SummaryThe strong association between polymorphisms in an intronic microsatellite and the coding sequences for (BoLA)-/)/^5i genes, previously described for demonstrating alleles of class II major histocompatibility complex (MHC) in the cow, was examined in sheep to see if similar polymorphisms could be demonstrated in the DRB region of the MHC. The bovine primers LA53 and LA54, previously used to amplify the bovine DRB3 microsatellites. were used with DNA from Australian sheep, eight DRB alleles were identified by length polymorphisms of poiymerase chain reaction (PCR) products amplified from the DRB microsatellite region. Incomplete amplification of both aileles was sometimes found for sheep DNA samples using bovine primers, so a modified primer (LA53b) was used, and found to amplify the microsatellite next to intron 2 of the MHC more reliably than the LA53 primer. Two additional primers {LA31 and LA32), used in amplification of the exon 2 region of bovine DRB3, were used in the sheep, and the PCR products were analysed by single-stranded conformation polymorphism (SSCP). These primers successfully amplified the variable region of the ovine DRB region coded by exon 2, and the SSCP technique demonstrated polymorphisms with sheep DNA. Family studies demonstrated the segregation of alleles, by amplification both of intronic microsatellites and of the exon 2 variable region. Close correspondence was found between the two regions for several alleles. suggesting that the intronic microsatellites were closely linked to DRB-\av\ab\e region alleles. Three families of Merino sheep with ditferent antibody responses to intestinal nematode parasites were examined. The sire group with the highest antibody levels possessed two mierosatellite alleles of closely similar length (alleles 3 and 4) inherited from the sire and present in high frequency in the lambs. In contrast, the other two sires did not possess these two alleles and the alleles were in low frequency in their progeny. Further studies are required in unrelated sheep to confirm whether these two alleles are associated with resistance to nematode parasites.
The hypothesis that behaviour in sheep is influenced by resistance to infections with Trichostrongylus colubriformis and Haemonchus contortus was explored. Sheep were assessed phenotypically as resistant and susceptible in four ways and thereafter, the effect of resistance on several behavioural traits was measured in an arena test. The behavioural parameters recorded for each sheep were: approach/avoidance distance; travel; number of moves; and spread. Four phenotypic groups of sheep were set up each with two subgroups: ovine lymphocyte antigen (OLA) type (subgroups SY1a type vs other types); blood eosinophil leucocyte counts (high vs low); T. colubriformis and H. contortus serum antibodies (high vs low titres) and faecal egg count (FEC) (high vs low). Only the behavioural comparison between sheep with high versus low eosinophil leucocyte count showed consistent differences that were statistically significant, although other comparisons favoured associations between OLA type SY1a, low FEC and reduced approach/avoidance distance and/or locomotor behaviour. The eosinophil leucocyte association was established on two non-overlapping test populations of sheep, with mean eosinophil leucocyte counts of 3.19 x 10(5)/mL blood for high and 0.83 x 10(5)/mL blood for low subgroups; results were reproduced on five separate occasions over 6 weeks to detect differences as approach/avoidance distance (P < 0.01), travel (P < 0.01), number of moves (P < 0.01), and spread (P < 0.05). High eosinophilia correlated with low FEC for the infected sheep, which identified sheep that were resistant to parasites. We concluded, therefore, that the resistant sheep were more at ease with the environment of the test than were the sheep with low eosinophil leucocyte counts. The strong association between high eosinophil leucocyte counts and behaviour is consistent with other reports which suggest that retention of normal levels of circulating eosinophil leucocytes is associated with resistance to stress.
The effect of dietary protein supplementation on the pathophysiology of a single Haemonchus contortus infection in 2-year-old Boer dry does under confined conditions was examined. Twenty-four does were randomly confined in individual pens and allocated into three groups balanced by their liveweight. The goats in the Control group were offered a basal diet whereas goats in Treatments 1 and 2 were fed the basal diet plus 25% and 50% dietary protein increments, respectively. Each animal was orally administrated 100 H. contortus L3 larvae/kg liveweight on Day 0 of the experiment. The goat liveweights and FAMACHA scores were recorded at weekly intervals. Packed cell volumes, eosinophil percentage, haemoglobin concentration, total serum protein, globulin, and albumin concentrations were recorded on Days 0 and 14, and then at weekly intervals during the experiments. Faecal egg counts were measured on Days 0, 21, and then at weekly intervals thereafter. Antigen-specific IgA, IgG, and IgM titres were determined from Enzyme-linked immunosorbent assays tests on Days 0, 28, and at the termination of the experiment. The results of this study showed that a single H. contortus L3 infection did not cause pathogenic effects in 2-year-old Boer dry does. Dietary protein supplementation significantly lowered faecal egg count and significantly enhanced IgG titres (P < 0.05). However, the higher protein diets did not have any significant effects on liveweight, packed cell volumes, haemoglobin concentration, biological parameters (total serum protein, albumin, and globulin concentrations), and peripheral antibodies (IgA and IgM titres) between the treatments. Further investigations are required to clarify the role of protein supplementation to control H. contortus infection in goats.
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