Darshana Morar-Leather's research while affiliated with University of Pretoria and other places

Interface areas shared by humans, domestic and wild animals may serve as high transmission contexts for Toxoplasma gondii. However, knowledge about the epidemiology of T. gondii in such areas is currently limited. The present study assessed the seroprevalence of T. gondii in different hosts from Mpumalanga, South Africa. Furthermore, we investigated the local knowledge and related practices about T. gondii by conducting a questionnaire study in the community. Blood samples were obtained and analysed for T. gondii antibodies using a commercial multispecies latex agglutination kit. The seroprevalence detected in humans (n = 160; patients showing signs of acute febrile illness), cats (n = 9), chickens (n = 336) and goats (n = 358) was 8.8%, 0.0%, 4.2% and 11.2%, respectively. Seroprevalence in impalas (n = 97), kudus (n = 55), wild dogs (n = 54), wildebeests (n = 43), warthogs (n = 97) and zebras (n = 68) was calculated at 5.2%, 7.3%, 100.0%, 20.9%, 13.4% and 9.1%, respectively. The questionnaire revealed that 63.0% of household owners were subsistence farmers, and 35.9% were pet owners. A high level of female participation was found (75.3%) when compared to male participation (24.7%). The results show a low circulation of T. gondii in the domestic cycle and suggest the presence of possible bridges between the wildlife cycle and the surrounding domestic cycle.Contribution: The study contributes to identifying transmission patterns and risk factors of T. gondii within human and animal populations. This topic fits within the scope of the journal presenting original research in veterinary science, with the focus on wild and domestic populations on the African continent on a topic of universal importance.

Amblyomma spp. ticks, known for their bright ornate appearance and aggressive hunting behaviour, are vectors of a number of important pathogens. In southern Africa, 17 Amblyomma spp. are currently documented. Of these species, Amblyomma hebraeum and Amblyomma variegatum have been well studied due to their wide geographical range and their status as competent vectors of pathogens that are of veterinary and medical importance. Studies on other Amblyomma spp. in southern Africa have been neglected, fostering ongoing debates on the validity of certain species such as Amblyomma pomposum . This study investigated the inter- and intraspecies variation of Amblyomma ticks collected in southern Africa, focusing on resolving the dispute about A. pomposum and A . variegatum as distinct species. Four tick species were collected from Angola, Mozambique, South Africa, Zambia, and Zimbabwe and were identified morphologically as Amblyomma eburneum , A . hebraeum , A . pomposum and A . variegatum using identification keys. Gene amplification was done targeting the 12S and 16S rRNA, cytochrome oxidase I, cytochrome B and internal transcribed spacer-2 genes, and Bayesian inference analyses were performed in MrBayes. These revealed little geographic structuring amongst ticks of the same species from different countries, although intraspecific variation within A. variegatum was high for the 16S and cytB loci. Our study concluds that there is insufficient molecular evidence to differentiate A . pomposum and A . variegatum from each other. We highlight the need for whole mitochondrial genome sequencing of these two species to resolve the ongoing debates. Furthermore, we propose mating and hybrid viability studies between the two species to confirm their reproductive isolation.

Heartwater, one of the major tick-borne diseases of some domestic and wild ruminants in Africa, is caused by Ehrlichia ruminantium. The genetic diversity of E. ruminantium isolates renders the available vaccine ineffective against certain virulent isolates. To better understand the E. ruminantium genotypes in South Africa, a total of 1004 Amblyomma hebraeum tick deoxyribonucleic acid (DNA) samples from cattle in three South African provinces were tested by pCS20 Sol1 real-time polymerase chain reaction (qPCR) and characterised by multilocus sequence typing (MLST) using five housekeeping genes. Out of 1004 samples tested, 222 (22%) were positive for E. ruminantium. The occurrence of E. ruminantium in Mpumalanga, KwaZulu-Natal and Limpopo provinces was 19%, 22% and 27%, respectively. The E. ruminantium positive samples were screened for housekeeping genes and sequenced. Phylogenetic analysis revealed three main lineages: clade 1 made up of worldwide isolates (eastern, southern Africa, and Caribbean isolates), clade 2 comprised only West African isolates and clade 3 consisted of Omatjenne, Kümm2 and Riverside. Some study sample sequences were not identical to any of the reference isolates. However, they could all be grouped into the worldwide clade. Genetic variation in the sequenced regions was observed in the form of single nucleotide polymorphisms (SNPs). Using MLST to characterise E. ruminantium field isolates allowed the South African genotypes to be clearly distinguished from the distinct West African isolates.Contribution: Characterisation of E. ruminantium field isolates is important for the control of heartwater and contributes to preliminary knowledge required for the development of a more practical vaccine against heartwater.

The Amblyomma genus is represented on the African continent by 24 species, out of which 17 are known to occur in different ecological niches of southern Africa. Amblyomma, known for their aggressive hunting behaviour and aptitude as pathogen vectors, are of main concern to travellers, mainly in rural and conservation areas of Africa. In this study, we highlight the overlapping distribution of Amblyomma eburneum and Amblyomma variegatum found on African buffaloes (Syncerus caffer) at Coutada 11, Central Mozambique. In total, 1,039 Amblyomma ticks were collected and morphologically identified using taxonomic keys, and genomic DNA was extracted. They were subjected to reverse line blotting for pathogen identification followed by molecular analysis (COI sequencing) of both tick species. Pathogens such as Ehrlichia ruminantium, Anaplasma centrale, Theileria sp., Babesia sp. and Rickettsia africae were detected, of which R. africae is zoonotic. Ehrlichia ruminantium, R. africae, Theileria mutans and Theileria velifera are well-established pathogens transmitted by Amblyomma ticks; however, Anaplasma spp. and Babesia spp. are not, suggesting residual parasite DNA in the bloodmeal. Little is mentioned in the literature about A. eburneum, including its role as a vector and reservoir for pathogens. In Mozambique A. eburneum is currently restricted to wildlife but the spread of the tick may be observed given the climate change that is occurring. The infection rates for the pathogens in both Amblyomma tick species were lower than expected, but this may be due to the low host density in the forest niche and the innate immunity of these hosts. With the propensity of ticks of the Amblyomma genus to form parapatric distributions, the mechanisms that allows for the overlapping distribution of these two Amblyomma species while maintaining tick species identity is of great interest.

Toxoplasma gondii (T. gondii) is a protozoan parasite, which infects a wide variety of mammals and bird species globally. In large parts of the world, this parasite is relatively well documented in wildlife species, however, this topic is poorly documented in Africa. The current review systematically explores the presence and distribution of T. gondii in African wildlife species through a key word search in PubMed, Web of Science and CAB Direct. A total of 66 records were identified and included in the qualitative analysis, of which 19 records were retained for the quantitative synthesis. The presence of T. gondii was reported in a wide range of wildlife species, found in twelve countries, spread over the African continent. The retained records report a prevalence range of 6–100% in herbivores, 8–100% in omnivores and 14–100% in carnivores. In wild felines (cheetahs, leopards, and lions) a prevalence range of 33–100% was found. Reports from South Africa, and on the presence of T. gondii in lion were most common. Overall, the results indicate the scarcity of information on T. gondii in Africa and its circulation in wildlife. The lack of knowledge on the parasite in Africa, especially in areas at the human-livestock-wildlife interface, prevents us from understanding how prevalent it is on the continent, what strains are circulating in wildlife and what the most common routes of transmission are in the different habitats in Africa.

Rickettsia africae is a gram-negative bacterium, which causes African tick bite fever (ATBF) in humans. ATBF is a febrile disease mainly affecting travellers to southern Africa. This bacterium is known to be transmitted by Amblyomma hebraeum and Amblyomma variegatum ticks. In southern Africa, the principal vector is A. hebraeum. Febrile disease is a serious issue in the study area. There is a high prevalence of non-malaria illness caused by Rickettsia, so there is a need to have more knowledge on these species. Infection rates and transovarial transmission efficiency of R. africae in A. hebraeum ticks were investigated in a rural area of Mpumalanga province, South Africa. Adult and engorged A. hebraeum female ticks were collected from cattle. Larvae were collected by dragging a cloth at ground level using 100 steps, equivalent to an area of 100 m². Tick identification was performed according to standard taxonomic keys using a microscope. Engorged ticks were incubated to oviposit and egg masses were collected. DNA was extracted from the ticks, larvae and egg masses, and screened for gltA and ompA genes, using quantitative real-time PCR and conventional PCR, respectively. Positive ompA amplicons were sequenced and phylogenetic analysis showed 99.8-100% identity with R. africae. Infection rates were 13.7 and 12.7% for adults and larvae, respectively. Transovarial transmission of R. africae in A. hebraeum from this study was 85.7%. The results provide a clear indication that people living in the study area and travellers that visit the area are at risk of contracting ATBF.

Rickettsia africae is a bacterium of zoonotic importance, which causes African tick bite fever (ATBF) in humans. This pathogen is transmitted by ticks of the genus Amblyomma, with Amblyomma hebraeum and Amblyomma variegatum being the major vectors. Tick species other than the above-mentioned have also been reported to carry R. africae DNA. There is scarcity of information on the epidemiology of this pathogen, yet several cases have been recorded in foreign travellers who visited endemic areas, especially southern Africa. The disease has rarely been described in people from endemic regions. The aim of this study was to discuss the information that is currently available on the epidemiology of R. africae, highlighting the gaps in this field. Furthermore, ATBF cases, clinical signs and the locations where the cases occurred are also listed in this review.