNA in such a way that many taxa
may be identified at the same time in the identical sample. The fundamental differenceamong
metabarcoding andbarcoding is, the metabarcoding is used to decide the species makeup of a
sample rather than focusing on a single organism. A barcode is made up of a brief variable gene
segment that is valuable for taxonomic classification and is bordered by gene with a high level of
conservation that may be utilised to design suitable primer (Pierre and colleagues, 2018).
Researchers at the University of Guelph came up with the concept of generic barcoding in 2003
(Hebert et al., 2003)
Metabarcoding, like ordinary barcoding, follows a step-by-step process that includes
DNA extraction; PCR amplification, sequencing, and data analysis are all steps in the process.
Different genes are used depending on whether the objective is to barcode a single species or to
metabarcode many species. In the second situation, a broader gene is used.Metabarcoding starts
with DNA/RNA from numerous distinct creatures taken from a single ambient or bulk sample,
rather than single species DNA/RNA.
References:
Baird, D.J., Hajibabaei, M., (2012). Biomonitoring 2.0: a new paradigm in ecosystem assessment
made possible by next-generation DNA sequencing. Mol. Ecol. 21, 2039–2044.
52
(ISBN: 978-93-91768-45-4)
Bello, F.; Chave, J.; Thuiller, W.; Wincker, P.; Cruaud, C.; Gavory, F.; Rasmussen, M.; Gilbert,
M. T. P.; Orlando, L.; Brochmann, C.; Willerslev, E.; Taberlet, P. (2012). DNA from soil
mirrors plant taxonomic and growth form diversity. Molecular Ecology. 21 (15): 3647–
3655. doi:10.1111/j.1365-294X.2012.05545.
Creer, Simon; Deiner, Kristy; Frey, Serita; Porazinska, Dorota; Taberlet, Pierre; Thomas, W.
Kelley; Potter, Caitlin; Bik, Holly M. (2016). The ecologist's field guide to sequence‐based
identification of biodiversity Methods in Ecology and Evolution. 7 (9): 1008–
1018. doi:10.1111/2041-210X.12574.
Dejean, T., Valentini, A., Duparc, A., Pellier-Cuit, S., Pompanon, F., Taberlet, P., Miaud, C.,
2011. Persistence of environmental DNA in freshwater ecosystems. PLoS ONE 6, e23398.
http://dx.doi.org/10.1371/journal.pone.0023398.
Hollingsworth PM, Graham SW, Little DP. (2011). Choosing and using a plant DNA barcode.
Plos One, 6 (5), e19254-e19254.
Deiner, Kristy; Altermatt, Florian (2014). Transport Distance of Invertebrate Environmental
DNA in a Natural River. doi:10.1371/journal.pone.0088786.
Deiner, Kristy; Bik, Holly M.; Mächler, Elvira; Seymour, Mathew; Lacoursière‐Roussel, Anaïs;
Altermatt, Florian; Creer, Simon; Bista, Iliana; Lodge, David M.; Vere, Natasha; Pfrender,
Michael E.; Bernatchez, Louis (2017). Environmental DNA metabarcoding: Transforming
how we survey animal and plant communities. Molecular Ecology. 26 (21): 5872–
5895. doi:10.1111/mec.14350.
Hebert, Paul D. N.; Cywinska, Alina; Ball, Shelley L.; Dewaard, Jeremy R. (2003). Biological
identifications through DNA barcodes Proceedings of the Royal Society of London. Series
B: Biological Sciences. 270 (1512): 313–321. doi:10.1098/rspb.2002.2218
Hajibabaei, Mehrdad; Shokralla, Shadi; Zhou, Xin; Singer, Gregory A. C.; Baird, Donald J.
(2011). Environmental Barcoding: A Next-Generation Sequencing Approach for
Biomonitoring Applications Using River Benthos. doi:10.1371/journal.pone.0017497.
Margulies, Marcel; et al. (2005). Genome sequencing in microfabricated high-density picolitre
reactors. Nature. 437 (7057): 376–
380. Bibcode:2005Natur.437..376M. doi:10.1038/nature03959. PMC 1464427. PMID 160
56220.
Merkes, Christopher M.; McCalla, S. Grace; Jensen, Nathan R.; Gaikowski, Mark P.; Amberg,
Jon J. (2014). Persistence of DNA in Carcasses, Slime and Avian Feces May Affect
Interpretation of Environmental DNA DataLOS ONE. 9 (11):
e113346. Bibcode:2014PLoSO...9k3346M. doi:10.1371/journal.pone.0113346.
53
Kerr KCR, Stoeckle MY, Dove CJ, Weigt LA, Frances CM, Herbert PDN. (2007).
Comprehensive DNA barcode coverage of North American birds. Molecular Ecology
Notes, 7, 535-543.
Lévêque, C., Oberdorff, T., Paugy, D., Stiassny, M. L. J., & Tedesco, P. A. (2008). Global
diversity of fish (Pisces) in freshwater. Hydrobiologia, 595(1), 545–567.
https://doi.org/10.1007/s10750-007-9034-0.
Nielsen, K.M., Johnsen, P.J., Bensasson, D., Daffonchio, D., 2007. Release and persistence of
extracellular DNA in the environment. Environ. Biosafety Res. 6, 37–53.
http://dx.doi.org/10.1051/ebr:2007031.
Pierre, Taberlet (2 February 2018). Environmental DNA : for biodiversity research and
monitoring. Bonin, Aurelie, 1979-. Oxford. ISBN 9780191079993. OCLC 1021883023.
Poinar, H.N., Hofreiter, M., Spaulding, W.G., Martin, P.S., Stankiewicz, B.A., Bland, H.,
Evershed, R.P., Possnert, G., Pääbo, S., 1998. Molecular coproscopy: dung and diet of the
extinct ground sloth nothrotheriops shastensis. Science 281, 402–406.
http://dx.doi.org/10.1126/science.281.5375.402.
Stoeckle M, Waggoner PE, Ausubel JS. (2004). Barcoding Life: Ten Reasons. Consortium for
the Barcode of Life, v3.0. Retrieved from:
http://www.barcodeoflife.org/content/barcoding-life-ten-reasons-pamphlet.
Seifert KA, Samson RA, Dewaard JR, Houbraken J, Levesque CA, Moncalvo JM, LouisSeize G,
Herbert PD. (2007). Prospects for fungus identification using COI DNA barcodes, with
Penicillium as a test case study. PNAS, 104(10),3901-3906.
Thomsen, P. F., & Willerslev, E. (2015). Environmental DNA - An emerging tool in
conservation for monitoring past and present biodiversity. Biological Conservation, 183,
4–18. https://doi.org/10.1016/j.biocon.2014.11.019.
Turner, C.R., Barnes, M.A., Xu, C.C.Y., Jones, S.E., Jerde, C.L., Lodge, D.M., 2014. Particle
size distribution and optimal capture of aqueous macrobial eDNA. bioRxiv.
http://dx.doi.org/10.1101/001941.
Valiere, N., Taberlet, P., 2000. Urine collected in the field as a source of DNA for species and
individual identification. Mol. Ecol. 9, 2150–2152. http://dx.doi.org/ 10.1046/j.1365-
294X.2000.11142.x.
Willerslev, E., Cooper, A., 2005. Review paper. Ancient DNA. Proc. R. Soc. B Biol. Sci. 272, 3–
16. http://dx.doi.org/10.1098/rspb.2004.2813.
54
(ISBN: 978-93-91768-45-4)
DIMETHOATE: KILLER PESTICIDE

Chandani B. Jamdade
S. G. M. College, Karad,
RIRD, Satara, Maharashtra, India
The pesticides are one of the most potentially harmful chemicals liberated in the
environment in an unplanned manner. Pesticides are biologically active chemical. The misused
of pesticide may be harmful to humans, animals and environment. Dimethoate is widely used as
a potent pesticide and acaricide in many countries and has been shown to produce some adverse
health effects. Dimethoate, (IUPAC name O, O-dimethyl SN- methyl carbamoyl methyl
phosphorodithioate) is widely used against a broad range of insects and mites and is also used for
indoor control of houseflies. WHO and International Program on Chemical Safety (IPCS) said
that the extensive use of Dimethoate poses a health hazard to animals and humans because of its
persistence in water, soil and crops. Dimethoate classified as moderately hazardous by WHO and
the oral dose of dimethoate is biotransformed to its oxygen analogue Omethoate, which is the
active form, by hydrolysis of the methyl ester group and removal of the methyl-amido group.
Omethote is considerably more toxic than dimethoate. U. S. Environmental Protection Agency
(USEPA) has registered Dimethoate a systemic organophosphate insecticide but interim in 2006
it released reregistration eligibility decision (IRED) document for dimethoate in accordance with
FQPA requirements.
A great proportion of acute poisoning cases are caused by exposure to pesticides,
especially organophosphate (OP) compounds. Dimethoate is rapidly absorbed from the
gastrointestinal tract of the body and its poisoning is usually block neuromuscular transmission
in both animals and humans. The primary mechanism of action of OP pesticides is based on
inhibition of the acetylcholinesterase (ache) enzyme. Once ache has been inactivated;
acetylcholine (ach) accumulates throughout the nervous system, resulting in overstimulation of
muscarinic and nicotinic receptors. Signs and symptoms of op poisoning can be divided into
three broad categories, as muscarinic, nicotinic, and central nervous system effects.
The toxicity of organophosphorus insecticides results in negative effects on many organs
and systems such as the liver, kidney, nervous system, immune system and reproductive system
55
in many animals. Furthermore, organophosphorus insecticides exert their biological effects

through electrophilic attack on the cellular constituents of hepatic and brain tissues with
simultaneous generation of reactive oxygen species. Dimethoate induces a number of
morphological and functional changes in several tissues and organs of the body and it displays
the ability to induce free-radical processes. Some study indicates that administration of
dimethoate led to induction of oxidative stress and which was affect on increased concentration
of MDA in liver. Toxicity of organophosporus insecticides used compounds against human and
animals were always evaluated by assessment of such biochemical parameters alterations and
histopathological changes in tissues and organs. Kidney is one of the targets organs of
experimental animals attacked by organophosphorous compounds.
Management of severe poisoning is difficult, requiring intensive care and use of atropine
and oxime cholinesterase reactivators. The acute and chronic toxicity of dimethoate is
moderately high by ingestion, inhalation and dermal absorbtion. By considering all these
problems this pesticide is banned in some countries. Though it is banned, is used on field as well
as in house for controlling variety of pest. So for our safety purpose we should stop the use of
this hazardous pesticide for healthy life.
References:
https://www.who.int/water_sanitation_health/water-quality/guidelines/chemicals/dimethoate.pdf
https://archive.epa.gov/pesticides/reregistration/web/pdf/revised_dimethoate_ired.pdf
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2493390/
https://pubmed.ncbi.nlm.nih.gov/22840586/
56
(ISBN: 978-93-91768-45-4)
DIVERSITY OF MOTHS (LEPIDOPTERA: HETEROCERA) AND THEIR

POTENTIAL ROLE AS A CONSERVATION TOOL IN PROTECTED AREAS OF
KATEPURNA WILDLIFE SANCTUARY OF AKOLA (M.S.)
Priyanka M. Ramteke
Shri Shivaji Art, Commerce and Science College, Akola, M. S.
Abstract:
Moths are diverse group of insects belonging to the order Lepidoptera and regarded as
one of the indicators of a healthy environment. Despite a large number of studies been taken up
on the documentation of various wildlife taxa found in and around Katepurna wildlife Sanctuary
of Akola by the biologists, information on Moths of this region remains unknown. The present
study is the first documentation on the moth species of Katepurna wildlife Sanctury of Akola.
The study will be carried out throughout the year surveying areas mostly in the protected areas of
Katepurna wildlife Sanctuary of Akola (M.S.) and agricultural lands. Light trapping equipped
with a 18w UV-Actinic tube attached to a white sheet 6 x 4 feet joined to two poles and then the
tube powered by 6v Battery, which will be used to record moths from playgrounds of college,
human settlements and agricultural lands. In total, 12 moth individuals were recorded belonging
to 12 species Within 12 genera falling under 8 families.. The family Crambidae -3 represented
the highest number of species, followed by Geometridae -2 , Saturniidae -2. . The less commonly
observed species belonging to families are Pyralidae-1, Uraniidae-1, Noctuidae-1, Erebidae -1
and Sphingidae-1 individuals respectively.
Keywords: Moths, Lepidoptera, Katepurna Wildlife Sanctuary
Intoduction:
Akola town of the Vidharbha State of Maharashtra situated at latitude 20.70 North and
longitudinal 77.07 0 East.It is an altitude of 925ft (287m) to 1036.745ft (316m) above sea level.
Akola has tropical savanna climate Annual temperature range from a high of 47.60c to a low
2.20c. Annual rain fall averages 800mm. Katepurna Sanctuary is situated in the district of Akola,
which comes under the Vidarbha region. The sanctuary occupies a large part of catchment area
57
of the Katepurna reservoir. Due to the presence of large amount of water, birds get attracted
towards the place. The period between October to June is the ideal time for visiting the place.
Flora- The place is very rich in vegetation cover and mainly comprises of southern tropical
deciduous forests. There are more than 115 species of plants & trees like Bahada, Aola, Tendu,
Dhawada, Salai, Moha, Teude and many more. Fauna- The sanctuary is a home to a wide array
of animals like Black buck, Hyena, Wolf, Nilgai, Leopard, Jungle cat, Hare, Wild boar, Monkey
etc. The chief attraction of the sanctuary is the Barking deer and the Four-horned Antelope.
One can also see various species of birds like Peafowl, eagles, woodpecker etc.
Katepurna Wildlife Sanctuary is spread over 73.69 sq. km. It covers a great part of the catchment
area of 12 the Katepurna reservoir. It is well known for for-horned antelope and barking deer.
Among the winged insects, the moths belong to the scientific order Lepidoptera including the
butterflies. They can be distinguished from all other insects by the two pair of wings and the
body that is scale covered. Regarded as indicators of healthy environment, inventory of
Lepidoptera specially the overlooked group is the first step to know what are the species present
in an area and it is essential to ensure future taxonomical and ecological studies of these taxa and
implement conservation perspectives for moth individuals as well as their associated habitats.
Global estimates show that there are 1, 27,000 species of moths distributed over the
world and of which, 12,000 species are reported from India. Moths are in general are least
studied taxa across the globe and in India.
Despite a large number of studies been taken up on the documentation of various wildlife
taxa found in and around Katepurna wildlife Sanctury of Akola by the biologists, information on
Moths of this region remains unknown. The present study is the first documentation on the moth
species of Katepurna wildlife Sanctury of Akola.
Materials and Method:

The study will be carried out throughout the year surveying areas mostly in the protected
areas of Katepurna wildlife Sanctuary of Akola (M.S.) and agricultural lands. Light trapping
equipped with a 18w UV-Actinic tube attached to a white sheet 6 x 4 feet joined to two poles and
then the tube powered by 6v Battery, which will be used to record moths from playgrounds of
college, human settlements and agricultural lands.
The moths will be photographed and identified and those that will be difficult to identify
will be kept for proper identification. Among the literatures, Fauna of British India: Moths
Volume I-V by G.F Hampson will be referred for identification along with other journals.
58
(ISBN: 978-93-91768-45-4)
Results and Discussion:

A total of 12 moth individuals were recorded belonging to 12 species Within 12 genera
falling under 8 families. A checklist of the moth Species is tabulated in Table 1.
Table 1: List of moth species of protected areas of Katepurna wildlife Sanctuary, Akola
Sr. No. Name of the species Family Genus

1. Automeris bilinea Saturniidae Antheraea
2. Plodia interpunctella Pyralidae Plodia
3. Hymenia recurvalis Crambidae Hymenia
4. Pseudopanthera macularia Geometridae Pseudopanthera
5. Micronia aculeata Uraniidae Micronia
6. Spirama retorta Noctuidae Spirama
7. Diaphania indica Crambidae Diaphania
8. Thalassodes immasaria Geometridae Thalassodes
9. Antheraea mylitta Saturniidae Antheraea
10. Pygospila tyres Crambidae Pygospila
11. Asota planaria Erebidae Asota
12. Agrius convolvuli Sphingidae Agrius
The accumulation of moths in a light source depends on the type of light source, plant
communities occurring around the study site, temperature, weather conditions, altitudinal
gradient, and the type of methods implemented. The moths recorded by visiting the mentioned
Localities and sheet light trap method was a valuable source for developing a preliminary
data record for moths occurring in this region. The notable species accumulated in the Katepurna
wildlife habitats were Automeris bilinea and Antheraea mylitta belongs to Saturniidae family on
their host plants namely Syzygium cumini (jambul), Hardwickia binata (anjan).
Plodia interpunctella belong to family Pyralidae. It feeds on a wide variety of dry
vegetable matter, including grains, nuts, dry beans, dried fruits, dried flowers, bird seed, dry pet
foods, processed foods such as cereals and crackers, and confections.
Hymenia recurvalis, Diaphania indica and Pygospila tyres belong to family Crambidae
species of observed as a minor pest of potato and cucumber near the crop land area of Katepurna.
59
Geometridae family represents two species of moth Pseudopanthera macularia and Thalassodes
immasaria on their host plant namely, Ricinus communis, Mangifera indica (mango).
Images of some moths of protected areas of Katepurna wildlife Sanctuary of Akola
Plodia interpunctella Indian Beet webworm moth

Automeris bilinea
meal moth Hymenia recurvalis
Pseudopanthera macularia Micronia aculeate Spirama retorta
Daphnia indica Thalassodes immissaria Antheraea mylitta
Pygospila tyres Asota planaria Agrius convolvuli
60
(ISBN: 978-93-91768-45-4)
Micronia aculeata belongs to family Uraniidae observed near their host plant namely
syzygium jambo.
The species Spirama retorta , Asota planaria and Agrius convolvuli represented the
family Noctuidae, Erebidae and Sphingidae respectively These three families appeared to be
scarce representing single species. Near the host plant namely Solanum lycopersicum
(Tomatoes), Ficus plant and on hibiscus flower respectively.
Conclusion:
Moths also play a vital role in telling us about the health of our environment, like the
canary in the coalmine .since they are so widespread and found in so many different habitats, and
are so sensitive to changes; moths are particularly useful as indicator species. Monitoring their
number and ranges can give us vital clues to changes in our own environment, such as the effect
of new farming practices, pesticides, air pollution and climate change.
References:
Barsagad D. D., et al., (2012): Vanya Tasar Silkworm, Antheraea Mylitta Eco-Race Bhandara,
The Local Race And Its Conservation Strategy (Lepidoptera: Saturniidae) Journal Or
Science Information / Special Issue – 3.
Beljaev E. A. (1997): A New Subspecies Of The Pseudopanthera Macularia L. (Lepidoptera:
Geometridae, Ennominae) From South Siberia Number 51: 1-7.
Clerk (1764): Spirama retorta Icon.Insect. rariorum2: pl. [54]
Cramer (1780): Pygospila tyres Cramer P. De uitlandsche Kapellen voorkomende in de drie
waereld-deelen Asia, Africa en America, by een verzameld en beschreeven door den heer
Pieter Cramer (etc.). 1779–1780.
Fabricius 1775: Asota caricae ICAR national bureau of agricultural insect resources. Retrieved
22 july, 2016.
Guenee in Boisduval and Guenee (1857): Micronia aculeate Histoire Naturelle des Insectes.
Species General des Lepidopteres. Volume 10.
Hongxiang Han and Dayong Xue (2011): Thalassodes And Related Taxa Of Emerald Moths In
China (Geometridae, Geometrinae) Zootaxa 3019: 26–50
Jeyasankara A., and D. Gokilamania (2016) Biology and eco-friendly control of Amaranths
pests, Hymenia recurvalis Fabricius and Psara basalis Walker (Lepidoptera: Crambidae)
International Journal of Academic Studies 2(4): 218-230.
61
Linnaeus (1758): Agricus convovuli Convovulus hawkmoth Sphingidae of Eastern palaeartic

retrieved December 13, 2018.
Lranipour Shahzad and Azadeh Farzamand (2011): Life History Of Indian Meal Moth, Plodia
Interpunctella (Hubner) In Laboratory Global Conference on Entomology, March 5-9,
2011 Chiant Mai, Thailand.
Saunder (1851): Diaphania indica Peter, C and David, B; Insect –host plant interaction in
relation to development of Daphnia indica. The journal of Bombay Natural History
Society., 89, 1992.
62
(ISBN: 978-93-91768-45-4)
CLIMATE CHANGE AND ITS INTERACTIONS WITH PEOPLE AND

ECOSYSTEMS
Ansari Abdullah Daniyal
Department of Geography,
MGV’s Arts, Commerce and Science College Malegaon City,
Dist. Nashik, Maharashtra, India
The quick anthropogenic environmental change that is being knowledgeable about the
mid twenty-first century is personally weaved with the wellbeing and working of the biosphere.
Environmental change is affecting biological systems through changes in mean conditions and in
environment inconstancy, combined with other related changes like expanded sea fermentation
and air carbon dioxide fixations. It likewise collaborates with different tensions on environments,
including debasement, defaunation and discontinuity. There is a need to comprehend the
biological elements of these environment impacts, to recognize focal points of weakness and
flexibility and to distinguish the board intercessions that might help biosphere strength to
environmental change.
Simultaneously, biological systems can likewise aid the relief of, and transformation too,
environmental change. The instruments, potential and cutoff points of such nature-based answers
for environmental change should be investigated and measured. This paper acquaints a topical
issue committed with the cooperation between environmental change and the biosphere. It
investigates novel points of view on how biological systems react to environmental change, how
biological system flexibility can be upgraded and how environments can help with tending to the
challenge of an evolving environment. It draws on a Regal Society-Public Foundation of
Sciences Discussion held in Washington DC in November 2018, where these subjects and issues
were examined. We close by distinguishing a few needs for scholastic examination and
pragmatic execution, in request to boost the potential for keeping a different, tough and well-
working biosphere under the difficult states of the twenty-first century.
This article is essential for the topic issue 'Environmental change and biological systems:
dangers, openings and arrangements'.
The drawn out great soundness of populaces relies upon the proceeded with security and
working of the biosphere's biological and actual frameworks, frequently alluded to as life-
emotionally supportive networks. We overlook this since quite a while ago settled authentic truth
63
at ourrisk: yet it is all around not entirely obvious this reliance, especially at a time at the point
when the human species is turning out to be progressively urbanized and removed from these
regular frameworks. The world's environment framework is a vital piece of this complex of life-
supporting cycles, one of numerous enormous regular frameworks that are presently going under
strain from the expanding weight of human numbers and monetary exercises. By coincidentally
expanding the centralization of energy-catching gases in the lower environment, human activities
have started to enhance Earth's regular nursery impact. The essential test confronting the world
local area is to accomplish adequate decrease in ozone harming substance outflows to stay away
from hazardous impedance in the environment framework. Public state run administrations,
through the UN System Show on Environmental Change (UNFCC), are submitted on a basic
level to looking forthis result. By and by, it is demonstrating hard to track down a politically OK
strategy—frequently in view of anxieties about conceivable present moment financial outcomes.
This volume looks to depict the specific situation and interaction of worldwide
environmental change, its genuine or possible effects on wellbeing, and how human social orders
ought to react, by means of both variation techniques to diminish effects and aggregate activity
to decrease ozone harming substance outflows. As displayed later, a significant part of the
resultant danger to human populaces and the biological systems whereupon they depend comes
from the extended incredibly fast pace of progress in climatic conditions. For sure, the
possibilityof such change has animated a lot of new logical exploration over the past decade,
quite a bit of which is clarifying the complex natural aggravations that can affect the human
prosperity and wellbeing—as in the accompanying model. The US Worldwide Change
Exploration Program (Gold country Territorial Evaluation Gathering) as of late archived how the
different impacts of environmental change on sea-going environments can interface and echo
through trophic levels in flighty ways.For instance, warming in the Cold district has diminished
how much ocean ice, impeding endurance rates for walrus and seal little guys that go through
piece of their time on earth cycle on the ice. With less seal little guys, ocean otters have turned
into the elective food hotspot for whales. Ocean otters feed on ocean imps, and with less ocean
otter‘s ocean imp populaces are growing and burning-through a greater amount of the kelp that
gives favorable places for fish. Less fish fuel the decreases in walrus and seal populaces. By and
large, there is less food accessible for the Yupik Eskimos of the Icy who depend on these
species.
Worldwide environmental change is consequently a critical expansion to the range of
ecological wellbeing dangers looked by mankind. The worldwide scale makes for newness—
albeit a large portion of its wellbeing impacts involve increments (or diminishes) in natural
impacts of climatic minor departure from human science and wellbeing. Conventional ecological
64
(ISBN: 978-93-91768-45-4)
wellbeing concerns long have been centered on toxicological or microbiological dangers to

wellbeing from neighborhood natural openings. Notwithstanding, in the early long periods of the
twenty-first century, as the expanding human effect on the climate keeps on adjusting the planet's
land, organic and biological frameworks, a scope of bigger scope ecological risks to human
wellbeing has arisen. Notwithstanding worldwide environmental change, these include: the
wellbeing chances presented by stratospheric ozone consumption; loss of biodiversity; weights
on earthbound what's more sea food-delivering frameworks; changes in hydrological frameworks
and the provisions of freshwater; and the worldwide scattering of steady natural toxins.
Environmental change and stratospheric ozone exhaustion are the most popular of these
different worldwide ecological changes. Human social orders, nonetheless, have had long
experience of the changes of environment: climatic cycles have left incredible engravings what's
more scars on the historical backdrop of humanity. Civilizations like those of antiquated Egypt,
Mesopotamia, the Mayans, and the Vikings in Greenland and European populaces during the
four centuries of Little Ice Age, all have both benefited and experienced nature's incredible
climatic cycles. Recorded examinations likewise uncover broad fiascos, social interruption and
infection flare-ups in light of the more intense, between yearly, semi intermittent ENSO (El Niño
Southern Wavering) cycles. The exhaustion of soil fruitfulness and freshwater supplies, and the
blunder of water catchment bowls by means of exorbitant deforestation, likewise has added to
the decay of different local populaces throughout the long term.
Today, environment researchers foresee that mankind's expanding emanation of ozone
harming substances will initiate a drawn out change on the planet's environment. These gases
contain, mainly, carbon dioxide (generally from petroleum product burning also timberland
consuming), in addition to different other hotness catching gases like methane (from flooded
horticulture, creature cultivation and oil extraction), nitrous oxide furthermore different human-
made halocarbons. Without a doubt, most environment researchers now think that the amassing
of these gases in the lower environment has added to the solid ongoing upswing in world normal
temperature. In its Third Appraisal Report, distributed in 2001, the Intergovernmental Board on
Environmental Change (IPCC) expressed: "There is new and more grounded proof that the
majority of the warming saw throughout the most recent 50 years is inferable from human
exercises"
Environment and human wellbeing: an old battle:
Whoever wishes to research medication appropriately, ought to continue accordingly: in
the first spot to think about the periods of the year, and what impacts every one of them
65
produces, for they are not all indistinguishable, but rather vary much from themselves as to their
changes.
Acknowledgment that human wellbeing can be impacted by a wide scope of biological
disturbances, resulting upon environmental change, is a new turn of events, reflecting the
expansiveness and refinement of present day logical information. By and by, the easier thought
that human wellbeing and sickness are connected to environment presumably originates before
recorded history. The Greek doctor Hippocrates (around 400 BC) related pandemics to
occasional climate changes, composing that doctors ought to have "due respect to the seasons of
the year, and the illnesses which they produce, and to the conditions of the breeze impossible to
miss to every nation and the characteristics of its waters". He admonishes themto observe "the
waters which individuals use, regardless of whether they be muddy and delicate, or then again
hard and running from raised and rough circumstances, and afterward if saltish and ill suited for
cooking," and to notice "the areas of towns, and of the encompassing country, regardless of
whether they are low or high, hot or cool, wet or dry and of thediet and routine of the occupants".
Potential wellbeing effects of environmental change:
Worldwide environmental change would influence human wellbeing through pathways of
shifting intricacy, scale and unequivocal quality and with various planning. Also, effects would
shift topographically as a capacity both of climate and geology and of the weakness of the nearby
populace. Effects would be both positive and negative (albeit master logical surveys expect
dominatingly negative). This is nothing unexpected since climatic change would disturb or in
any case modify an enormous reach of regular natural and actual frameworks that are a necessary
piece of Earth's life-emotionally supportive network. By means of environmental change people
are adding to a change in the states of life on the planet.
Population vulnerability and adaptive responses:
Human populaces, similarly as with people, fluctuate in their weakness to specific
wellbeing results. A populace's weakness is a joint capacity of, first, the degree to which a
specific wellbeing result is delicate to environmental change and, second, and the populace's
ability to adjust to new climatic conditions. The weakness of a populace relies upon variables,
for example, populace thickness, level of financial turn of events, food accessibility, pay level
and dissemination, nearby ecological conditions, previous wellbeing status and the quality and
accessibility of general medical services.
Transformation alludes to activities taken to diminish the effect of (expected)
environmental change. There is a progressive system of control procedures that can assist with
securing populace wellbeing. These systems are classified as: regulatory or administrative,
designing, individual conduct. Authoritative or administrative move can be made by government,
66
(ISBN: 978-93-91768-45-4)
requiring consistence by all or assigned classes of people. Then again, versatile activity might be
empowered on a willful premise, through backing, training or monetary motivating forces. The
previous sort of move would ordinarily be made at a supranational, public or local area level; the
last option would go from supranational to individual levels. Variation techniques will be either
receptive, in light of environment impacts, or expectant, in order to reduce vulnerability.
Conclusions:
Over the ages human social orders have debased or changed nearby environments and
altered provincial environments. Unprecedented, the total human effect currently has
accomplished a worldwide scale, mirroring the new fast expansion in populace size and energy-
serious, high-throughput, mass utilization. The total populace is experiencing new human-incited
changes in the lower and center airs and overall consumption of different other regular
frameworks (for example soil richness, springs, sea fisheries and biodiversity overall).
Regardless of early acknowledgment that such changes would influence monetary exercises,
foundation and oversaw biological systems, there has been less mindfulness that such huge scope
natural change would debilitate the backings for solid life. Luckily that is presently starting to
change. For sure, this volume looks to introduce a far reaching conversation of the connection
between worldwide environmental change and human populace wellbeing.
Worldwide environmental change is probably going to change the recurrence of
outrageous climate occasions: typhoons might increment as ocean surface waters warm; floods
might increment as the hydrological cycle heightens; and heat waves might increment in mid-
mainland areas. As talked about exhaustively in later parts, an adjustment of the recurrence and
force of hotness waves and cold spells would influence occasional examples of dreariness and
mortality. The creation of different air toxins and of allergenic spores and dusts would be
impacted by hotter and wetter conditions. Environmental change likewise is relied upon to
influence wellbeing by means of different roundabout pathways, including the examples of
irresistible illnesses; the yield of food-delivering frameworks ashore and adrift; the accessibility
of freshwater; and, by adding to biodiversity misfortune, may undermine and debilitate the
biological system administrations whereupon human culture depends.
Variations to the wellbeing risk presented by worldwide environmental change can be
both proactive and receptive, and can happen at the full scale, meso and small sizes; that is, at the
populace, local area and individual levels. Environmental change addresses an oddball
worldwide test so there will be restricted freedom to complete fundamental assessment of
transformation choices. There is in this manner a solid case for reasonability, both in moderating
67
environmental change and in adjusting to its effects. This subject is probably going to turn into a
significant topic in populace wellbeing research, social strategy improvement and support during
this first decade of the twenty first century. Without a doubt, thought of worldwide climatic-
ecological risks to human populace wellbeing will assume a focal part in the supportability
progress banter.
References:
Abu al-Fazl (1873-94): Ain-I Akbari, trans. H. Blochmann, H. S. Jarrett, 3 vols. (Asiatic Culture
of Bengal):
Aftab, E. furthermore G. M. Hickey (2010): Woodland Organization Difficulties in Pakistan:
The Instance of the Patriata Held Timberland and the New Murree Advancement, Global
Ranger service Survey, 12/l :97-105.
Agarwal, Bina (2010): Sexual orientation and Green Administration: The Political Economy of
Ladies Quality Inside and Past People group Ranger service (Oxford College Press):
Agnihotri, Indu (1996): Environment, Land Use and Colonization: The Channel Settlements of
Punjab, Indian Financial and Social History Audit, 33/l :37-58.
Agrawal, Arun (2005): Environmentality: Advances of Government and the Creation of Subjects
(Duke College Press):
Fagan, B. (1999): Floods, starvations and rulers. El Niño and the fate of civilisations. New York,
USA, Fundamental Books.
Folke, C. et al. (1996): imitless resource arrangement. In: Getting viable. Costanza, R. whats
more Segura, O. eds. Washington, DC, USA, Island Press,
Intergovernmental Board on Ecological Change (IPCC): Ecological Change 2001: Third
Assessment Report (Volume I): Cambridge, UK Cambridge School Press, 2001.
McMichael, A.J. (2001): Human unsettled areas, conditions and ailment. Cambridge, UK,
Cambridge School Press,
Vitousek, P.M. et al. (1997): Human authority of Earths organic frameworks. Science 277: 494–
499 Wackernagel, M. et al. (2002): Following the organic overshoot of the human
economy. Strategies of the public Organization of Sciences, 99:9266–9271
World Bank. World progression report. Progression and the environment. Oxford, UK, Oxford
School Press, 1992.
68
(ISBN: 978-93-91768-45-4)
SERICULTURE BASED INTEGRATED FARMING SYSTEM FOR

SUSTAINABLE AGRICULTURAL DEVELOPMENT
Suraksha Chanotra* and Muzafar Ahmad Bhat
P.G. Department of Sericulture,
Poonch Campus, University of Jammu, Jammu & Kashmir (India) 185 101
*Corresponding author E-mail: [email protected]
Abstract:
Sericulture being an efficient agro-enterprise offers appropriate combination and
utilization of wide range of farm industries including, horticulture, livestock, fishery, forestry
and poultry etc. Integrated use of these sectors in a judicious manner ensures the means to raise
the farmer‘s productivity, profitability and helps to elevate their economic status as well.
Integrated farming system in combination with sericulture interacts adequately with environment
without dislocating the ecological balance on one hand and attempts to meet the national goals of
increased production on the other. IFS is a strategy to ensure sustainable use of the natural
resources for the benefit of present era while ensuring sufficient stock for future generations. In
this direction, sericulture industry offers the best suited integrated and efficient disposal of
resources specifically for the rural and marginal farmers.
Keywords: Sericulture, agro-enterprise, integrated, socio-economic, sustainable, resources
Introduction:
Indian economy is predominantly constituted by rural and agriculture oriented marginal
and small scale farmers constituting approximately 76.2 per cent of farming community of the
country (Kumar et al., 2012). In India agriculture remains the prime occupation of marginal and
rural farmers and limited availability of resources with confined seasonal climates prevailing in
the country have made the farmers to look for supporting secondary sectors such as sericulture
(Rai, 2006). Significant use of silkworm in agriculture and industrial sector has promoted the
importance of sericulture in all over the world (Ramesh- Babu et al., 2009) and it also
contributed in strengthening the national economy by generating employment on a large scale
(Chanotra et al., 2019). Integrated Farming System or IFS aims at the overall development of
agriculture and allied sectors by ensuring efficient, effective and judicious use of available
resources following a well planned management strategy for sustainable agricultural production
69
and enhancing the economic development so as to meet the current desires and preserving the
sufficient resources and basic stock material for future generations with main emphasis on
maintaining the quality of environment (Yadav et al., 2019). As sericulture is a rural based
cottage industry involving various labour intensive sectors like moriculture, silkworm rearing
and post-cocoon sector or industrial sector, integrated farming system in combination with
sericulture can be viewed as in combination with various enterprises in which the end-products
and of one sector can be utilized as the raw material for the next sector. For example, the waste
of silkworm rearing including excreta, urine contaminated leaves, undersized worms etc. can be
used for preparation of high quality FYM. The farm yard manure obtained after recycling the
sericultural waste will serve as an excellent biofertilizer especially for vegetables, flower
nurseries and other agricultural crops etc. The mulberry twigs obtained from pruning or training
or from shoot rearing can be used as fodder for cattles. Sericultural farmers can also adopt
floriculture, horticulture and fisheries etc. as part time enterprise as mulberry field allows
intercropping with other horticultural and floricultural crops. Moreover, the by-products
generated in post cocoon or industrial sector can also be utilized as raw material for poultry and
fisheries. For example, the left over pupae after reeling of cocoons can be easily processed to be
used as potential dietary material for poultry and fisheries. Thus, different enterprises under
sericulture industry shares high degree of correlation with one and other and one can yield great
benefit by adopting sericulture as an integrated agro-enterprise.
Criteria for integration of various agri-enterprises (Rana and Rana, 2011):
Integration of different agri-enterprises depends on some basic and flexible criteria which
include:
1. Soil health and climatic conditions prevailing in a particular area.
2. Availability of required resources including land and labour to work with.
3. Current status of available resources.
4. Expected economic output of the proposed integrated farming system.
5. Management strategy and skills of the famers involved.
Scope of Integrated Farming System:
Integrated farming system comprises of collective development of moriculture,
sericulture, other agricultural crops, dairy or livestock, poultry and horticulture etc. for
sustainable development and prosperity of farmers involved in their respective sectors.
Considering the fact that integrated implementation of various agri-enterprises in a well planned
strategy with optimum efforts yields better results than any of the enterprise alone. Therefore,
sericulture is the only enterprise that can be mingled easy with any of the others and offers
70
(ISBN: 978-93-91768-45-4)
tremendous scope and opportunities to the farmers to elevate their socio-economic status by
generating additional income.
Sericulture and Horticulture:
In India Horticulture is a blooming industry and holds unique importance among various
fields of agriculture as the fruits and vegetables cultivated under horticulture are consumed as
basic commodity by all being rich in nutritive values (Kumar et al., 2012). Horticulture offers
excellent avenues for generating employment and contributes in national economy by earning
foreign exchange. The production can be increased by increasing the area under cultivation and
adopting the idea of mixed cropping. Mulberry the host plant of silkworm Bombyx mori can be
planted as tall trees under mixed cropping system or on side bunds of the vegetable crop fields.
Among various horticulture crops, mango, coconut and sapota can be easy integrated with
mulberry cultivation (Kerutagi et al., 2019). Thus the farmers need not to engage their entire land
for mulberry cultivation and can generate additional income by practising sericulture on part
time basis. Moreover the concept of byproduct utilization works well in this direction as the
waste generated during silkworm rearing including silkworm litter, undersized and dead worms,
left over leaves and twigs etc. can be recycled to produce organic manure when mixed with
residual wastes of other agricultural crop, which can serve as an excellent biofertilizer to boost
the growth and nutritive value of vegetables and fruits thus minimizing the dependency on
chemical fertilizers.
Sericulture and floriculture:
Flower cultivation as a supplementary activity helps to improve the economic
opportunities for the rural farmers. They can adopt floriculture by starting small nurseries with
limited space and time. Sericultural wastes recycled into biofertilizers can be used to promote the
growth of flower crops. Moreover, cultivation of flowers as intercrop in mulberry fields can also
be adopted for additional benefits (Baishya et al., 2004). Cultivation of flowers like marigold and
gladiolus or cut flowers can be easily incorporated in IFS (Kumar et al., 2012).
Sericulture and Dairy:
Through a well planned management strategy various small enterprises like sericulture,
moriculture and dairy farming can be integrated which would increases involvement of labour
folk and offers opportunities to the farmers to gain additional benefits. In an experiment
conducted by Nagaraju and Raghavendra (2016), it has been reported that adoption of integrated
farming particularly in combination of crop, dairy, sheep and sericulture generated the most
71
beneficial results to the farmers in terms of both income and employment generation which was
recorded as 322 man days per year.
Sericulture and Poultry:
In India sericulture is viewed as a subsidiary occupation by the rural farmers. Moreover
the seasonal fluctuations compel the farmers to view other alternatives. Broiler or poultry
industry provides a good source of income to the marginal farmers. Silkworm pupae are
considered as an excellent source of protein, fats, essential vitamins and minerals (Longvah et
al., 2011). Thus, mixture of silkworm larvae and pupae blended in appropriate proportions with
other food materials can be served as an excellent feed to the poultry fouls. Thus sericulture in
combination with poultry can be viewed as one of the promising technology in augmenting
handsome income for the farmers as this combination opens avenues for small scale and
commercial agriculture business and hence this type of diversification yields fruitful results (M.
Prein, 2002). On the other hand waste generated in poultry can be recycled to produce organic
manure that can be applied as biofertilizer to boost the growth of mulberry plants which in long
run helps the worms to produce good cocoons thus boosting the farmer‘s resources and income
(Kumar et al., 2012, Aklilu Nigussie, 2018).
Sericulture and vermicomposting:
Vermicomposting is the process of conversion or breakdown of crop residues of
agricultural origin into organic manure with the addition of biological agents known as vermicast
or earthworms. Various end products are generated at each and every stage of sericulture which
can be utilized by vermicomposting and the vermicompost thus produced can be used for
cultivation of various agricultural crops. In a study conducted by Kerutagi et al., 2019, it has
been reported that the average income of farmers increased by 38.31 per cent by adopting
integrated farming system involving vermicomposting as an integral enterprise.
Some additional integrated programmes involving sericulture and other allied agri-enterprises
laid emphasis on the following aspects of IFS and sericulture (Baishya et al., 2004):
1. Mushroom production by utilizing seri-wastes as straw and compost.
2. Intercultivation of non-mulberry host plants like soam and salu and promotion of Vanya
sericulture.
3. Integration of sericulture and pig industry.
4. Integration of sericulture and fishery.
5. Integration of sericulture and apiculture.
72
(ISBN: 978-93-91768-45-4)
Interactions and linkages:

Various agro-enterprises integrated under IFS can be viewed as an inter-related series or
chain of activities as given below:
Dairy or livestock farming Apiculture Horticulture and Vegetable production
Integrated Farming System
Floriculture Vermicomposting Poultry farming
Figure 1: Various agri-enterprises involved under IFS
Advantages of Integrated Farming System:

Integrated farming system offers numerous advantages as it ensures adequate pooling and
utilization of available natural resources in an efficient manner so as to provide tremendous
opportunities to the farmers to gain more and more profit in terms of productivity and income
generation. Some important points under this category are described below:
1. Enhanced production:
Integrated farming system if adopted and executed through a proper management
strategy, results in tremendous increase in terms of production per unit area at given interval of
time.
2. Increased profitability:
Integrated farming system provides great profit in terms of income generated as the
integration of various enterprises ensures minimum input and maximum output returns i.e.
output/input ratio remains high in IFS.
3. Sustainability:
As IFS ensures integrated and judicious use of available resources it also ensures the
sustainability for future too.
4. Eco-friendly:
Integrated farming system aims at efficient utilization of natural resources while
maintaining the ecological balance.
73
5. By-product utilization:
Integrated farming system ensures zero or minimum wastage of any material. Maximum
emphasis is made on utilization of waste products and mostly the inter-related fields works in a
chain system in which the waste of one field serves as the raw material for the next.
6. Introduction and adoption of new technologies:
By adoption of IFS, famers can link various agro-enterprises like dairy, poultry,
horticulture, floriculture and sericulture etc. which ensures optimum income to adopt latest
technologies to improve their production status.
7. Employment generation:
IFS increase the requirement of labour for performing different activities in different
sectors thus reduces the problem of unemployment. IFS provide enough scope to engage family
labour round the year.
Figure 2: Generalized scheme of integrated Farming system in relation to Sericulture
Constrains in adoption of Integrated Farming System (J. Pushpa, 2010):

1. Lack of well coordinated extension services.
2. Lack of proper demonstration.
3. Lack of composite credit facilities.
74
(ISBN: 978-93-91768-45-4)
4. Lack of awareness among the farmers.

5. Inadequate supply of basic raw material to start any enterprise.
6. Lack of knowledge for proper planting system.
7. Inadequate marketing facilities.
8. Poor health management system in livestock and dairy farming.
9. Lack of irrigation facilities.
10. High mortality rate.
Conclusion:
Integrated Farming in relation to sericulture offers numerous advantages over ordinary
system of mulberry cultivation and silkworm rearing and aims to ensure sustainable agriculture
with profitable outcomes. It extends numerous benefits to the sericulture practising farmers as
they can grasp handsome income via introduction of several part time enterprises including
horticulture, floriculture, fisheries and poultry etc. which enables them to elevate their soco-
economic status as it has tremendous potential for employment generation in rural areas.
Therefore, the concept of integrated farming or mixed farming system should be made familiar
to the sericultural farmers for exponential increase in the crop productivity and elevation in the
farmer‘s income resources with sustainable development of the agriculture.
References:
Baishya, A., Pathak, A. K., Bhowmick, B. C. and Ahmed, S. 2004. Predominent Farming System
and alternative in Assam in Alternative Farming Systems Proceeding of a Symposium held
at PDCSR, Modipuram, Meerut on 16-18 Sept. pp. 228-237.
Chanotra, S., Bali, K. and Bali, R. K. 2019. Sericulture: An opportunity for the uplifment of rural
livelihood. Journal of Entomology and Zoology Studies. 7(6): 1100-1103.
Kerutagi, M. G., Talavar, M. and Pavitra A. S. 2019. Impact of horticulture based integrated
farming system on farmer‘s income and welfare in Northern Karnataka. Journal of
Pharmacognosy and Phytochemistry. 8(3): 1010-1019.
Kumar, S., Dey, A., Kumar, U., Chandra, N. and Bhatt, B. P. 2012. Integrated farming system
for improving agricultural productivity. Status of Agricultural Development in Eastern
India, Publisher: ICAR Research Complex for Eastern Region. pp.205-230
75
Longvah, T., Mangthya, K. and Ramulu, P. 2011. Nutrient composition and protein quality
evaluation of eri silkworm (Samia ricinii) prepupae and pupae. Food Chemistry. 128: 400-
403.
Nagaraju .Y. and Raghavendra. N. 2016. Improve Livelihood Security and Employment
Generation through Integrated Farming System of Scheduled Caste Farm Families in CB
Pura District of Karnataka. International Journal of Science and Research. 5(8): 1419-
1321.
Nigussie, A. 2018. Livestock and Fisheries Research Strategies. Livestock and Fisheries
Research Strategies Poultry, Fisheries, Apiculture and Sericulture (2016 – 2030). Ethiopian
Institute of Agricultural Research.pp.01-227.
Prein M. 2002. Integration of aquaculture into crop–animal systems in Asia. Agricultural
Systems. 71: 127–146.
Pushpa, J. 2010. Constraints in various integrated farming systems. Agriculture Update. 5(3&4):
370-374.
Rai S. 2006. Tasar sericulture, an emerging discipline for conservation and sustainable utilization
of natural resources. The Vision Review Point.
Rames-babu., Ramakrishna, K. S., Harishkumarreddy, Y., Lakshmi, G. and Naidu, N.V. S. 2009.
Metabolic molecular mechanism in silkworm larvae during viral infection: A review.
African Journal of Biotechnology: 8:899- 907.
Rana, S. S. and Rana, M. C. 2011. Cropping System. Department of Agronomy, College of
Agriculture, CSK Himachal Pradesh Krishi Vishvavidyalaya, Palampur, 80 pages.
Yadav, A. K., Nalini, R. and Dashrath, S. 2019. Integrated Farming Systems Approach: Increase
Food Security, Agricultural Farm Income and Rural Economy.
Int.J.Curr.Microbiol.App.Sci. 8(02): 1167-1185.
76
(ISBN: 978-93-91768-45-4)
AN OVERVIEW OF DIFFERENT TYPES OF TOXIC ENVIRONMENTAL

CHEMICALS AND THEIR CELLULAR RECEPTORS
Deeksha Sharma
ICAR- National Dairy Research Institute,
Karnal-132001 (Haryana) India
Abstract:
Chemicals that have ended up in the environment as a result of human activity and are
harmful to human health are known as environmental pollutants. The recent increase in the
presence of these compounds in the environment has raised concerns about human safety.
Humans are exposed to these contaminants on a daily basis through polluted air, soil, and food.
Dioxins, pesticides, plasticizers, and heavy metals are only a few of the many compounds that
make up environmental contaminants. These substances have a negative impact on human
health, including endocrine abnormalities, neurological system problems, infertility, cancer,
immune system disorders, metabolic diseases, and respiratory disorders. The harmful effects of
these chemicals are mediated at the cellular level via their interactions with cellular receptors. In
light of this, the current review aims to consolidate information on the cellular interactions of
environmental chemicals. Previous research has revealed that these compounds have a wide
range of mechanisms of action, which vary depending on the cell model, species, chemical
concentration, and exposure period. The majority of environmental hazardous substances, acted
through nuclear receptors. A vast number of compounds, in particular, acted through the aryl
hydrocarbon receptor. It is generally known that the arylhydrocarbon receptor plays a role in the
toxicity of environmental toxins. Overall, this study focused on the critical receptors that are
known to mediate chemical toxicity. This effort will help us understand the toxic mechanism of
chemicals in a better way.
Keywords: Environmental chemicals, cellular receptors, dioxins, pesticides, heavy metals
Introduction:
Global concern regarding the sources, sinks, distributions, fates, and impacts of
environmental toxins has grown in recent decades. Because of these pollutants' pervasive
presence in the environment and their negative impacts on the environment, and human health,
77
understanding of their toxic mechanism is needed. Briefly, environmental pollutants are

described any physical, chemical, biological, or radiological compound that negatively impact
the air, water, soil quality, or living beings (Steve et al., 2005). In the present review we are
mainly concerned about chemical pollutants. It primarily consists of three chemical classes:
industrial chemicals, agrochemicals, and heavy metals (National Research Council, 2006; Sexton
et al., 2011; CDC). The industrial chemicals contain dioxins, PCBs, and plasticizers whereas
agrochemicals mainly consist of pesticides. The animal and human body are regularly exposing
to these chemicals either directly from foodstuffs or indirectly by the environment. Though all
environmental chemicals are not harmful to human, the body has effective detoxification
mechanism through which it transformed the toxic compounds to less toxic form and then
excrete them (Murphy, 2001). But, some of them are resistant to metabolism, and due to their
stable nature, they persist in the body for a long time, usually called a persistent organic
pollutant. Some of the toxic environmental chemicals are dioxins, polychlorinatedbiphenyls
(PCBs), phthalates, bisphenols (BPA), organochlorine and organophosphate pesticides and heavy
metals such as lead, cadmium and arsenic. These chemicals adversely affect the human health
causing endocrine disruption, immune system disorders, cancer, nervous system damage,
reproductive problems, respiratory disorders and metabolic diseases (Turusov et al., 2002,
Lemaire et al., 2004; Jamal et al., 2013; White and Birnbaum, 2009; Costa, 2008). There are
several available studies described the toxic effects of these chemicals. The present review is
specifically focused on the cellular mechanism of toxicity of environmentally relevant chemicals.
To be more specific, this study will describe the different cellular receptors found to be involved
in the toxicity of environmental chemicals. It could help to understand their toxic effects and
subsequently help in improving their toxicity profiles.
Types of environmental chemicals:

Environmental chemicals are defined as toxic chemicals that are present in a higher
concentration than normal concentration. It may be due to human activity and has a negative
impact on the environment and living beings. In includes different classes of chemicals such as
dioxins, PCBs, pesticides, plasticizers and heavy metals.
1. Dioxins
Dioxins Among them, dioxins are the family of structurally and chemically related
polyhalogenated aromatic hydrocarbons consists of 75 polychlorinated dibenzo-p-dioxin
congeners (PCDD) and 135 polychlorinated dibenzofurans (PCDF) congeners (COMMISSION
DIRECTIVE 2006/13/EC February 2006). Among these 210 chemicals, 17 chemicals are
reported for their human toxic effect. Dioxins have been released into the environment
78
(ISBN: 978-93-91768-45-4)
accidentally in the course of industrial practices such as the production of pesticides and chlorine
bleaching in the paper industry. The term dioxin commonly used for the most toxic
representative of this class is TCDD or 2, 3, 7, 8-tetrachloro-dibenzo P-dioxin. It is a known
cancer-causing agent (NIEHS (National Institute of Environmental Health Sciences) 2001).
2. Polychlorinated biphenyls (PCBs)
Another, toxic group of organic compounds is polychlorinated biphenyls (PCBs). The
PCBs are long-lasting chemical compounds that were widely employed in the 1930s and 1940s.
PCBs have contaminated different environmental matrices worldwide due to their extensive
usage (Borja et al., 2005). It comprises dioxin-like polychlorinated biphenyls (DL-PCBs) and
non-dioxin-like polychlorinated biphenyls (NDL-PCBs). These compounds have long-range
transport and bioaccumulation efficiency (Jing et al., 2018).
3. Pesticides
Pesticides are being used to control the pests in the crops and to improve the quality of
the crops all around the world. The term pesticide comprises herbicide, insecticide, nematicide,
bactericide, insect repellent, and fungicide. Pesticide residues that have been reported in milk
include chlorinated pesticides, organophosphates, herbicides, fungicides, and insecticides
(Aytenfsu et al., 2016). Among pesticides, the most predominant and toxic class is
organochlorine pesticides. Organochlorine pesticides (OCPs) are organic compounds known for
their toxicity, lipophilic properties, and bioaccumulation potential (Harrison, 2007). Most of the
countries of the world prohibited or restricted their usage but still, their microbial degradation
resistant, lipophilic and persistent nature allows them to exist in the environment (Erdoğrul et al.,
2004; Kaushik and Kaushik, 2007).Organochlorine is a class of pesticide widely includes
harmful contaminates, sub-class like dichlorodiphenylethanes, the chlorinated cyclodienes
(aldrin, dieldrin, heptachlor, etc.), and the hexachlorocyclohexanes (lindane). These compounds
had been consumed in the agricultural field to control pests and insects causing tropical diseases
such as malaria, Chagas disease from 1940 to 1990 (Santos et al., 2015). Seeing the presence of
these contaminants in the food products their use in agricultural work was restricted. Despite
that, some of these insecticides are still used as a controlling agent of tropical diseases in some
countries (D Amato et al., 2002; Santos et al., 2015). Persistent organic pollutants list prepared
under Stockholm Convention contain 12 highly toxic chemicals of which 9 are organochlorine
pesticides (UNEP, 2010). Some of the OCPs are known as endocrine disruptors like dieldrin and
heptachlor (Nassar et al., 2016) while some compounds such as dichlorodiphenyltrichloroethane
(DDT), hexachlorocyclohexanes (HCH), are recognized as a carcinogenic chemical to humans
(The International Agency for Research on Cancer (IARC) 2006). Other than them
79
organophosphate are also widely used broad-spectrum pesticides. They have a very specific toxic
mechanism mediated through inhibition of acetyl cholinesterase enzyme (Kazemi et al., 2012).
4. Plasticizers and bisphenol
A plasticizer is a substance that is applied to another substance (typically a plastic or an
elastomer) to make it softer or more malleable. Across 100 distinct plasticizers are produced
around the world today, with phthalate ester plasticizers accounting for around 85 percent of
them (Godwin, 2017). Another synthetic compound used as plasticizer is bisphenol (BPA). It is
the most abundant synthetic compound used worldwide for the production of water and infant
feeding bottles and containers for the storage of food products. It has been established that BPA
can be liberated from containers and migrate into the food products kept in them (Food
Standards Agency. 2001). It is known endocrine disruptor and negatively impact the human
health (Rochester, 2013).
5 Heavy metals
Chemically ―a metal having a density more than 5 g/cm3" is called heavy metal (Csuros
and Csuros, 2002). Trace essential, valuable, radio nuclei, and very poisonous metals are among
them. Copper, iron, zinc, selenium, and cobalt are essential heavy metals that play a vital part in
a variety of biological processes in humans (Mertz, 1981). The phrase "heavy metal," on the
other hand, refers to metals that contaminate food and cause harm even in trace amounts (Duffus,
2002). Such toxic heavy metals include lead, mercury, arsenic, and cadmium thatare highly
dispersed in the environment. These metals are used in a variety of industrial processes such as
metal plating, textiles, battery manufacturing, metallurgy, chemical manufacturing, automobiles,
and petroleum (Arruti, 2010, Sträter et al., 2010).
Route of exposure of environmental chemicals:

Environmental pollutants are widely present in the air, soil, and/or water (Maria and
Mary, 2012). Some of these compounds are very stable and have a long half-life and high
persistency in nature (Jorgenson, 2001). Previous study has shown that agricultural and
horticultural fields‘ soil is heavily contaminated with organochlorines and dioxins (Hilber et al.,
2008.) These persistent chemicals easily passed from the agricultural fields‘ soil to the crops and
subsequently to the animals. After metabolization in animals, most of the chemicals are
transformed into excretory form and removed from the system. Yet some persistent contaminants
are stored in the fatty tissue of the body and move to the circulation and subsequently reappear in
the milk and animal products (Martínez et al., 1997). Humans are directly exposed to these
chemicals through contaminated air, soil, water and indirectly via contaminated animal products,
80
(ISBN: 978-93-91768-45-4)
fruits, vegetables and foodstuffs (Jayaraj et al., 2016; Wang and Needham, 2007;Wilkowska and
Biziuk, 2011).
Mechanism of action at cellular level:

Environmental chemicals have a variety of effects on physiological cell processes,
resulting in changes in normal cell function, particularly at the molecular and biochemical level
(Dizer et al., 2001). Humans get exposed to these chemicals through different routes and within
human these compounds distribute to the whole body mainly through circulation or accumulates
in fat-rich tissues. In addition, most of the pollutants can permeate into the cell by passive
diffusion due to their lipophilic nature and can interact with cell receptors and subsequently alter
the cellular environment.Several studies reported that xenobiotics predominately act through
nuclear receptors such as aryl hydrocarbon receptor (AhR), estrogen receptor (ER), pregnane X
receptor (PXR), and constitutive androstane receptor (CAR). Nuclear receptors (NRs) are present
in the cytosol in the inactive complex form and when lipophilic pollutant/ligand binds with them,
NRs get activated and transferred to the nucleus. In the nucleus, these receptors bind with the
xenobiotic responsive element in the genes and alter the transcription of downstream genes. In
general, NRs regulate the important physiological process of the cell such as growth,
development, and metabolism.
1. Arylhydrocrabon receptor
Among all NRs most prominent receptor responsive to the toxic action of environmental
chemicals is the aryl hydrocarbon receptor (AhR). It controls the transcription of genes involved
in xenobiotic metabolism and cellular proliferation (Lewis et al., 2006). Concerning the affinity
of chemicals to AhR, industrial chemicals were screened virtually using bioinformatics tools
such as ligand and structure-based methods. Structurally similar small halogenated and
polycyclic aromatic hydrocarbon compounds were selected from the AhR modulator database.
The obtained result showed that 41 compounds among 429 selected compounds had an affinity
to the AhR. Further, it was found that among 41 probable ligands, 4 compounds already
confirmed their AhR affinity during in vitro assays in humans (Larsson et al., 2018). Likewise,
the affinity of non -planar PCBs such as PCBs- 99, 138, 163, 180, and 194 were checked in
human liver cells and their stimulatory effect on the PXR and CAR was observed (Salman and
Plant, 2012). The activity of lower chlorinated PCB congeners was also checked against various
nuclear receptors. Interestingly obtained results showed the dose-dependent effect of tested
PCBs, like in nanomolar concentration most of them acts as anti-androgenic compounds while in
micromolar concentration acts as estrogenic compound PCBs 8, 28 and 31 inhibit the AhR while
81
PCB- 4, 8, and 18 were found agonist to CAR and PXR (Pencikova et al., 2018). Similarly,
Takeuchi and coworkers screened the 200 pesticides against AhR using DR-EcoScreen cells.
Among all tested pesticides, 11 pesticides showed the AhR-agonist activity. It includes
acifluorfen-methyl, bifenox, chlorpyrifos, chlorpropham, diurondiethofencarb, linuron,
isoxathion, quinalphos, propanil, and prochloraz. Further, their structural similarity analysis
showed that three herbicides propanil, diuron, and linuron have a strong affinity to AhR due to
their similar chemical structure like AhR natural ligand. This AhR agonistic activity of selected
herbicides was validated in the in vivo model and results showed the stimulatory effect of these
herbicides on AhR markers genes CYP1A1 and CYP1A2 (Takeuchi et al., 2008). In case of
heavy metals their mechanism is not clear. However, apparently it is established that lead is
acting through AhR. Effect of Industrial chemicals like lead and mercury were investigated at
transcriptional level on various nuclear receptors. Amusingly, it was found that these chemicals
increase the mRNA of AhR receptor and ARNT protein. Further, AhR responsive genes like
CYP1A1 and CYP1B1 were also checked and found significantly upregulated in the test subject
(Cavallini et al., 2016). Moreover, two metal responsive elements (MREs) have been identified
in the CYP1A1 promoter region, suggesting the involvement of this gene in lead cellular
mechanism (Williams, 2000). In another study CYP1A1 expression was analyzed in response to
lead and TCDD and finding shows that lead induces CYP1A1 gene expression alone or
synergistically with TCDD, by forming AHR/ARNT/XRE complex (Korashy et al.,
2005). Overall, AhR is the primary receptor that mediates the toxicity of dioxins and some
pesticides.
2. Estrogen receptor
Another important receptor involves in the toxicity of xenobiotics is the estrogen receptor
(ER). It exists in two different isoforms of estrogen receptor alpha (ERα) and estrogen receptor
beta (ERβ). Among both the isoforms, ERα is predominantly involved in the estrogenic activity
of different compounds and works as a ligand-dependent transcription factor (Jordan and
O‘Malley, 2007). ERα affinity is not limited to only natural ligands but it also binds with
estrogenic xenobiotics/endocrine disruptors (Sengupta and Jordan, 2008). This is well supported
by a previous study, in which 200 pesticides and their isomers were tested for estrogenic activity
against human ER isoforms (hERα and hERβ) along with human androgen receptor (hAR) in
Chinese hamster ovary cells. It was found that 47 pesticides had hERα affinity and 33 showed
hERβ affinities while 29 pesticides had agonist activity against both the ER isoforms. In the case
of hAR, none of the tested pesticides showed hAR mediated activity Kojima et al., 2004. In
addition, differential interaction of BSA with ERα and ERβ was checked in Hela cells and found
82
(ISBN: 978-93-91768-45-4)
that BPA has a dose-dependent estrogenic activity to ERα (Hiroi et al., 1999). This is correlated
with BPA‘s endocrine disruptor activity (Krishnan et al., 1993).
Table 1: Environmental chemicals and their receptors
Receptors Xenobiotics Action Reference
ERα Anilofos, chlorpyrifos, bromophos ethyl & Agonist Kojima et
methyl, dichlorofenthion, EPN, ethion, al.,2004
fenitrothion, isofenphos, isoxathion, leptophos, Stoica et al.,
phenothoate, prothiofos, quinalphosaldrin, 2000
dieldrin, alpha, beta & sulfate endosulfan, Morales et al.,
endrin, heptachlor epoxide, methoxychlor, 1994
methiocarb, cyfluthrin, cypermethrin, Choe et al.,
fenvalerate, flucythrinate, chloronitrofen, DDT 2003
and DDE, Cadmium and arsenic
ERβ Leptophos, prothiofos, quinalphos, tolchlofos Agonist Kojima et
methyl, alpha & beta endosulfan, sulfate al.,2004
endosulfan, heptachlor epoxide & heptachlor
Dioxins, HCH, aciflurofen methyl, bifenfox, Agonist Janoseket al.,
AhR chlorprofos,chlorpropham, diethofencarb, 2006
isoxanthion, quinalphos, propanil, diuron, Takeuchi et al.,
linuron, prochloraz, and lead (Pb) 2008
PCB118, PCB 138, PCB 28, HCB, gamma- Antagonist Doan et al.,
HCH, dieldrin and chlordane 2019
Aldrin, dieldrin, alpha, beta &sulfate Agonist Klieweret

PXR endosulfan, endrin, heptachlor epoxide, al.,2002
methoxychlor, PCBs- 4,8 & 18, BPA, Mono-2 Sui et al., 2012
ethylhexyl phthalate Hurst and
Waxman, 2004
AR Parathion, cypermethrin, aldrin, endrin, dieldrin, Antagonist Xu et al., 2008
chlordane, endosulfan and organophosphate Lemaire et al.,
fenitrothion 2004
Tamura et al.,
2003
Dieldrin, PCBs-4,8 18, 99, 138, 153 180 & 194 Agonist Salman et al.,
CAR 2012, Wang et
al 2020
GCR Arsenic Antagonist Kaltreider et
al.,2001 Sargis
BPA, endrin, dicylohexyl phthalate Agonist
et al., 2018
83
3. Other nuclear receptors

Based on these studies, it can be established that xenobiotics predominately act through
the aryl hydrocarbon receptor and estrogen receptor alpha. However, few xenobiotics also bind
with androgen receptor (AR), pregnane X receptor (PXR), and constitutive androstane receptor
(CAR). The androgen receptor (AR) is a key transcriptional regulator involved in male
reproduction and development. The chemicals causing the negative effects to humans might be
due to AR binding inhibition and subsequent AR-dependent transcription (Xu et al., 2008).
Similarly, the nuclear receptor family of ligand-activated transcription factors includes the
pregnane X receptor (PXR), which is an important part of the body's defense against harmful
xenobiotics. A wide range of lipophilic xenobiotics, including as prescription medicines, herbs,
pesticides, endocrine disruptors, and other environmental pollutants, activate PXR (Kliewer,
2003). Another nuclear receptor CARis activated by a wide range of chemicals and it found to be
involved inthe transcription of genes of xenobiotic metabolism pathway (Maglich et al., 2002).
One more nuclear receptor which mediate the toxic effects of theses is glucocorticoid receptor
(GCR). The table given below summarized the different nuclear receptors, their xenobiotic
ligands and their agonist or antagonist action (Table 1).
Conclusion:
Most of the environmental chemicals are act through nuclear receptors at the cellular
level. Among all nuclear receptors, arylhydrocarbon receptor and estrogen receptor alpha
mediate the toxicity of most the dioxins and pesticides respectively. Some chemicals may act
through more than one receptor on the other hand two or more chemicals are act through one
receptor. However, a specific mechanism can‘t be delineated for a chemical as it may vary
depending on cellular model, period and amount of chemical exposure. More research is needed
to impart specific knowledge on the toxic mechanism of chemicals.
References:
Al-Salman, F., and Plant, N. (2012). Non-coplanar polychlorinated biphenyls (PCBs) are direct
agonists for the human pregnane-X receptor and constitutive androstane receptor, and
activate target gene expression in a tissue-specific manner. Toxicology and applied
pharmacology, 263(1), 7-13.
Arruti, A., Fernández-Olmo, I., andIrabien, Á. (2010). Evaluation of the contribution of local
sources to trace metals levels in urban PM2. 5 and PM10 in the Cantabria region (Northern
Spain). Journal of environmental monitoring, 12(7), 1451-1458.
84
(ISBN: 978-93-91768-45-4)
Aytenfsu, S., Mamo, G., and Kebede, B. (2016). Review on chemical residues in milk and their
public health concern in Ethiopia. J Nutr Food Sci, 6(4), 524-524.
Borja, J., Taleon, D. M., Auresenia, J., and Gallardo, S. (2005). Polychlorinated biphenyls and
their biodegradation. Process biochemistry, 40(6), 1999-2013.
Cavallini, A., Lippolis, C., Vacca, M., Nardelli, C., Castegna, A., Arnesano, F., ... andDepalo, R.
(2016). The effects of chronic lifelong activation of the AHR pathway by industrial
chemical pollutants on female human reproduction. PloS one, 11(3), e0152181.
Choe, S. Y., Kim, S. J., Kim, H. G., Lee, J. H., Choi, Y., Lee, H., and Kim, Y. (2003).
Evaluation of estrogenicity of major heavy metals. Science of the total environment, 312(1-
3), 15-21.
Costa, L. G. (2008). Toxic effects of pesticides. Casarett and Doull’s toxicology: the basic
science of poisons, 8, 883-930
Csuros, M. and Csuros, C., 2016. Environmental sampling and analysis for metals. CRC Press.
D'amato, C., Torres, J. P., andMalm, O. (2002). DDT (diclorodifeniltricloroetano): toxicidade e
contaminaçãoambiental-umarevisão. Química Nova, 25(6a), 995-1002.
Dizer, H., de Assis, H. D. S., and Hansen, P. D. (2001). Cholinesterase activity as a bioindicator
for monitoring marine pollution in the Baltic Sea and the Mediterranean Sea.
In Biomarkers in Marine Organisms (pp. 331-342). Elsevier Science.
Doan, T. Q., Berntsen, H. F., Verhaegen, S., Ropstad, E., Connolly, L., Igout, A., ... andScippo,
M. L. (2019). A mixture of persistent organic pollutants relevant for human exposure
inhibits the transactivation activity of the aryl hydrocarbon receptor in
vitro. Environmental Pollution, 254, 113098.
Dos Santos, J. S., Schwanz, T. G., Coelho, A. N., Heck-Marques, M. C., Mexia, M. M.,
Emanuelli, T., andCostabeber, I. (2015). Estimated daily intake of organochlorine
pesticides from dairy products in Brazil. Food Control, 53, 23-28.
Duffus, J.H., 2002. ―Heavy metals" a meaningless term?(IUPAC Technical Report). Pure and
applied chemistry, 74(5), pp.793-807.
Erdoĝrul, Ö., Covaci, A., Kurtul, N., and Schepens, P. (2004). Levels of organohalogenated
persistent pollutants in human milk from Kahramanmaraş region, Turkey. Environment
international, 30(5), 659-666.
Garcia-Morales, P., Saceda, M., Kenney, N., Kim, N., Salomon, D. S., Gottardis, M. M., ... and
Martin, M. B. (1994). Effect of cadmium on estrogen receptor levels and estrogen-induced
85
responses in human breast cancer cells. Journal of Biological Chemistry, 269(24), 16896-
16901.
Godwin, A. D. (2017). Plasticizers. In Applied plastics engineering handbook (pp. 533-553).
William Andrew Publishing.
Harrison, S. (1997, November). Organochlorines in Australia. In Proceedings, UNEP Workshop
on Persistent Organic Pollutants (POPs), Bangkok.
Hilber, I., Mäder, P., Schulin, R., and Wyss, G. S. (2008). Survey of organochlorine pesticides in
horticultural soils and there grown Cucurbitaceae. Chemosphere, 73(6), 954-961.
Hiroi, H., Tsutsumi, O., Momoeda, M., Takai, Y., Osuga, Y., andTaketani, Y. (1999).
Differential interactions of bisphenol A and17β-estradiol with estrogen receptor α (ERα)
and ERβ. Endocrine journal, 46(6), 773-778.
Https://www.cdc.gov/biomonitoring/environmental_chemicals.html
Hurst, C. H., and Waxman, D. J. (2004). Environmental phthalate monoesters activate pregnane
X receptor-mediated transcription. Toxicology and applied pharmacology, 199(3), 266-
274.
IARC, (2018) Working Group on the Evaluation of Carcinogenic Risk to Humans, DDT,
Lindane, and 2,4-D. IARC Monographs. 113:5132018.
Jamal, Q., Durani, P., Khan, K., Munir, S., Hussain, S., Munir, K., andAnees, M. (2013). Heavy
metals accumulation and their toxic effects. Journal of Bio-Molecular Sciences
(JBMS), 1(1), 27-36.
Jayaraj, R., Megha, P., andSreedev, P. (2016). Review article. Organochlorine pesticides, their
toxic effects on living organisms and their fate in the environment. InterdiscipToxicol 9:
90–100.
Jing, R., Fusi, S., andKjellerup, B. V. (2018). Remediation of polychlorinated biphenyls (PCBs)
in contaminated soils and sediment: state of knowledge and perspectives. Frontiers in
Environmental Science, 6, 79.
Jordan, V. C., and O'Malley, B. W. (2007). Selective estrogen-receptor modulators and
antihormonal resistance in breast cancer. Journal of clinical oncology, 25(36), 5815-5824.
Jorgenson, J. L. (2001). Aldrin and dieldrin: a review of research on their production,
environmental deposition and fate, bioaccumulation, toxicology, and epidemiology in the
United States. Environmental health perspectives, 109(suppl 1), 113-139.
Kaltreider, R. C., Davis, A. M., Lariviere, J. P., and Hamilton, J. W. (2001). Arsenic alters the
function of the glucocorticoid receptor as a transcription factor. Environmental health
perspectives, 109(3), 245-251.
86
(ISBN: 978-93-91768-45-4)
Kaushik, P., and Kaushik, G. (2007). An assessment of structure and toxicity correlation in
organochlorine pesticides. Journal of Hazardous Materials, 143(1-2), 102-111.
Kazemi, M., Tahmasbi, A. M., Valizadeh, R., Naserian, A. A., andSoni, A. (2012).
Organophosphate pesticides: A general review. Agricultural science research journals, 2.
Kliewer, S. A. (2003). The nuclear pregnane X receptor regulates xenobiotic detoxification. The
Journal of nutrition, 133(7), 2444S-2447S.
Kojima, H., Katsura, E., Takeuchi, S., Niiyama, K., and Kobayashi, K. (2004). Screening for
estrogen and androgen receptor activities in 200 pesticides by in vitro reporter gene assays
using Chinese hamster ovary cells. Environmental health perspectives, 112(5), 524-531.
Korashy, H. M., and El-Kadi, A. O. (2005). Regulatory mechanisms modulating the expression
of cytochrome P450 1A1 gene by heavy metals. Toxicological Sciences, 88(1), 39-51.
Krishnan, A. V., P. Stathis, S. F. Permuth, L. Tokes, and D. Feldman.(1993). Bisphenol A: an
estrogenic substance is released from polycarbonate asks during autoclaving.
Endocrinology 132:2279–2286
Larsson, M., Fraccalvieri, D., Andersson, C. D., Bonati, L., Linusson, A., and Andersson, P. L.
(2018). Identification of potential aryl hydrocarbon receptor ligands by virtual screening of
industrial chemicals. Environmental Science and Pollution Research, 25(3), 2436-2449.
Lemaire, G., Terouanne, B., Mauvais, P., Michel, S., andRahmani, R. (2004). Effect of
organochlorine pesticides on human androgen receptor activation in vitro. Toxicology and
applied pharmacology, 196(2), 235-246.
Lemaire, G., Terouanne, B., Mauvais, P., Michel, S., andRahmani, R. (2004). Effect of
organochlorine pesticides on human androgen receptor activation in vitro. Toxicology and
applied pharmacology, 196(2), 235-246.
Lewis, N. A., Williams, T. D., and Chipman, J. K. (2006). Functional analysis of a metal
response element in the regulatory region of flounder cytochrome P450 1A and
implications for environmental monitoring of pollutants. Toxicological Sciences, 92(2),
387-393.
Magnusson, U., andLjungvall, K. (2014). Environmental pollutants and dysregulation of male
puberty—A comparison among species. Reproductive Toxicology, 44, 23-32.
María, C. L. and Mary, T. (2012) Chemical Residues in Animal Food Products: An Issue of
Public Health, Public Health Methodology, Environmental and Systems Issues. National
University of Colombia, Colombia,51,641-648
87
Martínez, M., Angulo, R., Pozo, R., andJodral, M. (1997). Organochlorine pesticides in
pasteurized milk and associated health risks. Food and chemical toxicology, 35(6), 621-
624.
Mertz, W., 1981. The essential trace elements. Science, 213(4514), pp.1332-1338.
Murphy, P. J. (2001). Xenobiotic metabolism: a look from the past to the future. Drug
Metabolism and Disposition, 29(6), 779-780.
National Research Council. (2006). Human biomonitoring for environmental chemicals. National
Academies Press.
Pěnčíková, K., Svržková, L., Strapáčová, S., Neča, J., Bartoňková, I., Dvořák, Z., ... and
Machala, M. (2018). In vitro profiling of toxic effects of prominent environmental lower-
chlorinated PCB congeners linked with endocrine disruption and tumor
promotion. Environmental Pollution, 237, 473-486.
Rochester, J. R. (2013). Bisphenol A and human health: a review of the literature. Reproductive
toxicology, 42, 132-155.
Sargis, R. M., Johnson, D. N., Choudhury, R. A., and Brady, M. J. (2010). Environmental
endocrine disruptors promote adipogenesis in the 3t3‐l1 cell line through glucocorticoid
receptor activation. Obesity, 18(7), 1283-1288.
Sengupta, S., and Jordan, V. C. (2008). Selective Estrogen Modulators as an Anticancer
Tool. Innovative Endocrinology of Cancer, 206-219.
Sexton, K., Ryan, A. D., Adgate, J. L., Barr, D. B., and Needham, L. L. (2011). Biomarker
measurements of concurrent exposure to multiple environmental chemicals and chemical
classes in children. Journal of Toxicology and Environmental Health, Part A, 74(14), 927-
942.
Steve J. D‘Surney, Mike D. Smith (2005). Chemicals of Environmental Concer. Encyclopedia
of Toxicology (Second Edition).
Stoica, A., Katzenellenbogen, B. S., and Martin, M. B. (2000). Activation of estrogen receptor-α
by the heavy metal cadmium. Molecular Endocrinology, 14(4), 545-553.
Sträter, E., Westbeld, A., and Klemm, O. (2010). Pollution in coastal fog at Alto Patache,
northern Chile. Environmental Science and Pollution Research, 17(9), 1563-1573.
Sui, Y., Ai, N., Park, S. H., Rios-Pilier, J., Perkins, J. T., Welsh, W. J., and Zhou, C. (2012).
Bisphenol A and its analogues activate human pregnane X receptor. Environmental health
perspectives, 120(3), 399-405.
Takeuchi, S., Iida, M., Yabushita, H., Matsuda, T., and Kojima, H. (2008). In vitro screening for
aryl hydrocarbon receptor agonistic activity in 200 pesticides using a highly sensitive
88
(ISBN: 978-93-91768-45-4)
reporter cell line, DR-EcoScreen cells, and in vivo mouse liver cytochrome P450-1A
induction by propanil, diuron and linuron. Chemosphere, 74(1), 155-165.
Tamura, H., Yoshikawa, H., Gaido, K. W., Ross, S. M., DeLisle, R. K., Welsh, W. J., and
Richard, A. M. (2003). Interaction of organophosphate pesticides and related compounds
with the androgen receptor. Environmental health perspectives, 111(4), 545-552.
Turusov, V., Rakitsky, V., andTomatis, L. (2002). Dichlorodiphenyltrichloroethane (DDT):
ubiquity, persistence, and risks. Environmental health perspectives, 110(2), 125-128.
Wang, R. Y., and Needham, L. L. (2007). Environmental chemicals: from the environment to
food, to breast milk, to the infant. Journal of Toxicology and Environmental Health, Part
B, 10(8), 597-609.
White, S. S., and Birnbaum, L. S. (2009). An overview of the effects of dioxins and dioxin-like
compounds on vertebrates, as documented in human and ecological epidemiology. Journal
of Environmental Science and Health, Part C, 27(4), 197-211.
Wilkowska, A., andBiziuk, M. (2011). Determination of pesticide residues in food matrices
using the QuEChERS methodology. Food chemistry, 125(3), 803-812.
Williams, T. D., Lee, J. S., Sheader, D. L., and Chipman, J. K. (2000). The cytochrome P450 1A
gene (CYP1A) from European flounder (Platichthysflesus), analysis of regulatory regions
and development of a dual luciferase reporter gene system. Marine Environmental
Research, 50(1-5), 1-6.
Xu, L. C., Liu, L., Ren, X. M., Zhang, M. R., Cong, N., Xu, A. Q., and Shao, J. H. (2008).
Evaluation of androgen receptor transcriptional activities of some pesticides in
vitro. Toxicology, 243(1-2), 59-65.
89
HEAT STRESS AND ITS IMPACT ON HORMONAL PHYSIOLOGY OF

HOMEOTHERMIC ANIMALS
Manomita Maity1 and Debajyoti Pradhan*2
1Department of Life Sciences, Presidency University, Kolkata, West Bengal
2Department of Zoology, Egra Sarada Shashi Bhusan College, Purba Medinipur, West Bengal
Abstract:
Stress is either a short-term or long-term unusual condition of the body produced by
physiological, environmental and poor management factors which change the equilibrium
between the body and its environment. An environmental stressor such as heat stress is
detrimental for animal health and animal agriculture also. Health problems like respiratory
infection, poor diet, endocrine disorders, immunosuppression, etc. are the manifestations of
chronic heat stress. Several hormones are involved in the heat stress response like cortisol,
catecholamines, thyroxine, and gonadotropins. Heat stress affects the neuroendocrine profile via
the activation of the Hypothalamo-Pituitary-Adrenal (HPA) axis and the Sympathetic-Adrenal
Medullary (SAM) axis. HPA axis is stimulated in heat-stressed animals to release cortisol,
causing lipolysis and proteolysis for energy production. Adrenaline secretion is elevated in heat-
stressed conditions to increase heart rate and blood pressure to cope up the body with this
situation. Glucocorticoids act on CNS and thereby inhibit TSH secretion, and plasma T3 & T4
levels get decreased to reduce metabolic rate and body heat generation. These hormonal changes
in the body are required for a 'fight-or-flight‘ response. Heat shock proteins (HSPs) such as
Hsp70 and Hsp90 play a predominant role as a biomarker of heat stress. The phytochemicals like
curcumin, lycopene, and antioxidants like Vitamin C are used to treat a heat-stressed victim.
Keywords: Stress, HPA axis, Endocrine disorders, Heat shock proteins, Phytochemicals.
Introduction:
Stress as a general term is used to describe the sum of biological responses or defense
mechanisms of the animal body. According to Selye (1976), ―stress is the nonspecific response
of the body to any demand‖ and a stressor is ―an agent that produces stress at any time‖. Stress is
a situation that tends to disturb the equilibrium between the organism and its environment. Heat
stress results from a negative balance between the net amount of energy flowing from the
90
(ISBN: 978-93-91768-45-4)
animal‘s body to its surroundings and the amount of heat energy generated by the animal. All
animals have a particular zone of thermal comfort that varies from species to species,
physiological status, the velocity of ambient air, or degree of solar radiation (NRC, 1981). When
the environmental temperature exceeds the thermoneutral zone (TN), the animal lives in a warm
zone where thermoregulatory reactions are very limited. But when the environmental
temperature increases beyond the upper critical leval, that results in heat stress in the animal
body (Fig. 1). This imbalance is caused by various factors such as environmental factors like
sunlight, thermal irradiation, air movement, humidity, noise, cold, etc., and physiological factors
like disease, vaccination, changes in feed, nutritional deficiency, etc. (Lara et al., 2013). Another
factor also responsible for stress in livestock animals is a poor management system which
includes improper ventilation, overcrowding of animals, improper handling, and poor
transportation. Further, various agro-climatic zones have variable climates which are also
influenced by seasonal variation throughout the year. Hence, the condition of stress is altered
differently from place to place. Various environmental stressors such as heat stress are
detrimental to animal production and growth rate acceleration in livestock farming (Ghosh et al.,
2017).
Figure 1: Effects of environmental temperature on body heat regulation

(LCT: Lower Critical Temperature, UCT: Upper Critical Temperature)
Several studies have reported that heat stress affects different physiological systems in
animals. Birds are mostly affected by heat stress because they have no sweat glands (Renaudeau
et al., 2012). When the temperature exceeds the comfort level of birds, they lose the ability to
91
efficiently dissipate heat to maintain their homeostasis. This leads to physiological changes that
are related to alterations in hormonal status (Teetar et al., 1985). These physiological alterations
can be summarized as an increase in body temperature, depressed immunity, impairment of
endocrine function, mitochondrial dysfunction, generation of oxidative stress, increase in
transcriptional activity of oxidative genes (Nrf-2 & NF-ҡβ), etc. Animals experiencing heat
stress tend to reduce their stress by producing a special type of protein called Heat Shock Protein
(HSP), which generates some antioxidant molecules at the cellular level (Akbarian et al.,2016)
and activates Nrf-2 downstream signaling pathway (Lee et al.,2019). Many studies reported that
using of phytochemicals are the potent regulator of transcription factors for generating
antioxidant molecule as a stress response (Sahin et al., 2013; Lee et al., 2017).
Impacts of heat stress in the endocrine system:
Responses of stressor play a critical role in the maintenance of homeostasis in animals for
their survival in that environment. Various stressors stimulate the endocrine system, and a wide
variety of homeostatic conditions of the body are maintained mostly through the Hypothalamo-
Pituitary–Adrenal (HPA) axis (Garriga et al., 2006; Star et al., 2008). Stimulation of the HPA
axis and Sympathetic adrenal medullary (SAM) axis through the neuronal circuit enhance the
secretion of Glucocorticoids and Catecholamines as long-term and short-term stress responses
respectively (Fig 2).
Figure 2: Schematic representation of hormonal regulation of stress response
92
(ISBN: 978-93-91768-45-4)
This neuroendocrine system plays a major role as a stress response of the body including
changes in the digestion of food, metabolism rate, heart rate, respiration rate, blood pressure, and
immune responses. Glucocorticoids play a critical role in the regulation of the immune system by
cytokine production mediated through the HPA axis (Webster et al., 2004). Glucocorticoids
permit, stimulate or suppress ongoing stress response and organize the brain for a concomitant
stressor (Sapolsky et al., 2000). Endocrinological changes caused by chronic heat stress in
animals like in broiler, stimulate lipid accumulation through increased de novo lipogenesis,
reduced lipolysis, and enhanced amino acid catabolism (Geraert et al., 1996). Heat stress causes
various hormonal changes in homeothermic animals.
Cortisol:
The actions of cortisol are firmly regulated to ascertain that the body can respond rapidly
to stressful effects and bring back to a normal state just as quickly. In response to various
stressors, corticotropin-releasing factor (CRF) is released from the parvocellular neuron of
paraventricular nuclei (PVN) of the hypothalamus into the hypophyseal portal vein to stimulate
the corticotroph cells of the anterior pituitary gland which release adrenocorticotropic hormone
(ACTH) in the systemic circulation. ACTH binds to the type-2 melanocortin receptor of the
adrenal cortex, thereby releasing glucocorticoids mainly cortisol in the circulation (Chida et al.,
2007). Heat stress-induced animals show an elevated level of glucocorticoids which create
different physiological variations such as increased blood glucose level, enhanced proteolysis,
and glycolysis for providing more energy as a stress response.
A high amount of glucocorticoids reduce immune response as an initial inhibition of
macrophage and T-cell activation. It also suppresses the cellular immunity provided by the
thymus gland but lowers glucocorticoids level that may induce antibody production. This
biphasic role may depend on the blood corticoids concentration (Hall and Goldstein, 1984).
Mineralocorticoids:
Chronic heat stress increases the secretion of mineralocorticoids via the HPA axis. It
causes the retention of water and mineral balance in the body and helps to maintain blood
volume and blood pressure (El-Nouty et al., 1980).
Catecholamines:
As a short-term stress response, heat stress activates the sympathetic innervations of the
adrenal medulla that constitute another neuroendocrine system. Axons, derived from the central
sympathetic neuron innervate chromaffin cells of the adrenal medulla and the synapse release
acetylcholine which evokes the secretion of catecholamines (epinephrine & nor-epinephrine) in
93
the circulation. The molecular mechanism of action of epinephrine is triggered by the G-protein
coupled β-adrenergic receptor which ultimately activates the cAMP pathway for performing
various functions like increasing heart rate, vasodilation of skeletal muscle and blood vessel, and
breaking down of glycogen to glucose for providing more energy to the cell. These physiological
changes are required for the body to adapt to unfavorable situations. This is commonly known as
―fight-or-flight‖ action. This response is also known as the first stage of general adaptation
syndrome (GAS). When the animal's body cannot maintain homeostasis, they undergo a stress
condition. Then GAS helps to return the animal to its normal functioning followed by three
consecutive stages namely alarm, resistance, and exhaustion.
Thyroid Hormones:
Normally body temperature and metabolic activity are regulated by thyroid hormones-
triiodothyronine (T3) and tetraiodothyroxine (T4). Many studies revealed that in heat-stressed
animals, T3 concentration gradually decreases (Elnagar et al., 2010 and Mack et al., 2013) and T4
concentration inconsistently decreases, increases, or has no alteration (Bobek et al.,1980) via
Hypothalamo-pituitary-thyroid axis (HPT). TSH secretion is also inhibited by heat stress to
reduce heat generation and BMR.
Gonadotropins:
Heat stress can also alter the Hypothalamo-Pituitary-Gonadal (HPG) axis and directly
affects reproductive functions. The reduced eggshell quality and egg-laying capacity are
commonly found in female poultry birds that occur by alteration of the normal status of
reproductive hormones and disruption of ovarian function (Rozenboim et al., 2007). In males,
semen volume and its quality get reduced in the heat-stressed animal. Spermatogenesis is sharply
hampered in excessive heat-stressed birds and other animals by disruption of testicular functions
(Joshi et al., 1980). Thermal stress can reduce the concentration of serum LH and lower the
granulosa cell activity in laying hen (Novero et al., 1991). The male rats exposed to chronic heat
stress at the onset of puberty lead to a significant increase in body weight, tail of epididymis, and
prostate gland weights. The exposure of chronic heat stress at post-puberty shows a significant
decrease in body weight, testis, head and tail of epididymis, prostate and seminal vesicle weights
as well as a significant decrease in the level of luteinizing hormone (Ashwaq Hassan, 2019).
High-temperature exposure is detrimental to women's reproductive health. High-temperature
stress affects the menstrual function of career women, such as soldiers, textile workers, and
steelworkers, resulting in a significant increase in the incidence of abnormal menstruation
(GaiHong et al., 2020).
94
(ISBN: 978-93-91768-45-4)
Leptin:
Leptin secretion is relevant in metabolism for energy homeostasis. During heat-stressed
conditions in animals, leptin affects energy metabolism following reducing feed intake by
upregulating fatty acid oxidation and downregulating lipogenesis (Allihaud, 2006 and Rabe et
al., 2008).
Insulin:
Heat stress alters metabolic parameters and may change glucose metabolism and insulin
signaling (Ganesan et al., 2018). Prolonged heat stress (>24 hrs) increases basal and circulating
insulin, and decreases adipose tissue mobilization in a variety of species (Sanz Fernandez et al.,
2015). Insulin is a major anabolic hormone that controls energy functions including glucose and
lipid metabolism. Insulin stimulates glucose uptake through an intracellular signaling cascade
that leads to membrane translocation of Glut4 in skeletal muscle. Heat stress impairs the
sarcolemma Glut4 transporter protein abundance which is consistent with reduced circulating
insulin and feed intake (Ganesan et al., 2018).
Heat stress and cellular homeostasis:
Heat stress is a potent inducer of cellular dysfunction. Most of the cellular energy is
produced by oxidative phosphorylation in mitochondria but chronic heat stress leads to
mitochondrial dysfunction, downsizing of mitochondrial metabolic oxidative pathway, increase
in production of reactive oxygen species (ROS), and alteration in the pattern of antioxidant
enzyme activities which ultimately affect the downstream cell signaling pathway of Nrf-2 and
NF-Ҡβ (Akbarian et al., 2016). Heat stress also causes lipid peroxidation, cytoskeleton assembly
alteration, and protein unfolding in the endoplasmic reticulum.
Cells from all organisms respond to a variety of stresses by rapid synthesis of the
polypeptide, known as Heat shock proteins (HSPs). HSPs play important role in normal
conditions as well as in cellular stress such as Hsp70, Hsp90, etc. Many physiological factors
like hyperthermia, oxidative stress, ROS, etc. modulate Hsp70 expression in the cell. Any
stressor is thought to activate the heat shock factor (HSF) which remains bound to HSP in a
normal condition in an inactive form. These HSFs get separated from HSPs in the cytosol and
get activated during phosphorylation by protein kinase and form trimers. These phosphorylated
HSF trimer complexes enter the nucleus and bind to the heat shock element (HSE) of the
promoter region of Hsp70 gene. A new HSP is synthesized in the cytosol after mRNA
transcription from the nucleus. This molecular mechanism revealed that Hsp70 plays a critical
95
role in the development of thermotolerance and protection from stress-induced cellular damage
(Kevin. C. Kregel, 2002). Hence, HSPs are considered as a potent biomarker of cellular injury.
Role of phytochemicals in heat stress:
Phytochemicals are the products that are derived from plants as secondary metabolites in
the form of dried plant material, extract, or essential oil. Curcumin is a potent bioactive
phytochemical found in turmeric that regulates oxidative stress and anti-inflammatory activity by
its antioxidant capacity (Sahin et al., 2012). Curcumin can alleviate the production of SOD and
GST to prevent oxidative damage of cells in a heat-stressed animal by Nrf-2 gene activation. It
also triggers the production of one of the members of the vitagene family i.e. HSP which is a
potent stress-reducing factor (Sahin et al., 2012). Vitamin C is an antioxidant molecule that
reacts with reactive oxygen, singlet oxygen, and H2O2 with ascorbate peroxidase to neutralize
their toxic effects (Forni et al., 2018). Lycopene is the intermediate substance of carotenoids and
is commonly found in tomatoes. It acts as a scavenger of ROS, lipid peroxyl molecule, nitric
oxide, etc. to prevent cellular damage and inflammatory actions by modulating the gene
transcription of NF-Ҡβ and Nrf-2 activation (Petyaev, 2016 and Sahin et al., 2011). Root ginger
paste is also an effective dietary supplementation applied for reducing heat stress by its
antioxidant and anti-inflammatory properties (Habibi et al., 2014).
Conclusion:
It can be concluded that the hormonal changes help to cope up the animals in heat stress
conditions. These changes are required for the body as a ―fight-or-flight‖ response. These
hormonal changes maintain the body's homeostatic condition by changing physiological
responses such as heart rate, respiration rate, metabolism, etc. This is commonly termed as
General Adaptation Syndrome (GAS). Besides, heat stress can directly hamper the cellular
mechanism by generating ROS, causing oxidative stress as well as protein unfolding, and
modulating different gene transcription activities like Nrf-2 and NF- Ҡβ. Chronic heat stress
generates HSPs which act as a biomarker in heat stress to protect cellular toxicity and reduce
inflammatory actions. One of the most effective techniques to recover the heat-stressed condition
in homeothermic animals is the use of phytochemicals that are present in turmeric powder,
tomato powder, lime juice, root ginger paste, etc. Due to their modulating role on anti-
inflammatory genes and the presence of antioxidant enzymes, phytochemicals protect from
oxidative stress.
96
(ISBN: 978-93-91768-45-4)
Acknowledgment:
The authors are indebted and thankful to Dr. B. R. Maiti, Former Professor, Department
of Zoology, University of Calcutta for his continuous support and guidance to prepare this
article.
References:
Ailhaud G. 2006. Adipose tissue as a secretory organ: from adipogenesis to the metabolic
syndrome. Comptes Rendus Biologies. 329: 570–577.
Akbarian A, Michiels J, Degroote J, Majdeddin M, Golian A. Smet SD. 2016. Association
between heat stress and oxidative stress in poultry; mitochondrial dysfunction and dietary
interventions with phytochemicals. Journal of Animal Science and Biotechnology. 7:37.
Doi 10.1186/s40104-016-0097-5.
Ashwaq Hassan. 2019. Effect of heat stress on sex hormones, sex organ weight and relationships
with sexual puberty in male rats. Iraqi Journal of Veterinary Sciences. 33(2): 63-71.
Chida D. 2007. ―Melanocortin 2 receptor is required for adrenal gland development,
steroidogenesis, and neonatal gluconeogénesis‖.Proceedings of the National Academy of
Sciences of the United States of America. 104.46: 18205-18210.
El-Nouty FD, IM Elbanna, TP Davis, HD Johnson. 1980. Aldosterone and ADH response to heat
and dehydration in cattle. J. Appl. Physiol. Respir. Environ. Exercise Physiol. 48: 249.
Forni C, Facchiano F, Bartoli M, Pieretti S, Facchiano A, D‘Arcangelo D , Norelli S, Valle G,
Nisini R, Beninati S, Tabolacci C, Jadeja R N. 2019. Beneficial Role of Phytochemicals on
Oxidative Stress and Age-Related Diseases. Hindawi BioMed Research Internation. 2019.
https://doi.org/10.1155/2019/8748253.
GaiHong An, Chen X, Li C, Zhang L, Wei M, Chen J, Ma Q, Yang D, Wang J. 2020.
Pathophysiological Changes in Female Rats with Estrous Cycle Disorder Induced by Long-
Term Heat Stress. Biomed res int. vol. 2020. https://doi.org/10.1155/2020/4701563
Ganesan S, Summers CM, Pearce SC, Gabler NK, Valentine RJ, Baumgard LH, Rhoads RP,
Joshua TS. 2018. Short-term heat stress altered metabolism and insulin signaling in
skeletal muscle. J. Anim. Sci. 96:154–167 doi: 10.1093/jas/skx083
Garriga C, Hunter RR, Amat C, Planas JM, Mitchell MA, Moreto M. 2006. Heat stress increases
apical glucose transport in the chicken jejunum. Am. J. Physiol. Reg. Integ. Comp.Physiol.
290: R195–R201.
97
Geraert PA, Padilha JC, Guillaumin S. 1996. Metabolic and endocrine changes induced by
chronic heat exposure in broiler chickens: biological and endocrinological variables. Br. J.
Nutr. 75:205–216.
Ghosh CP, Kesh SS, Tudu NK and Dutta S. 2017. Heat stress in dairy animals- Its Impact and
Remedies:A Review, Int .J. pure app. Biosci. 5(1): 953-965. Doi:
http://dx.doi.org/10.18782/2320-7051.2577
Habibi R, Sadeghi G, Karimi A. 2014. Effect of different concentrations of ginger root powder
and its essential oil on growth performance, serum metabolites and antioxidant status in
broiler chicks under heat stress. Brit Poult Sci. 55:228–37.
Joshi PC, Panda B, Joshi BC. 1980. Effect of ambient temperature on semen characteristics of
White Leghorn male chickens. Indian Vet. J. 57:52–56.
Lara LJ, Rostagno MH. 2013. Impact of heat stress on poultry production. Animals. 3:356–69.
Doi:10.3390/ani3020356
Lee MT, Lin WC, Yu B, Lee TT. 2017. Antioxidant capacity of phytochemicals and their
potential effects on oxidative status in animals - A review. Asian-Australas J Anim Sci.
30:299-308. Doi.org/10.5713/ajas.18.0538.
Lee MT, Lin WC, and Lee TT. 2019. Potential crosstalk of oxidative stress and immune
response in poultry through phytochemicals - A review.Asian-Australas J Anim Sci. 32(
3):309-319.
Nardone A, Ronchi B, Lacetera N, Ranieri MS, Bernabucci U. 2010.Effects of climate changes
on animal production and sustainability of livestock systems. Livestock Sci. 130:57–69.
Nienaber JA, Hahn GL. 2007. Livestock production system management responses to thermal
challenges. Int. J. Biometereol. 52:149–157.
Novero RP, Beck MM, Gleaves EW, Johnson AL, Deshazer JA. 1991. Plasma progesterone,
luteinizing hormone concentrations, and granulose cell responsiveness in heat-stressed
hens.Poult. Sci. 70:2335–2339.
NRC. 1981. Poultry. In: Effect of Environment on Nutrient Requirements of Domestic Animals.
pp. 109–134. National Academy Press, Washington D C.
Petyaev IM. 2016. Lycopene deficiency in ageing and cardiovascular disease. Oxidative
Medicine and Cellular Longevity. 2016(3): 1-6
Rabe K, Lehrke M, Parhofer KG, Broedl UC. 2008. Adipokines and insulin resistance.
Molecular Medicine. 14:741–751.
Renaudeau D, Collin A, Yahav S, De Basilio V, Gourdine JL, Collier RJ. 2012. Adaptation to
hot climate and strategies to alleviate heat stress in livestock production. Anim. 6:707–28.
98
(ISBN: 978-93-91768-45-4)
Rozenboim I, Tako E, Gal-Garber O, Proudman JA, Uni Z. 2007. The effect of heat stress on
ovarian function of laying hens. Poult. Sci. 86:1760–1765.
Sahin K, Orhan C, Tuzcu M, Borawska MH, Jablonski J, Guler O, Sahin N, Hayirli A. 2013.
Berberis vulgaris root extract alleviates the adverse effects of heat stress via modulating
hepatic nuclear transcription factors in quails. Brit J Nutr. 111:609–16.
Sahin K, Orhan C, Smith MO, Sahin N. 2013. Molecular targets of dietary phytochemicals for
the alleviation of heat stress in poultry. World‘s Poult Sci J. 69:113–23.
Sahin K, Orhan C, Akdemir F, Tuzcu M, Ali S, Sahin N. 2011. Tomato powder supplementation
activates Nrf-2 via ERK/Akt signaling pathway and attenuates heat stress-related responses
in quails. Anim Feed Sci Technol. 165:230-7.
Sahin K, Orhan C, Tuzcu Z, Tuzcu M, Sahin N. 2012. Curcumin ameliorates heat stress via
inhibition of oxidative stress and modulation of Nrf2/HO-1 pathway in quail. Food Chem
Toxicol. 50:4035-41.
Sanz Fernandez MV, Stoakes SK, Abuajamieh M, Seibert JT, Johnson JS, Horst EA, Rhoads RP,
Baumgard LH. 2015. Heat stress increases insulin sensitivity in pigs. Physiol. Rep.
3(8):e12478. doi:10.14814/ phy2.12478
Sapolsky RM, Romero LM, Munck AU. 2000. How do glucocorticoids influence stress
responses? Integrating permissive, suppressive, stimulatory, and preparative actions.
Endocr. Rev. 21: 55–89.
Selye H. 1976. Forty years of stress research: principal remaining problems and
misconceptions.Can. Med. Assoc. J. 115: 53–56.
Star L, Decuypere E, Parmentier H.K, Kemp B. 2008. Effect of single or combined climatic and
hygienic stress in four layer lines: 2. Endocrine and oxidative stress responses. Poult. Sci.
87:1031–1038.
Webster JI, Sternberg EM. 2004. ―Role of the hypothalamic pituitary adrenal axis,
glucocorticoids and glucocorticoid receptors in toxic sequelae of exposure to bacterial and
viral products‖. Journal of Endocrinology. 181.2: 207-221.
99
DETERMINATION OF MICROBIAL GUT DENSITY IN FRESH WATER FISHES

Annapurneshwari H
I. D. S. G. Govt. College, Chikkamagaluru, Karnataka
Abstract:
Probiotic bacteria are essential for beneficial effect on particular organism‘s health and
host nutrition for healthy gastrointestinal function. In the present study four economically
important food fishes were collected from fish market of Chikkmagaluru town and microbial gut
density of Oreochromis niloticus, Pangasius bocourti, Catla catla, Ictalurus furcatus were
determined. The microbial gut density of fishes varies from place to place as it depends on the
feeding habit and other physiological attributes of fish. Total viable count (TVC) obtained in
terms of colony forming unit (cfu) from the gut of Oreochromis niloticus is 112×103 which is
~1.12×105 CFU/ml, Pangasius bocourti is 105×103 which is~1.05 105 CFU/ml, Catla catla is
53×104 which is ~5.3×105 CFU/ml, Ictalurus furcatus is 118×104 which is ~1.18×105 CFU/ml
respectively and Gram‘s staining was carried out for the obtained colonies. Comparatively the
microbiome of catla catla showed higher colony count (5.3×105 CFU/ml). The study has
established the basis for future large-scale investigations of the gut microbiota of fishes. In order
to make assumptions about the ecological consequences of microbiome composition, a much
deep and large-scale investigation of the gut microbiota of fish in this region is also necessary.
Keywords: microbiota, Catla catla, Gram staining, fresh water fish, colony count
Introduction:
Fish has been one of the main foods for humans for many centuries and still constitute an
important part of the diet in many countries (Leisner et al., 1995). The short supplies of animal
protein together with the increasing human population have raised the cost of animal protein to a
level almost beyond the reach of the low income group (Ezeri et al., 2001). As a result, there is a
considerable increase in the demand for fish being the cheapest source of animal protein (Ladipo
et al., 1981). These important attributes makes the commodity readily susceptible to microbial
attack particularly bacteria (Adams et al., 1999). Probiotic bacteria are essential for beneficial
effect on particular organism‘s health and host nutrition for healthy gastrointestinal function. The
100
(ISBN: 978-93-91768-45-4)
action of intestinal flora results in vital benefits, including protection against pathogens and
development of Immune system. Fish receive bacteria in the digestive tract from the aquatic
environment through water and food that are populated with bacteria. Being rich in nutrient, the
environment of digestive tract of fish confers a favourable culture environment for the
microorganisms (2). Animals harbour a complex microbial community, consisting of bacteria,
yeast, viruses, archaeans and protozoans, in their gastrointestinal (GI) tract. These microbes
influence various host functions including development, digestion, nutrition, disease resistance
and immunity. One important aim of GI microbiota studies therefore is to give a scientific basis
for developing effective strategies for manipulating GI microbial communities to promote the
host health and improve productivity. The diversity generally increases as the diet of the fish
changes from carnivorous to omnivorous to herbivorous.
The composition also differs due to different environmental conditions.
Acinetobacter, Aeromonas, Flavobacterium, Lactococcus, and Pseudomonas.
Obligate anaerobes Bacteroides, Clostridium, and Fusobacterium, and members of
family Enterobacteriaceae dominate the gut of freshwater species. The guts of marine fish are
dominatedby
Aeromonas, Alcaligenes, Alteromonas, Carnobacterium, Flavobacterium, Micrococcus,
Moraxella, Pseudomonas and Vibrio.
The world‘s water bodies are teeming with microorganisms. It is estimated that 3.6×1030
microbial cells account for more than 90% of the total fresh water biomass. Fish harbors heavy
loads of bacteria on their gut, skin and adhering slime. The bacterial flora of fish show
considerable variation with the environmental conditions in which the fish live. Normally the pH
of fish gut ranges from 7-8. The bacterial commodity in fish gut is very dense compared to
surrounding water, which suggests that gastrointestinal tract (GIT) provides a favourable
ecological niche for survival of microbes. Most of the bacterial species in the gut of fish are non-
culturable and thus several isolation techniques introduced to explore the microbial communities
in gut of fish. The feeding habit of fish strongly influences the gut microbiota. In fish, the
structure, composition and ecological function of gut microbiota are greatly influenced by the
factors from host such as genetics, weight, gender, immunity, This study reviews the current
knowledge on the microbiota in some selected fishes, emphasizing the compilation of results
reported regarding the most frequently observed bacterial genera. Also the influence of
environmental and host factors on the establishment of the bacterial populations that become a
part of gut of the fish and the importance of these communities on host health, development and
101
nutrition. An attempt is made in this study to investigate the bacterial micro flora associated with
selected fresh water fishes of Chikkmagaluru town.

Sample Collection
Four economically important food fishes were collected from fish market of
Chikkmagaluru town, Karnataka, India. The samples were collected freshly and brought to the
laboratory for further bacteriological analysis. The following fish species were selected for the
study due to their availability in the local fish market.
1) Oreochromis niloticus
2) Pangasius bocourti
3) Catla catla
4) Ictalurus furcatus
Isolation of gut microbes

Collected fishes were washed with distilled water to remove the unwanted particles.
Then the fishes were dissected to remove the digestive tract. The digestive tracts were
homogenized by centrifugation. Homogenization is carried out in order to obtain uniform
distribution of cells within the solution. After centrifugation the supernatant was taken and
serially diluted in distilled water in the test tubes to 10-1, 10-2, 10-3 and 10-4. The dilution is pour
plated on nutrient agar ( Nutrient agar media, 5g of Nacl, 5g of peptone, 1g of yeast extract/beef
extract, 15g of agar per 1000ml of distilled water) and incubated for 24hour at 37 0C.After
incubation the colonies that developed on the petri plates were counted. Those counts within 30-
300 colony forming units (cfu) were reported as total viable count (TVC).
Comparing the microbial density among the selected fishes
From the colonies obtained, the total viable count of the bacterial isolates from the gut of
selected fish such as Oreochromis niloticus, Pangasius bocourti, Catla catla and Ictalurus
furcatus is calculated by using the following formula.
Microbial load, CFU/ml =Number of colonies × reciprocal of the dilution factor.
From this formula the total viable count of the gut microbial load in terms of cfu can be
calculated. After having the total count for calculation the obtained colonies were subjected to
Gram staining procedure for the identification of the presence of gram positive and gram
negative bacteria.
102
(ISBN: 978-93-91768-45-4)
Result and Discussion:

In the present study microbial load in the gut region of the fresh water fishes like
Oreochromis niloticus, Pangasius bocourti, Catla catla, Ictalurus furcatus were determined.
The Nile tilapia (locally called as jalebi fish), is a teleost fresh water fish of class
Actinopterygii, family Cichlidae, widely known for its importance in aquaculture. These are
omnivorous, they feed on phytoplankton, and macrophytes, insects, detritus and zooplankton
were the most important food items.
After incubating the petri plates for 24 hours at 37oc, the colonies obtained were taken for
further enumeration of the bacterial density by using the formula to find the total viable count of
bacterial in terms of colony forming unit.
The gut bacterial count obtained from the Oreochromis niloticus fish by serial dilution
are as follows,
330 number of colonies from 10-2 dilution.
According to the above mentioned formula the colony count should range from 30 to 300
so the count within this form the colony forming unit. Therefore 122 from 10 -3 dilution is taken
for consideration, which is; ~ 1.22×105 CFU/ml.
Pangasius bocourti
Pangasius bocourti which belong to class Actinopterygii and family pangasiidae is
locally known as Basa fish. They are fresh water residents, benthopelagic, and potadromus. They
are the bottom feeders. They feed on oligochaete, shrimps, crabs, molluscs, fishes and plants.
The gut bacterial count obtained from Pangasius bocourti fish by serial dilution are as
follows,
313 number of colonies from 10-2 dilutions.
According to the above mentioned formula the colony count should range from 30 to
300. So the colony counts within this form the colony forming unit. Therefore in the obtained
colony counts, the count obtained from the 10-3 dilution will be considered as its ranges above 30
103
and below 300 colonies. From this 105 numbers of colonies from 10-3 dilution make a colony
forming unit and is taken for consideration, which is applied to the formula to get the final result,
that is; ~ 1.05×105 CFU/ml.
Catla catla
Catla catla also known as the major south Asian carp, is an economically important south
Asian freshwater fish in the carp family Cyprinidae. Catla is a surface and mid water feeder.
Adults feed on zooplankton using large gill rakers, but young ones on both zooplankton and
phytoplankton. Catla is comparatively large when compared to other fresh water fishes and they
are Voracious feeders feed on many phytoplankton as well as some zooplanktons. Due to this
property of catla there are the chances of presence of microbial flora in its gut in large quantity.
After incubating the petri plates for 24 hours at 37oc, the colonies obtained were taken for further
enumeration of the bacterial density by using the formula to find the total viable count of
The gut bacterial count obtained from Catla catla fish by serial dilution are as follows,
53 number of colonies from 10-4 dilution
so the colony count within this form the colony forming unit. Therefore in the obtainedcolony
counts, the count obtained from the 10-4 dilution will be considered as its ranges above 30
colonies and below 300 colonies. From this 53 numbers of colonies from 10-4 dilution make a
colony forming unit and is taken for consideration, which is applied to the formula to get the
final result, that is given as; ~ 5.3×105 CFU/ml.
Ictalurus furcatus
Ictalurus furcatus commonly called as blue cat fish is the largest species of North
America. catfish and is spread worldwide reaching length 165cm.Blue cat fish are opportunistic
predators and eat any species of fish they can catch, along with freshwater mussels, frogs and
other readily available aquatic food sources.
The gut bacterial count obtained from Ictalurus furcatus fish by serial dilution are as
follows,
441 number of colonies from 10-2dilution.
104
(ISBN: 978-93-91768-45-4)
346 number of colonies from 10-3dilution.

so the colony count within this form the colony forming unit. Therefore in the obtained colony
counts, the count obtained from the 10-4 dilution will be considered as its ranges above 30
colonies and below 300 colonies. From this 118 numbers of colonies from 10-4 dilution make a
colony forming unit and is taken for consideration, which is applied to the formula to get the
final result, that is given as; ~ 1.18×105 CFU/ml.
Comparing the microbial density among the selected fishes.
The microbial gut density of fishes varies from place to place as it depends on the feeding
habit and other physiological attributes of fish. From the colonies obtained, total viable count
(TVC) obtained in terms of colony forming unit (cfu) from the gut of Oreochromis niloticus is
122×103 which is ~1.22×105 CFU/ml, Pangasius bocourti is 105×103 which is~1.05 105
CFU/ml, Catla catla is 53×104 which is ~5.3×105 CFU/ml, Ictalurusfurcatus is 118×104 which
is ~1.18×105 CFU/ml (Table.2)
Table 1: Comparison of gut microbial density of selected fishes
Colony Count In Each Dilution Total Viable
Fish Species -2 -3 -4
10 10 10 Count (Cfu)
Oreochromis niloticus 330 122 14 1.22× 105
Pangasius bocourti 313 105 28 1.05× 105
Catla catla 221 146 53 5.3 ×105
Ictalurus furcatus 441 346 118 1.18× 105
The bacterial density obtained from all the selected fishes were recorded in the Table 1.
By plotting the graph of selected fishes verses its gut microbial density in terms of cfu, variation
in bacterial total viable count of selected fishes namely Oreochromis niloticus, Pangasius
bocourti, Catla catla and Ictalurus furcatus can be concluded and is shown in fig.1. By plotting
the graph for the obtained data, the comparative analysis of gut microbial density of selected
fishes is made. Comparatively the microbiome of catla catla showed higher colony count
(5.3×105 CFU/ml).
105
GUT MICROBIAL DENSITY OF FISH IN TERMS OF CFU (*10-5)

6
5.3
5
3
GUT MICROBIAL DENSITY
2
1.23 1.18
1.05
1
0
Orochromis niloticusPangasius bocourti Catla catla Ictarulus furcatus
Figure 1: Graph plotted by comparing the gut microbial density of selected fishes
X-axis depicts the fish species, Y-axis depicts the gut microbial density in terms of cfu.
Differentiation of gram positive and gram negative bacteria present in the obtained
colonies. The colonies were observed with different morphology and those which showed
varying outcomes in gram‘s staining technique. The below table depicts the result of gram‘s
staining obtained for the selected fish species.(Table.2)
Table 2: Gram’s staining carried out for the obtained colonies
Possibility of Bacteria
Fish Species
Gram Positive Gram Negative
Oreochromis niloticus Present Present
Pangasius bocourti Present Absent
Catla catla Absent Present
Ictalurus furcatus Present Absent
The colonies which appear large and purple in color (Fig 2), when subjected to grams
staining showed positive to the staining which illustrates the presence of gram positive bacteria.
Some colonies appear translucent with discrete colonies, when subjected to gram staining
showed pinkish colonies (Fig 3) which depicts the presence of gram negative bacteria.
By subjecting the obtained colonies to gram‘s staining technique we majorly interpret the
difference between gram positive and gram negative type of bacteria. In Oreochromis niloticus
we observed both gram positive and gram negative bacteria; in case of Pangasius bocourti we
observed gram positive bacteria and Catla catla showed the presence of gram negative bacteria
106
(ISBN: 978-93-91768-45-4)
and Ictalurus furcatus showed the presence of gram positive bacteria in its gut. The gram
positive and negative bacteria have both developmental and declinable effects on the host
metabolism. The negative effect involves the occurrence of necrosis of tissues of the host body
not only so much it also obstruct the vessels, capillaries of the host thus inhibit the supply of
sufficient blood to the host body and in turn deplete the oxygen supply to the body, thus the host
will not be able to have a proper developmental aids and it leads to the death of the individual.
Figure 2: Gram positive bacteria Figure 3: Gram negative bacteria
Conclusion:
The gut microbiota has a fundamental influence on host fitness by contributing to host
metabolic capabilities, immunity levels and development. The gut microbial community is in
turn influenced by several factors such as genetics, environment, food intake, geographical
location and other individual differences of the host. This explores the metabolic potential of the
gut micro flora and uncovers functional variation with diet or host associated factors.
This study provides the microbial density in gut of four economically important fishes
(Oreochromis niloticus, Pangasius bocourti, Catla catla and Ictalurus furcatus) of Chikmagalur
town available throughout the year. Comparatively the microbiome of catla showed higher
colony count (5.3×105 CFU/ml), this depicts that the microbiome within gut of catla is more
compared to other selected fishes. Catla catla is comparatively large when compared to other
fresh water fishes and they are voracious feeders feed on many phytoplankton as well as some
zooplanktons. Due to this property of catla there are chances of presence of microbial flora in its
gut in large quantity.
The study has established the basis for future large-scale investigations of the gut
microbiota of fishes. In order to make assumptions about the ecological consequences of
107
microbiome composition, a much deep and large-scale investigation of the gut microbiota of fish
in this region is also necessary.
References:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC614139
http://ajol.info/index.php/njbas/index Nigerian Journal of Basic and Applied Science (2010),
18(2): 217-221.
http://www.ijcmas.com.International Journal of Current Microbiology and Applied Sciences
9(3):1716-1738
https://www.jove.com/v/10507/serial-dilutions-and-plating-microbial-enumeration.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6141390/ Indian J Microbiol. 2018 Dec; 58(4):
397–414.
https://www.researchgate.net/publication/272342365_Bacteria_Associated_with_Fresh_Tilapia_
Fish_Oreochromis_niloticus_Sold_At_Sokoto_Central_Market_in_Sokoto_Nigeria
Muthukumar P. and C. Kandeepan (2015): Int.J.Curr.Microbiol.App.Sci. 4(3): 607-616
108
(ISBN: 978-93-91768-45-4)
STUDIES ON BREEDING CHARACTERISTICS OF NATURALLY AND INDUCED

BRED DOTTYBACK FISH, PSEUDOCHROMIS DILECTUS (LUBBOCK, 1976)
Nanthini devi Karuppaiya*1,2, Santhanam Perumal2 and Perumal Pachiappan2
1C.A.S. in Marine Biology, Faculty of Marine Sciences,
Annamalai University, Parangipettai-608 502, Tamil Nadu, India
2Marine Planktonology & Aquaculture Lab., Department of Marine Science,
School of Marine Sciences, Bharathidasan University,
Tiruchirappalli-620 024, Tamil Nadu, India
*Corresponding author E-mail: [email protected], [email protected]
Abstract:
Present study documents the baseline data on the induced captive bred dottyback fish,
Pseudochromis dilectus vis-a-vis natural bred one. High fecundity rate (1000 - 1200 eggs) was
found in natural breeders when compared to the induced fishes (700 - 900 eggs). There was carce
difference found in parental care, fanning frequency and embryonic development in natural and
induced bred fishes. However, the successful hatching was occurred in natural spawning (90-
95%) which was higher than the induced spawned (70 - 75%) fishes. Larvae Size and growth
was high in natural bred compared with induced bred.
Keywords: P. dilectus; Natural spawning; Induced spawning; Fecundity
Introduction:
Marine ornamental fishes are one of the most popular attractions in worldwide due to
their adaptability to live in confinement. Over the past decades, marine aquarium industry has
undergone a significant transformation and this industry growing day by day due to their high
commercial value (Ajith Kumar et al., 2012). Marine aquarium trades are providing employment
opportunity to coastal people and also serve as a source to earned foreign currencies to many
developing countries (Nanthini et al. 2010). India is endowed with wide variety of marine
ornamental fishes, while our contribution was only 2.5% (US$ 3.8 million) to the total Asian
ornamental fish exports (Dey, 2010). Successful breeding of marine ornamental fishes is a rare
phenomenon compared to fresh water ornamental fishes and only a few marine species have
been raised on commercial scale all over the world. The ability to meet demands for marine
109
ornamentals, utilizing wild-caught fish is decreasing due to stringent regulations on collections

that deplete wild stocks and cause damage to fragile coral reef ecosystems (Dhaneesh et al.,
2012). The long-term sustainable trade of marine ornamentals can be achieved only through
development of culture technologies. In this circumstance, possible alternative is captive
propagation of target species which ultimately resulting in decreased dependence on wild caught
specimens. Hence, it would help to safeguard the coral reef, develop a new source of organisms
for aquarium trade and the way to marine biodiversity conservation and also extend this
technology to coastal community for their livelihood development (Setu et al. 2010).
Hatchery production technology of dottyback fishes were lack. The sunrise dottyback, P.
flavivertex was bred by Olivotto et al. (2006) and Sayadi et al. (2012) studied the effect of
temperature and juvenile density for the development of neon dottyback, P. aldabraensis.
Recently, Sweet (2013) reported the hatchery production technology of sixteen species of
dottybacks. Broodstock maturation and reproduction in captivity is a challenging task in
aquaculture. Many works were deals with induced breeding of fishes using hormones (Leong,
1977). Hormone is used for stimulate the fish to spawn either naturally or artificially and various
combinations of hormones can be used to induce ovulation of eligible female and male fishes.
Few hormones are mostly used in aquaculture industry are human chorionic gonadotropin
(HCG), follicle stimulating hormone (FSH), luteinizing hormone (LH), thyroid stimulating
hormone (TSH), estrogen, testosterone and fish pituitary glands (Dave and Sukumaran, 1984).
Peter (1980) found that HCG and FSH have induced ovulation, when used alone, while Yaron
(1995) recommended HCG as the preferred hormone for Morone culture.
According to Lam (1982), effective dosage of HCG may vary from species to species.
This hormone is currently preferred by most of the aqua-culturists, because it is effective,
economically feasible and readily available in the markets (Marte et al., 1987). Marine fishes are
concern, some species will spawn after simple environmental changes and others required more
advance methods, including administration of hormone products (Cheah and Lee, 2000). In some
tropical freshwater species, sexual maturation takes longer year. In Morulius chrysophekadion
takes 2 to 3 years (Unsrisong et al., 1990), Osphronemus gouramy (Purakkiet et al., 1990),
Probarbus jullieni (Rodrarung and Janesirisak, 1990) and Puntius gonionotus takes more than 10
years (Sukumasavin and Leelapatra, 1993).
In India, studies on hatchery production technology of dottyback fishes are lacking. To
evaluate these strategies, the present work has been planned to develop breeding technology of
red head dottyback, P. dilectus and also an attempt was made with induced breeding, using
Human Chorionic Gonadotropin (HCG) hormone. Furthermore, reproductive behavior, fecundity
110
(ISBN: 978-93-91768-45-4)
rate, fanning frequency, parental care and embryonic development during natural and induced
breeding have been documented. The ultimate goal of this study is to extend the perfected
captive production technology of P. dilectus to the aqua farmers for their livelihood
development.
Collection and Broodstock development of P. dilectus
The fishes were procured from the traders of Mandapam and Tuticorin coastal areas,
Southeast coast of India. They were packed in oxygenated polythene bags, one fish per pocket
containing 500 ml of seawater and they were transported to marine ornamental fish hatchery
located at CAS in Marine Biology (Annamalai University). Latter they were accommodated in
cement tank (5000 L capacity) with contains 3000 L estuarine water. The tank was installed with
biological filter, provided with dead coral pieces, PVC pipes and live rocks (Ajith Kumar et al.,
2012). The fishes were fed twice a day with boiled meat of oyster, shrimp, green mussel, clams
and Acetes at the rate of 5% of their body weight. Uneaten food particles and excreta were
removed by siphoning after 30 mins. of feeding. Approximately 50% of water was exchanged at
three days intervals.
Breeding techniques
The protocol proposed by Olivotto etal. (2006) was adopted for natural spawning. The
pair formation was observed after eight months of rearing. Before spawning, their reproductive
behaviors were documented at regular intervals. After the continuous rearing over a year, natural
spawning was observed. The induced breeding was carried out using human chorionic
gonadotropin hormone (Rottmann et al., 1991). The HCG hormone was purchased from local
medical store. Before injecting the hormone, the length and weight of the male and female fishes
were noted. 300 IU volume of hormone was injected (intramuscular) to male and female fishes
separately using insulin syringe needle. After hormonal injection, the behavioural characters of
male and female fishes were observed and documented at regular intervals.
Spawning, fecundity rate and parental care
The spawning intervals, fecundity rate and parental care were studied (Dhaneesh, 2009).
The fecundity rate was calculated using the formula (%) = Number of eggs in particular
area×Total number of egg clutch area×100. Parental care was mostly found to done by male than
female during this time male fish didn‘t consume food properly. The fanning frequency of male
and female fishes was determined at different time intervals. The embryonic developmental
study was carried out as described by Olivotto et al. (2006). The size of egg clutch and
111
individual egg was measured using micrometer. The hatching duration was found to be varied
between natural and induced spawners. The larvae were transferred to the larval rearing tank
(250 L capacity) containing 200 L of filtered estuarine water. The larvae were fed with
microalgae enriched rotifer and wild plankton from second day onwards. The length and mouth
size of the larvae were measured.
Data analysis
The present study results were analysed using Pearson correlation coefficient statistical
analysis using Origin Software. A probability of 0.05 was used for find out the statistical
difference between the means. The means±S.D. of the data result has been expressed in this
study.
Results:
Total 40 numbers of adult fishes were procured and conditioned in quarantine tanks. The
standardized water quality parameters were followed (Salinity 26 - 28 ppt, Temperature 28 -
30ºC, pH 7.8 - 8.2, Dissolved oxygen 4.5 - 5.5 mg/l, Ammonia 0 - 0.01 ppm and photo period
(12 hrs light: 12 hrs dark). Initially, the average length (3.8 - 4.2 cm) and weight (2.2 - 3 g) of the
fishes were measured. After three months rearing differentiation (Length 4.5 - 5.2 cm; Weight
3.2 - 3.8 g) was observed in male and female fishes and they were kept in ten oval tanks. Among
the ten pairs maintained, three fishes were dead due to bacterial infections and their partners
were used for reproductive biology studies. Then the existing seven pairs were reared by feeding
with polychaetes, boiled prawn, clam, oyster and green mussel. Followed by pair formation was
occurred in three pairs and each pair (Length 5.5 - 5.8 cm; weight 3.8 - 4.4 g) was separated and
kept in 750 L capacity spawning tanks. The fishes exhibited reproductive behavior like pairing
after ten months rearing and the length and weight of the fishes were found to vary from 6.2 -
7cm and 4.5 - 5.6g. Spawning always occurred between 11 am to 1 pm.
The average length (cm) and weight (gm) of the fishes were; 6.3 - 7.2 and 4.8 - 5.6
respectively at time of HCG hormone injection. On the 4th day, more reproductive behaviour was
exhibited and spawning occurred between 12 pm to 1 pm. In natural spawning (NS), the
spherical egg ball was around 2 cm diameter and total number of eggs in a clutch was found to
vary approximately 1000 - 1200 nos. In the case of induced spawning (IS), the spherical egg ball
was around 1.5 cm diameter and total number of eggs in a clutch was found to vary between
approximately 700 - 900 nos. The spawning frequency was noticed between ten to fifteen days
intervals in NS.
112
(ISBN: 978-93-91768-45-4)
Table 1: Captive breeding technology of Pseudochromis species

Pseudochromis Spawning Incubation Larvae Size Country References
species rate period and
Fecundity rate
Pseudochromis 95% 4 days and 1000 4.1 - 4.2 mm Italy Olivotto et al.
flavivertex - 1200 nos. with mouth (2006)
gape 140 - 150
μm
Pseudochromis 90 - 95% 5 to 6 days and 3 - 3.5 mm Brazil Mies et al.
flavivertex 1000 - 1200 nos. with mouth (2014)
Pseudochromis 80 - 85% 4 to 5 days and gape 120 to
aldabraensis 900 - 1000 nos. 130 μm
Pseudochromis 80 - 85% 4 to 5 days and
springeri 800 - 1000 nos.
Pseudochromis 91 - 95% 4 days and 400 - 5.1 - 5.3 mm India Madhu et al.
dilectus 500 eggs with mouth (2016)
gape 150 to
160 μm
Pseudochromis 80 - 85% 4 to 5 days and 2.43±0.31 mm Portugal Araujo et al.
fridmani 1500 - 2000 with mouth (2016)
eggs gape 100 to
120 µm
Pseudochromis Natural NS: 4 days and 3.5 - 3.8 mm India Present Study*
dilectus Spawning 1000 - 1200 nos. with mouth
(NS): 90 - IS: 4 to 6 days gape 120 to
95% and 700 - 900 130µm
Induced nos.
Spawning
(IS): 70 -
75%
No spawning intervals observed in induced breeding pairs, as continuous hormone

injection was made at a week interval to obtain frequent spawning. During incubation, the male
fan the eggs continuously, but female done it occasionally and it showed less parental care. The
maximum and minimum fanning in a day made by the male was 1.62 and 0.37 minutes and in
case of female it was 0.88 and 0.06 minutes. The hatching of larvae normally occurred on 4th day
of incubation. The larvae hatching rate has been noticed high in natural spawning (90 - 95%),
when compared to induced spawning (70 - 75%).
113
The newly hatched larvae were measured to be 3.5 - 3.8 mm with the mouth size of 120
to 130µm, were slender and pigmentation was very light. The eyes, mouth, gut and fin fold were
developed. The larvae were fed with algal enriched rotifer (Brachionus plicatilis) and wild
plankton (Copepod, Sagitta, Mysis, Zoea etc) at a density of 15 - 20 nos/ml. The sixth day larvae
were measured about 5.2 - 5.4 mm and the mouth size was 130 - 140µm. On 11th day, the larvae
were measured at 6.5 - 6.8mm and mouth size was 150 - 160µm. Juvenile stage (Length 2.8 - 3.2
cm; weight 0.8 - 1.5 g) was achieved after reared of three months. The present study result was
compared with earlier reports (Table 1).
The present study has provided baseline data for the hatchery production of P. dilectus,
particularly on broodstock development, spawning and larval rearing. In this study, totally 40
numbers of adult fishes were procured and conditioned in quarantine tanks (Fig. 1). Initially, the
size of the fishes was ranged between 3.8 to 4.2 cm and weight 2.2 to 3 g. The water quality
parameters were standardized and followed (Salinity 26 - 28 ppt, Temperature 28 - 30ºC, pH 7.8
- 8.2, Dissolved oxygen 4.5 - 5.5 mg/lit, Ammonia 0 - 0.01 ppm and photo period (12 hrs light:
12 hrs dark). After rearing three months, differentiation was observed in male and female and
they were kept in ten oval tanks separately and during that time, the fishes attained the size
ranged between 4.5 - 5.2 cm and weight 3.2 - 3.8 g. Among the ten pairs maintained, three fishes
were dead due to bacterial infections and their partners were used for reproductive biology
studies. The existing seven pairs were reared and these fishes were fed with polychaetes, boiled
prawn, clam, oyster and green mussel. Followed, pair formation was occurred in three pairs and
each pair was separated and kept in 750 L capacity spawning tanks (Fig. 2). The length and
weight of the fishes ranged from 5.5 - 5.8 cm and 3.8 - 4.4 g at the time of first spawning.
Figure 1: Conditioning of P. dilectus
114
(ISBN: 978-93-91768-45-4)
Figure 2: Experimental set-up for pair formation and spawning
The fecundity rate, spawning frequency, parental care and fanning frequency were
recorded. In natural spawning (NS), the spherical egg ball was around 2 cm diameter and total
number of eggs in a clutch may vary between approximately 1000 - 1200 nos. In the case of
induced spawning (IS), the spherical egg ball was around 1.5 cm diameter and total number of
eggs in a clutch may vary between approximately 700 - 900 nos only. The spawning frequency
was noticed between ten to fifteen days intervals. No spawning intervals in induced breeding
pairs, as continuous hormone injection was made at a week intervals to obtain frequent
spawning. There were no changes in parental care and fanning frequency in NS and IS fishes.
Mostly the male fish guards and protects the eggs until they hatch. During this period, they were
exhibited two processes, fanning and mouthing. It was also noticed, during this time male was
consumed small amount of food only. During incubation, the male fan the eggs continuously, but
female done it occasionally and it showed less parental care. The maximum and minimum
fanning in a day made by the male at one time was 1.62 and 0.37 minutes respectively and in
female, it was 0.88 and 0.06 minutes respectively.
In the present study, the newly spawned eggs were milky white in color. As the embryo
develops, eggs color turns to black on 2nd to 3rd day and later turns to silver color on 4th day,
which is the indication of hatching. At this stage, the glowing eyes of the developing larvae
inside the egg capsule were clearly visible, when viewed from a short distance. The hatching of
larvae normally occurred on the night of 4th day of incubation. The successful hatching of larvae
was occurred in natural spawning was more (90 - 95%), when compared with induced spawning
(70 - 75%). The fishes not showed any parental care with the fry after they hatched. Newly
hatched larvae were very active and swam near the water surface and the yolk sac was almost
completely reabsorbed. First food was offered the day after hatching.
The newly hatched larvae were measured 3.5 - 3.8 mm and mouth size 100 to 120µm,
were slender and pigmentation was very light. The eyes, mouth, gut and fin fold were developed.
115
The larvae were fed with algal enriched rotifer (Brachionus plicatilis) and wild plankton
(Copepod, Sagitta, Mysis, Zoea etc) at a density of 15 - 20 nos/ml. The sixth day larvae were
measured about 5.2 - 5.4 mm and the mouth size was 130 - 140µm. On 11th day, the larvae were
measured at 6.5 - 6.8mm and mouth size was 150 - 160µm. The body was still transparent (Fig.
3). There was no much variation was observed in larval size, growth and feeding behaviour of
the larvae between NS and IS.
Figure 3: Larval rearing system and different days of newly hatched larvae
Discussion:
In worldwide, overall 21 species of dottybacks were bred commercially (Sweet, 2013).
However, in India there is lack of information on breeding of dottybacks in captivity. Totally 13
species of dottybacks are distributed in India, among them only one species (P. dilectus) was
found in Gulf of Mannar region. The red head dottyback fish in the Southeast coast is a popular
aquarium fish that inhabits exclusively in the Gulf of Mannar region. Since, the collection of reef
fishes in India is entirely forbidden and the import is restricted, the P. dilectus is rarely
encountered in aquarium shops and even if they are available, the prices are very high.
Therefore, presently an attempt has been made in this study through captive breeding and rearing
techniques for this important species. Induced breeding technique was also attempted. The
present study results have been compared with earlier reports: spawning, incubation period,
fecundity rate and larvae size was varied between species level in same genus of Pseudochromis.
In these circumstances, red head dottyback was collected and conditioned in captivity using
estuarine water and an attempt has been made to develop the hatchery production technology.
Since, the time taken for natural spawning of this species was lengthy, a trial on induced
breeding using HCG hormone was also attempted. Previous research findings are evidenced that,
HCG is specific responsible for ovulation process. Research is currently being conducted with
116
(ISBN: 978-93-91768-45-4)
different hormones, but these hormones has given promising result in maturation and enhanced
the brood stock development (Harvey et al. 1993). Induced breeding of fishes was achieved
using HCG hormone in Indian major carps (More et al. 2010), Poecilia sphenops (Sudha, 2012)
and Sea bass, Lates calcarifer (Thirunavukarasu et al. 2004; Hill et al. 2009). Madhu et al.
(2016) has been reported the breeding technique of Pseudochromis dilectus and achieved 95% of
spawning and fecundity rate of 400 to 500 eggs. Compared with this present study achieved 90 -
95% of natural spawning and fecundity rate of 1000 - 1200 eggs. In case of induced spawning
achieved 70 - 75% and fecundity rate was 700 - 900 eggs. Highest spawning and fecundity rates
have been achieved from present study.
Finally, it is concluded that this is the first attempt on natural and induced breeding of P.
dilectus using estuarine water. Larval rearing and juvenile production will be strengthened,
which will achieve the goal of successful captive production P. dilectus in captivity. Moreover,
this technology will be extended to coastal community as livelihood development, which can
indirectly conserve this species in wild.
Acknowledgments:
The authors are grateful to Annamalai University for providing the necessary facilities to
carryout the work and to the University Grants Commission, New Delhi for its financial
assistance through the Rajiv Gandhi National Fellowship Scheme (UGC Ref No. F.14-
2(SC)/2009(SA-III)). K.D expresses deep sense of gratitude to her research mentor, Dr. T.T.
Ajith Kumar, Former Assistant Professor & Senior Research Officer, C.A.S. in Marine Biology,
Faculty of Marine Sciences, Annamalai University, who introduced her to the fascinating world
of marine ornamental fish - aquaculture.
References
Ajith Kumar, T.T., Gopi, M., Dhaneesh, K.V., Vinoth, R., Ghosh, S., Balasubramanian, T., and
Shunmugaraj, T., 2012. Hatchery production of the clownfish, Amphiprion nigripes at
Agatti Island, Lakshadweep, India. J. Environ. Biol. 33, 623-628.
Araujo, A.S.D., Nascimento, S.D.W., Emılia, M.Y., Sathyabama, C., 2012. Temporal dynamics
of reproduction of the neotropical fish, Crenicichla menezesi (Perciformes: Cichlidae). The
Scientific World Journal, 1-10.
Cheah, M.S.H., Lee, C.L., 2000. Induced ovulation of the Australian eel-tailed catfish Neosiluris
ater (Perugia) with ovaprim. Asian Fisheries Science 13, 87-96.
117
Dave, H.S., Sukumaran, T., 1984. Some observations on the use of human chorionic
gonadotropin for induced breeding of Indian and Chinese carps. Souvenir of the seminar
on freshwater fisheries and rural development, Rourkela, Orissa. 126 p.
Dey, V.K., 2010. Ornamental fish trade - Recent trends in Asia. Ornamentals Kerala, Souvenir.
39-45 pp.
Dhaneesh, K.V., 2009. Studies on broodstock development, breeding and larval rearing of clown
fish, Amphiprion percula (Lacepede, 1802) in captivity. M. Phil Thesis, Centre of
Advanced Study in Marine Biology, Faculty of Marine Sciences, Annamalai University,
Tamil Nadu, India.
Dhaneesh, K.V., Ajith Kumar, T.T., Swagat, G., and Balasubramanian, T., 2012. Breeding and
mass scale rearing of clownfish Amphiprion percula: feeding and rearing in brackishwater.
Chinese J. Oceanol. Limnol. 30(4), 528-534.
Lam, T.J., 1982. Applications of endocrinology to fish culture. Canadian Journal of Fisheries and
Aquatic Sciences 39(1), 111-137.
Leong, R., 1977. Maturation and induced spawning of captive pacific mackerel, Scomber
Japonzcus. Fishery Bulletin 75(1), 205-211.
Marte, C.L., Sherwood, N.M., Crim, L.W., Harvey, B., 1987. Induced spawning of maturing
milkfish, Chanos chanos (Forsskal) with gonadotropin releasing hormone (GnRH)
analogues administered in various ways. Aquaculture 60(3), 301-303.
Nanthini, D.K., Dhaneesh, K.V., Ajith Kumar, T.T., Balasubramanian, T., 2010. Hatchery units
for clownfish breeding in coastal region: A new component for marine product export from
India. Seafood Export Journal 5, 17-21.
Olivotto, I., Rollo, A., Sulpizio, R., Avella, M., Tosti, L., Carnevali, O., 2006. Breeding and
rearing the sunrise dottyback, Pseudochromis flavivertex: The importance of live prey
enrichment during larval development. Aquaculture 255, 480-487.
Peter, R.E., 1980. Serum gonadotropin levels in mature male goldfish in response to luteinizing-
hormone releasing hormone (LHRH) and des-Gly10-[DA1a6] LHRH ethylamide.
Canadian Journal of Zoology 58, 1100-1104.
Purakkiet, C., Ratanatriwong, V., Durayab, S., 1990. A preliminary experiment on breeding
black shark (Morulius chrysophekadion) by using suprefact and motillium. Phrae Inland
Fisheries Station, Department of Fisheries. Annual Report. 84-91 pp.
Rodrarung, D., Janesirisak, S., 1990. Induced spawning of earthy pond reared jullien carp.
Nongkai Inland Fisheries Station, Udornthani Inland Fisheries Development Centre, Inland
118
(ISBN: 978-93-91768-45-4)
Fisheries Division, Department of Fisheries. Ministry of Agriculture and Cooperatives,

Annual Report. 114-117 pp.
Rottmann, R.W., Shireman, S.V., Chapman, F.A., 1991. Hormone preparation, dosage
calculation and injection techniques for induced spawning of fish. Southern Regional
Aquaculture Centre Publication. Institute of Food and Agricultural Sciences, University of
Florida.
Sayadi, P., Mohammadizadeh, F., Bahri, A.H., 2012. The effect of temperature and juvenile
density on the development of neon dottyback (Pseudochromis aldabraensis). Broodstock.
Journal of Animal Science Advances 2(12), 941-945.
Setu, S.K., Ajith Kumar, T.T., Balasubramanian, T., Dabbagh, A.R., Keshavarz, M., 2010.
Breeding and rearing of regal damselfish, Neopomacentrus cyanomos (Bleeker, 1856): The
role of green water in larval survival. World Journal of Fish and Marine Sciences 2(6),
551-557.
Sukumasavin, N., Leelapatra, W., 1993. Comparison on the biological activities of gonadotropin
releasing hormone and its analogues in combination with domperidone on the induction of
gonadotropin secretion and spawning in the Thai carp, Puntius gonionotus Bleeker. Thai
Fisheries Gazette 46(6), 511-518.
Sweet, T., 2013. Captive-bred marine aquarium fish species list. Coral Magazine 10(1), 54-57.
Unsrisong, K., Jittakorn, A., Sriwattanawarunyu, Thienchareon, P., 1990. A culture of Morulius
chrysophekadion (Bleeker) spawners in earthy ponds. Chiangmai Inland Fisheries
Research Centre, Inland Fisheries Division, Department of Fisheries. Ministry of
Agriculture and Cooperatives, Annual Report. 86-99 pp.
Yaron, Z., 1995. Endocrine control of gametogenesis and spawning induction in carp.
Aquculture 129, 49-73.
119
UNDERSTANDING GENETIC
Vinda Manjramkar
Department of Zoology
B. N. Bandodkar College of Science (Autonomous), Thane
Genetic is the study of transmission of hereditary characters from parents to offspring‘s,

the physical, morphological characters (traits) observed in organisms called as phenotype. The
traits are coded on genes present on DNA segments, whereas RNA fragments in lower
prokaryotes called as genotypes. Different researchers have given definition for the genetics.
Genetics is a field of science that includes the study of inheritance and genetic variations by
investigating the DNA, genes, genome, chromosome and other components of it. The
phenomenon of inheritance was first explained by Gregor Johann Mendel during the late 18‘s.
As per Mendel‘s finding, ―the traits inherited from parents to their offspring‘s.‖ Some traits are
physical while some are biological. What he called a trait is now known as genes; genes pass
down to consecutive generations. The phenotypes of an individual maybe different then the
genotype, expression of phenotypic characters depend upon the expression of dominant and
recessive genes present in the individual. There are various branches of genetic-Human genetics,
Clinical genetics, Population genetics, Molecular genetics, Cytogenetics, Preimplantation
genetics, Plant genetics, Microbial genetics, archaeogenetic are some areas.
Human Genetic:
Gotra system of Hinduism to prevent inbreeding
Hereditary characters are traits expressed in the family from great ancestors, in Hinduism
it called as Gotra system (“the word gotra denotes the progeny (of a sage) beginning with the
son’s son). The marriages from same gotra are prohibited as they have same (mythological)
ancestry, People within the gotra are regarded as siblings and marrying such a person can lead to
higher chances for the child to get genetically transferred diseases or weak genes or
accumulation of recessive genes. Reason behind this practice (not marrying in same gotra) is the
―Y‖ Chromosome which is expected to be common among all male in same gotra. So, the
woman too carries similar X Chromosome and if married, their offspring may be born with birth
defects, in almost all Hindu families, marriage within the same gotra is not practiced, thus to
prevent inbreeding and completely eliminate all recessive defective genes from the human DNA.
The ancestry is tracked from great sages, it is basically lineage of ―Y‖ chromosome, because it is
120
(ISBN: 978-93-91768-45-4)
standalone chromosome which does not have crossing over, gene exchange, thus passes the
genes from father to son to grandson to great grandson and so on (so paternal lineage is
considered). Gotra system also protects the Y chromosome, also diluting the frequency of
defective genes or eliminating them. The gotras are named as Kashyap, Gautam, Shandilya,
Angirasa, Atri, Bhrigu, Vasishta, Kutsa, Bharadwaj are some of the names. Typically, genes
from the mother and father are shuffled or "cross over" to produce a genetic combination unique
to each offspring. But the Y chromosome does not undergo crossing over, and, as a result, its
genes tend to degenerate, while repetitive DNA sequences accumulate.Y chromosomes are
highly dynamic and have mechanisms to acquire and maintain genes," says Amanda
Larracuente, an assistant professor of biology at Rochester.
With a 30% difference between humans and chimpanzees, the Y chromosome is one of
the fastest-evolving parts of the human genome (Wade, 2010). The Y chromosome was
identified as a sex-determining chromosome by Nettie Stevens at Bryn Mawr College in 1905
during a study of the mealworm (class Insecta) Tenebrio molitor. Stevens proposed that
chromosomes always existed in pairs and that the Y chromosome was the pair of the X
chromosome discovered in 1890 by Hermann Henking.Stevens named the chromosome "Y"
simply to follow on fromHenking‘s ―X‖ alphabetically derived (Bainbridge, 2003; Schwartz,
2009).
The idea that the Y chromosome was named after its similarity in appearance to the letter
"Y" is mistaken. All chromosomes normally appear as an amorphous blob under the microscope
and only take on a well-defined shape during mitosis. This shape is vaguely X-shaped for all
chromosomes. It is entirely coincidental that the Y chromosome, during mitosis, has two very
short branches which can look merged under the microscope and appear as the descender of a Y-
shape.
The genes on the Y chromosome cannot undergo genetic recombination, the ―shuffling‖
of genes that occurs in each generation which helps to eliminate damaging gene mutations.
Deprived of the benefits of recombination, Y chromosomal genes degenerate over time and are
eventually lost from the genome. Thus few functional gene are present on y chromosome like,
TDS testis determining factor, sex-determining region SRY, hypertrichosis, Y-chromosome-
linked diseases are rare.
Animal behaviour to prevent inbreeding
Even the animals do not allow inbreeding between brothers and sister. In elephants the
male calf is distanced away from the matriarchal herd ones he attains sexual maturity. The entire
121
heard is of female and males are solitary. Same is the case with the Lions pride. This means it‘s
genetically coded to follow this phenomenon which is passed over from millions of years. This
behavior brings genetic variations and saves the herd from deleterious effects, thus the survival
of species is benefitted. This is subject of evolutionary genetics. Better progeny is reproduced
from out breeding, also called as out breeding vigor, this brings new variations have better
chances of survival, also diluting the recessive genes ,which is observed in nature, whereas,
inbreeding has deleterious effects with expression of recessive and weak genes (alleles from
same ancestors), in breeding also creates depression termed as ‗inbreeding depression‘.
Human intervened in breeding of some domesticated species.People were improving
plant crops and domesticated animals by selecting desirable traits from individuals for
inbreeding, selective breeding. The example milk gene trait in cattle, muscle trait for beef
production, egg laying in white leghorns, fast development of broiler chicken (white leghorn
variety bred for meat), which is doneby using biological skills, sheep are careful selected to
produce more wool, in vitro, in vivo fertilization. Systematic inbreeding and maintenance of
inbred strains of laboratory mice and rats is of great importance for biomedical research. The
inbreeding guarantees a consistent and uniform animal model for experimental purposes and
enables genetic studies in congenic and knock-out animals. Inbreeding is generally deleterious,
even in flowering plants. Inbreeding is generally deleterious, even in flowering plants. Since
inbreeding raises the risk that bad copies of a gene will be expressed, inbred progeny suffers
from reduced viability.
Care taken by lower organisms to save the progeny
Several organisms perform binary fission. Bacteria, for instance, use it as a way to
reproduce. Bacterial fission entails chromosomal replication, chromosomal segregation, and cell
splitting. The protozoans like amoeba, paramecium, euglena etc., undergo asexual reproduction
as well as sexual reproduction. When conditions are optimal and favorable, they use asexual
mode of reproduction called binary fission it can be called as cloning. The word asexual
describes a reproduction that occurs without involving sex cells (gametes). Instead, the somatic
cells undergo an asexual process that will produce a clone of the parent. So that large number of
progenies is produced, latter in unfavorable conditions some of them can survive. These
organisms do undergo sexual reproduction, Woodruff (1907 published in 1929) claim of keeping
paramecium healthy for 22,000 generations without conjugation (sexual reproduction). If binary
fission continues repeatedly for a longer period of time, Paramecium loses its vigor andare
physiologically depressed, reduces in size, ceases to multiply, degenerates in the organization,
and eventually die, but the clone can be rejuvenated to regain its former vigor by nuclear
122
(ISBN: 978-93-91768-45-4)
arrangement, this is brought about by conjugation. Thus, conjugation is essential for continued
binary fission.In species of Paramecium tetraurelia, the asexual line of clonally aging Paramecia
loses vitality and expires after about 200 fissions if the cells fail to undergo autogamy or
conjugation. Experiments by Smith-Sonneborn, Holmes and Holmes and Gilley and Blackburn
demonstrated that, during clonal aging, DNA damage increases dramatically. When clonally
aged P. tetraurelia are stimulated to undergo meiosis in association with either conjugation or
automixis, the genetic descendants are rejuvenated, and are able to have many more mitotic
binary fission divisions.
Therefore the primitive way asexual reproduction advanced to sexual reproduction,
causing the genetic material to reshuffle by mechanism of crossing over in gametogenesis in
higher organisms, so that they get fair chance of survival.
References:
Bainbridge David (2003): The X in Sex: How the X Chromosome Controls Our Lives, pages 3-
5, 13, Harvard University Press.
Gilley, David; Blackburn, Elizabeth H. (1994): Lack of telomere shortening during senescence in
Paramecium. Proceedings of the National Academy of Sciences of the United States of
America. 91 (5): 1955–1958.
Holmes, George E.; Holmes, Norreen R. (1986): Accumulation of DNA damages in aging
Paramecium tetraurelia. Molecular and General Genetics. 204 (1): 108–114.
doi:10.1007/bf00330196. PMID 3091993. S2CID 11992591.
Schwartz James (2009): In Pursuit of the Gene: From Darwin to DNA, pages 170-172, Harvard
University Press.
Smith-Sonneborn, J. (1979): DNA repair and longevity assurance in Paramecium tetraurelia.
Science. 203 (4385): 1115–1117.
Wade N (2010): Male Chromosome May Evolve Fastest. New York Times
Woodruff L. L. (1929): Thirteen Thousand Generations of Paramecium.Biology Proceedings of
the Society for Experimental Biology and Medicine
123
VERMICOMPOSTING OF RICE STRAW USING EARTHWORM

EUDRILUS EUGENIAE
Sivasankari. B*1, Priyadharshini. R2 and Ganesh. S3
1, 2Department of Biology,
3Department of Physics,
The Gandhigram Rural Institute (Deemed to be University), Gandhigram
Abstract:
Rice straw was collected from Veerakkal village, Athoor (T.K), Dindigul District, and
Tamil Nadu and earthworm species (Eudrilus eugeniae) were collected from as Sakthi
Vermiform, Sanampatti, and Vadipatti (T.K), Madurai District, Tamil Nadu. Rice straw are
collected and shopped into small pieces, it was cured in the open to shade area for 15 days. The
vermibeds were prepared by mixing the rice straw with cured cow dung in 1:1(1 kg of pre
decomposed rice straw and 1 kg of cow dung) ratio. The physical parameters such as pH,
electrical conductivity, temperature, nitrogen, phosphorus, potassium, organic carbon were
analysed once in 15 days upto 60 days.
Keywords: Rice straw, Eudrilus eugeniae, vermicompost
Introduction:
Large quantities of organic wastes produced worldwide have necessitated researchers to
develop appropriate waste recycling technologies in order to protect and preserve the
environment. The burning of huge amounts of organic wastes is a common practice in
developing countries to return complex organic resources back into the soil. These scenarios
were seen especially in rice cultivation where rice straws are chopped and open burnt in the rice
field after harvesting. For example, in Thailand which is a major global exporter of rice 8 to 14
million tonnes of rice straw are burnt in the rice field each year (Edwards et al., 1985). In India,
170 million tonnes rice straw is open burnt each year. Open burning of rice straw is also a
problem in many other countries, recycling this large amount of organic waste in an
environmentally friendly manner helps and contributes significantly towards sustainable
agricultural practices (Reddy and Ohkura, 2004).
124
(ISBN: 978-93-91768-45-4)
Vermicomposting process shows great potential in the degradation of wastes converting

some portion of wastes into earthworm biomass and respiration product and expelling in
remaining wastes as earthworm cast (Reddy and Sreenivasa Raju, 1988). The excreted vermicast
is reported to contain high amounts of mineral, vitamins, plant growth hormones, proteins, and
enzymes (Bano et al., 1987). This implies that feeding rates of different earthworm species may
vary in relation to their morphological differences. The selection of earthworm species with a
high rate of growth, reproduction and vermicast production is required to accelerate waste
breakdown and stabilization (Bansal and Kapoor, 2000). Besides that, identifying earthworm
species that produces vermicast with high plant available nutrient content is important to
produces high quality vermicompost. Among the recommended epigenic earthworm species that
have been widely used in vermicomposting of organic wastes in the topics are Eudrilus eugeniae
(Tripathi and Bhardwaj, 2004).
By using a common feed substrate, the composting efficiency of earthworm species in
producing vermicast with high decomposition rate and nutrient content can be performed.
Therefore, the current research is carried out to evaluate the biological effectiveness (Rate of
reproduction, change in total biomass and rate of decomposition) of earthworms Eudrilus
eugeniae as vermicomposting agent ofagriculutural waste rice straw. Vermicomposting is the
process that involves the oxidation and stabilization of organic matter involving the joint action
of earthworms and microorganisms (Reinecke et al., 1992; Pramanik et al., 2007).) into nutrient
rich plant growth media i.e, vermicast.

Collection of rice straw and earthworms:
Rice straw was collected from Veerakkal village, Athoor (T.K), Dindigul District, and
Tamil Nadu and earthworm species (Eudrilus eugeniae) were collected from as Sakthi
Vermiform, Sanampatti, and Vadipatti (T.K), Madurai District, Tamil Nadu given in plate 1 and
2.
Predecomposition of rice straw:
Rice straw are collected and shopped into small pieces, it was cured in the open to shade
area for 15 days. Water was sprinkled on the waste twice in a day in order to hasten the
predecomposing process. A plastic tray of 45x15x30 cm size was used. Predecomposition of rice
straw in tray was given in plate 3.
125
Plate 1: Collection of sample (Rice straw) Plate 2: Collection of earthworms
Plate 3: Predecomposition of rice straw Plate 4: Predecomposition of rice straw
Preparation of vermibed:
The vermibeds were prepared by mixing the rice straw with cured cow dung in 1:1(1 kg
of pre decomposed rice straw and 1 kg of cow dung) ratio. After 15 days of predecomposition of
rice straw givenin plate 4.
Inoculation of earthworm:
The 15 clitellate Eudrilus eugeniae adult earthworms were inoculated into each of these
trays. The earthworms entered into the media immediately after the inoculation except in the
control tray. These trays were kept undisturbed in shades. Watering was done regularly twice in
a day in order to maintain the temperature and moisture content of the medium during the entire
composting period was 60 days.
Physicochemical analysis of vermicompost:
The vermicomposting was carried out in an environmentally controlled experimental
chamber were temperature of 27±1ºc and the vermibeds were maintained to contain a moisture
level of 70-80%. Various physiochemical parameters such as pH, electrical conductivity, and
total nitrogen (%), total phosphorus (%), total potassium (%) and organic carbon were analysed
using the standard methods.
126
(ISBN: 978-93-91768-45-4)

During vermicomposting process physiochemical parameters were measured, and also
the population of earthworms were counted the results are given in table 1 and 2.
Table 1: physicochemical parameters analyzed on 1st, 15th, 30th , 45th ,60th days
Sr.No. Parameters 1st day 15th day 30th day 45th day 60th day
1. pH 7.4 7.7 7.9 7.0 7.5
2. Temperature 290C 300C 280C 280C 270C
3. Electrical conductivity(ds\m) 1.7 1.9 1.6 2.9 2.2
4. Organic Carbon (%) 23.02 29.03 25.09 20.22 19.10
5. Total Nitrogen (%) 2.10 2.02 2.30 1.61 1.43
6. Total Phosphorous (%) 0.98 0.87 1.52 1.20 1.08
7. Total Potassium (%) 2.19 1.64 2.08 1.10 1.93
Table 2: Enumeration of earthworm’s population after composting

Sr.No. Earthworms Tray I Tray II Tray III Tray IV
1 Egg 75 60 55 62
2 Young ones 150 210 230 250
3 Adult 45 50 54 59
After vercomposting process the final compost is given in plate 5
Plate 5: fine compost
Vermicomposting utilizes earthworms as bioreactors to biodegrade organic waste to

humus. Earthworms have important functions by virtue of their feeding and general behavioral
activities like burrowing, digesting, excreting with decomposing, microorganisms and supporting
127
further decomposition of biodegradable matters. They decompose complex waste materials to

similar forms. The whole process is known as vermicomposting. They modify the soil structure,
fertility, improve plant growth and are important in sustaining productivity Bhiday (1994). The
physicochemical parameters of rice straw during vermicompost on 1st day such as the pH (7.4),
temperature 290C, electrical conductivity 1.7 (ds/m), organic carbon (23.02%), nitrogen (2.10%),
phosphorous (0.98%), potassium (2.19%). During 60th day pH (7.5), temperature 270C, electrical
conductivity 2.2 (ds/m), organic carbon (19.10%), nitrogen (1.43%), phosphorous (1.08%),
potassium (1.93%). Bano et al. (1987) also reported the standardization of agro industrial wastes
for vermicompost practices were the physico chemical parameters 30 day such as pH (7.9),
electrical conductivity (1.6), and nitrogen (2.30), organic carbon (25.09), potassium (2.08),
phosphorous (1.52), C; N ratio (10.91) Edwards (1998) and Reddy (2004), also reported on the
vermicomposting of rice straw substrate and cow dung physico chemical parameters 45 day of
such as pH (7.0), organic carbon (20.22), nitrogen (1.61), potassium (1.10), phosphorous (1.20),
C;N ratio (12.55). In higher content of physico chemical characteristics such as the pH, Ec,
Organic carbon, Na, Ca, K was observed when Eudrilus eugeniae introduced vermicompost. The
vermicomposting (Eudrilus eugeniae) the enumeration of growth rate of worms (Egg, Young
ones, Adult) from 60 days worked vermicompost. The growth rate of earth worms after 60 days
in Tray I (Egg 85, young ones 150, adult 45) was analyzed. The growth rate of earth worms after
60 days in tray II (egg 60, young ones 210, adult 50) was analyzed. The growth rate of earth
worms after 60 days in tray III (egg 55, young ones 230, adult 54) was analyzed. The growth rate
of earth worms after 60 days in tray IV (egg 62, young ones 250, adult 59) was analyzed. In the
high rate young ones has been analyzed in the tray I, II, III, and IV. In the high rate of eggs has
been analyzed in the tray I. In the high rate of adult has been analyzed in the tray IV (Kale and
Bano, 1988).
References:
Bano, K., R.D.Kale, and G.N Gajanan (1987): Culturing of earthworms, Eudrilus eugeniae for
cast production and assessment of worm casts as bio fertilizer, J.Soil, Biol.Ecol., 7(2) :98-
104.
Bansal, S. and Kapoor, K.K. (2000): Vermicomposting of crop residues and cattle dung with
Eudrilus eugeniae – Bio resource Technology 73:95-98.
Bhiday, M.R (1994). Earthworms in agriculture, Indian farming march 1944, 31-33pp.
128
(ISBN: 978-93-91768-45-4)
Edwards, C.A. Dominguez, J. and Neuhauser, E.F. (1998): Growth and reproduction of Perionyx
excavates (Perr.) (Megascolecidae) as factors in organic waste management. Biology and
fertility of soil 27:155-161.
Edwards, C.A., Burrows, I., Fletcher, K.E. and Jones, B.A. (1985): The use of earthworms for
composting farm wastes, Ed. J.K.R. Grasser Amsterdam Elsevier, PP.229-241.
Kale, R.D and Bano, K (1988). Earthworm cultivation and culturing technique for production of
vermicompost. Mysore Journal of Agriculture Science, 2:339-344.
ramanik P, Ghosh GK, Chosal PK, (2007): Changes in organic – C, N, P, K, and Enzyme
activities in vermicompost of biodegradable organic wastes under liming and microbial
inoculants. Bio resours. Technol. 98:2485-2494.
Ravinder Reddy, V. and Sreenivasa Raju, (1988): population growth and biomass production of
Earthworm Eudrilus eugeniae in farmyard manure. Journel Res. ANGRAU, 26(1): 17-19.
Reddy, V.M. and Ohkura, K., (2004): Vermicomposting of rice straw and its effect on sorghum
growth, tropical Ecol, 45: 327-331.
Reinecke, A.J., Vilijen, S.A. and Swayman, R.J. (1992): The suitability of Eudrilus eugeniae for
vermicomposting in Southern Africa in terms of their temperature requirements. Soil. Biol.
Biochem, 24, 1295-1307.
Tripathi, G. and Bhardwaj. P. (2004): Comparative studies on biomass production, life cycles
and composting efficiency of Eisenia fetida (savigny) and Eudrilus eugeniae (Kinberg).
Bio resource Technology 92(3): 275-283.
129
DICHLORVOS MEDIATED BEHAVIOURAL RESPONSESIN FRESH WATER

FISH, CHANNA GACHUA (F. HAMILTON)
P. S. Pawar*1 and R. N. Patil2
Yashwantrao Chavan Institute of Science, Satara 415 001 (MS) India
Sadguru Gadge Maharaj College, Karad 415 124 (MS) India
Abstract:
The quality of water resources are rapidly degrading due to increasing industrialization,
urbanization, use of various fertilizers, insecticides and pesticides.Dichlorvos is widely used to
control variety of domestic, agricultural and garden pests which was used to evaluate their
toxicity in fresh waterfish, Channa gachua. Groups of 10 healthy, well acclimatizedfishes
exposed to the different concentrations of dichlorvos for 96 hrs. As a response against toxicity of
the dichlorvos fish showed remarkable behavioural changes such as rapid jerkymovement,
jumping, hypersensitivity,restlessness, fast opercular and oral movement, excessive mucus
secretion, discoloration of body, shading of scales,enormouslyprotruded and yellow belly,
torsion, inverted hangingand frequent surfacing. So tremendous discharged of the toxic
chemicals into the environment may leads detrimental physiological, biochemical and metabolic
effects on non-target organisms.
Keywords: Pesticide, toxicity, behavior, restlessness, hypersensitivity
Introduction:
All types of pollution has emerged as one of themost significant environmental problem
in recenttimes.Due to industrialization, discharging of effluents from various industries and
pesticides caused serious problems in water bodies and consequently affecting the ichthyofauna
directly or indirectly (Kumar and Pant, 1981). The pervasive use of pesticides in agriculture,
public health and forestry ultimately leads to the contamination of aquatic biotopes posing a
great threat to theaquatic environment (Tripathi and Yadav, 2015).
Pesticide is a biological weapon, which is discovered by the human to kill the insect pests
and to fight against vectors of disease. Nowdays pesticide usages became an indispensable and
integral part of world agriculture and it will not stop but will increase day by day. However, the
intensive use of these chemicals in agriculture and public operations has changed the ecological
balance of many non-target organisms like fishes (Todd and Leeuwen, 2000). Unsafe spraying
130
(ISBN: 978-93-91768-45-4)
and improper handling of the chemical pesticides may cause high risk of the health hazards who
doing the job like groundskeepers, fumigators, gardeners, pet groomers, and farmers. Aquatic
ecosystem is the ultimate sink for agricultural, industrial and domestic runoff.
Dichlorvos (DDVP) is the highly potent organophosphate pesticide used worldwide to
control the varieties of pests and ectoparasites. Fishes are ideal custodians for behavioural assays
of various toxicants stress due to direct contact with the contaminated water and their ecological
relevance in ecosystem systems.Behavioural modification is one of the most sensitive indicators
of environmental stress (Olla et al., 1983). Behavior provides a unique perspective linking
between physiology and ecology of an organism and itsenvironment (Little and Brewer, 2001).
Behavior allows an organism toadjust to external and internal stimuli in order to meet the
struggle for surviving in a changingenvironment. Alterations in fish behavior can also provide
important indices for ecosystem assessment and indicates the deterioration of water quality. So
the present research work was aimed to study the behavioural alterations in fish C. gachua after
exposure to dichlorvos.
Material and Methods:

Material:
Experimental fish: The fish C. gachua was selected for experiment due to its easy availability
and suitability for toxicity testing. Live specimen of C. gachua of size 15 ± 1 cm and weight 50
± 5gm were obtained from Krishna River around Karad city with the help of fisher man. They
were acclimatized in laboratory for 10 days.
Pesticide: Commercially available organophosphorus pesticide dichlorvos was used for present
research work which was brought from Local agro chemist shop.
Methods:
Experimental set up: During experimentation, groups of 10 healthy, well acclimatized fishes
were transferred to the experimental aquarium having different concentrations of dichlorvos. In
each experimental group remarkable behavioural changes were observed in fish during the
experimentation as a response against the toxicity of the pesticides.
Result:
Behaviour of control fish
In aquarium the fish from control group showed normal behaviour. They were active and
swim calmly; they showed normal behaviour with well balancing. Their opercular and oral
movement was quite normal.Secretion of mucous was normal.The scales were intact and skin
131
color was normal.They remained at the bottom of aquarium and occasionally they came to the
surface of water, feeding habit was normal. They were active and sensitive to touch and sound.
Behavioral changes in fish exposed to pesticides
The fishes became restless and rapid, erratic swimming was observed. They showed rapid
jerky movement of body and tried to jump out of water to avoid the toxic medium. Fish became
hypersensitive and showed rapid opercular movement. They were hanging vertically in the
aquarium frequently and become inverted because of loss of equilibrillium, they were lethargic
and sluggish. Discoloration, loss of scales and excessive mucus secretion from the body surface
that giving slimy appearance was noticed. Belly was slightly protruded and became
yellowish.Finally the opercular activity was stopped and fish settled down at the bottom of
aquarium after death. At higher concentrations of the pesticides operculum and mouth remain
opened after death.
Discussion:
Animal behaviour is a response to surrounding habitat. In aquatic habitat the behavioural
changes in fishes indicate the water pollution or unfavorable, undesirable change in water bodies.
Contaminants affect the fish behaviour including locomotion, swimming performance, opercular
and oral movement, feeding, etc. Fishes are sensitive and vulnerable to quick response to the
toxicity.
In present study the control fish, C. gachua showed normal pattern of behaviour. But
after exposed to dichlorvos fishes showed drastic and undesirable changes in their behaviour.
They frequently dashes to the wall and tried to jump out from aquarium. They were restless and
their opercular and oral movement was very fast at the beginning of exposure period. Excessive
secretion of mucus, discoloration of body, shading of the scales was observed. Their belly was
protruded enormously and became yellow and they became inverted and floated near the surface.
They lost their balance and settled down to the bottom. Similar behavioural changes were
observed by Deshpande (2000), when he studied the effect of two pyrethroids i.e. fenvalerate and
cypermethrin at sublethal and lethal concentration against L. rohita. According to him these
changed behaviour might be due to combination of disturbance in physiological, biochemical,
enzymological and hormonal aspects in fishes. Bhilave (2001) reported similar behavioural
changes in fish, C. mrigala exposed to cadmium chloride. According to him the changes in
behaviour might be due to disturbances in biochemical and enzymological aspects in fishes. As
per Halappa and David (2009) C. carpio exposed to chlorpyriphos exhibited disrupted behaviour.
According to them pesticide inhibits the acetylcholinesterase activity which leads in to
accumulation of acetylcholine in cholinergic synapses causing hyper stimulation due to that
behavioural changes occurred in fish. Shrivastav et al. (2010) studied dimethoate induced
132
(ISBN: 978-93-91768-45-4)
behavioural changes in H. fossilis. According to them these behavioural changes were the
symptoms of stress of toxicological nature of the environment. Similar abnormality in
behaviourin fish, C. gariepinus were observed by Lapido et al. (2011) after exposure to paraquat
dichloride. Chavan (2014) observed changed behaviour such as increased opercular movement,
increased surfacing with mouth open, loss of equilibrium, irregular swimming, vertical hanging,
rapid jerks to jumps outside and finally settling down to the bottom, excessive mucus secretion
and yellowness on body in C. mrigala after exposure to lead acetate and mercuric chloride. Dey
and Saha (2016) reported excessive mucous secretion, fast swimming, hyperactivity, increasing
opercular movement, pale gills, irregular, erratic swimming, loss of balance, blood patch on
operculum in fish, L. rohita after exposed to lambda cyhalothrin 5% EC and marshal
(Carbosulfan 25 % EC). C. gachua is an air breathing fish which visited surface of water
periodically to engulf atmospheric oxygen but the surfacing behaviour was found altered by
toxic medium in present study. The increasing surfacing activity observed at lethal concentration
of pesticides, in the present study might be the response of fishes to meet the demand of oxygen
from air because of suffocation and in oxidative or metabolic stress. Surfacing phenomenon i.e.
significant preference to upper layers in exposed group might be due to elevated demands for
oxygen during the exposure period (Katja et al., 2005). Here it is assumed that the dullness in
body color and altered metabolism might be the response by the fish towards the adaption to the
toxic substances. The feeding was impaired and reduced drastically from toxic medium that may
leads into lower energy production that may compel the fish to use of the stored biochemical
constituents to cope up with the high energy demand. Depression in appetite is a common
response that can impact on growth and reproduction of fish. In present study it is assumed that
the organophosphate pesticides might inhibit the acetylcholinesterase (AchE) activity as stated
earlier by Gohel and Dodia (2015) that may paralyze the sensory and neuromuscular system that
leads into changes in normal behaviour and muscular function resulted into the hypersensitivity,
erratic jerky movements, unbalancing, loss of equilibrium in fishes in the present study. On the
exposure to pesticides, fishes showed distress, lethargy and excessive mucus secretion all over
the body. There was excessive mucus secretion by fish which was the response of fish against
the toxic medium that form the barrier between body and toxic medium which may reduce the
contact between body of fish and toxic medium. Similar assumption was mentioned by Rao
(2006) in O. mossambicus exposed to organophosphate insecticides (RPR-V). According to him
mucus forms a barrier between the body and the toxic medium to minimize its irritating effects
or to scavenge it through epidermal mucus.
So, in present work the fish showed well marked abnormal behavioural changes in
response to organophosphate pesticides for the defense and adaptability against the toxicant
133
stressed condition. At the end due to toxic medium fish unable to survive in toxic medium which
results into significant mortality.
Conclusion:
The current study evidenced that dichlorvos is highly toxic and had a detrimental impact
on the behavioural responses of C. gachuaat lethal concentrations. Due to pesticideinstinctive
behavioural response has been reduced and morphological features were affected. Impairments
in behavioural responses might be due to inhibition of brain AchE activity because of action of
organophosphate pesticide. These behavioural responses canbe used as a tool in bio
monitoringprogramme to monitor ecotoxicity risk of pesticides to the test species.
References:
Bhilave, M.P. (2001): Toxicological studies of the heavy metals on the fingerlings of the
freshwater fishes. Thesis submitted to Shivaji University Kolhapur. P 70-75.
Chavan, V. R. (2014): Effect of Heavy Metals on Pituitary Gland and Gonads of fish, Cirrhinus
mrigala. A thesis submitted to Shivaji University, Kolhapur. P. 84.
Deshpande, V. Y. (2000): Effects of synthetic pyrethroids on freshwater fish- Labeorohita.
Thesis submitted to Shivaji University, Kolhapur. 47-50.
Dey, C. and Saha, S. K. (2016): Histological alteration in different tissues of freshwater teleost
Labeorohita (Hamilton) induced by Lambda- cyhalothrin 5% EC and marshal (carbosulfan
25%EC). J. Biol. Today's World. 5 (7): 107-112.
Gohel, P. D. and Dodia, P. P. (2015): Behavioral alteration in Labeorohita exposed to lethal
concentration of cypermethrin (synthetic pyrethroid). Int. J. Rec. Sci. Res. 6 (6): 4818-
4820.
Halappa, R. and David, M. (2009): Behavioural responses of the freshwater fish, Cyprinus
carpio (Linnaeus) following sublethal exposure to chloropyrifos. Turk. J. Fish.Aquat. Sci.9
: 233-238.
Katja, S., Georg, B.O.S., Stephan, P. and Christian, E.W.S. (2005): Impact of PCB mixture
(Araclor 1254) and TBT and a mixture of both on swimming behavior, body growth and
enzymatic biotransformation activities (GST) of carp (Cyprinuscarpio). Aquatic
Toxicology. 71: 49-59.
Kumar and Pant (1981): Histopathologic effect of acute toxic levels of copper and zinc on gills,
liver and kidney of Puntiusconcontius (Ham). In. J. Exp. Biol. 19 – 191.
134
(ISBN: 978-93-91768-45-4)
VERMICOMPOSTING OF SUGAR CANE BAGASSE USING EISENIA FETIDA

Sivasankari B*1, Nathiya G.2 and Ganesh S3
1, 2Department of Biology,
3Department of Physics,
The Gandhigram Rural Institute (Deemed to be University), Gandhigram
Abstract:
Sugar Cane waste was collected from surrounding area of Kodairode, Dindigul (district),
Tamil Nadu. Earthworm species, (Eisenia fedita) were collected from SS Vermiform
Pandiyarajapuram, Vadi Patti, Madurai District, Tamil Nadu India. Sugar Cane waste are
collected and cut into small pieces and kept under shade condition. Water was regularly sprinkled
and the substrate was regularly turned for 15 days. For the preparation the vermicompost the
predecomposed sugar cane waste was directly mixed with cow dung in 1:1 (1 kg of predecompost
and 1 kg of cow dung ) ratio on dry weight basis in tray (15cm x60 cm size ). The substrate
should hold 60-80 percent moisture content and kept for 24hrs stabilization. Fifty numbers of
healthy, cilitellate Eisenia fedita king berg were introduced in the plastic tray. The
vermicomposting trail was carried out in the rearing room. The substrates were turned once in 3
days. The vermicompost was analysed for various physico chemical parameters .such as pH,
electric conductivity, total phosphous, total nitrogen and total potassium using standard
procedure
Keywords: Sugar Cane bagasse, Eisenia fedita, vermicompost
Introduction:
In Indian economy the sugar mill has an essential place and contributes considerably to
its export. India is the second longest producer of suger and its by product amongst the 83 sugar
cane producing countries in the world. The industries of large to medium size as compared to
138 during 1950-1951. About 270 million tons of sugar cane per year is produced in India.
During the manufacturing process large amount of by products such as bagasse press mud and
sugar cane residue are produced. Bagasse is the fibrous waste produced in the sugar cane juice
extraction process Ansari and Ismail (2001). It constituents cellulose (50%), hemicelluloses
135
(25%) and lignin (25%) bagasse is a highly homogenous materials constitute around 30-40% of
pith fibre, which is obtained from the core of the sugar cane. The estimated generation is 0.25-
0.30 per ton of sugar cane. Vermicomposting is a cost effective, ecofriendly and appropriate
waste disposal technique for efficient recycling of animal waste, crop residues, agro industrial
and sewage sludge. Sugarcane trash and press mud could be used for vermicomposting along
with cow dung and other crop residues. Vermicomposting improves the nitrogen and
phosphorous availability besides improving soil physical properties earthworm act as bio-
concentrates of heavy metal and toxic material (Edwards and Bohlen, 1996; Edwards, 2004).
Vermicompost usage improves crop production and material soil health. E.fetida worm are used
for vermicomposting of both domestic and industrial organic waste. They are native to Europe
but have been introduced to every other continent expect Antarctica (Ismail, 1997, 2005).
Materials and methods:

Collection of sample:
Sugar Cane waste was collected from surrounding area of Kodairode, Dindigul (district),
Tamil Nadu, the sugar cane waste was given in plate 1.
Plate 1: Collection of sample Plate 2: Adult earthworm

(sugar cane waste) (Eisenia fedita)
Collection of earthworm:
Earthworm species, (Eisenia fedita) were collected from SS Vermiform
Pandiyarajapuram, Vadi Patti, Madurai District, Tamil Nadu India. The earthworm species
(Eisenia fedita) was given plate 2.
Predecomposition of sugar cane waste:
Sugar Cane waste are collected and cut into small pieces and kept under shade condition.
Water was regularly sprinkled and the substrate was regularly turned for 15 days. The
predecompost tray was given plate 3.
136
(ISBN: 978-93-91768-45-4)
Preparation of vermibed:
For the preparation the vermicompost the predecomposed sugar cane waste was directly
mixed with cow dung in 1:1 (1 kg of predecompost and 1 kg of cow dung) ratio on dry weight
basis in tray (15cm x60 cm size ). The substrate should hold 60-80 percent moisture content and
kept for 24hrs stabilization.
Inoculation of earthworm:
Fifty numbers of healthy, cilitellate Eisenia fedita king berg were introduced in the plastic
tray. The vermicomposting trail was carried out in the rearing room. The substrates were turned
once in 3days.
Physico Chemical analysis of vermicompost:
The vermicompost was analysed for various physico chemical parameters .such as pH,
electric conductivity, total phosphous, total nitrogen and total potassium using standard
procedure (Mane and raker 2012): colorimetric method with molybdenum in sulphuric acid.
Total potassium was determined after digesting the sample in di-acid mixture concentrated
HNO3: concentrated HCl, by flame photometer. After 60th day, the compost was sieved and
sugar cane waste vermicompost were collected.

During vermicomposting process physiochemical parameters were measured, and also
the population of earthworms were counted the results are given in figure 1 and 2.
250
200
electrical conductivity
150 carbon
potassium
nitrogen
phosphorous
Ph
100
50
0
day 1 day 15 day 30 day 45 day 60
Figure 1: physicochemical parameters analyzed on 1st,15th, 30th , 45th ,60th days
137
350
300
250
200 Egg
young ones
adult
150
100
50
0
tray 1 tray2 tray3 tray 4
Figure 2: Enumeration of Earthworms population in vermicompost

Tray 1 –Egg (120), Young ones (300), Adult (250); Tray 2 – Egg (110), Young ones (285), Adult (235);
Tray 3 - Egg (114), Young ones (270), Adult (220); Tray 4 - Egg (90), Young ones (280), Adult (210)
After vermicomposting process the final compost is given in plate 4.
Plate 3: Predecomposition of sugar cane Plate 4: Vermicompost
The physic chemical parameters of sugarcane vermicomposting for I day such as the pH
(7.2), organic carbon (15.3), nitrogen (2.05), phosphorous (0.85), potassium (2.32), C:N ratio
(10.96). Vermicomposting of presented from sugarcane were there physic chemical parameters
in 15 day such as pH (7.3), organic carbon (14.8), nitrogen (2.01), potassium (2.10) phosphorus
(1.85) C:N ratio (14.37), Vermicompost practices were the physic chemical parameter 30 days
such as pH (7.5), electrical conductivity (4.4) and nitrogen (2.02), organic carbon (13.5),
potassium (1.85), phosphorus (1.05), C:N ratio (10.92). Vermicomposting of sugarcane physio
chemical parameter 45 days such as pH (7.0), organic carbon (13.0), nitrogen (1.50), potassium
(1.50), phosphorus (1.50), C:N ratio (12.33). Vermicomposting of sugarcane physico chemical
138
(ISBN: 978-93-91768-45-4)
character 60 days such as the pH (7.1), organic carbon (12.5), nitrogen (1.45) potassium (1.45),
phosphorous (1.39), C:N ratio (13.3). The conclusion of the study was that earthworm Eisenia
fedita is well adapted to sugarcane waste.
In higher content of physico chemical characteristic such as the pH, EC, Organic carbon,
Na, Ca, K was observed when Eisenia fedita introduced vermicomposting Ismail (1995) and
Kale 1998. The vermicomposting (Eisenia fedita) the enumeration of growth rate of worms (egg,
young ones, adult) from 60 days worked at vermicompost. The growth rate of earth worms after
60 days in Tray I (egg 120, young ones 300, adult 250) was analyzed. The growth rate of earth
worms after 60 days in tray II (egg 110, young once 205, adult 235) was analyzed. The growth
rate of earth worm after 60 days in tray III (egg 114, young ones 270, adult 220) was analyzed.
The growth rate of earth worms after 60 days in tray IV (egg 90, young ones 280, adult 210) was
analyzed. In the high rate young ones has been analyzed in the tray I, II, III and IV. In the high
rate of eggs has been analyzed in the tray I. In the high rate of adult has been analyzed in the tray
IV (Talashilkar and Power, 1998; Stoffella and Kahn, 2000).
References:
Ansari, A.A and Ismail, S.A. (2001). Vermitechnology in Organic Solid Waste Management.
Journal of Soil Biology and Ecology 21:21-24
Edwards, C.A. (2004). The Use of Earthworms in the Breakdown and Management of Organic
Wastes. Florida. CRC Press
Edwards, C.A. and Bohlen, P.J. (1996). Biology and Ecology of Earthworms. (3rd ed.),
Chapman and Hall, London. 426p.
Ismail, S.A. (2005). The Earthworm Book. Other India Press, Mapusa, Goa. 101p.
Ismail, S.A. (1997). Vermicology: The Biology of Earthworm. Calcutta: Orient Longman Press,
Hyderabad. 92p.
Ismail, S.A. (1995). Earthworm in soil fertility management. In Organic agriculture (Thampan
ed.). Peekay Tree Crops Development Foundation, Cochin, India. pp:77-100.
Kale, R.D. 1998. Earthworm Cinderella of Organic Farming. Prism Book Pvt Ltd, Banglore,
India. 88p.
Stoffella, P.J. and Kahn, B.A. (2000). Compost Utilization in Horticultural Cropping Systems.
London.
Talashilkar, S.C. and Power, A.G. (1998). Ectocology for pollution control and environmental
management. Enviro Media, Karad
139
BLACK SOLDIER FLY, HERMETIA ILLUCENS L.: A NUTRITIVE INSECT AND A

SOLUTION TO LIVESTOCK FEED
Karuna P. Ganvir
Dr. R. G. Bhoyar Arts, Commerce and Science College, Seloo, Dist. Wardha
Abstract:
The food industries require new supportable sources of protein and fats which can be
received by the use of insects like Hermetia illucens L at an industrial scale. Black soldier fly
larvae have a potential to feed on any organic waste matter and convert it into insect biomass so
it can be utilised as a livestock feed. Livestock sector like poultry, aquaculture, piggery, duck
culture etc. always struggling to fulfil the demands of nutritious feed. Insects like black soldier
fly have been identified as potential alternatives to the predictably used protein sources in
livestock feed due to their rich nutrient content and the fact that they can be reared on organic
wastes streams. Products made from insect biomass resultant from the organic waste streams are
better suitable for industrial large-scale production of insect meal, for this purpose systematic
study of nutritive value of these larvae is needed. Present study confirmed that different nutritive
parameters like crude protein, fats, amino acids, fatty acids, vitamins, flavonoids, minerals etc.
gained by of Black soldier fly larvae fed on different organic wastecan be utilized as an
alternative feed for livestock sector.
Keywords: Black soldier fly, Hermetia illucens L., livestock feed, nutritive value, organic waste,
insect biomass
Introduction:
The food demand will probably increase by 70% by the year 2050 worldwide so as to
meet the demands of 9.7 billion people who are expected to inhibit the world by that time
(Tilman et al., 2011). While the world population continuously growing along with resource
consumption and waste generation, the food resources are becoming limited, causing various
responses from different sectors of society (Paulin et al., 2018). The use of livestocksectors like
poultry, fish, shrimp, piggery etc. utilized as food is the solutionin the demanding and developing
140
(ISBN: 978-93-91768-45-4)
world population which can solve the poverty and provide food security (Thornton et al., 2010;
Otte et al., 2012).
India‘s poultry industry signifies a major success story. Poultry production has been
growing at the rate of around 8 percent per annum, with an annual turnover of US$ 7500 million,
while the agricultural production has been increasing at the rate around 2 percent per annum over
the past two to three decades. In India since 1970s, Universal production, consumption and trade
of poultry meat has grown faster than any other meat. During 1990s demand growth for poultry
meat enhanced when demand growth reducedfor other meats and continued to lead the expansion
of poultry meat trade (Maurice et al.)
As India received its vast coastal area, it offers immense opportunities for fisheries in
both marine and inland waters. India has the second largest share in global aquaculture market,
mainly due to its 2.36 million hectares of ponds and tanks, a coastline of 7,500 kms and brackish
water area of 1.1 million, which offers a great opportunity for fish farming. In India, the fisheries
sector is a means of income for ̴ 28 million people in the country. The Food and Agriculture
Organisation (FAO) 2020 report discovered that Asia has the largest share in the global fish
production at 89%, since the last 20 years. Fish production in the country reached a record high
of 14.16 million metric tonnes between 2019 and 2020.
Poultry farming and aquaculture are the two sectors which can fulfil the demand of
protein diet in future. As these two sectors increasing day by day feed demand for these sectors
willalso increase. Soyabean, corn, fish meal prices are high cannot fulfil the demand of
flourishing livestock farming due to food feed competition. Increase of soybean production in
tropics resulted in land grasping and deforestation in addition to other negative social and
environmental penalties (Foley et al., 2011). For reducing such impacts there is an urgent need
to replace these traditional feed ingredients such as soybean and fish meal with innovative,
economically valuable and environmentally sustainable ones (Tschirner and Kloas, 2017).
Several insect species feed on various types of organic waste streams can be used as an
alternative feed for livestock as these insects gain proteins, fatty acids, micronutrients and gain
high amount of energy (Van Huis et al., 2003; Rumpold et al., 2013; Finke and Oonincx, 2014;
Nowak et al., 2016). The Black Soldier Fly (BSF) Hermetia illucens L. is recognised as
promising alternative source of protein for animal feed (Hale, 1973; Hopley, 2016). The larvae
of this fly can feed on any organic waste like vegetable and Kitchen residues, municipal organic
waste, farm waste, dried distillers‘ grain etc. and reduce the organic waste 50-60% and turn them
into high protein biomass (Craig et al., 1994). Black soldier flies dry weight containsupto 50%
141
crude protein (CP), up to 35% lipids and have amino acid profile that is similar to that of
fishmeal (Elwert et al., 2010). These larvae are discovered as an alternative source of protein for
feed of livestock like poultry, piggery, several types of fishes, shrimps etc.
Black soldier fly (BSF)
Black soldier fly (Hermetiaillucens L) belongs to order Diptera,family Stratiomyidae, life
cycle divided into four phases: eggs, larva, pupa and adult stage. Last larval stage is prepupa
where it migrates from wet food substrate to dry place and converts into pupa. Larval stage lasts
for 14 to 18 days, depending on environmental temperature and pupal stage lasts for about two
weeks. Adult flies emerge form pupal stages prefer to fly towards bright light. Adult flies do not
feed on any food but gravid female flies attract towards smelly food for laying eggs. A single
female can lay 500 to 800 eggs at a time in a bunch in cracks or crevices above the garbage. BSF
larvae reduce the plant or animal waste up to 80%, digest harmful bacteria, adult flies donot
transmit any pathogens from waste food or garbage so these flies can be reared and utilised for
industrial use.
Why Black soldier fly larvae (BSFL) for livestock feed
During meal more than one -third of the edible parts of food gets lost or wasted, insects
play an important role in managing and converting food waste as they can be reared on large
variety of such bio-waste substrates and can be converted into insect biomass (Spranghers et al.,
2017; Cappellozza et al., 2019). Black soldier fly is one of the most effective bio converters
reduce organic waste up to 75% and convert it into biomass havinghigh-quality nutrients like
protein and lipids (Gold et al., 2018).
Zhenghui Gao et al. (2019) fed BSF larvae on maize straw fermented with Aspergillus
oryzae for 24hrs in ratio of 4000:1 and found that larvae contained 41.76% crude protein, 8.24%
crude ash, 30.55% crude fibre, monounsaturated fatty acid 24.86%, polyunsaturated fatty acid
25.37% while saturated fatty acid 45.41% and concluded that the black soldier fly have the
ability to bio convert maize straw and after BSFL bioconversion the maize straw residue can be
used as organic fertilizer.
Andrea Scala et al. (2020) reported that the rearing substrates impacts growth and
macronutrient composition of BSF larvae reared at an industrial scale. Author used three organic
waste streams e.g. banana, appleand spent grainfrom brewery. BSF larvae fed on only fruits diet
as well as on combination of these food diets. BSF larval crude protein content was high i.e.
47.83%, 48.01% and 45.61% when fed on spent grain, Apple +spent grain and banana + spent
grain mixed diet respectively as compared to fed only on fruits. This diet also affected crude
142
(ISBN: 978-93-91768-45-4)
lipid content of BSF larvae. Crude lipid content of larvae fed on fruits i.e., apple, banana and
apple +banana was 36.1%, 27.9% and 33.4% respectively.
Marwas Shumo et al. (2019) took the advantage of easily available organic waste streams
in Kenya to produce nutrient rich black soldier fly larvae. Nutrient value was checked when
larvae were fed on three organic wastes i.e., chicken manure, brewers spent grain and kitchen
waste. Crude protein in larvae fed on these wastes were 41.1%, 33% and 41.3% when fed on
chicken manure, kitchen waste and spent grain respectively. Ether extract of larvae fed on same
waste diet was 30.1%, 34.3% and 31% respectively. Author also evaluated minerals, fatty acids,
amino acids, flavonoids and vitamins and concluded that BSFL can be effectively used as a high-
quality feed as a substitute for other animal or plant derived protein sources in commercial
livestock feed.
The growing demand for fish and soyabean meal in livestock and aquaculture feeding
gives alternative protein carriers to animal feed industry and use of insect-based feeds is
gradually is a matter of interest among fish-feed producers, scientists and policy makers
(Andreas et al., 2014). Different levels of fishmeal replacement by using black soldier fly larvae
meal had been tested on rainbow trout showed neither signs of higher mortalities nor of nutrient
deficiencies observed and concluded that BSF meal can replace fish meal up to alevel of 50% in
trout feeds.
To search alternative sources of poultry feed is still a big concern by nutritional
researchers, by finding balance between high performance and low cost (Khoramabadi et al.,
2014). BSF larvae can be a high value feed as they are rich in protein (37% to 63%) and lipids
(15%-49%) and have better amino acid profile than soybean meal (Barragan-Fonseca et al.,
2017). Schiavone et al. (2017) demonstrated that defatted BSF meals can be excellent source of
apparent metabolizable energy and digestible amino acids for broilers, results into a better
efficient nutrient digestion.
BSFL can replace all the time more expensive protein sources used in poultry,
aquaculture and livestock composite diet formulation, such as fish meal and soybean meal,
which holds the potential to improve future food and feed insecurity. Xiu Liu et al. (2017)
studied metabolic changes in nutrition composition of BSF from egg to adult. Author found a
rapid increase of crude fat content since the development of 4-14 days of larvae with its
maximum level reached 28.4% in dry mass while crude protein reached highest level of 46.2% at
early pupa stage.
143
Conclusion:
It is possible that black soldier fly larvae generated protein and fat could be used as a
replacement for other protein and fat resources in livestock feed industries. Protein and fat
content in larvae very much depends on the type and quality of organic waste fed to the larvae.
By providing different types of organic waste to the BSF larvae we can get essential nutrients
like crude protein, crude lipids, amino acids, fatty acids, vitamin, minerals etc as an insect
biomass and such nutritious insects can be utilised as an alternative feed to the livestock or can
be used its by-products.
References:
Andrea S., Jonathan A., Cammack, Rosanna S., Carmen S., Antonio F., Sabino A. B., Jeffery K.
T., Patrizia F. (2020). Rearing substrate impacts growth and macronutrient composition of
Hermetiaillucens (L.) (Diptera: Stratiomyidae) larvae produced at an industrial scale.
Scientific reports (2020) 10:19448. DOI: https://doi.org/10.1038/s41598-020-76571-8
Andreas S., Stefan W., Ralph N., G Hoerstgen-Schwark, (2014). Black soldier fly
(Hermetiaillucens) larvae-meal as an example for a new feed ingredients class in
aquaculture diets. Proceedings of the 4th ISOFAR Scientific Conference. ‗Building Organic
Bridges‘, at the Organic World Congress 2014, 13-15 Oct., Istanbul, Turkey (eprint ID
24223)
Barragan-Fonseca KB, Dicke M, Van Loon JJA. Nutritional value of the black soldier fly
(Hermetiaillucens L.) and its suitability as animal feed: a review. J Insect Food Feed.
2017;3:105-20.
Cappellozza, S., Leonardi, M.G., Savoldelli, S., Carminati, D., Rizzolo, A., Cortellino, G.,
Terova, G., Moretto, E., Badaile, A., Concheri, G., et al. A first attempt to produce proteins
from insects by means of a circular economy. Animals 2019, 9, 278.
Craig Sheppard, D., Larry Newton, G., Thompson, S. A. & Savage, S. A value-added manure
management system using the black soldier fly. Bioresour. Technol. 50, 275-279 (1994).
Elwert, C., Knips, I. & Katz., P. A novel protein source: maggot meal of the black soldier fly
(Hermetiaillucens) in broiler feed. Tag. Schweine- Geflugelernahrung 140-42 (2010).
Finke, M. D. &Oonincx, D. Chapter 17- Insects as Food for Insectivores. In In Mass Production
of Beneficial Organisms. (eds Juan Morales-Ramos, M.G.R. & Shapiro-Ilan, D.)583-616
(San Diego: Academic Press., 2014).
Foley, J. A. et al. Solutions for a cultivated planet. Nature 478, 337-342 (2011).
144
(ISBN: 978-93-91768-45-4)
Gold, M., Tomberlin, J.K., Diener, S., Zurbrugg, C., Mathys, A. Decomposition of biowaste
macronutrients, microbes, and chemicals in black soldier fly larval treatment: A review.
Waste Manag. 2018, 82, 302-318.
Hale, O. M. Dried Hermetiaillicens larvae (Diptera: Stratiomydae) as a feed additive for poultry.
J. Ga. Entomol. Soc. 8, 16-20 (1973).
Hopley, D. The evaluation of the potential of Tenebrio molitor, Zophobasmorio, Naophoeta,
Blapticadubia, Gromphardhinaportentosa, Periplaneta americana, Blatta lateralis,
Oxyhalaoduesta and Hermetiaillucens for use in poultry feeds. (Stellenbosch University,
2016)
https://www.ibef.org/research/case-study/opportunities-in-the-indian-aquaculture-industry
https://www.researchgate.net/publication/246457812_India''s_Poultry_Sector_Development_and
_Prospects
Khoramabadi V, Akbari MR, Khajali F, Noorani H, Rahmatnejad E. Influence of xylanase and
vitamin A in wheat-based diet on performance, nutrients digestibility, small intestinal
morphology and digesta viscosity in broiler chickens. Acta Scientiarum. Anim Sci.
2014;6:379-84.
Marwa S, Isaac M.O., Fathiya M.K., Chrysantus M.T., Komi K.M.F., Sevgan s., Sunday E.,
Arnold V.H., Christian B. (2019). The Nutritive value of black soldier fly larvae reared on
common organic waste streams in Kenya. Scientific Reports (2019) 9:10110. DOI:
https://doi.org/10.1038/s41598-019-46603-z
Maurice Landes, Suresh Persaud, and John Dyck. India‘s Poultry Sector: Development and
Prospects.Market and Trade Economics Division, Economic Research Service, U.S.
Department of Agriculture, Agriculture and Trade Report WRS-04-03.
Nowak, V., Persijn, D., Rittenschober, D. &Charrondiere, U. R. Review of food composition
data for edible insects. Food Chem. 193, 39-46 (2016)
Otte, J. et al. Livestock sector development for poverty reduction: an economic and policy
perspective livestock’s many virtues. (FAO, 2012)
Paulin N., Janaina M. K., Christian K.T., Claudine T.T., Jislaire Y, Boniface C. and Theophile
F., (2018). Black soldier flies (Hermetiaillucens Linnaeus) as recyclers of organic waste
and possible livestock feed. Int. J. Biol. Chem. Sci 12(5): 2004-2015, October 2018. DOI:
https://dx.doi.org/10.4314/ijbcs.v12i5.4
Rumpold, B. A. & Schluter, O. K. Nutritional composition and safety aspects of edible insects.
Mol. Nutr. Food Res. 57, 802-823 (2013).
145
Schiavone A, De Marco M, Martinez S, Dabbou S, Reena M, Madrid J, et al. Nutritional value

of a partially defatted and a highly defatted black soldier fly larvae (Hermetiaillucens L)
meal for broiler chickens: apparent ileal amino acid digestibility. J Anim Sci Biotechnol.
2017b;8:897-905.
Spranghers, T., Ottoboni, M., Klootwijk, C., Ovyn, A., Deboosere, S., De Meulenaer, B.,
Michiels, J., Eeckhout, M., De Clercq, P., De Smet, S. Nutritional composition of black
soldier fly (Hermetiaillucens) prepupae reared on different organic waste substrates. J. Sci.
Food Agric. 2017, 97, 2594-2600.
Thornton, P. K. Livestock production: recent trends, future prospects. Philos. Trans. R. Soc. B
Biol. Sci. 365, 2853-2867 (2010).
Tilman, D., Balzer, C., Hill, J. &Befort, B. L. Global food demand and the sustainable
intensification of agriculture. Proc. Natl. Acad. Sci. 108, 20260 (2011).
Tschirner, M. &Kloas, W. Increasing the Sustainability of Aquaculture Systems: Insects as
Alternative Portein Source for Fish Diets. GAIA-Ecol. Perspect, Sci. Soc. 26, 332-340
(2017).
Van Huis, A. &Tomberlin, J. K. Chapter 1. The potential of insects as food and feed. In Insects
as food and feed: from production to consumption. (eds Van Huis, A. &Tomberlin, J. K.)
25-58. (Wageningen Academic Publishers).
Xiu Liu, Xuan C., Hui W., Qinqin Y., Kashif R., Wu Li, Minmin C., Qing Li, Lorenzo M., Jibin
Z., Longyu Z. (2017) Dynamic changes of nutrient composition throughout the entire life
cycle of black soldier fly. Research Article.
Zhenghui G., Wanqiang W., Xiaoheng L., Fen Z., Wen L., Xiaoping W., Chaoliang L.
Bioconversion performance and life table of black soldier fly (Hermetia illucens) on
fermented maize straw. J. of Cleaner Production 230 (2019) 974-980
146
(ISBN: 978-93-91768-45-4)
AN ASSESSMENT OF INVERTEBRATE SPECIES IN WETLAND ECOSYSTEMS

Wasudha J. Meshram
Jagat Arts, Commerce and I. H. P. Science College,
Goregaon -441801, Distt. Gondia, (Maharashtra)
Abstract:
The inland water resource on the surface of earth such as Rivers, Lakes, Reservoirs and
Ponds became the focus of special attention in the early stage of development of science of
ecology. The sources and the nature of fresh water, its motion and changing condition as it flows
to the sea and the life it support to all aquatic living things, are now the subject of Limnology,
the science of fresh water and Hydrobiology the science of aquatic life. Anthropogenic activities
of human beings such as bathing, cleaning, household activities, washing of the livestock, will
increase the nutrient load. This may be stopped and some other alternative may be made for this
to reduce the nutrient loads. So there is need of hour for monitoring the quality and status of
wetland ecosystems in order to check the quality of water. The impact of human activities on
these aquatic ecosystems and to improve freshwater ecosystems conservations should be given
emphasis on these aspects.
Introduction:
The inland water resource on the surface of earth such as Rivers, Lakes, Reservoirs and
Ponds became the focus of special attention on the conservation of wetland ecosystems. The
sources and the nature of fresh water, its motion and changing condition as it flows to the sea and
the life it support along the way, are now the subject of Limnology, the science of fresh water
and Hydrobiology the science of aquatic life.
The hydrobiological studies of Indian waters in relation to the seasonal fluctuation of
water characteristics and biological conditions of lakes, reservoirs, tanks and ponds are scanty in
comparison to the vast area of inland freshwater available. The study of seasonal condition of
pond life in Punjab by Prasad (1916) was perhaps the first attempt towards the study of aquatic
ecosystem in India. Later on, Pruthi (1933) studied seasonal fluctuations physical and chemical
conditions of a tank located in Indian Museum Compound, Kolkatta while Gonzalves and Joshi
147
(1946) tried to correlate abiotic and biotic factors of a tank at Bandra (Mumbai). Fresh water
ecology is an intriguing field because of the great diversity of aquatic habitats.
The wetland ecosystems carrying great loads of biodiversity as it is very much rich in its
nutritional value and productivity (Gibbs, 1993; Paracuellos, 2006) and are one of the
ecologically important method of conservation concern sites owing to its tropic dynamics.
Wetlands are generally less than 3 m deep over most of their area which are usually rich in
nutrients and have abundant growth of aquatic macrophytes. They consists of high densities and
diverse fauna mainly planktons, macrophytes, macro invertebrates which have high value for
biodiversity conservation. Excessive growth of macrophytes in wetlands affects the water quality
adversely and interferes with the utilization of the water body. However, marginal aquatic
vegetation is desirable as it controls erosion, beneficial for wildlife habitat and also supports to
improve water quality of aquatic ecosystems. Aquatic ecosystems are variously studied for the
physico-chemical and biological aspects for their conservation and management.
In an aquatic ecosystem, the physico-chemical environment has profound influence on its
biotic components, as it controls its diversity, biomass and spatial distribution in time and space.
The physico-chemical parameters exert their influence both individually and collectively and
their interaction produces an abiotic environment which ultimately conditions the origin,
development and finally the succession of the biotic communities. Fluctuations in physico-
chemical parameters produce adverse effects on the biotic communities which limit their
production and reduce their ability to compete with other population within the environment. All
aspects of biological heterogenicity i.e. structural, functional or taxonomic were included in the
scope of biodiversity (Sarkar, 2008).
According to the study on the biological diversity, that means the variability among the
living organisms from all sources, including terrestrial, marine and other aquatic ecosystems and
the ecological complexities of which they are the important part; which includes diversity within
species, between species and of ecosystems. The physical and chemical characteristics of water
affect the abundance, species composition, their diversity, stability and productivity of the
indigenous population of aquatic organisms. The biological methods used for assessing quality
of water include collection, counting and identification of aquatic organisms means quantitative
and qualitative analysis, biomass measurement etc. and are very essential for its effective
management and conservation of the wetland ecosystems.
Planktons:
Planktonic organisms are found in all aquatic ecosystems except in torrential rivers. They
are small plants and animals whose powers of locomotion are so limited that they cannot
148
(ISBN: 978-93-91768-45-4)
overcome currents in their ecosystem. Most plankters (phytoplankton and zooplanktons) can
move a bit, however, either, to control their vertical distribution or to seize prey.
Phytoplankton:
Among biotic communities phytoplankton constitute the first stage in tropic level by
virtue of their capacity to convert radiant energy into the biological energy through
photosynthetic activity. Also referred to as primary productivity, the magnitude of
photosynthetic energy fixation depends primarily on diversity and biomass of phytoplankton.
The planktonic photosynthesis play a key role in conditioning the microclimate as it helps in
regulating the atmospheric level of oxygen and carbon dioxide. Wanganeo (2007) stated that
large amount of algae can adversely affect the quality of water by inhibiting the photosynthesis
process for other plants and by reducing dissolved oxygen levels. Thus, excessive amounts of
algae usually lead to poor quality. A major contribution to unwanted algal species growth is the
influx of nutrients such as nitrates, phosphates, sulphates etc from non-point sources.
Apart from primary production, phytoplankton also plays an important role as food for
herbivorous animals. They are also considered as biological indicators of water quality in
pollution studies. To summarize, because of their definite role in cycling of energy and matter in
an ecosystem, evaluation of phytoplankton population in term of their diversity, density,
biomass, spatial and temporal distribution, periodicity and productivity and population turnover,
is of vital significance in the management of an ecosystem.
Phytoplankton constitutes the very basis of nutritional cycle of an aquatic ecosystem.
The maintenance of a healthy aquatic ecosystem depends on the abiotic or physical properties of
water and the biological diversity of the ecosystem (Harikrishanan et al., 1999). The planktonic
study is a very useful tool in understanding the basic nature and general economy of the lake
(Pawar et al., 2006). The phytoplankton belonged to Myxophyceae, Bacillariophyceae,
Chlorophyceae and Euglenophyceae.
Zooplanktons:
Zooplanktons are important in fresh water ecosystem as they indirectly convert the food
energy due to their role as preys of economically important fishes. The zooplankton because of
their short life period respond quickly, but also because of their small size and often their great
numbers are also useful in determining the origin or recent history of given water mass (APHA,
1980). The physical and chemical factors play important role, which for the most part are
responsible for the distribution of the animal life in fresh water habitat. The seasonal cycle of
149
zooplankton are affected by the size, distribution of algae, bacterial prey and pressure of
predatory fishes (Kalff, 2002; Chavan et al., 2006).
Zooplankton diversity is one of the most important ecological parameters in water quality
assessment. The zooplankton study has been a fascinating subject for a long time. Water bodies
rich in phytoplankton are also rich in zooplankton diversity and biomass. Vijaykumar (1999)
stated that in an aquatic ecosystem, zooplanktons play an important role not only in converting
plant food into animal food but also provide an important food source for other higher organisms
including fish.
Zooplankton belonged to Rotifers, Copepodes, Cladocerans and Ostracodes. Biological
assessment of the water quality provides a number of advantages over chemical assessment.
Among fresh water communities, zooplankton and phytoplankton are most common living
organisms and many of them are the bioindicator representatives of pollution. However, the
benthos and macrophytes also helps to show the trophic status of the ecosystem.
Benthic Macro-invertebrate:
The community of organisms living on the bottom of a water body is called the benthos.
It includes benthic macro invertebrate communities of Nematodes, Oligocheates, Insects and
Gastropodes. The benthic macro invertebrates can be used as the barometer of overall
biodiversity in the aquatic ecosystem. The invertebrate communities respond to change in water
quality, integrates impact over a period of times and presence or absence of species can be an
indication of specific environmental condition (Anitha et al., 2004).
The benthic macroinvertebrate biological communities are most frequently used to
evaluate water quality in aquatic environments, and occupy variety of trophic levels, acting on
the nutrients, bottom detritus and water column dynamics (Rodrigues and Capitulo, 2002). The
benthic macroinvertebrates belonged to Nematoda, Annelida, Insecta and Gastropoda. Benthos
are important in the secondary productivity of fresh water ponds and lakes, and playing
significant role in exchange of autochthonous and allochthonous material in an ecosystem. It
has a great ecological importance because they form the food of fishes and their productivity
plays a significant role in sustaining food chain and web. Further the qualitative studies of the
benthic macroinvertebrates have also been stressed by many workers (Naidu, 2005; Sharma et al.,
2007; Sisodia, 2007).
Macrophytes:
The macrophytes stimulate the growth of phytoplankton and help in the recycling of
organic matter. The submerged species of macrophytes at the margin also act as green manure
favouring the occurrence and abundance of zooplankton and benthic fauna. The macrophytes
150
(ISBN: 978-93-91768-45-4)
also provide suitable breeding and sheltering place for macroinvertebrates and fishes and other
aquatic organisms (Meshram, 2003).
Narayana et al. (2007) considered that Vallisneria spp. and Ceratophyllum spp., Chara
vulgaris are the species that preferably grow at unpolluted pond. Among the macrophytes, free
floating, submerged, marginal and Emergent are the different types of varieties of aquatic weeds.
Some macrophytes grow rapidly in summer season. When growth slows down the algae occupy
progressively the surface of macrophytes. Water chemistry especially nutrient concentration,
influence the colonization and species composition of periphyton communities (Charachlis et al.,
1990; Guasch et al., 1995). The biodiversity of Macrophytes has been intensively studied by
various workers.
Periphyton:
The assemblage of organism growing on the submerged object such as plants pebbles and
animal shells etc are called ‗aufwuchs‘ or periphyton. The term periphyton (synonym with
haptobenthos) was first used by Behning in 1924 for the plant growth on Buoys, Ships and
moorings of river Volga. Haptobenthos have much assumed considerable importance as these
organisms occur in appreciable and sufficient abundance in some situations and have found to
form the food of some fishes and other aquatic animals. Periphyton belonged to Cyanophyceae,
Chlorophyceae, Bacillariophyceae and Euglenophyceae and Protozoa, Rotifera, Cladocera,
Ostracoda, Gastrotricha, Oligochaeta, Nematoda and Mollusca with reference to the macrophytes
studied such as Eichhornia, Pistia, Lemna minor, Hydrilla, Azolla, Potamogeton and
Nymphoides species. The study of haptobenthos or periphyton has received very little attention
in India.
Summary:
Wetland ecosystems such as ponds and lakes are considered as natural ecological
resources. The management of such wetland ecosystems has become the need of the hours as
community activity and other sources of pollution as enumerated which can alter limnological
characteristics. Desilting of the pond may be emphasized on some alternatives or solutions could
be worked out in collaboration with local municipality for diversion of untreated domestic
sewage and slaughterhouse wastes and means to restore recreational activities in the wetland
ecosystems.
151
References:
Anitha G., Kodarkar, M.S., Chandrasekhar, S.B.A. and Grace Nalini (2004): Studies on micro-
zoobenthos, Mir Atam Lake Hyderabad, Andhra Pradesh. J. Aqua. Biol. 19(1). 61-68.
APHA, 1980). APHA (1980): Standard Methods for the examination of water and wastewater,
17th Ed. American Public Health Association, Washington D.C.: 1131-1138.
Characklis, W.G., Mcfeters, G.A., and Marshall, K.C. (1990): Physiological Ecology of
Periphyton. Periphyton (Eds. W.G.Characklis and K.C. Marshall). John Wiley and Sons,
New York. 341-394.
Chavan, R.J., Mohekar, A.D., and Hiware, C.J. (2006) : Ecology and Behaviour of Zooplanktons
in Manjara Project Reservoir, Beed in Maharashtra, Advances in Limnology by V.B.
Sakhare, Daya Publication House, Delhi : 151-162.
Devi, Sarla, B. 1997: Present status, potentialities, management and economics of fisheries of
two minor reservoirs of Hyderabad. Ph.D. Thesis, Osmania University, Hyderabad.
Gibbs, J.P. (1993): The importance of small wetlands for the persistence of local populations of
wetlands-associated animals. Wetlands, 13:25-31.
Gonzalvis, E.A. and Joshi, D.R. (1946): Freshwater Algae near Bombay. J. Bom. Nat. Hist. Soc.
46: 154-176.
Guasch, H. and Sabater, S. (1995). J. Phycol., 31, 727.
Harikrishanan, K., Sabu Thomus, Sanil George, Paul Murugan, R., Sathish Mundayoor and Das,
M.R. (1999): A study on the distribution and ecology of phytoplankton in the Kuttanad
wetland ecosystem, Kerala. Poll. Res. 18 (3): 261-269.
Naidu, V.K. (2005) : Fauna of India, Aquatic Oligochaeta, ZSI- Kolkata.
Narayana Raj, Saxena, K.K., and Chouhan, S. (2007): Limnological investigations of Texi
temple pond, Dist. Etawah (U.P.) J. Envi. Biol. 28 (1): 155-157.
Kalff, Jacob (2002): Limnology, prentice Hall Publication Upper Saddle River, New Jersey
7458.
Meshram, (2003): limnological studies of Wadali lake, Amaravati, Ph.D. Thesis.
Paracuellos, M. (2006): How can habitat selection affect the use of a wetland complex by
waterbirds? Biodiversity and Conservation, 15: 4569-4582
Pawar, S.K., Pulle, J.S. and Shendge, K.M. (2006): The study on phytoplankton of Pethwadaj
Dam, Taluka Kandhar, Distt. Nanded, M.S. J. Aqua Biol., 21 (1): 1-6.
Prasad, B.N. and Manjula, S. (1916): Ecological study of blue green algae in river Gomti. Indian
J. Environ. Hlth, 22: 151-168.
152
(ISBN: 978-93-91768-45-4)
Pruthi, H.S. (1983): Studies on the bionomics of fresh water in India. I. Seasonal changes in the
physical and chemical condition of the water tank in the Indian Museum Compound.
Internat. Rev. Ges. Hydrobiol. Hydrography, 28: 46-47.
Rodrigues and Capitulo, (2002): The freshwater crabs of America. Family Trichodactylidae and
supplement to the family Pseudothelphusidae. Faune tropcale XXXI, OROSTOM. Paris:
189.
Sarkar, S. (2008): Norms and the Conservation of Biodiversity, Resonance, and Indian Academ
y of Sciences. 13 (7): 627-637.
Sharma, Madhu Sudan, Sumitra, Meena, Sharma, Vipul, Malara, Heena, and Sharma, Riddhi
(2007): Eutrophication process in an urban lake system of Udaipur, NSL – 2007, Jaipur:
31-34.
Sisodia Sapna, Sudha Singh, C. Padmakar, Mogali J. Nandan and Yadava, R.N. (2007): Seasonal
variation and species diversity of macrozoobenthic communities of a tropical lake, Bhopal.
NSL-2007: 211-214.
Vijaykumar, K. Holkar Devendra and Kaur, Kuldeep (1999) : Limnological Studies on
Chandrampalli reservoir, Gulbarga. Freshwater ecosystem of India by Vijaykumar, K.,
Daya Pub., Delhi., : 59-108 pp.
Wanganeo Ashwani (2007): Plankton: Its uses for the benefit of mankind and its utility in water
management. NSL-2007: 43-50.
153

Advances in Animal Science Volume II

Uploaded by

Copyright:

Available Formats

Advances in Animal Science Volume II

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Advances in Animal Science Volume II

Uploaded by

Copyright:

Available Formats

Advances in Animal Science

Dr. Gyanendra Kumar Dr. Anil Madhukarrao Khole

National P. G. College, B. Raghunath Arts, Commerce

Lucknow (U.P.) and Science College, Parbhani (M.S.)

Mr. Debajyoti Pradhan Dr. Naveen Kumar Chourasia

Egra Sarada Shashi Bhusan College Govt. College Bichhua,

(under Vidyasagar University), Chhindwara

Egra (W. B.)

 Copyright reserved by the publishers

of basic and applied animal science.

with the goal of helping people achieve that feeling of accomplishment.

taking pains in bringing out the book.

Sr. No. Book Chapter and Author(s) Page no.

1. FORMULATION OF FISH FEED USING TRADITIONAL MEDICINAL 1 – 12

2. POTENTIAL OF WHEATGRASS BASED FORMULATED FEED IN 13 – 29

3. PRODUCTION OF VACCINE AGAINST GASTROINTESTINAL 30 – 36

4. TECHNOLOGICAL ADVANCES AND ANIMAL HEALTH 37 – 42

5. CLONING AND GENETIC ENGINEERING 43 – 49

6. EMERGING TOOLS FOR CONSERVATION AND 50 – 54

7. DIMETHOATE: KILLER PESTICIDE 55 – 56

8. DIVERSITY OF MOTHS (LEPIDOPTERA: HETEROCERA) AND 57 – 62

10. SERICULTURE BASED INTEGRATED FARMING SYSTEM FOR 69 – 76

11. AN OVERVIEW OF DIFFERENT TYPES OF TOXIC 77 – 89

12. HEAT STRESS AND ITS IMPACT ON HORMONAL PHYSIOLOGY 90 – 99

13. DETERMINATION OF MICROBIAL GUT DENSITY IN 100 – 108

14. STUDIES ON BREEDING CHARACTERISTICS OF NATURALLY AND 109 – 121

15. UNDERSTANDING GENETIC 122 – 123

16. VERMICOMPOSTING OF RICE STRAW USING EARTHWORM 124 – 129

18. VERMICOMPOSTING OF SUGAR CANE BAGASSE USING 135 – 139

20. AN ASSESSMENT OF INVERTEBRATE SPECIES IN 147 – 153

FORMULATION OF FISH FEED USING TRADITIONAL MEDICINAL PLANT

Materials and method:

Plate I: Pathogens culture

Clinical Sign and Gross Pathology:

Plate III: Dried V. zizanioides

Plate IV: Herbal feed

mossambicus against A. hydrophila infection. According to Winkaler et al. (2007), A. indica

Shariff, M. and Subasinghe, R. (1994). Experimental induction of epizootic ulcerative syndrome

POTENTIAL OF WHEATGRASS BASED FORMULATED FEED IN FRESH

Feeding fish is labor-intensive and expensive. Feeding frequency is dependent on labor

Review of Earlier studies

farming in Bangladesh. Poster Published by Department of Aquaculture, Bangladesh

Mamman T., Ipinjolu J., Magawata I. (2013).Hematological indices of Clarias

Plaza I., García J. L., Villarroel M. (2018).Effect of spirulina (Arthrospira platensis)

PRODUCTION OF VACCINE AGAINST GASTROINTESTINAL NEMATODE

Materials and Methods:

polygyrus) after 1, 5 and 13th day of administration of larval somatic antigens:

polygyrus) after 1, 5 and 13th day of administration of adult somatic antigens:

polygyrus) after 1, 5 and 13th day of administration of larval ES antigens:

polygyrus) after 1, 5 and 13th day of administration of adult ES antigens:

Observations and Results:

Summary and Conclusion:

I-NV-C-2 Infected Non Vaccinated Control-2

I-V-AES Infected and Vaccinated with adult ES Antigens

After 1, 5 and 13 days of administration of larval ES antigens, the protection achieved

TECHNOLOGICAL ADVANCES AND ANIMAL HEALTH

Modern technologies in animal science use:

CLONING AND GENETIC ENGINEERING

EMERGING TOOLS FOR CONSERVATION AND

DIMETHOATE: KILLER PESTICIDE