Molecular detection of Shiga toxin and extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolates from sheep and goats.

BACKGROUND: The Shiga toxin (Stx)-producing Escherichia coli (STEC) have become important global public health concerns. This study investigated the prevalence, antimicrobial resistance profile, and extended-spectrum beta-lactamase-producing E. coli in sheep and goat faeces. METHODS AND RESULTS: A total of 53 E. coli isolates were confirmed by PCR targeting the uidA [ß-D glucuronidase] gene. The Shiga toxin genes stx1 and stx2, as well as bfpA, vir, eaeA, lt and aafII virulence genes, were detected in this study. Of the 53 isolates confirmed to be STEC, 100% were positive for stx2 and 47.2% for stx1. Three isolates possessed a combination of stx1 + stx2 + eaeA, while four isolates harboured stx1 + stx2 + vir virulence genes. The isolates displayed phenotypic antimicrobial resistance against erythromycin (66.04%), colistin sulphate (43.4%), chloramphenicol (9.4%) and ciprofloxacin (1.9%). A total of 28.8% of the strains were phenotypically considered ESBL producers and contained the beta-lactamase blaCTX-M-9 and blaCTX-M-25 gene groups. A larger proportion of the E. coli strains (86.8%) contained the antibiotic sulphonamide resistant (sulII) gene, while 62.3%, 62.3%, 52.8%, 43.4%, 41.5%, 20.8%, 18.9%, 11.3%, 11.3%, 9.4%, 9.4% and 5.7% possessed mcr-4, floR, mcr-1, tet(A), sulI, tet(O), tet(W), parC, mcr-2, ampC 5, qnrS and ermB genes, respectively. Thirteen isolates of the ESBL-producing E. coli were considered multi-drug resistant (MDR). One Shiga toxin (stx2) and two beta-lactamase genes (blaCTX-M-9 and blaCTX-M-25 groups) were present in 16 isolates. In conclusion, the E. coli isolates from the small stock in this study contained a large array of high antibiotic resistance and virulence profiles. CONCLUSIONS: Our findings highlight the importance of sheep and goats as sources of virulence genes and MDR E. coli. From a public health and veterinary medicine perspective, the characterization of ESBL producers originating from small livestock (sheep and goats) is crucial due to their close contact with humans.

In vitro antitrypanosomal activity of synthesized nitrofurantoin-triazole hybrids against Trypanosoma species causing animal African trypanosomosis.

Animal African trypanosomosis (AAT) is a disease caused by Trypanosoma brucei brucei, T. vivax, T. evansi and T. congolense which are mainly transmitted by tsetse flies (maybe the family/genus scientific name for the tsetse flies here?). Synthetic trypanocidal drugs are used to control AAT but have reduced efficacy due to emergence of drug resistant trypanosomes. Therefore, there is a need for the continued development of new safe and effective drugs. The aim of this study was to evaluate the in vitro anti-trypanosomal activity of novel nitrofurantoin compounds against trypanosomes (Trypanosoma brucei brucei, T. evansi and T. congolense) causing AAT. This study assessed previously synthesized nineteen nitrofurantoin-triazole (NFT-TZ) hybrids against animal trypanosomes and evaluated their cytotoxicity using Madin-Darby bovine kidney cells. The n-alkyl sub-series hybrids, 8 (IC50 0.09 ± 0.02 µM; SI 686.45) and 9 (IC50 0.07 ± 0.04 µM; SI 849.31) had the highest anti-trypanosomal activity against T. b. brucei. On the contrary, the nonyl 6 (IC50 0.12 ± 0.06 µM; SI 504.57) and nitrobenzyl 18 (IC50 0.11 ± 0.03 µM; SI 211.07) displayed the highest trypanocidal activity against T. evansi. The nonyl hybrid 6 (IC50 0.02 ± 0.01 µM; SI 6328.76) was also detected alongside the undecyl 8 (IC50 0.02 ± 0.01 µM; SI 3454.36) and 3-bromobenzyl 19 (IC50 0.02 ± 0.01 µM; SI 2360.41) as the most potent hybrids against T. congolense. These hybrids had weak toxicity effects on the mammalian cells and highly selective submicromolar antiparasitic action efficacy directed towards the trypanosomes, hence they can be regarded as potential trypanocidal leads for further in vivo investigation.

Antimicrobial resistance profiles of Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae strains isolated from broiler chickens.

Globally, the spread of multidrug-resistant Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae from food to humans poses a severe threat to public health. The aim of this study was to assess the co-occurrence of colistin and ß-lactamase resistance genes in E. coli, K. pneumoniae, and P. aeruginosa strains isolated from faeces of abattoir broiler chickens. The E. coli, P. aeruginosa and K. pneumoniae isolates were successfully detected from faecal samples by polymerase chain reaction (PCR) at infection rates of 60.7%, 22.5% and 16.7% respectively. The isolates displayed the highest levels of antibiotic resistance (AR) against ampicillin (82.3%) and amoxicillin-clavulanic acid (74.2%) for E. coli, followed by cefoxitin (70.6%) for K. pneumoniae, whilst P. aeruginosa displayed 26.1% antibiotic resistance (AR) against both ampicillin and colistin sulphate. The colistin mcr-1 gene was harboured by 46.8%, 47.1% and 21.7%, E. coli, K. pneumonia and P. aeruginosa isolates respectively. Ten out of 62 (16.1%), 6/17 (35.3%), 4/23 (17.4%) isolates were phenotypically classified as ESBL E. coli, K. pneumoniae, and P. aeruginosa respectively. The ESBL-E. coli isolates respectively possessed blaCTX-M (60%), blaTEM (20%) and blaCTX-M-9 (10%) genes. The ESBL-K. pneumoniae harboured, blaCTX-M (50%), blaOXA (33%), blaCARB (17%), and blaCTX-M-9 (17%) genes respectively, whilst, P. aeruginosa isolates respectively carried blaTEM (75%), blaCTX-M (50%), blaOXA (25%) and blaCARB (25%) genes. Molecular analysis identified the blaCTX-Mß-lactamase-encoding genes collectively from E. coli, P. aeruginosa, K. pneumoniae isolates. Colistin and ß-lactamase genes were present in only 16.7%, 6.9%, and 2.9% of E. coli, K. pneumoniae, and P. aeruginosa isolates, respectively. A total of 17, 7 and 3 isolates for E. coli, K. pneumoniae and P. aeruginosa respectively carried both colistin and ß-lactamase antibiotics resistant genes. This is a public health threat that points to a challenge in the treatment of infections caused by these zoonotic bacteria. Data generated from this study will contribute to formulation of new strategies for combating spread of E. coli, K. pneumoniae, and P. aeruginosa isolates as well as prevention of their AR development.

Risk factors contributing to tick-acaricide control failure in communal areas of the Oliver Tambo district eastern cape province, South Africa.

Application of chemical acaricides in the control of ticks has led to the problem of tick-acaricide control failure. To obtain an understanding of the possible risk factors involved in this tick-acaricide control failure, this study investigated tick control practices on communal farms in the north-eastern part of the Eastern Cape Province (ECP) of South Africa. A semi-structured questionnaire designed to document specific farm attributes and acaricide usage practices was administered at 94 communal farms from the Oliver Tambo District municipality of the ECP. Data collected indicated that the main acaricide chemicals used at plunge dips of inland and coastal areas were synthetic pyrethroid formulations. Most (75%) farmers claimed not to have noticed a significant reduction in numbers of actively feeding and growing ticks on cattle after several acaricide treatments. Based on the farmers' perceptions, leading factors that could have led to tick-acaricide control failure included: weak strength of the dip solution (76%); poor structural state of dip tanks (42%); and irregular tick control (21%). The rearing of crossbreeds of local and exotic cattle breeds, perceived weak strength of the dip solution and high frequency of acaricide treatment, were statistically associated with proportions of farms reporting tick-acaricide control failure. Furthermore, approximately 50% of farms reported at least four tick control malpractices, which could have resulted in the emergence and spread of tick-acaricide control failure. Other sub-optimal tick control practices encountered included incorrect acaricide rotation, and failure to treat all cattle in a herd. This data will inform and guide the development of management strategies for tick-acaricide control failure and resistance in communal farming areas.

A 'One Health' perspective of Africa-wide distribution and prevalence of Giardia species in humans, animals and waterbodies: a systematic review and meta-analysis.

Giardiasis, caused by Giardia duodenalis, is a leading cause of diarrhoea in resource-poor countries. To gain a better insight into the epidemiology of Giardia in Africa, we undertook a robust study to comprehend the distribution and prevalence of Giardia infection in humans, animals and their dispersal in the environment. Our protocol was registered with PROSPERO (registration number CRD42022317653). Deep literature search from 5 electronic databases, namely, AJOL, Google scholar, PubMed, ScienceDirect and Springer Link was performed using relevant keywords. Meta-analysis was performed using a random-effects model and heterogeneity among studies was evaluated using Cochran's Q and the I2-statistic. More than 500 eligible studies published from 1 January 1980 until 22 March 2022 were retrieved. In humans, exactly 48 124 Giardia spp. infection cases were registered from the 494 014 stool samples examined resulting in a pooled prevalence estimate (PPE) of 8.8% using microscopy. Whereas copro-antigen tests and molecular diagnostic methods generated PPE of 14.3 and 19.5%, respectively, with HIV+ subjects and those with diarrhoeatic stool having infection rates of 5.0 and 12.3%, respectively. The PPE of Giardia spp. infection in animals using molecular methods was 15.6%, which was most prevalent in pigs (25.2%) with Nigeria registering the highest prevalence at 20.1%. The PPE of Giardia spp. contamination from waterbodies was 11.9% from a total of 7950 samples which were detected using microscopy, with Tunisia documenting the highest infection rate of 37.3%. This meta-analysis highlights the necessity of 'One Health' approach for consolidated epidemiological studies and control of giardiasis in the African continent.

"One Health" perspective on prevalence of co-existing extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae: a comprehensive systematic review and meta-analysis.

BACKGROUND: The Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) bacterial isolates that produce extended-spectrum ß-lactamases (ESBLs) contribute to global life-threatening infections. This study conducted a systematic review and meta-analysis on the global prevalence of ESBLs in co-existing E. coli and K. pneumoniae isolated from humans, animals and the environment. METHODS: The systematic review protocol was registered in the International Prospective Register of Systematic Reviews (PROSPERO) [ID no: CRD42023394360]. This study was carried out following the preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines. One hundred and twenty-six eligible studies published on co-existing antibiotic resistance in E. coli and K. pneumoniae between 1990 and 2022 were included. RESULTS: The pooled prevalence of ESBL-producing E. coli and K. pneumoniae was 33.0% and 32.7% for humans, 33.5% and 19.4% for animals, 56.9% and 24.2% for environment, 26.8% and 6.7% for animals/environment, respectively. Furthermore, the three types of resistance genes that encode ESBLs, namely blaSHVblaCTX-M,blaOXA, and blaTEM, were all detected in humans, animals and the environment. CONCLUSIONS: The concept of "One-Health" surveillance is critical to tracking the source of antimicrobial resistance and preventing its spread. The emerging state and national surveillance systems should include bacteria containing ESBLs. A well-planned, -implemented, and -researched alternative treatment for antimicrobial drug resistance needs to be formulated.

Prevalence of subclinical mastitis, its associated bacterial isolates and risk factors among cattle in Africa: a systematic review and meta-analysis.

BACKGROUND: Subclinical mastitis (SCM) is one of the most economically important diseases affecting the dairy industry. The SCM does not cause visible changes in the udder or physical changes of the milk as compared to clinical mastitis, and a clear overview of the prevalence and risk factors in the different regions of Africa is still lacking. The objective of this study was to investigate the prevalence of SCM and assess the associated risk factors and dominant bacterial pathogens among cattle in Africa. MATERIALS AND METHODS: We gathered and systematically reviewed literature concerning SCM, published in English from January 2010 through December 2020 in two databases (PubMed and Web of Science), and meta-analysis was conducted using the 'meta' and 'metafor' packages in the R statistical software. RESULTS: A total of 258 studies were retrieved and at the end of the screening, 82 full-texts were eligible for inclusion in the meta-analysis. The prevalence of SCM was reported in 11 countries in five regions of Africa, and the random-effects model showed that the weighted pooled prevalence estimate (PPE) was 48.2% (95% CI: 43.6-52.8%). Heterogeneity was high and statistically significant as I2 (proportion of observed variation) was 98.1% (95% CI: 98.0-98.3%), τ2 (true between-study variance) was 0.0433 (95% CI: 0.0322-0.0611), and the Cochran Q statistic was 4362.8 (p < 0.0001). Subgroup and meta-regression analyses showed that East Africa had significantly (p = 0.0092) the highest PPE of SCM (67.7%, 95% CI: 55.7-78.7) followed by West Africa (50.5%, 95%CI: 31.4-69.5), and the lowest was in North Africa (40.3%, 95%: 32.2-48.6). Other significant moderators for SCM were age (p < 0.0001), breed (p = 0.0002), lactation stage (p = 0.019) and parity (p = 0.0008) of cattle. Staphylococcus species (prevalence 43.7%) were the most predominant pathogens, followed by Streptococcus (18.2%) and Escherichia species (9.5%). CONCLUSION: The present study showed a high variation of SCM prevalence in various parts of Africa, although there is a need for more data in some regions. The reported prevalence is a clear sign of inappropriate management practices among cattle herds and an indicator of the threat that SCM poses to the dairy industry. The information about the predisposing factors may guide effective management and control strategies to reduce transmission of the disease.

Hematology and biochemical values in equines naturally infected with Theileria equi in Nigeria.

Equine piroplasmosis (EP) is a tick-borne disease of equines with resultant economic consequences. To date, little information is available regarding the effects of EP on the health of equids in Nigeria. Therefore, this study was carried out to ascertain the effects of natural sub-clinical infection with T. equi on the hematology and biochemical profiles of horses and donkeys. Blood and serum samples were subjected to hematology and serum biochemistry analyses, respectively. Data corresponding to PCR-positive (infected) and PCR-negative (uninfected) was computed to ascertain changes in any of the parameters. Hematological significant findings (P<0.001) in both horses and donkeys include decreased packed cell volume (PCV), red blood cell (RBC) count, hemoglobin (Hb) concentration, and white blood cell (WBC) counts in the infected group compared to the uninfected group. In horses, the PCV (%) and Hb (g/dL) values were 40.6±1.0 and 13.4±1.7 in the uninfected group compared with 30.8±0.6 and 10.1±0.3 in the infected group respectively and differs significantly (P<0.001). Similarly, in the donkeys, the PCV (%) and Hb (g/dL) values were 32.2 ± 0.5 and 10.5 ± 0.2 in the uninfected group compared with 28.5±0.7 and 9.5±0.2 in the infected group respectively with significant difference (P<0.001). Additionally, we observed a slight decrease in WBC counts (×103/µL) in the infected group in horses (9.9±0.4) and donkeys (10.2±4.3) compared with 10.5±2.0 and 13.0±0.8 from the uninfected respectively. Serum biochemical profiles revealed slight increase in the levels of total proteins principally the globulin fractions in the infected group in both equids. The findings from this study will be helpful in the diagnosis, prognosis, and treatment evaluation of equines in Nigeria.

Serosurvey for equine piroplasms in horses and donkeys from North-Western Nigeria using IFAT and ELISA.

Equine piroplasmosis is caused by apicomplexan parasites, namely, Babesia caballi and Theileria equi, which are transmitted to equids principally through ticks. To ascertain the exposure of equines to agents of equine piroplasms, we tested serum samples collected from horses (n = 272) and donkeys (n = 170) in North-Western Nigeria for the presence of antibodies against B. caballi and T. equi using IFAT and ELISA. The seroprevalence of T. equi in the horses determined using IFAT and ELISA was 48.89% and 45.96%, respectively, while for B. caballi, it was 6.3% and 0.4%, respectively. For T. equi, the seroprevalence based on IFAT and ELISA results in donkeys was 14.1% and 2.9%, respectively, while for B. caballi, the seroprevalence was 2.4% and 0.6%, respectively, for ELISA and IFAT. Mixed infection detected in the horses using IFAT and ELISA was 5.5% and 0.4%, respectively, while no mixed infection was observed in the donkeys. The seroprevalence of T. equi was significantly (P < .0001) higher than that of B. caballi in both horses and donkeys. Comparatively, the IFAT detected a greater number of piroplasm seropositive animals than ELISA, indicating a difference in their diagnostic accuracy. Findings from this study confirm the existence of equine piroplasms in both horses and donkeys in North-Western Nigeria and highlights the need for robust and effective control measures against the disease.

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.

Short- and long-term effects of orally administered azithromycin on Trypanosoma brucei brucei-infected mice.

Human African trypanosomosis (HAT) and animal African trypanosomosis (AAT) are diseases of economic importance in humans and animals that affect more than 36 African countries. The currently available trypanocidal drugs are associated with side effects, and the parasites are continually developing resistance. Thus, effective and safe drugs are needed for the treatment of HAT and AAT. This study aimed to evaluate the effects of azithromycin (AZM) on Trypanosoma brucei brucei-infected mice. Mice were randomly divided into 7 groups consisting of a vehicle control group, 5 test groups and a diminazene aceturate (DA)-treated group. Mice were treated orally for 7 and 28 days, as short-term and long-term treatments, respectively. Short-term AZM treatment cured 23% (16 of 70) of the overall treated mice whereas long-term treatment resulted in the survival of 70% of the mice in the groups that received AZM at doses of 300 and 400 mg/kg. Trypanosomes treated in vitro with 25 µg/mL of AZM were subjected to transmission electron microscopy, which revealed the presence of increased numbers of glycosomes and acidocalcisomes in comparison to the vehicle group. The current study showed the trypanocidal effect of AZM on T. b. brucei in vivo. The demonstrated efficacy increased with an increase in treatment period and an increased concentration of AZM.

Molecular characterization of a new Trypanosoma (Megatrypanum) theileri isolate supports the two main phylogenetic lineages of this species in Japanese cattle.

Trypanosoma (Megatrypanum) theileri is a cosmopolitan, usually non-pathogenic, trypanosome of cattle transmitted by blood-sucking arthropods, mainly tabanid flies. Several T. theileri strains isolated from domestic and wild ruminants via co-culturing with mammalian feeder cells or blood cells have been characterized morphologically and genetically. Here, we cultured a new trypanosome isolate from a Holstein cow in Hokkaido, Japan, and performed morphological and molecular characterization studies. The new isolate (Obihiro strain) was co-cultivated with Madin-Darby bovine kidney (MDBK) cells in GIT medium supplemented with 10% fetal bovine serum. Trypomastigotes and epimastigotes, but not intracellular parasites, were identified in the culture. Analysis of the V7-V8 region of 18S rRNA sequences showed that the Obihiro strain is positioned within the subgenus Megatrypanum. A dendrogram based on whole internal transcribed spacer rDNA sequence showed that the Obihiro strain clustered in the lineage TthII together with the Japanese isolates of T. theileri, Esashi 9, and Esashi 12, and isolates from Zambia and the USA. T. theileri of the KM strain and a T. theileri-like trypanosome isolated from deer (TSD1 strain) clustered in the lineage TthI, separate from the Obihiro strain. Based on a partial cathepsin L-like protein gene analysis, the Obihiro strain clustered with isolates of the TthIIF genotype, which includes T. theileri from Vietnam, Sri Lanka, and Brazil. Our analyses of the T. theileri Obihiro strain provide relevant insights into its genetic diversity in Japanese cattle and corroborate the host specificity of cattle and deer trypanosomes of the subgenus Megatrypanum.

Risk factors associated with occurrence of anthelmintic resistance in sheep of resource-poor farmers in Limpopo province, South Africa.

Anthelmintic treatment is the most common way of controlling nematode infections in ruminants even though several countries have reported anthelmintic resistance (AR), resulting in limitation for sustainable small ruminant production. The aim of the present study was to evaluate the knowledge of resource-poor sheep farmers in Limpopo province of South Africa on the use of anthelmintics. A questionnaire regarding helminthosis control practices was administered to small ruminant farmers in five districts of Limpopo province namely Capricorn, Sekhukhune, Waterberg, Vhembe, and Mopani. A total of 77 resource-poor farmers were interviewed between June and August of 2017 using a structured questionnaire with a combination of qualitative and quantitative open-ended questions. The interviewed farmers were divided into three groups based on their farming experience (< 5; 6-10, and ˃ 10 years of farming experience). Limited farming experience was shown as one of the risks, as farmers that owned sheep for less than 10 years could not identify the symptoms of gastrointestinal parasites infection and did not know how nematodes are transmitted to animals. However, no significant difference (p < 0.05) was found to exist between the three groups of farmers in terms of clinical signs identification and correct application of anthelmintics. About 43% of the respondents were unaware of gastrointestinal nematodes (GI) that infect sheep, could not identify the clinical symptoms of gastrointestinal nematodes infection, and only 34% knew how animals become infected. Although 67.5% of farmers mentioned that they never dose their sheep, 32.5% use anthelmintics at varying times in a year. None of the farmers weighed their sheep before dosing them instead visual appraisal of individual weight was the most common means of estimating the anthelmintic dose. The above information is an indication of risks associated with possible occurrence of anthelmintic resistance in the study areas. There is therefore, a need to give training to resource-poor farmers of small stock on proper application of anthelmintic treatment and to educate them on how to prevent development of AR. Future studies on AR should also be conducted in the province in flocks with high-treatment frequencies to establish the occurrence of AR using both in vivo and in vitro methods. The most common risk factor associated with the occurrence of AR in all the five districts of Limpopo province was found to be the use of anthelmintics without weighing the animals to determine the correct dosage.

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.

Geographic distribution of Theileria sp. (buffalo) and Theileria sp. (bougasvlei) in Cape buffalo (Syncerus caffer) in southern Africa: implications for speciation.

Strict control measures apply to movement of buffalo in South Africa including testing for Theileria parva, the causative agent of Corridor disease in cattle. The official test is a real-time hybridization PCR assay that amplifies the 18S rRNA V4 hyper-variable region of T. parva, T. sp. (buffalo) and T. sp. (bougasvlei). Mixed infections with the latter organisms affect diagnostic sensitivity due to PCR suppression. While the incidence of mixed infections in the Corridor disease endemic region of South Africa is significant, little information is available on the specific distribution and prevalence of T. sp. (buffalo) and T. sp. (bougasvlei). Specific real-time PCR assays were developed and a total of 1211 samples known to harbour these parasites were screened. Both parasites are widely distributed in southern Africa and the incidence of mixed infections with T. parva within the endemic region is similar (∼25-50%). However, a significant discrepancy exists in regard to mixed infections of T. sp. (buffalo) and T. sp. (bougasvlei) (∼10%). Evidence for speciation between T. sp. (buffalo) and T. sp. (bougasvlei) is supported by phylogenetic analysis of the COI gene, and their designation as different species. This suggests mutual exclusion of parasites and the possibility of hybrid sterility in cases of mixed infections.

Parasitic infection among HIV/AIDS patients at Bela-Bela clinic, Limpopo province, South Africa with special reference to Cryptosporidium.

Intestinal parasitic organisms are common pathogens among HIV patients worldwide and have been known to cause severe and life-threatening diarrhea in such subjects. In the present study, the prevalence of Cryptosporidium spp and other intestinal parasites in stool samples from 151 HIV/AIDS patients attending a HIV treatment center in South Africa was determined using' standard parasitological methods, as well as molecular methods including PCR and quantitative PCR for confirmation of Cryptosporidium spp. In addition, the loop-mediated isothermal amplification (LAMP) method was evaluated for detection of Cryptosporidium spp in 24 stool samples. Standard parasitological methods indicated that Cryptospo- ridium spp (26.5%), Entamoeba spp (26.5%) and Giardia lamblia (13%) were the most common protozoan parasites, while Ascaris lumbricoides (8%), Schistosoma mansoni (6%) and Trichuris trichiura (4.6%) were the most commonly found helminths. PCR, quantitative PCR and LAMP methods identified Cryptosporidium spp in 28% (30/106), 35% (53/151) and 58% (14/24) of the stool samples, respectively. Multiple infections (34%) were commonly found in the study population. Females above 45 years had the highest Cryptosporidium prevalence (58%). Prevention measures must be implemented in order to curb the negative impact of Cryptosporidium-causing diarrhea among HIV/AIDS patients in this region as well as other parasitic infections identified in this study.

Accuracy of molecular diagnostic assays for detection of Mycobacterium bovis: A systematic review and meta-analysis.

Bovine tuberculosis (bovine TB) is a chronic wasting disease of cattle caused primarily by Mycobacterium bovis. Controlling bovine TB requires highly sensitive, specific, quick, and reliable diagnostic methods. This systematic review and meta-analysis evaluated molecular diagnostic tests for M. bovis detection to inform the selection of the most viable assay. On a per-test basis, loop-mediated isothermal amplification (LAMP) showed the highest overall sensitivity of 99.0% [95% CI: 86.2%-99.9%] and specificity of 99.8% [95% CI: 96.2%-100.00%]. Quantitative real-time polymerase chain reaction (qPCR) outperformed conventional PCR and nested PCR (nPCR) with a diagnostic specificity of 96.6% [95% CI: 88.9%-99.0%], while the diagnostic sensitivity of 70.8% [95% CI: 58.6-80.5%] was comparable to that of nPCR at 71.4% [95% CI: 60.7-80.2%]. Test sensitivity was higher with the input of milk samples (90.9% [95% CI: 56.0%-98.7%]), while specificity improved with tests based on major M. bovis antigens (97.8% [95% CI: 92.3%-99.4%]), the IS6110 insertion sequence (95.4% [95% CI: 87.6%-98.4%]), and the RD4 gene (90.7% [95% CI: 52.2%-98.9%]). The design of the currently available molecular diagnostic assays, while mostly based on nonspecific gene targets, prevents them from being accurate enough to diagnose M. bovis infections in cattle, despite their promise. Future assay development should focus on the RD4 region since it is the only target identified by genome sequence data as being distinctive for detecting M. bovis. The availability of a sufficiently accurate diagnostic test combined with the routine screening of milk samples can decrease the risk of zoonotic transmissions of M. bovis.

Comprehensive whole genome analysis of Staphylococcus aureus isolates from dairy cows with subclinical mastitis.

Staphylococcus species are the primary cause of mastitis in dairy cows across the world. Staphylococcus aureus has recently become a pathogen that is zoonotic and multidrug resistant. This study aimed to sequence whole genomes of 38 S. aureus isolates from 55 subclinical mastitis dairy cows of 7 small-scale farmers in the Free State Province, South Africa and document and their antimicrobial and virulence genes. The 38 isolates were grouped by the in silico multi-locus sequencing types (MLST) into seven sequence types (STs), that is (ST 97, 352, 152, 243) and three new STs (ST8495, ST8500, and ST8501). Thirty-three S. aureus isolates were divided into 7 core single-nucleotide polymorphism (SNP) clusters. Among the 9 distinct spa-types that were detected, Spa-types t2883 accounted for the majority of isolates at 12 (31.57%), followed by t416 with 11 (28.94%) and t2844 with 5 (13.15%). The data also revealed the identification of four (4) plasmids, with Rep_N (rep20) accounting for the majority of isolates with 17 (44.73%), followed by Inc18 (repUS5) with 2 (5.26%). These isolates included 11 distinct antimicrobial resistance genes and 23 genes linked to bacterial virulence. Surprisingly, no methicillin resistance associated genes were detected in these isolates. Genome data of the current study will contribute to understanding epidemiology S. aureus genotypes and ultimately aid in developing treatment and control plans to stop the spread of mastitis in the Free State province and South Africa as a whole.

Coxiella burnetii infections from animals and ticks in South Africa: a systematic review.

Coxiella burnetii is a zoonotic intracellular bacterium that is widely distributed and affects domestic animals, wildlife, humans and non-mammalian species. This systematic review was aimed at synthesizing research findings on C. burnetii in both domestic and wild animals of South Africa. The systematic review protocol was registered with Open Society Foundations of systematic reviews ( https://doi.org/10.17605/OSF.IO/8WS ). PRISMA guidelines were followed to collect and evaluate relevant scientific articles published on C. burnetii infecting domestic and wild animals in South Africa. Published articles were sourced from five electronic databases, namely, Google Scholar, PubMed and ScienceDirect, EBSCO and Scopus. Results showed 11 eligible studies involving four domestic animals, three wild animals and one ectoparasite species from seven provinces across South Africa. The occurrence of C. burnetii infection was high in Ceratotherium simum (white rhinoceros) (53.9%), medium in sheep (29.0%) and low in pigs (0.9%). Limpopo province (26%) had the most recorded infections followed by KwaZulu-Natal (19%) and Free State (3%) had the least reported occurrence of C. burnetii. The current study discovered that there is scarcity of published research on prevalence and distribution of C. burnetii infecting domestic and wild animals in South Africa, and this is of concern as this bacterium is an important zoonotic pathogen of "One Health" importance.