Michael Clarkson

The turkey, compared to the chicken, fails to mount an effective early immune response to Histomonas meleagridis in the gut.

Histomonosis is a disease of poultry caused by Histomonas meleagridis. Chickens usually recover while the mortality rate in turkeys is high. The immunological response of both species towards H. meleagridis was investigated. Parasites migrated in greater numbers to the turkey liver compared with that of chicken. Chicken mounted an effective caecal innate response, with increased expression of IL‐1β, CXCLi2 and IL‐6 mRNA, resulting in control of parasite numbers. The turkey failed to mount such an effective innate response in the caecal tonsil, allowing greater numbers to migrate to the liver, where a sustained, uncontrolled immune response was mounted, evidenced by the upregulation of mRNA for IL‐1β, CXCLi2, IFN‐γ, IL‐13, IL‐4 and IL‐10. Expression levels of mRNA of the chicken and turkey β‐defensin AvBD2 suggest that this response was not limited to the cytokines. There was an influx of CD4+, CD8α+, CD28+ and CD44+ cells into the livers of both species, coinciding with parasite movement. These influxes were more pronounced in the turkey, correlating with a decrease in numbers of the same cells in the spleen, which was not observed in the chicken. Antibody levels in the chicken increased more than those in the turkey, supporting evidence of an adaptive response.

Analyses of the genomes of chlamydial isolates from ruminants and pigs support the adoption of the new species Chlamydia pecorum.

Analysis of the genomic DNAs of chlamydial isolates from sheep, cattle, and pigs was performed by Southern blot hybridization and by restriction endonuclease (RE) profiling of DNA amplified by PCR. The hybridization probes were derived from whole genomic DNA, the major outer membrane protein (MOMP) gene, the 16S rRNA gene, and an avian Chlamydia psittaci isolate plasmid. The PCR analysis used targets in the MOMP gene, the 16S rRNA gene, and the 60-kDa cysteine-rich protein gene. Together, the results showed that although there was considerable heterogeneity in the DNA sequence in the MOMP gene region, all the isolates had the same underlying total genomic RE profiles and yielded identical RE profiles for the rRNA and 60-kDa-protein gene regions. Most of the isolates were found to hybridize with the plasmid probe. Comparison of the MOMP sequence of one of the isolates (P787) with that of a known Chlamydia pecorum strain together with the results of the RE analyses allowed the conclusion that the isolates should all be classified within this new species.

Validating indicators of sheep welfare through a consensus of expert opinion

A consensus of expert opinion was used to provide both face and consensual validity to a list of potential indicators of sheep welfare. This approach was used as a first step in the identification of valid welfare indicators for sheep. The consensus methodology of the National Institute of Health, using pre-meeting consultation and focus group discussions, was used to ascertain the consensus opinion of a panel of sheep welfare experts. The Farm Animal Welfare Councils five freedoms were used as a framework to organise a list of current on-farm welfare issues for sheep. The five freedoms were also the welfare criterion used to identify potential on-farm welfare indicators for sheep. As a result, experts identified 193 welfare issues for sheep and lambs managed on farms across England and Wales. Subsequently, a combination of animal- (n = 26), resource- (n = 13) and management- (n = 22) based indicators was suggested for (i) adult rams, (ii) adult ewes (male and female sheep, over 1 year old), (iii) growing lambs (male and female sheep, over 6 weeks to 1 year old) and (iv) young lambs (male and female lambs, 6 weeks old and under). The results from this study could therefore be used to inform the further development of valid methods of assessing the on-farm welfare of sheep.

The epidemiology of fasciolosis in the inter-Andean valley of Cajamarca, Peru

Fasciolosis is recognised as a major problem in dairy cattle in parts of Peru. A longitudinal study of dairy cattle in Cajamarca, Peru was used to determine the annual pattern of infection with Fasciola hepatica. After a gradual increase from January, peak egg production occurred in August/September and then dropped rapidly. Indirect indicators of infection, eosinophil counts and serum liver-enzyme activities, indicated that the major period of new infection in cattle occurred from December to May each year. Examination of snails demonstrated that, although there was no clear annual cycle in snail abundance, the majority of snails infected with cercariae were found between January and March. Climatological data indicated that there was sufficient moisture for development of the parasite during a limited period each year, coinciding with the period of maximum abundance of cercaria-infected snails, but that irrigation could substantially alter the amount of water available. Infections in tracer-calves decreased from December to September, with little or no infection occurring between June and August, but suggesting that there could be significant infection prior to December. Thus a defined annual cycle of infection was observed, where cattle acquired infection from December to May and this infection matured to produce peak egg counts in August/September which were then available to infect the intermediate host for the next cycle of infection.

Antigenic and morphological differentiation of placental and intestinal isolates of Chlamydia psittaci of ovine origin.

Ewe placental and lamb intestinal isolates of Chlamydia psittaci recovered from flocks affected with ovine enzootic abortion were examined by inclusion morphology, indirect immunofluorescence (IIF) and immunoblot analysis. Chlamydiae recovered from the faeces of sheep from two flocks free of clinical disease were also examined. In cell culture ovine abortion (OA) and intestinal isolates were distinguishable by inclusion development and morphology. Similarly, in two-way IIF tests with one week mouse antisera isolates fell into two distinct groups: abortion or intestinal. Immunoblotting with convalescent sheep abortion antiserum identified 30 out of at least 40 silver staining polypeptides as antigenic both in OA and intestinal isolates. The serum produced a similar reaction pattern to the resolved proteins of each OA isolate, indicating a higher degree of antigenic conservation among these isolates. Considerable cross reactivity between the OA and intestinal isolates was identified, but the serum also showed apparent molecular weight differences between antigens of the two types in the 87-116 kDa, 38-44 kDa and 26-28 kDa regions. Furthermore, the immunoblotting analysis revealed heterogeneity among the intestinal isolates, particularly in antigens between 87-116 kDa and 38-44 kDa.

Bovine immune recognition of Ostertagia ostertagi larval antigens.

Analysis of a detergent-solubilized somatic antigen of Ostertagia ostertagi 3rd stage larvae by SDS-PAGE and Western blotting has revealed two specific antigens with apparent molecular weights of 17 and 43 kD under reducing conditions. Probing of the Ostertagia ostertagi preparation with preinfection control sera has shown two cross-reacting antigens with apparent molecular weights of 67 and 81 kD. Both the 17 and the 43 kD antigens were recognised by IgG1 which was the predominant reactive subclass. FPLC fractionation of the crude extract with gel filtration and ion-exchange columns demonstrated immune reactivity in discrete peaks. Comparisons of ELISA and lymphocyte transformation showed antigenic components reactive with both antibodies and primed lymphocytes.

Isolation of faecal chlamydia from sheep in Britain andtheir characterization by cultural properties

Faeces samples were taken per rectum from sheep on 26 farms in England and Wales and examined for the presence of chlamydia by culture in McCoy cell monolayers. Thirteen of the farms were known to have had abortion outbreaks associated with Chlamydia psittaci (enzootic abortion) and 13 were free of this infection. The chlamydia isolated were characterized by cultural techniques. Chlamydia were isolated from the faeces of lambs on all 26 farms and the prevalence of infection varied form 5-50% on individual farms. There was no significant difference between the proportion of infected lambs on farms where enzootic abortion was present or absent. Lambs first showed infection when they were 3 months old and the prevalence rate of infection increased up to 9 months old. No chlamydia were isolated from the faeces of 316 adult ewes. The chlamydia were identified as enteric rather than abortion type and thus were C. pecorum rather than C. psittaci.

The specificity of antibody responses in cattle naturally exposed to Fasciola hepatica.

Fasciola hepatica causes significant morbidity and mortality in dairy cattle in the Andean region of Cajamarca, Peru, where prevalence of infection of up to 78% has been reported. ELISA and Western blot analyses were used to characterise antibody responses in dairy cattle to adult F. hepatica to excretory-secretory (E/S), somatic (SO) and surface (SU) antigens. Three groups of dairy cattle - calves, heifers and adult cows - naturally exposed to F. hepatica in this region, were monitored every 2 months over a 2-year period. Calves, heifers and adult cows all had antibodies which recognised a 28kDa protein in the SO preparation, whereas only adult cows had antibodies that recognised a 28kDa protein in E/S products. All three groups of cattle responded to a 60-66kDa group of proteins in E/S and SU preparations and a 17kDa antigen in SO products was recognised by antibodies from cows and heifers but not calves. The total antibody response to E/S antigens measured by ELISA, increased over time in calves and remained constantly high over the 2-year period in all three groups of cattle. Slight fluctuations in the antibody response occurred in the group of heifers and cows coinciding with seasonal changes in the level of challenge.

Comparison of Albendazole and Praziquantel therapy of Echinococcus granulosus in naturally infected sheep.

The effects of albendazole (10 mg kg-1 day-1) and praziquantel (50 mg kg-1 day-1) for 6 weeks on naturally infected sheep with pulmonary cysts of Echinococcus granulosus of proven viability were studied. Immediately following therapy, one of three sheep treated with praziquantel had viable cysts and 7 months later one of two sheep had viable cysts. One sheep died during albendazole therapy, but 7 months following therapy only one of five sheep had viable cysts. Electron microscopy demonstrated necrotic germinal layer tissue in most albendazole-treated cysts and praziquantel also had an effect on cyst ultrastructure. These data suggest that recurrence in humans treated with albendazole may be small. Whilst praziquantel was not particularly effective in this animal model, its clear effect on the ultrastructure suggests that an increased dose and combination therapy with albendazole may be more effective.

The prevalence of Echinococcus granulosus in farm dogs in mid-Wales

Abstract One hundred and fourteen farms in Powys, mid-Wales, were visited on two occasions approximately 8 months apart during 1975. All the farm dogs were dosed with arecoline acetarsol and the faecal purges were examined for the presence of Echinococcus granulosus . Sixty-seven farms (58.8%) contained at least one infected dog on at least one of the visits but only 36 farms and 43 farms had infected dog at the first and second visits, respectively. Four hundred and twenty-nine dogs provided at least one sample and 108 (25.2%) of these samples contained E. granulosus . Only seven dogs were infected at both visits. The mean numbers of worms found in the samples were 241.6 and 156.4 at the first and second visits, respectively, but almost 50% of the positive samples contained fewer than 26 worms. The worms were identified as the sheep strain, the known cause of hydatid disease in man in Wales.