Attaching and effacing Escherichia coli infection as a cause of diarrhea in young calves

A form of enteric Escherichia coli infection was identified in 60 calves from 59 farming operations. The E coli responsible for these infections principally colonized the colon, inducing a distinctive lesion described as attaching and effacing. Hemorrhagic enterocolitis or blood in the feces was observed on 40% of the farms. Of affected calves, 86.6% were dairy calves (average age, 11.8 days). Forty-four calves were infected concurrently with other enteropathogens (cryptosporidia, rotavirus, coronavirus, enterotoxigenic E coli, bovine viral diarrhea virus, coccidia). Verotoxin-producing E coli was recovered from 31 calves; 8 were serotype O111:NM isolates, 3 were serotype O5:NM, and 1 was serotype O26:NM.     

The significance of bredavirus as a diarrhea agent in calf herds in Lower Saxony]

The objective of this investigation was to determine the distribution of Bredavirus in cattle herds in Lower Saxony and to evaluate its significance as potential cause of diarrhea in calves. Fecal samples and paired blood samples of 119 diarrheic and 46 healthy calves up to two months of age were collected from herds where diarrhea of calves was a problem. Fecal samples were examined for Breda-, rota- and coronavirus by solid phase immune electron microscopy and by ELISA, for K99-positive E. coli and salmonella by microbiological methods, and for cryptosporidia in smears. Antibody titers against Bredavirus, total serum protein and serum gamma globulin content were evaluated in the blood samples. Bredavirus was found in fecal samples from 5% (n = 6) of diarrheic calves which came from four different herds, but not in healthy calves. Rotavirus (31.9%), coronavirus (18.5%) and cryptosporidia (29.9%) were detected more frequently in fecal samples than Bredavirus. In this investigation rotavirus, coronavirus and cryptosporidia were present in addition in all herds where Bredavirus was found. In contrast to the low percentage of fecal samples containing Bredavirus, antibody titers in 75% of calves confirmed the high prevalence of Bredavirus infection in the cattle population of Lower Saxony.     

Enzyme-linked immunosorbent assay, using monoclonal antibody, to detect enterotoxic Escherichia coli K99 antigen in feces of dairy calves

A modified, double-antibody, enzyme-linked immunosorbent assay (ELISA) was developed to detect the K99 pilus antigen of enterotoxic Escherichia coli (ETEC) in feces of calves. Extremely high positive to negative ratios (greater than 200) were obtained by using monoclonal antisera as the primary antibody. Strong positive reactions were obtained with strains of E coli known to produce the K99 antigen; however, non-enteropathogenic E coli (strains not producing the K99 antigen), Salmonella, Proteus, Klebsiella, Pseudomonas, Staphylococcus, Streptococcus, and rotavirus produced negative results. Seventy-five fecal samples, 8 from healthy calves and 67 from calves with neonatal calf diarrhea were examined with the K99 ELISA for the presence of ETEC. Rotavirus test and fecal culture results were available on feces from calves with diarrhea and were used with the K99 ELISA results to determine the specific cause of the disease. Enterotoxic E coli was the predominant agent detected in the feces of 29 diarrheal calves less than 5 days of age. Mixed infections of rotavirus and ETEC were also common in these calves, but rotavirus infections alone were not detected. In 38 calves greater than or equal to 5 days, rotavirus was detected without ETEC. Of these calves, only 2 produced positive tests with the K99 ELISA. Salmonella sp and Proteus sp were detected from 5 of 67 calves with diarrhea.     

The studies on the aetiology of diarrhoea in neonatal calves and determination of virulence gene markers of Escherichia coli strains by multiplex PCR

The purpose of this study was to determine aetiological agents of diarrhoea in neonatal calves and to investigate virulence gene markers of Escherichia coli strains isolated from calves by multiplex polymerase chain reaction (PCR). Eighty-two diarrhoeic calves and 18 healthy calves were used as subjects. Faeces were taken from the rectums of all the calves and were subjected to bacterial culture. Antigen enzyme-linked immunosorbent assay (ELISA) was performed to detect rotavirus, coronavirus and E. coli K99 in faeces of all the calves. A multiplex PCR was used to characterize E. coli strains in all the calves. Escherichia coli was isolated from 37 faeces samples, Enterococcus ssp. was isolated from 22 faeces samples and Salmonella was isolated from one faeces sample in diarrhoeic calves. Furthermore, only E. coli was isolated from all 18 faeces samples of healthy calves. Of the 37 E. coli isolated from diarrhoeic calves, K99 (18.9%), F41 (18.9%), heat-stable enterotoxin a (STa) (18.9%), Shiga toxin 1 (Stx1; 13.5%) and Shiga toxin 2 (Stx2; 5.4%) and intimin (8.1%) genes were identified by multiplex PCR. Of the 18 E. coli isolated from healthy calves, K99 (16.6%) and intimin (55.5%) genes were identified by PCR. A total of 15 rotavirus, 11 coronavirus and 11 E. coli K99 were detected in diarrhoeic calves by the antigen ELISA. As a result, this study shows that rotavirus, coronavirus, E. coli and Enterococcus ssp. were determined to play a role in the aetiology of diarrhoea in the neonatal calves. K99, F41, STa, Stx1 and Stx2 were found as the most common virulence gene markers of E. coli strains isolated from calves with diarrhoea. Multiplex PCR may be useful for characterization of E. coli isolated from calves.     

1株多重耐药致病性大肠杆菌的分离鉴定与分子分群

2017年1月,新疆博乐某牛场犊牛群出现腹泻疫情,本试验旨在调查病因并制定相应的防控措施。采集6头腹泻犊牛的肛拭子和新鲜粪便样本,先用Speed V-Diar4^TM试纸进行轮状病毒、冠状病毒、隐孢子虫和大肠杆菌F5(K99)的检测,再对样本进行病原菌的分离鉴定和分子分群,最后对分离菌进行致病力及耐药性的检测。结果显示,所有粪便样本的试纸检测结果均为阴性,从6份样品中共得到6株大肠杆菌和1株爱德华氏菌,分子分群PCR检测结果显示6株大肠杆菌中A群占83.3%(5/6),F群占16.7%(1/6)。半数致死量结果显示,只有1株大肠杆菌(菌株XJ-B1)具有致病性,可致小白鼠腹泻和死亡。药敏试验结果显示,7株分离菌对这18种抗生素产生不同程度的耐药性,其中致病性大肠杆菌XJ-B1耐药程度最严重,对β-内酰胺类、磺胺类和多肽类抗生素耐药,为多重耐药菌。由此可见,这株致病性大肠杆菌可能是引起本次犊牛腹泻的病因之一,也是本试验首次分离到牛源的F群大肠杆菌,该菌的多重耐药性增加了临床治疗的难度,而本试验结果为该牛场治疗此次犊牛腹泻提供了依据。     

Bovine cryptosporidiosis in Denmark. 2. Cryptosporidia associated with neonatal calf diarrhea

In recent years cryptosporidia have often been identified in diarrheic calves of under one month old, either as the sole pathogenic agent or in combination with other diarrhea-causing infectious agents. In 203 cases of bovine cryptosporidiosis, recorded over a period of two years, cryptosporidia were present in such great numbers that they would seem to be of causal significance. The calves were from three days to five weeks old. In 111 cases (55%) a monoinfection with cryptosporidia occurred, while in the rest of the cases mixed infections with other agents associated with calf diarrhea were demonstrated, especially rotavirus (61 cases) and coronavirus (32 cases); in addition, mixed infections with ETEC (four cases), septicaemic E. coli (two cases), salmonellosis (11 cases), and BVD and coccidiosis (one case each) were seen. In 87 (10%) of 849 4-21-day-old dead calves a massive occurrence of cryptosporidia was demonstrated. Extreme emaciation was noted in 36 of 122 autopsied calves. Most of these calves were more than 14 days old, and mono-infection with cryptosporidia was demonstrated in two thirds of them. In younger calves, more acute cases of cryptosporidiosis were commoner, often in connection with concomitant infections with rota- or coronavirus.     

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).     

Prevalence of four enteropathogens in the faeces of young diarrhoeic dairy calves in Switzerland

The prevalences of Cryptosporidium parvum, rotavirus, bovine coronavirus (bcv), and enterotoxigenic Escherichia coli (E coli k99) were determined in diarrhoeic dairy calves aged one to 21 days on 71 dairy farms in western Switzerland during the winter of 2005 to 2006. Faecal samples from 147 untreated calves suffering from acute diarrhoea were analysed by standardised diagnostic methods, and the immunoglobulin status of each calf was evaluated. The prevalences of C parvum, rotavirus, bcv and E coli k99 were 55.0 per cent, 58.7 per cent, 7.8 per cent and 5.5 per cent, respectively. The proportions of herds positive for the respective pathogens among the herds with diarrhoeic calves were 41.7 per cent, 52.1 per cent, 2.1 per cent and 2.1 per cent. The immunoglobulin concentration in the serum of 90.5 per cent of the diarrhoeic calves was below 8 g/l.     

Reduction in morbidity due to diarrhea in nursing beef calves by use of an inactivated oil-adjuvanted rotavirus-Escherichia coli vaccine in the dam.

An outbreak of neonatal diarrhea occurred among beef calves (2000 animals) from one large Argentinian farm in 1985. Rotavirus was detected in 78% (106/136) and enterotoxigenic Escherichia coli in 1.5% of the samples (2/136) obtained from sick calves. In comparison rotavirus was identified in only 1.6% (1/63) of the samples from clinically healthy calves. The rotavirus strain responsible for the outbreak was characterized as serotype 6 belonging to group A. In the following three years the protective capacity of a combined rotavirus-E. coli inactivated vaccine administered to the dams during the last third of the gestation period was evaluated on this farm by comparison of morbidity due to diarrhea in calves from vaccinated vs. placebo cows within the same year. The morbidity due to diarrhea among calves from dams in the vaccinated and placebo groups was 34% and 77%, respectively in 1986; 23% and 47% in 1987, and 15% and 34%, in 1988. In 1987 morbidity of diarrhea in calves born from vaccinated heifers was 54% and 74% in calves from placebo heifers. In 1988 morbidity from diarrhea was 41% and 54%, respectively among calves in these two groups. In all experiments, calves from heifers showed significantly greater morbidity than calves from cows. Differences in diarrhea morbidity between the vaccinated and placebo groups were statistically significant (P less than 0.05). Additional studies showed that the diarrhea had a significant influence (P less than 0.05) on the average live weight of the calves at weaning (5 to 7 months) with an average weight loss of 7.8 kg per calf among the calves affected with diarrhea.     

Effect of combined rotavirus and Escherichia coli in neonatal gnotobiotic calves

Gnotobiotic calves (24 hours old) were monoinfected with calf rotavirus (CRV) strain NCDV, an enterotoxigenic Escherichia coli (ETEC) strain B44 (K99+), or a nonenterotoxigenic E coli (NETEC) strain 123 (K99-). Calves also were dually infected with CRV and either ETEC or NETEC. Eighteen calves equally allotted between 6 treatment groups were used in these studies: Noninfected controls--group A; CRV--group B; ETEC--group C; NETEC--group D; CRV + ETEC--group E; and CRV + NETEC--group F. Severe diarrhea and villous atrophy were observed in calves of treatment groups B, C, E, and F. Mortality was present only in treatment groups C and E as result of ETEC infection. There were no significant differences in the clinical responses or enteric lesions between treatments B and F, although a significant increase in the concentrations of NETEC was demonstrated in calves dually infected with CRV + NETEC (group F) as compared with calves monoinfected with NETEC (group D). Calves inoculated with ETEC (group C) had severe villous atrophy, neutrophilic infiltration of intestinal lumen, and moderate enterocyte necrosis. Calves dually inoculated with CRV + ETEC (group E) had the most extensive and severe lesions, similar to those in group C, plus a pronounced necrotic fibrino-hemorrhagic enteritis. Infection of enterocytes by CRV did not affect in any way the adherence of ETEC to the intestinal mucosa. Dual viral and bacterial infections of the same enterocytes were evident.     

An epidemiological study of selected calf pathogens on Holstein dairy farms in southwestern Ontario.

Fecal samples from calves on 78 randomly selected Holstein dairy farms in southwestern Ontario were screened for Salmonella, Campylobacter jejuni/coli, enteropathogenic Escherichia coli, rotavirus and coronavirus. Based on the observed prevalence, 22% of farms had calves infected with Salmonella, 13% with Campylobacter jejuni/coli, 41% with enteropathogenic E. coli, 19% with rotavirus and 5% with coronavirus. These estimates can be modified, using a method developed by Mullen and Prost (1983) for the World Health Organization, to account for the nature of the laboratory test used. If the test is assumed to have no false positives (that is, if an organism is detected it must be there), then the observed prevalence estimates seen on this study may greatly underestimate the true prevalence of infected premises. The use of nipple feeders for calves was associated with an increased probability of farms having calves shedding detectable fecal levels of Salmonella, E. coli, or one of the two viruses. The use of group pens was associated with an increased odds of finding C. jejuni. Calves with diarrhea on these farms tended to have increased odds of shedding rotavirus, and E. coli with the K99 antigen. However, at the farm level, none of the organisms was associated with above median levels of morbidity. Farms positive for one or other of the viruses had increased odds of experiencing calf mortality relative to virus-negative farms, and farms positive for C. jejuni/coli had decreased odds of mortality.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of microbial and host variables on the interaction of rotavirus and Escherichia coli infections in gnotobiotic calves

Naturally occurring mixed infections with Escherichia coli and rotavirus have been associated with fatal diarrhea of calves about 1 week old. Experiments were designed to reproduce this syndrome in gnotobiotic calves. Clinical, microbiological, and pathologic data were used to assess severity of disease and mechanisms of the interaction between the 2 infections. An initial study involved 5- to 8-day-old gnotobiotic calves inoculated with a strain of enterotoxigenic E coli (ETEC) and a strain of rotavirus. Calves were observed for 2 days after they were inoculated; fatal diarrhea was not produced. In later studies, variables were tested to identify those that might contribute to fatal diarrhea. Variables which did not result in fatal or severe diarrhea or which did not cause disease that was more severe in dually inoculated calves than that in monoinoculated calves were increasing feed to 2 times base line, increasing dose of ETEC to 10 times base line, inoculating calves when they were 2 days old, using a strain of E coli that causes colisepticemia, and using a different strain of rotavirus. When the observation period was extended from 2 days to 6 days after calves were inoculated, severe, watery, fatal diarrhea occurred in 6 of 12 calves by 32 to 72 hours after dual inoculation was given. Fatal diarrhea was associated with intensive colonization by the ETEC in the caudal half of the small intestine. Microscopic lesions were similar between dually inoculated and rotavirus-monoinoculated calves, except there was more severe atrophy of ileal villi of dually inoculated calves.(ABSTRACT TRUNCATED AT 250 WORDS)     

Occurrence and properties of FY(Att25)+ Escherichia coli associated with diarrhoea in calves

Escherichia coli producing the adhesive antigen FY(Att25) were isolated from 46 of 1341 (3.4 per cent) E coli isolated from calves on 20 of 164 (12.1 per cent) farms in Scotland and England. Twenty of the 46 calves had diarrhoea and in nine of these animals there were mixed infections with rotavirus, coronavirus, cryptosporidium and Salmonella typhimurium. The F41 fimbrial adhesin was found on one of the FY(Att25)+ E coli. This strain also produced heat stable enterotoxin. The remaining FY(Att25)+ isolates did not produce other adhesins, enterotoxins or verotoxins. The FY(Att25) antigen was not detected on 109 pathogenic E coli isolated from calves, chickens, lambs, pigs or man.     

Development of monoclonal antibody ELISA for simultaneous detection of bovine coronavirus, rotavirus serogroup A, and Escherichia coli K99 antigen in feces of calves.

A rapid ELISA was developed for simultaneous detection of bovine coronavirus (BCV), rotavirus (RV) serogroup A, and Escherichia coli K99 antigen in feces of calves. A mixture of 3 monoclonal antibodies specific for BCV, RV, or K99 was used successfully to capture the antigens; the same antibodies labeled with peroxidase were used to detect BCV, RV, or K99. The triple ELISA was compared with standard reference diagnostic methods by examining feces from experimentally and naturally infected and healthy calves. All the components of the test were highly specific (greater than 90%) and sensitive (BCV, 77%; K99, 93%; RV, 100%) when used in a format requiring short incubation steps at 20 C and visual recording of results.     

Cryptosporidiosis in neonatal calves: 277 cases (1986-1987)

From January 1986 through December 1987, 277 cases of cryptosporidiosis in calves were diagnosed by the South Dakota State University Diagnostic Laboratory. Cryptosporidium sp was the only pathogen identified in 142 (51.3%) of the calves. Concurrent infections with rotavirus, coronavirus, Escherichia coli, Salmonella sp, bovine viral diarrhea virus, or other pathogens were identified in the remaining 135 calves. After elimination of cases involving autolyzed specimens or calves with chronic diarrhea, records of 11 calves with acute, severe cryptosporidiosis were identified in which Cryptosporidium sp was the only pathogen.     

牛病毒性腹泻病毒、大肠杆菌和奇异变形杆菌混合感染致犊牛腹泻的研究

为确定甘肃省临夏州某奶牛场犊牛腹泻的病因,并提供合适的治疗方案和防控措施,试验采集该牛场13头腹泻犊牛的粪便和血清,通过胶体金技术、ELISA方法、细菌分离鉴定、Kirby-Bauer法分别进行病毒病原学检测、病毒血清学抗体检测、病原菌鉴定和药物敏感性试验。病毒学检测结果显示,13份粪样中未检测出牛轮状病毒（BRV）、牛冠状病毒（BCV）的抗原,牛病毒性腹泻病毒（BVDV）抗原阳性率为23.08%（3/13）;未检出BRV和BCV的抗体,BVDV血清学抗体阳性率为38.46%（5/13）。病原菌检测结果显示,13份粪便样品中,分离出13株大肠杆菌和7株奇异变形杆菌。药敏试验表明,分离的大肠杆菌和奇异变形杆菌对20种常规药物均产生了不同程度的耐药,且无对两种细菌均有效的药物。此次犊牛腹泻是由BVDV、大肠杆菌、奇异变形杆菌混合感染引起的,且大肠杆菌和奇异变形杆菌的耐药现象严重,本试验结果为该牛场进一步治疗此次的犊牛腹泻病提供了合理有效的依据。     

Diarrhoea in dairy calves reduced by feeding colostrum from cows vaccinated with rotavirus

Forty-two dairy calves remained with their dams for two days after birth, and then were removed to a calf rearing shed. Calves were allocated to three groups for the next 14 days, and received twice daily either whole milk, whole milk with a 10 per cent supplement of pooled normal bovine colostrum or whole milk with 10 per cent supplement of colostrum from cows vaccinated with rotavirus. A natural outbreak of diarrhoea occurred, affecting 28 of the 42 calves. Feeding immune colostrum delayed the onset of diarrhoea, and reduced its incidence, duration and severity. Live weight gains were consequently improved. The group fed normal colostrum had diarrhoea intermediate in severity between that of control calves and those fed immune colostrum. The aetiology of the diarrhoea was complex, with calves excreting rotavirus, enteropathogenic Escherichia coli and cryptosporidia.     

Prevalence of enteric pathogens in the feces of healthy beef calves

Fecal specimens from 136 healthy beef calves (1 day to 12 weeks of age) were examined for the presence of infectious agents known to cause enteric disease in calves. The calves were selected from 22 herds in which all calves were free of clinically apparent enteric disease. Salmonella sp, enterotoxigenic Escherichia coli, Cryptosporidium, and coronavirus were not detected in any of the calves. Three calves were infected with rotavirus and 1 calf was infected with Yersinia enterocolitica. Campylobacter-like bacteria were isolated from 50 of 130 calves, with 36 of the calves positive for C jejuni. Seemingly, clinically normal calves may be infected more often with enterotoxigenic E coli, Cryptosporidium, coronavirus, or rotavirus in herds in which some calves have enteric disease than in herds free of major enteric disease. Campylobacter jejuni was well adapted to the bovine host and was of similar prevalence in diarrheal and nondiarrheal calves. The K99 positive, nonenterotoxigenic E coli was isolated from the feces of 16 healthy calves. Information in addition to the presence of K99 antigen is useful when diagnosing enterotoxic colibacillosis in calves.     

Neonatal calf diarrhea caused by a virus that induces villous epithelial cell syncytia.

Intestinal lesions caused by a virus serologically unrelated to the calf diarrheal rotavirus or coronavirus were studied in gnotobiotic calves. The virion purified from feces from infected calves was a fringed particle with a diameter of about 100 nm. The incubation period from time of inoculation per orum to onset of diarrhea in calves was as short as 8 hours. The viral infection in bacteria-free calves or calves not contaminated with pathogenic bacteria caused severe illness for only 24 hours. When bacteria such as the K99 antigen Escherichia coli were present, the combined infection caused mortality. Lesions occurred only in the small intestinal villous epithelium. Calves euthanatized shortly before or after the onset of diarrhea had developed villous epithelial cell syncytia that contained numberous virions in the cytoplasm. Within 2 to 3 hours after onset of diarrhea, the infected cells were shed and the villi had denuded tips or had cuboidal to squamous epithelial cells.     

一例冠状病毒、轮状病毒混合感染引起犊牛腹泻的诊治

2022年3月,河南省某规模化奶牛养殖场发生一起新生犊牛腹泻病例,在流行病学调查、临床症状观察、病理剖检的基础上,采集6头腹泻犊牛的新鲜粪便进行了寄生虫卵检查、小球隐孢子虫、牛轮状病毒、冠状病毒、大肠杆菌K99抗体检测,血液样品进行了牛病毒性腹泻病毒抗原检测,组织样品进行了细菌分离。结果表明牛轮状病毒抗原6头阳性,阳性率100%;冠状病毒抗原4头阳性、2头阴性,阳性率66.7%;其他病原均为阴性。根据临床症状、病理变化和实验室检测结果,确诊该病例为牛冠状病毒和轮状病毒混合感染引起的犊牛腹泻。根据诊断结果,采取了改善饲养管理、补液、收敛、止泻等综合性防治措施,疫情得到了有效的控制,为临床防治犊牛腹泻提供了参考。