Molecular detection of Coxiella burnetii and Coxiella species in rats and chickens from poultry farms in North West Province, South Africa.

BACKGROUND: Coxiella burnetii is a bacterial pathogen that causes query fever and coxiellosis in humans and animals, respectively. There is a scarcity of studies on the prevalence of C. burnetii infections in rats and chickens in South Africa. OBJECTIVE: The aim of this study was to determine the occurrence of C. burnetii in rats and chickens sampled from poultry farms in the North West Province of South Africa. METHODS: DNA was extracted from rodent kidneys (n = 68) and chicken faeces (n = 52). Two rodent pest species, namely Rattus rattus and Rattus tanezumi, were identified by analysis of CO1 gene sequences. Detection of C. burnetii was carried out using polymerase chain reaction assays targeting 23S rRNA, 16S rRNA and IS111 markers. RESULTS: C. burnetii was detected in 16.2%, 8.8% and 25% of R. rattus, R. tanezumi and chickens, respectively. CONCLUSIONS: The findings in this study demonstrate that rodents and chickens are harbouring C. burnetii at sampled poultry farms. There should be frequent screening for C. burnetii in poultry operations. The likelihood of future transmission between rodents and chickens, including humans, also needs to be investigated.

First report of gastrointestinal nematodes and coccidia parasites from free-range chickens in Mafeteng district, Lesotho.

Free-range chickens are an integral part of poultry production in many developing countries. In the Mountain Kingdom of Lesotho, the majority of the population own free-range chickens, which serve a variety of purposes including being a source of meat, eggs and use for cultural rituals amongst others. However, there is lack of scientific studies on occurrence of parasitic infections on free-range chickens in Lesotho. The aim of this study was to document common gastrointestinal parasites infecting free-range chickens in four villages of Mafeteng District in Lesotho. A total number of 462 pooled faecal samples were collected from various households in HaKubutu (n = 114), HaMatjeka (n = 120), HaMpalipali (n = 120) and Thabang Villages (n = 108) which were subjected to microscopic examination using McMaster technique. The prevalence of gastrointestinal parasite infection was as follows: Eimeria tenella (12.8%), Ascaridia galli (10.4%) and Heterakis gallinarum (5%). The prevalence for H. gallinarum and Ascaridia galli were comparatively higher during the hot-wet season (7.1% and 2.8% respectively) than the cold-dry season (3.2% and 1.9% respectively) and varied significantly (P < 0.05). For E. tenella, the oocysts per gram were slightly higher in the cold-dry season than the hot-wet season. Polymerase chain reaction only amplified DNA from six (29%) adult A. galli worms of which two amplicons were successfully sequenced. The obtained cytochrome C oxidase subunit 1 partial gene sequences displayed 98-100% identity with South African A. galli isolates. This is the first scientific study on prevalence and molecular characterization of nematodes and coccidia species infecting free-range village chickens in Lesotho. The findings can be used to review management of gastrointestinal nematodes and protozoal parasites of free-range chickens in Lesotho.

Detection of Ticks and Tick-Borne Pathogens of Urban Stray Dogs in South Africa.

This study aimed to identify ticks infesting dogs admitted to the Potchefstroom Animal Welfare Society (PAWS) and to detect tick-borne pathogens they are harbouring. A total of 592 ticks were collected from 61 stray dogs admitted to PAWS originating from several suburbs in and near Potchefstroom, South Africa. The dog ticks were identified as Haemaphysalis elliptica (39%) and Rhipicephalus sanguineus (61%) by both morphological and DNA analyses. Of these ticks, H. elliptica consisted of 67.5% (156/231) and 32.5% (75/231) female and male ticks, respectively, whilst R. sanguineus consisted of 48.5% (175/361) and 51.5% (186/361) female and male ticks, respectively. Microscopic examination of blood smears from engorged female ticks indicated overall occurrences of 0.5% (1/204) for Babesia spp. from R. sanguineus, 1% (2/204) of Anaplasma spp. from H. elliptica, and 22% (45/204) of Rickettsia spp. from both H. elliptica and R. sanguineus. Using pooled samples molecular detection of tick-borne pathogens indicated overall occurrences of 1% (1/104) for A. phagocytophilum in H. elliptica, 9.6% (10/104) of Rickettsia spp. in H. elliptica and R. sanguineus, 5.8% (6/104) of Ehrlichia canis in H. elliptica and R. sanguineus, and 13.5% (14/104) of Coxiella spp. in both H. elliptica and R. sanguineus. Additionally, PCR detected 6.5% (2/31) of Coxiella spp. DNA from H. elliptica eggs. Our data indicate that urban stray dogs admitted at PAWS are infested by H. elliptica and R. sanguineus ticks which are harbouring several pathogenic organisms known to cause tick-borne diseases.

The COMBAT project: controlling and progressively minimizing the burden of vector-borne animal trypanosomosis in Africa.

Vector-borne diseases affecting livestock have serious impacts in Africa. Trypanosomosis is caused by parasites transmitted by tsetse flies and other blood-sucking Diptera. The animal form of the disease is a scourge for African livestock keepers, is already present in Latin America and Asia, and has the potential to spread further. A human form of the disease also exists, known as human African trypanosomosis or sleeping sickness. Controlling and progressively minimizing the burden of animal trypanosomosis (COMBAT) is a four-year research and innovation project funded by the European Commission, whose ultimate goal is to reduce the burden of animal trypanosomosis (AT) in Africa. The project builds on the progressive control pathway (PCP), a risk-based, step-wise approach to disease reduction or elimination. COMBAT will strengthen AT control and prevention by improving basic knowledge of AT, developing innovative control tools, reinforcing surveillance, rationalizing control strategies, building capacity, and raising awareness. Knowledge gaps on disease epidemiology, vector ecology and competence, and biological aspects of trypanotolerant livestock will be addressed. Environmentally friendly vector control technologies and more effective and adapted diagnostic tools will be developed. Surveillance will be enhanced by developing information systems, strengthening reporting, and mapping and modelling disease risk in Africa and beyond. The socio-economic burden of AT will be assessed at a range of geographical scales. Guidelines for the PCP and harmonized national control strategies and roadmaps will be developed. Gender equality and ethics will be pivotal in all project activities. The COMBAT project benefits from the expertise of African and European research institutions, national veterinary authorities, and international organizations. The project consortium comprises 21 participants, including a geographically balanced representation from 13 African countries, and it will engage a larger number of AT-affected countries through regional initiatives.

Bacterial pathogens identified from houseflies in different human and animal settings: A systematic review and meta-analysis.

Housefly (Musca domestica) is an excellent candidate for the distribution of susceptible and resistant bacterial strains that potentially threaten public health. To date, there is a paucity of information on the global distribution of pathogenic bacteria of medical and veterinary importance from diverse environmental settings. Therefore, this study was undertaken to conduct a systemic review and meta-analysis to estimate occurrence of various bacterial species of medical and veterinary importance harboured by houseflies around the world. Published articles from 1980 to 2020 were retrieved from electronic databases and assessed for eligibility according to Preferred Reporting Items for Systemic Reviews and Meta-Analysis guidelines. Seventy-eight studies were included in the review with only 44 studies being eligible for meta-analysis. Results indicated that eligible studies used in this review were from four continents, i.e., Asia (47.4%) America (23.1%), Africa (20.5%) and Europe (8.9%). The majority of the studies (56.4%) used the culture method for the identification of bacterial pathogens, while 30.7% used both culture and PCR techniques. For meta-analysis, we focused on five pathogenic bacterial species including Escherichia coli, Enterococcus faecium, Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus. High heterogeneity was found among studies investigating different pathogens including E. coli (Q = 10,739.55; I2  = 99.60; Q-p 0.0001), E. faecium (Q = 317.61; I2  = 86.46; Q-p < 0.0001), K. pneumonia (Q = 1,576.61; I2  = 97.27; Q-p < 0.0001), S. aureus (Q = 2,439.12; I2  = 98.24; Q-p < 0.0001) and P. aeruginosa (Q = 1,283.0; I2  = 96.65; Q-p < 0.0001). Furthermore, it was observed that houseflies carried a considerable number of susceptible and antibiotic-resistant bacterial strains that pose considerable threats to public health. Findings from this study have provided more insight on the vectoral potential of houseflies in the transmission of significant bacterial pathogens from different regions across the world. Further investigation is required to quantify the bacterial contamination and dissemination by houseflies.

A distribution model for Glossina brevipalpis and Glossina austeni in Southern Mozambique, Eswatini and South Africa for enhanced area-wide integrated pest management approaches.

BACKGROUND: Glossina austeni and Glossina brevipalpis (Diptera: Glossinidae) are the sole cyclical vectors of African trypanosomes in South Africa, Eswatini and southern Mozambique. These populations represent the southernmost distribution of tsetse flies on the African continent. Accurate knowledge of infested areas is a prerequisite to develop and implement efficient and cost-effective control strategies, and distribution models may reduce large-scale, extensive entomological surveys that are time consuming and expensive. The objective was to develop a MaxEnt species distribution model and habitat suitability maps for the southern tsetse belt of South Africa, Eswatini and southern Mozambique. METHODOLOGY/PRINCIPAL FINDINGS: The present study used existing entomological survey data of G. austeni and G. brevipalpis to develop a MaxEnt species distribution model and habitat suitability maps. Distribution models and a checkerboard analysis indicated an overlapping presence of the two species and the most suitable habitat for both species were protected areas and the coastal strip in KwaZulu-Natal Province, South Africa and Maputo Province, Mozambique. The predicted presence extents, to a small degree, into communal farming areas adjacent to the protected areas and coastline, especially in the Matutuíne District of Mozambique. The quality of the MaxEnt model was assessed using an independent data set and indicated good performance with high predictive power (AUC > 0.80 for both species). CONCLUSIONS/SIGNIFICANCE: The models indicated that cattle density, land surface temperature and protected areas, in relation with vegetation are the main factors contributing to the distribution of the two tsetse species in the area. Changes in the climate, agricultural practices and land-use have had a significant and rapid impact on tsetse abundance in the area. The model predicted low habitat suitability in the Gaza and Inhambane Provinces of Mozambique, i.e., the area north of the Matutuíne District. This might indicate that the southern tsetse population is isolated from the main tsetse belt in the north of Mozambique. The updated distribution models will be useful for planning tsetse and trypanosomosis interventions in the area.

Application of culture, PCR, and PacBio sequencing for determination of microbial composition of milk from subclinical mastitis dairy cows of smallholder farms.

Mastitis is a cow disease usually signalized by irritation, swelling, and soreness of the udder. It is characterized by physical, chemical, and biological changes in the udder and milk. The aim of this study was to detect and characterize pathogens causing subclinical mastitis (SCM) from the milk of dairy cows of small-scale farmers through culture and molecular techniques. Milk was collected from 32 cows belonging to 8 small-scale farmers around Harrismith District, South Africa. The results showed that screening of SCM by California mastitis test and somatic cell counts (SCC) was 21.87 and 25%, respectively. Culture methods revealed the presence of Staphylococcus aureus at 93% followed by Streptococci spp. and Escherichia coli at 36.4 and 13.3%, respectively. The PCR could only detect E. coli, while single-molecule real-time sequencing showed a total of 2 phyla, 5 families, 7 genera, and 131 species. Clostridiaceae was the most abundant family, while Romboutsia was the most abundant genus followed by Turicibacter spp. The present study has documented the occurrence of SCM causing pathogens in milk collected from cows of small-scale farmers in Harrismith, indicating that SCM may be present at higher levels than expected.

Mosquito identification and haemosporidian parasites detection in the enclosure of the African penguins (Spheniscus demersus) at the SANBI zoological garden.

The National Zoological Gardens (NZG) is a facility of the South African National Biodiversity Institute (SANBI) and the largest zoo in southern Africa. Among the 9000 captive animals kept by the NZG, is the endangered African penguin (Spheniscus demersus). There have been several post-mortem reports on deaths of penguins in the NZG due to haemosporidian infections, however, the haemosporidian lineages involved and possible insect vector are unknown. Haemosporidians are apicomplexan parasites that infect vertebrates through blood-sucking dipteran insects. Therefore, the current study aimed to identify mosquitoes that are potential vectors found within the African penguin enclosure as well as to detect the haemosporidian parasites from these insects using nested-PCR and real-time PCR (qPCR) analyses. Mosquito samples were collected using an overnight UV-light trap setup for 3 months. From the 65 pooled samples representing 325 mosquitoes, morphological and molecular analysis showed that Culex pipiens (52.31%) was the dominant species followed by Cx. t heileri (30.77%) and Cx. quinquefasciatus (16.92%). Nested-PCR detected parasite DNA of Leucocytozoon sp. and Plasmodium sp. The Cx. pipiens had the highest minimum infection rate (MIR) of 5.88% by nested-PCR and 9.41% by qPCR whilst Cx. quinquefasciatus had MIR of 3.64% in both assays and no haemosporidian parasites were detected from Cx. t heileri. One Cx. pipiens sample had a co-infection of both Plasmodium sp. and Leucocytozoon sp. detected by nested-PCR. These findings suggest that effective control measures for blood-sucking dipteran insects is required at the NZG and more studies should be conducted to determine the actual prevalence of these haemosporidian parasites among other bird species within NZG.

Molecular detection of virulence genes in Salmonella spp. isolated from chicken faeces in Mafikeng, South Africa.

Chickens have been implicated in most Salmonella disease outbreaks because they act as carriers of the pathogen in their gut. There are over 2500 serotypes of Salmonella that have been reported worldwide and 2000 of these serovars can be found in chickens. The main objective of this study was to determine the Salmonella serotypes found in poultry farms around Mafikeng district, South Africa. Salmonella was identified according to the guidelines of the International Organization for Standardization (ISO) (ISO 6579:2002) standard techniques. Faecal samples were collected and analysed for Salmonella using conventional cultural methods and polymerase chain reaction targeting the 16S Ribosomal Deoxyribonucleic acid (rDNA) gene for Salmonella identification. Out of 130 presumptive Salmonella isolates determined by urease and triple sugar iron tests, only 46 isolates were identified as Salmonella serotypes of which S. Typhimurium was the most frequent with 18 (39.1%), followed by S. Heidelberg with 9 (19.6%), S. bongori with 7 (15.2%), S. Enteritidis with 6 (13.0%) and both S. Paratyphi B and S. Newport with 3 (6.5%) each. Seven virulence genes including invA 100%, spy 39%, hilA 9%, misL 30%, sdfI 13%, orfL 11% and spiC 9% were detected from these Salmonella isolates in this study. The presence of these virulence genes indicates high pathogenicity potential of these isolates which is a serious public health concern because of zoonotic potential of Salmonella.

Equine piroplasmosis: an insight into global exposure of equids from 1990 to 2019 by systematic review and meta-analysis.

Equine piroplasmosis (EP) is a tick-borne disease of economic importance, relevant in the international movement of equids. The causative agents are at least two apicomplexan protozoan parasites Babesia caballi and Theileria equi. To date, there is no study that estimates global and regional exposure of equids to EP. We therefore conducted a systematic review and meta-analysis to estimate the pooled prevalence and heterogeneity of EP using random-effects model. Six electronic databases were searched for publications on EP and assessed according to Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. A total of 66 eligible studies published between 1990 and 2019 and representing 24 041 equids were included. The overall pooled prevalence estimates (PPEs) of B. caballi was 22.3% (95% CI 21.7-22.8), while the overall PPE for T. equi was 29.4% (95% CI 28.7-30.0). The overall pooled prevalence due to co-infection with both parasites was 11.8% (95% CI 11.32-12.32). Also, subgroup analysis according to sex, age, diagnostic technique, equid species, region and publication years showed a substantial degree of heterogeneity across studies computed for both B. caballi and T. equi infections in equids. Awareness of the current status of EP globally will alert the relevant authorities and stakeholders where necessary on the need for better preventive and control strategies against the disease.

Molecular evidence of Babesia caballi and Theileria equi in equines and ticks in Nigeria: prevalence and risk factors analysis.

Babesia caballi and Theileria equi are biological agents responsible for equine piroplasmosis (EP). We conducted a robust and extensive epidemiological study in Nigeria on the prevalence and risk factors of EP. Blood (468, both horses and donkeys) and ticks (201 pools) were screened using polymerase chain reaction (PCR). DNA of equine piroplasms was observed in tick pools with B. caballi amplified in Rhipicephalus evertsi evertsi only [minimum infection rate (MIR) of 7.6%] while T. equi was observed in R. e. evertsi (MIR, 61.6%), Hyalomma dromedarii (MIR, 23.7%) and H. truncatum (MIR, 50.0%). Overall results showed that 196/468 (41.9%) animals were positive for equine piroplasms (both B. caballi and T. equi). The prevalence for T. equi was 189/468 (40.4%) compared to 7/468 (1.5%) for B. caballi. In the horses and donkeys, respectively, the prevalence for T. equi was (39.9%; 112/281) and (41.2%; 77/187) compared with (1.4%; 4/281) and (1.6%; 3/187) due to B. caballi. Our analysis showed that location (Jigawa state), Talon breed, horses used for work and reproduction, unsatisfactory husbandry practices, contact with other mammals are risk factors that associated positivity to T. equi infection in horses, whilst horses kept on intensive management appeared to be less prone to infection. On the other hand, Jangora breed of donkeys and location (Jigawa state) are risk factors to infection with T. equi in donkeys. Findings suggest the persistence of EP in equids and ticks in Nigeria.

Characterization of tabanid flies (Diptera: Tabanidae) in South Africa and Zambia and detection of protozoan parasites they are harbouring.

Tabanids are haematophagous flies feeding on livestock and wildlife. In the absence of information on the relationship of tabanid flies and protozoan parasites in South Africa and Zambia, the current study was aimed at characterizing tabanid flies collected in these two countries as well as detecting protozoan parasites they are harbouring. A total of 527 tabanid flies were collected whereby 70·2% were from South Africa and 29·8% were from Zambia. Morphological analysis revealed a total of five different genera collected from the sampled areas namely: Ancala, Atylotus, Haematopota, Philoliche and Tabanus. DNA extracted from South African Tabanus par and Tabanus taeniola tested positive for the presence of Trypanosoma congolense (Savannah) and Trypanosoma theileri whilst one member from T. par was positive for Trypanosoma brucei species. DNA extracted from Zambian tabanid flies tested positive for the presence of Besnoitia species at 1·27% (2/157), Babesia bigemina 5·73% (9/157), Theileria parva 30·11% (30/157) and 9·82% (14/157) for Trypanosoma evansi. This study is the first to report on relationship of Babesia and Theileria parasites with tabanid flies. Further investigations are required to determine the role of tabanids in transmission of the detected protozoan parasites in livestock and wildlife in South Africa and Zambia.