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Standard Operating Procedures of Stool Analysis (Stool SOP)

Apr 05, 2015

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[email protected] Standard OperatingProceduresOf Stool [email protected] : -What to LOOK FOR in STOOL :- 1 - : - : a) ( b) ( )protozoa(c) ( .2 : stool culture3 : . : COLLETION OF STOOl . : . - . . : Safety:-- ) ( [email protected] .- .- . 2aI.z 2Lax. : . . . Macroscopic Examination 2aI.z 2Lax. : . . : : 1-consistency-hard and dry-firm and formed-soft and formed-soft and unformed-semi liquid-liquid and watery. 2 : - 2- color :-(a) black .(b) Red ) ([email protected](c) Bright red .(d) Mucous or Clay colored .(e) White .(f) Pale .(g) Green Diathiazine , Mercurous .3 . -3 odor4 . 4- mucus .5 . 5 -Blood : Microscopic examination- : : Microscopic Slides - Cover Slides . Waxes pen or Markers for Labelling . Saline solutions Iodide solution-Loop or applicator sticks : 1 .2 [email protected] ) ( . amoebic trophozoites BMB )(Buffered Methylene Blue ) 5 10 30 . : - X 10 X 40 X 40 . , X 10 . [email protected] : - . ) Protozoa cysts ( , . 2aaL 2Lax. : . . : 1 1gm Iodine2- 2gm Potasium Iodide3-100 ml Di se l ed Wa t er + 30 ) 5 L ( . . Occult Blood- Occul t Bl ood t est ,ant i coagul ant s,ant i met abol i t es,col chi ci nes,i ndomt haci ne,(NSAIDs) Tur ni p Radi shes M el ons 3 . .hayat abdul hami [email protected] 3 Ki t . :- 1 .- 2 AmebaeTr ophozoi t es- 3 Amebae -cyst sTable 1Characteristics of Intestinal Amebae Visible in Different Types of FecalPreparationsUNSTAINED TEMPORARYSTAINSPERMANENTSTAINSCharacteristicSaline FormalinIodine(Cysts)BufferedMethyleneBlue(Trophs)1TrophozoitesMotility +CytoplasmAppearance + + + +Inclusions (rbc., bacteria) + + + +Nucleus +2+ +CystsNuclei + + +Chromatoid bodies + + +3+Glycogen +(vacuole present)hayat abdul hami [email protected] 2Differential Morphology of Protozoa Found in Stool Specimensof Humans: Amebae-TrophozoitesNUCLEUS CYTOPLASMSpeciesSize(Length)MotilityNumberPeripheralChromatinKaryosomalChromatinAppearanceInclusionsEntamoebahistolytica10-60m. Usualrange,15-20mcommensalform.1Over20 minvasiveform.2Progressivewithhyaline,finger-likepseudopods.1Notvisibleinunstainedpreparations.Finegranules. Usuallyevenlydistributed anduniformin size.Small,discrete. Usuallycentrally located,but occasionallyis eccentric.Finelyqranular. Red bloodcellsoccasionally. Noninvasiveorganismsmaycontainbacteria.Entamoebahartmanni5-12m.Usualrange,8-10m.Usuallynonprogressive butmay beprogressiveoccasionally.1Notvisibleinunstainedpreparations.SimilartoE.histolytica.Small, discrete,often eccentric. Finelygranular. Bacteria.Entamoeba coli 15-50m.Usualrange,20-25m.Sluggish,nonprogressive, withbluntpseudopods.1Oftenvisibleinunstainedpreparations.Coarsegranules,irregular in sizeanddistribution.Large, discrete,usually eccentric. Coarse,oftenvacuolated.Bacteria,yeasts,othermaterials.hayat abdul hami [email protected],15-20m.Usuallysluggish,similarto E.coli. Occasionally, indiarrheicspecimens,motilitymay beprogressive.1May beslightlyvisibleinunstainedpreparations.Occasionallymaybeirregularlydistorted bypressurefromvacuoles incytoplasm.Usuallyfinegranulesevenlydistributed. Occasionallygranules maybeirregularlyarranged. Chromatinsometimes inplaquesorcrescents.Small, discrete,eccentric. Occasionally large,diffuse orirregular.Coarsely,granular,mayresembleE.coli. Containsnumerousvacuoles.Bacteria,yeasts.Endolimax nana 6-12m. Usualrange,8-10m.Sluggish,usuallynonprogressive withbluntpseudopods.1Visibleoccasionally inunstainedpreparations.None. Large, irregularlyshaped, blot-like. Granular,vacuolated.Bacteria.Iodamoebabuetschlii8-20m. Usualrange,12-15m.Sluggish,usuallynonprogressive.1Notusuallyvisibleinunstainedpreparations.None. Large, usuallycentral. Surrounded by refractile,achromaticgranules. Thesegranules areoften not distincteven in stainedslides.Coarselygranular,vacuolated.Bacteria,yeasts, orothermaterial.hayat abdul hami [email protected] Usualrange,9-12m.Pseudopodsareangular,serrated, orbroadlobed,andhyaline,almosttransparent.2(Inapproximately 20%oforganismsonly 1nucleus ispresent.) Nucleiinvisible inunstainedpreparations.None. Large cluster of4-8 granules. Finely,qranular. Bacteria:occasionally redblood cells.hayat abdul hami [email protected] Morphology of Protozoa Found in Stool Specimens of Humans:Amebae-cystCYTOPLASM NUCLEUSGlycogen ChromatoidBodiesKaryosomalChromatinPeripheralChromatinNumber Shape Size(Diameter orlength)SpeciesUsuallydiffuse. Concentratedmass oftenpresent inyoungcysts. Stains reddishbrown withiodine.Present.Elongatedbars withbluntlyroundedends.Small,discrete,usuallycentrallylocatedPeripheralchromatinpresent.Fine,uniformgranules, evenlydistributed4 inmaturecyst.Immature cystswith 1 or2occasionally seenUsuallyspherical.10-20mUsualrange,12-15mEntamoebahistolyticaSimilar toE.histolytica.Present.Elongatedbars withbluntlyroundedends.Similar toE.histolytica.Similarto E.histolytica.4 inmaturecyst.Immature cystswith 1 or2 oftenseenUsuallyspherical.5-10mUsualrange,6-8 m.EntamoebahartmanniUsuallydiffuse,but,occasionally welldefinedmass inimmaturecysts. Stain reddishbrown withiodine.Present,but lessfrequentlyseen thanin E.histolytica. Usuallysplinter-like withpointedends.Large,discrete,usuallyeccentricbutoccasionally centrallylocated.ripheralchromatinpresent.Coarsegranulesirregularin sizeanddistribution, butoftenappearmoreuniformthan introphozoites.8 inmaturecyst. Occasionally super-nucleated cystswith 16or moreare seen.Immature cystswith 2 ormoreoccasionally seen.Usuallyspherical. Occasionallyoval,triangular, orothershapes.10-35mUsualrange,15-25mEntamoeba coliUsuallysmall,diffusemassesstainreddishbrown withiodine. Adark areacalled an"inclusionmass"(possiblyPresent.Manysmallbodieswithangularorpointedends, orfew largeones. May be oval,rod-like,Usuallysmall andeccentric.Usuallyfinegranulesevenlydistributed1. Rarely2.Occasionallyvisible inunstainedpreparationsSpherical or oval.-18 mUsualrange,11-15m.Entamoebapoleckihayat abdul hami [email protected])is oftenalsopresent. Mass doesnot stainwithiodine.orirregular.Usuallydiffuse. Concentratedmass seenoccasionally in youngcysts. Stains reddishbrown withiodineOccasionallygranulesor smallovalmassesseen, butbodies asseen inEntamoeba spp. arenotpresent.Large(blot-like),usuallycentral.None 4 inimmature cysts.Immature cystswith lessthan 4rarelyseen.Spherical,ovoidal,orellipsoidal.5-10m.Usualrange,6-8 m.EndolimaxnanaCompact,well-definedmass. Stains darkbrown withiodine.Occasionallygranulespresent,butchromatoid bodiesas seen inEntamoeba spp. arenotpresent.Large,usuallyeccentric.Refractile,achromaticgranuleson one sideofkaryosome.Indistinctin iodinepreparationsNone 1 inmaturecyst.Ovoidal,ellipsoidal,triangular, orothershapes5-20m.Usualrange,10-12mIodamoebabuetschliihayat abdul hami [email protected] Nor mal Values in St ool AnalysisNor mal Val ues M acr oscopi c Exami nat i on100-200g/ 24h AmountBr own Col orOf f ensi ve- var i es wi t h PH of st ool anddepends on bact er i al f er ment at i on andput r ef act i onOdorPl ast i hc ,not unusual t o seef i ber ,veget abl eski ns,and seeds,sof t andbul ky i n hi gh veget able diet ,smal l anddr y i n hi gh meat dietConsi st encyFor med Si ze and shapeNone Gr oss bl oodNone M ucusNone PusNone Par asi t esNor mal Val ues M i cr oscopi c Exami nat i onCol or l ess ,neut r al f at (18%)and f ay aci dcr yst al s and soapsFatNone t o smal l amount Undi gest ed f ood ,meat f iber s ,t r ypsin,st ar chNone Eggs and segment s of par asi t esNone Bact er i a and vi rusesNone Yaest sNone Leukocyt esNor mal Val ues Cl i ni cal Exami nat i onUp t o 75% Wat erNeut r al t o weakl y al kal i ne (PH 7.0 7.5) PHNegat i ve Occul t bl ood50 300gm/ 24h Uor obi l inogenCor popor phyr i ns 400-1200ug/ 24hUr oporphyr i ns10-40ug/ 24hPor phyr i ns2.5g/ 24h Ni t r ogen20 -95U/ g Tr ypsi n5.8- 9.8mmol / d Sodi um2.5 3.9 mmol / d Chl or i de15.7 20.7 mmol/ d Pot assi um0 - -6gm / 24h Li pi ds (f at t y aci ds )2.5 g/ L Car bohydrat es (as r educing subst ances)hayat abdul hami [email protected] : -Protozoa Found in Stool Specimens of Humans: AmebaeAmebaeEntamoebahistolytica/dispar Entamoebahartmanni Entamoebacoli Entamoebapolecki Endolimaxnana Iodamoebabeutschliihayat abdul hami [email protected] Found in Stool Specimens of Humans:Ciliate and FlagellatesCiliate FlagellatesBalantidiumcoli Chilomastixmesnili Giardiaintestinalis Dientamoebafragilisnocyst stage_______40 m______10 mhayat abdul hami [email protected] Found in Stool Specimens of Humans:Coccidia and BlastocystisCoccidia BlastocystisCystoisosporabelli Sarcocystisspp. Cryptosporidiumspp. Cyclosporacayetanensis Blastocystishominiswet mount oocyst wet mount wet mount wet mountsafranin sporocyst acid-fast acid-fast trichrome_____10 mhayat abdul hami [email protected] Found in Stool Specimens of Humans Trematode E Clonorchissinensis Opisthorchisviverrini Paragonimuswesterm