Environmental factors are common forces driving infectious disease dynamics. We compared interannual and seasonal patterns of anthrax infections in two multihost systems in southern Africa: Etosha National Park, Namibia, and Kruger National Park, South Africa. Using several decades of mortality data from each system, we assessed possible transmission mechanisms behind anthrax dynamics, examining (1) within‐ and between‐species temporal case correlations and (2) associations between anthrax mortalities and environmental factors, specifically rainfall and the Normalized Difference Vegetation Index (NDVI), with empirical dynamic modeling. Anthrax cases in Kruger had wide interannual variation in case numbers, and large outbreaks seemed to follow a roughly decadal cycle. In contrast, outbreaks in Etosha were smaller in magnitude and occurred annually. In Etosha, the host species commonly affected remained consistent over several decades, although plains zebra (Equus quagga) became relatively more dominant. In Kruger, turnover of the main host species occurred after the 1990s, where the previously dominant host species, greater kudu (Tragelaphus strepsiceros), was replaced by impala (Aepyceros melampus). In both parks, anthrax infections showed two seasonal peaks, with each species having only one peak in a year. Zebra, springbok (Antidorcas marsupialis), wildebeest (Connochaetes taurinus), and impala cases peaked in wet seasons, while elephant (Loxodonta africana), kudu, and buffalo (Syncerus caffer) cases peaked in dry seasons. For common host species shared between the two parks, anthrax mortalities peaked in the same season in both systems. Among host species with cases peaking in the same season, anthrax mortalities were mostly synchronized, which implies similar transmission mechanisms or shared sources of exposure. Between seasons, outbreaks in one species may contribute to more cases in another species in the following season. Higher vegetation greenness was associated with more zebra and springbok anthrax mortalities in Etosha but fewer elephant cases in Kruger. These results suggest that host behavioral responses to changing environmental conditions may affect anthrax transmission risk, with differences in transmission mechanisms leading to multihost biseasonal outbreaks. This study reveals the dynamics and potential environmental drivers of anthrax in two savanna systems, providing a better understanding of factors driving biseasonal dynamics and outbreak variation among locations.

Disease monitoring in free-ranging wildlife is a challenge and often relies on passive surveillance. Alternatively, proactive surveillance that relies on the detection of specific antibodies could give more reliable and timely insight into disease presence and prevalence in a population, especially if the evidence of disease occurs below detection thresholds for passive surveillance. Primary binding assays, like the indirect ELISA for antibody detection in wildlife, are hampered by a lack of species-specific conjugates. In this study, we developed anti-kudu (Tragelaphus strepsiceros) and anti-impala (Aepyceros melampus) immunoglobulin-specific conjugates in chickens and compared them to the binding of commercially available protein-G and protein-AG conjugates, using an ELISA-based avidity index. The conjugates were evaluated for cross-reaction with sera from other wild herbivores to assess future use in ELISAs. The developed conjugates had a high avidity of >70% against kudu and impala sera. The commercial conjugates (protein-G and protein-AG) had significantly low relative avidity (50% with the impala and kudu conjugates and <40% with the commercial conjugates. These results demonstrate that species-specific conjugates are important tools for the development and validation of immunoassays in wildlife and for the surveillance of zoonotic agents along the livestock-wildlife-human interface.

The diagnosis of anthrax, a zoonotic disease caused by Bacillus anthracis can be complicated by detection of closely related species. Conventional diagnosis of anthrax involves microscopy, culture identification of bacterial colonies and molecular detection. Genetic markers used are often virulence gene targets such as B. anthracis protective antigen (pagA, also called BAPA, occurring on plasmid pXO1), lethal factor (lef, on pXO1), capsule-encoding capB/C (located on pXO2) as well as chromosomal Ba-1. Combinations of genetic markers using real-time/quantitative polymerase chain reaction (qPCR) are used to confirm B. anthracis from culture but can also be used directly on diagnostic samples to avoid propagation and its associated biorisks and for faster identification. We investigated how the presence of closely related species could complicate anthrax diagnoses with and without culture to standardise the use of genetic markers using qPCR for accurate anthrax diagnosis. Using blood smears from 2012–2020 from wildlife mortalities (n = 1708) in Kruger National Park in South Africa where anthrax is endemic, we contrasted anthrax diagnostic results based on qPCR, microscopy, and culture. From smears, 113/1708 grew bacteria in culture, from which 506 isolates were obtained. Of these isolates, only 24.7% (125 isolates) were positive for B. anthracis based on genetic markers or microscopy. However, among these, merely 4/125 (3.2%) were confirmed B. anthracis isolates (based on morphology, microscopy, and sensitivity testing to penicillin and gamma-phage) from the blood smear, likely due to poor survival of spores on stored smears. This study identified B. cereus sensu lato, which included B. cereus and B. anthracis, Peribacillus spp., and Priestia spp. clusters using gyrB gene in selected bacterial isolates positive for pagA region using BAPA probe. Using qPCR on blood smears, 52.1% (890 samples) tested positive for B. anthracis based on one or a combination of genetic markers which included the 25 positive controls. Notably, the standard lef primer set displayed the lowest specificity and accuracy. The Ba-1+BAPA+lef combination showed 100% specificity, sensitivity, and accuracy. Various marker combinations, such as Ba-1+capB, BAPA+capB, Ba-1+BAPA+capB+lef, and BAPA+lef+capB, all demonstrated 100.0% specificity and 98.7% accuracy, while maintaining a sensitivity of 96.6%. Using Ba-1+BAPA+lef+capB, as well as Ba-1+BAPA+lef with molecular diagnosis accurately detects B. anthracis in the absence of bacterial culture. Systematically combining microscopy and molecular markers holds promise for notably reducing false positives. This significantly enhances the detection and surveillance of diseases like anthrax in southern Africa and beyond and reduces the need for propagation of the bacteria in culture.

Background: Although the rate of emerging infectious diseases that originate in wildlife has been increasing globally in recent decades, there is currently a lack of epidemiological data from wild animals.Methodology: We used serology to determine prior exposure to foot‐and‐mouth disease virus (FMDV), Brucella spp., and Coxiella burnetii and used genetic testing to detect blood‐borne parasitic infections in the genera Ehrlichia, Anaplasma, Theileria, and Babesia from wildlife in two national parks, Kruger National Park (KNP), South Africa, and Etosha National Park (ENP), Namibia. Serum and whole blood samples were obtained from free‐roaming plains zebra (Equus quagga), greater kudu (Tragelaphus strepsiceros), impala (Aepyceros melampus), and blue wildebeest (Connochaetes taurinus). Risk factors (host species, sex, and sampling park) for infection with each pathogen were assessed, as well as the prevalence and distribution of co‐occurring infections.Results: In KNP 13/29 (45%; confidence interval [CI]: 26%–64%) kudus tested positive for FMD, but none of these reacted to SAT serotypes. For brucellosis, seropositive results were obtained for 3/29 (10%; CI: 2%–27%) kudu samples. Antibodies against C. burnetii were detected in 6/29 (21%; CI: 8%–40%) kudus, 14/21 (67%; CI: 43%–85%) impalas, and 18/39 (46%; CI: 30%–63%) zebras. A total of 28/28 kudus tested positive for Theileria spp. (100%; CI: 88%–100%) and 27/28 for Anaplasma/Ehrlichia spp. (96%; CI: 82%–100%), whereas 12/19 impalas (63%) and 2/39 zebra (5%) tested positive for Anaplasma centrale. In ENP, only 1/29 (3%; CI: 0%–18%) wildebeest samples tested positive for FMD. None of the samples tested positive for brucellosis, while C. burnetii antibodies were detected in 26/30 wildebeests (87%; CI: 69%–96%), 16/40 kudus (40%; CI: 25%–57%), and 26/26 plains zebras (100%; CI: 87%–100%). A total of 60% Anaplasma/Ehrlichia spp. and 35% Theileria/Babesia spp. in kudu and 37% wildebeest tested positive to Theileria sp. (sable), 30% to Babesia occultans, and 3%–7% to Anaplasma spp. The seroprevalence of Q fever was significantly higher in ENP, while Brucella spp., Anaplasma, Ehrlichia, Theileria, and Babesia species were significantly higher in KNP. Significant coinfections were also identified.Conclusion: This work provided baseline serological and molecular data on 40+ pathogens in four wildlife species from two national parks in southern Africa.

Fasciolosis causes significant economic losses in commercial cattle herds in South Africa, but its prevalence is unknown in most communal areas. A cross-sectional study was conducted with the aim of determining the occurrence of bovine fasciolosis using three different diagnostic methods in Moretele Local Municipality in Bojanala District, North West Province. Faecal samples were collected from 277 cattle of different breeds, ages, sex and faecal condition scores and examined using the sedimentation technique, quantitative real-time polymerase chain reaction (qPCR) and faecal antigen enzyme-linked immunosorbent assay (coproELISA). All samples were negative for bovine fasciolosis using coproELISA. A total of 73 (26.4%) samples were positive using the qPCR, while 36 were positive using the sedimentation technique, with low faecal egg counts (1 to 20 eggs per gram). The qPCR detected the highest positivity (26.4%, 95% CI 21.3, 32.0) followed by the sedimentation test (13.0%; 95% CI 9.3, 17.5). Location, breed, sex, age and faecal consistency score were not associated with positive qPCR results (p > 0.05). There was also no significant agreement (kappa = −0.011, p = 0.843) between qPCR and the sedimentation technique for the detection of Fasciola spp. The qPCR appeared to be the most sensitive method for detection of Fasciola spp. Further studies are required on the characterisation of Fasciola spp. in communal cattle in South Africa.