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    Examination of Specimens For Parasites

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    Jhost Clinton Purba
    This document provides guidance on examining specimens for parasites. It discusses the basic laboratory equipment and facilities needed, as well as methods for examining different specimen types like feces, blood, biopsy materials, and cultures. Specific techniques are described for concentrating and examining specimens for parasites, including various staining and transport methods. The goal is to reliably detect, identify, and in some cases quantify different parasite stages and species.

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    Examination of Specimens For Parasites

    Uploaded by

    Jhost Clinton Purba
    82 views32 pages
    This document provides guidance on examining specimens for parasites. It discusses the basic laboratory equipment and facilities needed, as well as methods for examining different specimen types like feces, blood, biopsy materials, and cultures. Specific techniques are described for concentrating and examining specimens for parasites, including various staining and transport methods. The goal is to reliably detect, identify, and in some cases quantify different parasite stages and species.

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    Examination of Specimens for Parasites

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    This document provides guidance on examining specimens for parasites. It discusses the basic laboratory equipment and facilities needed, as well as methods for examining different specimen types like feces, blood, biopsy materials, and cultures. Specific techniques are described for concentrating and examining specimens for parasites, including various staining and transport methods. The goal is to reliably detect, identify, and in some cases quantify different parasite stages and species.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    82 views32 pages

    Examination of Specimens For Parasites

    Uploaded by

    Jhost Clinton Purba
    This document provides guidance on examining specimens for parasites. It discusses the basic laboratory equipment and facilities needed, as well as methods for examining different specimen types like feces, blood, biopsy materials, and cultures. Specific techniques are described for concentrating and examining specimens for parasites, including various staining and transport methods. The goal is to reliably detect, identify, and in some cases quantify different parasite stages and species.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    of 32

    EXAMINATION OF SPECIMENS FOR PARASITES

    dr. RONALD TAMBUNAN, MKT

    PREFACE
    LAB DIAGNOSIS IS A BASIC STEP IN THE

    EVALUATION OF DISEASE PROCESSES (CONFIRMING A PRESUMPTIVE CLINICAL DIAGNOSIS & PROVIDING EVIDENCE OF AN UNSUSPECTED AGENT OF DISEASE) EXAMINATION OF SPECIMENS INVOLVES 2 STEPS: 1. DETECTION OF PARASITE (SKILLS OF CARRYING OUT PROCEDURE) 2. IDENTIFICATION (SKILLS IN RECOGNIZING) ALSO NEEDS EXPERIENCES TO DISTINGUISH PARASITE FROM ARTIFACTS (FALSE POSITIVE/NEGATIVE) RELIABLE PARASITOLOGY SERVICE: NEED APPROPRIATE AND ADEQUATE FACILITIES,

    PREFACE (contd.)
    THE MOST BASIC ITEM:

    1.

    2. 3. 4.

    5.
    6.

    A MICROSCOPE EQUIPPED WITH A SUITABLE LIGHT SOURCE & CLEAR, CLEAN LENSES AN OCULAR MICROMETER & A CALIBRATING MEANS A CENTRIFUGE AN INCUBATOR A DRYING OVEN A MEANS OF STERILIZING GLASSWARE & INSTRUMENTS

    PREFACE (contd.)
    SIZE IS AN IMPORTANT CHARACTERISTIC OF

    ALL LIVING THINGS MICROSCOPE MAGNIFIES THE SIZE OF OBJECTS & WITH THE SEVERAL LENS COMBINATIONS, OBJECTS ARE SEEN AT SEVERAL DIFFERENT MAGNIFICATIONS FOR RECOGNIZING & DESCRIPTING PARASITES & THEIR VARIOUS STAGES, A MICROSCOPE MUST BE PROVIDED WITH A CALIBRATED MICROMETER SCALE

    EXAMINATION OF FECES
    A FECAL EXAMINATION INVOLVES RECOGNIZING

    & RECORDING : 1. GROSS ELEMENTS 2. DISTINCTIVE PATHOLOGICAL ODORS OR COLORS 3. UNUSUAL QUANTITIES OF NORMAL OR ABNORMAL ELEMENTS (BLOOD, CELLULAR EXUDATE, TISSUE FRAGMENTS, CHARCOTLEYDEN CRYSTALS, & UNDIGESTED FOOD) 4. RESIDUES OF MEDICATIONS (MINERAL OIL, BISMUTH, IRON, MAGNESIUM, OR BARIUM) 5. DETECTING THE PRESENCE OR ABSENCE OF PARASITES

    EXAMINATION OF FECES (contd.)


    STOOL SPECIMENS USUALLY ARE

    DESCRIBED WITH REFERENCE TO 3 QUALITIES: 1. CONSISTENCY 2. COLOR 3. ELEMENTS AN ODOR WOULD ASSIST IN INTERPRETING THE OTHER FINDINGS A TERM USED IN DESCRIBING AN ODOR IS FOUL-SMELLING (SUBJECTIVE)

    STOOL CLASSIFICATION & CODE


    CONSISTENCY 1. HARD, RESIST PUNCTURE 2. FORMED, CAN BE PUNCTURED 3. SOFT, CAN BE CUT WITH APPLICATOR 4. MUSHY, CAN BE RESHAPED 5. LOOSE, SHAPE TO CONTAINER 6. DIARRHEIC, FLOWS 7. WATERY, POURS 1. 2. 3. 4. 5. 6. 7. 8. COLOR BLACK DARK BROWN BROWN PALE BROWN YELLOW GREEN CLAY OTHER* ELEMENTS 1. PULP & FIBER NEARLY PURE 2. CONSPICUOUSLY FIBROUS 3. FIBER MODERATE TO SCANTY 4. PURE COLLOIDAL FECES 5. FECES WITH SCANTY MUCUS 6. FECES WITH MUCH MUCUS 7. MUCUS WITH SCANTY FECES+ 8. OTHER (BLOOD, BARIUM)

    STOOL CLASSIFICATION & CODE (contd.)


    *COLOR, SUCH AS BLUE, RED, OR PURPLE, CAN INDICATE SPECIAL MEDICATION +A SPECIMEN CONTAINING GROSS BLOOD & MUCUS, WITH OR WITHOUT PUS, WOULD BE RECORDED AS DYSENTRIC & WOULDNT BE CODED FOR CONSISTENCY OR COLOR

    COLLECTION OF THE SPECIMEN


    COLLECTED IN A CLEAN CONTAINER

    WITHOUT CONTAMINATION WITH URINE, WATER, etc. AMOUNT: WHOLE STOOL, SERIES OF STOOLS OVER A SPECIFIED PERIOD, MILIGRAMS SCRAPED FROM MEDICAL GLOVE USED AFTER RECTAL EXAMINATION ROUTINE: 1. 2 5 GRAM, PROTECTED FROM DRYING 2. FREE OF OIL, BARIUM & BISMUTH 3. BETTER TAKEN FROM NORMALLY PASSED STOOL THAN PURGATION OR ENEMA

    COLLECTION OF THE SPECIMEN (contd.)


    A FORMED STOOL CAN BE KEPT OVERNIGHT

    AT MODERATE OR LOW ROOM TEMPERATURE DIARRHEIC & DYSENTRIC STOOLS SHOULD BE EXAMINED PROMPTLY OR PRESERVED OTHERS SHOULD BE EXAMINED IN 3 4 HRS PRESERVATION METHODS: 1. REFRIGERATOR 2. CHEMICALLY (FORMALIN, POLYVINYL ALCOHOL (PVA), MERTHIOLATE-IODINEFORMALIN (MIF), SCHAUDINNS FIXATIVE)

    CHOICE OF METHOD
    DIRECT EXAMINATION

    DIRECT SALINE SMEAR 2. KATO THICK SMEAR PERMANENT, STAINED, FRESH FECAL SMEAR 1. IRON-HEMATOXYLIN-STAINED SMEAR 2. TRICHROME-STAINED SMEAR 3. PVA SMEAR 4. SCHAUDINNS PRESERVED SPECIMEN 5. MIF PRESERVED SPECIMEN
    1.

    CONCENTRATION METHODS FOR FECAL SPECIMENS


    GOAL: SEPARATE CYSTS OF PROTOZA &

    EGGS OF HELMINTHS FROM OTHER ELEMENTS OF FECES HOW TO ACHIEVE: SEDIMENTATION, FLOTATION, OR COMBINING OF THE 2 SEDIMENTATION: SUSPENDING THE SPECIMEN IN WATER OR AN AQUEOUS SOLUTION OR BY CENTRIFUGATION FLOTATION : SUSPENDING THE SPECIMEN IN A MEDIUM OF GREATER DENSITY THAN CYSTS & EGGS

    CONCENTRATION METHODS FOR FECAL SPECIMENS (contd.)


    SEDIMENTATION IN WATER:

    THE RESULT DEPENDS ON THE ACCURACY OF TIMING OF SEDIMENTATION 2. SKILLS 3. EFFICACY OF THE STRAINING PROCEDURES THUS IT ISNT GENERALLY USED MODIFICATION METHOD FOR CONCENTRATING: USE OF ETHER (ABSORBED BY FECAL DEBRIS AND CONSEQUENTLY BECOMES LIGHTER THAN WATER) ETHER: FLAMMABLE & EXPLOSIVE
    1.

    CONCENTRATION METHODS FOR FECAL SPECIMENS (contd.)


    FORMALIN-ETHER SEDIMENTATION

    ACID-ETHER SEDIMENTATION
    FLOTATION METHODS

    BRINE FLOTATION 2. ZINC SULFATE CENTRIFUGAL FLOTATION


    1.

    SCHISTOSOME EGGHATCHING
    DETECTING EGGS IN FECES

    DETECTING MIRACIDIA IN FECED, URINE,

    MACERATED TISSUES, OR POST MORTEM INTESTINAL SCRAPPINGS CONCENTRATING TECHNIQUE: USING A SIDEARM FLASK OR ERLENMEYER FLASK & TEST TUBE WITH FILTER

    QUANTITATIVE DIAGNOSIS
    TECHNIQUES FOR BOTH QUANTITATIVE &

    QUALITATIVE: DIRECT SALINE & THE CELOPHANE-COVERED THICK SMEAR OTHER TECHNIQUE: UNRELIABLE FOR EGG COUNTS BECAUSE THE YIELDS ARE VARIABLE, DUE TO DIFFERENCES IN THE CHARACTER OF SPECIMENS & THE PROFICIENCY OF TECHNICIANS QUANTITATIVE TECHNIQUE: 1. DIRECT-SMEAR EGG COUNTS 2. DILUTION EGG COUNTS 3. THICK-SMEAR EGG COUNTS 4. DILUTION-FILTRATION EGG COUNTS

    FECAL CULTURE FOR DIAGNOSIS OF NEMATODE LARVAE


    TEST-TUBE CULTURES (HARADA-MORI

    CULTURES) CHARCOAL CULTURES FILTER PAPER SLANT CULTURE (LITTLE, 1966) WORMS MIGRATING FORM THE ANUS: PRESERVED IN ALCOHOL (70%) OR FORMALIN (5 10%) ANAL SCRAPINGS & SWABS: E. HISTOLYTICA & ENTEROBIUS RECOVERY OF WORMS FROM STOOLS: 1. STRAINING AN AQUEOUS SUSPENSION OF THE SPECIMEN THROUGH WIRE SIEVES

    OTHER SPECIMENS
    URINE 1. TRICHOMONAS VAGINALIS 2. SCHISTOSOMA HAEMATOBIUM 3. STRONGYLOIDES (RARE) SPUTUM & GASTRIC WASHINGS 1. TROPHOZOITES OF E. HISTOLYTICA 2. ECHINOCOCCUS GRANULOSUS 3. EGGS OF P. WESTERMANI 4. EGGS OF STRONGYLOIDES 5. ASCARIS 6. NECATOR 7. ANCYLOSTOMA

    OTHER SPECIMENS (contd.)


    BLOOD

    1.
    2. 3. 4. 5.

    MALARIA BABESIOSIS AFRICAN TRYPANOSOMIASIS FILARIASIS CHAGAS DISEASE

    PROTOZOA IN BLOOD
    THIN FILM

    THICK FILM
    MICROFILARIAE

    HEMATOXYLIN-STAINED BLOOD FILM 2. CONCENTRATION OF MICROFILARIAE (KNOTTS METHOD) 3. MEMBRANE FILTRATION


    1.

    EXAMINATION OF ASPIRATE
    PROCTOSCOPIC ASPIRATE: NONMOTILE

    TROPHOZOITE OR CYSTS OF E. HISTOLYTICA DUODENAL ASPIRATION: G. LAMBLIA, S. STERCORALIS, FASCIOLOPSIS BUSKI ASPIRATION FROM LIVER & LUNG LESIONS ASPIRATES FROM LYMPH NODES, SPLEEN, LIVER, BONE MARROW, & SPINAL FLUID: AFRICAN TRYPANOSOMIASIS, VISCERAL LEISHMANIASIS (KALA-AZAR), CHAGAS DISEASE & TOXOPLASMOSIS

    BIOPSY MATERIAL
    SKIN

    AMEBIASIS CUTIS 2. CUTANEOUS LEISHMANIASIS 3. O. VOLVULUS 4. MANSONELLA STREPTOCERCA & MANSONELLA OZZARDI LYMPH NODE 1. AFRICAN TRYPANOSOMIASIS 2. KALA-AZAR 3. CHAGAS DISEASE 4. TOXOPLASMOSIS
    1.

    BIOPSY MATERIAL (contd.)


    MUSCLE

    TRICHINELLA LARVAE 2. T. SOLIUM CYSTICERCUS 3. ANCYLOSTOMA LARVAE 4. TOXOCARA LARVAE COLON & RECTUM 1. S. MANSONI 2. S. JAPONICUM 3. S. HAEMATOBIUM
    1.

    CULTURE METHOD

    CULTIVATION OF PROTOZOA
    AMEBAE & OTHER INTESTINAL PROTOZOA

    1.
    2. 3. 4. 5.

    BALAMUTHS MONOPHASIC MEDIUM BOECK & DRBOHLAVS DIPHASIC MEDIUM AXENIC CULTIVATION OF E. HISTOLYTICA AXENIC CULTIVATION OF GIARDIA IN VITRO EXCYSTATION OF GIARDIA

    CULTIVATION OF PROTOZOA (contd.)


    GENITOURINARY PROTOZOA: T. VAGINALIS

    TRUSSEL & JOHNSONS MEDIUM 2. SIMPLIFIED TRYPTICASE SERUM (STS) MEDIUM CEREBROSPINAL PROTOZOA: NAEGLERIA & ACANTHAMOEBA 1. CULBERTSONS MEDIUM 2. WILLAERTS MEDIUM
    1.

    CULTIVATION OF PROTOZOA (contd.)


    BLOOD & TISSUE PROTOZOA

    1.
    2. 3. 4. 5. 6.

    NNN (NOVY & MacNEAL) MEDIUM HOCKMEYERS MEDIUM WEINMANS MEDIUM TOBIES DIPHASIC MEDIUM CULTIVATION OF MALARIA PARASITES (BASS & JOHNS) USING RPMI 1640 MEDIUM XENODIAGNOSIS

    SPECIMENS TRANSPORT
    STUARTS TRANSPORT MEDIUM

    AMIES TRASNPORT MEDIUM


    CARY-BLAIR MEDIUM

    STAINING
    GRAM

    ZIEHL-NEELSEN
    HAEMATOXYLIN & EOSIN (H&E) PAPANICOLAOU PERIODIC ACID-SCHIFF (PAS) ROMANOWSKY SILVER SUDAN CONKLINS

    STAINING (contd.)
    ACRIDINE ORANGE BISMARCK BROWN CARMINE COOMASSIE BLUE CRYSTAL VIOLET

    DAPI
    EOSIN ETHIDIUM BROMIDE ACID FUCHSINE HAEMATOXYLIN IODINE

    STAINING (contd.)
    MALACHITE GREEN METHYL GREEN METHYLENE BLUE NEUTRAL RED NILE RED

    OSMIUM TETRAOXIDE
    RHODAMINE SAFRANIN

    PHOSPHOTUNGSTIC ACID
    RUTHENIUM TETROXIDE

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