LABORATORY MANUAL Microbiology, Parasit（簡體書）

Thislaboratorytextisusedtoguidethestudents'learningandtrainingofmicrobiology，parasitologyandimmunologylabtechniques，proceduresandexperiments.Theoutcomeexpectedisstudents'capacityofdoingresearchinlaboratoryofmicrobiology，parasitologyandimmunology.
Themanualconsistsoftwoparts：PartⅠ：outlinesthe8basicexperiments.PartⅡ：outlinesthe21comprehensiveexperimentsofmicrobiology，parasitologyandimmunology.Theinvolvedexperimentsaredesignedtohelpenhanceanddeepenthestudents'un-derstandingofmicrobiology，parasitologyandimmunology.．

《醫學教育改革系列教材:病原生物學與免疫學實驗》由高等教育出版社出版。

Part1BasicExperiments
Experiment1ObservationofBacterialMorphology
Ⅰ.UsingOilImmersionLensoftheMicroscope
Ⅱ.BasicMorphologyandSpecialStructurcofBacteria
Experiment2BacterialStainingandMicroscopy
Ⅰ.GramStain
Ⅱ.Acid-fastStain（Ziehl-NeelsenMethod）
Experiment3PreparationoftheBasicMedium
Experiment4ExaminationMethodsofParasiticInfections
Ⅰ.DirectFecalSmearMethodforDetectingAscarisEggs
Ⅱ.ExaminationforAscarisI.arvaeinLungTissueofMice
Ⅲ.SaturatedBrineFlotationMethodforDetectingHookwormEggs
Ⅳ.FecalCultureMethodforHookwormLarvac
Ⅴ.CellophaneTapeMethodforDetectingPinwormEggs
Ⅵ.DirectSputumSmearorCentrifugalSedimentationMethodforDetectingParagonimusEggs
Ⅶ.SquashMethodforDetectingStriatedMuscleLarvaeofTrichmella
Ⅷ.TestofConcentrationofEggsbySedimentationforDetectingS.IaponicumEggs
Ⅸ.TheMiracidiumHatchingMethodforDetectingS.Japonicum
Ⅹ.CircumovalPrecipitationTest（COPT）forDiagnosingSc.histosomiasis
Ⅺ.DirectStoolSmearwithIodine-stainforDetectingAmoebaCysts
Ⅻ.BloodSmearMethodforDetectingMalarialParasites
Experiment5AgglutinationReaction
Ⅰ.DircctAgglutinationReaction-IdentificationoftheABOBloodGroupSystem
Ⅱ.IndirectAgglutinationReaction：-Anti-streptolysinOTest
Ⅲ.IndirectAgglutinationInhibitionReactionLatexPregnancyTest
Experiment6PrecipitationReaction
Ⅰ.SingleRadialImmunodiffusionTest
Ⅱ.DoublelmmunodiffusionTest
Experiment7Enzyme-IinkedImmunosorbentAssay（SandwichELISAAssay）

Part2ComprehensiveExperiments
Unit1IsolationofMicroorganismsfromtheEnvironmentandHumanBody
Experiment8BacteriaintheAirandUltravioletRadiation
Experiment9BacteriaontheFingerTipandSolidSurface
Unit2DetectionofSkinInfections
Experiment10CulturalCharacteristicsofBacteriaandFungi（TeachingDemonstration）
Ⅰ.ClostridiumPerfringensintheMilkMedium.
Ⅱ.FungiinSabouraudDextroseMedium
Experiment11ObservationofPlasmodium,ToxoplasmaGondii,TrichinellaSpiralis,EchinococcusGranulosusandInsecta
Ⅰ.Plasmodium
Ⅱ.ToxoplasmaGondii
Ⅲ.TrichinellaSpiralis
Ⅳ.EchinococcusGranulosus
Ⅴ.Insecta
Unit3DetectionofRespiratoryInfections
Experiment12Isolation&IdentificationofPyogenicCocci
Ⅰ.IsolationandCultureofMimicPusSwabandThroatSwab
Ⅱ.CoagulaseTest
Experiment13CulturalCharacteristicsofRespiratoryInfectedBacteria
Ⅰ.TheColoniesofPathogenic（JoccionBloodAgarPlates（TeachingDemonstration）
Ⅱ.MycobacteriumTuberculosisinLowensteinJensenSolidMedium（TeachingDemonstration）
Experiment14ObservationofParagonimusWestermani
Unit4DetectionofEntericInfections
Experiment15IsolationandIdentificationofPathogenicEnterobacteria
Ⅰ.IsolationofArtificialFecalSpecimens
Ⅱ.BiochemicalActivitiesofEntericPathogens-KliglerIronAgarTest
Ⅲ.SerologicIdentificationofEntericPathogens
Experiment16WidalTest
Experiment17ObservationofAscarislumbricoides,Hookworms,EnterobiusVermicularis,Taenia,EntamoebaandCryptosporidium
Ⅰ.AscarisLumbricoides
Ⅱ.Hookworms（AncylostomaDuodenaleandNecatorAmericanus）
Ⅲ.EnterobiusVermicularis
Ⅳ.IntestinalCestodes（TaeniaSoliumandTaeniaSaginata）
Ⅴ.IntestinalProtozoa（EntamoebaHistolyticaandEntamoebaColi）
Ⅵ.Cryptosporidium
Unit5DetectionofHepaticInfections
Experiment18ObservationofClonorchisSinensis
Experiment19ObservationofSchistosomaJaponicum
Unit6DetectionofGenitourinaryTractInfections
Experiment20ObservationofChlamydiaTrachomatisInclusion
Unit7AntimicrobiaIResistance
Experiment21BacterialConjugation:theTransferofPlasmidsCodingforAntibioticResistance
……

Appendix
Index．

Ⅱ. Examination for Ascaris Larvae in Lung Tissue of Mice
? Materials
2 % formalin, filter paper, scissors, incubator, glass slides, microscope.
? Procedures
（1） Cultivation of infective eggs
Cover the culture dish with one layer of filter paper moistened with 2%formalin. Dissect female Ascaris, take out about 1.5 cm of one uterus nearthe vagina with forceps and then put all fertilized ova onto the culture dishevenly. Incubate the culture dish in incubator at 25℃ for 3 weeks and keep itmoisture.
（2） Mouse inoculation
Three weeks post cultivation, the ova are mature. Let the mouse starvedfor one day before inoculation. After that, feed the mouse with the incubatedova mixed with food. The mouse will be infected.
（3） Detecting lung tissue of infected mouse
From seven to twelve days after inoculation, sacrifice and dissect the in-fected mouse and take out the lungs. Cut a piece of lung tissue with scissorsand press it between two glass slides and observe the live Ascaris larvae under microscope.
Ⅲ. Saturated Brine Flotation Method for Detecting
Hookworm EggsHookworm eggs can float on the surface of saturated saline solution, as thegravity of saturated saline solution is bigger than that of the eggs. There-fore, this method is the best one for detecting hookworm eggs. And also,eggs from other nematodes, such as Ascaris and Trichuris can be detectedby this method.
? Materials
Fecal sample, saturated salt solution（specific gravity 1.2）,a small peni-cillin vessel, sticks, glass slid, cover slip, microscope.
? Procedures
（1） Add a little amount of saturated salt solution into a small penicillin vessel.
（2） Pick appropriate quantity of fecal sample（the size as one pea）with astick into the penicillin vessel containing saturated salt solution, and mix them thoroughly.
（3） Add saturated salt solution to the penicillin vessel until it is full. Place a glass slide to cover the brim of the vessel.
（4） Leave the vessel for 15- 20 minutes,lift off the glass slide and re-verse it quickly,then place a cover slip on it, and examine under the micro-scope.