Serological comparison of the Escherichia coli prototype strains for the F(Y) and Att 25 adhesions implicated in neonatal diarrhoea in calves

Slide agglutination tests using single absorbed and double absorbed antisera indicated that the Att 25 prototype Escherichia coli strain 25 KH9 produces the F(Y) adhesion; that this E coli also produces at least one other surface antigen not found on the F(Y) prototype E coli strain 11a; and that F(Y)+ E coli strain 28a produces at least one other surface antigen not produced by the prototype strains for the F(Y) and Att 25 antigens. These antigens were found on E coli isolated from outbreaks of calf diarrhoea in the United Kingdom.     

产志贺毒素样大肠杆菌F18ab菌毛操纵子基因的体外非诱导系统的表达和鉴定

以产志贺毒素样大肠杆菌（SLTEC）F18ab血清型标准菌株107/86基因组DNA为模板,利用PCR技术成功扩增出编码F18ab完整菌毛操纵子fed基因,克隆入表达载体pBR322,经限制性内切酶酶切分析,DNA琼脂糖电泳鉴定并结合序列测定分析,构建和筛选出含fed完整基因正确插入的pBR322-fed重组质粒,将上述重组质粒转化至不含任何菌毛结构的大肠杆菌SE5000,该表达重组菌能分别与兔抗F18ab亚单位蛋白FedF高免血清、鼠抗F18ab菌毛a单因子单克隆抗体、兔抗F18ab菌毛高免血清和抗F18ab菌毛IgG抗体产生明显的凝集反应。用热抽提法分别抽提和纯化SLTEC F18ab标准株107/86和重组菌SE5000（pBR322-fed）体外表达的F18ab菌毛,纯化菌毛经SDS-PAGE电泳和考马斯亮蓝染色获单一相对分子质量约为15 000蛋白条带。Western-blotting结果表明：兔抗F18ab菌毛高免血清能特异性识别SLTEC F18ab标准株107/86和重组菌SE5000（pBR322-fed）所提纯的单一主要结构蛋白。用重组菌SE5000（pBR322-fed）进行易感仔猪小肠上皮细胞体外黏附试验和黏附抑制试验,结果表明：重组菌SE5000（pBR322-fed）和SLTEC F18ab标准株107/86一样具有较强的黏附易感仔猪小肠上皮细胞的能力,而兔抗F18ab菌毛高免血清能有效地抑制上述重组菌SE5000（pBR322-fed）和SLTEC F18ab标准株107/86对易感仔猪小肠上皮细胞的黏附结合。     

The use of polymerase chain reaction for determination of virulence factors of Escherichia coli strains isolated from pigs in Poland.

E. coli strains isolated from pigs with postweaning diarrhea or edema disease were tested by phenotypic and genotypic methods for the presence of virulence antigens and genes, respectively. The slide agglutination and ELISA analyses were used for determination of F4, F5, F6, F17, and F41 fimbriae whereas the prevalence of fimbrial fedA and toxin eltI, estI, estII, stx1, stx2 and stx2e genes were recorded by the means of PCR. Only F4 antigen (ac variant) was found in strains of the serogroup O149:K91 isolated from pigs with diarrhea. PCR analyses showed that the fedA gene encoding F18 fimbriae was present in 61.9% of strains isolated from pigs with diarrhea and in 84.2% of strains isolated from pigs with edema disease. The eltI genes encoding heat-labile toxin I (LTI) were present only in 9 out of 21 strains recovered from pigs with diarrhea. Shiga toxin 2 variant (stx2e) genes were found in six isolates from edema disease and also in one strain from diarrhea. The PCR test used in the study was a sensitive and valuable method for determination of virulence factors of E. coli strains.     

AFA and F17 adhesins produced by pathogenic Escherichia coli strains in domestic animals.

AFA and F17 are afimbrial and fimbrial adhesins, respectively, produced by pathogenic Escherichia coli strains in domestic animals. F17-related fimbriae are mainly detected on bovine and ovine E. coli associated with diarrhoea or septicaemia. The F17-G adhesin subunits recognize N-acetyl-D-glucosamine (GlcNAc) receptors present on bovine intestinal cells. Some F17 subtypes also bind to GlcNAc receptors present on human uroepithelial and intestinal Caco-2 cells or to the laminin contained in the basement of mammalian membranes. F17 is often associated with other virulence factors (aerobactin, serum resistance, CNF2 toxin, K99, CS31A or AFA adhesins) on pathogenic E. coli. A cluster of only four genes is required to synthesize functional F17-related fimbrial structures. The hypothesis of multifunctional F17 fimbrial subunits is supported by the fact that: i) the N-terminal part of the adhesin subunit participates in receptor recognition, whereas the C-terminal part is required for biogenesis of the fimbrial filament; and ii) the interaction between structural and adhesin subunits seems to be crucial for the initiation of monomer polymerization. Recently, determinants related to the afa gene clusters from human pathogenic E. coli associated with intestinal and extra-intestinal infections were identified in strains isolated from calves and piglets with diarrhoea and septicaemia. Two afa-related gene clusters, designated afa-7 and afa-8, that encode afimbrial adhesins were cloned and characterized from bovine pathogenic E. coli. These animal afa gene clusters were plasmid and chromosome borne and were expressed by strains that produced other virulence factors such as CNF toxins, F17, PAP and CS31A adhesins. A high frequency of afa-8 and a low prevalence of afa-7 among bovine E. coli isolates were suggested by preliminary epidemiological studies. As with the human afa gene clusters, the animal ones encode an adhesive structure composed of two proteins: AfaE which mediates adhesion to epithelial cells and AfaD which is an invasin.     

Bacterial adherence in mastitis caused by Escherichia coli.

The possible role of bacterial adherence in the pathogenesis of experimental mastitis in the mouse was examined with four strains of Escherichia coli. Two of these strains had a known adhesion antigen (K88) and two did not. The K88 antigen did not play a significant role in the virulence or infectivity of E. coli either in the murine or bovine mammary gland. Two E. coli strains, W1 (K88+) and J2 (K88-) were virulent in the mouse but did not adhere to epithelial cells. Both these strains produced clinical mastitis in the cow. A third strain, D282 (K88-), produced mild disease in the mouse but was avirulent in the cow. The fourth strain, 233/ID (K88+), was avirulent in both the mouse and the cow. Strains D282 and 233/1D were killed rapidly by bovine serum whilst J2 and W1 were more resistant. All strains were more sensitive than the control resistant strain E. coli P4, which is known to be highly virulent for the lactating udder.     

Occurrence and characteristics of CS31A antigen-producing Escherichia coli in calves with diarrhoea and septicaemia in Argentina

CS31A is a K88-related non-fimbrial adhesin first described on Escherichia coli strains isolated from diarrhoeic and septicaemic calves. In this report, CS31A antigen was screened by immunological methods and confirmed by PCR among bovine E. coli isolates. In addition, CS31A-producing strains were characterized with respect to different fimbrial antigens, O-serogroup and other properties related to virulence. Faecal or tissue specimens of 100 diarrhoeic or septicaemic calves and 27 older cattle with different pathologies from 71 outbreaks or individual cases that occurred in Buenos Aires province, Argentina, were examined. CS31A + E. coli strains were isolated from 21 (21.0%) calves from 16 outbreaks or individual cases. No CS31A + E. coli was detected in samples from cattle more than 1 year old. Fimbriae F5, F41, F17a and F17b were not detected among the CS31A-producing strains. Three (14.3%) of the CS31A+ E. coli strains expressed the F17c fimbria. All of the 21 isolates exhibited at least one property of septicaemic strains (resistance to serum, production of aerobactin or colicins) but none of them demonstrated heat-stable enterotoxigenic activity. CS31A + E. coli isolates belonged to 10 serogroups, more commonly O8, O7, O17 and O21. The results obtained here confirm the worldwide distribution of CS31A antigen in bovine E. coli strains. However, CS31A + or CS31A + /F17c + E. coli were less frequently isolated than they were in North hemisphere countries.     

In vitro adhesion of an avian pathogenic Escherichia coli O78 strain to surfaces of the chicken intestinal tract and to ileal mucus

The role of fimbria in adherence of an avian pathogenic Escherichia coli (APEC) O78 strain 789 to chicken intestine was studied. Bacterial adhesion to tissue sections representing the regions within the chicken intestinal tract was determined by using immunohistochemical methods. E. coli 789 grown to express the type 1 fimbria adhered efficiently to the crop epithelium, to the lamina propria of intestinal villi, and to the apical surfaces of both the mature as well as the crypt-located enterocytes in intestinal villi, whereas no adhesion to mucus-producing goblet cells was detected. The adhesion was inhibited by mannoside and the role of type 1 fimbriae in the observed adhesion was confirmed with a recombinant strain expressing type 1 fimbriae genes cloned from E. coli and Salmonella enterica. E. coli 789 strain grown to favor AC/I fimbriae expression as well as the recombinant E. coli strain expressing the fac genes adhered to goblet cells but only poorly to the other epithelial sites. E. coli strain 789 as well as S. enterica serovar Typhimurium IR715 and S. enterica serovar Enteriditis TN2 strains were able to multiply in ileal mucus medium. The type 1 fimbria expressing bacteria adhered to the ileal mucus, whereas the AC/I fimbriated strains showed poor adherence to the mucus. The adhesion of E. coli 789 onto the crop epithelium and the follicle associated epithelium of the chicken ileum was efficiently inhibited by an adhesive strain ST1 of Lactobacillus crispatus isolated from chicken, whereas poor inhibition of E. coli adherence was observed with the weakly adhesive L. crispatus strain 134mi. The type 1 fimbriae may be important in colonization of the chicken intestine by APEC and Salmonella.     

Effect of antibodies to type 1 fimbriae on clearance of fimbriated Escherichia coli from the bloodstream of turkeys

Young turkeys (n = 20) were inoculated IV with fimbriated, virulent Escherichia coli ECl (O78:K80: H9:F1). Blood samples were collected for bacterial quantitation at postinoculation minutes (PIM) 10, 20, 30, 40, 50, and 60. Immediately after the PIM 30 sampling, the turkeys were allotted into 4 groups (5 turkeys/group) and were injected IV with 1 of the following antisera: group 1, antibodies to F1 fimbriae (AF); group 2, antibodies to E coli O78 antigen (AO); group 3, antibodies to live, fimbriated (F1+) homologous E coli (ALEC); or group 4, normal turkey serum (NTS) collected from a healthy turkey. Compared with NTS, ALEC and AO caused a significant reduction in blood-borne E coli, whereas AF did not reduce bacterial numbers. In addition, 2 groups of 10 turkeys were inoculated IV with live, F1+ or nonfimbriated (F1-) E coli ECl. Numbers of viable bacteria were determined in blood samples and liver specimens collected 2 minutes after inoculation. Compared with F1- bacteria, significantly more F1+ bacteria were found in liver specimens and significantly fewer F1+ bacteria were found in blood samples. Results indicated that antibodies to F1 fimbriae do not enhance clearance of F1+ E coli from the bloodstream of turkeys probably because F1+ bacteria are selectively cleared by the liver, even without antibody.     

Development of a sandwich ELISA and comparison with PCR for the detection of F11 and F165 fimbriated Escherichia coli isolates from septicaemic disease in farm animals

The P fimbriae F11 and F165 that have been demonstrated on Escherichia coli septicaemic strains in poultry and calves, respectively, possess a nearly identical major subunit that demonstrates a serological cross-reaction. A polyclonal antibody-based sandwich ELISA (sELISA) that was specific for both F11 and F165 fimbriated strains was compared with a PCR method to detect F11/F165 fimbriated strains, in a collection of E. coli strains isolated from diseased animals. Of 298 isolates tested, 36 were positive by PCR of which only 14 were sELISA positive. There were no sELISA positive but PCR negative results. The 36 PCR positive isolates comprised 11 avian strains of which 10 were sELISA positive, 20 bovine strains of which 4 were sELISA positive and 3 ovine strains, 1 porcine strain and 1 equine strain all of which were sELISA negative. The F11/F165 incidence of 10.7% in 103 poultry and 18.3% in 109 bovine isolates demonstrates a moderate level of these factors in E. coli septicaemic cases in Northern Ireland.     

Importance of Escherichia coli in young beef calves from northwestern Quebec.

The objectives of this study were: (i) to investigate the prevalence of Escherichia coli producing F5 (K99), F41, or F165 fimbriae and STa enterotoxin; (ii) to determine serum antibody levels against these fimbriae; (iii) and to examine the association between bacteriological and serological results and the presence of diarrhea, in beef calves from northwestern Quebec. A total of 373 live three to four week old calves and 27 dead calves were sampled between January and March 1991. No isolates positive for F5 were detected in live calves, and only one E. coli producing STa and F41 was isolated. Escherichia coli producing F41-like surface antigens or F165 fimbriae were isolated from 17.43% and 5.63% of live calves, respectively. Antibodies against F5, F41 and F165 were low. Escherichia coli isolates positive for F41-like surface antigen were most often observed in calves born between January and March. No association was found between bacteriological and serological findings, nor between these findings and diarrhea. Calves born from dams vaccinated against E. coli had higher median antibody levels than those born from unvaccinated dams. No E. coli positive for F5 or F41 fimbriae were isolated from dead calves. Escherichia coli with F41-like surface antigen or F165 were found in 55.56% and 11.11% of ileal samples; 4% and 16% of cecal samples, and 0% and 7.4% of colon samples, respectively. Escherichia coli positive for F41-like surface antigen were detected significantly more frequently in the ileum (chi (2)2df = 31.01, p < 0.001).     

鸡致病性大肠杆菌菌毛分型的初步研究

以鸡致病性大肠杆菌F1菌毛特异性单抗1F4－1、2C3、3H3和F11菌毛特异性单抗FA1、FB11作为检测试剂，将111株已知O抗原的鸡致病性大肠杆菌经MD液体培养基连续传代培养后，通过直接玻板凝集法对各菌毛进行初步分型。结果发现F1、F11菌毛与O78、O1及O2三种优势O抗原型菌株之间存在较为明显的相关性，即致病性大肠杆菌主要集中在上。F1、F11菌毛在这三种O抗原型上的总检出率分别为95．6％、75．4％及73．3％。另外，在所检测的111株鸡致病性大肠杆菌中，只表达F1菌毛的大肠杆菌占菌杆总数的33．3％。只表达F11菌毛的大肠杆菌占菌株总数的8．1％，两者都表达的占菌株总数的36％，F1、F11菌毛的总检出率为78．3％。     

卵黄抗体对ETEC粘附仔猪小肠上皮细胞的体外抑制作用研究

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。分别制成单价或多价的菌毛蛋白白油佐剂抗原 ,对产蛋鸡进行胸部肌肉分点注射免疫 ,初免后 2周加强免疫 1次。收集高效价卵黄抗体。用所获得各卵黄抗体对体外分离的初生仔猪小肠上皮细胞进行体外粘附抑制试验。结果表明 ,各种菌毛卵黄抗体均能特异地显著抑制相应大肠杆菌对仔猪上皮细胞的粘附 ,而对其他血清型大肠杆菌对肠上皮细胞的粘附无抑制作用     

Avirulent K88 (F4)+ Escherichia coli strains constructed to express modified enterotoxins protect young piglets from challenge with a virulent enterotoxigenic Escherichia coli strain that expresses the same adhesion and enterotoxins

Virulence of enterotoxigenic Escherichia coli (ETEC) is associated with fimbrial adhesins and enterotoxins such as heat-labile (LT) and/or heat-stable (ST) enterotoxins. Previous studies using a cell culture model suggest that exclusion of ETEC from attachment to epithelial cells requires expression of both an adhesin such as K88 (F4) fimbriae, and LT. To test the ability of non-pathogenic E. coli constructs to exclude virulent ETEC sufficiently to prevent clinical disease, we utilized a piglet ETEC challenge model. Thirty-nine 5-day-old piglets were inoculated with a placebo (control), or with either of the three K88(+)E. coli strains isogenic with regard to modified LT expression: 8017 (pBR322 plasmid vector control), non-toxigenic mutant 8221 (LT(R192G)) in pBR322, or 8488, with the LT gene fused to the STb gene in pBR322 (LT(R192G)-STb). Piglets were challenged with virulent ETEC Strain 3030-2 (K88(+)/LT/STb) 24h post-inoculation. K88ac receptor-positive piglets in the control group developed diarrhea and became dehydrated 12-24h post-challenge. Piglets inoculated with 8221 or 8488 did not exhibit clinical signs of ETEC disease; most piglets inoculated with 8017 showed diarrhea. Control pigs exhibited significant weight loss, increased blood total protein, and higher numbers of colony-forming units of 3030-2 E. coli in washed ileum and jejunum than treated pigs. This study shows for the first time that pre-inoculation with an avirulent strain expressing adhesive fimbriae and a non-toxic form of LT provides significant short term protection from challenge with a virulent ETEC strain that expresses the same fimbrial adhesion and enterotoxin.     

Characterization of a new fimbrial antigen present in Escherichia coli strains isolated from calves.

Thirteen Escherichia coli strains isolated from calves with diarrhoea, supposed to carry a common antigen were examined for their hemagglutinating activity and compared by bacterial agglutination, double diffusion in two dimensions and by crossed immunoelectrophoresis (CIE). Two of the strains were examined also in the electron microscope. Most of the strains agglutinated red blood cells of horse, ox, guinea pig and chicken, of which the agglutination of ox erythrocytes was mannose-resistant (MRHA). None of the strains agglutinated human erythrocytes. All strains with MRHA of ox red blood cells, regardless to their O:K:H antigens could be agglutinated in unabsorbed or absorbed antisera produced against cultures C1209 (020:K-:H9) and C1213 (09:K36:H-) when live cells as antigens were used. None of these sera agglutinated reference strains carrying K88, K99, 987P, F41 or FY (Att25) antigen respectively. By the double gel diffusion test and by CIE in extracts (60 degrees C) of the strains a common heat labile antigen, responsible for the MRHA of ox red blood cells was identified. Electron microscopy revealed that this common antigen was represented by thin, long, hair-like fimbriae on cells of E. coli C1213, and that specific homologous antibodies attached to these fimbriae.     

Identification and detection of three new F17 fimbrial variants in Escherichia coli strains isolated from cattle

F17 fimbriae are produced by pathogenic Escherichia coli involved in diarrhea and septicemia outbreaks in calves and lambs. These proteins result from the expression of four different clustered genes, namely f17A, f17D, f17C and f17G, encoding a pilin protein, a periplasmic protein, an anchor protein and an adhesin protein, respectively. Several variants of f17A and f17G genes have been reported and found genetically associated with typical virulence factors of bovine pathogenic E. coli strains. In this study, a new F17e-A variant, closely related to F17b-A, was identified from a collection of 58 E. coli isolates from diarrheic calves in Iran. While highly prevalent in Iranian F17-producing clinical isolates from calves, this variant was rare among E. coli from a French healthy adult bovine population, suggesting a possible association with virulence. The f17Ae gene was also found in the genome of the Shiga-like toxin variant Stx_1d–producing bovine E. coli strain MHI813, and belonged to a gene cluster also encoding a new F17-G3 variant, which greatly differed from F17-G1 and F17-G2. This gene cluster was located on a pathogenicity island integrated in the tRNA pheV gene. The gene coding for a third new F17f-A variant corresponding to a combination of F17c-A and F17d-A was also identified on the pVir68 plasmid in the bovine pathogenic E. coli strain 6.0900. In conclusion, we identified three new F17-A and F17-G variants in cattle E. coli, which may also have significant impact on the development of new diagnostics and vaccination tools.     

苏太仔猪FUT1基因M307位点多态性与F18大肠杆菌抗病相关性的体外鉴定

采用PCR-RFLP方法检测了江苏苏太断奶仔猪FUT1基因M307位点等位基因多态性分布,在所检的49头仔猪中,GG基因型个体16头,AG基因型19头,AA基因型14头。在此基础上,制备上述不同基因型个体仔猪小肠上皮细胞,分别与表达F18ab菌毛的野生型大肠杆菌、表达F18ac菌毛含fed操纵子全基因的重组大肠杆菌和V型系统表面分泌表达F18abFedF亚单位的重组大肠杆菌进行体外黏附试验和黏附抑制试验。研究结果表明:FUT1基因M307位点中GG型和AG型仔猪小肠上皮细胞均能黏附上述3种大肠杆菌,而AA型个体小肠上皮细胞则不能黏附。将上述3种大肠杆菌分别与抗F18ab菌毛高免血清、F18ac菌毛高免血清及抗F18abFedF亚单位单因子血清作用后,则失去黏附仔猪肠上皮细胞能力。上述结果对苏太猪从体外试验上证明了FUT1基因M307位点多态性与断奶仔猪腹泻和水肿病存在着直接的相关性。     

Evaluation of a monoclonal antibody to the K99 fimbrial adhesin produced by Escherichia coli enterotoxigenic for calves, lambs and piglets

All the K99+ Escherichia coli grown at 37 degrees C stained strongly with a peroxidase labelled K99 monoclonal antibody using a direct immunoperoxidase staining procedure. There was no reaction when these bacteria were cultured at 18 degrees C or when K99- E coli were grown at either temperature. The binding of the monoclonal antibody to K99 antigen was inhibited by OK antisera to heterologous K99+ E coli but OK antisera to E coli producing adhesins other than K99 were without effect. Using the slide agglutination test the reactions of the monoclonal antibody were identical to those of a polyclonal antiserum to K99 when both were used in parallel to examine 100 K99+ E coli from at least 10 somatic O groups and 1308 K99+ E coli from at least 82 different somatic O groups submitted for routine serological typing in England or the, USA. The monoclonal antibody reacted with K99+ E coli in cryostat sections of the ileum from a piglet infected with E coli strain B44 (O9: K30, K99, F41) but there was no reaction with similar material from piglets infected by E coli strains 1751 (O101: F41), X177/81 (O9: K103, 987P) or Abbotstown (O149: K91, K88ac).     

Active oral immunization of suckling piglets to prevent colonization after weaning by enterotoxigenic Escherichia coli with fimbriae F18

Immunoprophylaxis of porcine oedema disease and post-weaning diarrhoea caused by strains of Escherichia coli expressing fimbriae F18 is an unsolved problem. The study was designed to examine whether vaccination with a live F18ac vaccine of unweaned pigs born to sows with F18ac antibody in the colostrum requires preformed fimbriae in the vaccine, and whether protection against the heterologous fimbrial variant F18ab is induced as well. Genetically susceptible pigs were vaccinated orally on three consecutive days, beginning 10 days before weaning with 10(11) CFU of an F18ac culture. Challenge with a dose of 10(7) CFU of E. coli F18 on three consecutive days was initiated 9 or 11 days after weaning. Eighteen pigs given the fimbriated F18ac vaccine and challenged with a strain of the homologous fimbrial variant were protected against colonization; mean faecal viable counts of the challenge strain were >3 log10 lower than those from the 17 non-vaccinated control pigs. The vaccinated pigs developed a significant rise of F18ac IgA serum antibodies. The 23 pigs which had received the non-fimbriated vaccine showed no significant protection and exhibited much lower serum F18ac IgA ELISA reactivities. Eighteen pigs vaccinated with the fimbriated F18ac and challenged with an F18ab strain had faecal viable counts nearly as high as those from 16 non-vaccinated control pigs. It is concluded that only oral vaccines having preformed fimbriae induce protection limited to the homologous fimbrial variant.     

Characterization of Escherichia coli isolated from adult horses with and without enteritis

In the present study E. coli strains isolated from the faeces of ten horses with diarrhoea and 14 horses without diarrhoea were characterized. All horses were culture negative for Salmonella species. Nine colonies of E. coli from each faecal sample were picked at random and a DNA fingerprint was made by means of a polymerase chain reaction (PCR) using Enterobacterial Repetitive Intergenic Consensus (ERIC) primers. The number of E. coli genotypes did not differ significantly between horses with and without diarrhoea. In addition, all E. coli strains with different DNA fingerprints were tested by PCR for genes encoding the virulence factors K88, F41, F17, CS31a, Sta1, LT1, VT2, CNF, BFP, and intimin. Genes coding for K88, F41, BFP, STa1, VT2, and CS31A were not detected. Genes for CNF were found in strains from one horse with diarrhoea and one horse with normal faeces. Genes for LT1 (n=1) and intimin (n=1) were found only in strains from horses with normal faeces. Genes for F17 fimbriae were found in strains from three horses with diarrhoea (30%) and in none of the strains from healthy horses. In two of these horses, E. coli strains with different DNA polymorphism patterns were F17 positive; however, none of these strains possessed LT1, Sta1, or CNF genes. Haemolytic E. coli strains were only isolated from two horses with diarrhoea and from none of the healthy horses. Nineteen percent of all E. coli strains did not ferment lactose. Eight per cent of these lactose-negative strains were from horses with diarrhoea, whereas 32% were from horses without diarrhoea. In conclusion, virulence factors were present in E. coli isolates from horses with and without diarrhoea, except for F17, which was only found in E. coli isolated from horses with diarrhoea. F17-positive E. coli might have importance as cause of diarrhoea in horses, but further studies are needed.     

Virulence factors and antimicrobial susceptibility of Escherichia coli isolated from calves in Turkey

Escherichia coli isolates from calves were investigated by multiplex PCR assays for the presence of genes encoding K99, F41, F17-related fimbriae, heat-stabile enterotoxin a (STa), intimin (eae) and Shiga toxins (stx1 and stx2). A total of 120 E. coli isolates, 75 isolated from diarrhoeic or septicemic calves and 45 from clinically healthy calves aged between 1 day and 2 months were tested. Each isolate was obtained from different calves in different herds. Among the isolates from diseased animals, 12 (16%) isolates from 1- to 7-day-old diarrhoeic calves were detected as enterotoxigenic E. coli which possessed K99, F41 and STa in combination; F17-related fimbriae genes were detected in 33 (44%) isolates and they were found in combination with K99 + F41 + STa in two isolates. Of 120 isolates, 16 carried eae, eight stx1 and five stx2 genes alone or in combination. None of the eae- or stx-positive strains was identified as O157:H7. However, results indicate that calves may be carrier of Shiga toxin-producing E. coli which have potential as a human pathogen. Antimicrobial susceptibility of 75 isolates from diseased calves was determined by agar disk diffusion method for 14 antimicrobial agents. In 77.3% of the isolates, multiresistance was detected. Higher resistance rates were detected for cephalothin (72%), tetracycline (69.3%), kanamycin (69.3%), ampicillin (65.3%), nalidixic acid (53.3%), trimethoprim-sulphamethoxazole (52%) and enrofloxacin (41.3%), respectively. No resistance was found for ceftiofur and cefoxitin.