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Background: Canine babesiosis and ehrlichiosis are tick-borne infections of great significance in South Africa. Theileriosis in dogs in South Africa is still poorly understood. Co-infection with multiple tick-borne diseases has been documented and is perceived as a common occurrence in South Africa., Objectives: The main objective of this study was to determine the prevalence of co-infections with Ehrlichia canis or Theileria equi in dogs with babesiosis in the Eastern Cape province. There is a lack of data on canine tick-borne disease distribution in this region. Possible associations of population characteristics and haematological and biochemistry measures with a co-infection of E. canis or T. equi in these dogs were also investigated., Method: The study population included 150 dogs naturally infected with babesiosis that presented to the Mdantsane State Veterinary Clinic between January 2021 and November 2021. Quantitative polymerase chain reaction was used to confirm the Babesia spp. that the dogs were infected with and to identify co-infections. Association with co-infection for the following parameters were evaluated: sex, breed, age, duration of illness, leukocyte count, band neutrophil count, monocyte count, platelet count, ARC, and serum globulin concentration. Positive and negative predictive values of monocytosis, leukopenia, band neutrophilia, thrombocytopenia, and non-regenerative absolute reticulocyte count for co-infection were also calculated., Results: Babesia rossi was identified in 149/150 samples and B. vogeli in only 1/150 samples. A co-infection prevalence of 2.0% (3/149; 95% CI: 0.4-5.7) with B. rossi and E. canis was found. No other co-infections were reported. No investigated variables showed significant associations with co-infections. Monocytosis, in particular, was not associated with co-infection., Conclusion: Co-infection with other tick-borne diseases in dogs with babesiosis is uncommon in the Eastern Cape province. These findings raise the possibility that B. rossi may have a protective effect against other tick-borne diseases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper: Yolandi Rautenbach reports financial support was provided by Health and Welfare Sector Education and Training Authority (HWSETA). If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

We used whole genome sequencing (WGS) as an epidemiologic surveillance tool to elucidate the transmission dynamics of Shiga toxin-producing Escherichia coli (STEC) strains along the beef production chain in South Africa. Isolates were obtained from a cattle farm, abattoirs and retail outlets. Isolates were analysed using WGS on a MiSeq platform (Illumina, San Diego, CA, USA) and phylogenetic analysis was carried out. Of the 85 isolates, 39% (33) carried the stx gene and 61% (52) had lost the stx gene. The prevalence of stx subtypes was as follows; stx 1 a 55% (18/33), stx 1 b 52% (17/33), stx 2 a 55% (18/33), stx 2 b 27% (9/33), stx 2 dB 30% (10/33) and stx 2 d 1 A 15% (5/33). Thirty-five different serogenotypes were detected, of which 65% (56) were flagellar H-antigens and 34% (29) were both O-antigens and flagellar H-antigens. We identified 50 different sequence types (STs), and only nine of the isolates were assigned to three different clonal complexes. Core genome phylogenetic analysis revealed genetic relatedness, and isolates clustered mainly according to their STs and serogenotypes regardless of stx subtypes. This study provides evidence of horizontal transmission and recirculation of STEC strains in Gauteng province and demonstrates that every stage of the beef production chain plays a significant role in STEC entry into the food chain.

Bovine neosporosis is a widespread parasitic disease associated with significant economic losses. Its effects on the reproductive performance of cows have resulted in losses that run into the hundreds of millions of US dollars in dairy industries in various countries (Reichel et al., Int J Parasitol 43:133-142, 2013). Due to outdated and scant information on the occurrence of Neospora caninum infection in South Africa, the study aimed to determine the seroprevalence and risk factors associated with infection in dairy cattle in South Africa. A total of 1401 blood samples were randomly collected from cattle on 48 dairy farms in seven of the nine provinces in South Africa. A close-ended questionnaire was used in a cross-sectional study to obtain farm-level and animal-level data. Serological testing was done using a commercial IDvet Screen® Neospora caninum Indirect ELISA. An overall seroprevalence, adjusted for test sensitivity and specificity, of 2.3% (95% CI, 1.3-4.1) was detected and 48% (23/48) of sampled farms had at least one animal testing positive. The highest seroprevalence of N. caninum was in the KwaZulu-Natal province with 7.5% (95% CI, 3.8-14.3), and the lowest in Western Cape with 0.1% (95% CI, 0-1.2). The highest within-farm seroprevalence of 25% was detected on a farm in the North West Province. In a multivariable logistic regression model, the odds of N. caninum seropositivity were higher in Holstein-Friesian cattle when compared to other breeds. Good hygiene was identified as a protective factor. Cattle left out on pasture had increased odds of testing positive for N. caninum compared to those that were penned. The odds of testing seropositive for N. caninum was higher on farms that practised segregation of cattle into different age groups. The purchase of replacement animals was a significant risk factor, as open herds had increased odds of N. caninum seropositivity. Cattle on farms that did not have a specific calving location were more likely to be seropositive. This is the first such study in South Africa and shows that N. caninum is widely distributed in the country at a low seroprevalence, but it may be a cause of concern on certain farms., (© 2024. The Author(s).)

Background: Rift Valley fever virus (RVFV) is a zoonotic mosquito-borne virus with serious implications for livestock health, human health, and the economy in Africa, and is suspected to be endemic in north-eastern KwaZulu-Natal (KZN), South Africa. The vectors of RVFV in this area are poorly known, although several species, such as Aedes (Neomelaniconion) mcintoshi, Aedes (Neomelaniconion) circumluteolus, Aedes (Aedimorphus) durbanensis, and Culex (Lasioconops) poicilipes may be involved. The aim of the study was to determine the vertebrate blood meal sources of potential RVFV mosquito vectors in north-eastern KZN and to characterize the host-biting network., Methods: Blood-fed mosquitoes were collected monthly from November 2019 to February 2023 using a backpack aspirator, CO 2 -baited Centers for Disease Control and Prevention (CDC) miniature light traps and tent traps, in the vicinity of water bodies and livestock farming households. The mosquitoes were morphologically identified. DNA was extracted from individual mosquitoes and used as templates to amplify the vertebrate cytochrome c oxidase I (COI) and cytochrome b (cytb) genes using conventional polymerase chain reaction (PCR). Amplicons were sequenced and queried in GenBank and the Barcode of Life Data systems to identify the vertebrate blood meal sources and confirm mosquito identifications. All mosquitoes were screened for RVFV using real time reverse transcription (RT)-PCR., Results: We identified the mammalian (88.8%) and avian (11.3%) blood meal sources from 409 blood-fed mosquitoes. Aedes circumluteolus (n = 128) made up the largest proportion of collected mosquitoes. Cattle (n = 195) and nyala (n = 61) were the most frequent domestic and wild hosts, respectively. Bipartite network analysis showed that the rural network consisted of more host-biting interactions than the reserve network. All mosquitoes tested negative for RVFV., Conclusions: Several mosquito species, including Ae. circumluteolus, and vertebrate host species, including cattle and nyala, could play a central role in RVFV transmission. Future research in this region should focus on these species to better understand RVFV amplification., (© 2024. The Author(s).)

Background: The stress response after acute trauma in humans and animals has been well-recognised in the literature. However, data on temporal changes in endocrine parameters after acute trauma in previously healthy dogs are scant., Objective: To longitudinally track endocrine variables and lactate concentrations in dogs with canine bite wounds., Method: Prospective study involving 20 dogs hospitalised after being bitten by another dog. Serum cortisol, thyroxine (total T4), thyrotropin (TSH), and lactate concentrations were measured on admission and every 8 hours after the recorded bite incident, for a 72-hour period., Results: Median cortisol concentration was markedly elevated on admission (314.6 nmol/L; IQR 229.3-369.6) but returned to within the reference interval by 16 hours post-bite (99.5 nmol/L; IQR 48.7-225.4) and reached a nadir at 48 hours post-bite (38.5 nmol/L; IQR 32.1-115.9). Median total T4 concentration was within the reference interval on admission (20.6 nmol/L; IQR 12.7-27.9) but decreased below the reference interval by 8 hours (11.0 nmol/L; IQR 5.0-14.1) and reached a nadir 16 hours postbite (7.0 nmol/L; IQR 2.9-19.7), before gradually increasing after 64 hours to reach the reference range by 72 hours. The median TSH concentrations remained within the reference interval throughout the study period. Median lactate concentration was mildly elevated on admission (3.0 mmol/L; IQR 2.0-3.9)., Conclusion: This study produced novel data on the temporal relationships of the hypothalamic-pituitary-adrenal and hypothalamicpituitary- thyroidal axes alterations after an acute traumatic insult in dogs.

Background: Q fever and toxoplasmosis are economically important zoonoses as they cause considerable losses in livestock (cattle, sheep and goats) and wildlife (antelopes, giraffes, lions, and cheetahs) through reproductive disorders such as abortions and stillbirths. Q fever and toxoplasmosis testing in South Africa is conducted by the Agricultural Research Council-Onderstepoort Veterinary Research (ARC-OVR). However, both zoonoses are understudied and not monitored in South Africa as they are not considered controlled or notifiable diseases in the Animal Disease Act 35 of 1984. A retrospective study was conducted on Q fever (2007-2009) and toxoplasmosis (2007-2017) using diagnostic laboratory data at the ARC-OVR. Also, we report on sporadic abortion and stillbirth cases in livestock from diagnostic tissue samples submitted for Coxiella burnetii polymerase chain reaction (PCR) detection at the ARC-OVR., Results: During 2007 to 2009, 766 animal samples were tested for C. burnetii antibodies and seropositivity was 0.9% (95%CI: 0.3-1.7) with sheep (1.9%; 95%CI: 0.6-4.4) having the highest seropositivity followed by cattle (0.7%; 95%CI: 0.09-2.6), while all goats (0.0%; 95%CI: 0.0-4.2) and wildlife (0.0%; 95%CI: 0.0-2.5) tested were negative. From 2007 to 2017, 567 sera were tested for T. gondii antibodies; overall seropositivity was 12.2% (95%CI: 9.6-15). Wildlife had highest seropositivity to T. gondii antibodies (13.9%; 95%CI: 9.0-19.7) followed by goats (12.9%; 95%CI: 9.2-17.4) and sheep (12.3%; 95%CI: 5.1-23.8) while seropositivity in cattle was 2.4% (95%CI: 0.06-12.9). Of 11 animals tested by C. burnetii PCR detection (2021-2022), 10 (91.0%) were positive. The amplicon sequences showed similarity to Coxiella burnetii strain 54T1 transposase gene partial coding sequence., Conclusions: We have confirmed the occurrence of the causative agents of Q fever and toxoplasmosis in livestock and wildlife in South Africa, with data limitations. These zoonoses remain of importance with limited information about them in South Africa. This study provides baseline information for future studies on Q fever and toxoplasmosis in South African livestock and wildlife, as well other African countries. Due to limited data collection experienced in this study, it is recommended that improvements in data collection samples tested should include associated factors such as sex, age, and breed of the animals., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Rift Valley fever (RVF) is a re-emerging zoonotic disease of domestic ruminants and humans. While neighbouring countries have reported outbreaks of RVF, Ghana has not yet identified any cases. The aim of this study was to determine whether RVF virus (RVFV) was circulating in livestock and herders in the southern part of Ghana, to estimate its seroprevalence, and to identify associated risk factors. The study surveyed 165 livestock farms randomly selected from two districts in southern Ghana. Serum samples of 253 goats, 246 sheep, 220 cattle, and 157 herdsmen were tested to detect IgG and IgM antibodies against RVFV. The overall seroprevalence of anti-RVF antibodies in livestock was 13.1% and 30.9% of farms had RVFV seropositive animals. The species-specific prevalence was 24.1% in cattle, 8.5% in sheep, and 7.9% in goats. A RVFV IgG seroprevalence of 17.8% was found among the ruminant herders, with 8.3% of all herders being IgM positive. RVFV was shown, for the first time, to have been circulating in southern Ghana, with evidence of a recent outbreak in Kwahu East; however, it was clinically undetected despite significant recent human exposure. A One Health approach is recommended to better understand RVF epidemiology and socio-economic impact in Ghana.

Infection with Coxiella burnetii causes significant economic impact and poses zoonotic risk to people exposed to livestock, yet few studies in South Africa have assessed seroprevalence of C. burnetii infection and no information is available for goats. Very little information is available regarding risk factors and outcomes of C. burnetii infection in peri-urban farming areas where widespread mixing of ruminants occurs. This study estimated the seroprevalence of C. burnetii infection among communally farmed goats in an area adjacent to the densely populated Gauteng province. Sera were collected from 216 goats in 39 herds, and questionnaires were completed to establish management practices as potential risk factors. C. burnetii antibody testing was done by ELISA. Thirty two out of 216 goats tested positive for C. burnetii antibodies and the overall seroprevalence, adjusted for sampling weights and clustering, was 18.4% (95% confidence interval [CI]: 12.2% - 23.5%). The intraclass correlation coefficient was 0.06, indicating low-to-moderate clustering. Multiple logistic regression showed age was significantly associated with seropositivity, with higher seroprevalence in animals ≥ 19 months old (26%) than animals ≤ 6 months old (6%) (odds ratio [OR]: 6.6; p = 0.010). We concluded C. burnetii infection is common in goats in Moretele and a potential cause of abortion in goats and poses the potential zoonotic disease risk.Contribution: Despite the threats posed on animal health and productivity, scant information is published on C. burnetii in South Africa. This research established preliminary estimates of C. burnetii seroprevalence. The research is original from a South African perspective, relevant to Africa and focused on infectious disease in livestock.

An experimental infection using Babesia (B.) rossi was performed in healthy male Beagle dogs to assess the changes in endocrine variables during disease. Two dogs were infected with a low dose (LD) of parasite inoculum (10 4 parasites) and three dogs were infected with a high dose (HD) (10 8 parasites). Basal serum cortisol, thyroxine (T4), and thyrotropin (TSH) concentrations were measured every second day. Samples were analyzed using a solid- phase, competitive chemiluminescent enzyme immunoassay (Immulyte® 2000, Siemens). Variables were compared between groups and timepoints using linear mixed models. In both groups, the median cortisol concentration increased, whilst the median T4 concentration decreased after infection, with a return towards baseline concentration post treatment. The highest cortisol and the lowest T4 concentrations were reached at 96 h and 108 h post infection, respectively, in the HD group and slightly later at 108 and 144 h post-infection, respectively, in the LD group. A higher cortisol concentration with a more rapid increase, and a lower T4 concentration with a more rapid decline, were associated with disease severity and a higher dose of parasite inoculum. The TSH concentration remained within the reference interval throughout the study period. This study illustrated the temporal changes in endocrine parameters during experimental B. rossi infection and demonstrated that cortisol and T4 tracked the severity of disease, albeit in opposite directions., Competing Interests: Declaration of Competing Interest None., (Copyright © 2022. Published by Elsevier GmbH.)

Abortions in domestic ruminants cause significant economic losses to farmers. Determining the cause of an abortion is important for control efforts, but it can be challenging. All available diagnostic methods in the bacteriology laboratory should be employed in every case due to the many limiting factors (autolysis, lack of history, range of samples) that complicate the investigation process. The purpose of this study was to determine whether the recovery of diagnostically significant isolates from domestic ruminant abortion cases could be increased through the use of a combination of the existing aerobic culture and Brucella selective method with methods that are commonly recommended in the literature reporting abortion investigations. These methods are examination of wet preparations and impression smears stained by the modified Ziehl-Neelsen method, anaerobic, microaerophilic, Leptospira, Mycoplasma and fungal culture. Samples of placenta and aborted foetuses from 135 routine clinical abortion cases of cattle (n = 88), sheep (n = 25) and goats (n = 22) were analysed by the new combination of methods. In 46 cases, bacteria were identified as aetiological agents and in one case a fungus. Isolation of Brucella species increased to 7.4% over two years compared with the previous 10 years (7.3%), as well as Campylobacter jejuni (n = 2) and Rhizopus species (n = 1). Salmonella species (5.9%) and Trueperella pyogenes (4.4%) were also isolated more often. In conclusion, the approach was effective in removing test selection bias in the bacteriology laboratory. The importance of performing an in-depth study on the products of abortion by means of an extensive, combination of conventional culture methods was emphasised by increased isolation of Brucella abortus and isolation of C. jejuni. The combination of methods that yielded the most clinically relevant isolates was aerobic, microaerophilic, Brucella and fungal cultures.

This study explores the impact of four-day school weeks on early elementary achievement. Using covariate adjusted regression analyses and data on all students who entered kindergarten in Oregon, USA between 2014 and 2016, we examine differences in 3rd grade math and English Language Arts test scores (i.e., achievement) for students enrolled in a four-day school week versus a five-day school week at kindergarten entry . On average, we find minimal differences between 3 rd grade test scores of four-day and five-day students, but there are notable differential effects across the spectrum of these students' kindergarten readiness scores and educational program participation. We find that above median performers on kindergarten assessments, White students, general education students, and gifted students - student groups that make up more than half our sample - are the most negatively impacted by the four-day school week during the early elementary period. We generally find no statistically significant evidence of detrimental four-day school week achievement impacts for students who were below median performers on kindergarten assessments, minority students, economically disadvantaged students, special education participants, and English as a second language students.

The presence of zoonotic diseases adversely affects livestock production and farmers' livelihood in communal areas. A lack of awareness about zoonotic diseases among rural farmers results in economic losses and health risks. The far north-eastern corner of KwaZulu-Natal (KZN) province, South Africa, is home to large numbers of communal livestock farmers who live adjacent to wildlife reserves and international borders. There have been reports of zoonotic and trade-sensitive diseases in the area, but farmers' knowledge, attitudes, and practices (KAP) in this regard are poorly understood. This study investigated the KAPs of communal livestock farmers in far northern KZN regarding livestock and zoonotic diseases found in the livestock-wildlife interface and determined the constraints and challenges faced by communal livestock farmers. A cross-sectional questionnaire survey was conducted among 504 livestock farmers at 45 dip tanks between August and November 2020, using a closed-ended questionnaire. Although the overall level of knowledge regarding animal disease transmission was fairly good (score: 53.2%), 25.4% and 21.4% of farmers had moderate and poor knowledge, respectively, about zoonotic disease transmission and prevention. Over 40% of the farmers were not aware of the zoonotic nature of wildlife and livestock diseases. Older farmers, despite their lower level of education, were more knowledgeable on animal diseases and had better practices in regard to zoonotic disease prevention and management compared to younger ones. The majority of farmers cited the lack of water, insufficient grazing land, stock theft, the restriction of animal movement, and animal diseases as the most significant challenges they faced regarding animal production. The results indicate the need for extension programs that target educating livestock farmers to improve their knowledge of these diseases.

Background: Bovine tuberculosis (bTB) is a zoonotic disease with great economic impact estimated at billions of dollars annually worldwide. Meat inspection represents a long-standing form of disease surveillance that serves both food safety and animal health. The objective of this study was to determine the prevalence of bTB in livestock at abattoirs using a cell-mediated immune (CMI) assay, the gamma interferon (IFN-γ) assay. This cross-sectional study was conducted at selected abattoirs (low-throughput, high-throughput and rural/informal) in Gauteng province, where animals were also subjected to routine meat inspection., Results: A total of 410 fresh blood samples were collected from slaughter livestock (369 cattle and 41 sheep) from 15 abattoirs, and analysed using Bovigam ® test kit with bovine, avian and Fortuitum purified protein derivatives (PPD) as blood stimulating antigens. The estimated prevalence of bTB in cattle was 4.4% (95% CI: 2.4%-7.3%). The prevalence of bTB in cattle varied between abattoirs (p = .005), ranging from 0% to 23%; however, there were no significant differences among genders, breeds, municipality, districts, origins of animals (feedlot, auction or farm) or throughput of abattoirs. The prevalence of avian reactors was 6.0% (95% CI: 3.6%-9.2%) in cattle, varying between abattoirs (p = .004) and ranging from 0% to 20.7%. None of the sheep with valid test results was positive for bTB and none was avian reactors (95% CI: 0%-15%)., Conclusion: The detection of bTB reactor cattle in our study clearly shows the limitation of disease surveillance using a meat inspection approach, as all the 410 slaughter animals sampled had passed visual abattoir inspection and been classified as bTB-free. Our findings therefore emphasize the risk of zoonotic transmission of bTB to abattoir workers and potential food safety hazard to consumers. Furthermore, our study highlights the potential for the use of the IFN-γ assay to reduce this risk., (© 2021 Agricultural Research Council. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Background: Crimean-Congo haemorrhagic fever (CCHF) is a priority emerging pathogen for which a licensed vaccine is not yet available. We aim to assess the feasibility of conducting phase III vaccine efficacy trials and the role of varying transmission dynamics., Methods: We calibrate models of CCHF virus (CCHFV) transmission among livestock and spillover to humans in endemic areas in Afghanistan, Turkey and South Africa. We propose an individual randomised controlled trial targeted to high-risk population, and use the calibrated models to simulate trial cohorts to estimate the minimum necessary number of cases (trial endpoints) to analyse a vaccine with a minimum efficacy of 60%, under different conditions of sample size and follow-up time in the three selected settings., Results: A mean follow-up of 160,000 person-month (75,000-550,000) would be necessary to accrue the required 150 trial endpoints for a target vaccine efficacy of 60 % and clinically defined endpoint, in a setting like Herat, Afghanistan. For Turkey, the same would be achieved with a mean follow-up of 175,000 person-month (50,000-350,000). The results suggest that for South Africa the low endemic transmission levels will not permit achieving the necessary conditions for conducting this trial within a realistic follow-up time. In the scenario of CCHFV vaccine trial designed to capture infection as opposed to clinical case as a trial endpoint, the required person-months is reduced by 70 % to 80 % in Afghanistan and Turkey, and in South Africa, a trial becomes feasible for a large number of person-months of follow-up (>600,000). Increased expected vaccine efficacy > 60 % will reduce the required number of trial endpoints and thus the sample size and follow-time in phase III trials., Conclusions: Underlying endemic transmission levels will play a central role in defining the feasibility of phase III vaccine efficacy trials. Endemic settings in Afghanistan and Turkey offer conditions under which such studies could feasibly be conducted., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Biosecurity measures have been introduced to limit economic losses and zoonotic exposures to humans by preventing and controlling animal diseases. However, they are implemented on individual farms with varying frequency. The goal of this study was to evaluate which biosecurity measures were used by farmers to prevent infectious diseases in ruminant livestock and to identify factors that influenced these decisions. We conducted a survey in 264 ruminant livestock farmers in a 40,000 km 2 area in the Free State and Northern Cape provinces of South Africa. We used descriptive statistics, to characterize biosecurity measures and farm attributes, then multivariable binomial regression to assess the strength of the association between the attributes and the implementation of biosecurity measures including property fencing, separate equipment use on different species, separate rearing of species, isolation of sick animals, isolation of pregnant animals, quarantine of new animals, animal transport cleaning, vaccination, tick control and insect control. Ninety-nine percent of farmers reported using at least one of the 10 biosecurity measures investigated (median [M]: 6; range: 0-10). The most frequently used biosecurity measures were tick control (81%, 214 out of 264), vaccination (80%, 211 out of 264) and isolation of sick animals (72%, 190 out of 264). More biosecurity measures were used on farms with 65-282 animals (M: 6; odds ratio [OR]: 1.52) or farms with 283-12,030 animals (M: 7; OR: 1.87) than on farms with fewer than 65 animals (M: 4). Furthermore, farmers who kept two animal species (M: 7; OR: 1.41) or three or more species (M: 7) used more biosecurity measures than single-species operations (M: 4). Farmers with privately owned land used more biosecurity measures (M: 6; OR: 1.51) than those grazing their animals on communal land (M: 3.5). Farms that reported previous Rift Valley fever (RVF) outbreaks used more biosecurity measures (M: 7; OR: 1.25) compared with farms without RVF reports (M: 6) and those that purchased animals in the 12 months prior to the survey (M: 7; OR: 1.19) compared with those that did not (M: 6). When introducing new animals into their herds (n = 122), most farmers used fewer biosecurity measures than they did for their existing herd: 34% (41 out of 122) used multiple biosecurity measures like those of vaccination, tick control, quarantine or antibiotic use, whereas 36% (44 out of 122) used only one and 30% (37 out of 122) used none. Certain farm features, primarily those related to size and commercialization, were associated with more frequent use of biosecurity measures. Given the variation in the application of biosecurity measures, more awareness and technical assistance are needed to support the implementation of a biosecurity management plan appropriate for the type of farm operation and available resources., (© 2022 The Authors. Transboundary and Emerging Diseases published by Wiley-VCH GmbH.)

In South Africa, there is a shortage of epidemiologic data on Shiga toxin-producing Escherichia coli (STEC) in the beef production chain. This study was conducted to characterise STEC isolates originating from three studies conducted in a cattle feedlot, beef abattoirs and retail outlets in Gauteng province, South Africa. Polymerase chain reaction was used to detect virulence genes, the Epsilometer test to assess antimicrobial susceptibility, pulsed-field gel electrophoresis (PFGE) to investigate genetic relatedness of isolates, and conventional serotyping for phenotypic identification. Amongst the 86 STEC isolates, the eaeA gene was detected in 20 (23%), and 26 different serogroups were identified, including the clinically important O8, O174, O2, 020 and O117. The majority of the isolates (95%; 82/86) exhibited resistance to one or more antimicrobial agents, and 30 of the isolates (35%) exhibited multi-drug resistance (MDR), being resistant to at least three antimicrobial classes. The PFGE patterns showed a highly diverse but related STEC population, with 45 distinct patterns and evidence of horizontal transmission along the beef production chain. This is significant because it demonstrates continual environmental contamination and risk of contamination along the beef production chain and the food chain. To our knowledge, this is the first study that provides evidence of horizontal transmission of STEC along the beef production chain in South Africa. This epidemiological information could facilitate the development of a proactive strategy for reducing potential foodborne outbreaks and transmission of antimicrobial resistant pathogens in the food chain., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Rabies is a global viral zoonosis endemic to South Africa, resulting in fatal encephalitis in warm-blooded animals, including humans. The loss of human lives and economic losses in rural areas through loss of livestock are substantial. A review was conducted of all confirmed animal rabies cases in South Africa from 1993 to 2019, with a total of 11 701 cases identified to species level to assess the role that wildlife plays in the epidemiology of rabies. A spatio-temporal cluster analysis using a discrete Poisson space-time probability model, accounting for underlying estimated dog and livestock densities, identified 13 significant clusters (p < .05). These included four long-term clusters lasting more than 8 years in duration and seven short-term clusters lasting less than 2 years, with the remaining two clusters being of intermediate length. Outside of these endemic clusters, wildlife outbreaks in the remainder of South Africa were often less than one and a half years in duration most likely due to the rapid decline of wildlife vectors, especially jackals associated with rabies infection. Domestic dogs accounted for 59.8% of cases, with domestic cats (3.2%), livestock (21.1%) and wildlife (15.8%) making up the remainder of the cases. Yellow mongoose (Cynictis penicillata) was the most frequently affected wildlife species, followed by bat-eared fox (Otocyon megalotis), black-backed jackal (Canis mesomelas), meerkat (Suricata suricatta) and aardwolf (Proteles cristatus). Rabies in wildlife species followed different spatial distributions: black-backed jackal cases were more common in the north-western parts of South Africa, yellow mongoose cases more frequent in central South Africa, and bat-eared fox and aardwolf cases were more frequent in southern and western South Africa. Clusters often spanned several provinces, showing the importance of coordinated rabies control campaigns across administrative boundaries, and high-risk areas were highlighted for rabies in South Africa., (© 2021 Wiley-VCH GmbH.)

Rabies is a zoonotic disease that remains endemic in large parts of southern Africa because of its persistence in wildlife and domestic dog vectors. The black-backed jackals (Canis mesomelas) is primarily the wildlife vector responsible for rabies outbreaks in northern parts of South Africa. Two trials were carried out to investigate antibody responses to the oral rabies vaccine Raboral V-RG® in black-backed jackals under captive and free-ranging conditions. In captive jackals 10/12 (83%; 95% confidence interval [CI]: 52% - 98%), seroconverted after single oral vaccination. Nine captive jackals had protective antibody titres ( 0.5 IU/mL) at 4 weeks (median: 2.1 IU/mL; inter quartile range [IQR]: 0.6-5.7) and 10 jackals had at 12 weeks (median: 3.5 IU/mL; IQR: 1.5-8.3) and three maintained antibody titres for up to 48 weeks (median: 3.4 IU/mL; IQR: 2.0-6.3). Four sites were baited with Raboral V-RG® vaccine for wild jackals, using fishmeal polymer and chicken heads. Baits were distributed by hand or from vehicle at three sites in north-eastern South Africa, with an average baiting density of 4.4 baits/km2 and at one site in central South Africa, at 0.12 baits/km2. This resulted in protective antibody titres in 3/11 jackals (27%; 95% Cl: 6-61) trapped between 3 and 12 months after baiting in north-eastern South Africa, compared with 4/7 jackals (57%; 95% Cl: 18-90) trapped after 3-18 months in central South Africa. This study shows the potential utility of oral rabies vaccination for the control of wildlife-associated rabies in north-eastern and central South Africa, but extensive studies with wider distribution of bait are needed to assess its potential impact on rabies control in wild jackals.

Rift Valley fever virus (RVFV) is a mosquito-borne, zoonotic phlebovirus-causing disease in domestic ruminants and humans in Africa, the Arabian Peninsula and some Indian Ocean islands. Outbreaks, characterized by abortion storms and a high morbidity rate in newborn animals, occur after heavy and prolonged rainfalls favouring the breeding of mosquitoes. However, the identity of the important mosquito vectors of RVFV is poorly known in most areas. Mosquitoes collected in the Ndumo area of tropical north-eastern KwaZulu-Natal (KZN), South Africa, were tested for RVFV nucleic acid using RT-PCR. The virus was detected in a single pool of unfed Aedes ( Aedimorphus ) durbanensis , indicating that this seasonally abundant mosquito species could serve as a vector in this area of endemic RVFV circulation. Phylogenetic analysis indicated the identified virus is closely related to two isolates from the earliest outbreaks, which occurred in central South Africa more than 60 years ago, indicating long-term endemicity in the region. Further research is required to understand the eco-epidemiology of RVFV and the vectors responsible for its circulation in the eastern tropical coastal region of southern Africa.

Four-day school week schedules are being adopted with increasing frequency, particularly in rural areas. In this paper we consider the academic implications of students in Oregon attending a four-day school week for the first time when they enter high school. We find 11 th grade math achievement in 0.09 standard deviations lower among four-day school week students, with significant impacts driven by four-day school week students in non-rural settings. We also find a greater number of four-day school week students being classified as chronically absent. Finally, we find reductions in on-time graduation among four-day school week students compared to five-day students.

Bovine brucellosis is a zoonotic disease of global public health and economic importance. South Africa has had a national bovine brucellosis eradication scheme since 1979; however, no published report on elimination progress from any province exists. We analysed laboratory test results of all cattle herds participating in the Gauteng Provincial Veterinary Services' eradication scheme between 2013 and 2018. Herd reactor status and within-herd seroprevalence, modelled using mixed-effects logistic and negative binomial regression models, respectively, showed no significant change over the period. However, provincial State Vet Areas, Randfontein (OR = 1.6; 95% CI: 1.2-2.1; p < 0.001) and Germiston (OR = 1.9; 95% CI: 1.5-2.5, p = 0.008) had higher odds of reactor herds than the Pretoria Area and within-herd prevalence count ratios for these areas were 1.5-fold greater than the Pretoria State Vet Area ( p < 0.001). Reactor herds were associated with increased herd size ( p < 0.001) and larger herd sizes were associated with lower within-herd prevalence ( p < 0.001). Despite no evidence of significant progress toward bovine brucellosis elimination in Gauteng province, variability in bovine brucellosis prevalence between State Vet Areas exists. A public health and farmer-supported strategy of ongoing district-based surveillance and cattle vaccination targeting small- to medium-sized herds combined with compulsory test and slaughter of reactors in larger herds is recommended for the province.

Background: Four-day school week (FDSW) use has increased substantially among US districts in recent years, but limited data exist on health impacts of this school schedule. This study examined associations of reduced school exposure via FDSWs with adolescent health and risk behaviors, obesity, and food security., Methods: Self-report data from 8th- and 11th-grade students from the Oregon Healthy Teens survey across 5 survey years (odd years 2007-2015, total N = 91,860-104,108 respondents depending on the survey question) were linked to a FDSW indicator. Regression analyses controlling for student and school characteristics compared outcomes between students in 4- and 5-day schools overall (without school fixed effects) and outcomes associated with switching to a FDSW (with school fixed effects)., Results: When controlling for multiple student- and school-level factors, we observed adolescents in FDSW schools report they consume sugar sweetened beverages more frequently and water less frequently, have access to fewer days of physical education, are more likely to be food insecure, and are more likely to report the use of any drugs and specifically marijuana than 5-day school week students., Conclusions: Limiting exposure to the school environment via FDSWs may impact adolescent health behaviors, including diet, physical activity, and drug use., (© 2021 The Authors. Journal of School Health published by Wiley Periodicals LLC on behalf of American School Health Association.)

In South Africa, the prevalence of cattle handler exposure to Brucella on cattle farms is unknown and risk factors and cattle symptoms associated with infected cattle herds are unavailable. To address this gap, a case-control study of cattle herds was conducted in Gauteng province and farm workers and veterinary officials were tested for exposure to Brucella . Seroprevalence amongst farm workers exposed to case herds ranged from 4.0% (BrucellaCapt ® ) to 16.7% (IgG ELISA ® ), compared to those exposed to control herds, where seroprevalence ranged from 1.9% (BrucellaCapt ® ) to 5.7% (IgG ELISA ® ). Seroprevalence amongst veterinary officials was significantly greater compared to farm workers exposed to case herds for the outcome RBT+ IgM- IgG+ (OR = 11.1, 95% CI: 2.5-49.9, p = 0.002) and RBT- IgM- IgG+ (OR = 6.3, 95% CI: 2.3-17.3, p < 0.001). Risk factors associated with being an infected herd were: being a government-sponsored farm vs. private farm (OR 4.0; 95% CI: 1.4-11.3; p = 0.009), beef vs. dairy herd (OR 7.9; 95% CI: 1.4-44.9; p = 0.020), open vs. closed herd (OR 3.3; 95% CI: 1.1-10.4; p = 0.038) and the presence of antelope on the farm (OR 29.4; 95% CI: 4.0-218.2; p = 0.001). Abortions (OR = 5.1; 95% CI: 2.0-13.3; p < 0.001), weak calves in the herd (OR = 8.0; 95% CI: 2.6-24.4; p < 0.001), reduction in number of calves born (OR = 9.0; 95% CI: 2.1-43.6; p < 0.001), reduction in conception rate (OR = 3.9; 95% CI: 0.8-18.3; p = 0.046), hygromas in cattle ( p = 0.011) and farmers reporting brucellosis-like symptoms in their farm workers or in him/herself (OR = 3.4; 95% CI: 1.3-8.7; p = 0.006) were more likely to be associated with Brucella infected herds than control herds. This evidence can be used in strategic planning to protect both human and herd health.

Brucellosis in humans is under-detected and underreported in sub-Saharan Africa. Risk factors associated with Brucella infection and health seeking behaviour in response to brucellosis-like symptoms, amongst cattle farm workers and veterinary officials in South Africa, are unknown. Farm workers and veterinary officials ( N = 230) were screened for brucellosis using commercial Rose Bengal Test (RBT ® ), IgM Enzyme-linked Immunoassay (ELISA) ® , IgG ELISA ® and the BrucellaCapt ® test. Knowledge of brucellosis and risk factors for exposure to Brucella were also investigated. Seroprevalence varied according to test used: 10.1% (RBT ® ), 20.9% (IgG ELISA ® ) and 6.5% (BrucellaCapt ® ). Only 22.2% (6/27) of veterinary officials opt to visit a clinic, doctor, or hospital in response to self-experienced brucellosis-like symptoms, compared to 74.9% (152/203) of farm workers ( p < 0.001). Of the BrucellaCapt ® seropositive participants, 53% (7/15) did not visit a clinic in response to brucellosis-like symptoms. Weak evidence of an association between the handling of afterbirth or placenta and infection of a short evolution (RBT ® , IgM ELISA ® and IgG ELISA ® seropositive) was found (OR = 8.9, 95% CI: 1.0-81.1, p = 0.052), and strong evidence of an association between this outcome and the slaughter of cattle (OR = 5.3, 95% CI: 1.4-19.6, p = 0.013). There was strong evidence of a positive association between inactive/resolved infection and veterinary officials vs. farm workers exposed to seropositive herds (OR = 7.0, 95% CI: 2.4-20.2, p < 0.001), with a simultaneous negative association with the handling of afterbirth or placenta (OR = 3.9, 95% CI: 1.3-11.3, p = 0.012). Findings suggest a proportion of undetected clinical cases of brucellosis amongst workers on cattle farms in Gauteng.

Crimean-Congo haemorrhagic fever (CCHF) is a severe tick-borne viral zoonosis endemic to parts of Africa, Europe, the Middle East and Central Asia. Human cases are reported annually in South Africa, with a 25% case fatality rate since the first case was recognized in 1981. We investigated CCHF virus (CCHFV) seroprevalence and risk factors associated with infection in cattle and humans, and the presence of CCHFV in Hyalomma spp. ticks in central South Africa in 2017-18. CCHFV IgG seroprevalence was 74.2% (95%CI: 64.2-82.1%) in 700 cattle and 3.9% (95%CI: 2.6-5.8%) in 541 farm and wildlife workers. No veterinary personnel (117) or abattoir workers (382) were seropositive. The prevalence of CCHFV RNA was significantly higher in Hyalomma truncatum (1.6%) than in H. rufipes (0.2%) (P = 0.002). Seroprevalence in cattle increased with age and was greater in animals on which ticks were found. Seroprevalence in cattle also showed significant geographic variation. Seroprevalence in humans increased with age and was greater in workers who handled livestock for injection and collection of samples. Our findings support previous evidence of widespread high CCHFV seroprevalence in cattle and show significant occupational exposure amongst farm and wildlife workers. Our seroprevalence estimate suggests that CCHFV infections are five times more frequent than the 215 confirmed CCHF cases diagnosed in South Africa in the last four decades (1981-2019). With many cases undiagnosed, the potential seriousness of CCHF in people, and the lack of an effective vaccine or treatment, there is a need to improve public health awareness, prevention and disease control., Competing Interests: The authors have declared that no competing interests exist.

This study aimed to consolidate current knowledge of wildlife brucellosis in Africa and to analyse available predictors of infection. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were followed. Information on species, test used, test results, area, rainfall, livestock and wildlife contact and year of study were extracted. This systematic review revealed 42 prevalence studies, nine disease control articles and six articles on epidemiology. Brucella abortus, Brucella melitensis, Brucella inopinata and Brucella suis were reported in wildlife. The prevalence studies revealed serological evidence of brucellosis in buffalo, antelope (positive in 14/28 species), carnivores (4/12) and other species (7/20) over the last five decades. Buffalo populations were more likely to be infected and had a higher seroprevalence than other species; the pooled seroprevalence was 13.7% (95% CI 10.3-17.3%) in buffalo, 7.1% (95% CI 1.1-15.5%) in carnivores and 2.1% (95% CI 0.1-4.9%) in antelope. Wildlife in high rainfall areas (≥ 800 mm) were more likely to be infected, and infected populations showed higher seroprevalence in high rainfall areas and in studies published after 2000. Domestic animal contact was associated with increased seroprevalence in antelope and carnivore species, but not in buffalo, supporting the hypothesis that buffalo may be a reservoir species.

Q fever is a neglected zoonosis in South Africa, causing significant losses in livestock and game animals through reproductive disorders. However, there are limited studies on the extent of Coxiella burnetii infections in livestock in South Africa. Further, there is also lack of knowledge about the types of C. burnetii strains that are currently circulating in the country. Therefore, a cross-sectional, abattoir-based study was conducted to determine the seroprevalence of C. burnetii and associated risk factors, and to characterize C. burnetii strains from slaughter livestock at red meat abattoirs in Gauteng, South Africa. Of the 507 animals tested, 6.9% (95% CI: 4.9-9.5%) were positive for antibodies against C. burnetii . The seroprevalence was 9.4% (31/331) in cattle, 4.3% (3/69) in sheep, and 0.9% (1/107) in pigs. Out of the 63 tissue samples from 35 seropositive animals including material from two sheep aborted fetuses from Mangaung district (Free State province), 12.7% (8/63) tested positive by IS 1111 PCR. Genotyping of the eight PCR-positive tissues from eight animals by MLVA revealed two novel genotypes, not available in Coxiella MLVA databases. It is concluded that slaughter animals pose a risk of exposing abattoir and farm workers to C. burnetii in South Africa.

Although West Nile virus (WNV) is endemic to South Africa (RSA), it has only become recognized as a significant cause of neurological disease in humans and horses locally in the past 2 decades, as it emerged globally. This article describes the epidemiological and clinical presentation of WNV in horses across RSA during 2016-2017. In total, 54 WNV-positive cases were identified by passive surveillance in horses with febrile and/or neurological signs at the Centre for Viral Zoonoses, University of Pretoria. They were followed up and compared to 120 randomly selected WNV-negative controls with the same case definition and during the same time period. Of the WNV-positive cases, 52% had fever, 92% displayed neurological signs, and 39% experienced mortality. Cases occurred mostly in WNV-unvaccinated horses <5 years old, during late summer and autumn after heavy rain, in the temperate to warm eastern parts of RSA. WNV-positive cases that had only neurological signs without fever were more likely to die. In the multivariable analysis, the odds of WNV infection were associated with season (late summer), higher altitude, more highly purebred animals, younger age, and failure to vaccinate against WNV. Vaccination is currently the most effective prophylactic measure to reduce WNV morbidity and mortality in horses.

The southern ground-hornbill ( Bucorvus leadbeateri ; hereafter SGH) is endangered in South Africa, Namibia, and Swaziland. Through a conservation program established in South Africa by the Mabula Ground Hornbill Project, wild populations are being re-established by the reintroduction of captive-reared birds. The SGH is susceptible to infection with avian avulavirus 1, which causes Newcastle disease (ND). Four different vaccines to protect against ND were administered through various vaccination schedules and evaluated by serologic monitoring to assess the efficiency and safety of various combinations of vaccines (live versus inactivated/killed), vaccine strains (Ulster strain, live; Avivac Cellimune, live; VG/GA strain, live; and Avivac Struvac, killed), and administration routes (intraocular versus subcutaneous versus intramuscular injection versus oral). We vaccinated 75 individuals and evaluated antibody titers in 53 individuals (24 juveniles, 13 subadults, and 16 adult SGH; 26 males and 27 females) over a period of 9 years. Antibody titers to avian avulavirus 1 in sera were monitored by a hemagglutination inhibition test. Protective titers were generated with 3/6 vaccine regimes tested in the SGH. The highest vaccine titers were established in birds vaccinated with the Ulster strain in the conjunctiva and followed with an intramuscular Struvac injection (mean log 2 titer 8.6 ± 2.6) booster. Our aim was 1) to assess whether optimal vaccination protocols could be developed and 2) to then be able, by oral administration, to remove the need to recapture free-roaming, reintroduced birds to administer the initial vaccine or booster, thus remove the threat or mortality associated with ND to this endangered avian species in both captive birds and birds released back into the wild.

The histological effect on the felid uterus of sterilization, via ovariectomy or salpingectomy, is currently unknown. To investigate the association of ovariectomy or salpingectomy with uterine health, it is first necessary to establish if changes are distributed evenly throughout the uterus. Both laparoscopic ovariectomy and salpingectomy with concurrent sampling of the tip of the uterine horn are possible in the cheetah. Currently accepted practice for histopathological screening of the uterus utilizes four biopsy samples. It is not known whether this method accurately reflects the status of the entire uterus. In this study we histologically examined the uteri of six older cheetahs (one 7-year-old and five 10-10.5-year-old animals) via 21 tissue samples (three samples from seven different anatomical regions) per cheetah to determine overall uterine health. Although no defined lesions were detected, mild endometrial gland dilation, assumed to be of no functional consequence, was observed in multiple samples. The odds of observing this dilation was lowest in the uterine body and progressively increased in a cranial direction, being significantly higher at the tip of the uterine horns (OR = 11.5; 95% CI, 2.0-65.1; p = 0.006). This supported the reliability of sampling the tip of the uterine horn to screen for endometrial gland dilation., (© 2020 Wiley Periodicals LLC.)

An African horse sickness (AHS) outbreak occurred in March and April 2016 in the controlled area of South Africa. This extended an existing trade suspension of live equids from South Africa to the European Union. In the post-outbreak period ongoing passive and active surveillance, the latter in the form of monthly sentinel surveillance and a stand-alone freedom from disease survey in March 2017, took place. We describe a stochastic scenario tree analysis of these surveillance components for 24 months, starting July 2016, in three distinct geographic areas of the controlled area. Given that AHS was not detected, the probability of being free from AHS was between 98.3% and 99.8% assuming that, if it were present, it would have a prevalence of at least one infected animal in 1% of herds. This high level of freedom probability had been attained in all three areas within the first 9 months of the 2-year period. The primary driver of surveillance outcomes was the passive surveillance component. Active surveillance components contributed minimally (<0.2%) to the final probability of freedom. Sensitivity analysis showed that the probability of infected horses showing clinical signs was an important parameter influencing the system surveillance sensitivity. The monthly probability of disease introduction needed to be increased to 20% and greater to decrease the overall probability of freedom to below 90%. Current global standards require a 2-year post-incursion period of AHS freedom before re-evaluation of free zone status. Our findings show that the length of this period could be decreased if adequately sensitive surveillance is performed. In order to comply with international standards, active surveillance will remain a component of AHS surveillance in South Africa. Passive surveillance, however, can provide substantial evidence supporting AHS freedom status declarations, and further investment in this surveillance activity would be beneficial., (© 2020 Blackwell Verlag GmbH.)

Background: Use of 4-day school weeks (FDSWs) as a cost-saving strategy has increased substantially as many US school districts face funding declines. However, the impacts of FDSWs on physical activity exposure and related outcomes are unknown. This study examined physical education (PE) exposure and childhood obesity prevalence in 4- versus 5-day Oregon schools; the authors hypothesized lower PE exposure and higher obesity in FDSW schools, given reduced school environment exposure., Methods: The authors utilized existing data from Oregon to compare 4- versus 5-day models: t tests compared mean school-level factors (PE exposure, time in school, enrollment, and demographics) and complex samples weighted t tests compared mean child-level obesity data for a state representative sample of first to third graders (N = 4625)., Results: Enrollment, time in school, and student-teacher ratio were significantly lower in FDSW schools. FDSW schools provided significantly more PE, both in minutes (120 vs 101 min/wk in 4- vs 5-d schools, P < .01) and relative to total time in school (6.9% vs 5.0%, P < .0001). Obesity prevalence did not differ significantly between school models., Conclusion: Greater PE exposure in FDSW schools was observed, and it remains unknown whether differences in PE exposure contributed to obesity prevalence in this sample of students. Efforts to better understand how FDSWs impact physical activity, obesity risk, and related factors are needed.

Rift Valley fever (RVF) is a zoonotic viral disease of domestic ruminants in Africa and the Arabian Peninsula caused by a mosquito-borne Phlebovirus. Outbreaks in livestock and humans occur after heavy rains favour breeding of vectors, and the virus is thought to survive dry seasons in the eggs of floodwater-breeding aedine mosquitoes. We recently found high seroconversion rates to RVF virus (RVFV) in cattle and goats, in the absence of outbreaks, in far northern KwaZulu-Natal (KZN), South Africa. Here, we report the prevalence of, and factors associated with, neutralizing antibodies to RVFV in 326 sera collected opportunistically from nyala (Tragelaphus angasii) and impala (Aepyceros melampus) culled during 2016-2018 in two nature reserves in the same area. The overall seroprevalence of RVFV, determined using the serum neutralization test, was 35.0% (114/326; 95%CI: 29.8%-40.4%) and tended to be higher in Ndumo Game Reserve (11/20; 55.0%; 95%CI: 31.5%-76.9%) than in Tembe Elephant Park (103/306; 33.6%; 95%CI: 28.4%-39.3%) (p = .087). The presence of antibodies in juveniles (6/21; 28.6%; 95%CI: 11.3%-52.2%) and sub-adults (13/65; 20.0%; 95%CI: 11.1%-37.8%) confirmed that infections had occurred at least until 2016, well after the 2008-2011 RVF outbreaks in South Africa. Odds of seropositivity was higher in adults than in sub-adults (OR = 3.98; 95%CI: 1.83-8.67; p = .001), in males than in females (OR = 2.66; 95%CI: 1.51-4.68; p = .001) and in animals collected ≤2 km from a swamp or floodplain compared with those collected further away (OR = 3.30; 95%CI: 1.70-6.38; p < .001). Under similar ecological conditions, domestic and wild ruminants may play a similar role in maintenance of RVFV circulation and either or both may serve as the mammalian host in a vector-host reservoir system. The study confirms the recent circulation of RVFV in the tropical coastal plain of northern KZN, providing the basis for investigation of factors affecting virus circulation and the role of wildlife in RVF epidemiology., (© 2020 Blackwell Verlag GmbH.)

A cross sectional sero-epidemiological study was conducted on cattle in a communal farming area adjacent to Kruger National Park at a wildlife-livestock interface in South Africa. A total of 184 cattle were screened for exposure to 5 abortifacient or zoonotic pathogens, namely Coxiella burnetii , Toxoplasma gondii , Chlamydophila abortus , Neospora caninum , and Rift Valley fever virus (RVFV) using enzyme-linked immunosorbent assays. In addition, the virus neutralization test was used to confirm the presence of antibodies to RVFV. The seroprevalence of C. burnetii , T. gondii , C. abortus , N. caninum , and RVFV antibodies was 38.0%, 32.6%, 20.7%, 1.6%, and 0.5%, respectively, and varied between locations ( p  < 0.001). Seroprevalence of C. burnetii and T. gondii was highly clustered by location (intraclass correlation coefficient [ICC] = 0.57), and that of C. abortus moderately so (ICC = 0.11). Seroprevalence was not associated with sex or age for any pathogen, except for C. abortus , for which seroprevalence was positively associated with age ( p  = 0.01). The predominant mixed infections were C. burnetii and T. gondii (15.2%) and C. burnetii , T. gondii , and C. abortus (13.0%). The serological detection of the five abortifacient pathogens in cattle indicates the potential for economic losses to livestock farmers, health impacts to domestic animals, transmission across the livestock-wildlife interface, and the risk of zoonotic transmission. This is the first documentation of T. gondii infection in cattle in South Africa, while exposure to C. burnetii , C. abortus , and N. caninum infections is being reported for the first time in cattle in a wildlife-livestock interface in the country.

Rift Valley fever (RVF) is a mosquito-borne viral zoonosis showing complex epidemiological patterns that are poorly understood in South Africa. Large outbreaks occur in the central interior at long, irregular intervals, most recently in 2010-2011; however, the level of herd immunity of ruminant livestock, a key determinant of outbreaks, is unknown. During 2015-2016 a cross-sectional study on 234 randomly-selected farms investigated the prevalence, patterns of, and factors associated with, antibodies to RVF virus (RVFV) in livestock in an area heavily affected by that outbreak. A RVFV inhibition ELISA was used to screen 977 cattle, 1,549 sheep and 523 goats and information on potential risk factors was collected using a comprehensive questionnaire. The estimated RVFV seroprevalence, adjusted for survey design, was 42.9% in cattle, 28.0% in sheep and 9.3% in goats, showing a high degree of farm-level clustering. Seroprevalence increased with age and was higher on private vs. communal land, on farms with seasonal pans (temporary, shallow wetlands) and perennial rivers and in recently vaccinated animals. Seropositivity amongst unvaccinated animals born after the last outbreak indicates likely viral circulation during the post-epidemic period. The current level of herd immunity in livestock may be insufficient to prevent another large outbreak, should suitable conditions recur.

Abstract: A cross-sectional study was conducted to determine the prevalence of and factors associated with Shiga toxin-producing Escherichia coli (STEC) in raw beef and ready-to-eat (RTE) beef products sold in 31 retail outlets in Pretoria, South Africa, and nearby areas. A total of 463 beef and RTE samples were screened for four STEC virulence genes (stx1, stx2, eaeA, and hlyA) and seven O-serogroups (O113, O157, O26, O91, O145, O111, and O103) with a multiplex PCR assay. The total aerobic plate count (TAPC) per gram was also determined. A total of 38 STEC isolates were recovered and characterized by conventional PCR assay and serotyping. The overall prevalence of STEC in the beef and RTE samples tested was 16.4% (76 of 463 samples; 95% confidence interval, 13 to 20%). The prevalence of STEC differed significantly by product type (P < 0.0001), with the highest prevalence (35%) detected in boerewors (spicy sausage). The STEC prevalences in minced beef, brisket, RTE cold beef, and biltong were 18, 13, 9, and 5%, respectively. The most frequently detected stx gene was stx2 (13%), and STEC serogroups from recovered isolates were detected at the following prevalences: O2, 15%; O8, 12%; O13, 15%; O20, 8%; O24, 3%; O39, 3%; O41, 8%; O71, 3%; O76, 3%; O150, 12%; and O174, 3%. A high proportion (77%) of the samples had TAPCs that exceeded the South African microbiological standards for meat export (5.0 log CFU/g). The prevalence of O157 STEC (16%) and the diversity of non-O157 STEC serogroups found in five common beef-based products from retail outlets in South Africa suggest exposure of raw beef and beef products to multiple contamination sources during carcass processing and/or cutting and handling at retail outlets. These data provide direct estimates of the potential health risk to consumers from undercooked contaminated products and indicate the need to improve sanitary practices during slaughter and processing of beef and beef-based RTE products. A risk-based surveillance system for STEC may be needed., (Copyright ©, International Association for Food Protection.)

An African horse sickness (AHS) outbreak occurred in South Africa's AHS controlled area in autumn 2016. A freedom from disease survey was performed to establish the likelihood of ongoing circulation of the associated virus during the same period the following year. A single-stage surveillance strategy was employed with a population-level design prevalence of 1% to establish a survey population sensitivity of 95% (probability that one or more positive horses would be detected if AHS was present at a prevalence greater than or equal to the design prevalence). In March 2017, a total of 262 randomly selected horses from 51 herds were sampled from the 2016 outbreak containment zone. Three within-herd and herd-level design prevalence scenarios were used in evaluating the post-survey probability of freedom. Depending on the underlying design prevalence scenarios, effectively ranging between 0.8% and 6.4%, and the use of informed or uninformed priors, the probability of freedom derived from this surveillance ranged between 73.1% and 99.9% (uninformed prior) and between 96.6% and 100% (informed prior). Based on the results, the authors conclude that it is unlikely that the 2016 AHS virus was still circulating in the autumn of 2017 in the 2016 outbreak containment zone. The ability to perform freedom from disease surveys, and also to include risk-based methods, in the AHS controlled area of South Africa is influenced by the changing underlying population at risk and the high level of vaccination coverage in the horse population. Ongoing census post-outbreak must be undertaken to maintain a valid sampling frame for future surveillance activity. The seasonality of AHS, the restricted AHS vaccination period and the inability to easily differentiate infected from vaccinated animals by laboratory testing impact the ability to perform a freedom from disease survey for AHS in the 12 months following an outbreak in the controlled area., (© 2019 Blackwell Verlag GmbH.)

Babesia rossi causes the most severe clinical disease in dogs of all the babesia parasites. We included 320 naturally-infected dogs that presented for care at the Onderstepoort Veterinary Academic Hospital between 2006 and 2016. All dogs had mono-infections confirmed by multiplex PCR. The data allowed more accurate clinical classification of the disease and identified parameters that were associated with disease severity and death. Odds ratios for dying were significant (P < 0.05) for increased band neutrophil count, collapse at presentation; presence of cerebral signs; hypoglycaemia; hyperlactatemia; high urea, high creatinine; hyperbilirubinaemia; hypercortisolaemia; and hypothyroxinaemia. Joint component analysis confirmed that the variables with significant odds ratios grouped together with death. Yet, multivariate logistic regression was unable to identify a group of significant independent predictors of death. Receiver Operator Characteristic curves indicated that low total thyroid hormone, high bilirubin, high serum urea and high cortisol concentrations were the variables with the highest sensitivity and specificity for death. These data provide both the clinician and researcher with a set of easily-measured laboratory and clinical assessments to classify cases into those that are uncomplicated and those that are complicated. The disease is complex and multisystemic and probably involves mechanisms more proximal in the pathogenesis than those that have been evaluated., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Lumpy skin disease (LSD) is an important transboundary animal disease of cattle with significant economic impact because of the implications for international trade in live animals and animal products. LSD is caused by a Capripoxvirus, LSD virus (LSDV), and results in extensive hide and udder damage, fever and pneumonia. LSDV can be shed in semen of infected bulls for prolonged periods and transmitted venereally to cows at high doses. This study examined the effects of LSDV in frozen-thawed semen on in vitro embryo production parameters, including viral status of media and resulting embryos. Bovine oocytes were harvested from abattoir-collected ovaries and split into three experimental groups. After maturation, the oocytes were fertilized in vitro with frozen-thawed semen spiked with a high (HD) or a lower (LD) dose of LSDV, or with LSDV-free semen (control). Following day 7 and day 8 blastocyst evaluation, PCR and virus isolation were performed on all embryonic structures. After completing sufficient replicates to reach 1,000 inseminated oocytes, further in vitro fertilization (IVF) runs were performed to provide material for electron microscopy (EM) and embryo washing procedures. Overall, in vitro embryo yield was significantly reduced by the presence of LSDV in frozen-thawed semen, irrespective of viral dose. When semen with a lower viral dose was used, significantly lower oocyte cleavage rates were observed. LSDV could be detected in fertilization media and all embryo structures, when higher doses of LSDV were present in the frozen-thawed semen used for IVF. Electron microscopy demonstrated LSDV virions inside blastocysts. Following the International Embryo Transfer Society washing procedure resulted in embryos free of viral DNA; however, this may be attributable to a sampling dilution effect and should be interpreted with caution. Further research is required to better quantify the risk of LSDV transmission via assisted reproductive procedures., (© 2019 Blackwell Verlag GmbH.)

High blood urea nitrogen (BUN) concentration decreases fertility in ruminants. Nguni cattle are reported to maintain BUN concentration more efficiently than other beef breeds. Our objectives were to determine if BUN concentration differed between Nguni and Hereford cows exposed to a high protein ration, and if breed or BUN and serum protein concentrations at the time of oocyte pick-up affected oocyte quantity, quality, and viability. Twelve Nguni and 10 Hereford cows were randomized into high or normal BUN-inducing diets in a crossover design. Ultrasound-guided oocyte pick-up was performed twice weekly; oocytes were counted, visually graded and the viable oocytes were pooled by treatment and breed for in vitro maturation, fertilization, and culture. Nguni cows on the highest protein ration achieved lower mean BUN concentration than Herefords (P < 0.05), and Nguni cows reached BUN concentrations above 20 mg/dL less frequently than Herefords (P = 0.03). Donor BUN concentration above 20 mg/dL at the time of oocyte pick-up, but not breed, independently decreased the number of good quality oocytes harvested. Increasing weighted mean serum albumin of donor cows was independently associated with the number of oocytes that cleaved by day 2 and that reached morula stage by day 7 (P = 0.01). In conclusion, Nguni cows reached the critical threshold of 20 mg/dL BUN less frequently than Herefords; BUN of donor cows above 20 mg/dL negatively affected visual oocyte quality independent of breed, and increasing serum albumin of donor cows improved viability of bovine oocytes.

Osteoarthritis is a common cause of lameness in horses, resulting in poor performance. Intra-articular platelet-rich plasma can deliver a collection of bioactive molecules, such as autologous growth factors and proteins involved in the quality of tissue repair. Horses (n=5) with osteoarthritis affecting antebrachiocarpal, middle carpal or metacarpophalangeal joints, and horses (n=5) without osteoarthritis of the corresponding joints (radiographically free of osteoarthritis), were used for the production of platelet-rich plasma which was subsequently injected into selected joints. Clinical and synovial fluid changes after intra-articular injection of platelet-rich plasma as well as synovial platelet-derived growth factor-BB and transforming growth factor-beta 1 concentration changes were evaluated in these joints and compared between normal joints and joints with osteoarthritis. A gravity filtration system produced a moderately concentrated platelet-rich plasma, representing a 4.7-fold increase in baseline platelet concentration. The synovial effusion score was significantly different between the control joints and joints with osteoarthritis on Day 0 with a higher score in the group with osteoarthritis. Within the control group, the synovial effusion score was significantly higher on Days 1 and 2 compared to Day 0. For both groups, the synovial fluid nucleated cell count, predominantly intact neutrophils, was significantly increased on Days 1 and 2, with no significant difference between groups. The mean synovial platelet-derived growth factor-BB and transforming growth factor-beta 1 concentrations were increased for both groups but significantly lowered in the group with osteoarthritis on Day 1 compared to normal joints. Concentrations for platelet-derived growth factor-BB remained unchanged on Day 5, compared to Day 1, with no significant difference between groups. In conclusion, intra-articular treatment with platelet-rich plasma resulted in increased synovial growth factor concentrations in joints but with lower concentrations in joints with osteoarthritis. A transient inflammatory reaction was seen both clinically as an increase in synovial effusion and cytologically in both normal joints and joints with osteoarthritis.

Background: Rift Valley fever (RVF) is a mosquito-borne zoonotic disease characterized in South Africa by large epidemics amongst ruminant livestock at very long, irregular intervals, mainly in the central interior. However, the presence and patterns of occurrence of the virus in the eastern parts of the country are poorly known. This study aimed to detect the presence of RVF virus (RVFV) in cattle and goats in far northern KwaZulu-Natal province and to estimate the prevalence of antibodies to the virus and the incidence rate of seroconversion., Methodology: Cross-sectional studies were performed in communally farmed cattle (n = 423) and goats (n = 104), followed by longitudinal follow-up of seronegative livestock (n = 253) 14 times over 24 months, representing 160.3 animal-years at risk. Exposure to RVFV was assessed using an IgG sandwich ELISA and a serum neutralization test (SNT) and seroconversion was assessed using SNT. Incidence density was estimated and compared using multivariable Poisson models and hazard of seroconversion was estimated over time., Principal Findings: Initial overall seroprevalence was 34.0% (95%CI: 29.5-38.8%) in cattle and 31.7% (95%CI: 22.9-41.6%) in goats, varying by locality from 18-54%. Seroconversions to RVFV based on SNT were detected throughout the year, with the incidence rate peaking during the high rainfall months of January to March, and differed considerably between years. Overall seroconversion rate in cattle was 0.59 per animal-year (95% CI: 0.46-0.75) and in goats it was 0.41 per animal-year (95% CI: 0.25-0.64), varying significantly over short distances., Conclusions/significance: The high seroprevalence in all age groups and evidence of year-round viral circulation provide evidence for a hyperendemic situation in the study area. This is the first study to directly estimate infection rate of RVFV in livestock in an endemic area in the absence of reported outbreaks and provides the basis for further investigation of factors affecting viral circulation and mechanisms for virus survival during interepidemic periods., Competing Interests: The authors have declared that no competing interests exist.

An outbreak of African horse sickness (AHS) caused by AHS virus type 1 occurred within the South African AHS surveillance zone during April and May 2016. The index case was detected by a private veterinarian through passive surveillance. There were 21 cases in total, which is relatively low compared to case totals during prior AHS outbreaks in the same region (and of the same AHS virus type) in 2004, 2011 and 2014. The affected proportion of horses on affected properties was 0.07 (95% CI 0.04, 0.11). Weather conditions were conducive to high midge activity immediately prior to the outbreak but midge numbers decreased rapidly with the advent of winter. The outbreak was localized, with 18 of the 21 cases occurring within 8 km of the index property and the three remaining cases on two properties within 21 km of the index property, with direction of spread consistent with wind-borne dispersion of infected midges. Control measures included implementation of a containment zone with movement restrictions on equids. The outbreak was attributed to a reversion to virulence of a live attenuated vaccine used extensively in South Africa. Outbreaks in the AHS control zones have a major detrimental impact on the direct export of horses from South Africa, notably to the European Union., (© 2018 Blackwell Verlag GmbH.)

Rift Valley fever (RVF) is a re-emerging arboviral disease of public health and veterinary importance in Africa and the Arabian Peninsula. Major RVF epidemics were documented in South Africa in 1950⁻1951, 1974⁻1975, and 2010⁻2011. The number of individuals infected during these outbreaks has, however, not been accurately estimated. A total of 823 people in close occupational contact with livestock were interviewed and sampled over a six-month period in 2015⁻2016 within a 40,000 km² study area encompassing parts of the Free State and Northern Cape provinces that were affected during the 2010⁻2011 outbreak. Seroprevalence of RVF virus (RVFV) was 9.1% (95% Confidence Interval (CI95%): 7.2⁻11.5%) in people working or residing on livestock or game farms and 8.0% in veterinary professionals. The highest seroprevalence (SP = 15.4%; CI95%: 11.4⁻20.3%) was detected in older age groups (≥40 years old) that had experienced more than one known large epidemic compared to the younger participants (SP = 4.3%; CI95%: 2.6⁻7.3%). The highest seroprevalence was in addition found in people who injected animals, collected blood samples (Odds ratio (OR) = 2.3; CI95%: 1.0⁻5.3), slaughtered animals (OR = 3.9; CI95%: 1.2⁻12.9) and consumed meat from an animal found dead (OR = 3.1; CI95%: 1.5⁻6.6), or worked on farms with dams for water storage (OR = 2.7; CI95%: 1.0⁻6.9). We estimated the number of historical RVFV infections of farm staff in the study area to be most likely 3849 and 95% credible interval between 2635 and 5374 based on seroprevalence of 9.1% and national census data. We conclude that human RVF cases were highly underdiagnosed and heterogeneously distributed. Improving precautions during injection, sample collection, slaughtering, and meat processing for consumption, and using personal protective equipment during outbreaks, could lower the risk of RVFV infection., Competing Interests: The authors declare no conflict of interest.

Adequate nutrition is required for maintenance of normal reproduction in cattle. Albumin, the best marker and fundamental part of nutrition, most abundant plasma protein and major component of fetal bovine serum, is the best predictor of malnourishment in South African cattle. The aim of this study was to determine if serum albumin concentrations of donor cows predict the developmental competence of oocytes, and if additional protein supplementation of the in vitro culture media improves embryo outcomes in oocytes from cows with inadequate serum albumin concentrations. Oocytes (n = 1024) were recovered from donors with inadequate (≤35.9 g/L), or adequate serum albumin concentrations (≥36.0 g/L). Four hundred and sixty oocytes originated from cows with inadequate serum albumin and 564 from cows with adequate serum albumin. Oocytes of these cohorts were randomly allocated to a control and supplemented fetal bovine serum in vitro embryo culture protocol. Multiple linear, logistic and Poisson regression analyses were performed to estimate the effects of different covariates on linear, binary and count data respectively. Mixed effects Poisson regression was performed for the number of oocytes that developed into blastocysts by the seventh day of culture. Adequate serum albumin concentration of donor cows independently resulted in 46% increased blastocyst formation in the control protocol (P = 0.02). Although fetal bovine serum supplementation of the culture protocol did not affect blastocyst formation in oocytes originating from cows with inadequate serum albumin, it independently reduced blastocyst formation by 30% in oocytes originating from cows with adequate serum albumin (P = 0.02). Other independent predictors of blastocyst outcome included higher serum urea nitrogen, lower beta (β)-hydroxybutyric acid concentrations and lower fat classification of donor cows. It is concluded that adequate serum albumin of donor cows is a significant predictor of developmental competence of oocytes, and that in vitro supplementation of fetal bovine serum does not improve developmental competence of oocytes and can lead to negative blastocyst outcomes. Further research is required to determine optimal protein supplementation for oocytes originating from inadequately nourished cows., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The prevalence of nasal carrier status of methicillin-resistant Staphylococcus aureus (MRSA) in pigs has been described elsewhere, but is unknown in South Africa. To address concerns that exist regarding the zoonotic risk that carriers pose to workers, the herd-level prevalence of MRSA was determined among 25 large (> 500 sows) commercial pig herds in South Africa, representing 45% of the large commercial herds in the country. From each herd, the nasal contents of 18 finisher pigs were sampled at the abattoir, pooled into three and selectively cultured to determine the presence of MRSA. A herd was classified as MRSA-positive if one or more of the three pooled samples cultured positive. Three of the 25 herds tested positive for MRSA, equating to a 12% herd prevalence (95% CI: 7% - 23%) among South African commercial piggeries. The prevalence of nasal MRSA carriers among large commercial pig herds in South Africa was low compared to what has been reported elsewhere and suggests a relatively low zoonotic MRSA risk to workers in South African commercial piggeries and abattoirs.

Naltrexone is used to antagonise etorphine immobilisation, but a safe and effective dose for this purpose has not been objectively determined. Eight domestic goats were immobilised with etorphine (0.07 mg/kg) eight times at ≥13 day intervals. Naltrexone at doses of 0.5, 1, 2, 5, 10, 20 and 40 mg/mg etorphine were administered intravenously 17 minutes after etorphine injection. Effectiveness of antagonism was recorded based on recovery and renarcotisation scores and clinical observations. All doses produced rapid recovery to the point of standing (median 59 seconds, range 33-157 seconds), with no significant differences in recovery times (P=0.44). The lower naltrexone doses resulted in renarcotisation in some goats: 4/8 in the 10-mg dose trial, 7/8 in the 5-mg dose trial, and 8/8 in the 2-mg, 1-mg and 0.5-mg dose trials. Lower doses resulted in more severe signs of renarcotisation. Complications of renarcotisation included increased body temperature; this occurred just before signs of renarcotisation and was greater in animals with high renarcotisation scores (P<0.01). The lowest, safest effective naltrexone dose that we used to antagonise etorphine immobilisation was 20 mg/mg etorphine, which produced rapid recovery to standing with no renarcotisation., Competing Interests: Competing interests: None declared., (© British Veterinary Association (unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.)

To investigate cases of acute oxalate nephrosis without evidence of ethylene glycol exposure, archived data and tissues from cheetahs ( Acinonyx jubatus) from North America ( n = 297), southern Africa ( n = 257), and France ( n = 40) were evaluated. Renal and gastrointestinal tract lesions were characterized in a subset of animals with ( n = 100) and without ( n = 165) oxalate crystals at death. Crystals were confirmed as calcium oxalate by Raman spectroscopy in 45 of 47 cheetahs tested. Crystals were present in cheetahs from 3.7 months to 15.9 years old. Cheetahs younger than 1.5 years were less likely to have oxalates than older cheetahs ( P = .034), but young cheetahs with oxalates had more oxalate crystals than older cheetahs ( P < .001). Cheetahs with oxalate crystals were more likely to have renal amyloidosis, interstitial nephritis, or colitis and less likely to have glomerular loop thickening or gastritis than those without oxalates. Crystal number was positively associated with renal tubular necrosis ( P ≤ .001), regeneration ( P = .015), and casts ( P ≤ .001) but inversely associated with glomerulosclerosis, renal amyloidosis, and interstitial nephritis. Crystal number was unrelated to the presence or absence of colitis and was lower in southern African than American and European animals ( P = .01). This study found no evidence that coexisting chronic renal disease (amyloidosis, interstitial nephritis, or glomerulosclerosis), veno-occlusive disease, gastritis, or enterocolitis contributed significantly to oxalate nephrosis. Oxalate-related renal disease should be considered as a potential cause of acute renal failure, especially in young captive cheetahs. The role of location, diet, stress, and genetic predisposition in the pathogenesis of oxalate nephrosis in cheetahs warrants further study.