Microbiology of calf diarrhoea in southern Britain

Faeces samples from calves with diarrhoea in 45 outbreaks were examined for six enteropathogens. Rotavirus and coronavirus were detected by ELISA in 208 (42 per cent) and 69 (14 per cent) of 490 calves respectively; calici-like viruses were detected by electron microscopy in 14 of 132 calves (11 per cent). Cryptosporidium were detected in 106 of 465 (23 per cent), Salmonella species in 58 of 490 (12 per cent) and enterotoxigenic Escherichia coli bearing the K99 adhesin (K99+ E coli) in nine of 310 calves (3 per cent). In the faeces of 20 per cent of calves with diarrhoea more than one enteropathogen was detected; in 31 per cent no enteropathogen was found. Faces samples from 385 healthy calves in the same outbreaks were also examined. There was a significant statistical association of disease with the presence of rotavirus, coronavirus, Cryptosporidium and Salmonella species (P less than 0.001). Healthy calves were not examined for calici-like viruses and the association of K99+ E coli with disease was not analysed because there were too few positive samples. Rotavirus infections were more common in dairy herds and single suckler beef herds whereas Salmonella infections were more often found in calf rearing units. Cryptosporidium were more common in single and multiple suckler beef herds. K99+ E coli were found in one dairy herd and one multiple suckler beef herd both with unhygienic calving accommodation. Variations in coronavirus detection among different farm types were not statistically significant. In this survey rotavirus was the most commonly detected agent in calf diarrhoea and Cryptosporidium were found in approximately one quarter of affected calves. Infection with Salmonella species was widespread, but K99+ E coli infections were less common in the United Kingdom than in other countries.     

Persistent excretion of rotavirus by pregnant cows

In a study of the epizootiology and prevalence of enteropathogens which may be involved in neonatal calf diarrhoea, 10 in-calf cows from a herd with a history of rotavirus-induced calf diarrhoea were monitored over a period of six to seven months. All the cows excreted rotavirus intermittently without showing any clinical signs, and 21.8 per cent of faecal samples contained rotavirus. Reoviruses were isolated from 87 per cent of the samples from the cows, and from all the 10 calves born to them. However, rotavirus was detected in only one calf, and diarrhoea developed only in this calf even though the calves were housed in communal pens. Campylobacter jejuni was isolated from six of the 10 dams and from five of the 10 calves, not including the calf with diarrhoea. Other potential enteropathogens such as cryptosporidium, salmonella, Clostridium difficile, coronavirus and other viruses were not found, but two cows and two calves shed enterotoxigenic Escherichia coli.     

Rotavirus and concurrent infections with other enteropathogens in neonatal diarrheic dairy calves in Spain

Faeces samples from 218, one to 30 days old, diarrheic dairy calves in 65 dairy herds were screened for the presence of rotavirus and concurrent infections with coronavirus, Cryptosporidium, F5+ Escherichia coli and Salmonella spp. Calves were grouped according to their age as follows: 1-7, 8-14, 15-21 and 22-30 days. Rotavirus infection was detected in 46.9%, 45.6%, 33.8% and 48.3% of the calves in the respective age-groups. No significant differences in the detection rate of rotavirus were found among calves on the different age-groups. Rotavirus was the only enteropathogen detected in 39 of the 93 (41.9%) diarrheic calves positive to this agent. Concurrent infections with other enteropathogen(s) were detected in 31.3%, 33.3%, 20.6% and 3.4% of the rotavirus infected calves in the age-groups 1-7, 8-14, 15-21 and 22-30 d, respectively. A significant age-associated decrease in the detection rate of mixed infections (p < 0.01) was found. The detection rates of the other enteropathogens considered in calves with rotavirus infection were 20.4% for coronavirus, 85.2% for Cryptosporidium, 16.7% for F5+ E. coli and 1.8% for Salmonella.     

Aetiology of diarrhoea in young calves

Faeces samples were collected from 302 untreated calves on the day of onset of diarrhoea and from 49 healthy calves at 32 farms experiencing outbreaks of diarrhoea. At least four diarrhoeic calves were sampled on each farm, and samples were examined for rotavirus, coronavirus, cryptosporidium, enterotoxigenic Escherichia coli and Salmonella species. Although all these enteropathogens were excreted more frequently by the diarrhoeic than by the healthy calves, the difference was significant overall only for rotavirus. Rotavirus was excreted by 18 per cent of healthy calves, coronavirus by 4 per cent, cryptosporidium by 14 per cent, and no enterotoxigenic E coli or Salmonella species were detected. The most common enteropathogen in diarrhoeic calves was rotavirus, which was excreted by more than half the diarrhoeic calves on 18 farms. Coronavirus was excreted at a similar high prevalence on one farm, cryptosporidium on five farms and enterotoxigenic E coli on three farms. Concurrent infection with two or more microorganisms occurred in 15 per cent of diarrhoeic calves. There was no difference in the isolation rate of campylobacters between diarrhoeic and healthy calves.     

Agents associated with neonatal diarrhoea in Ethiopian dairy calves

Faeces samples collected from diarrhoeic dairy calves in the first 8 weeks of life were examined for the presence of 5 enteropathogens. The majority of the 108 diarrhoea cases occurred in the first 5 weeks of life and a commercial ELISA kit detected bovine enteric coronavirus (BEC) in 38.9%, serogroup A rotavirus (RV) in 16.7% and K99 (F5) fimbrial adhesin-positive Escherichia coli (K99 ETEC) in 11.1 per cent. Concurrent infections of these enteropathogens were detected in 14.8% of samples (30.8% of samples positive for these agents). No evidence of cryptosporidial infection was found using a differential staining method on faecal smears nor was salmonella excretion detected. On 2 of the 8 farms only BEC was present; the other 6 farms were positive for all 3 agents. It is concluded that BEC is the major infectious cause of neonatal calf diarrhoea in the Ethiopian dairy herds studied with RV and K99 ETEC also contributing to morbidity, either alone or as mixed infections.     

Isolation of coronaviruses from neonatal calf diarrhoea in Great Britain and Denmark

Coronaviruses were isolated from neonatal calves with diarrhoea in Great Britain and Denmark. They were serially passed in gnotobiotic calves which developed acute diarrhoea. Pathological lesions were found in the small and large intestines. Coronaviruses were demonstrated by electron microscopic examination of the faeces and intestinal contents, immunofluorescent staining of sections of small and large intestine and by isolation in tracheal organ cultures. In early passages of the British coronavirus, particles of about 30 nm in diameter were observed in the faeces by electron microscopy. These particles were removed from the coronavirus preparations by cross-protection experiments in calves. The coronaviruses were morphologically and antigenically similar to the bovine coronavirus isolated in the United States and the British virus was adapted to replicate in calf kidney cell cultures.     

Rotavirus infections in calves in dairy herds

Neonatal calf diarrhoea was studied in 115 calves of one dairy herd from January 1976 to June 1977. Two syndromes could be distinguished: a mild and short lasting 'early diarrhoea' within the first three days of life and a usually more severe 'late diarrhoea' from the fourth to the 14th day of life. The latter type of diarrhoea occurred almost exclusively during the first half of the year. Thirty-four out of 45 calves with late diarrhoea excreted rotaviruses, whereas only one of 34 calves with early diarrhoea excreted virus. In addition, rotavirus excretion was found in 11 calves that either remained healthy or had recovered from diarrhoea before virus was first detected. Similar findings were obtained in the spring of 1978 but in addition bovine coronavirus was recovered from four calves with late diarrhoea and from three healthy calves. One faecal sample obtained from a calf with diarrhoea on day 2 yielded K99+ Escherichia coli. Nearly all cows excreted rotavirus-specific antibodies in their colostrum but no relationship was found between the initial colostral antibody titre against rotavirus and the development of rotavirus-associated diarrhoea in the calf.     

Rotavirus and coronavirus associated diarrohoea in calves

A virological survey was carried out on 150 calf faeces from 30 central North Island properties, utilising electron miscroscopic examination of ultracentrifuged faecal pellets. Rotaviruses were detected on three farms, and coronavirus-like particles on two farms.

In the same area, three other properties with past histories of rotavirus infection were also studied. Weekly examination of faeces from 10 calves from each property over a period of 5 weeks demonstrated that rotavirus excretion commenced at the same time as the onset of outbreaks of diarrhoea. On two of these rotavirus-positive farms, coronaviruses were also detected. At one farm, this coincided with rotavirus excretion and diarrhoea, and was associated with a more severe clinical disease but, on the other property, the coronavirus was unassociated with clinical disease. A similar study, carried out on a further property with a past history of coronavirus infection, showed that coronavirus excretion commenced simultaneously with the onset of an outbreak of diarrhoea. The virus was, however, detected in small numbers in 1 of 10 calves only.

The results of the survey, coupled with previously gathered data, indicate that both viruses may be widespread in the cattle population. On some properties, rotavirus had a clear association with diarrhoea, but the relationship of coronavirus to diarrhoea was less well-established.     

Studies on neonatal calf diarrhoea caused by rotavirus: transmission of the disease and attempted vaccination of colostrum-deprived calves

Mild to severe scouring could be produced in colostrum-deprived calves with tissue culture-adapted rotavirus and feacal material from field cases of calf diarrhoea. The feaces of experimentally infected calves contained rotavirus for at least 3 days. Pathogenic bacteria were presented in one calf only and this calf also showed the most severe gastroenteritis. Eight calves were vaccinated with a live rotaviral calf diarrhoea vaccine and subsequently challenged with infective rotavirus. Mild scouring was observed after vaccination, but the calves remained normal after challenge. Rotavirus particles were detectable in the faeces for a few days after vaccination and challenge.     

Evaluation of a combined rotavirus and enterotoxigenic Escherichia coli vaccine in cattle

A vaccine of rotavirus and K99 antigen from enterotoxigenic Escherichia coli was emulsified in oil adjuvant and administered intramuscularly to pregnant cows. Calves born to and reared on vaccinated dams were protected against experimental rotavirus infection at five days old when compared with calves from unvaccinated control cows. Field trials of the vaccine were carried out in 40 commercial herds, in which half the cows in each herd were selected at random for vaccination and half were left unvaccinated. In 31 herds (2641 cows) there was no significant diarrhoea problem (less than 10 per cent morbidity); these herds were excluded from further analysis. The nine remaining herds did experience a calf diarrhoea problem of greater than 10 per cent morbidity, but on four farms the disease was associated with cryptosporidiosis and on a fifth no enteropathogens were detected; these five farms (461 cows) were also excluded from further analysis. Of the remaining four herds, two beef suckler herds (105 cows) had concurrent rotavirus and cryptosporidial infections, and vaccination was associated with a decreased excretion of rotavirus but not with a decreased incidence of diarrhoea. In the other two dairy herds (68 cows) with prevaccination rotavirus problems, there was a significantly decreased incidence of diarrhoea in calves born to vaccinated cows. No natural field challenge of enterotoxigenic E coli was encountered on any of the trial farms.     

Prevalence,prediction and risk factors of enteropathogens in normal and non-normal faeces of young Dutch dairy calves

Between January and April 2007, 424 calves under 22 days of age from 108 Dutch dairy herds were sampled to estimate the prevalence of non-normal faeces (‘custard-like’—yellowish-coloured with custard consistency or diarrhoea: watery-like faeces) and the shedding of enteropathogens Escherichia coli K99 (E. coli), Coronavirus, Cryptosporidium parvum (C. parvum), Rotavirus and Clostridium perfringens (Cl. perfringens). In addition, information was collected on animal characteristics and herd-management practices. The probability of detecting each one of five enteropathogens given a calf with ‘custard-like’ faeces or diarrhoea was estimated using Bayes’ rule and was based on the predicted probabilities from a multinominal model including each of five enteropathogens as independent variables. In addition, putative risk factors for the presence of each of five enteropathogens were analysed using logistic regression models with random herd effects.Fifty-seven percent of calves had faeces of normal colour (brownish) and consistency (firm), 23.8% (95%CI: 19.8–28.2%) had ‘custard-like’ faeces and 19.1% (95%CI: 15.5–23.2%) had diarrhoea. E. coli was the least detected enteropathogen (2.6% (95%CI: 1.3–4.6%) of calves, 9% (95%CI: 5–16%) of herds) and Cl. perfringens was most detected (54.0% (95%CI: 49.1–58.8%) of calves, 85% (95%CI: 77–91%) of herds). E. coli and Coronavirus were detected incidentally in only one or two calves per herd, whereas C. parvum and Cl. perfringens were frequently detected in up to four calves per herd. For calves with ‘custard-like’ faeces, the probability of detecting Rotavirus from a calf in its first week of age was 0.31 whereas for a calf in its second week, there was a 0.66 probability of detecting C. parvum. The probabilities of detecting E. coli, Rotavirus and C. parvum in calves with diarrhoea in their first week of age were 0.10, 0.20 and 0.43, respectively. In calves with diarrhoea between 1 and 2 weeks of age, the probability of detecting enteropathogens was 0.43 for C. parvum. None of the tested enteropathogens were related to ‘custard-like’ faeces or diarrhoea in the third week of age.Putative risk factors for E. coli, Coronavirus and C. parvum included the presence of peer-calves shedding Coronavirus, C. parvum or Rotavirus, respectively. Additionally, managerial risk factors such as non-optimal hygienic housing (for Coronavirus) and the routine use of antibiotics for diarrhoeic calves (for C. parvum) were found. No animal or managerial factors were associated with shedding of Cl. perfringens.     

Advances in prevention and therapy of neonatal dairy calf diarrhoea: a systematical review with emphasis on colostrum management and fluid therapy

Neonatal calf diarrhoea remains the most common cause of morbidity and mortality in preweaned dairy calves worldwide. This complex disease can be triggered by both infectious and non-infectious causes. The four most important enteropathogens leading to neonatal dairy calf diarrhoea are Escherichia coli, rota- and coronavirus, and Cryptosporidium parvum. Besides treating diarrhoeic neonatal dairy calves, the veterinarian is the most obvious person to advise the dairy farmer on prevention and treatment of this disease. This review deals with prevention and treatment of neonatal dairy calf diarrhoea focusing on the importance of a good colostrum management and a correct fluid therapy.     

Longitudinal survey of rotavirus infection in calves

A longitudinal survey of rotavirus infection in heifer calves was carried out on a closed Friesian dairy herd over two successive calving seasons. Rotavirus was detected by electron microscopy in the faeces of 45 of 57 (79 per cent) calves examined. On average the virus was first detected at 6.1 days of age. Clinically the disease associated with rotavirus infection was of mild to moderate severity. Only one infected calf required intravenous fluid therapy. Diarrhoea or excretion of abnormal faeces was associated with rotavirus infection in 58 per cent of infected calves, while in the remaining 42 per cent infection was subclinical. The cycle of rotavirus infection was broken by thorough cleansing and disinfection of the calf house.     

Dysentery in calves caused by an atypical strain of Escherichia coli (S102-9)

Dysentery lasting 4-8 days was produced in five 4-day-old colostrum-fed calves, after inoculation with an atypical strain of Escherichia coli S102-9; peak excretion of S102-9 occurred during the period of dysentery. Two calves were killed when clinical signs were most severe and bacteria were seen attached to the surfaces of enterocytes in the large intestine; microscopic lesions were seen in these areas. The lesions were identical to those previously reported in a natural outbreak of dysentery in calves, from which E. coli S102-9 was isolated, and to those seen in gnotobiotic calves experimentally infected with S102-9. Reinfection of the three surviving calves 16-20 days later with S102-9 and primary infection of two calves aged 24 and 51 days did not cause dysentery. Four of 659 coliforms isolated from field outbreaks of calf diarrhoea resembled the atypical strain S102-9. These four isolates and S102-9 did not produce heat-stable enterotoxin, but all produced a toxin cytopathic for Vero and HeLa cells. Two of the four isolates were inoculated alone into 4-day-old gnotobiotic calves deprived of colostrum; neither calf developed dysentery but microscopic lesions identical to those produced by S102-9 were detected in the large intestines of both animals.     

Electron microscopic detection rate of enteral viruses in diarrhea of dogs, cats, calves, swine and foals in the year 1988--electron microscopic study results

During 1988 fecal and gut samples of 641 dogs, 198 cats, 576 calves, 108 piglets and 64 foals with diarrhoea were investigated for virus infections by electron microscopy. In samples of dogs and cats parvovirus was detected at a proportion of 21.9% and 16.7%, respectively; rotavirus alone or together with coronavirus was found only in 0.3-1.5% of the specimens. In samples of calves rotavirus, as well as coronavirus dominated with a detection rate amounting to 17.4% and 26.6% respectively (including 4.5% of mixed infections); parvovirus was present in a ratio of 0.5%. Specimens of piglets mainly contained coronavirus (25.0%), and in lower percentages rotavirus (2.8%), rota- and coronaviruses (0.9%) and parvovirus (0.9%). In feces of foals rotavirus was detected in 6.3% and particles resembling picornavirus in 4.7% of cases. Not identifiable virus particles resembling corona-or picornaviruses were rarely found (between 0.6-2.5) also in specimens of the other animal species.     

Studies with an unclassified virus isolated from diarrheic calves

A transmissible agent (Breda agent) was isolated from a calf with diarrhea and shown to be infectious by inoculation orally into gnotobiotic and conventionally reared calves. The “Breda” agent had the morphology of a virus and possessed a hemagglutinin. Antigenic studies showed the virus to be antigenically different from bovine coronavirus, parainfluenza 3 virus, bovine rotavirus, bovine parvovirus and bovine pestivirus (BVD). Attempts to culture the virus in cell or organ cultures or in embryonated eggs, were unsuccessful. The virus was either spherical or kidney shaped, with 7–9 nm peplomers on the surface. A few particles possessed coronavirus processes of 17–20 nm, but these were arranged irregularly and were thought to be tissue debris. Three out of eight experimental calves developed severe diarrhea and the lesions in the small and large intestines were similar to those reported for coronavirus. The virus replicated in the jejunal and ileal regions of the small intestine and in the spiral colon, as judged by immunofluorescence. The virus multiplied in all experimental calves and was excreted in the feces; excretion correlating with the onset of diarrhea or a change in the appearance of the feces. There was little or no malabsorption measured by the uptake of D-xylose and the fact that infection of both the crypt and villus epithelial cells was observed, suggests that the pathogenesis may be different from rotavirus and coronavirus. Fourteen of fortyseven calves in the outbreak were infected with the virus, virus was not identified in other farm outbreaks of the disease.     

Cryptosporidium parvum and Giardia intestinalis in calf diarrhoea in Sweden

The objective of this study conducted in 75 herds was to investigate the presence and significance of Criptosporidium parvum and Giardia intestinalis in Swedish dairy calves in comparison with rotavirus, coronavirus and Escherichia coli K99+. The farmers were asked to collect faecal samples from each heifer calf that had diarrhoea between birth and 90 days of age, and also from a healthy calf of the same age. In total, 270 samples were collected and analysed. C. parvum, either alone or together with G. intestinalis and/or rotavirus, was detected in 16 (11%) and 6 (5%) of the samples from diarrhoeic and healthy calves, respectively. Even though a higher proportion of diarrhoeic calves shed C. parvum, the difference between the groups was not statistically significant (p = 0.067), possibly due to the low number of positive samples. G. intestinalis was found in 42 (29%) of the diarrhoea samples and in 29 (23%) of the samples from healthy calves. Rotavirus and coronavirus were demonstrated in 24% and 3% of the diarrhoea samples, respectively, whereas E. coli K99+ was only found in samples from 2 healthy calves. C. parvum and G. intestinalis were found in samples from calves 7 to 84 days of age and during all seasons. The results confirm that C. parvum is present in Swedish dairy herds and might have clinical significance. G. intestinalis was the most common agent found but the importance of this parasite remains unclear. Both parasites have suggested zoonotic potential and thus warrant further attention. In addition, rotavirus is a major pathogen in neonatal enteritis in Sweden, whereas coronavirus and E. coli K99+ seem to be of less importance.     

Enteric infections in veal calves: a longitudinal study on four veal calf units

Forty-five calves on four veal calf units were monitored during the first four weeks after their arrival. Faecal samples were collected on alternate days and screened for the presence of rotaviruses, bovine coronavirus, Cryptosporidium oocysts, K99 positive strains of E. coli and Salmonella spp. Rotaviruses and Cryptosporidium were the most commonly detected agents (78% and 60% respectively of the calves). Bovine coronavirus was detected in the faeces of 18% of the calves, whilst K99 positive E. coli was only found in 2 samples from one calf. Salmonella spp. were not isolated from any of the 646 faecal samples examined. Shedding of rotaviruses occurred in a bimodal pattern beginning in the first week of the survey. Cryptosporidium oocysts were detected most frequently in the interval between the two peaks of rotavirus shedding. The presence of rotaviruses or Cryptosporidium oocysts in faeces was not strongly associated with scour, nor were combined infections with these agents or the cases of bovine coronavirus infection. The condition of the calves throughout the survey was generally satisfactory.     

Rotavirus and enterotoxigenic Escherichia coli infections of calves on a closed Finnish dairy farm

Rotavirus and enterotoxigenic Escherichia coli infections of calves were surveyed during 2 successive years on a closed Finnish dairy farm consisting of 90–105 milking cows. From a clinical standpoint, diarrhoea was of moderate to high severity during the first year, compared to the milder disease in the second year of the study. Diarrhoea or abnormal faeces were found only in calves less than 8 weeks old, with the peak occurring during the first 2 weeks of life.In the first year, rotavirus was detected throughout the calving season in diarrhoeic or abnormal faeces of calves aged 1 day to 7 weeks. In the second calving season, rotavirus was detected only during the 4 autumn months and in calves aged 11 days to 8 weeks. Rotavirus was detected in only 1 sample of normal faeces in both years. Electron microscopy revealed no enteropathogenic viruses other than rotaviruses. Enterotoxigenic K99 E. coli was found in about half of diarrhoeic or abnormal faeces in both years and throughout the calving seasons. K99 E. coli was also found in 5–10 % of normal faeces.In the second year of the study, 45 of 101 pregnant dams were vaccinated with 2 doses of E. coli antigen. The vaccination trial did not prevent or reduce altered faeces in calves whose dams had been vaccinated compared with calves whose dams had not been vaccinated in the same year. Comparing the 2 years, the earlier uptake of colostrum together with better cleaning and disinfection of the calf house, contributed to the later and rarer occurrence of rotavirus infection in the second year of the study. The earlier uptake of colostrum together with better cleaning and disinfection of the salf house, in the second year, could not prevent enterotoxigenic E. coli infections in calves but partly prevented and modified the disease.     

Diagnosis of bovine coronaviruses using mouse erythrocyte suspensions stabilized with formalin

Bovine coronavirus isolated from calf faeces diseased with gastroenteritis and passaged to colostrum-free calves agglutinated mouse and rat erythrocytes. The agglutination reaction depended on temperature and took place only at a temperature of 4 degrees C. At a temperature of 37 degrees C the agglutinate broke down within 15 minutes. The coronavirus could be detected by the haemagglutination test in the contents of the small and large intestines and in the faeces of experimentally and naturally infected calves. The agglutination capacity of mouse erythrocytes was not affected by careful fixation of these erythrocytes with formalin and subsequent lyophilization and remained unchanged for as long as 52 weeks of storage at a temperature of 4 degrees C. It was demonstrated by a comparative examination of 182 samples of the faeces of calves suffering from diarrhoea that haemagglutination test was as sensitive as electron microscopy.