微生物肠杆菌科培训ppt课件

微生物微生物肠肠杆菌科杆菌科微生物肠杆菌科1Outline GeneralpropertiesofentericbacilliEscherichia（埃希菌属）Shigella（志贺菌属）Salmonella（沙门菌属）微生物肠杆菌科2OutlineGeneralpropertiesof1.MorphologyGram-negativeRodshaped(bacilli)Mostformpiliandperitrichousflagella(周鞭毛)Donotformspores2.Culture characteristicsFacultativeanaerobes(兼性厌氧)oraerobes(需氧)Growreadilyonsimplemedia.Colonies:circular,convex,smoothBloodagarplate:somecanformhemolyticzones.Liquidmedia:turbid.微生物肠杆菌科3Morphology微生物肠杆菌科33.Biochemical reactionsCatalase（触酶）test:+Reducenitrate(硝酸盐)tonitrite(亚硝酸盐)Oxidase(氧化酶)test:-Lactose fermentation test(乳糖发酵试验)乳糖发酵试验可用于初步鉴别肠道致病菌和非致病菌肠道致病菌多数不发酵乳糖（沙门菌、志贺菌）肠道非致病菌多数发酵乳糖（埃希菌）微生物肠杆菌科4Biochemicalreactions微生物肠杆菌科44.AntigensO antigens:LPS,IgMH antigens:flagellin,IgGCapsular antigens:typespecific,ViantigenandKantigen.5.Resistance:relativelyweak6.Variation微生物肠杆菌科5Antigens微生物肠杆菌科5Gram-negative rod-shaped bacilli微生物肠杆菌科6Gram-negativerod-shapedbacilMacconkey agar with lactose(left)and non-lactose(right)fermenters.微生物肠杆菌科8Macconkeyagarwithlactose(le微生物肠杆菌科9微生物肠杆菌科9Section 1 Escherichia(埃希菌属)E.coli微生物肠杆菌科12Section1Escherichia(埃希菌属)E.coliE.coli1.Biologicalcharacterization2.PathogenesisandsyptomatologylPathogenicsubstanceslClinicalfindings3.Laboratorydiagnosis4.Treatment微生物肠杆菌科13E.coliBiologicalcharacteriza1.MorphologyGram-negativeRodshaped(bacilli)Mostformpiliandperitrichousflagella(周鞭毛)Donotformendospores2.Culture characteristicsFacultativeanaerobes(兼性厌氧)Liquidmedia:turbid.Growreadilyonsimplemedia.Colonies:circular,convex,smooth.微生物肠杆菌科14Morphology微生物肠杆菌科143.Biochemical reactionsLactosefermentationtest(乳糖发酵试验):(SSagar,KIagar)H2Stest:-Motilitytest:+IMViCtests:+-4.Antigens:Oantigens:170typesHantigens:50typesKantigens:100types微生物肠杆菌科15Biochemicalreactions微生物肠杆菌科15Gram-negative rod-shaped bacilli微生物肠杆菌科16Gram-negativerod-shapedbacilSS agar(Salmonella Shigella agar)Ingredients:Nutrient agar 100ml Lactose(乳糖乳糖)1.00g Bile salts 0.85g inhibitor Sodium citrate 0.85g inhibitor Sodium thiosulfate 0.85g 10%ferric citrate 1ml 1%Neutral red 0.25ml indicator 1%brilliant green 0.033ml inhibitor微生物肠杆菌科17SSagar(SalmonellaShigellaEscherichia coli on SS agar(pink colony)微生物肠杆菌科18EscherichiacolionSSagar(pKligler Iron Agar(KI agar)Ingredients:Peptones 2.00 g NaCl 0.50g Glucose 0.10g Lactose 1.00 g Ferrous sulfate 0.05g Sodium thiosulfate 0.075g Agar 0.50g 0.2 phenol red 1.25ml Distilled water 100mlSlantButtIn acid condition:yellowIn base condition:red微生物肠杆菌科19KliglerIronAgar(KIagar)IngReading results of KI agar(sulfureted hydrogen)微生物肠杆菌科20ReadingresultsofKIagar(sul2-Shigella 3-Salmonella 4-E.Coli微生物肠杆菌科212-Shigella3-Salmonella4-E.Motility testThe test is based on the flagellum of the bacteria.If the bacteria have flagellum,the semisolid medium will be clouding and the test will be positive.Semisolid medium微生物肠杆菌科22MotilitytestThetestisbaseE.coliSS agar:reddishcolonyKI agar:yellowslantsandbutts,bubbleinthebottom,thelineofinoculationisblurrynoH2SSemisolid:turbidgrowth,withflagella微生物肠杆菌科23E.coliSSagar:reddishcolonyIndole tests色氨酸吲哚对二甲基氨基苯甲醛玫瑰吲哚+E.coli微生物肠杆菌科24Indoletests色氨酸吲哚对二甲基氨基苯甲醛玫瑰吲哚Methyl red tests某些细菌在糖代谢过程中，分解葡萄糖产生丙酮酸，丙酮酸可进一步分解，产生甲酸、乙酸、乳酸等，使培养基的pH降至4.5以下，当加入甲基红试剂则呈红色，为甲基红试验阳性。若细菌分解葡萄糖产酸量少，或产生的酸进一步转化为其他物质（如醇、酮、醚、气体和水等），则培养基的酸度仍在pH6.2以上，故加入甲基红指示剂呈黄色，是为阴性。微生物肠杆菌科25Methylredtests某些细菌在糖代谢过程中，分解Voges-Proskauer test某些细菌在葡萄糖蛋白胨水培养基中能分解葡萄糖产生丙酮酸，丙酮酸缩合，脱羧成乙酰甲基甲醇，后者在强碱环境下，被空气中氧氧化为二乙酰，二乙酰与蛋白胨中的胍基生成红色化合物，称V-P（+）反应。大肠埃希菌V-P试验阴性，产气肠杆菌V-P试验阳性。微生物肠杆菌科26Voges-Proskauertest某些细菌在葡萄糖蛋白Citrate utilization test 以枸橼酸钠为唯一碳源、PH值7.0的培养基上，产气杆菌分解枸橼酸钠产生碳酸盐，使培养基由中性变为碱性，培养基中指示剂溴麝香草酚蓝（BTB）由浅绿色变为深蓝色，此为枸橼酸盐利用试验阳性。大肠杆菌因不能利用枸橼酸盐，此试验为阴性反应。微生物肠杆菌科27Citrateutilizationtest以枸橼酸钠微生物肠杆菌科28微生物肠杆菌科28微生物肠杆菌科29微生物肠杆菌科291.Adhesins2.Exotoxins3.Other virulence factorslEndotoxinlCapsuleslTypesecretionsystemsPathogenic substances微生物肠杆菌科30AdhesinsPathogenicsubstances微Adhesins（黏附素）Function:facilitatethespecificadherence.Several types:lColonizationfactorantigen,CFA/（定植因子抗原,）lAggregativeadherencefimbriae,AAF/（集聚黏附菌毛和）lBundleformingpili,Bfp（束形成菌毛）lIntimin（紧密黏附素）lPpili（P菌毛）微生物肠杆菌科31Adhesins（黏附素）Function:facilitExotoxins（外毒素）Shigatoxins(Stx-1,Stx-2)（致贺毒素和）Heatstableenterotoxins(STa,STb)（耐热肠毒素a和b）Heatlabileenterotoxins(LT-,LT-)（不耐热肠毒素和）Hemolysin（溶血素A）:uropathogenicE.coli(UPEC)微生物肠杆菌科32Exotoxins（外毒素）Shigatoxins(StClinicalfindings1.Extraintestinalinfections（肠道外感染）Septicemia（败血症）Meningitisofnewborn（新生儿脑膜炎）Urinarytractinfection(UTI)（泌尿道感染）-UPEC,urophathogenicE.coli（尿路致病性大肠埃希菌）2.Intestinalinfections（胃肠炎）-Diarrhea-causingE.coli微生物肠杆菌科33ClinicalfindingsExtraintestin微生物肠杆菌科34微生物肠杆菌科34Diarrhea-causing E.coli1.ETEC:enterotoxigenic E.coli肠产毒型大肠埃希菌“Travelersdiarrhea”&ChildrensdiarrheaWaterydiarrheawithoutbloodSelf-limiting(34days)2.EIEC:enteroinvasiveE.coli3.EPEC:enteropathogenicE.coli4.EHEC:enterohemorrhagicE.coli5.EAEC:enteroaggregativeE.coli微生物肠杆菌科35Diarrhea-causingE.coliETEC:Enterotoxigenic(ETEC)Pathogenic substances A.Enterotoxinsa)Heatlabileenterotoxin,LT-b)Heatstableenterotoxin,ST-aB.Colonizationfactorantigens(CFA/)微生物肠杆菌科36Enterotoxigenic(ETEC)PathogenHeat labile enterotoxin（不耐热肠毒素）（不耐热肠毒素）EncodedbyplasmidLT-:amajorvirulencefactorHeatlabileLikecholeragen(霍乱肠毒素)1subunitA(toxic)+5subunitB(binding)SubunitB:bindtoGM1-ganglioside（神经节苷脂）SubunitA:a)Activatesadenylcyclase(腺苷环化酶)b)ATPcAMPc)Secretionwater/ions微生物肠杆菌科37Heatlabileenterotoxin（不耐热肠毒素霍乱肠毒素的作用机制霍乱肠毒素的作用机制微生物肠杆菌科38霍乱肠毒素的作用机制微生物肠杆菌科38Heat stable enterotoxin（耐热肠（耐热肠毒素）毒素）EncodedbyplasmidSTa:amajorvirulencefactorHeatstableFunctions:a)Activatesguanylylcyclase(鸟苷环化酶)b)cGMPc)Secretionwater/ions微生物肠杆菌科39Heatstableenterotoxin（耐热肠毒素）LT-&STaTheresultisthestimulationofchloridesecretionoutofthecellandtheblockageofNaClabsorption.Theneteffectistheaccumulationofwaterandelectrolytesintothebowellumen.微生物肠杆菌科40LT-&STaTheresultisthe环境和食品卫生学上，环境和食品卫生学上，“大肠菌群数大肠菌群数”常被用常被用作粪便污染的检测指标作粪便污染的检测指标 3个大肠菌群个大肠菌群/1L饮水饮水 5个大肠菌群个大肠菌群/100ml瓶装汽水、果汁瓶装汽水、果汁 卫生学指标卫生学指标我国饮用水卫生标准：我国饮用水卫生标准：微生物肠杆菌科41环境和食品卫生学上，“大肠菌群数”常被用作粪便污染的检测指标微生物肠杆菌科42微生物肠杆菌科42微生物肠杆菌科43微生物肠杆菌科43Section 2 Shigella 微生物肠杆菌科44Section2Shigella微生物肠杆菌科44微生物肠杆菌科45微生物肠杆菌科45微生物肠杆菌科46微生物肠杆菌科46Biological characterizationNoflagellaGlucosefermentationtest:+Lactosefermentationtest:-H2Stest:-Motilitytest:-Antigens:OandKantigensS.dysenteriae(GroupA)：痢疾志贺菌S.flexneri(GroupB)：福氏志贺菌福氏志贺菌S.boydii(GroupC)：鲍氏志贺菌S.sonnei(GroupD)：宋内志贺菌宋内志贺菌微生物肠杆菌科47BiologicalcharacterizationNo微生物肠杆菌科48微生物肠杆菌科48Shigella(small,colorless,transparent colony)E.coli(large,reddish colony)微生物肠杆菌科49Shigella(small,colorless,tranShigellasonnei on blood agar.Medium sized,grey colonies that cause no alteration of the blood.微生物肠杆菌科50Shigellasonneionbloodagar.2-Shigella 3-Salmonella 4-E.Coli微生物肠杆菌科512-Shigella3-Salmonella4-E.Motility testThe test is based on the flagellum of the bacteria.If the bacteria have flagellum,the semisolid medium will be clouding and the test will be positive.Semisolid medium微生物肠杆菌科52MotilitytestThetestisbaseShigellaSSagar:flaxencolonyKIagar:redslantsandyellowbutts,nobubbleinthebottom,thelineofinoculationisclearnoH2SSemisolid:lineargrowth,withoutflagella微生物肠杆菌科53ShigellaSSagar:flaxencolonyPathogenesis Pathogenic substances1.Invasiveness2.Endotoxin3.ExotoxinClinical findingsBacillarydysentery(菌痢)HumansareonlyknownreservoirsOral-fecaltransmissionAcuteChronic:2M微生物肠杆菌科54PathogenesisPathogenicsubstInternalized shigellae induce apoptosis of macrophage and release of the bacteriaAttracted by the cytokines released by macrophageActivates the invasion genes on the virulence plasmid M cell3.Exotoxin2.Endotoxin1.Invasiveness微生物肠杆菌科55Internalizedshigellaeinducen靶细胞靶细胞：回肠末端和结肠部位的黏膜上皮细胞：回肠末端和结肠部位的黏膜上皮细胞 首先侵犯派伊尔淋巴结的首先侵犯派伊尔淋巴结的M M细胞细胞n致病过程致病过程：黏附：黏附型分泌系统型分泌系统细胞膜内陷、细菌细胞膜内陷、细菌内吞内吞胞内生长繁殖胞内生长繁殖扩散扩散n脓血黏液便脓血黏液便：坏死的黏膜、死亡的：坏死的黏膜、死亡的WBC、细胞碎片、细胞碎片、纤维蛋白和血液纤维蛋白和血液1 1、侵袭力：、侵袭力：微生物肠杆菌科56靶细胞：回肠末端和结肠部位的黏膜上皮细胞1、侵袭力：微生物肠2 2、毒、毒 素：素：内毒素内毒素作用于肠壁作用于肠壁通透性增加（腹泻）通透性增加（腹泻）粘膜炎症、溃疡（脓血黏液便）粘膜炎症、溃疡（脓血黏液便）局部局部全身全身内毒素血症（发热、神志障碍、中毒性休克）内毒素血症（发热、神志障碍、中毒性休克）外毒素外毒素 志贺毒素志贺毒素植物植物NS功能紊乱（腹痛、里急后重）功能紊乱（腹痛、里急后重）干扰蛋白合成，导干扰蛋白合成，导致上皮细胞损伤致上皮细胞损伤微生物肠杆菌科572、毒素：内毒素作用于肠壁通透性增加（腹泻）粘膜炎症、溃所致疾病所致疾病细菌性痢疾细菌性痢疾急性细菌性痢疾急性细菌性痢疾急性典型：急性典型：发热、腹痛、里急后重、脓血粘液便急性中毒型：小儿常见，全身性中毒症状慢性细菌性痢疾慢性细菌性痢疾：病程迁移二月以上：病程迁移二月以上传染源传染源：病人和带菌者病人和带菌者传播途径传播途径：粪粪口传播口传播微生物肠杆菌科58所致疾病细菌性痢疾急性细菌性痢疾急性典型：发热、腹痛、里中毒性痢疾病人微生物肠杆菌科59n Specimens:Freshstool,mucusflecks,andrectalswabs.LargenumbersoffecalleukocytesandsomeRBCsmayoftenbeseenmicroscopically.n Culture:Differentialandselectivemediaasusedforsalmonellae.n Serology:Multipledrugresistance.Laboratory diagnosisTreatment:微生物肠杆菌科60Specimens:MultipledrugresiHumansaretheonlyreservoirforshigellae.Transmissionofshigellae:water,food,fingers,feces,andflies.Mostcasesoccurinchildrenunder10yearsofage.Preventionandcontrolofdysentery:1.Sanitarycontrolofwater,foodandmilk;sewagedisposal;andflycontrol.2.Isolationofpatientsanddisinfectionofexcreta.3.Detectionofsubclinicalcasesandcarriers.Prevention and control 微生物肠杆菌科61HumansaretheonlyreservoirSalmonella微生物肠杆菌科62Salmonella微生物肠杆菌科62微生物肠杆菌科63微生物肠杆菌科631.MorphologyGram-negativeRodshaped(bacilli)Mostformpiliandperitrichousflagella(周鞭毛)Donotformendospores2.Culture characteristicsFacultativeanaerobes(兼性厌氧)Growreadilyonsimplemedia.Bacterial characterization微生物肠杆菌科64MorphologyBacterialcharacteri3.Biochemical reactionsLactosefermentationtest(乳糖发酵试验):-Glucosefermentationtest:+/H2Stest:+/-Motilitytest:+4.Antigens:Oantigens,Hantigens,Viantigens微生物肠杆菌科65Biochemicalreactions微生物肠杆菌科65微生物肠杆菌科66微生物肠杆菌科66微生物肠杆菌科67微生物肠杆菌科67Salmonella on SS agar（colourless colony with a black center）微生物肠杆菌科68SalmonellaonSSagar（colourleSalmonella on SS agar微生物肠杆菌科69SalmonellaonSSagar微生物肠杆菌科69微生物肠杆菌科培训ppt课件70Motility testThe test is based on the flagellum of the bacteria.If the bacteria have flagellum,the semisolid medium will be clouding and the test will be positive.Semisolid medium微生物肠杆菌科71MotilitytestThetestisbaseSalmonellaSSagar:flaxencolonywithblackcenterKIagar:yellowslantsandbutts,nobubbleinthebottom,thelineofinoculationisblurryproduceH2SSemisolid:turbidgrowth,withflagella微生物肠杆菌科72SalmonellaSSagar:flaxencoloPathogenic substances1.Invasiveness(侵袭力)小肠黏膜（M细胞）Pili（特异性菌毛）SPI-andSPI-（型分泌系统）Acidtoleranceresponse,atr（耐酸应答基因）Oxidase（氧化酶）,SOD（超氧化物歧化酶）Viantigen（微荚膜）2.Endotoxin(内毒素)：fever,leukopenia,shock3.Enterotoxin(肠毒素):likeETECenterotoxin.微生物肠杆菌科73PathogenicsubstancesInvasiven沙门菌(红色)入侵培养的人体细胞微生物肠杆菌科74沙门菌(红色)入侵培养的人体细胞微生物肠杆菌科74Clinical findings1.Entericfever(肠热症)2.Foodpoisoning(食物中毒)：最常见最常见3.Septicemia(败血症)4.Carrierstate(无症状带菌者)微生物肠杆菌科75ClinicalfindingsEntericfever胆囊胆囊-肠道肠道-粪排菌粪排菌皮肤皮肤-血栓出血血栓出血-玫瑰疹玫瑰疹肾肾-尿尿肝脾肝脾-肿大肿大骨髓骨髓-受抑制受抑制伤寒和副伤寒的致病过程伤寒和副伤寒的致病过程伤寒和副伤寒杆菌伤寒和副伤寒杆菌小肠上部粘膜小肠上部粘膜肠系膜淋巴结肠系膜淋巴结 固有层淋巴组织固有层淋巴组织进入血液进入血液再次进入血液再次进入血液第一次菌血症第一次菌血症第二次菌血症第二次菌血症第第1周周第第23周周细胞免细胞免疫增强疫增强第第4周周微生物肠杆菌科76胆囊-肠道-粪排菌伤寒和副伤寒的致病过程玫瑰疹玫瑰疹微生物肠杆菌科77玫瑰疹微生物肠杆菌科77Typoid Mary(“伤寒玛丽伤寒玛丽”)微生物肠杆菌科78TypoidMary(“伤寒玛丽”)微生物肠杆菌科78Specimens:1stweek:peripheralblood1st3rdweeks:bonemarrow2ndweek:stool3rdweek:urineLaboratory diagnosisSS平板Biochemical reactionsSerological testH2Stest,etcAgglutinationtest微生物肠杆菌科79Specimens:LaboratorydiagnoWidal Test 肥达试验肥达试验Widal testisatubeagglutinationtestemployedintheserologicaldiagnosisofentericfever.Principle:Patientssufferingfromentericfeverwouldpossessantibodiesintheirserawhichcanreactandagglutinateserialdoublingdilutionsofkilled,colouredSalmonellaantigensinatubeagglutinationtest.微生物肠杆菌科80WidalTest肥达试验微生物肠杆菌科80Widal Test微生物肠杆菌科81WidalTest微生物肠杆菌科81肥达试验肥达试验原理原理：用已知伤寒沙门菌用已知伤寒沙门菌O、H抗原以及引起副伤寒的抗原以及引起副伤寒的甲型副伤寒沙门菌、肖氏沙门菌和希氏沙门菌甲型副伤寒沙门菌、肖氏沙门菌和希氏沙门菌H抗原的抗原的诊断菌液诊断菌液与与受检血清受检血清作试管或微孔板凝集试验，测定作试管或微孔板凝集试验，测定受检血清中有无相应抗体及其效价的试验。受检血清中有无相应抗体及其效价的试验。正常值：伤寒正常值：伤寒 O1:80，H 1:160 副伤寒副伤寒 H1:80动态观察：效价递增或恢复期效价比初次动态观察：效价递增或恢复期效价比初次4倍倍微生物肠杆菌科82肥达试验原理：用已知伤寒沙门菌O、H抗原以及引起副伤寒的甲型O O抗体与抗体与H H抗体的诊断意义：抗体的诊断意义：a)a)O O、H H凝集效价均超过正常值，肠热症可能性大。凝集效价均超过正常值，肠热症可能性大。b)b)O O、H H均低于正常值，肠热症可能性不大。均低于正常值，肠热症可能性不大。c)c)O O不高不高H H高，有可能是预防接种或非特异性回忆反应。高，有可能是预防接种或非特异性回忆反应。d)d)O O高高H H不高，感染早期或其他沙门菌感染后的交叉反应。不高，感染早期或其他沙门菌感染后的交叉反应。微生物肠杆菌科83O抗体与H抗体的诊断意义：O、H凝集效价均超过正常值，肠热症Key questionsDefinition:Widal TestDescribe the pathogenic mechanisms of ST and LT produced by E.coli.Describe the pathogenic substances of ETEC.(ST,LT,adhesins).Describe the virulent substances produced by Shigella.微生物肠杆菌科84KeyquestionsDefinition:Widal