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Canine leishmaniasis is a vector-borne disease caused by protozoa of the genus Leishmania that affect dogs, humans and wildlife. Sandflies of the genera Phlebotomus and Lutzomyia are the primary vectors. Canine leishmaniasis is an exotic and controlled disease in South Africa. The main purpose of our risk assessment study was to evaluate the likelihood that this exotic disease could enter and be established in South Africa through importation of live dogs. Risk analysis to the spread of the disease follows the World Organization for Animal Health (OIE) formal method of quantitative risk assessment documented as a step-by-step process. We have identified and discussed 11 possible risk factors involved in three steps for final assessment. The annual average number of diagnostic tests performed on imported dogs from 44 countries for 2011-2015 was 1158. Leishmania is reported to occur in 21/44 (47.7%) exporting countries. A total of 71.1% of Leishmania positive dogs were imported from these endemic countries. The yearly percentage of Leishmania positive dogs ranged from 0.2% to 2%. Three confirmed clinical and fatal cases of leishmaniasis in dogs of unidentified origin have been reported by our laboratory and the state veterinarians. The disease has been reported in neighbouring countries as well as the putative sandfly vectors. This study concluded that the risk for the introduction and degree of uncertainty of Leishmania in imported dogs in South Africa are moderate. Risk mitigation and recommendations such as investigations into possible occurrence of autochthonous leishmaniasis in the country, surveillance in its wildlife reservoirs and systematic surveillance of sandfly populations are discussed.

Quantitative real-time PCR assays previously developed for the detection of Theileria equi and Babesia caballi, were combined in a single multiplex TaqMan qPCR platform for the simultaneous detection of both heamoprotozoan parasites in equids. The multiplex equine piroplasmosis (M-EP) qPCR assay was shown to be efficient and specific. The detection limit was determined to be 1.4 × 10 -4 % parasitized erythrocytes (PE) for T. equi and 2.8 × 10 -4 % PE for B. caballi. The effect of differential DNA concentrations on the outcome of the M-EP qPCR for each target species was also investigated. The data demonstrated that the assay could reliably detect both targets, over a range of at least 1000-fold difference in target concentrations, without loss of sensitivity. The assay was subsequently evaluated on 243 field samples collected from areas where limited tick control strategies were implemented. The IFAT detected circulating T. equi and B. caballi antibodies in 100% and 92% of the samples, respectively. The M-EP qPCR assay detected T. equi parasite DNA in 98% of the samples, while B. caballi could only be detected in 6% of the samples tested, confirming that B. caballi infections generally occur at extremely low parasitaemias that rarely exceed 1%. The developed M-EP qPCR assay therefore serves as a reliable tool for the rapid diagnosis and epidemiological survey of equine piroplasmosis., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A total of 719 serum samples collected from clinically healthy cattle from eight provinces located in different districts of South Africa were examined by the indirect enzyme-linked immunosorbent assay (ELISA) and the standard indirect fluorescent antibody test (IFAT) to determine the serological prevalence of Babesia bovis and Babesia bigemina. The results showed that 35.3% and 39.7% of cattle were positive for B. bovis and 30% and 36.5% were positive for B. bigemina antibodies on ELISA and IFAT, respectively. Mixed infections were detected in 18.2% and 26.3% of the samples using ELISA and IFAT, respectively. Consequently, the ELISAs with recombinant B. bovis spherical body protein-4 (BbSBP-4) and B. bigemina C-terminal rhoptry-associated protein-1 (BbigRAP-1/CT) were proven to be highly reliable in the serological diagnoses of bovine babesiosis in South African cattle, as evidenced by the significant concordance rates when the results were compared to those of IFAT. Moreover, the serological prevalence was significantly different among the tested provinces, in which the ranges exhibited between 15% and 73% for B. bovis infection and between 13% and 54% for B. bigemina infection. High sero-positive rates were present in Mpumalanga and KwaZulu-Natal provinces, while the lowest rate was in the North West province. Our data provide important information regarding the current seroprevalence of bovine babesiosis in South Africa, which might be beneficial in developing rational strategies for disease control and management., (Copyright © 2011 Elsevier B.V. All rights reserved.)

A competitive enzyme-linked immunosorbent assay (cELISA) based on a broadly conserved, species-specific, B-cell epitope within the C terminus of Babesia bigemina rhoptry-associated protein 1a was validated for international use. Receiver operating characteristic analysis revealed 16% inhibition as the threshold for a negative result, with an associated specificity of 98.3% and sensitivity of 94.7%. Increasing the threshold to 21% increased the specificity to 100% but modestly decreased the sensitivity to 87.2%. By using 21% inhibition, the positive predictive values ranged from 90.7% (10% prevalence) to 100% (95% prevalence) and the negative predictive values ranged from 97.0% (10% prevalence) to 48.2% (95% prevalence). The assay was able to detect serum antibody as early as 7 days after intravenous inoculation. The cELISA was distributed to five different laboratories along with a reference set of 100 defined bovine serum samples, including known positive, known negative, and field samples. The pairwise concordance among the five laboratories ranged from 100% to 97%, and all kappa values were above 0.8, indicating a high degree of reliability. Overall, the cELISA appears to have the attributes necessary for international application.

The possible role of Rhipicephalus appendiculatus and Amblyomma hebraeum in the mechanical and transstadial transmission of bovine leukosis virus (BLV) was investigated. BLV-free laboratory strains of R. appendiculatus and A. hebraeum nymphal ticks (n = 400) were fed on a BLV-infected and a negative control bovine. At various intervals after engorgement the ticks were homogenised and injected subcutaneously into BLV-negative sheep. Adult R. appendiculatus and A. hebraeum, which had fed as nymphs on the BLV-infected bovine, were then allowed to feed on BLV-negative sheep. A control sheep was also injected intravenously with blood from the infected bovine. Only the control sheep that received blood from the BLV-positive bovine seroconverted 9 months later. All the other surviving sheep remained serologically negative during the 13 months observation period. It is suggested that the nymphal stages of these ticks probably do not play a role in the transstadial transmission of BLV in southern Africa. The significance of these results is discussed.

Natural and experimental cases of sweating sickness were treated using a hyperimmune serum as specific treatment and hyperimmune serum combined with symptomatic and supportive treatment based on the clinicopathological changes observed in cases of sweating sickness. The treatment regimens were found to be highly effective in pigs and sheep as well as in calves, although recovery in the latter species was slower.

An ELISA was developed using an SDS extract of Brucella abortus as antigen to detect antibodies in cattle sera. The antigen was stable at 4 degrees C for at least 4 years and although it gave optimal results at a 1/4,000 dilution it could detect reactors at a 1/32,000 dilution. Based on comparative CF tests on 430 sera from negative herds and 187 sera from positive herds a reading of 0,07 or less using a 492 nm filter could be considered a negative reaction. The ELISA could be a useful test to supplement the CF test but further evaluation is still required.

Studies to evaluate cardiac and pulmonary function were undertaken in 4 calves suffering from experimentally-induced heartwater. There was a marked variation in the course of the disease. Three of the calves recovered spontaneously after developing clinical signs. These included a rectal temperature in excess of 40 degrees C, anorexia and listlessness but no neurological signs. The remaining calf died 2 days after developing a fever and neurological signs. In the 3 calves that recovered, a mild hypoxemia developed during the acute stage of the disease. Arterial CO2 tension remained within normal limits, but there was a tendency towards an alkalosis. Increases in pulmonary dead space and fluctuations in venous admixture were observed. The calf that died showed similar mild changes in blood gas parameters, despite the presence of a marked reduction in minute volume, and a lung oedema was demonstrated on post-mortem examination. No marked changes in systolic and diastolic blood pressures and in right cardiac intraventricular pressures were observed. Terminally, however, there were marked decreases in stroke volume and cardiac output. These changes were associated with a sharp increase in heart rate. No primary cardiac pathology was observed on clinical and post-mortem examinations.

Experimentally-induced cases of sweating sickness in calves were used in an effort to correlate the blood chemistry with some of the known pathological changes. Results showed that the "sweating" associated with necrotic dermatitis did not alter blood electrolyte levels. Laboratory evidence of a disseminated intravascular coagulopathy was found which correlated with the microthrombi described in cases of sweating sickness. A high blood cortisol level was found in one of the animals that died from the disease and could possibly be used as a prognostic indicator in clinical cases. Recommendations are made with regard to the supportive treatment based on the clinical pathological findings.

This paper reviews the available literature on the clinical pathology and pathophysiology of heartwater and makes comparisons with unpublished results obtained from a recent study in experimentally-induced heartwater in calves. The pathophysiological changes seem to center on an increased capillary permeability the result of which is reflected most noticeably in cardiac and lung function. There is a marked drop in cardiac output in severe cases and some workers have recorded a severe drop in diastolic blood pressure in the advanced stage of the disease. Changes in lung function are variable, depending on the stage of the disease, and may change from a respiratory alkalosis in the early febrile stage to a respiratory acidosis in more advanced cases. The basic cause for the increased capillary permeability is not known. The main clinical pathological changes measured include a progressive anaemia, fluctuations in total and differential white cell count, of which an eosinopenia and a lymphocytosis are the most marked, increases in total bilirubin which coincide with darkening of plasma colour, and a drop in total serum proteins mostly shown in the albumin levels.