Antimicrobial drug resistance in porcine enterotoxigenic Escherichia coli of 0-group 149 and non-enterotoxigenic Escherichia coli

Enterotoxigenic Escherichia coli (ETEC) strains (104) and 96 non-ETEC, all isolated from herds with piglet diarrhea in 1981 and 1982, were investigated with regard to antimicrobial drug resistance and colicinogeny. Eighty strains (77%) of the ETEC were drug resistant as compared to 66 strains (69%) of the non-ETEC. There were no significant differences between ETEC and non-ETEC in the frequencies of the drug resistance determinants investigated, except for tetracycline, to which 51 (49%) of the former and 30 (31%) of the latter strains were resistant (0.05 greater than P greater than 0.01). Of all strains 116 (58%) were resistant to streptomycin, 93 (47%) to sulphadimidin, 81 (41%) to tetracycline, 20 (10%) to trimethoprim, 14 (7%) to ampicillin, 11 (6%) to neomycin, 6 (3%) to chloramphenicol and none to nitrofurantoin. The frequencies of the different drug resistance determinants correlated well with the total amount of active substance of each drug used in farm animals in Sweden in 1981. Of the ETEC, 97 (93%) were colicinogenic whereas only 13 (14%) of the non-ETEC were colicinogenic.     

A homolog of the O157 urease-encoding O island 48 is present in porcine O149:H10 enterotoxigenic Escherichia coli

The relationship of the urease operon in the highly virulent O149 porcine enterotoxigenic Escherichia coli (ETEC) strain Ro8 to a genomic island (GI) homologous to O island (OI) 48 of O157 enterohemorrhagic E. coli (EHEC) strain EDL933 was investigated. Eighty-four of 84 O149:H10 strains were urease positive whereas 44 of 44 O149:H43 porcine ETEC strains were urease-negative. Seventeen of 17 O149:H10 strains that were tested possessed the OI-48 homolog whereas 24 of 24 O149:H43 strains lacked this OI. Transposon insertions in lipB or guaA genes in strain Ro8 eliminated urease activity while insertions in the caiF gene increased urease activity. When the O149 ure operon was cloned on a high copy number plasmid, urease expression was increased approximately 11-fold in Ro8 and 83-fold in O157 strain EDL933 compared with that in the wild type Ro8. The O149 urease activity was expressed despite the presence of the same premature stop codon in ureD that is present in ure+ O157:H7 strains that are urease-negative. The ure operon in Ro8 consists of 4 893 nucleotides with 99% identity with the ure operons in EHEC O157:H7 strains EDL933 and Sakai, and is part of a GI similar to GI-48 of strain EDL933. This OI, designated OI-48149 , is inserted in the serX tRNA gene in strain Ro8 and contains genes for urease, tellurite resistance, iha and an AIDA-I-like adhesin. The presence of a homolog of the O157:H7 OI-48 in highly virulent O149 porcine ETEC suggests that this OI may contribute to establishment of the bacteria in the intestine.     

Isolation of enterotoxigenic Escherichia coli from a foal with diarrhea

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.     

Isolation of enterotoxigenic Escherichia coli from camels with diarrhoea.

E. coli serogroups 02, 08, 083, 0103 and 0120 were isolated from seven camels with diarrhoea of which 02, 08, and 083 were found to be enterotoxigenic on rabbit ligated ileal loop test. Out of 125 apparently healthy camels, 75 strains of E. coli were isolated. The majority of isolates were susceptible to gentamycin, nitrofurantoin, trimethoprim plus sulphonamide, neomycin, kanamycin and chloramphenicol.     

Naturally occurring bacteriophages lyse a large proportion of canine and feline uropathogenic Escherichia coli isolates in vitro

We investigated the feasibility of bacteriophage therapy to combat canine and feline Escherichia coli urinary tract infections (UTIs) by testing the in vitro lytic ability of 40 naturally occurring bacteriophages on 53 uropathogenic E. coli (UPEC). The mean number of UPEC strains lysed by an individual bacteriophage was 21/53 (40%, range 17-72%). In total, 50/53 (94%) of the UPEC strains were killed by one or more of the bacteriophages. Ten bacteriophages lysed 51% of UPEC strains individually and 92% of UPEC strains as a group. Electron microscopy and DNA sequencing of 5 'promising' bacteriophages revealed that 4 bacteriophages belonged to the lytic T4-like genus, while one displayed morphologic similarity to temperate P2-like bacteriophages. Overall, these results indicate that the majority of UPEC are susceptible to lysis by naturally occurring bacteriophages. Thus, bacteriophages show promise as therapeutic agents for treatment of canine and feline E. coli UTIs.     

Characterization of enterotoxigenic bovine Escherichia coli.

Among 300 isolates of bovine Escherichia coli, 56 which had been found enterotoxigenic in calf gut loops were characterized on the basis of O and K antigens, colonial morphology and resistance to seven antimicrobial drugs. The 56 isolates enterotoxigenic in the calf were compared with the nonenterotoxigenic ones. Of the 56 enterotoxigenic E. coli the majority possessed the A type of K antigen and had OK groups, O9:K(PS274) or O101:K(RVC118). Fourteen of these isolates had the K99 antigen. None of 27 isolates found enterotoxigenic in the piglet but not in the calf possessed the K99 antigen or belonged to OK groups O9:K(PS274) or O101:K(RVC118). Comparison of the patterns of resistance to seven antimicrobial drugs showed that all enterotoxigenic and nonenterotoxigenic isolates were susceptible to nitrofurantoin and sulphachlorphyridiazine and that there was no significant difference in the patterns between the two groups. The majority of enterotoxigenic isolates were mucoid, whereas most of the nonenterotoxigenic isolates were nonmucoid.     

产肠毒素大肠杆菌感染的分子致病机制

产肠毒素大肠杆菌（enterotoxigenic Escherichia coli,ETEC）是发展中国家人群腹泻的主要原因,一直是西方国家旅行者腹泻的最常见病因,也是引起动物（尤其是幼龄动物）腹泻的主要病原菌。根据ETEC产生热敏和（或）热稳定肠毒素特性可以将其分类成不同致病型。针对这类重要病原体的疫苗研发目前仍然面临着艰难的挑战。本文综述了病原体-宿主相互作用的分子机制,从基因组学和蛋白质组学两方面介绍致病菌的多种毒力因子以及作为靶标研发疫苗的潜力,分析了病原与不同宿主易感性的差异,为针对ETEC新型疫苗的研发和有效预防ETEC感染提供理论依据。     

Isolation and characterization of type 1 fimbriae from a chicken pathogenic Escherichia coli serotype O78.

Type 1 fimbriae from chicken pathogenic Escherichia coli strain PDI-386 (serotype O78) was purified and characterized. Because of the acid-induced autoagglutination (T. Sekizaki, Y. Nakasato, and I. Nonomura, J. Vet. Med. Sci. 54, 493-499, 1992), the fimbriae could be easily purified by repeating acid sedimentation, washing, and dissolving in buffer (pH 8.0). In electron microscopy, the purified fimbriae showed a filament of 8 nm in diameter and 10 microns in average length. The molecular mass of the protein subunit of the purified fimbriae estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was 19,000 daltons. The amino acid composition and its NH2-terminal sequence were similar to the previously described one of the Klebsiella pneumoniae type 1 fimbriae. Moreover, there was an immunological relatedness between them. These results indicated that a molecular diversity found between the fimbriae of E. coli and that of K. pneumoniae has already been existed among chicken pathogenic E. coli strains.     

Experimental infection with Escherichia coli O149:F4ac in weaned piglets

The outcome of experimental intestinal infections with enterotoxigenic Escherichia coli (ETEC) is dependent on several factors. An important factor is adhesion of the challenge strain to the intestinal mucosa. The test for susceptibility towards ETEC adhesion has so far been made by an intestinal adhesion test made after slaughter of piglets. However, in an experimental infection study with the purpose to obtain diarrhoeic piglets, it would be an advantage to test for susceptibility prior to experimentation. The Mucin 4 gene on porcine chromosome 13 has been proposed as a candidate gene for the production of the specific ETEC F4ab/ac receptor, and a DNA marker-based test has been developed to allow genotyping for ETEC F4ab/ac resistance/susceptibility [J?rgensen, C.B., Cirera, S., Archibald, A.L., Anderson, L., Fredholm, M., Edfors-Lilja, I., 2004. Porcine polymorphisms and methods for detecting them. International application published under the patent cooperation treaty (PCT). PCT/DK2003/000807 or WO2004/048606-A2]. The aim of this study was to test an experimental model for ETEC O149:F4ac-induced diarrhoea in piglets, selected for susceptibility towards ETEC O149:F4ac adhesion prior to experimentation using a DNA marker-based test. Sixty-two healthy 25-32 days old recently weaned Danish crossbred piglets were used. All piglets were tested prior to experimentation for susceptibility or resistance towards ETEC O149:F4ac adhesion. Thirty-nine piglets, both susceptible and resistant, were oro-gastric intubated with 10(9)CFU of ETEC O149:F4ac and 23 age-matched piglets, both susceptible and resistant, were used as non-infected controls. Of susceptible piglets, challenged with ETEC O149:F4ac, 74% had ETEC O149:F4ac-associated diarrhoea first day after first challenge, which were significantly higher relatively to the resistant and challenged piglets where 20% had diarrhoea (p=0.04). This study suggests a model for experimental ETEC induced diarrhoea.     

出血性大肠杆菌噬菌体933W的分离鉴定和保存

采用双层琼脂平板法从EHEC 0157 EDL933基因组中诱导分离出了编码志贺毒素的噬菌体933W，并利用PCR技术对其进行了鉴定。对933W的不同保存方法进行比较，结果表明，将噬菌体933W置于SM溶液中加入CaCl2、lgCl2至终浓度10mmol／L，明胶浓度至0．1％（W／V）并于4℃冷藏是保存933W较好的方法。     

Virulence plasmids of enterotoxigenic Escherichia coli isolates from piglets

Virulence plasmids of 68 ETEC isolates from piglets belonging to different pathotypes and six ETEC isolates from calves with pathotypes typical of porcine ETEC were identified with seven virulence probes for the heat-stable (STa and STb) and heat-labile (LT) enterotoxins, for the F4, F5, F6, and F41 fimbrial adhesin subunit, and also with five Rep probes for the RepFIA and RepFIB basic replicons, and the RepFIC family of basic replicons. With the exception of the F41 probe, the other virulence probes hybridized with at least one plasmid band of a size range from 65 to more than 100 Mda. Common associations of virulence factor-encoding genes on plasmid bands were: STb/LT, STa/F5, STa/F6, STa/STb. Other associations, STa/F4, STa/F4/F6, and STa/STb/LT/F6, were rarer. On the other hand the F4 adhesin-encoding genes were isolated on one plasmid band in all but three F4+ isolates. All but one of the 92 virulence plasmids which were studied have Rep probe hybridization profiles and replicon types typical of the uni- or multireplicon plasmids belonging to the various incompatibility groups of the F incompatibility complex.     

一株血清型O101多重耐药牛致病性大肠杆菌菌株的分离鉴定

试验对病料进行病原检测,并对分离到的病原菌进行分子生物学鉴定、血清学鉴定、药物敏感性试验、小鼠致病性试验研究,依据研究结果进行针对性治疗。试验结果:成功分离到一株牛致病性大肠杆菌,该株菌多重耐药,对小鼠具有高度致病性,其血清型为O101,使用该菌株的敏感药物对后续腹泻犊牛进行了及时治疗,成功控制了该养殖场犊牛腹泻的问题。     

Features of enterotoxigenic Escherichia coli strains of porcine origin that express K88 and 987P fimbrial antigens

Escherichia coli strains of porcine origin express K88 and 987P pilus-antigens in vitro. This study reports their enterotoxin producing ability, serological features and plasmid content. The bipiliated strains were enterotoxigenic and all contained a large plasmid of uniform size.     

Preliminary investigations of the distribution of Escherichia coli O149 in sows, piglets, and their environment

Little is known about the sources and kinetics of enterotoxigenic Escherichia coli colonization in pigs during the pre-and post-weaning period. In this study, farrowing pens, sows, and piglets were tested for the presence of E. coli O149 by real-time polymerase chain reaction (PCR) after bacterial culture pre-enrichment on 2 farms, one with a history of post-weaning diarrhea (problem farm - PF) and the other without such a history (non-problem farm - NPF). Unlike those on the PF, the sows from the NPF did not carry E. coli O149 before parturition, although they were colonized to frequencies similar to animals on the PF soon afterwards. Most piglets from the NPF were colonized within a week after birth, whereas only a small proportion of those on the PF were colonized during that period. No difference was observed in the frequency of piglet colonization at the 2 farms either at weaning or during the following week. Post-weaning diarrhea (PWD), which is caused by enterotoxigenic E. coli (ETEC), is a multifactorial disease. The presence of ETEC alone is not always sufficient for the disease to develop. Many other factors are considered to be associated with the occurrence of PWD, including feed type (1,2), feeding regimen (1,3,4), the presence of other infectious agents (3,5), weaning age, and weight (6). Weaning, which is considered to be a major physiological and psychological stress factor, is critical for the disease to occur (7). Although piglets are already colonized with ETEC before weaning (4,8), on many farms, clinical disease occurs only after weaning (1). Both sows (9,10) and the environment (6) could be possible sources of infection for piglets, but results from previous studies have not resolved this issue because of the low sensitivity of ETEC detection methods. This study provides preliminary data based on a sensitive detection method for E. coli O149 in pigs and their environment. The results demonstrate the potential of real-time PCR for future studies on this topic.     

猪源大肠杆菌O20的分离鉴定及其致病性研究

已知多种血清型的大肠杆菌能够引起仔猪发病，不同地区或同一地区的不同饲养场流行的大肠杆菌优势血清型不同，并且大肠杆菌耐药性普遍，给免疫预防和药物治疗造成困难。流行病学调查表明，大肠杆菌病是我国养猪场常见的有重要经济意义的细菌性传染病之一。因此，分析研究当地规模化猪场流行的大肠杆菌血清型、耐药性以及致病性，     

Isolation,O Serogrouping and Drug Sensitivity Test of Avian Pathogenic Escherichia coli

One hundred and ninety four avian pathogenic Escherichia.coli (APEC) isolated from different pathotypes of chickens in Hebei province,Shandong province,Henan province,Jiangsu province,Shanxi province and Liaoning province in China between May 2005 and March 2016 were investigated by determination of biochemical characters,O serogroup identification and drug sensitivity test.Out of 194 isolates, 124 strains(63.9%) could be grouped to serogroups O1(3),O2(26),O57(15),O65(47) and O78(33).The drug sensitivity test used eight antibiotics (ampicillin, neomycin, doxycycline, sulfamonomethoxine,tilmicosin,ciprofloxacin ceftiofur and florfenicol) by one strain of O1,O2,O57,O65,O78 serogroup,respectively.The results showed that all strains were sensitive to ciprofloxacin and neomycin, and resistance to other drugs at varying degrees.This study demonstrated that the high prevalence of O65,O78,O2 and O57 isolates existed and provided the basis for the development of multivalent bacterin in future.     

肠毒素性大肠杆菌菌毛对产蛋鸡免疫性的研究

采用热抽提法提取4种肠毒素性大肠杆菌菌毛蛋白。K88、K99、F41和987p菌毛蛋白分别制成弗氏佐剂苗;K88还制成白油佐剂苗,氢氧化铝胶苗和蜂胶佐剂苗;另将4种菌毛等比例混合制成弗氏佐剂苗。分别对产蛋鸡进行免疫,用微量凝集反应和血凝抑制试验检测卵黄抗体效价。结果表明,K88菌毛较其他3种菌毛免疫性好,诱导抗体效价最高而且能长时间维持;987p菌毛能快速诱导抗体的产生,但整体效价低。K88不同佐剂苗中,铝胶佐剂能较快地诱导抗体的产生,蜂胶佐剂苗抗体持续时间短,弗氏佐剂能诱导高效价的抗体产生而且能长时间持续。     

产肠毒素性大肠埃希氏菌基因工程疫苗的研究进展

简述了产肠毒性大肠埃希氏菌（ETEC）K88-K99、K88ac-LTB、K99-F41和ST-LT双价基因工程疫苗的研制方法、生产工艺、免疫效果及生态效应及其研究进展，同时阐明了用这些疫苗免疫妊娠母畜，可使吸吮了母畜初乳的幼畜的腹泻发病率和死亡率显著下降，且不会带来生态环境污染。认为这些基因工程疫苗仍有不足之处，需要进一步加以改进。     

禽致病性大肠杆菌的分离、O血清群鉴定及药物敏感试验

对2005年5月~2016年3月自河北、山东、河南、江苏、陕西、辽宁等省发病鸡场疑似大肠杆菌病的病死鸡病变部位分离的194株禽致病性大肠杆菌进行了生化特性测定、O抗原血清群鉴定及药敏试验。试验用5种大肠杆菌单因子（O1、O2、O57、O65、O78）阳性血清进行O血清群鉴定,确定了124株O血清群,占鉴定菌株数的63.9%（124/194）,其中O1 3株、O2 26株、O57 15株、O65 47株和O78 33株,O65和O78为优势菌株,而O2和O57血清群菌株次之,O1血清群菌株最少。8种药物（氨苄西林、新霉素、多西环素、磺胺间甲氧嘧啶、替米考星、环丙沙星、头孢噻呋和氟苯尼考）的药敏试验结果表明,受试的5株大肠杆菌分离株均对环丙沙星和新霉素敏感,而对其他药物均有不同程度的耐受。本研究结果表明O65、O78、O2和O57血清群高度流行,为进一步研发多价菌苗奠定了基础。