Scielo RSS <![CDATA[Onderstepoort Journal of Veterinary Research]]> vol. 80 num. 1 lang. pt <![CDATA[SciELO Logo]]> <![CDATA[<b>Construction of an artificial recombinant bicistronic plasmid DNA vaccine against porcine rotavirus</b>]]> The attenuated Salmonella typhimurium χ4550 strain was used to harbour a reconstructed bicistronic DNA vaccine against porcine rotavirus, which carried the rotavirus nonstructural protein 4 (NSP4) and VP7 genes simultaneously. Using a balanced lethal system, the kanamycin resistance gene of expressing eukaryotic plasmids pVAX1 and pVAXD were replaced by the aspartate β-semialdehyde dehydrogenase (asd) gene. The NSP4 cleavage product (259-525) of rotavirus OSU strain and VP7 full-length genes were amplified by reverse transcription polymerase chain reaction and then inserted into the eukaryotic single-expression plasmid, pVAX1-asd, and the eukaryotic dual-expression plasmid, pVAXD-asd, respectively. The recombinant plasmids pVAX1-asd-NSP4, pVAX1-asd-VP7 and pVAXD-asd-NSP4-VP7 were transformed into the attenuated S. typhimurium χ4550 strain by electrotransformation. An indirect immunofluorescence assay of the expressed COS-7 cell suggested that the recombinant S. typhimurium χ4550 strain was constructed successfully. The recombinant S. typhimurium χ4550 strain was orally administered to BALB/c mice. The group immunised with dual-expression plasmids produced a significantly higher level of serum Immunoglobulin G (IgG) and intestinal Immunoglobulin A (IgA) than the group immunised with single-expression plasmids. These results indicated that eukaryotic bicistronic plasmid DNA vaccines could be successfully constructed to enhance humoural, mucosal and cellular immune response against rotavirus infection. <![CDATA[<b>Temporal and spatial history of Rift Valley fever in South Africa</b>: <b>1950 to 2011</b>]]> Several outbreaks of Rift Valley fever (RVF) have been documented in South Africa since it first occurred in the country in 1950. However, there is no comprehensive account of the timing, location and extent of all known outbreaks. As part of a study investigating the epidemiology of RVF in South Africa, a full history of outbreaks was compiled using references to the disease in South Africa from scientific literature, annual reports, disease reports and animal disease databases. The geographic location and temporal occurrence of each outbreak were recorded as accurately as allowed by the available records. The result was a better and more complete picture than has hitherto been available of the spatial and temporal distribution of RVF in South Africa for the period between 1950 and 2011. Several smaller outbreaks which had not been described previously in literature were documented. Extensive outbreaks occurred in the central interior of the country (Free State, Eastern Cape and Northern Cape provinces), interspersed with smaller outbreaks or long intervening periods of absence, whilst smaller outbreaks occurred in the eastern part of the country (KwaZulu-Natal, Mpumalanga and Gauteng). <![CDATA[<b> Construction and immunogenicity of a</b> Δ<b>apxIC/ompP2 mutant of <i>Actinobacillus pleuropneumonias</i> and <i>Haemophilus parasuis</i></b>]]> The apxIC genes of the Actinobacillus pleuropneumoniae serovar 5 (SC-1), encoding the ApxI-activating proteins, was deleted by a method involving sucrose counter-selection. In this study, a mutant strain of A. pleuropneumoniae (SC-1) was constructed and named AapxIC/ ompP2. The mutant strain contained foreign DNA in the deletion site of ompP2 gene of Haemophilus parasuis. It showed no haemolytic activity and lower virulence of cytotoxicity in mice compared with the parent strain, and its safety and immunogenicity were also evaluated in mice. The LD50 data shown that the mutant strain was attenuated 30-fold, compared with the parent strain (LD50 of the mutant strain and parent strain in mice were determined to be 1.0 x 10(7) CFU and 3.5 x 10(5) CFU respectively). The mutant strain that was attenuated could secrete inactivated ApxIA RTX toxins with complete antigenicity and could be used as a candidate live vaccine strain against infections of A. pleuropneumoniae and H. parasuis. <![CDATA[<b>Occurrence of <i>Tetracampos ciliotheca</i> and <i>Proteocephalus glanduligerus</i> in <i>Clarias gariepinus</i> (Burchell, 1822) collected from the Vaal Dam, South Africa</b>]]> Cestodes are parasitic flatworms that live in the digestive tract of vertebrates as adults and often in the liver, muscle, haemocoel, mesentery and brain of various animals as larval stages. To identify the cestodes infecting Clarias gariepinus Burchell, 1822 (sharptooth catfish) in the Vaal Dam, a total of 45 host specimens were collected with the aid of gill nets between October 2011, January and April 2012. The fish were sacrificed and examined for cestode parasites. Two adult cestodes, Tetracampos ciliotheca Wedl, 1861 (prevalence 86.7%, mean intensity = 15, n = 45) and Proteocephalus glanduligerus (Janicki, 1928) (prevalence 51.1%, mean intensity = 5, n = 45) were found in the intestines of the catfish. Both T. ciliotheca and P. glanduligerus are new locality records. There were statistically insignificant differences in the infection of the male and female C. gariepinu. Fish with standard length ranging from 40 cm - 54 cm (&gt;3 years) had the highest prevalence and mean intensity while those ranging from 10 cm - 24 cm (< 1 year) had the lowest prevalence and mean intensity for both cestodes. The study highlights the importance of changing feeding habits of C. gariepinus with age on the prevalence and mean intensity of the two gastrointestinal cestode parasites. <![CDATA[<b>Comparison of pathogenic domains of rabies and African rabies-related lyssaviruses and pathogenicity observed in mice</b>]]> Several lyssavirus species occur in Africa (Rabies virus, Lagos bat virus, Mokola virus, Duvenhage virus, Shimoni bat virus and Ikoma lyssavirus), displaying a high sequence diversity between isolates belonging to the same species. There is limited information about comparative pathogenesis of these African lyssaviruses and this precludes authoritative opinion on the potential public and veterinary health impact. In this study, an analysis of representative African lyssaviruses attempted to correlate viral genomic sequence similarities and differences with the corresponding pathogenic profiles observed in mice. The study demonstrated that the virus isolates evaluated could be lethal to mice when introduced intramuscularly and that different isolates of the same lyssavirus species differ in their virulence. Using real-time polymerase chain reaction (PCR), viral RNA was detected in brain tissue, but no viral RNA was detected in the salivary glands or blood of mice that succumbed to infection. Comparison of known pathogenic domains indicated that pathogenicity is likely to be dependent on multiple domains. Cumulatively, our results re-emphasised the realisation that the pathogenicity of a lyssavirus species cannot be deduced based on studies of only a single isolate of the species or a single pathogenic domain. <![CDATA[<b>Morphological identification of parasitic nematode infective larvae of small ruminants and cattle</b>: <b>A practical lab guide</b>]]> In 2004, a new concept was introduced for simplifying identification of larvae of the common I nematodes of cattle, sheep and goats that comprises estimates of the lengths of the sheath tail extensions of infective third-stage larvae (L3) of each genus and/or species to that of I: Trichostrongylus spp., instead of having to be dependent only on measurements in micrometre. For example, if the mean length of the sheath tail extension (the extension of the sheath caudad, beyond the caudal tip of the larva) of Trichostrongylus colubriformis and Trichostrongylus axei is assumed to be 'X', then that of Haemonchus contortus is 2.0-2.7 'X' - a difference that is I not difficult to estimate. An additional new approach suggested now, particularly for L3 of species and/or genera difficult to differentiate (such as Chabertia ovina and Oesophagostomum II: columbianum), is to estimate the proportion of the larval sheath tail extension comprising a terminal thin, whip-like filament. For the experienced person, it is seldom necessary to measure I: more than one or two sheath tail extensions of L3 in a mixed culture, because the identity of most of the remaining L3 can thereafter be estimated in relation to those measured, without having to take further measurements. The aim of this article was to present the novel approach in the form of a working guide for routine use in the laboratory. To facilitate identification, figures and a separate organogram for each of small ruminants and cattle have been added to I: illustrate the distinguishing features of the common L3. <![CDATA[<b>Prevalence of mastitis in dairy cows from smallholder farms in Zimbabwe</b>]]> A cross-sectional study was conducted to determine the prevalence of sub-clinical and clinical mastitis and the associated factors in cows from selected smallholder dairy farms in Zimbabwe. Physical examinations were conducted on all lactating cows for evidence of signs of clinical mastitis. Composite milk samples were collected from all lactating cows for bacterial culture and somatic cell counting. Cows were categorised as clinical if they exhibited clinical features of mastitis, or sub-clinical if no apparent signs were present but they had a positive bacterial isolation and a somatic cell count of at least 300 x 10³ cells/mL. Farm-level factors were obtained through a structured questionnaire. The association of mastitis and animal-and herd-level factors were analysed using logistic regression. A total of 584 animals from 73 farms were tested. Overall, 21.1% (123/584) had mastitis, 16.3% (95/584) had sub-clinical mastitis and 4.8% (28/584) had clinical mastitis. Herd-level prevalence was 49.3%. Coagulase-negative staphylococci (27.6%), Escherichia coli (25.2%), Staphylococcus aureus (16.3%), Klebsiella spp. (15.5%) and Streptococcus spp. (1.6%) were the most common isolates. In individual cows, pure dairy herds (OR = 6.3) and dairy crosses (OR = 3.1) were more likely to have mastitis compared to Mashona cows. Farms that used pre-milking teat dipping were I: associated with reduced mastitis prevalence. Further research is needed on the prevalence of mastitis and a comparison of data for both smallholder and commercial dairy farms in all regions of Zimbabwe should be undertaken. <![CDATA[<b>Prevalence of brucellosis in dairy cattle from the main dairy farming regions of Eritrea</b>]]> In order to get a reliable estimate of brucellosis prevalence in Eritrean dairy cattle, a cross- | sectional study was carried out in 2009. The survey considered the sub-population of dairy cattle reared in modern small- and medium-sized farms. Samples were screened with the Rose Bengal test (RBT) and positive cases were confirmed with the complement fixation test (CFT). A total of 2.77% (417/15 049; Credibility Interval CI: 2.52% - 3.05%) of the animals tested in this study were positive for antibodies to Brucella species, with a variable and generally I: low distribution of positive animals at regional level. The highest seroprevalence was found in the Maekel region (5.15%; CI: 4.58% - 5.80%), followed by the Debub (1.99%; CI: 1.59% -2.50%) and Gash-Barka (1.71%; CI: 1.34% - 2.20%) regions. Seroprevalence at sub-regional levels was also generally low, except for two sub-regions of Debub and the sub-region Haicota I: from the Gash-Barka region. Seroprevalence was high and more uniformly distributed in the Maekel region, namely in the Asmara, Berik and Serejeka sub-regions. Considering the overall low brucellosis prevalence in the country, as identified by the present study, a brucellosis I: eradication programme for dairy farms using a test-and-slaughter policy would be possible. However, to encourage the voluntary participation of farmers to the programme and to raise their awareness of the risks related to the disease for animals and humans, an extensive public awareness campaign should be carefully considered, as well as strict and mandatory dairy movement control. <![CDATA[<b>A cost-benefit model comparing the California Milk Cell Test and Milk Electrical Resistance Test</b>]]> The indirect effects of mastitis treatment are often overlooked in cost-benefit analyses, but it | may be beneficial for the dairy industry to consider them. The cost of mastitis treatment may increase when the duration of intra-mammary infections are prolonged due to misdiagnosis of host-adapted mastitis. Laboratory diagnosis of mastitis can be costly and time consuming, therefore cow-side tests such as the California Milk Cell Test (CMCT) and Milk Electrical Resistance (MER) need to be utilised to their full potential. The aim of this study was to determine the relative benefit of using these two tests separately and in parallel. This was done using a partial-budget analysis and a cost-benefit model to estimate the benefits and costs of each respective test and the parallel combination thereof. Quarter milk samples (n = 1860) were taken from eight different dairy herds in South Africa. Milk samples were I evaluated by means of the CMCT, hand-held MER meter and cyto-microbiological laboratory analysis. After determining the most appropriate cut-off points for the two cow-side tests, the sensitivity and specificity of the CMCT (Se = 1.00, Sp = 0.66), MER (Se = 0.92, Sp = 0.62) and the tests done in parallel (Se = 1.00, Sp = 0.87) were calculated. The input data that were used for partial-budget analysis and in the cost-benefit model were based on South African figures at the time of the study, and on literature. The total estimated financial benefit of I correct diagnosis of host-adapted mastitis per cow for the CMCT, MER and the tests done in parallel was R898.73, R518.70 and R1064.67 respectively. This involved taking the expected benefit of a correct test result per cow, the expected cost of an error per cow and the cost of the test into account. The CMCT was shown to be 11% more beneficial than the MER test, whilst using the tests in parallel was shown to be the most beneficial method for evaluating the I mastitis-control programme. Therefore, it is recommended that the combined tests should be used strategically in practice to monitor udder health and promote a pro-active udder health approach when dealing with host-adapted pathogens. <![CDATA[<b>Descriptive epidemiology of African horse sickness in Zimbabwe</b>]]> A study of the prevalence of African horse sickness in horses was conducted, using records | from two private equine practices in Harare for the period 1998-2004. Results indicated a higher prevalence of the disease in horses in Zimbabwe in the late rainy season (March - May). Age of the horse was found to be a significant risk factor, with foals or yearlings appearing to be 1.80 times more likely to contract the disease compared with horses older than two years. The case fatality rate in foals or yearlings was also higher than in older age groups, but this difference was not significant. The vaccination status was an important risk factor, with vaccinated horses 0.12 times less likely to die from the disease compared with unvaccinated horses. Young, unvaccinated horses therefore seem to be the most susceptible to the disease I and have greater chances of fatality. This study highlights the importance of adequately protecting horses against African horse sickness by providing immunisation through vaccination and discusses the need to review current vaccination strategies being practiced in Zimbabwe. <![CDATA[<b>Spatial variation of epoxyscillirosidine concentrations in <i>Moraea pallida</i> (yellow tulp) in South Africa</b>]]> Moraea pallida (yellow tulp) poisoning is economically the most important intoxication of livestock in South Africa. Poisoning varies according to locality, climatic conditions and growth stage of the plant. The primary objective of this study was to determine the concentration of the toxic principle, epoxyscillirosidine, in yellow tulp leaves and to ascertain I the variability of epoxyscillirosidine concentrations within and between different locations. A secondary objective was to utilise Geographic Information Systems in an attempt to explain the variability in toxicity. Flowering yellow tulp plants were collected at 26 sampling points across 20 districts of South Africa. The leaves of five plants per sampling point were extracted and submitted for liquid chromatography/mass spectrometry analysis. A large variation in mean epoxyscillirosidine concentrations, ranging from 3.32 µg/g - 238.27 µg/g, occurred I between different geographical regions. The epoxyscillirosidine concentrations also varied tremendously between individual plants (n = 5) collected at the same sampling point, with up to a 24 times difference between the lowest and highest concentration detected. No generalised I correlation between epoxyscillirosidine concentrations and soil elemental concentrations could be established. However, samples obtained from the north-eastern part of the sampling I region tended to have higher epoxyscillirosidine concentrations compared to samples obtained from the south-western part of the sampling region. Higher toxin concentrations in the northeast were associated with statistically significant higher soil concentrations of iron, bismuth, bromide, cadmium, chromium, rubidium, tellurium, thallium, titanium and zinc, whilst soil concentrations of strontium and soil pH, were significantly lower. This study corroborated the contention that epoxyscillirosidine concentration in yellow tulp fluctuates and may explain I the variability in toxicity. <![CDATA[<b>A survey on auditing, quality assurance systems and legal frameworks in five selected slaughterhouses in Bulawayo, south-western Zimbabwe</b>]]> The purpose of this study was to explore the audits, quality assurance (QA) programmes and legal frameworks used in selected abattoirs in Zimbabwe and slaughterhouse workers' perceptions on their effectiveness. Data on slaughterhouse workers was gathered through a self-completed questionnaire and additional information was obtained from slaughterhouse and government records. External auditing was conducted mainly by the Department of Veterinary Public Health with little contribution from third parties. Internal auditing was restricted to export abattoirs. The checklist used on auditing lacked objective assessment criteria and respondents cited several faults in the current audit system. Most respondents (> 50.0%) knew the purposes and benefits of audit and QA inspections. All export abattoirs had QA programmes such as hazard analysis critical control point and ISO 9001 (a standard used to certify businesses' quality management systems) but their implementation varied from minimal to nil. The main regulatory defect observed was lack of requirements for a QA programme. Audit and quality assurance communications to the selected abattoirs revealed a variety of non-compliances with most respondents revealing that corrective actions to audit (84.3%) and quality assurance (92.3%) shortfalls were not done. A high percentage of respondents indicated that training on quality (76.8%) and regulations (69.8%) was critical. Thus, it is imperative that these abattoirs develop a food safety management system comprising of QA programmes, a microbial assessment scheme, regulatory compliance, standard operating procedures, internal and external auditing and training of workers. <![CDATA[<b>Histomorphometrical and ultrastructural study of the effects of carbendazim on the magnum of the Japanese quail <i>(Coturnix coturnix japonica)</i></b>]]> The study investigated the effect of various doses of carbendazim on the morphology of the magnum of the Japanese quail. No morphological changes were observed in the magnum in birds treated with carbendazim at doses of 25 mg/kg and 100 mg/kg bodyweight. A carbendazim dose of 400 mg/kg bodyweight was the lowest dose which caused morphological changes in the magnum. Histologically, carbendazim caused pyknosis and glandular atrophy in the magnum mucosa. Carbendazim also caused significant decreases in the height of the mucosal folds, epithelial height, glandular width and glandular luminal diameter at 400 mg/kg and 800 mg/kg (p < 0.05). At ultrastructural level, dose-dependent deciliation was observed. Pyknotic nuclei, dilated cisternae of rough endoplasmic reticulum, swollen mitochondria, numerous vacuoles and lysosomes in the luminal and glandular epithelia were identified. The observed degenerative changes could be due to cytoskeletal disruption caused by carbendazim toxicity. Degeneration of the luminal and glandular cells in the magnum pose a potential threat to the egg production and reproduction of exposed birds. <![CDATA[<b>Development of real-time reverse transcription polymerase chain reaction assays to quantify insulin-like growth factor receptor and insulin receptor expression in equine tissue</b>]]> The insulin-like growth factor system (insulin-like growth factor 1, insulin-like growth factor 2, insulin-like growth factor 1 receptor, insulin-like growth factor 2 receptor and six insulinlike growth factor-binding proteins) and insulin are essential to muscle metabolism and most aspects of male and female reproduction. Insulin-like growth factor and insulin play important roles in the regulation of cell growth, differentiation and the maintenance of cell differentiation in mammals. In order to better understand the local factors that regulate equine physiology, such as muscle metabolism and reproduction (e.g., germ cell development and fertilisation), real-time reverse transcription polymerase chain reaction assays for quantification of equine insulin-like growth factor 1 receptor and insulin receptor messenger ribonucleic acid were developed. The assays were sensitive: 192 copies/µL and 891 copies/µL for insulin-like growth factor 1 receptor, messenger ribonucleic acid and insulin receptor respectively (95% limit of detection), and efficient: 1.01 for the insulin-like growth factor 1 receptor assay and 0.95 for the insulin receptor assay. The assays had a broad linear range of detection (seven logs for insulinlike growth factor 1 receptor and six logs for insulin receptor). This allowed for analysis of very small amounts of messenger ribonucleic acid. Low concentrations of both insulin-like growth factor 1 receptor and insulin receptor messenger ribonucleic acid were detected in endometrium, lung and spleen samples, whilst high concentrations were detected in heart, muscle and kidney samples, this was most likely due to the high level of glucose metabolism and glucose utilisation by these tissues. The assays developed for insulin-like growth factor 1 receptor and insulin receptor messenger ribonucleic acid expression have been shown to work on equine tissue and will contribute to the understanding of insulin and insulin-like growth factor 1 receptor physiology in the horse. <![CDATA[<b>Prevalence of peste des petits ruminants in the arid zone in the Republic of Niger</b>]]> The study aimed to determine the prevalence of peste des petits ruminants in the arid zone (Niamey, Tillabéry and Tahoua) of the Republic of Niger. A serological survey was conducted and 519 serum samples were collected from 253 unvaccinated sheep and 266 unvaccinated goats. The sample included 340 female animals (168 sheep and 172 goats) and 160 kids and lambs (78 lambs and 82 kids). A competitive enzyme-linked immunosorbent assay yielded an overall seroprevalence of 45.0%. The prevalence in sheep was 42.0% compared with 47.9% in goats. The seroprevalence observed amongst small ruminants in Tahoua (49.8%) and Tillabéry (46.6%) was significantly higher (p = 0.001) than that observed in animals from Niamey (25.1%). It was also higher (p = 0.04) in sheep younger than two years (51.8%) than in adults (37.6%). Conversely, the seroprevalence showed no significant difference between male animals (35.8% in sheep; 50.1% in goats) and female animals (45.1% in sheep; 46.4% in goats). The prevalence of the disease observed amongst the sheep and goat populations confirms the continued danger of this disease in the areas studied. It is therefore necessary to develop strategies such as improving livestock services, providing effective vaccines and implementing a vaccination programme for an effective control of the disease in sub-Saharan Africa. <![CDATA[<b>Molecular surveillance of <i>Theileria ovis, Theileria lestoquardi</i> and <i>Theileria annulata</i> infection in sheep and ixodid ticks in Iran</b>]]> A molecular study was undertaken to detect Theileria ovis, Theileria lestoquardi and Theileria annulata in sheep and tick vectors. Investigation was conducted from 2010 to 2011 in the south of Khorasan Razavi Province, Iran. A total of 150 blood samples were collected from 30 different sheep flocks. In addition, ixodid ticks were sampled from the same flocks. The stained blood smears were microscopically examined for the presence of piroplasms and a semi-nested polymerase chain reaction-restriction (PCR) was used for subsequent molecular speciation. Salivary glands were isolated from the ticks and subsequently analysed by semi-nested PCR. polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to differentiate between T. lestoquardi and T. annulata from PCR-positive samples. Theileria species infection was microscopically detected in 18.6% of blood smears. The presence of T. ovis and T. lestoquardi or T. annulata was detected by semi-nested PCR in 58.6% and 6.6% of blood samples respectively. In total, 169 ixodid ticks were collected from different areas of the province. The most prevalent ticks were Rhipicephalus turanicus (n = 155; 91.7% of the total), followed by Hyalomma anatolicum anatolicum (n = 8; 4.7%) and Hyalomma marginatum turanicum (n = 6; 3.5%). From an organ pooling of 33 ticks, three pools of salivary glands from R. turanicus were positive for Theileria species by semi-nested PCR. Of the three R. turanicus samples testing positive for Theileria species, two (6.1%) were positive for T. ovis and one (3.0%) for T. lestoquardi or T.annulata. Amongst the 11 PCR-positive samples for T. lestoquardi or T. annulata, 10 were positive for T. lestoquardi and one sample was positive for both T. lestoquardi and T. annulata using PCR-RFLP. The results also demonstrated that PCR-RFLP could be used for the detection of T. ovis. Based on the results, it can be concluded that T. ovis has a higher prevalence than T. lestoquardi, and that R. turanicus could be a possible vector for T. ovis and T. lestoquardi. Finally, the PCR-RFLP based on Msp1 restriction enzyme is a simple method for differentiation of Theileria species in sheep and ixodid ticks. <![CDATA[<b>The incursion, persistence and spread of peste des petits ruminants in Tanzania: Epidemiological patterns and predictions</b>]]> Peste des petits ruminants virus, which causes a severe disease in sheep and goats, has only recently been officially declared to be present in Tanzania. An epidemiological study was carried out between September 2008 and October 2010 to investigate the incursion, persistence and spread of the virus in Tanzania. The investigation involved serosurveillance, outbreak investigation and computation of epidemiological indices such as the effective reproductive number, persistence and the threshold level for vaccination. Field and molecular epidemiological techniques were applied to isolate, characterise and trace the origin of the virus in Tanzania. A total of 2182 serum samples from goats and 1296 from sheep from 79 villages across 12 districts were investigated. Village-level prevalence of infection was variable (0.00% - 88.00%) and was higher in pastoral than in agro-pastoral villages. The overall antibody response to the virus was 22.10% (CI95% = 20.72% - 23.48%). About 68.00% and 73.00% of seropositive goats and sheep, respectively, did not show clinical signs. The proportion of seropositive animals differed significantly (p < 0.001) between age groups, sex and farming practices. Real-time polymerase chain reaction results showed that the isolated strains belong to lineage III, whose origin is in East Africa and the Middle East. This indicates that one of the northern neighbouring countries is most likely the source of infection. The computed overall effective reproductive number, the threshold level of vaccination necessary to eradicate the disease and persistence were 4.75% and 98.00%, respectively. These estimates indicate that achieving elimination of the peste des petits ruminants virus from pastoral flocks will require significant effort and development of highly effective intervention tools. <![CDATA[<b>Immunohistochemical studies of the enteric nervous system and interstitial cells of Cajal in the canine stomach</b>]]> The distribution of interstitial cells of Cajal (ICC), the probable pacemakers in gastrointestinal motility, was investigated using an antigenic marker of gastric ICC known as C-Kit. Antiserum raised against the general neuronal marker protein gene peptide 9.5 (PGP) as well as the nitrergic neuronal marker neuronal nitric oxide synthase (nNOS) were used to investigate the distribution of gastric nerves. Polyclonal goat anti-human C-Kit was reliable in labelling ICC in the stomach. Two classes of ICC were identified according to their distribution: ICC-MY distributed around the periphery of myenteric ganglia and ICC-IM in the circular and longitudinal muscle layers. The neuronal marker PGP was reliably consistent in revealing the density and distribution of the enteric nervous system. Density of nerve fibres was higher in circular smooth muscle than in longitudinal smooth muscle. From nNOS immunohistochemistry, it is evident that inhibitory (nitrergic) nerves constitute a substantial fraction of the enteric nervous system. <![CDATA[<b>Notes on the occurrence of <i>Trypanosoma</i> sp. (Kinetoplastida</b>: <b>Trypanosomatidae) in freshwater fishes from South Africa</b>]]> A total of 257 fishes from four families, Clariidae, Cichlidae, Cyprinidae and Schilbeidae were I collected from three localities: the Sand River Dam, Swaziland; the Nylsvlei Nature Reserve, South Africa and the Vaal Dam and Vaal River Barrage, South Africa. Only fishes (n = 154) from Clariidae and Cichlidae were found to be infected with trypanosomes. A total of 221 Clarias gariepinus (Burchell 1822) were collected from the Vaal Dam and Vaal Barrage area, South Africa. Of these, 74% (89/121) were infected with trypanosomes from the Vaal Dam and 63% (63/100) from the Vaal River Barrage, with no seasonal infection pattern. A prevalence of 25% (1/4) was found in C. gariepinus from the Sand River Dam, Swaziland, and a 50% (1/2) prevalence was found in Tilapia sparrmanii from the Nylsvlei Nature Reserve, South I: Africa. Standard measurements conformed closely to the morphometric and morphological descriptions of Trypanosoma mukasai. This article provides new locality records for T. mukasai II: from the Vaal Dam, Vaal River Barrage and Nylsvlei Nature Reserve (South Africa) and the Sand River Dam (Swaziland). Tilapia sparrmanii collected in the Sand River Dam in Swaziland is also noted as a new host record. <![CDATA[<b>Nematodes of the small intestine of African buffaloes, <i>Syncerus caffer,</i> in the Kruger National Park, South Africa</b>]]> The abundance and distribution of parasitic helminths in populations of African buffaloes, | Syncerus caffer, have not been well documented. A total of 28 buffaloes of different ages and sexes were sampled in the Kruger National Park, South Africa, for nematodes of the small intestine. Three nematode species were identified, namely Cooperia fuelleborni, Cooperia hungi and Trichostrongylus deflexus, with C. hungi being a new country record for African buffalo in South Africa. The overall prevalence was 71% and the average number of worms was 2346 (range: 0-15 980). This is a small burden for such a large mammal. Sex, age and body condition of the buffaloes had no significant effect on worm occurrence. <![CDATA[<b>Selected haematological changes in <i>Clarias gariepinus</i> (Burchell, 1822) infected with a <i>Trypansosoma</i> sp. from the Vaal Dam, South Africa</b>]]> The use of haematological techniques to assess fish health is generally accepted. The aim of the current study was to determine selected haematological changes that occur in Clarias gariepinus (Burchell, 1822). infected with trypanosomes. Blood films were prepared according to standard techniques to confirm trypanosome infections and whole blood was collected, the serum and plasma separated, and prepared for albumin and total protein concentration analysis. Plasma albumin levels were significantly higher in infected wild caught fish than in uninfected wild caught fish and uninfected breeding stock. Serum albumin levels were significantly lower in infected wild caught fish when compared to uninfected breeding stock. The total plasma and serum protein levels were within the normal range for C. gariepinus, that is, 3 g - 6 g/100 mL. The total plasma protein levels varied significantly between the three groups. However, the total serum protein levels were only significantly different between uninfected breeding stock and uninfected wild caught fish, as well as uninfected breeding stock and infected wild caught fish. The total protein levels were significantly higher in infected wild caught fish than in the other groups, a possible indication of an infection or inflammatory host response. <![CDATA[<b>Concomitant fungal and <i>Mycobacterium bovis</i> infections in beef cattle in Kenya</b>]]> Bovine tuberculosis is an important zoonosis and accurate diagnosis is important for its surveillance. Post-mortem diagnosis may, however, be compromised by lesions caused by other pathogens. In an investigation on its prevalence in slaughter cattle in Kenya, Mycobacterium bovis and dimorphic fungi were inadvertently identified separately or concurrently in tuberculous lesions. Beef carcasses were inspected for lesions in two abattoirs in Nairobi. Tissues with lesions were collected and transported to the laboratory. Smears of lesions were stained by acid-fast procedure and examined microscopically. Lesions were cultured in Löwenstein-Jensen (LJ) and in BBL™ Mycobacterium growth indicator tubes (MGIT) media. Mycobacteria isolates in LJ medium were identified by DNA typing. Smears of BBL TM MGIT cultures were acid-fast stained and examined microscopically. Tissue sections were stained with periodic acid-Schiff reagent before examination. Of the 929 carcasses examined, 176 had granulomatous lesions. Dimorphic fungi were detected as acid-fast negative cells in 58 (32.9; 33.5%) of the lesion smears, either alone (29.0; 16.4%) or concurrently with acid-fast bacilli (29.0; 16.4%). The fungi were also detected in some BBL TM MGIT-culture smears and lesioned tissue sections. The fungi were identified, by means of cellular morphology, as Paracoccidioides brasiliensis and Blastomyces dermatitidis. A total of 64 isolates of mycobacteria were recovered in LJ medium, 19 of which were identified as M. bovis. The present report documents native P. brasiliensis infections outside the presumed endemic region and B. dermatitidis infections in a livestock animal. The findings further indicate the importance of dimorphic fungi as a differential diagnosis of bovine tuberculosis in the region. <![CDATA[<b>Biliary and plasma copper and zinc in pregnant Simmental and Angus cattle</b>]]> Three each of 3-year-old Angus and Simmental heifers, surgically modified to collect bile, were used to measure the effects of pregnancy and breed on bile flow, biliary copper and zinc excretion and plasma copper and zinc concentrations. Bile copper excretion was significantly higher at 7-mo of pregnancy when samples from both breeds were pooled. From then onwards it declined to its lowest, one week post-partum. During pregnancy, plasma copper concentration increased slightly, reaching its highest level at 7-mo of pregnancy and then decreased slightly until full term. In pooled samples from both breeds, the correlation between increase in bile copper excretion and plasma copper concentration from 0 to 7-mo of pregnancy was high (r = 0.85) and significant (p < 0.05). Plasma zinc concentration decreased to the lowest level around 6-mo of pregnancy but increased thereafter until full term. In cows that were dried off one week after parturition, major shifts in bile and plasma copper and zinc parameters occurred at one week following and these coincided with a marked decline of bile flow and bile copper and zinc excretion. By 3-mo post-partum, biliary copper and zinc excretion and plasma copper and zinc concentrations had reached levels observed prior to pregnancy. When the data from all samples were pooled, the bile flow and bile copper excretion were significantly (p < 0.05) higher in Simmental, and plasma copper and zinc concentration higher in the Angus. <![CDATA[<b>Assessment of acquired immune response to <i>Rhipicephalus appendiculatus</i> tick infestation in different goat breeds</b>]]> Changes in serum gamma globulin levels, numbers of replete female ticks and engorged tick mass were used as parameters to monitor the acquired immune response (antibody mediated immune response) elicited by Rhipicephalus appendiculatus adult tick infestations. Three consecutive Rhipicephalus appendiculatus adult tick infestations were applied to South African Indigenous goats (Nguni), Saanen goats and cross-bred goats (Saanen goats crossed with South African Indigenous goats [Nguni]) under laboratory conditions. During the three consecutive Rhipicephalus appendiculatus adult tick infestations the serum gamma globulin levels increased in all three breeds, whilst the mean replete female tick numbers and engorged tick mass decreased. Even though all three goat breeds exhibited an acquired immune response, the South African Indigenous goats (Nguni) response was significantly higher than that of the Saanen and cross-bred goats. However, the acquired immune response elicited by Saanen goats was significantly lower when compared with cross-bred goats. <![CDATA[<b>A serological survey of brucellosis in wild ungulate species from five game parks in Zimbabwe</b>]]> A retrospective serosurvey was carried out between 2009 and 2012 to detect antibodies to Brucella spp. in free-ranging African wildlife ungulates from five selected game parks in Zimbabwe. Samples were drawn from wildlife-livestock interface and non-interface areas in Zimbabwe. A total of 270 serum samples from four different species, namely African buffalo (Syncerus caffer) (n =106), impala (Aepyceros melampus) (n = 72), black rhinoceros (Diceros bicornis) (n = 45) and white rhinoceros (Ceratotherium simum) (n = 47), were tested. The percentage of positive samples was 17.0% in buffalo (18/106; 95% CI: 9.72% - 24.1%) and 1.4% in impala (1/72; 95% CI: 0% - 4.2%). No antibodies to Brucella spp. were detected in the two rhinoceros species. The difference in the percentage of seropositive cases between buffalo and impala was significant (p < 0.05). Seropositivity to Brucella spp. was higher (19.1%) in adult buffalo compared with juveniles and sub-adults younger than six years (5.9%). Further, seropositivity was marginally higher (20.4%) in animals from wildlife-livestock interface areas than in those from non-interface areas (13.45%; OR = 1.45) although the difference was not statistically significant. The study showed that brucellosis could be more widespread in buffalo and may circulate in this species independently in the absence of contact with cattle, whilst rhinoceros may be considered less susceptible to brucellosis. The role of the wildlife-livestock interface in the epidemiology of brucellosis in wildlife and livestock is probably overstated but needs to be explored further. <![CDATA[<b>Descriptions of diplostomid metacercariae (Digenea: Diplostomidae) from freshwater fishes in the Tshwane area</b>]]> The metacercarial (larval) stages of diplostomid digeneans are known to inhabit freshwater fish, causing tissue damage in the process. Due to their widespread diversity, little is known about their life cycle. The classification of these parasitic stages to the species level using only the morphology is very challenging due to the lack of genitalia; they are regarded to be the most important structures in the identification of these organisms. In this study, additional morphological information through light and scanning electron microscopy is given for two different diplostomids found in the cranial cavity of Clarias gariepinus and the vitreous chambers of Tilapia sparrmanii and Pseudocrenilabrus philander. The diplostomid metacercaria inhabiting the cranial cavity of Clarias gariepinus was morphologically identified as Diplostomulum (Tylodelphys) mashonense and an unknown metacercaria of the genus Diplostomum was found in the vitreous chambers of Pseudocrenilabrus philander and Tilapia sparrmanii. Both parasitic species' 28S recombinant deoxyribonucleic acid genomic regions were successfully amplified using Dig 125/1500R primer pairs. The assay yielded a product of approximately 1300 base pairs as seen on the gel images. There were 14 nucleotide differences over the entire analysed sequences resulting in a 1.1% (14/1273) nucleotide difference. In line with the morphological characteristics of these parasites, there seemed to be a slight difference in their genetic makeup. The application of molecular techniques on digenetic trematodes seems very promising and may yield great potential in future descriptions of morphologically similar parasitic species. <![CDATA[<b>A review of the epidemiology and control of gastrointestinal nematode infections in cattle in Zimbabwe</b>]]> In this review, the main gastrointestinal nematodes infecting cattle in Zimbabwe and the epidemiological factors influencing their occurrence are reviewed and discussed. Nineteen gastrointestinal nematode species that belong to seven families have been found to occur in cattle in Zimbabwe. The main genera reported to date are Cooperia, Haemonchus, Trichostrongylus and Oesophagostomum and the dominant species are Cooperia pectinata, Cooperia punctata, Haemonchus placei and Trichostrongylus axei. The mixed infection by several species from the genera is the cause of parasitic gastroenteritis in cattle in Zimbabwe. Production and husbandry practices, season, host age and environment are considered to be the main factors that influence gastrointestinal nematode infection in cattle. The geographical distribution of the gastrointestinal nematodes is also reviewed in relation to the climatic conditions of the country. Various control options are discussed and how they are applicable to the Zimbabwean situation. Based on reports and existing data on the epidemiological features of the gastrointestinal nematode infection in cattle, practical control measures are critically reviewed and recommendations are made for a national control programme. <![CDATA[<b>Lack of evidence for safe vaccination with the Muguga cocktail in Sudan</b>]]> In this review, the main gastrointestinal nematodes infecting cattle in Zimbabwe and the epidemiological factors influencing their occurrence are reviewed and discussed. Nineteen gastrointestinal nematode species that belong to seven families have been found to occur in cattle in Zimbabwe. The main genera reported to date are Cooperia, Haemonchus, Trichostrongylus and Oesophagostomum and the dominant species are Cooperia pectinata, Cooperia punctata, Haemonchus placei and Trichostrongylus axei. The mixed infection by several species from the genera is the cause of parasitic gastroenteritis in cattle in Zimbabwe. Production and husbandry practices, season, host age and environment are considered to be the main factors that influence gastrointestinal nematode infection in cattle. The geographical distribution of the gastrointestinal nematodes is also reviewed in relation to the climatic conditions of the country. Various control options are discussed and how they are applicable to the Zimbabwean situation. Based on reports and existing data on the epidemiological features of the gastrointestinal nematode infection in cattle, practical control measures are critically reviewed and recommendations are made for a national control programme.