Evaluation of a commercial ELISA test for the detection of a group A rota virus in pig faeces.
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Biomedical subjects
Publications and source records attributed to Z F Fu.
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Polyacrylamide gel electrophoresis (PAGE) and enzyme-linked immunosorbent assay (ELISA) were employed to investigate the epidemiology of typical and atypical rotavirus infections in five piggeries. Of 152 faecal samples examined, 46 (30 per cent) were positive by ELISA for group A rotavirus. Rotaviruses with electrophoretic patterns resembling groups A, B and C were detected. At least two and up to five different rotavirus electrophoretypes (typical and/or atypical) were detected in each of the five piggeries. Out of 152 faecal samples examined, 28 (18 per cent) contained rotaviruses with group A electrophoretypes, 9 (6 per cent) with group C but only 1 with Group B. Six samples contained both group A and group C rotaviruses. No common electrophoretypes of group A or C rotaviruses were detected in these five piggeries. The PAGE technique was also used to analyze group A rotavirus isolated sequentially from another piggery over a three year period. A single electrophoretype was found during the first two years, but in the third year a different electrophoretype was detected.
A severely dehydrated foal with a history of acute diarrhoea was presented to Massey University for treatment. Rotavirus was demonstrated in the faeces by electron microscopy. The failure to detect other pathogens suggested that rotavirus was the primary aetiological agent. Successful treatment of this case is described and the principles of treatment for diarrhoea, particularly fluid therapy, are reviewed. Antibodies to rotavirus were detected in the sera of nine mares and their foals on the stud that referred the case, indicating that rotaviral infection was probably endemic.
Five litters of piglets born within two days of each other, together with their dams, were investigated for faecal excretion of group A rotavirus antigen from birth to two months old. All the 50 piglets in these litters became infected with the virus between 19 and 35 days old. Rotavirus excretion was first seen in one litter which was housed with other litters not included in this study. Two days later, piglets of the second litter in another farrowing room began to excrete rotavirus, and then infection spread to the other three litters in the same room. Within each of these litters, one or two piglets were infected early and thereafter infection spread to other piglets. It took four to 10 days for rotavirus to infect every piglet within a litter, and 16 days in total before all piglets in the five litters were infected. No rotavirus antigen was demonstrated in faeces from sows during the investigation period.
In an experimental study on a piggery it was found that haemolytic Escherichia coli of O-serotypes 138 or 139 proliferated in the intestinal tracts of pigs following weaning, with E. coli of the O-138 type also being occasionally recovered from unweaned pigs, and once from a sow. Organisms of the O-138 type produced heat labile enterotoxin and their presence in weaned pigs was associated with the development of severe post-weaning diarrhoea. E. coli of O-139 type produced a vero cell cytotoxin and were associated with a milder diarrhoea in weaned pigs. Under various managemental circumstances the O-138 type E. coli almost invariably proliferated after weaning. The O-139 strain of E. coli did however proliferate rather than the O-138 strain following the movement of weaned pigs to new accommodation, after weaned pigs were returned to their sow and then weaning again 5 days later, and very occasionally in pigs weaned at 5 weeks of age. In all these cases earlier proliferation of the O-138 E. coli had been detected, suggesting that this may be a prerequisite for proliferation of the O-139 strain.
Attempts were made to induce an intestinal hypersensitivity response to weaner diet by feeding pigs with small quantities of this material before weaning. In two trials using different weaner diets piglets subjected to this regimen showed no significant differences in small intestinal structure, in ability to absorb xylose, in bodyweight gain, in incidence of diarrhoea or excretion of enteropathogens after weaning compared with pigs not given any of the diet before weaning, or fed with a different diet before weaning. When post-weaning diarrhoea occurred it was associated with an earlier, more prolonged and greater proliferation of enterotoxigenic Escherichia coli in the small intestines than occurred in healthy pigs after weaning. The greater proliferation in pigs which developed diarrhoea could not be attributed either to an excessive dietary intake after weaning, or to a specific proliferation of rotaviruses.
Attempts were made to discover the source of strains of haemolytic Escherichia coli infecting weaned pigs on a piggery. The organisms were not detected in the faeces of sows in the farrowing house, or in the in the faeces or intestinal tracts of slaughtered bacon pigs or sows. Sows held in a quarantine unit, and their offspring born in the unit, did not excrete haemolytic E. coli until after they were returned to the piggery. The environment of the piggery was the most likely source of infection for weaned pigs, and routine cleaning and disinfection of the accommodation did not prevent infection. Unweaned pigs were however able to transfer haemolytic E. coli to a newly built, previously unused weaning house, and establish a cycle of infection.
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Cross-sectional and cohort studies were conducted in a piggery to investigate the excretion pattern of group A rotavirus in pigs. The cross-sectional survey revealed that 47 (9 per cent) of 521 pigs sampled were excreting rotavirus in the faeces. No rotavirus antigen was detected in the faeces of pigs either less than one week or over two months old. The prevalence of infection increased with age over the sucking period, and was greatest at five weeks old. Diarrhoea was observed in only eight (17 per cent) of the pigs excreting rotavirus. Sixteen piglets from four litters were selected and faecal samples collected daily from each animal from birth to two months old. All the piglets excreted group A rotavirus and the range of ages at which they first became infected was between 13 and 39 days. The average duration of excretion in individual piglets was 7.4 days. Ten of 13 sucking piglets which excreted rotavirus developed diarrhoea soon after it was first detected.
An indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of antibodies against equine herpesvirus type 2 (EHV-2) in equine sera. The optimal conditions of antigen concentration, and serum and conjugate dilutions were established by chequerboard titrations. When the standard ELISA test was used for titration of test sera, it was found to give titres approximately 1500 times higher than those obtained in the virus neutralization (VN) test, and a correlation coefficient of 0.815 was obtained between these two tests on 42 equine sera. All the positive serum samples by the VN were also positive by the ELISA, and one negative serum in the former test was found to be positive in the latter. Under field conditions, the test also detected increases in antibody titres against EHV-2 in 13 out of 14 foals soon after these animals excreted the virus.
Viruses causing or associated with respiratory disease in horses worldwide are reviewed. Results are presented from a serological survey of 121 New Zealand foals and horses that had been affected by respiratory disease, determining the prevalence of antibodies in this country to the major viruses associated with similar disease overseas. To date there is no evidence of equine influenza virus in New Zealand. Both equine herpesvirus type 1 and 2 have been frequently isolated and show high serological prevalences. Serological evidence of equine rhinovirus type 1 and type 2 is presented with a prevalence of 12.3% and 41.2% respectively observed in foal sera, and 37.7% and 84.9% in adult horse sera. Antibody reacting to equine viral arteritis virus antigen was detected in 3/121 test sera. Equine adenovirus has been isolated on occasions and has shown a 39% serological prevalence in one study reviewed. Progress in New Zealand equine virus research is discussed.
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The epizootiology of equine herpesvirus type 2 (EHV-2) infection was investigated in Thoroughbred foals on a stud farm which in previous years had suffered economic loss due to respiratory disease. Sixteen pairs of foals and their dams were selected for this study and all of the foals became infected with EHV-2 by two to four months of age. These animals responded serologically to the virus infection as detected by an enzyme-linked immunosorbent assay (ELISA). EHV-2 infection persisted in these foals for two to six months with constant or intermittent virus recovery. This persistent infection stimulated continuous production of antibodies against EHV-2. As soon as the antibody levels reached their peak at five to six months, the isolation rate of EHV-2 from the nasal cavity of these animals decreased, and eventually by nine months of age virus could no longer be recovered. Respiratory disease was observed in ten of the 16 foals; and two severely affected animals died at two months of age. EHV-2 was isolated from both foals at ante and/or post mortem examination. It is postulated that EHV-2, either as an initiating agent or by means of immnunosuppression, caused the respiratory disease observed in these foals.
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PURPOSE: To examine the prevalence of myopia in rural and urban schoolchildren in Xiamen, China, and to assess the impact of environmental factors on rates of myopia. METHODS: Second-grade children attending either a city (n=119) or rural (n=91) school in Xiamen, China, were examined using cycloplegic autorefraction and biometry. Detailed questions on socioeconomic status, near-work activity, reading and writing habits, and family histories of myopia were asked in a face-to-face interview using a standard questionnaire. RESULTS: The prevalence of myopia was 19.3% (95% confidence interval [CI], 12.3, 29) in the city and 6.6% (95% CI, 2.4, 14.3) in the countryside. The average hours per day children spent reading and writing outside of school was 2.2 hours in the city compared with 1.6 hours in the countryside (P<.0001). In both schools, the odds ratio for total reading and writing, adjusted for parental history of myopia, was 2.2 (95% CI, 1.1, 4). CONCLUSION: These data suggest the prevalence of myopia is higher in the city than in the countryside. One possible explanation for these different rates could be that schoolchildren in the city spend more time reading and writing outside of school compared with children in the countryside. Myopic children in both the city and the countryside spent more time reading and writing compared with nonmyopic children. This increased near-work activity may contribute to the prevalence of myopia.