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1.
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 总被引：1，自引：0，他引：1
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。 相似文献
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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 总被引：2，自引：0，他引：2
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。 相似文献
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猪水肿病大肠杆菌分离、鉴定及药敏试验 总被引：6，自引：3，他引：3
本实验从疑似猪水肿病的病例分离到5株大肠杆菌,O抗原鉴定结果表明所有菌株均为O139血清型;应用F18ab菌毛单克隆抗体对这5株大肠杆菌能否表达F18ab菌毛进行了鉴定,结果表明其中2个菌株能表达F18ab菌毛;利用聚合酶链式反应(PCR)对志贺氏菌样毒素Ⅱ型变异体(SLT-Ⅱe)操纵子基因保守区进行了扩增,结果发现在能表达F18ab菌毛2个菌株中可扩增一段特异性序列。以上数据表明这2株大肠杆菌为致仔猪水肿病大肠杆菌。药敏试验表明这两株菌株均对氟哌酸、妥布霉素、庆大霉素、利福平等抗生素高度敏感。 相似文献
4.
牛产肠毒素大肠杆菌毒力因子多重PCR检测方法的建立 总被引：6，自引：1，他引：6
通过多重PCR扩增产肠毒素大肠杆菌（enterotoxigentic E.coli，ETEC）的毒力因子F41菌毛、K99菌毛和STa肠毒素的编码基因来检测和鉴定ETEC。试验中对影响PCR扩增的dNTP、Mg^2＋、引物浓度以及退火温度等因素进行优化，在优化条件的基础上，确定多重PCR的特异性和灵敏性，以此建立同时检测ETEC多个毒力因子的多重PCR方法。用该方法对分离于犊牛腹泻和犊牛肠毒血症的7株大肠杆菌进行检测，结果2株为F41、K99和STa阳性，4株为F41、STa阳性，1株为K99STa阳性。这与玻片凝集试验检测菌毛的结果一致。试验表明，该方法特异性强、敏感性高、简便、快速，适用于临床鉴定和检测牛ETEC菌株。 相似文献
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牦牛肠毒素型大肠杆菌的分离鉴定和某些生物学特性的研究 总被引：1，自引：0，他引：1
从西藏地区腹泻死亡牦牛中分离出一株肠毒素型大肠杆菌并对其某些生物学特性进行了研究。结果表明，该菌在形态特`征培养特性和生化特性方面与大肠杆菌基本一致。血清学试验表明，该菌株O抗原属O148，K88、K99、987P单因子血清均不能凝集本菌；该菌不产生溶血素；对绵羊、豚鼠、马、鸡的红细胞表现强凝集，而K88、K99、987P抗血清均不能抑制其它对绵羊、豚鼠、马、鸡红细胞的凝集；该菌株在营养肉汤中经37℃，48小时培养表达菌毛；肌肉接种兔、腹腔接种小鼠均具有高致病性；乳鼠胃内投报试验和兔回肠结扎试验证明，该菌能产生热稳定肠毒素和热敏性肠毒素；分离菌株对恩诺沙星、环丙沙星、阿莫西林、羧苄青霉素等高度敏感，而对链霉素、四环素、土霉素等表现耐药性。 相似文献
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为了调查新疆某规模化奶牛场犊牛腹泻的原因并确定病原,无菌采集15份腹泻犊牛粪样进行病原的分离培养,采用形态学观察、生化试验、血清学和致病性分析等方法鉴定分离菌株,利用大肠杆菌16S rRNA通用引物进行基因序列扩增并测序,使用DNAStar软件将分离菌株测序结果与GenBank中的其他菌株序列进行同源性比对,采用Mega 6.0软件依据16S rRNA序列构建分离株系统进化树并进行分析,同时对分离菌株的黏附性基因进行鉴定。结果从15份样品中分离获得2株分离菌株;试验显示,分离株均发酵葡萄糖,MR试验、吲哚和甲基红试验呈阳性,硫化氢、氧化酶、DNA酶、VP试验呈阴性。2株分离菌株血清型均为O78,且均具有致病性。药敏试验显示,分离菌株均对环丙沙星、卡那霉素、氟哌酸、庆大霉素、磺胺甲唑、头孢哌酮、壮观霉素、多黏菌素B、呋喃妥因敏感。分离株16S rRNA基因序列与大肠杆菌菌株NF738(GenBank登录号:MT649856)同源性为≥99.7%,且处于同一大分支。经PCR鉴定,分离菌株具有K88黏附性基因。本研究结果证实了引起新疆某规模化奶牛场犊牛腹泻的病原为致病性产肠毒素型大肠杆菌。 相似文献
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肠毒素大肠杆菌((Ent erot oxi geni c E.col i,ETEC)是引起犊牛腹泻的主要病原之一。本试验建立了多重PCR检测ETEC毒力因子F41菌毛、K99菌毛和STa、LT肠毒素相关基因的技术方法。试验对影响PCR扩增的dNTP、引物浓度以及退火温度等因素进行优化,确定了多重PCR的特异性和灵敏性。结果表明:所建立的多重PCR方法快速、特异、灵敏,在2.5h-3h内就可以完成,为致犊牛腹泻肠毒素大肠杆菌的快速准确检测提供另一种选择。 相似文献
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奶牛乳腺炎是制约奶牛养殖效益和奶品质的主要疾病之一，为了解当前长春市某大型集约化奶牛场奶牛乳腺炎病原菌流行和耐药情况，本试验对该场奶牛乳腺炎发病情况进行了调查，对采集的119份临床型乳腺炎乳样进行细菌分离鉴定并对分离鉴定的病原菌进行小鼠致病性试验和药敏试验。结果显示：该场奶牛乳腺炎发病率在4%左右，初产牛乳腺炎发病率明显高于经产牛，乳腺炎发病率还与胎次、年龄等因素相关。本试验共分离鉴定出木糖葡萄球菌49株(42.2%)、松鼠葡萄球菌39株(33.6%)、大肠杆菌15株(12.9%)、金黄色葡萄球菌5株(4.3%)、无乳链球菌3株(2.6%)、菠萝泛菌3株(2.6%)、产酸克雷伯菌1株(0.9%)和铜绿假单胞菌1株(0.9%);其中菠萝泛菌、无乳链球菌和铜绿假单胞菌对小鼠具有较强致病性。药敏试验结果显示，分离菌株的耐药性普遍存在，大肠杆菌和铜绿假单胞菌耐药种类最多。除无乳链球菌外，其他菌株均对庆大霉素或阿米卡星敏感，可作为该场奶牛乳腺炎治疗首选药物。本试验通过对该场乳腺炎调查、分离菌鉴定和耐药性分析，为该场乳腺炎的防治提供了参考依据。 相似文献
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乳房炎是奶牛养殖业中最常见、防治最困难、花费最多的疾病之一,给奶牛养殖业带来了巨大的经济损失,本实验大肠杆菌是奶牛乳房炎的主要致病菌.本试验旨在对新疆昌吉州及周边地区规模化奶牛场的乳房炎奶牛乳汁进行细菌分离培养与鉴定,调查其中的大肠杆菌在奶牛场的分布情况;同时,进行该菌的药敏试验和耐药检测,获得大肠杆菌对哪种抗生素敏耐药并提出合理的预防措施。无菌采集奶样,冰盒保存,12h内送到实验室进行病原菌的分离鉴定,对大肠杆菌进行药敏试验和耐药监测。结论:通过分离鉴定的大肠杆菌,预期测定对大肠杆菌敏感的药物,对牛场防治奶牛乳房炎提供的重要线索。 相似文献
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Protection against enteric colibacillosis in pigs suckling orally vaccinated dams: evidence for pili as protective antigens 总被引：8，自引：0，他引：8
H W Moon 《American journal of veterinary research》1981,42(2):173-177
Pregnant gilts were vaccinated orally with Escherichia coli that produced pilus antigens K99 or 987P. The vaccines were live or dead enterotoxigenic E coli (ETEC) or a liver rough non-ETEC strain which has little ability to colonize pig intestine. Pigs born to the gilts were challenge exposed orally with K99+ or 987P+ ETEC, which did not produce heat-labile enterotoxin or flagella and which produced somatic and capsular antigens different from those of the vaccine strains. Control gilts had low titers of serum and colostral antibodies against pilus antigens, and their suckling pigs frequently had fatal diarrhea after challenge exposure. Serum antibody titers against pilus antigens of the vaccine strains increased in the gilts after vaccination with liver ETEC, and the colostral antibody titers of these gilts were higher than those of controls. Pigs suckling such vaccinated gilts were more resistant than controls to challenge strains were of different pilus types, and it could not be attributed to enterotoxin neutralization by colostrum. In contrast to the live ETEC vaccines given to the pregnant gilts, the liver rough non-ETEC and dead ETEC vaccines stimulated little or no production of antibody against pilu, and the pigs born of these vaccinated gilts remained highly susceptible to challenge exposure. The results support the hypothesis that pilu can be protective antigens in oral ETEC vaccines. It was indicated that in the system reported, protection depended on living bacteria for the production of pilus antigens in vivo or for the transport of pilus antigens across intestinal epithelium. 相似文献
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The pathogenicity of Yersinia enterocolitica strains isolated from various sources in four test systems. 总被引：1，自引：0，他引：1 
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下载免费PDF全文 Six tests were applied to 39 strains of Yersinia enterocolitica of various serotypes and from several sources in an attempt to relate the test to pathogenicity of the strains. The tests that were used were the pig gut loop test and the infant mouse test for heat stable enterotoxin, the Sereny and HeLa cell tests for invasiveness, inhibition of growth on magnesium oxalate agar, and the ability to cause diarrhea in infant mice. The pig gut loop test was found to be unsuitable for detection of heat stable enterotoxin but 20 strains produced heat stable enterotoxin that was detected in infant mice. None of the strains was positive in the Sereny test but 21 invaded HeLa cells. The growth of 20 strains was inhibited at 37 degrees C on magnesium oxalate agar and, in the orally-infected mice, 23 strains caused diarrhea or death. These findings indicate a discrepancy between the infant mouse test and the ligated intestine test in pigs for heat stable enterotoxin and a significant difference in Y. enterocolitica heat stable enterotoxin compared with Escherichia coli heat stable enterotoxin because the former failed to elicit a significant response in pig intestine. 相似文献
13.
R E Holland N Sriranganathan L DuPont 《Journal of the American Veterinary Medical Association》1989,194(3):389-391
Enterotoxigenic Escherichia coli was isolated from a 3-day-old foal with diarrhea. The isolate was distinguished from nonpathogenic E coli by determining the presence of pili and enterotoxin production. A standard slide agglutination test was performed, using pooled antisera that contained antibodies against K99 and F41 pilus antigens, K87 capsular antigen, and 0101 somatic antigen. Agglutination of the antisera occurred in the presence of the isolate. Piliation was verified by use of negative-contrast electron microscopy. Further, the isolate produced a heat-labile enterotoxin-like antigen that cross-reacted with a reagent containing formalin-treated, heat-killed Staphylococcus aureus (cowan 1 strain) bearing anti-cholera antibodies. On the basis of the aforementioned procedures and the absence of other identifiable enteric pathogens, we believe that E coli was responsible for causing diarrhea in the foal. 相似文献
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D García M E Cavazza L Botero M Gorziglia G Urbina F Liprandi J Esparza 《Veterinary microbiology》1987,13(1):47-56
The aetiology of neonatal porcine diarrhoea was studied in 15 different herds located in the north-western region of Venezuela. Of 56 strains of Escherichia coli analyzed, 16 (28.6%) were shown to produce heat-stable (STa) enterotoxin, as detected by infant mouse assay. Only four of these STa+ isolates also possessed the K88 pilus antigen, two were 987P+ and none possessed the K99 antigen, leaving 10 STa+ samples in which no pilus antigen was identified. Among the 40 STa negative samples were six K88+ specimens, one K99+, four 987P+, one which reacted as K88+ + K99+ and one K88+ + 987P+. Considering as pathogenic any strain showing at least one of the characters studied, pathogenic E. coli were detected with an overall frequency of 42.9%, being more prevalent during the second week of life. An electrophoretic analysis of the plasmid content of the field isolates of E. coli, revealed the presence of numerous species of extrachromosomal DNA, although no direction association could be made between a particular plasmid and any of the pathogenic characteristics identified. Results of Southern blot analysis indicate that the STa enterotoxin was preferentially encoded within an endemic plasmid of 4.9 Md. Other plasmids present in the E. coli isolates could be related to antibiotic resistance. With the exception of one strain, all E. coli isolates were resistant to more than one of the nine drugs tested; multiresistant E. coli were frequently isolated, including four strains which were resistant to seven antibiotics. 相似文献
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Prevalence of enterotoxigenic Escherichia coli in calves in Scotland and northern England 总被引：1，自引：0，他引：1
Eighty-eight of 1529 (5.7 per cent) Escherichia coli isolates from diarrhoeic and clinically normal calves in Scotland and northern England were found to possess the K99 pilus antigen (K99+). There was complete correlation between possession of K99 antigen, heat stable enterotoxin production and ability to dilate intestinal loops. The diagnosis of calf enterotoxigenic E coli infections may therefore be based on the detection of K99 antigen alone. Enterotoxigenic E coli was isolated from 23 of 306 (7.5 per cent) diarrhoeic calves from eight of 70 (11.4 per cent) farms and was not isolated from clinically normal calves. Infected calves were between one and three days old. A survey by an enzyme-linked immunosorbent assay found 3.0 per cent and 3.9 per cent of sera from calves and cows respectively to contain antibodies to K99 antigen. The prevalence of other enteropathogenic organisms in calf faeces is also discussed. 相似文献
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L Niilo 《Research in veterinary science》1976,20(2):225-226
Lethal doses of enterotoxin of Clostridium welchii (perfringens) type A injected intravenously into young fowls caused immediate lassitude, with partial recovery, followed by death seven to 35 h after inoculation. Lesions found were ascites, hydropericardium, oedema of the muscles, hepatic congestion, urate deposits on the peritoneum and the pericardium, and intestinal hyperaemia. Sublethal doses produced no clinical signs or lesion. The LD50 of this enterotoxin was 74-84 mug/kg of body weight. 相似文献
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Use of immunofluorescence, Gram's staining, histologic examination, and seroagglutination in the diagnosis of porcine colibacillosis 总被引：3，自引：0，他引：3
D H Francis 《American journal of veterinary research》1983,44(10):1884-1888
An indirect fluorescent antibody (IFA) test of ileal impression smears for K88, K99, and 987P pilus antigens was compared with histologic examination and seroagglutination of Escherichia coli isolates for efficacy in determining colibacillosis in pigs. Histologic examination appeared to be more effective than the IFA test in revealing colonization of the ileum by bacteria. However, histologic examination revealed little about the nature of the colonizing bacteria. Correlation between bacterial adherence, as observed in histologic sections of ileum, and the presence of bacteria with adherence pili, as determined by IFA testing, was 91%. Results of seroagglutination for pilus antigens correlated with results of histologic examination in only 84% of the cases. Pilus antigens were not identified by IFA testing or seroagglutination in 5% of the cases in which adherent bacteria were observed in histologic sections of the small intestine. Because little preparation time was required for the IFA test, results were available within 2 hours of necropsy. In contrast to histologic examination, the IFA test made possible identification of the colonizing organism as E coli and revealed the type of pilus antigen present. Adherence pili were identified more frequently by IFA testing than by seroagglutination. Examination of Gram's-stained ileal impression smears was useful in screening for colibacillosis in pigs. Bacterial adherence was found in positive correlation with the number of gram-negative bacilli observed. Bacterial adherence rarely was observed in histologic sections of ileum when smears contained less than or equal to 10 gram-negative bacilli/1,000 X microscopic field. Each diagnostic test compared offered advantages and disadvantages over the other tests. Seemingly, concurrent use of several of these tests, rather than one, should be used in the diagnosis of porcine colibacillosis. 相似文献
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Influence of bovine intestinal fluid on the expression of K99 pili by Escherichia coli 总被引：2，自引：0，他引：2
A study was conducted to determine whether intestinal fluid collected from various portions of bovine intestine differed in its effect on production of K99 pili by Escherichia coli. The small and large intestines of 7 calves, euthanatized 4 hours after a final feeding of milk, were divided into 6 to 9 segments from which intraluminal fluids were collected. Depending on the amount of fluid collected, up to 20 E coli strains that express K99 pili were grown on media prepared from the content of each specimen and then were tested for K99 pilus expression. In general, intestinal fluid from the most proximal small intestinal segments were more suppressive to K99 pilus expression than was fluid from more distal segments of small intestine. Only about 20% of the E coli test strains expressed K99 pili when grown on medium prepared from proximal small intestinal segmental fluid, whereas greater than 90% did when grown on medium prepared from distal small intestinal segmental fluid. Fluid from the large intestine varied considerably from calf to calf in its effect on K99 pilus expression. A correlation was found between K99 pilus expression and pH of the intestinal fluid, with the lower pH values (characteristic of proximal intestinal segmental fluid) being suppressive. The correlation between K99 pilus production and the pH of the medium was verified, using defined laboratory media adjusted to various pH values. Strains of E coli grown in medium at or below pH 5.5 failed to express K99 pili, whereas the same strains when grown in medium at or above pH 6.5 expressed K99 pili in abundance. 相似文献