The development of new and effective vaccines and immunodiagnostic reagents requires the characterisation of antigenically relevant proteins and their interactions with the products of the immune system. Phage display technology was investigated as a means of elucidating some of the antigenic properties of the rickettsial parasite, Cowdria ruminantium (Cowdria). Randomly fragmented gene-derived libraries have been useful in elucidating viral and other epitopes, but only limited work has been done with entire genomes. A phage display library expressing a repertoire of Cowdria peptides was constructed. It was sufficiently large to represent the organism's genome, but lacked phages displaying peptides coded for by genes containing a Pvu II restriction enzyme site, including the one coding for the major antigenic protein 1 (MAP1). This was considered advantageous since MAP1 is immunodominant and has already been well characterised. Affinity selection with antibodies against Cowdria proteins other than MAP1 allowed several antibody-reactive peptides to be isolated. These selected sequences were placed in the context of the genome by screening a lambda bacteriophage library and by comparison with Cowdria DNA sequences. Apart from showing that antigenic mimics were present in the phage display library, six open reading frames encoding putative Cowdria proteins were identified. All had similarities to, or motifs in common with, membrane proteins and are thus likely to be exposed to the host's humoral immune system. Some of the proteins identified were larger than the antigens used to elicit the antibodies used for selection, probably as a result of the presence of cross-reactive epitopes. Despite limitations experienced when extending a fragmented-gene approach for epitope location to genomes, it was possible to identify an antigenic region on MAP1 by comparison with selected mimics. In addition, binding peptide sequences were identified with two monoclonal antibodies that had been raised against non-Cowdria antigens. An epitope on the VP7 protein of bluetongue virus was identified and peptides were found that reacted with a monoclonal antibody directed against malignant catarrhal fever virus. Thus, apart from being able to identify several potentially important Cowdria epitopes and genes, the fragmented-genome library holds promise as a universal reagent for identifying useful mimics.

Call number: LD2668 .T4 1978 C38 / Master of Science

Call number: LD2668 .T4 1978 M78 / Master of Science

No description available.

Nematode parasites have become the biggest problem for small ruminant production in South Africa due to their resistance to the commercial anthelmintics. Notable, wattle tannin has been used as an alternative strategy for control. However, the concentration and the frequencies can likely influence its effect on the parasites control. The objective of this study was to determine the degree of pasture infestation and nematode infection in sheep and goats, as well as investigate nematode resistance to the anthelmintics, and the potential of wattle tannin in nematode control. The first study dealt with the epidemiology of internal parasites. Eight Merino ewes and eight Nguni does averaged 7-18 months of age were observed for 1 year during the months of February 2008 to January 2009 at the University of KwaZulu-Natal Research Farm (Ukulinga). Egg count per gram (EPG) and coccidian oocysts per gram (OPG) were counted according to Mc Master Technique (Hansen & Perry, 1994) by magnifying parasitic eggs from monthly rectal faecal samples dissolved in saturated sodium chloride. Faecal samples also were cultured for 15 days to identify infective nematode larvae (L3) using Baermann technique. Herbage samples were collected monthly from four paddocks as well to count L3 on the pasture. Sheep live weight was also recorded monthly. Seasonal effects was significant (P<0.05) on the EPG, OPG, faecal culture L3 and pasture L3. A higher level of infection was observed in summer (wet) than in winter (dry season). Trichostrongylus spp larvae were the most prevalent larvae (26.5%) while Strongyloides, Haemonchus contortus, Nematodirus and Cooperia spp occurred in the faecal culture by percentage of 20.9%, 16%, 16% and 14.5%, respectively. For parasite resistance, Ivermectin 1% (IVM), Closantel 5% (CST) and a combination of Abamectin 0.08% and Praziquantel 1.5% (CPA) were evaluated. Twenty four sheep (12 females and 12 males) aged between 7-18 months were used for 21 days. Animals were naturally infested by gastro-intestinal parasites. EPG and faecal culture L3 were counted on day 0, 7, 14 and 21. Closantel was the most effective. Haemonchus spp. were least affected whilst Trichostrongylus spp. were the most affected by all drugs. In the third study, wattle tannins were evaluated as an alternative nematode control drug. Three experiments (Exp.) were conducted to determine the effect of tannin concentration (Exp.1 and 2) and frequency (Exp.3) on nematode parasites. In Exp.1, 0, 0.8, 1.6 and 2.4 g tannin/kg BW were drenched for three consecutive days per sheep (16 females and 8 males, aged 8-9 months) for 21 day. In Exp.2, 30 sheep (14 males and 16 females, aged 9-18 months) were randomly allocated into three tannin treatments (0, 0.8 and 1.6 g tannin/kg BW) and drenched for a day. In Exp.3, 26 sheep (11 males and 15 females aged 9-18 months) were divided into three groups of 9, 9, and 8 sheep each. These groups were drenched with 1.6 g tannins/kg BW/day; once, twice or thrice for the 3 groups respectively. For the three experiments, EPG and L3 larvae were counted in individual feacal samples. For all tannin treatments, EPG decreased (P<0.05) over time. Though the differences among tannin levels and frequencies varied (P<0.05) over time, EPG consistently decreased with increasing tannin level and frequency. Thus 1.6 and 2.4 g tannin /kg BW for 3 consecutive days had nearly similar effects on the EPG. The results of this study are rather inconclusive that weather conditions such as rainfall had a direct effect on internal parasites development. Nematode parasites at Ukulinga Research Farm were resistant to the commercial anthelmintics used. Drenching with 1.6g wattle tannin/kg BW over three successive days is enough to reduce EPG and reduce the degree of pasture contamination. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.

No description available.

Five crossbred heifers were slaughtered at different times and rumen and omasum tissues were removed: The epithelium was separated from the muscle layer and mounted in parabiotic chambers. The tissues were incubated for 2 h in buffer at 39 C and aerated continuously in 95% O₂ 5% CO₂ . The buffers were similar to those which have been used previously in mineral transport studies, but were modified by varying the levels of Mg and K while keeping the osmolarity constant. Potassium was included in the buffers on the mucosal side in appropriate ratios to Mg to correspond to the K: Mg in diets fed to ruminants which would contain .1% Mg and .6, 2.4 and 4.8% K, (low, medium and high, respectively). A fourth buffer which contained physiological concentrations of Kand Mg, was placed on the serosal side of the parabiotic chamber. Flow of Mg was calculated by: F = C₁V₁ - C₀V₀ / AΔt where C₁ = final concentration, C₀ = original concentration, V₁ = final volume, V₀ = original volume, Δt = incubation time, A = area of the tissue exposed to the buffer, and F = flow of Mg (mg/cm²/h). Blanks were included which contained physiological levels of Mg on both side of the chamber to adjust for tissue effects. Magnesium transport tended to be 10 times greater through the rumen than the omasum. This indicates that the rumen is the primary site of Mg absorption in cattle. Potassium tended to depress Mg transport across both tissues. This technique has only limited application in mineral research. / M.S.

Gastrointestinal nematode parasites cause great losses in the production of small ruminants through reduced productivity and the cost of preventive and curative treatments. Because of the threat of anthelmintic resistance, biological control of sheep nematodes has been identified as an alternative to anthelmintic drugs. Bacillus thuringiensis (Bt) (Berliner) and Clonostachys rosea (Schroers) have been widely studied as biocontrol agents. B. thuringiensis has been used for the biocontrol of insects and C. rosea has been successfully used as biocontrol agent of Botrytis cinera (De Bary) in plants. B. thuringiensis and C. rosea strains were isolated from soil collected from the Livestock Section at Ukulinga Research Farm, University of KwaZulu Natal, Pietermaritzburg. Twenty-five strains of Bt and 10 strains of C. rosea were successfully isolated. The Bt colonies were identified by their circular, white, flat and undulate character, and the gram-positive and rod-shaped endospores. C. rosea was identified by white colonies on Potato-dextose agar and the characteristic conidiophores, which were branched and showed phialides at the tips. In vitro screening of the isolates was undertaken to select the best isolates. The isolates that caused significantly greater mortality were Bt isolate B2, B10 and B12 and C. rosea isolates P1, P3 and P8. These isolates caused substantial nematode mortality in both faeces and water bioassay. Nematode counts were reduced by 28.5% to 62% and 44% to 69.9% in faecal bioassay for Bt and C. rosea, respectively. In the water bioassay, nematode counts were reduced by 62% to 85% for Bt and by 62.7% to 89.3% for C. rosea. The best inoculum level at which the best isolates were most effective, and the optimum frequency of application were determined. The trial was conducted using bioassays with faeces and water. Inoculum levels of 10(6), 10(8), 10(10), 10(12) spores ml-1 for Bt and 10(6), 10(8) and 10(10) conidia ml-1 for C. rosea was used in the faecal bioassay. The inoculum levels tested in water bioassay were 10(6), 10(8), 10(10) and 10(12) spores ml-1 for Bt and 10(9), 10(10), 10(11), 10(12) conidia ml-1 for C. rosea. In the faecal bioassay, B2 was the most effective Bt isolate at an inoculum level of 10(10) spores ml-1. Isolate P3 was the best C. rosea isolate at 10(8) conidia ml-1. In the water bioassay, Isolate P3 caused a mortality of 85% at inoculum levels of 10(9), 10(10) and 10(11) conidia ml-1. The performance of biological control agents in the field is sometimes inconsistent. Combining different biocontrol agents may be a method of improving their reliability and performance. However, the combination of most of the isolates was antagonistic, with efficacy less than that of either individual biocontrol agent. In particular, Isolate P3 was more effective when used alone than when combined with any other isolates. Therefore, the combination of biocontrol agents does not always result in synergistic interaction. There were some additive interactions between two bacterial isolates, and with one bacterial and fungal combination. The effect of feeding the best of the biocontrol agents, or diatomaceous earth (DE), was evaluated in sheep. Two doses of Bt (1g and 2g kg-1BW) and C. rosea (1g kg-1BW) reduced the numbers of L3 nematode larvae in sheep faeces. The DE product (at 15% of feed) also reduced L3 numbers but it was less effective than either the Bt or the C. rosea products. Nematode counts were reduced by 74.6%, 75.1%, 84.6%, 68.5% and 27.5% for Bt 1g kg-1BW, Bt 2g kg-1BW, C. rosea (1g kg-1 BW), DE and control, respectively. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2011.