Virus and virus-like particles in the faeces of cats with and without diarrhoea

Negative staining electron microscopy was used to identify viruses in 166 normal and 62 diarrhoeal faecal samples from 208 cats admitted to an animal shelter during a 16-month period (March 1984 to June 1985). On the basis of size and shape 7 distinct viral types were detected: 24 nm parvovirus-like particles, 30 nm astrovirus, 30 nm picornavirus-like particles, reovirus, rotavirus, coronavirus and a 75 nm "togavirus-like" particle. The incidence of these particles in the 208 cats was 11%, 7%, 6%, 0.4%, 5%, 1% and 1% respectively. Virus isolation studies using 40 of the faecal samples succeeded in isolating reovirus 1 in 2 cases. Immune electron microscope studies demonstrated the presence of antibody in a human serum to cat astrovirus, but failed to clarify the identity of the parvovirus-like particles and picornavirus-like particles, other than showing that some of the parvovirus-like particles were not related to feline panleukopenia virus. It was found that parvovirus-like particles, astrovirus, picornavirus-like particles, reovirus and rotavirus could be excreted by cats with normal faeces as well as cats with diarrhoeal faeces. Parvovirus-like particles, astrovirus, picornavirus-like particles and rotavirus could be excreted in high concentration in normal faeces. There was no simple relationship between age and diarrhoea in the population of cats studied. Age was not a critical factor in the excretion of parvovirus-like particles, astrovirus, picornavirus-like particles and rotavirus. The incidence of diarrhoea was not clearly associated with the seasons.     

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.     

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.     

Winter dysentery in dairy herds: electron microscopic and serological evidence for an association with coronavirus infection.

Faeces and, or, paired sera were collected from cows in six dairy herds with classical winter dysentery. Similar samples were collected from cows in three other dairy herds experiencing non-haemorrhagic diarrhoea during the survey period. Coronavirus was the only enteric pathogen identified by immune electron microscopy (IEM) in all six outbreaks, occurring in 26 of 29 (90 per cent) of the affected cows and in one of 11 normal cows from the same herds. Nineteen of 26 affected cows (73 per cent) developed greater than four-fold increases in neutralising antibody titres to the Mebus strain of bovine coronavirus, compared with two of eight normal cows in the same herds. No cows showed greater than four-fold increases in antibody titres to bovine virus diarrhoea virus. None of the cows from the three herds with non-haemorrhagic diarrhoea shed coronavirus in faeces detectable by IEM or developed greater than two-fold rises in coronavirus antibody titres in paired sera. No enteric pathogens were identified in two of the herds. However, two cows in the third herd shed a group B rotavirus detected by IEM. These findings provide additional evidence for a possible role for bovine coronavirus in the aetiology of winter dysentery. Furthermore, this is the first report of a group B rotavirus associated with diarrhoea in adult cattle.     

Epidemiological studies of piglet diarrhoea in intensively managed Danish sow herds. IV. Pathogenicity of porcine rotavirus

Colostrum-deprived piglets inoculated with rotavirus 24 h after birth developed a profuse diarrhoea that spread to non-inoculated, colostrum-deprived litter mates and, occassionally, to colostrum-fed piglets. Case fatality rates in these 3 categories of piglets were 63.2%, 35.7% and 8.3%, respectively. Surviving piglets recovered in 1-2 weeks, but shedded virus via the faeces for up to 3 weeks p.i. The D-xylose test revealed severe malabsorption, with extremely flat absorption curves for up to 3-4 weeks p.i. Malabsorption was more marked in piglets with a long-lasting faecal virus excretion than in piglets where virus disappeared from the faeces within 10 days p.i. Infected piglets (colostrum-fed and colostrum-deprived) had decreased weight gains and were 5 days older at a bodyweight of 25 kg than non-inoculated controls. It is concluded that rotavirus is probably of significance in diarrhoeal syndromes in suckling piglets, alone or in combination with E. coli or other pathogens.     

规模化奶牛场犊牛腹泻的病原检测与分析报告

为了调查某规模化奶牛场犊牛腹泻的发病原因,试验采用血清抗体检测、细菌分离培养、寄生虫卵镜检和病毒核酸检测方法对腹泻犊牛的血样和粪便进行了检查。结果表明,该奶牛场牛病毒性腹泻/黏膜病病毒抗体阳性率为65.00%,牛轮状病毒抗体阳性率25.00%,牛冠状病毒抗体阳性率15.00%;分离到1 株结肠弯曲杆菌和1 株空肠弯曲杆菌;所检腹泻犊牛粪便和血液样品中,4 份粪便样品和1 份血液样品检出牛冠状病毒,2 份粪便样品检出轮状病毒,1 份粪便样品和1 份血液样品检出牛病毒性腹泻/黏膜病病毒,同一份粪便样品中同时检出牛病毒性腹泻/黏膜病病毒和牛冠状病毒;通过镜检,球虫卵囊检出率77.78%,在1 份粪样中发现蛔虫卵,2 份粪样中发现其他线虫卵。该牛场流行性腹泻的原因可能为夏季高温潮湿引起犊牛免疫力下降导致的多病原体感染。     

Evaluation of ELISA and electron microscopy for the detection of coronavirus and rotavirus in bovine faeces

Enzyme-linked immunosorbent assay (ELISA) was compared with electron microscopy in the examination of faeces from experimental calves and showed 100 per cent agreement in the detection of 19 bovine coronavirus and 15 bovine rotavirus electron microscope positive samples. In a limited field survey of calf diarrhoea 75 selected faeces were examined independently by ELISA and electron microscopy and the agreement between the two tests was 95 per cent for coronavirus and 84 per cent for rotavirus. A further comparison was made with 74 samples submitted for routine diagnosis and this yielded agreements of 82 per cent (coronavirus) and 89 per cent (rotavirus). Factors contributing to discrepant results were examined and the relative advantages and disadvantages of the two tests for routine detection of these enteric viruses are discussed.     

Viral agents associated with outbreaks of diarrhea in turkey flocks in Quebec.

The relative importance of various enteric viruses associated with diarrhea of turkey poults was investigated by an evaluation of specimens received since 1982. Specimens originated from one to eight week old turkey poults, with mild to severe diarrhea, from 114 flocks in 42 commercial operations located in southern Quebec. The acute phase of enteritis occurred usually in poults between two and four weeks of age. Clarified intestinal contents were examined by direct electron microscopy and enzyme immunoassays. Enzyme-linked immunosorbent assays were performed with antisera to bovine rotavirus group antigen, avian reovirus types 1 to 5, and the prototype strain of the turkey enteric coronavirus. The presence of viruses could be demonstrated by electron microscopy in 55.3% of the specimens, and at least five different viruses were incriminated either alone or in combination. The coronavirus was by far the most common enteric virus with a prevalence of 47.5%. By enzyme-linked immunosorbent assay, rotavirus, reovirus and turkey coronavirus were detected in 14.5%, 18.1% and 61.4% of the specimens, respectively. By electron microscopy, 56.6% of these cases were positive for at least one virus.     

Detection of rotavirus and coronavirus shedding in two beef cow herds in Idaho

Fecal samples were taken at the time of pregnancy examinations and at parturition from two beef herds. They were also taken from sick calves at the onset of disease, and from 25% of the healthy calves at 15 days of age. All fecal samples were examined by electron microscopy for viruses.

Four cows in herd A were detected excreting coronavirus, one at the time of the pregnancy examinations and three at parturition. The first cow was removed from the herd and the others calved at the end of the season. There were no sick calves.

No cows in herd B were detected excreting virus at the time of pregnancy checks, but fourteen coronavirus and two rotavirus carrier cows were found at parturition. All but two calves sampled had large numbers of virus particles in their feces. Clinical illness was associated with dams shedding virus and with nightly low temperatures.

     

Research on antibodies against BHV-1, BHV-2, BHV-4, BVD-MD virus, bovine adenovirus A and B, rotavirus and coronavirus in cattle in western Zaire: complementary results

Two-hundred bovine sera from western Zaire were screened for antibodies to 8 viruses: BHV-1, BHV-2, BHV-4, BVD-MD virus, bovine adenovirus A and B, bovine rotavirus and bovine coronavirus. Positive sera were found to all these viruses. For animals whose origin was undoubted, the main features were the high prevalence of infections by rotavirus and BHV-4 and the low prevalence of infections by coronavirus and BVD-MD virus.     

Plasma enteroglucagon and neurotensin levels in gnotobiotic calves infected with enteropathogenic and non-enteropathogenic viruses

Five gnotobiotic calves were each infected with five viruses. Each calf was inoculated with coronavirus at seven days old, followed by astrovirus, Newbury agent, parainfluenzavirus type 3 and rotavirus at intervals of two weeks. Three of the viruses were enteropathogenic (bovine coronavirus, bovine calici-like virus and bovine rotavirus) and two were not (bovine astrovirus and parainfluenzavirus type 3). Plasma levels of the peptide hormones enteroglucagon and neurotensin and faecal output were measured daily and xylose absorption was studied before and after each infection. A close correlation was found between a rise in plasma enteroglucagon and neurotensin and infection with enteropathogenic viruses. The three enteropathogenic viruses caused increased daily faecal output, and elevated plasma levels of enteroglucagon and neurotensin, while the non-enteropathogens did not. The calici-like virus and rotavirus but not the coronavirus caused xylose malabsorption.     

Viruses and virus-like particles detected during examination of feces from calves and piglets with diarrhea

A total of 763 fecal or intestinal samples from diarrheic calves and piglets were examined for viral content by immunofluorescence, electron microscopy or cell culture. Routine fluorescent antibody and cultural tests detected rotavirus (n=126), coronavirus (n=80) and bovine viral diarrhea virus (n=13). Electron microscopy detected rotaviruses (n=24) and coronaviruses (n=17) not identified by standard fluorescent antibody tests. Other viruses detected by electron microscopy included Breda virus-like particles (n=49), astroviruses (n=1), caliciviruses (n=1), rhabdoviruses (n=1), parvoviruses (n=2), enteroviruses (n=3), togavirus-like particles (n=2), and “chained” particles (n=5). Mixtures of several of the viruses were detected in a number of fecal samples.

The survey emphasized the value of electron microscopy as a broad-spectrum diagnostic tool.

     

Prevalence of rotavirus and coronavirus antigens in the feces of normal cows.

The prevalence of rotavirus and coronavirus shedding by adult cows was investigated using capture enzyme-linked immunosorbent assays. Fecal samples from 121 cows in a single herd were tested for the presence of rotavirus and coronavirus, either free or complexed with immunoglobulin. Free rotavirus was not detected in any samples while rotavirus-immunoglobulin complexes were detected in 53 of 121 (44%) samples tested. In contrast, free coronavirus was detected in six (5%) samples and coronavirus-immunoglobulin complexes were detected in 85 (70%) of the samples tested. Thus it appears that subclinical infection of cows by either of these viruses is common, possibly providing a source for infection of the neonate. These assays may therefore provide important information regarding the epidemiology of enteric virus infections and suggest means of improving management to prevent epidemics of neonatal diarrhea.     

A reovirus-like agent (rotavirus) associated with diarrhoea in neonatal pigs

Reovirus-like particles were detected by electron microscopy in the faeces of pigs with diarrhoea. Attempts to adapt the virus to grow in pig kidney cell cultures were unsuccessful. By immunofluorescent staining techniques the pig virus was shown to be antigenically related to the calf rotavirus. Colostrum-deprived piglets were readily infected with the virus. Viral replication occurred in villous epithelial cells in the small intestines of infected pigs, and virus was detected in the faeces for at least 7 days post infection. All infected piglets developed diarrhoea whereas uninfected controls did not.     

Isolation and preliminary characterisation of some bovine enteric viruses

Fifty-eight faeces samples from calves and yearling cattle with diarrhoea were subject to a virus cultural study in monkey kidney cells and bovine embryonic kidney and lung cells. The properties of the 6 viruses isolated indicated that 5 were bovine enteroviruses belonging to 2 biologically different groups, and the remaining virus was probably a rotavirus (reo-like virus).     

Detection of bovine coronavirus and type A rotavirus in neonatal calf diarrhea and winter dysentery of cattle in Quebec: evaluation of three diagnostic methods.

The use of direct electron microscopy, enzyme-linked immunosorbent assay, and protein A-gold immunoelectron microscopy for the identification of bovine coronavirus and type A rotavirus were examined. Two hundred and forty-nine samples from diarrheic calves and winter dysenteric cattle from seven geographic areas in Quebec were examined for the presence of viruses by direct electron microscopy of negatively stained preparations. In addition, all the samples were analyzed by enzyme-linked immunosorbent assay, and a random selection of 47 samples were also analyzed by protein A-gold immunoelectron microscopy. Thirty-nine percent of samples examined by direct electron microscopy contained viral particles; bovine coronavirus and type A rotavirus were the most common viruses involved. Overall agreement between any two of the methods used compared favorably with results obtained by others using similar methods. The presence of coronavirus and rotavirus in fecal samples obtained from neonatal calves and the presence of coronavirus in samples from winter dysenteric adult cattle suggested their etiological roles in the respective diseases. Furthermore, results from protein A-gold immunoelectron microscopy of coronavirus-like particles implied that a different coronavirus or some other viruses might be involved in these diseases. Finally, the efficiency of direct electron microscopy, enzyme-linked immunosorbent assay and protein A-gold immunoelectron microscopy as diagnostic tools is discussed.     

Recent epidemiological status of canine viral enteric infections and Giardia infection in Japan.

Epidemiology of canine enteric infections was studied. Rectal swabs collected from 95 dogs presented at animal hospitals during a period from January to June of 2000 were examined for enteric pathogens, including viruses and Giardia lamblia (G. lamblia). Most frequently detected in both diarrheal and normal feces were canine coronavirus (55.4%) and G. lamblia (48.2%). Canine parvovirus type 2 (CPV-2) was specifically associated with diarrheal cases and CPV-2b was the predominant antigenic type. Although canine rotavirus, canine adenovirus, and canine distemper virus were also detected in a small number of diarrheal cases, no evidence for calicivirus infection was obtained.     

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.