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1.
Serotypes of E. coli strains isolated from piglets, which died with symptoms of diarrhea in 9 swine industrial farms, were determined. Large numbers of serotypes (from 16 to 27) in individual farms were detected. The sets of serotypes from 9 investigated farms differed among each other significantly, depending on the farm and time of examination. It was found that more than one serotype of E. coli may exist in the pig body and contribute to the development of disease. The predominant serotypes, i.e. those comprising more than 10% of serologically determined strains, were found to exist in 6 of the investigated farms and not in the remaining ones. Among the predominant serotypes, particularly important seem to be strains with K88 antigen. For prophylaxis of piglet colibacteriosis in industrial farms in Poland two vaccines for sows are recommended: one containing the K88 antigen only and the other the following serotypes: 0149:K91,K88; 020:K57; 020:K83; 0157:K88; 01:K1; 0136:K78; 024:K?; 078:K80 and 0118:K? Strains belonging to these serotypes were the most prevalent in our strain collection.  相似文献   

2.
Twenty four, 21-d-old female pigs were fed diets containing either skim milk powder (CON+), spray-dried bovine colostrum (7.5%, BC+) or spray-dried bovine plasma (7.5%, BP+) and were dosed orally with 1 × 109 CFU of E. coli O149:K88. Another group of 8 unchallenged pigs was fed the skim milk powder diet (CON−). On d 19 of the experiment all piglets were euthanased. Adverse effects of the E. coli challenge were observed variously throughout the small intestine in pigs consuming either the BC+ or BP+ diets. In this experiment, similar responses to the E. coli challenge were observed in both plasma and colostrum fed pigs, which suggests that spray-dried bovine colostrum may be a potential alternative to spray-dried bovine plasma.  相似文献   

3.
旨在了解新疆地区腹泻仔猪源大肠杆菌的系统进化分群、血清型及耐药性。本研究对154份腹泻仔猪粪便样品进行大肠杆菌的分离鉴定,采用多重PCR方法对分离株进行系统进化分群和O血清型鉴定,通过K-B纸片法对其进行药物敏感性检测并通过PCR方法进行耐药基因检测。结果显示:共分离到154株大肠杆菌,包括ETEC(n=24)、STEC(n=21)、EPEC(n=1)、EPEC/STEC(n=2)、ETEC/STEC(n=1)和ETEC/EPEC(n=1),其他104株。系统进化分群显示,多数菌株属于B1(37%)和A群(31%)。定型菌株44株,分别属于10种血清型,以O154、O12、O8、O141和O175为主要流行血清型。151株(98%)为多重耐药菌,对复方新诺明、四环素、氨苄西林、链霉素和氯霉素的耐药率为81%~100%,对阿莫西林/克拉维酸、头孢噻肟、庆大霉素、头孢曲松、环丙沙星和阿米卡星的耐药率为31%~66%,对左氧氟沙星、多黏菌素B、头孢他啶、头孢吡肟、氨苄西林-舒巴坦、哌拉西林-他唑巴坦和亚胺培南的耐药率为1%~19%。耐药基因tetA(88%)、tetG(60%)和cmlA(4...  相似文献   

4.
1125 and 1146 E. coli strains isolated from suckling and weaned piglets with diarrhea, respectively, and 724 strains from healthy piglets were tested for the presence of fibriae and production of enterotoxins. The fimbriae were determined by hemagglutination and slide agglutination tests, enterotoxins—by the use of ileal loop test in piglets (LT and STb enterotoxins) and suckling mouse assay (STa enterotoxin). It was found that 72.8 and 53.0% strains, isolated from diseased suckling and weaned piglets, respectively, possessed specific fimbrial hemagglutinins, in most cases with K88 antigen. Additionally, 987P fimbriae were detected in 14.0 and 0.7% strains isolated from piglets with diarrhea. Only 5 strains (0.7%) recovered from healthy piglets had specific fimbriae, usually with undetermined antigenic structure. F1 fimbriae (called common or unspecific) were found in strains isolated both from diseased (15.2 and 16.3% strains, respectively) and healthy piglets (27.1% strains). It was noted that the strains isolated from suckling and weaned piglets with diarrhea in most cases were enterotoxigenic (90.5 and 69.1% strains, respectively) and most frequently produced heat-labile toxin LT alone or with STb. 18.5% of enterotoxigenic strains isolated from healthy piglets produced STa toxin.  相似文献   

5.
Post-weaning diarrhoea and oedema disease in weaned piglets are caused by infection with F4+ or F18+ Escherichia coli strains. There is no commercial vaccine available, but it is shown that oral immunization of weaned piglets with purified F4 fimbriae induces a protective mucosal immune response. In the present study, piglets were orally and nasally immunized with purified F18 fimbriae in the presence of the mucosal adjuvant LT(R192G) or CTA1-DD, respectively. This immunization could not lead to protection against F18+ E. coli infection. The induced F18-specific immune response was directed towards the major subunit FedA and weakly towards the adhesive subunit FedF. The results of these experiments demonstrate that it is difficult to induce protective immunity against F18+ E. coli using the whole fimbriae due to the low response against the adhesin.  相似文献   

6.
In order to evaluate the role of the AIDA-I of porcine diarrheagenic Escherichia coli strain PD20 serogroup O143 (AIDA-I+, STb+), a mutant strain PD20M (AIDA-I, STb+) was generated from strain PD20 by an allelic exchange procedure. In addition, the full-length aidA gene was reintroduced into strain PD20M to generate the complemented strain PD20C (pTaidA, AIDA-I+, STb+). A non-pathogenic E. coli strain PD71 was used as negative control. Each strain was inoculated to newborn pigs via stomach tube. Severity of diarrhea was evaluated clinically and intestinal colonization was assessed by histology, immunohistochemistry (IHC), and transmission electron microscopy (TEM) including immunogold electron microscopy (IGEM). The adhesion pattern to HeLa cells, bacterial auto-aggregation and biofilm formation were evaluated in vitro. Pigs infected with strains PD20 or PD20C developed diarrhea 16 and 28 h after inoculation, respectively, in contrast to pigs infected with strains PD20M or PD71. Histology, IHC, TEM and IGEM examinations showed heavy bacterial colonization with biofilm formation in the large intestine, and marked in vivo expression of AIDA-I protein in pigs infected with strains PD20 or PD20C in contrast to pigs infected with strains PD20M or PD71. The in vitro assays showed marked diffuse adherence to HeLa cells, enhanced bacterial auto-aggregation and significant biofilm formation (p < 0.05) by the AIDA-I+ strains, when compared to AIDA-I strains. These results demonstrate that expression of AIDA-I is essential for intestinal colonization and in vitro bacterial autoaggregation and biofilm formation. Thus, AIDA-I may be considered a significant virulence determinant in development of diarrhea caused by porcine diarrheagenic AIDA-I+ E. coli PD20 in piglets.  相似文献   

7.
为进一步完善中国大肠杆菌菌体微量凝集试验定型血清库,改良微量凝集试验用大肠杆菌O抗原定型血清的制备工艺,本试验选取大肠杆菌O50、O60、O117、O131血清型进行定型血清制备方法的探究。首先用大肠杆菌不同血清型的参考菌株制备灭活抗原,而后多次免疫家兔以采血获得粗血清,用微量凝集试验的方法确定不同粗血清的交叉凝集素及凝集效价,再通过交叉凝集素吸收法结合血清稀释法消除非特异性凝集,最终研制出特异性良好的微量凝集试验用大肠杆菌O抗原定型血清。本研究确定了大肠杆菌O50、O60、O117、O131定型粗血清的主要交叉凝集素,最终成功研制出特异性良好的微量凝集试验用大肠杆菌O50、O60、O117、O131定型血清共4种,凝集效价在1:256至1:2 048之间,其中微量凝集试验用大肠杆菌O50、O60定型血清无交叉凝集素,为单因子定型血清,微量凝集试验用大肠杆菌O117、O131定型血清存在1~2种非特异性的交叉凝集素O14和O107。本研究作为大肠杆菌O抗原定型血清制备工艺摸索过程中的重要部分,为完善中国大肠杆菌菌体微量凝集试验定型血清库,进一步改良微量凝集试验用大肠杆菌O抗原定型血清的制备方法提供了重要理论依据。  相似文献   

8.
To investigate the relationship between the mRNA expression level of m6A demethylase and E.coli F18 resistance in piglets,Real-time quantitative PCR was used to detect the mRNA expression differences of m6A demethylase FTO and ALKBH5 genes in the duodenum and jejunum tissues of E.coli F18-resistant and -sensitive individuals from 35-day Sutai weaned piglets.In E.coli F18ab,F18ac bacteria-stimulated and endotoxin LPS-induced porcine small intestinal epithelial cells (IPEC-J2),the expression levels of FTO and ALKBH5 genes were detected,respectively.The results showed that the expression levels of FTO and ALKBH5 genes in the duodenum and jejunum of resistant individuals were extremely significantly or significantly higher than those of sensitive individuals (P<0.01,P<0.05),and the expression of FTO gene were not significantly changed in E.coli F18 bacteria-stimulated IPEC-J2 cells (P>0.05),but the expression levels of ALKBH5 gene were significantly up-regulated after F18ac stimulation (P<0.05).After LPS induction for 4 hours,the expression levels of FTO and ALKBH5 genes showed significant up-regulation in IPEC-J2 cells (P<0.05).This study preliminarily verified and indicated that the expression levels of m6A demethylases FTO and ALKBH5 genes were closely related to E.coli resistance of piglets at the cellular and individual levels,which will provide a theoretical basis for future in-depth study of the regulation mechanism of RNA demethylation modification on bacterial diarrhea in piglets.  相似文献   

9.
为了探讨猪6-甲基腺嘌呤(m6A)去甲基化酶mRNA表达水平与仔猪大肠杆菌F18抗性的关系,本研究运用实时荧光定量PCR方法检测m6A去甲基化酶FTOALKBH5基因在35日龄苏太猪断奶仔猪大肠杆菌F18抗性型和敏感型个体十二指肠和空肠组织中的mRNA表达差异,同时分别利用F18ab、F18ac大肠杆菌菌体刺激和内毒素LPS诱导猪小肠上皮细胞(IPEC-J2),检测FTO、ALKBH5基因表达变化。结果表明,FTO、ALKBH5基因在抗性型个体十二指肠和空肠中的表达量均极显著或显著高于敏感型个体(P<0.01,P<0.05);不同F18大肠杆菌菌体刺激IPEC-J2细胞后,FTO基因表达水平均无显著变化(P>0.05),而ALKBH5基因在F18ac刺激后表达量显著上调(P<0.05);LPS诱导4 h时,FTOALKBH5基因表达量均显著上调(P<0.05)。本研究在细胞和个体水平上初步验证并发现了m6A去甲基化酶FTO和ALKBH5基因表达水平与大肠杆菌感染仔猪密切相关,为今后深入研究RNA去甲基化修饰在仔猪细菌性腹泻调控中的作用机制提供了理论基础。  相似文献   

10.
为研究鸭大肠杆菌的致病性及相关生物特性,本试验从西昌市某规模化鸭场采集病料,通过传统分离培养及理化性质鉴定,分离得到27株鸭致病性大肠杆菌。对27株鸭致病性大肠杆菌进行血清型鉴定、药敏试验、相关毒力基因检测。血清型鉴定结果显示,优势血清型为O119、O86、O126、O142和O44,占分离株55.56%。血清型O119占27株鸭致病性大肠杆菌的40.74%,为该鸭场流行的致病血清型。对20种兽医临床常用药物的药敏试验结果显示,27株鸭致病性大肠杆菌均对阿米卡星、庆大霉素和多黏菌素B敏感,对头孢曲松等10种药物较敏感,对利福平等5种药物耐受。大肠杆菌相关毒力基因检测结果显示, iutAhlyFIssIroNompTfyuAirp 2、TshpapA基因携带率均为100.00%, fimC基因携带率59.26%, K 99基因携带率7.40%。各项研究结果为有效防控鸭大肠杆菌病提供了重要科学依据,并为大肠杆菌深入研究奠定了基础。  相似文献   

11.
To analyze the pathogenicity and biological characteristics of duck E.coli, 27 strains of duck pathogenic E.coli were isolated from dead ducks with the typical characterize of colibacillosis by isolation and culture, physical and chemical properties identification in Xichang city.O serotype identification, drug sensitivity test and virulence associated genes were detected in 27 strains of duck pathogenic E.coli.The result showed that the predominant serotypes were O119, O86, O126, O142 and O44, which accounted for 55.56%.O119 was the epidemic and pathogenic serotype in this farm, which accounted for 40.74% in 27 strains of duck pathogenic E.coli.Through the drug sensitivity test of 20 kinds of clinical commonly used drugs, we found that all strains were sensitive to amikacin, gentamicin and polymyxin B.Besides, 10 drugs(ceftriaxone, etc) were lightly sensitive.However, these strains were resistant to 5 kinds of drugs(rifampicin, etc).The detection result of PCR about virulence associated genes indicated that the positive of iutA, hlyF, Iss, IroN, ompT, fyuA, irp 2, Tsh and papA genes were 100.00%, fimC and K 99 genes were 59.26% and 7.40%, respectively.The results provided an important reference for effective prevention and control of duck colibacillosis, and laid the foundation for further study of E.coli.  相似文献   

12.
The probiotics tested in the experiment were isolated from the intestinal of weaning piglets.The isolated probiotics and E.coli K88 were inoculated into the culture of intestinal porcine epithelial cell-1 (IPEC-1).The activity of lactate dehydrogenase (LDH) in the supernatant was measured after incubating for 2.5 h.At the same time, the probiotics and E.coli K88 were co-cultured in vitro, the number of E.coli K88 was counted and the probiotics which could be resistant to the E.coli K88 were selected 2.5 h later.The results showed that the Lactobacillus casei and Bacillus coagulans could significantly reduce LDH activity (P<0.05), decrease the damage of E.coli K88; Bacillus coagulans could inhibit the growth of E.coli K88.At the same time, Bacillus coagulans could resist high temperature, acid and bile salt.The results showed that Bacillus coagulans strains had great potential as the application of probiotics strains.The methods could be used as a model of screen probiotics which could inhibit the growth of E.coli K88 in vitro.  相似文献   

13.
本研究以从断奶仔猪肠道分离的益生菌为试验菌株,将大肠杆菌K88和益生菌接种到体外培养的猪小肠上皮细胞(intestinal porcine epithelial cell-1,IPEC-1)中,测定培养2.5h上清液中的乳酸脱氢酶(lactate dehydrogenase,LDH)活性,同时将益生菌和大肠杆菌K88体外混合培养2.5h,进行平板计数,统计大肠杆菌K88菌数的变化,筛选出可以抑制大肠杆菌K88的益生菌。试验结果显示,干酪乳杆菌和凝结芽孢杆菌能显著降低IPEC-1培养上清液中的LDH活性(P<0.05),降低大肠杆菌K88对细胞的损伤;凝结芽孢杆菌能降低DMEM培养基中大肠杆菌K88的生长速度,结合模拟制粒过程及胃肠道环境的耐高温、耐酸及耐胆盐研究进行综合分析,该凝结芽胞杆菌对大肠杆菌K88有较好的抑制作用且具有良好的耐高温、耐酸及耐胆盐性能,具有作为微生态制剂菌株的应用潜力。本试验建立了能够抑制大肠杆菌K88的益生菌体外筛选技术模型。  相似文献   

14.
Our objective was to generate hypotheses about associations between management, climate, and the presence of Escherichia coli O157 in feedlot–cattle water tanks and in feedlot–cattle feed. Water samples from 710 tanks on 73 feedlots, and feed-samples from a subset of 504 pens on 54 feedlots, in four US states were tested for E. coli O157. Management and climate factors were ascertained by survey and observation. Escherichia coli O157 were isolated from 13% of the water tanks and at least one water tank was positive on 60% of the feedlots. The factors significantly associated with E. coli O157 in water were greater percentage of cattle shedding E. coli O157 in faeces within the same pen, higher concentration of total E. coli in the water, lack of the clarity of the water, the use of fly traps, the reported frequency of rodent sightings in the pen or alley area, and the weather at the time of sampling. Escherichia coli O157 were isolated from 14.9% of the feed samples obtained from the feedbunks. Factors positively associated with E. coli O157 in feed were higher heat index at the time of sampling, the presence of cottonseed meal in the ration, and the feedlot location (state). Coliform counts in feed, presence of E. coli O157 in water tanks and faecal prevalence of E. coli O157 were not associated with the presence of E. coli O157 in feed.  相似文献   

15.
冯世文  李军  曾芸  杨威  陈泽祥  潘艳  彭昊 《中国畜牧兽医》2015,42(12):3315-3322
为初步研究猪源大肠杆菌O157:H7 (E.coli O157:H7)对氟苯尼考耐药性的产生和消除机制,本研究采用亚抑菌浓度体外耐药诱导的方法将两株猪源大肠杆菌O157:H7诱导成氟苯尼考高度耐药菌株,采用无氟苯尼考压力下连续传代培养的方法将获得的氟苯尼考耐药菌株的氟苯尼考耐药性消除,检测耐药诱导菌和耐药消除菌对抗菌药物的敏感性,并检测菌株质粒携带的耐药基因。结果显示,经氟苯尼考耐药诱导,猪源大肠杆菌O157:H7对氟苯尼考、阿莫西林、头孢唑啉、头孢拉定和头孢噻吩由敏感变为耐药,对头孢噻肟的敏感性由敏感变为中介,对氧氟沙星、环丙沙星和阿奇霉素由中介变为耐药;而经耐药消除后,菌株恢复对上述药物的敏感性;在菌株的质粒中检测到氟苯尼考耐药基因、喹诺酮类耐药基因和β-内酰胺酶基因,与耐药表型相符。结果表明,在氟苯尼考压力的长期存在下,猪源大肠杆菌O157:H7对氟苯尼考产生耐药,且对青霉素类、头孢类和喹诺酮类药物产生交叉耐药,在去除氟苯尼考压力下连续培养,可消除菌株的部分耐药性。  相似文献   

16.
The local and systemic immune response to a formolized E. coli oral vaccine was investigated in 13 gnotobiotic piglets. Beginning at ten days of age animals received a daily dose of 1010 or 1011 bacteria, on ten consecutive days. Intestinal loop tests with one animal of each group on day 26 showed protection which was more pronounced in the animal dosed 1010 bacteria compared with the other immunized piglet. Immunoglobulin class-specific antibodies to O and K antigens were determined by ELISA technique. In serum no IgG or IgA antibodies were found, whereas IgM-anti O149 antibodies in both immunized groups reached their highest level at day 4 of dosing and decreased thereafter. IgM-anti K88 antibodies were first detected at day 10 of dosing. Both immunized groups had comparable serum levels at days 20 and 30. Also in gut secretion the IgM antibody response was predominant, and higher levels were found in the 1010 group than in the 1011 group. IgG and IgA antibody response were also detected in secretion.  相似文献   

17.
本研究旨在探讨猪m6A甲基化酶WTAP表达水平与大肠杆菌(E. coli)感染抗性的关系。选取35日龄苏太断奶仔猪(Sus scrofa)大肠杆菌抗性型和敏感型个体各4头,采集十二指肠和空肠组织,利用RT-qPCR检测WTAPE. coli抗性型和敏感型个体十二指肠、空肠的表达差异,并分别利用产肠毒素大肠杆菌(F18ab、F18ac)刺激和内毒素(LPS)诱导猪小肠上皮细胞(IPEC-J2),检测WTAP基因的表达变化。同时构建WTAP基因干扰载体并转染IPEC-J2细胞,通过菌毛定量、菌落计数以及间接免疫荧光试验检测该基因沉默对大肠杆菌黏附能力的影响。结果显示:在十二指肠和空肠组织中,WTAP基因在E. coli抗性型个体中的表达量显著高于敏感型个体(P<0.01);并且在F18ab和F18ac刺激后表达量显著下降,与LPS诱导6 h后结果相一致(P<0.01)。沉默WTAP基因后,大肠杆菌黏附能力极显著上升(P<0.01)。本研究在细胞和个体水平上验证发现,m6A甲基转移酶WTAP的高表达可能有助于仔猪抗大肠杆菌感染,为进一步揭示仔猪抗大肠杆菌感染的RNA甲基化调控机制奠定基础。  相似文献   

18.
为建立牛源大肠杆菌-秀丽隐杆线虫致病模型,本研究从广西南宁、桂林、柳州等地区收集的牛病料中分离出13株大肠杆菌,继而对这些大肠杆菌进行血清学鉴定及小鼠、秀丽隐杆线虫的致病性试验。采用玻片凝集法鉴定大肠杆菌的O血清型,并将13株大肠杆菌培养液以3.0×109 CFU/mL、0.2 mL/10 g体重给小鼠腹腔注射,同时分别喂食N2野生型秀丽隐杆线虫。结果显示,大肠杆菌的O血清型为O127、O126和O44,其中O126为优势血清型(4/13)。致病性结果显示,有11株大肠杆菌能使小鼠致死(致死率为40%~100%),2株大肠杆菌对小鼠没有致死性(致死率为0)。对小鼠有强致病性的大肠杆菌,对线虫的致死率也较高,死亡率在第3~6天最为显著,半数致死时间为3~4.5 d,最长存活时间为9 d;对小鼠不致死的两株大肠杆菌对线虫的致死率也较低,线虫死亡率下降趋势缓慢,半数致死时间为5~6 d,最长存活时间为10~11 d。肠道细菌计数结果显示,大肠杆菌在线虫体内的数量与时间呈线性关系,大肠杆菌不断破坏线虫的免疫系统,从而导致了线虫的死亡。本研究结果表明,大肠杆菌对线虫和小鼠的致病力试验结果一致,说明成功建立了牛源大肠杆菌-秀丽隐杆线虫致病模型,为牛病防治与临床用药提供了依据。  相似文献   

19.
采用大肠杆菌C83549(O149:K88ac)、C83644(O68:K99)、C83710(O9:987P)菌株制备抗原,以蜂胶为佐剂,研制仔猪大肠杆菌病三价蜂胶灭活疫苗,疫苗成品检验均符合相关要求。将疫苗免疫妊娠母猪并用凝集试验测定其所产仔猪的抗体消长规律,并研究疫苗免疫保护相关性。结果表明:妊娠母猪产前40d和15d各免疫1次所产仔猪母源抗体效价较高,仔猪在整个哺乳期都可以得到保护。同时也可运用产前30d和10d进行免疫接种方法。  相似文献   

20.
旨在了解陕西省部分地区腹泻羊源致病性大肠杆菌(E. coli)耐药性及毒力基因携带情况,本研究从10个养殖场采集54份腹泻羊拭子样品,经分离纯化、生化鉴定及16S rRNA基因序列分析,共分离得到50株E. coli,对分离菌进行药敏试验、耐药基因及毒力基因检测。结果显示,分离菌对氨苄西林、氟苯尼考和磺胺异噁唑耐药率达90%以上,且98%(49/50)为多重耐药菌,对8~11种抗生素耐药的菌株占68%(34/50),仅对美罗培南敏感。所有菌株均携带1~6种不同的耐药基因,其中,Sul1(64%)、TetA(34%)、blaCTX-M(32%)携带率较高,未检测到blaSHV。有5株产ESBLs的E. coli携带mcr-1耐药基因。毒力基因检测结果显示,98%(49/50)的菌株携带毒力基因,其中,etrA检出率最高,为80%(40/50)。综上表明,陕西省羊源E. coli多重耐药情况严峻,β-内酰胺类耐药基因与耐药表型不符,提示可能存在其他耐药机制,同时,分离菌具有复杂的毒力谱。本研究为陕西省羊源致病性E. coli感染的防控提供科学依据。  相似文献   

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