Investigations of the Theileria parva carrier-state in cattle at the livestock/wildlife interface of the uPhongolo-Mkuze area in KwaZulu Natal, South Africa

Corridor disease (Theileria parva infection in cattle associated with carrier buffaloes) was not reported to cause serious outbreaks prior to 1994. From 2002-2004, outbreaks in cattle have increased in the areas where the disease is endemic in buffalo populations. In this study, the occurrence of Corridor disease outbreaks in the Zululand district municipality was closely monitored from 2004-2009. The observations included the number of cattle involved in the outbreaks, clinical signs, parasitological and post-mortem examinations while blood for serum and in EDTA were collected for serological (IFA test) and molecular (real-time PCR) tests specific for T. parva. Samples were collected from cattle involved in the outbreak, the sick and presumed recovered cattle. Recovered cattle from the farms were brought to the laboratory at the Onderstepoort Veterinary Institute for further investigations. This included tick pick-up and transmission attempts to demonstrate their carrier status as well as assessing their immunity to further experimental challenge using virulent T. parva stabilate. Results were obtained on Corridor disease outbreaks in the study area and ad hoc locations comprising a total of 15 commercial farms and community diptanks in the district from 2004 to 2009. A total of 31 outbreaks were recorded during the study period. The number of outbreaks per year was stable, being 3 or 4 from 2004 to 2007. A 100 percent increase was recorded in the subsequent years, 2008-2009. In one location, Morgenzon farm comprising a commercial and community farmers, had experienced regular outbreaks from 2004-2009. It is also noted that some farms experienced outbreaks for three consecutive years. Three other farms had experienced outbreaks for the first time in either 2008 or 2009. The most severe outbreak occurred in Nyalisa in 2009 where the disease was experienced for the first time in one herd in which 202 cattle were involved and 57 died within 30-40 days after the onset of the disease. Using all the tools mentioned above, the cause of death was confirmed to be due to T. parva infection. The Corridor disease outbreaks that were investigated, have mostly been reported during the months from March-May (88 %) but some (8 %) were encountered during the winter months (June-August). The distribution of outbreaks mainly coincided with the activity period of adult R. appendiculatus. During the investigation period, a total of 846 cattle were tested for Corridor disease and the prevalence was found to be 27 %. The percentage of cattle which were found positive by PCR was 16.5. Seven percent were found positive on both PCR and IFA tests, an indication of the development of a carrier state. However, 10 % of the cattle remained sero-positive with no indication of being parasite-carriers (real-time PCR negative). Five cattle which recovered from an apparent severe T. parva infection in the field and confirmed to be positive by PCR, all became negative before they were used in the transmission experiments. Ticks derived from these cattle were used to infect susceptible bovines but only T. taurotragi was transmitted. The xeno-diagnosis failed to demonstrate the carrier state in these field cattle. The five Corridor disease recovered cattle obtained from different study locations mentioned above, received lethal challenge using T. parva buffalo-derived stabilate. All challenged animals, including the susceptible control, showed schizont parasitosis as detected by the T. parva</i. real-time PCR test starting day 11 to 23. All animals also developed significant antibody titer to T. parva by day 28. Of the field cattle, only one bovine which showed mild reactions manifested by high temperature on day 11 for two consecutive days and schizonts parasitosis in lymph nodes on day 15 for only two days and recovered. The rest of the field cattle did not show any clinical or parasitological reactions during the observation period (103 days). The control bovine had high fever and showed schizonts parasitosis by day 11 for seven consecutive days. The reaction was classified as severe and had to be treated. Unfed R. appendiculatus collected off grass from one of the study sites were applied to feed on a susceptible bovine and only T. taurotragi was transmitted. There were no apparent clinical signs and the animal behavior kept normal during the observation period (60 days). This study suggests that Corridor disease should be considered as an “emerging disease” and more stringent control methods should be implemented. CopyrightDissertation (MSc)--University of Pretoria, 2012.Veterinary Tropical Diseasesunrestricte

South African buffalo-derived Theileria parva is distinct from other buffalo and cattle-derived T. parva

Theileria parva is a protozoan parasite transmitted by the brown-eared ticks, Rhipicephalus appendiculatus and Rhipicephalus zambeziensis. Buffaloes are the parasite’s ancestral host, with cattle being the most recent host. The parasite has two transmission modes namely, cattle–cattle and buffalo–cattle transmission. Cattle–cattle T. parva transmission causes East Coast fever (ECF) and January disease syndromes. Buffalo to cattle transmission causes Corridor disease. Knowledge on the genetic diversity of South African T. parva populations will assist in determining its origin, evolution and identify any cattle–cattle transmitted strains. To achieve this, genomic DNA of blood and in vitro culture material infected with South African isolates (8160, 8301, 8200, 9620, 9656, 9679, Johnston, KNP2, HL3, KNP102, 9574, and 9581) were extracted and paired-end whole genome sequencing using Illumina HiSeq 2500 was performed. East and southern African sample data (Chitongo Z2, Katete B2, Kiambu Z464/C12, Mandali Z22H10, Entebbe, Nyakizu, Katumba, Buffalo LAWR, and Buffalo Z5E5) was also added for comparative purposes. Data was analyzed using BWA and SAMtools variant calling with the T. parva Muguga genome sequence used as a reference. Buffalo-derived strains had higher genetic diversity, with twice the number of variants compared to cattle-derived strains, confirming that buffaloes are ancestral reservoir hosts of T. parva. Host specific SNPs, however, could not be identified among the selected 74 gene sequences. Phylogenetically, strains tended to cluster by host with South African buffalo-derived strains clustering with buffalo-derived strains. Among the buffalo-derived strains, South African strains were genetically divergent from other buffalo-derived strains indicating possible geographic sub-structuring. Geographic substructuring was also observed within South Africa strains. The knowledge generated from this study indicates that to date, ECF is not circulating in buffalo from South Africa. It also shows that T. parva has historically been present in buffalo from South Africa before the introduction of ECF and was not introduced into buffalo during the ECF epidemic.The Department of Agriculture, Land Reform and Rural Developmenthttp://www.frontiersin.org/Geneticsam2022Veterinary Tropical Disease

Safety and efficacy of an attenuated heartwater (Ehrlichia ruminantium) vaccine administered by the intramuscular route in cattle, sheep and Angora goats

Please read abstract in the article.Red Meat Research and Development Trust of South Africa; Mohair South Africa LTD; Technology Innovation Agency, South Africa; Agricultural Research Council, South Africa.http://www.elsevier.com/locate/vaccinehj2021Veterinary Tropical Disease

Molecular genotyping and epidemiology of equine piroplasmids in South Africa

Recently reported substantial genetic diversity within Theileria equi 18S rRNA gene sequences has led to the identification of five genotypes A, B, C, D, and E, complicating molecular and serological diagnosis. In addition, T. haneyi has lately been reported as a species closely related to the T. equi 18S rRNA genotype C (Knowles et al., 2018). Theileria spp. of this group have a monophyletic origin and are therefore referred to as Equus group to distinguish them from the remaining Theileria lineages (Jalovecka et al., 2019). In this study, we report on the development of genotype-specific quantitative real-time PCR assays capable of detecting and distinguishing between each parasite genotype. Alignment of complete 18S rRNA sequences available on GenBank allowed for the design of a single primer pair and five TaqMan minor groove binder (MGB™) probes specific for each genotype (A–E). The assays, evaluated as qPCR simplex and two qPCR multiplex formats (Multiplex EP–ABC and Multiplex EP–DE), were shown to be both efficient and specific in the detection of T. equi genotypes. The developed qPCR assays were used to study (i) the intra-specific diversity of parasite genotypes within horse and zebra, (ii) the inter-specific differences in parasite genotype diversity in horses as compared to zebra, and (iii) the geographic distribution of T. equi 18S rRNA genotypes in South Africa. In addition, (iv) the presence of T. haneyi in South Africa was evaluated. An assessment of 342 equine field samples comprising 149 field horses, 55 racehorses, and 138 wild zebra confirmed the previously reported presence of T. equi 18S rRNA genotypes A, B, C, and D, and absence of genotype E in South African equids. Theileria equi genotypes A, B, C, and D, were detected in zebra, whereas only genotypes A, C and D, could be identified in field horses, and only genotypes A and C in racehorses. Genotypes B and D were the dominant genotypes identified in zebra in South Africa, while horses were predominantly infected with T. equi genotypes A and C. The greater diversity of T. equi genotypes in zebra suggests that it is an ancestral host for this piroplasmid lineage. Importantly, evidence is presented that each identified T. equi genotype segregates independently in each of the three studied equid populations reinforcing the notion that they represent individual separate entities corresponding to species. Preliminary investigations of the relationship between T. equi genotype C infections and Theileria haneyi, suggest that in addition to the five currently known T. equi genotypes, South African equids are also infected with T. haneyi.Fil: Bhoora, Raksha Vasantrai. University of Pretoria; Sudáfrica. Agricultural Research Council. Onderstepoort Veterinary Research; SudáfricaFil: Collins, Nicola Elaine. University of Pretoria; SudáfricaFil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología Veterinaria - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Patobiología Veterinaria; ArgentinaFil: Troskie, Christo. Agricultural Research Council. Onderstepoort Veterinary Research; SudáfricaFil: Marumo, Ratselane. Agricultural Research Council. Onderstepoort Veterinary Research; SudáfricaFil: Labuschagne, Karien. Agricultural Research Council. Onderstepoort Veterinary Research; SudáfricaFil: Smith, Rae Marvin. South African National Biodiversity Institute; SudáfricaFil: Dalton, Desire Lee. South African National Biodiversity Institute; Sudáfrica. University of Venda; SudáfricaFil: Mbizeni, Sikhumbuzo. Agricultural Research Council. Onderstepoort Veterinary Research; Sudáfrica. University of South Africa; Sudáfric