重庆市致病性大肠杆菌的血清型鉴定

从重庆市各地发生典型仔猪黄、白痢猪的肛拭粪样及死亡猪的心血和肝脏中分离鉴定出173株致病性大肠杆菌,用20种大肠杆菌O抗原单价血清进行了血清学鉴定。结果表明:173株分离菌有87株能定型,分属14种血清型,其中以O101(17.2%)、O8(14.9%)、O20(14.9%)、O64(11.5%)、O45(9.2%)、O149(8%)等6种为优势血清型。同一地区存在多种血清型,大多数地区有本地的优势血清型。不同地区的优势血清型有的相同,有的不同。     

青海省部分仔猪水肿病病原的血清型鉴定

就青海省仔猪水肿病多发区分离鉴定的45株溶血性大肠杆菌进行了血清学定型，结果表明：已鉴定出血清型的菌株33株，占73．3％，12株未定出血清型，占26．7％；其中O8 10株(22．2％)，O25 1株(2．2％)，O101 1株(2．2％)，O138 3株(6．7％)，O139 7株(15．5％)，O141 5株(11．1％)，O149 6株(13．3％)。O抗原优势菌株分别为：O8、O139、O141、O139，占全部菌数的62．2％。     

温州地区鸡肠道大肠埃希菌耐药性分析及血清型鉴定

为研究温州地区鸡肠道大肠埃希菌的耐药性和血清型,从27家养鸡场采集泄殖腔拭子355份,分离并鉴定出346株鸡大肠埃希菌,分离率为97.46%。在346株分离株中,对3种以上抗菌药物耐药的菌株占84.39%(292/346),呈现多重耐药性。O血清型鉴定结果显示,分离菌株分别属于13个血清型,其中O138、O146、O136、O45占分离菌株的39.02%(135/346),为优势血清型。     

宁夏部分地区鸡大肠埃希菌的分离与血清型鉴定

为调查宁夏地区鸡大肠杆菌病的流行情况,采集宁夏4个地区的103个规模化鸡场疑似病料151份,通过病原菌分离鉴定及生化试验,鉴定出鸡大肠埃希菌109株,平均分离率为72.19%(109/151);动物试验证实,59株分离菌有致病性,占分离菌54.13%(59/109);对致病性菌株进行血清型鉴定,确定血清型种类13种38株,占致病性菌株64.41%(38/59);优势血清型分别为O15、O18、O78、O88,占定型菌株63.16%(24/38)。     

安徽部分地区猪场产肠毒素大肠杆菌的分离鉴定及其生物学特性分析

产肠毒素大肠杆菌(enterotoxigenic Escherichia coli,ETEC)是猪场仔猪黄白痢的重要病原之一。为调查安徽省猪场中ETEC的感染情况,在2018年对该省6个地区的17个规模化猪场的仔猪黄白痢样品进行了ETEC的分离鉴定。结果:经PCR鉴定,在479份病料中分离出110株ETEC菌株。对这些菌株进行血清型、耐药性和致病力分析表明,该110株ETEC菌株共鉴定出13种血清型,以O8、O20、O128为优势血清型。110株ETEC菌株表现出高耐药性,对强力霉素的耐药率高达100%(110/110);对新霉素、红霉素、庆大霉素、阿莫西林、四环素等抗生素的耐药率均高达70%以上,且多数菌株表现出多重耐药,其中9、10重耐药菌株数最多。分别将优势血清型O8、O20、O128菌株对小鼠进行攻毒,结果同一血清型中强毒株数量分别占该血清型菌株总数的50%(55/110)、72.7%(8/11)和66.7%(4/6),表明这些优势血清型菌株的毒力普遍较高。本研究为猪场ETEC引起的仔猪黄白痢的流行病学研究及防控提供一定的参考。     

衡水地区奶牛乳房炎源大肠杆菌分离鉴定、血清型及耐药性分析

为了确定衡水地区奶牛乳房炎源大肠杆菌(E.coli)的血清型及耐药性,从衡水地区不同的奶牛场采集60份患乳房炎奶牛奶样,分离鉴定出20株E.coli;经血清型鉴定共有8种血清型,以O9和O13为优势血清型;耐药性分析结果表明20株E.coli分离株仅对头孢克肟、恩诺沙星、丁胺卡那、强力霉素4种药物敏感,对其他药物存在不同程度的耐药性。     

致奶牛乳腺炎大肠杆菌的分离与鉴定

从我国7个省(市、自治区)部分地区的奶牛场及养殖小区采集临床显性和隐性乳腺炎乳样病例530份,从中分离、鉴定出大肠杆菌95株,分离率为17.9%。通过玻板凝集和试管凝集试验结果表明,95个大肠杆菌分离株中,有54个分离株被鉴定出37种血清型,另有2株自凝,39株未定型,分别占检验菌株的56.85%、2.1%和41.05%。54个已定型菌株中,常见血清型为O93、O9、O146、O7、O74,分别有8、6、5、3、3株,分别占定型菌株的14.81%、11.11%、9.26%、5.55%、5.55%。且部分分离菌株可与多种单因子血清发生凝集反应,从而表现出多种血清型。     

奶牛粪样中致病性大肠埃希菌的分离与鉴定

大肠埃希菌血清型众多,一些具有特殊血清型的大肠埃希菌对人和动物有致病性。本试验从包头市侯家营奶牛场77头外表健康成年泌乳荷斯坦奶牛采集新鲜粪样,进行细菌分离培养,共分离到疑似大肠埃希菌69株,经过形态观察、生化鉴定、动物致病性试验和血清型鉴定,其中12株为致病性大肠埃希菌,均为强毒株,占分离菌数的17.4%,另57株为非致病性大肠埃希菌,占分离菌数的82.6%。同时对12株致病性大肠埃希菌进行了"O"血清型鉴定,分属7种血清型O78、O6、O9、O98、O1、O8、O149,分别占定型菌株的25.0%(3/12)、8.3%(1/12)、16.7%(2/12)、8.3%(1/12)、8.3%(1/12)、8.3%(1/12)、25.0%(3/12)。     

海安县鸡大肠杆菌病病原的分离鉴定及多价灭活苗的研制

为了确定海安县鸡大肠杆菌病的病原并研制多价灭活苗,试验对从海安县兽医站门诊及12个大型养殖场具有疑似大肠杆菌病病变的252只病死鸡进行了病原分离及鉴定。结果表明:共分离、鉴定出大肠杆菌226株,定型198株;共测出23个血清型,其中O78(32株)、O18(26株)、O2(21株)、O88(17株)、O1(13株)、O11(10株)6种血清型占定型菌株的60.1%,为海安县大肠杆菌优势血清型;将此6种优势血清型大肠杆菌制成多价灭活苗免疫雏鸡,安全性好,保护率高。     

临沂市鸡源致病性大肠杆菌的优势血清型监测及其致病性试验

为了查清我市禽大肠杆菌的血清型及其致病性,对我市不同区域的23个规模化养鸡场及60个养鸡户的412份疑似大肠杆菌病病例进行了细菌分离,并对其进行了生化试验、致病性试验、血清学鉴定。结果:(1)412份病料中分离出了362株大肠杆菌,分离率为87.9%;⑵不同区域的鸡场流行的血清型有较大差异,在362个分离株中,除了27株未能定型、12株自凝外,共鉴定出323个分离株的O型血清型,占分离株的89.2%,这些定型菌株覆盖了36个血清型;(3)优势血清型为O1、O78、O18、O11、O88、O24,这6种血清型共占定型菌株的56.3%;⑷6种优势血清型均具有致病性,其中O1、O78、O18、O11、O24为高致病性菌株,O88为中致病性菌株。     

华中地区鸡源沙门菌分离鉴定及耐药性分析

为了探明华中地区种鸡场沙门菌(Salmonella)的优势血清型和耐药情况,本研究从湖北、河南、湖南等省市22个规模化鸡场采集病鸡、死胚及弱雏组织样品3 724份,通过分离培养、生化试验、PCR鉴定及血清型试验确定分离菌种属及其血清型,并采用Kirby-Bauer法对分离菌株进行了耐药性分析。结果显示,本试验从3 724份病料中共分离鉴定出124株沙门菌,其中79株为D群肠炎沙门菌(63.71%,79/124),34株为D群鸡白痢沙门菌(27.42%,34/124),8株为B群鼠伤寒沙门菌(6.45%,8/124),有3株沙门菌未能确定血清型。O抗原鉴定79株肠炎沙门菌和34株鸡白痢沙门菌为O9,8株鼠伤寒沙门菌为O4。H抗原鉴定79株肠炎沙门菌为Hg,m,8株鼠伤寒沙门菌为Hi。药敏试验结果显示,124株分离菌株对萘啶酸、氨苄青霉素、四环素和多西环素耐药率分别为95.97%(119/124)、91.94%(114/124)、57.26%(71/124)和70.16%(87/124);对复方新诺明和红霉素耐药率分别为25.81%(32/124)和12.10%(15/124);对氯霉素、庆大霉素、头孢噻肟和卡那霉素耐药率分别为6.45%(8/124)、1.61%(2/124)、1.61%(2/124)和0.81%(1/124);对左氧氟沙星、阿米卡星和多黏菌素B完全敏感。99.19%(123/124)的分离株至少对一种药物耐药,87.10%(108/124)的分离株表现多重耐药。本研究为华中地区养鸡场沙门菌的诊断及防控提供了数据支撑。     

新疆北疆地区致犊牛腹泻大肠杆菌的分离鉴定及部分生物学特性

为研究确定新疆北疆地区规模化奶牛场犊牛腹泻病原性大肠杆菌的优势血清型、致病性及毒力因子特征。采用细菌学、免疫学及分子生物学的方法对从新疆呼图壁、石河子、奎屯3个主要奶牛生产基地14个规模化奶牛场10日龄内腹泻犊牛直肠棉拭子样品进行了大肠杆菌的分离与鉴定、O血清群、黏附素及肠毒素的测定。结果从302份样品中分离并经生化鉴定获得180株呈β溶血的大肠埃希菌,其中94株对小鼠有致病性;对其中53株代表菌株的O血清型、黏附素及肠毒素测定,结果为8株携带K99菌毛及STa毒素基因,6株携带F41茵毛基因,6株产生LT毒素;28株分离株分布于16个O血清型,其中O101,O6,O114,O78为优势血清型,占被测菌株的50％。结果表明,新疆北疆主要奶牛养殖区致犊牛腹泻大肠杆菌是以携带K99、F41和产ST的溶血性大肠杆菌为主,其研究结果为犊牛大肠杆菌性腹泻的免疫防治提供病原学依据。     

动物性食品源大肠杆菌O血清型鉴定及其K88菌毛基因检测

本研究对河北省冀东地区农贸市场和超市采集的生猪肉、生鸡蛋和生羊肉等分离得到的20株大肠杆菌进行大肠杆菌血清型鉴定;并检测不同血清型大肠杆菌的K88菌毛基因。采用常规方法进行大肠杆菌的O血清型鉴定,用PCR方法检测K88菌毛基因。分离鉴定的20株大肠杆菌有7种血清型,包括O38、O78、O88、O11、O107、O91、O9,其中O38、O78为优势血清型菌,均占分离菌株的25%(5/20)。在分离的动物性食品源大肠杆菌中有30%(6/20)的菌株K88菌毛基因扩增呈阳性。结果表明,O78、O38为冀东地区动物性食品源大肠杆菌常见血清型,30%(6/20)的菌株K88菌毛基因扩增阳性。     

仔猪腹泻大肠埃希菌的分离鉴定及药敏试验

为了了解甘肃省武威市凉州区部分地区引起仔猪黄白痢的致病性大肠埃希菌的主要血清型及其致病性,从凉州区4家规模化猪场采集了60份仔猪肛门拭子,用细菌分离培养方法分离出20株大肠埃希菌菌株,然后经形态观察、生化试验,确定了该次分离菌株为致病性大肠埃希菌。经血清型鉴定,表明20株分离菌株分布在8个血清型中,其中优势血清型为O8（9株）,占总数的45%。药物敏感性试验结果表明,分离菌株对临床常用药物具有抗药性。该研究能够对凉州区仔猪腹泻病的流行病学研究、诊断和免疫防制提供科学参考。     

Serotypes,phage types and virulence genes of shiga-producing Escherichia coli isolated from sheep in Spain

PROBLEM ADDRESSED: Shiga toxin-producing Escherichia coli (STEC), have emerged as food poisoning pathogens which can cause severe diseases in humans. OBJECTIVE: The aim of this study was to determinate the serotypes and virulence genes of STEC strains isolated from sheep in Spain, with the purpose of determining whether sheep represent a potential source of STEC pathogenic for humans. METHODS AND APPROACH: Faecal swabs obtained from 697 healthy lambs on 35 flocks in Spain during the years 2000 and 2001 were examined for STEC using phenotypic (Vero cells) and genotypic (PCR) methods. RESULTS: STEC O157:H7 strains were isolated from seven (1%) animals in six flocks, whereas non-O157 STEC strains were isolated from 246 (35%) lambs in 33 flocks. A total of 253 ovine STEC strains were identified in this study. PCR showed that 110 (43%) strains carried stx(1) genes, 10 (4%) possessed stx(2) genes and 133 (53%) both stx(1) and stx(2). Enterohaemolysin (ehxA) and intimin (eae) virulence genes were detected in 120 (47%) and in 9 (4%) of the STEC strains. STEC strains belonged to 22 O serogroups and 44 O:H serotypes. However, 70% were of one of these six serogroups (O6, O91, O117, O128, O146, O166) and 71% belonged to only nine serotypes (O6:H10, O76:H19, O91:H-, O117:H-, O128:H-, O128:H2, O146:H21, O157:H7, O166:H28). A total of 10 new O:H serotypes not previously reported in STEC strains were found in this study. Seven strains of serotype O157:H7 possessed intimin type gamma1, and two strains of serotype O156:H- had the new intimin zeta. STEC O157:H7 strains were phage types 54 (four strains), 34 (two strains) and 14 (one strain). CONCLUSIONS: This study confirms that healthy sheep are a major reservoir of STEC pathogenic for humans. However, because the eae gene is present only in a very small proportion of ovine non-O157 STEC, most ovine strains may be less pathogenic.     

Detection of Serotypes and Virulence Factors of Escherichia coli from Piglets in Beijing

To investigate the cause of piglets diarrhea, and the distribution of the serotype and virulence factors of swine Escherichia coli in Beijing, 400 diarrhea samples were collected. TSA serum agar culture method was used to isolate Escherichia coli, serotype identification test and virulence factor genes test were used to verify the presence of O-antigen. 64 strains of E.coli were isolated from 400 diarrhea samples, among which 42 strains of E.coli were classified as 8 serotypes:O101(18.7%),O64(12.5%),O8(10.9%),O20(10.9%),O45(4.7%),O149(4.7%),O2(1.6%) and O89(1.6%), and the major virulence factors were STa, Stx2e, astA and eaeA. There were 8 mainly serotypes that caused piglets diarrhea in Beijing area, among which O101 serotype accounted for the highest proportion. The major virulence factors were astA and eaeA, accounted for more than 50% of all strains, STa and Stx2e accounted for more than 30% of all strains. These data would provide effective data to support the prevention and control of piglet diarrhea in Beijing.     

北京地区仔猪大肠杆菌血清型鉴定及其毒力因子检测

本试验旨在调查北京地区仔猪腹泻的原因及猪源大肠杆菌血清型、毒力因子的分布情况。收集北京京郊地区仔猪腹泻样本400份,应用含2%血清的TSA培养基分离大肠杆菌,通过血清型鉴定试验和毒力因子检测验证O抗原。结果显示,400份样本中分离到64株大肠杆菌,定型42株。其主要为O101型（18.7%）、O64型（12.5%）、O8型（10.9%）、O20型（10.9%）、O45型（4.7%）、O149型（4.7%）、O2型（1.6%）、O89型（1.6%）8种血清型,其携带的毒力因子为STa、Stx2e、astA和eaeA等。结果表明,引起京郊仔猪腹泻的大肠杆菌主要有8个血清型,其中O101型发病比例最高。其携带的毒力因子主要为astA和eaeA,占全部菌株的50%以上,STa和Stx2e因子占全部菌株的30%以上。本试验结果将为今后京郊地区仔猪腹泻病的防控提供有效的数据支持。     

Phenotypic characterization of ipaH+ Escherichia coli strains associated with yolk sac infection

Seventy-six Escherichia coli serotypes possessing the ipaH gene typical of enteroinvasive E. coli (EIEC) strains were characterized. Biochemical identification of our strains shows positive reactions for lactose fermentation (100% of strains), lysine decarboxylase (98.7% of strains) and motility (67.1% of strains), properties that do not correspond with those described to the EIEC group. The serotypes agree with an initial classification. In this, some common O antigens identified among ipaH+ strains were O2 (n=20), OR (n=11) and non-determined O? (n=10). The O2:NM serotype was the most common. Sixty-six percent (n=50) of the ipaH+ E. coli strains were colicin producers, of them, 26 (34%) produced Col V and other colicins, 13 (17%) produced colicins other than Col V, and 11 (14.5%) produced Col V only. Trimethoprim/Sulfa (72%), ampicillin (64.5%), enrofloxacin (55.3%), and ciprofloxacin (47.4%) were the major antimicrobial resistance frequencies observed. Twenty-five different multiresistance patterns were observed, where sixty-six strains (86.8%) were included. A MIC test showed that most of the strains were sensitive to low gentamicin and kanamycin concentrations, whereas most of the strains were resistant to tetracycline. An invasiveness assay showed that the predominant alterations caused to HEp-2 cells were changes in shape and staining, and in most of the specimens, a partial monolayer detachment was also seen. Fifteen strains invaded more than 30% of the monolayer cells, causing the formation of intercellular bridges or filipoidal-like protrusions. The results suggest the existence of specific clone complexes derived from EIEC strains adapted to the avian host. To our knowledge, this is the first study that demonstrates the presence of extraintestinal invasive E. coli (ExIEC) strains.