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J C Bridger

Publications and source records attributed to J C Bridger.

At least 55 records · Page 3Linked to original sources

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.

Animals↗

Antigenic and pathogenic relationships of three bovine rotaviruses and a porcine rotavirus.

The antigenic relationships between three bovine rotaviruses, UK, CP-1 and PP-1, and a porcine rotavirus were investigated; their pathogenicity for piglets was also assessed. After propagation and cloning in cell culture, the four viruses were used to produce convalescent and hyperimmune antisera in gnotobiotic animals. For the UK and PP-1 viruses, reciprocal virus neutralization tests gave ratios of homologous to heterologous titres in the range 57 to greater than 2300. Taking a 20-fold difference in titre to be the criterion of heterogeneity, these two viruses could be regarded as separate serotypes. The third bovine virus, CP-1, was identical to PP-1 in reciprocal neutralization tests but gave a one-way cross-reaction with UK convalescent sera, i.e. it appeared to be intermediate between the two serotypes. UK and CP-1 differed antigenically from the porcine virus as shown by homologous: heterologous neutralization titre ratios of 30 to 12 600. The bovine virus, PP-1, however, had a closer relationship with the porcine virus, the homologous: heterologous ratios ranging from 4 to 110. Oral inoculation of piglets with the bovine viruses revealed differences in their pathogenicity. Isolates UK and CP-1 caused no clinical disease and could not be passaged. However, on the first and second passages in pigs, the bovine virus PP-1 and the porcine virus produced clinical disease, excretion of virus in the faeces and sero-conversion. The results suggest that PP-1 may be a natural hybrid between a bovine and a porcine rotavirus.

Animals↗

Characterization of a calici-like virus (Newbury agent) found in association with astrovirus in bovine diarrhea.

A bovine calici-like virus and astrovirus, present in the same fecal sample from an outbreak of diarrhea, were separated from each other by calf passage. The calici-like virus (Newbury agent SRV-1) caused anorexia, diarrhea, and xylose malabsorption in gnotobiotic calves aged 17 to 60 days, whereas the bovine astrovirus was nonpathogenic in similar calves. The calici-like virus was shown to be antigenically distinct from a previously described isolate (Newbury agent SRV-2) by two-way cross-protection experiments in calves; calves immune to homologous challenge became clinically ill and excreted virus when challenged with the heterologous virus.

Animals↗

Localisation of enteropathogens in paraffin embedded tissue by immunoperoxidase.

An indirect immunoperoxidase technique has been used to identify enteropathogens in formol-sublimate fixed paraffin embedded sections of calf intestine. Infections with bovine rotavirus, bovine coronavirus, Newbury agent SRV -1, and K99+ Escherichia coli have been detected in the intestines from experimentally infected and conventially reared diarrhoeic or normal calves. The ability to visualize enteropathogenic agents in histological sections resulted in the demonstration of virus infected cells at sites not previously shown to be infected using the immunofluorescence technique.

Animals↗

Lesions of gnotobiotic calves experimentally infected with a calicivirus-like (Newbury) agent.

Fourteen gnotobiotic calves were killed 0.5 to ten days after infection with Newbury agent SRV-1 and the changes in small intestinal structure and function were assessed, qualitatively and quantitatively, by light microscopy, scanning and transmission electron microscopy, enzymology and xylose absorption. The first enterocytes detected as infected by immunoperoxidase were those on the sides of villi at the base. Subsequently exfoliation of degenerate enterocytes resulted in stunted villi; mucosal beta-galactosidase activity fell and there was xylose malabsorption. Small intestinal damage, first detected at 12 hours after infection but almost repaired by ten days, was restricted to the anterior half of the small intestine. In the distal small intestine, where no virus-induced damage occurred, villi lengthened--possibly due to increased mitosis of crypt cells stimulated by enteroglucagon release.

Animals↗

Molecular characterization of rotaviruses with distinct group antigens.

Comparative serological and nucleic acid studies were carried out on rotavirus isolates, with and without the original group antigen. Indirect immunofluorescence allowed the atypical isolates to be divided into two groups each with its own distinct group antigen hence giving a total of at least three distinct rotavirus groups. Genome profile analysis of the atypical isolates again divided them into two groups which corresponded with those obtained by immunofluorescence and within which the overall pattern of genome segments was similar. A more rigorous examination of the degree of difference between the three virus groups was carried out using single-dimension terminal fingerprint analysis. This indicated that the viruses in the three groups did not carry any genome segments in common with each other. Therefore, experimental criteria have been established which allow the division of rotaviruses into three groups. Analysis of further atypical isolates, as they become available, will establish whether the division of rotaviruses into a number of separate groups, of the type seen in influenza virus, is justified.

Animals↗

Characterization of an antigenically distinct porcine rotavirus.

A porcine virus with rotavirus morphology, which was antigenically unrelated to previously described rotaviruses, is described. Particles with an outer capsid layer measured 75 nm and those lacking the outer layer were 63 nm in diameter. Particles which resembled cores were also identified. The virus was shown to be antigenically distinct from other rotaviruses as judged by immunofluorescence and immune electron microscopy, and it failed to protect piglets from challenge with porcine rotavirus. Analysis of the genome indicated that it was composed of 11 segments of double-stranded RNA with the same overall size range as other rotaviruses but with an unusual segment pattern.

Animals↗

A search for new microorganisms in calf pneumonia by the inoculation of gnotobiotic calves.

In a detailed microbiological and pathological study of eight outbreaks of calf pneumonia, which included the inoculation of respiratory material into 18 gnotobiotic calves, a total of five viruses, four species of mycoplasma and 19 species of bacteria were identified. The only microorganism not previously associated with bovine respiratory disease was a coronavirus. The experimental disease produced in the majority of the gnotobiotic calves by the intratracheal inoculation of unpassaged respiratory material, closely resembled the natural disease. However statistical analysis of the results could not ascribe the disease seen to a single microorganism or to a particular combination of microorganisms. Assuming therefore that no microorganisms were missed in the study, it was concluded that the microbiological cause of the experimental disease lay amongst the organisms already found and that one or more of three other factors, discussed in the paper, accounted for the experimental disease produced.

Animals↗

Development of immunity to porcine rotavirus in piglets protected from disease by bovine colostrum.

Bovine colostrum with rotavirus-neutralizing activity was fed for 10 days to two groups of piglets, one of which was inoculated intranasally with a rotavirus of porcine origin. A third group, which did not receive colostrum, was also inoculated with the virus, and these piglets developed diarrhea, excreted rotavirus in the feces, and died 6 days after infection. In contrast, the infected piglets fed with bovine colostrum remained healthy, although they developed antibody to rotavirus. Twenty-seven days after the primary inoculation, piglets in the colostrum-fed groups were inoculated intranasally with virus. Those in the previously unexposed group became clinically ill and excreted rotavirus, whereas those which had experienced a previous subclinical infection (the colostrum-fed, virus-inoculated group) remained healthy. It was concluded that bovine colostrum protected piglets from the clinical effects of a porcine rotavirus and that these animals developed an immunity which prevented subsequent disease.

Animals↗

Replication of an enteric bovine coronavirus in intestinal organ cultures.

A coronavirus isolated in tracheal organ cultures from the faeces of a calf with diarrhoaea readily multiplied on passage in intestinal organ cultures. Evidence for multiplication was obtained by the production of viral haemagglutinin in organ culture fluids and the presence of immunofluorescence and viral particles in the columnar epithelial cells of the villi. Thus virus multiplication was studied in vitro in the cell type in which it multiplies naturally.

Animals↗

Isolation of small viruses resembling astroviruses and caliciviruses from acute enteritis of calves.

Small round viruses (SRV) were isolated from the faeces of diarrhoeic calves from three farms. All three SRV preparations caused diarrhoea experimentally in gnotobiotic calves. Each preparation contained viral particles of two morphological types, "astrovirus-like" and "calicivirus-like", and from one preparation the two particle types were separated from each other. The calicivirus-like agent ("Newbury agent") was 33 nm in diameter, and caused diarrhoea in gnotobiotic calves with villous atrophy and D-xylose malabsorption. This virus did not infect cell cultures. The astrovirus-like agent did not cause diarrhoea in two gnotobiotic calves; however, it infected cell cultures (primary calf kidney) and the infected cells immunofluoresced with convalescent gnotobiotic-calf antiserum. The astrovirus-like agents in the three preparations were antigenically related. Experiments in calves showed that there was a degree of cross-protection between the three SRV preparations, as judged by the presence or absence of diarrhoea, but that at least three unrelated pathogens were present.

Acute Disease↗

Location of type-specific antigens in calf rotaviruses.

Rotavirus antisera with and without neutralizing activity to calf rotavirus were compared for their ability to agglutinate calf rotavirus particles with and without an outer capsid layer. Particles without the outer capsid layer were agglutinated by antisera with immunofluorescent antibody activity and by antisera with immunofluorescent antibody and neutralizing activity. Particles with the outer capsid layer were agglutinated only by antisera with neutralizing activity. The neutralization test appears to be type specific, and it was concluded that type-specific antigens are associated with the outer capsid layer of rotaviruses. Since particles with the outer capsid layer were not agglutinated by antisera lacking neutralizing activity but possessing immunofluorescent antibody activity, it was concluded that in intact particles the group-specific antigen is masked by the outer capsid layer.

Animals↗

Serological relationships between rotaviruses from different species as studied by complement fixation and neutralization.

Human, piglet, mouse, foal, lamb, calf and rabbit rotaviruses all infected, but could not readily be subcultured in LLC MK2 cells. Cells infected with mouse and calf rotaviruses reacted by indirect immunofluorescence (FA) with convalescent serum from children, piglets, mice, foals, lambs, calves or rabbits, taken after rotavirus infection. Human, calf, piglet, mouse and foal rotaviruses reacted with human, calf, mouse, foal and lamb convalescent serum by complement fixation (CF). It was not possible to distinguish between different rotaviruses by CF or FA. Neutralization tests, however, detected species-specific rotavirus antigens. Any virus was neutralized by a much higher dilution of homologous species convalescent serum than by any heterologous serum. With the exception of the mouse virus there was very little cross reaction. However, in sera with a very high neutralizing titre for the homologous virus the titre was proportionately raised against heterologous virus. It is, therefore, now possible to type to species an unknown rotavirus by a neutralization test in LLC MK2 cells using convalescent serum from each species.

Animals↗