Biological Water Quality Parameters

CE5170 – PHYSICOCHEMICAL PROCESSES FOR
WATER AND WASTEWATER TREATMENT
Biological Water Quality

Parameters
BIOLOGICAL WATER QUALITY
PARAMETERS
 Aquatic organisms range in size and complexity
 Species Diversity Index → parameter for lakes and
streams
 Pathogens
→ Capable of infecting or transmitting diseases

→ Requires an animal host
→ Bacteria, Viruses, Protozoa and Helminths
 Non-Pathogens
→ Pathogen indicators
→ Analysis of pathogens time consuming
THE MICROBIAL WORLD: SIZES OF MICROBES
Bacterium
0.2-3 µm Protozoan Parasite
Cryptosporidium parvum
~5 µm
Virus
20-80 nm Fungi Parasite
(0.02-0.08 µm) Candida albicans
Larger than 2 µm
PARAMETERS
Bacteria
→ Unicellular organisms
→ Bacilli (Rod shaped)
→ Cocci (Spherical)
→ Spirilla (Spiral)
* Gastrointestinal disorders - symptom
* Example - Vibrio comma; Salmonella typhosa, etc.
→Growth conditions and environmental conditions
may be different for different bacteria
→ Example – Psychrophilic (0 to 20oC), Mesophilic (20
to 45oC) and Thermophilic (> 50oC)
Bacteria’s morphology
PARAMETERS
Viruses
 Simplest includes nucleic acids (DNA or RNA) and protein capsid
 Absolute parasites
 Various disease with symptoms of fever, vomiting, jaundice.
 Size of viruses between 0.02-0.08 µm
 Conc. of viruses in raw wastewater 103-104/100 ml, with infectious

dose 1-10
 Penetration of viruses to groundwater is more likely than bacteria
due to their size, and also due to the extreme high numbers in
infected individuals (infected person – 105 – 1011 viruses per gram
of feces)
 Viruses feature longer survival time than bacteria and more
resistant to disinfection
http://techalive.mtu.edu/meec/module02/Viruses.htm
http://health.howstuffworks.com/medicine/modern/light-virus.htm
HUMAN ENTERIC VIRUSES
 More than 100 types of enteric viruses capable of
causing infection and excreted by humans
 Enteric viruses multiply in the intestinal tract and are
released in fecal matter of infected persons
 Most important human enteric viruses are: polio,
coxsackie, Norwalk virus, rotavirus, adenovirus and
hepatitis A virus.
 The pathogen virus HIV is not waterborne
POLIO VIRUS-POLIOMYELITIS
 50 years ago one of the most feared
diseases
 Symptoms are like flu, slight fever,
headache, sore throat and up to deep
muscle pain and paralysis
 Transmission fecal to oral route
 Found in wastewater and water

ROTAVIRUS
 Causing severe diarrheal disease in

humans (30-50% of all cases)
 80 nm in diameter
 Appearance of little wheels viewed
by EM (rota)
 dsRNA in a capsid
 Symptoms: vomiting, watery
diarrhea, fever
 Person with disease can excrete
10^8-10^10 units/ml of feces
 Infective dose 10-100 units
ADENOVIRUSES:
~80 NM DIAMETER
• Only human enteric virus
with Double-stranded DNA
• Protein coat with
attachment fibers
• Infection in lungs, eyes,
genitals
• Possible agent in
waterborne outbreaks
(found in water)
PARAMETERS
 Cryptosporidium parvum - Protozoa
→ Lowest form of animal life
→ Unicellular (complex functional activity)
* Symptoms: Gastrointestinal disorders
* Example – Giardia Lamblia, Entamoeba histolytica
 Exist as a Oocyst (3-6 µm) or Sporozoite
 Infectious dose 1-10 – no effective treatment

GIARDIA LAMBLIA-SINGLE CELLED, PROTOZOA
 Giardiasis - symptoms of mild to

severe diarrhea, nausea and
indigestion
 Exist as a Cyst or Trophozoite.
 The cyst is the environmentally

resistant form ( in water)
 Size: µm 6-8 W × 8-14 L
 Infectious dose < 20

PARAMETERS
 Helminths
→ Parasitic worms
→ Contaminated by aquatic species (snails, insects)
→ Easy to destruct by disinfection
❑ Water borne pathogens → Introduced through fecal

contamination
❑ Intestinal tract organism are good indicator organism
❑ Fecal Coliform group (FC) → for wastewater quality
❑ Total Coliform group (TC) → for water quality

SERIAL DILUTION
IDENTIFICATION OF BACTERIA
 Direct count
 Microscopically or Electronically (based on EC)
→ Differentiation of live and dead cells not possible
→ Stain can be to count different bacteria
 Pour plate method
→ Dilute sample serially
→ Diluted sample + agar medium →then solidify
→ Incubate and count the cells as cfu/mL
 Spread plate method
→ Spread sample on top of the solid medium

IDENTIFICATION OF BACTERIA
 Membrane filtration technique
 Filter sample using 0.45 µm
 Remove the filter paper with solids (microbes)
 Incubate and count the colonies
TESTING FOR COLIFORM BACTERIA:
MEMBRANE FILTRATION
 Membrane filter
 0.45 μm pores
 47 mm in diameter
 Filter 100 mL of water to

be tested through the
membrane filter
MEMBRANE FILTRATION
Add 2 mL of m- Place membrane

Petri dish with endo broth filter in the petri
sterile absorbent (selective dish on top of
nutrient pad media) the nutrient pad
MEMBRANE FILTRATION:
INCUBATION AND RESULTS
 Incubate for 24 hours at
35°C 2
1
 Coliform bacteria grow
4
into colonies with a 3
green metallic sheen 5 6
 Non-coliform bacteria
8
may grow into red 7
colonies
 Coliform concentration is
__________________
8 coliform/100 mL
MULTIPLE TUBE FERMENTATION
TECHNIQUE
 Coliform bacteria → Ferment lactose
 Test has three stages
→ 1. Presumptive test
→ 2. Confirmed test
→ 3. Completed test
 Gas formation is considered as the +ve indication
 From the test results →estimate the MPN
 MPN → Most probable number
 MPN determined by: (1) Poisson distribution, (2)
MPN Tables and (3) Thomas equation
MULTIPLE TUBE FERMENTATION
TECHNIQUE
3
MOST PROBABLE NUMBER
 Poisson distribution
Where,
y → Probability of occurrence of a given result
a → constant for a given set of conditions
n1, n2, n3 → sample size in each dilution
λ → Coliform density, number/mL
p1, p2, p3 → number of positive tubes in each sample dilution
q1, q2, q3 → number of negative tubes in each sample dilution
 Thomas Equation
MOST PROBABLE NUMBER
 The result of a coliform analysis using the
multiple-tube fermentation test of the effluent
from an intermittent sand filter are given below.
Using these data, determine the coliform density
(MPN/100 mL) using the Poisson equation, the
Thomas equation and the MPN Table.
Size of portion, Number Number

mL positive negative
1.0 4 1
0.1 3 2
0.01 2 3
0.001 0 5
λ Ya (x10-7)
SOLUTION 3.80
3.84
3.6754
3.6773
 Poisson Distribution → 3.85 3.6774

3.86 3.6773
MPN/100 mL = 100 x λ
3.90 3.6755
= 385/100 mL
 Thomas Equation
No. of positive tubes =9

mL of sample in –ve tubes = 1.235 mL
mL of sample in all tubes = 5.555 mL
→ MPN/100 mL = 344/100 mL
 MPN Table
→ 4-3-2 ➔ 39 x 10 = 390/100 mL

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CE5170 – PHYSICOCHEMICAL PROCESSES FOR

WATER AND WASTEWATER TREATMENT

Biological Water Quality

→ Capable of infecting or transmitting diseases

 Various disease with symptoms of fever, vomiting, jaundice.

 Size of viruses between 0.02-0.08 µm

 Conc. of viruses in raw wastewater 103-104/100 ml, with infectious

 Found in wastewater and water

 Causing severe diarrheal disease in

 Infectious dose 1-10 – no effective treatment

 Giardiasis - symptoms of mild to

 The cyst is the environmentally

 Infectious dose < 20

❑ Water borne pathogens → Introduced through fecal

❑ Fecal Coliform group (FC) → for wastewater quality

❑ Total Coliform group (TC) → for water quality

→ Spread sample on top of the solid medium

 Filter 100 mL of water to

Add 2 mL of m- Place membrane

Size of portion, Number Number

 Poisson Distribution → 3.85 3.6774

No. of positive tubes =9

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