Genetic testing of canine degenerative myelopathy in the South African Boxer dog population

Canine degenerative myelopathy (DM) is a progressive disease process that is diagnosed late in life and mainly affects the pelvic limbs. Factors that make an ante-mortem definitive diagnosis of DM include: an insidious onset and clinical manifestation that mimics other disease processes of the pelvic limbs (hip dysplasia, cranial cruciate ligament rupture, etc.) or there may even be concurrent disease processes, old-age onset and lack of reliable diagnostic methods. Until recently, South African dog owners had to submit samples to laboratories overseas for genetic testing in order to confirm an affected dog (homozygous A/A) and to aid in the ante-mortem diagnosis of DM. Only affected dogs have been confirmed to manifest the clinical signs of DM. This study aimed to verify whether genetic testing by a local genetic laboratory was possible in order to detect a missense mutation of the superoxide dismutase gene (SOD1) that is implicated in causing the clinical signs of DM. The study also aimed to detect and map the inheritance of this disease process in a local Boxer dog population where the pedigree of the sampled population was known. Venous blood collected from Boxer dogs using a simple random sampling technique. The samples were genotyped for the SOD1:c.118G>A polymorphism. Carrier and affected Boxer dogs were detected. A pedigree that demonstrated the significance of inheriting a carrier or affected state in the population was mapped. The present study concludes that genotyping of the missense mutation in Boxer dogs is possible in South Africa. There are carrier and affected Boxer dogs in the local population, making DM a plausible diagnosis in aged dogs presenting with pelvic limb pathology

Babesia species of domestic cats : molecular characterization has opened Pandora's box

This is the first comprehensive review of the literature pertaining to Babesia species reported from domestic cats. Description of the four species (Babesia felis, Babesia cati, Babesia herpailuri, and Babesia pantherae) named based on morphology and/or host specificity is documented. Feline babesiosis is of major veterinary concern only in South Africa. Reports of the rare occurrence of feline babesiosis cases in Europe (France, Germany, Poland, and Spain) and Asia (Israel, India, and Pakistan) are documented. Molecular characterization has revealed that cats can harbor a variety of Babesia species. The previous practice of referring to all piroplasms, especially small ones, seen on feline blood smears as B. felis is therefore no longer tenable. The near-full-length 18S rRNA gene sequences entered into GenBank in 2001 (accession no. AF244912) are designated as definitive for B. felis sensu stricto. All published literature relating to molecular characterization of feline Babesia species that could be traced was critically assessed. Four Babesia species are now known to be involved in causing feline babesiosis in South Africa: the closely related B. felis s.s. and Babesia leo (clade I), Babesia lengau (clade II), and Babesia species cat Western Cape (clade VI, Babesia s.s.). Clade VI also includes Babesia canis presentii and Babesia hongkongensis reported from cats in Asia. Six other Babesia species have been reported from domestic cats: the dog-associated B. canis s.s., Babesia gibsoni, and B. vogeli, as well as Babesia lohae, Babesia microti, and Babesia vulpes. Phylogenetic relationships of all named species were assessed and are presented as trees. The relatively high prevalence of B. vogeli in clinically healthy cats (16%in Brazil, 13%on St Kitts, and 8.1%in Portugal) suggests that immunocompetent cats can harbor the infection with no discernible untoward effects. Reports of occurrence of B. felis and other Babesia species in domestic cats should be accepted only if they are supported by credible molecular provenance.https://www.frontiersin.org/journals/veterinary-science#am2020Veterinary Tropical Disease

Development and validation of a TaqMan® probe-based real-time PCR assay for detection of Ehrlichia canis

Ehrlichiosis is a potentially fatal zoonotic tick-borne disease, caused by a pleomorphic Gram-negative bacterium. It occurs worldwide and affects humans, domestic and wild animals. Dogs infected with Ehrlichia canis develop canine monocytic ehrlichiosis (CME), a significant infectious disease of canines. TaqMan® based real-time PCR assays to detect Ehrlichia spp. affecting dogs were developed and a real-time PCR assay specific for E. canis validated. The efficiency of the assay was 93% and the 95% limit of detection was 33 E. canis plasmid copies/µl of blood (95% confidence interval: 23 - 58). The assay was specific for E. canis when tested against other haemoparasites. Consistent repeatability was observed, with an inter-run standard deviation (SD) range between 0.33 and 1.29 and an intra-run SD range between 0.04 and 1.14. Field samples were tested in parallel by both the E. canis real-time PCR assay and a reverse line blot hybridization assay. The results were in agreement for the two assays, with an exception of two out of 121 samples. Bayesian latent class analysis was used to calculate a diagnostic sensitivity of the E. canis real-time PCR assay of 90% and a specificity of 92%. This assay is a sensitive and reliable molecular detection method for E. canis and will be a useful tool for early diagnosis and timely treatment for this haemoparasite.The National Research Foundation (NRF-BAAP) and Agricultural Sector Education Training Authority (AgriSETA).https://www.elsevier.com/locate/ttbdishj2022Veterinary Tropical Disease

A One Health approach to investigate bats as a potential source of zoonotic mycoses in selected areas of Mpumalanga province, the Republic of South Africa

A One Health approach pilot study was carried out in selected villages within the Mnisi Traditional Authority’s area, Manyeleti Game Reserve, and Hans Hoheisen Wildlife Research Station in Mpumalanga Province, the Republic of South Africa from July to December 2018. The study’s main objectives were to identify positive and negative human-bat-environment interactions and microbiological screening of bats’ faecal samples for zoonotic fungi. Thirty-three purposively selected participants were asked to complete a structured questionnaire with multiple-choice and open-ended questions, and a total of 55 faecal samples were collected, 25 from identified bat roosting sites and 30 from captured bats. Ninety seven percent of respondents were aware of the presence or absence of bats in their immediate surroundings. However, the majority of them (87.9%) were uneasy about the presence of bats in their buildings, and nearly half (48.5%) were unsure whether bats play a positive or negative role in the environment. Some respondents (15.2%) stated that bats play beneficial roles in the environment, such as pollinating plants, spreading seeds of indigenous plants, catching harmful insects, and so on. More than half of the respondents (66.7%) stated that bats can be a nuisance; 18.2% of those polled reported contracting fungal diseases as a result of cleaning bat droppings without adequate protection. The analysis of faecal samples revealed that bats can harbour pathogenic fungi such as Aspergillus fumigatus, and A.flavus. We concluded that bats can harbour fungal pathogens that cause human diseases. Further research should be conducted to compile a complete list of fungi pathogens in bats in the study area

Phylogeny of Theileria buffeli genotypes identified in the South African buffalo (Syncerus caffer) population

Theileria buffeli/orientalis is a group of benign and mildly pathogenic species of cattle andbuffalo in various parts of the world. In a previous study, we identified T. buffeli in blood sam-ples originating from the African buffalo (Syncerus caffer) in the Hluhluwe–iMfolozi GamePark (HIP) and the Addo Elephant Game Park (AEGP) in South Africa. The aim of this studywas to characterise the 18S rRNA gene and complete internal transcribed spacer (ITS1-5.8S-ITS2) region of T. buffeli samples, and to establish the phylogenetic position of this speciesbased on these loci. The 18S rRNA gene and the complete ITS region were amplified fromDNA extracted from blood samples originating from buffalo in HIP and AEGP. The PCR prod-ucts were cloned and the resulting recombinants sequenced. We identified novel T. buffeli-like 18S rRNA and ITS genotypes from buffalo in the AEGP, and novel Theileria sinensis-like18S rRNA genotypes from buffalo in the HIP. Phylogenetic analyses indicated that the T.buffeli-like sequences were similar to T. buffeli sequences from cattle and buffalo in Chinaand India, and the T. sinensis-like sequences were similar to T. sinensis 18S rRNA sequencesof cattle and yak in China. There was extensive sequence variation between the novel T.buffeli genotypes of the African buffalo and previously described T. buffeli and T. sinensisgenotypes. The presence of organisms with T. buffeli-like and T. sinensis-like genotypes inthe African buffalo could be of significant importance, particularly to the cattle industry inSouth Africa as these animals might act as sources of infections to naïve cattle. This is thefirst report on the characterisation of the full-length 18S rRNA gene and ITS region of T.buffeli and T. sinensis genotypes in South Africa. Our study provides invaluable informationtowards the classification of this complex group of benign and mildly pathogenic species.South African National Research Foundation (NRFICD2006072000009) and UP Research Development Programme.http://www.elsevier.com/locate/vetparhb201

Piroplasm parasites of white rhinoceroses (Ceratotherium simum) in the Kruger National Park, and their relation to anaemia

As part of a larger survey to map the geographical distribution of Babesia and Theileria parasites in the southern African rhinoceros population, white rhinoceroses were sampled during routine immobilisations in the Kruger National Park. Polymerase chain reaction (PCR) and reverse line blot (RLB) hybridisation assays were used to screen for the presence of piroplasms and complete blood counts were used to assess associated changes in clinical parameters. Of the 195 rhinoceroses sampled, 71 (36.4 %) tested positive for the presence of Theileria bicornis, with no significant change in the haematological parameters measured, while 18 (9.2 %) tested positive for Theileria equi. None of the rhinoceroses sampled tested positive for Babesia bicornis, a parasite associated with mortalities in black rhinoceroses.Grant: National Research Foundation, Funding: Wildlife Group of the South African Veterinary Associationhttp://www.journals.co.za/ej/ejour_savet.htmlab201

Confirmation of occurrence of Babesia vogeli in a dog in Windhoek, central Namibia

Although there is evidence of high seroprevalence of antibodies to Babesia spp. in dogs in central Namibia, clinical babesiosis is rarely diagnosed. Rhipicephalus sanguineus sensu lato, the vector of Babesia vogeli, is common in Namibia while Haemaphysalis elliptica, the vector of the highly virulent but morphologically indistinguishable Babesia rossi, has rarely been recorded, mainly in northern Namibia. On the basis of vector occurrence, clinical cases of canine babesiosis in Windhoek, central Namibia, have been ascribed to B. vogeli. DNA extracted from a blood smear made from a sick dog was subjected to the reverse line blot hybridisation assay. The polymerase chain reaction amplicons hybridised with the B. vogeli–specific probe, but not with the Babesia canis– and B. rossi–specific probes. Although attempts at cloning and sequencing of the full-length 18S rRNA gene were unsuccessful, we can confirm that B. vogeli occurs in central Namibia.http://www.jsava.co.zaam2016Veterinary Tropical Disease

Sequence variation identified in the 18S rRNA gene of Theileria mutans and Theileria velifera from the African buffalo (Syncerus caffer)

The African buffalo (Syncerus caffer) is a natural reservoir host for both pathogenic and nonpathogenic Theileria species. These often occur naturally as mixed infections in buffalo. Although the benign and mildly pathogenic forms do not have any significant economic importance, their presence could complicate the interpretation of diagnostic test results aimed at the specific diagnosis of the pathogenic T. parva in cattle and buffalo in South Africa. The 18S rRNA gene has been used as the target in a quantitative real-time PCR (qPCR) assay for the detection of T. parva infections. However, the extent of sequence variation within this gene in the non-pathogenic Theileria spp. of the Africa buffalo is not well known. The aim of this study was, therefore, to characterize the full-length 18S rRNA genes of T. mutans, Theileria sp. (strain MSD) and T. velifera and to determine the possible influence of any sequence variation on the specific detection of T. parva using the 18S rRNA qPCR. The reverse line blot (RLB) hybridization assay was used to select samples which either tested positive for several different Theileria spp., or which hybridized only with the Babesia/Theileria genus-specific probe and not with any of the Babesia or Theileria speciesspecific probes. The full-length 18S rRNA genes from 14 samples, originating from 13 buffalo and one bovine from different localities in South Africa, were amplified, cloned and the resulting recombinants sequenced. Variations in the 18S rRNA gene sequences were identified in T. mutans, Theileria sp. (strain MSD) and T. velifera, with the greatest diversity observed amongst the T. mutans variants. This variation possibly explained why the RLB hybridization assay failed to detect T. mutans and T. velifera in some of the analysed samples.South African National Research Foundation (NRF ICD2006072000009) and UP Research Development Programme.http://www.elsevier.com/locate/vetparhb2013ab201

Histomorphology, ultrastructure and fatty acid composition of the adipose tissue in pansteatitis, the potentials in understanding the underlying mechanism and diagnosis of pansteatitis in the Nile crocodile

BACKGROUND : In an effort to characterize the fat body and other adipose tissue in the Nile crocodile and the effects of pansteatitis on the structure and composition of the adipose tissue, we evaluated the regional variation in structure and fatty acid composition of healthy farmed crocodiles and those affected by pansteatitis. METHODS : Adipose tissue samples were collected from the subcutaneous, visceral and intramuscular fat and the abdominal fat body of ten 4-year old juvenile crocodiles from Izinthaba Crocodile Farm, Pretoria, South Africa while pansteatitis samples were collected from visceral and intramuscular fat of crocodiles that had died of pansteatitis at the Olifant River, Mpumalanga, also in South Africa. Histomorphology, ultrastrustucture and fatty acid composition by fatty acid methyl ester (FAME) analysis were conducted. RESULTS : Histological examination showed regional variations in the adipose tissue especially in the collagen content of the ECM, tissue perfusion and division into lobes and lobules by fibrous capsule. Considerable fibrosis, mononuclear cell infiltration especially by macrophages and lymphocytes and toxic changes in the nucleus were observed in the pansteatitis samples. Regional variation in lipid composition especially in Myristoleic (C14:1), Erucic acid (C22:1n9), and Docosadienoic acid (C22:2n6) was observed. Most of the saturated and trans fatty acids were found in significant quantities in the pansteatitis samples, but had very low levels of the cis fatty acid and the essential fatty acids with C18 backbone. CONCLUSION : This study demonstrates that there exists some regional variation in histomorphology and fatty acid composition in the healthy adipose tissue of the Nile crocodile. It also showed that pansteatitis in the Nile crocodile might have been triggered by sudden change in energy balance from consumption of dead fish; and probable exposure to toxic environmental conditions with the evidence of up scaled monounsaturated long chain fatty acids composition and toxic changes in the leucocytes observed in pansteatitis in the present study.http://www.sherpa.ac.uk/romeo/issn/1476-511X/am2017Anatomy and PhysiologyParaclinical SciencesVeterinary Tropical Disease

Black-backed jackals (Canis mesomelas) are natural hosts of Babesia rossi, the virulent causative agent of canine babesiosis in sub-Saharan Africa

BACKGROUND : Babesia rossi, which is transmitted by Haemaphysalis spp. and is highly virulent to domestic dogs, occurs only in sub-Saharan Africa. Since dogs are not native to the region, it has been postulated that the natural host of B. rossi is an indigenous African canid. Although various attempts at artificial infection indicated that black-backed jackals (Canis mesomelas) could become subclinically infected with B. rossi, data on occurrence of B. rossi in free-ranging jackals was lacking. A long-term behaviour study in which free-ranging black-backed jackals were radio-collared offered the opportunity of collecting blood specimens from a large number of free-ranging jackals. METHODS : Genomic DNA was extracted from the EDTA blood samples (n = 107). PCR products were subjected to Reverse Line Blot hybridization using Theileria and Babesia genera-specific as well as 28 species-specific oligonucleotide probes, including Babesia canis, Babesia rossi, Babesia vogeli and Babesia gibsoni. The near full-length parasite 18S rRNA gene was amplified from two selected samples (free-ranging jackals), cloned and a total of six recombinants were sequenced. RESULTS : Of 91 free-ranging jackals, 77 (84.6%) reacted with the Babesia genus-specific probe; 27 (29.7%) also reacted with the B. rossi probe. Of 16 captive jackals, 6 (37.5%) reacted with the B. rossi probe, while one further sample reacted with the Babesia genus-specific probe only. After cloning, 6 recombinants yielded identical sequences identical to that of B. rossi (L19079) and differing by 2 base pairs from B. rossi (DQ111760) in GenBank. The observed sequence similarities were confirmed by phylogenetic analyses using neighbour joining and maximum parsimony. CONCLUSIONS : Black-backed jackals are natural hosts of B. rossi.Laboratory expenses were funded by the Foundational Biodiversity Information Programme, National Research Foundation of South Africa (Grant 98110 to BLP).http://www.parasitesandvectors.comam2017Veterinary Tropical Disease