Epidemiology of Anaplasma species amongst cattle in Africa from 1970 to 2022: A systematic review and meta-analysis.

Tick-borne pathogens of the genus Anaplasma cause anaplasmosis in livestock and humans, impacting health and livelihoods, particularly in Africa. A comprehensive review on the epidemiology of Anaplasma species is important to guide further research and for implementation of control approaches. We reviewed observational studies concerning Anaplasma species amongst cattle in Africa. Peer-reviewed studies published in PubMed, Google Scholar, and Web of Science - from database inception to 2022 - were searched. The quality of individual studies was assessed using the Joanna Briggs Institute Critical Appraisal Tool and the pooled prevalences by diagnostic method were estimated using random-effects models. Heterogeneity across the studies was tested and quantified using the Cochran's Q statistic and the I2 statistic. Potential sources of heterogeneity were investigated by subgroup analysis. A total of 1117 records were retrieved and at the end of the screening, 149 records (155 studies) were eligible for this meta-analysis. The occurrence of Anaplasma species was reported in 31/54 countries in all regions. Seven recognised species (A. marginale, A. centrale, A. phagocytophilum, A. platys, A. capra, A. bovis, A. ovis) and nine uncharacterised genotypes (Anaplasma sp. Hadesa; Anaplasma sp. Saso; Anaplasma sp. Dedessa; Anaplasma sp. Mymensingh; Anaplasma sp. Lambwe-1; Candidatus Anaplasma africae; Anaplasma sp.; Candidatus Anaplasma boleense) were reported in African cattle. Anaplasma marginale was the most frequently reported (n=144/155 studies) and the most prevalent species (serology methods 56.1%, 45.9-66.1; direct detection methods 19.9%, 15.4-24.7), followed by A. centrale (n=26 studies) with a prevalence of 8.0% (95% CI: 4.8-11.9) and A. platys (n=19 studies) with prevalence of 9.7% (95% CI: 5.4-15.2). Anaplasma marginale, A. centrale and A. platys were reported in all Africa's regions, while A. ovis and A. capra were reported only in the northern and central regions. The uncharacterised Anaplasma taxa were mostly detected in the eastern and southern regions. Subgroup analysis showed that significant determinants for A. marginale exposure (serology) were geographical region (p=0.0219), and longitude (p=0.0336), while the technique employed influenced (p<0.0001) prevalence in direct detection approaches. Temperature was the only significant variable (p=0.0269) for A. centrale. These findings show that various Anaplasma species, including those that are zoonotic, circulate in African cattle. There is need for more genetic and genome data, especially for unrecognised species, to facilitate effective identification, improve livestock and minimise the health risk in human populations. Additional epidemiological data including pathogen occurrence, tick vectors and host range, as well as pathogenicity are essential.

Identification and distribution of Rhipicephalus microplus in selected high-cattle density districts in Uganda: signaling future demand for novel tick control approaches.

BACKGROUND: Rhipicephalus (Boophilus) microplus (Canestrini, 1888), the Asian blue tick, is a highly invasive and adaptable ectoparasite. This tick species has successfully established itself in most regions of the world, with movement of cattle being a major driver for its spread. In the recent past, R. microplus ticks have been reported in three districts of Uganda. Information on its spread and distribution are vital in deepening our understanding of the ecological scenarios that lead to tick persistence and in the formulation of control strategies. This is especially important in the cattle-dense districts. METHODS: We randomly collected tick specimens from 1,461cattle spread across seven cattle dense districts located in the Central, Karamoja and West Nile regions of Uganda from January to September 2020. The ticks were identified using standard morpho-taxonomic keys and the R. microplus tick species identities were confirmed by sequencing of the ITS2 region, 12S rRNA and 16S rRNA genes and phylogenetic analyses. RESULTS: Adult ticks (n = 13,019) were collected from 1,461 cattle. Seventeen tick species were identified based on morpho-taxonomic keys and the majority (47.4%; n=6184) of these were R. appendiculatus. In total, 257 R. microplus ticks were found infesting cattle in 18 study sites in the districts of Amudat, Kaabong, Napak (Karamoja region) and Arua (West Nile region). The identity of R. microplus was confirmed using molecular technics. No R. microplus tick was recorded in the districts of Lyantonde and Nakaseke (Central region). Arua district accounted for 82.1% (n=211) of the R. microplus ticks recorded followed by Napak district at 16.3% (n=42), while Amudat and Kaabong districts accounted for 1.5% (n=4). Rhipicephalus microplus and R. decoloratus co-existed in 6 of the 13 study sites in Arua district, while in another 6 study sites, no R. decoloratus was recorded. In the Karamoja region districts R. decoloratus co-existed with R.microplus. Of the total 618 ticks belonging to four species of the subgenus Boophilus recorded in this study, R. decoloratus accounted for 50.04% (n=334), followed by R. microplus at 41.58% (n=257), R. geigyi at 2.75% (n=17) and R. annulatus at 1.61% (n=10). In the districts of Amudat, Kaabong and Napak, R. decoloratus was more dominant (76.1%; n=179) of the three Rhipicephalus (Boophilus) tick species recorded, followed by R. microplus (19.5%; n=46) and R. geigyi (4.2%; n=10). Contrariwise, R. microplus was more dominant (84%; n=211) in Arua district followed by R. decoloratus (10.7%; n=27), R. annulatus (3.9%; n=10) and R. geigyi (1.1%; n=3). Phylogenetic analyses of the ITS2 region, 12S rRNA and 16S rRNA genes revealed subgrouping of the obtained sequences with the previously published R. microplus sequences from other parts of the world. CONCLUSION: Rhipicephalus microplus ticks were found infesting cattle in four districts of Uganda. The inability to find R. decoloratus, an indigenous tick, from six sites in the district of Arua is suggestive of its replacement by R. microplus. Rhipicephalus microplus negatively affects livestock production, and therefore, there is a need to determine its distribution and to deepen the understanding of the ecological factors that lead to its spread and persistence in an area.

Brucellosis Seropositivity Using Three Serological Tests and Associated Risk Factors in Abattoir Workers in Gauteng Province, South Africa.

Abattoir workers are liable to zoonotic infections from animals and animal products, primarily to diseases with asymptomatic and chronic clinical manifestations in animals, such as brucellosis. No published reports exist on the seroprevalence of brucellosis in abattoir workers in South Africa. Therefore, this cross-sectional study was conducted to estimate the occurrence and risk factors for Brucella exposure in abattoir workers in Gauteng Province. A total of 103 abattoir workers and managers from 6 abattoirs, where brucellosis-positive slaughtered cattle and sheep were previously detected, were interviewed and tested with serological assays using the Rose Bengal test (RBT), BrucellaCapt, and IgG-ELISA. A pre-tested questionnaire was administered to consenting respondents to obtain information on risk factors for brucellosis. Of the 103 respondents tested, the distribution of female and male workers was 16 (15.5%) and 87 (84.5%), respectively. The seroprevalence for exposure to brucellosis was 21/103 (20.4%, 95%CI: 13.1-29.5) using a combination of RBT, BrucellaCapt, or IgG-ELISA. For test-specific results, seroprevalences by RBT, BrucellaCapt, and IgG-ELISA were 13/103 (12.6%, 95%CI: 6.9-20.6), 9/103 (8.74%, 95%CI: 4.1-15.9), and 18/103 (17.5%, 95%CI: 10.7-26.2), respectively. Low-throughput abattoirs were identified as associated risks, as 29.3% of workers were seropositive compared with 12.7% of workers in high-throughput abattoirs, which highlights that direct contact at abattoirs poses higher risk to workers than indirect and direct contact outside abattoirs. This study confirms the occurrence of Brucella spp. antibodies among abattoir workers in South Africa, possibly due to occupational exposure to Brucella spp., and highlights the occupational hazard to workers. Furthermore, findings underscore that abattoir facilities can serve as points for active and passive surveillance for indicators of diseases of public health importance. We recommend periodic implementation of brucellosis testing of abattoir workers country-wide to establish baseline data for informing appropriate preventive practices and reducing the potential burden of infection rates among these high-risk workers.

Epidemiology of Echinococcus granulosus sensu lato in the Greater Horn of Africa: A systematic review.

BACKGROUND: Cystic echinococcosis (CE) is a neglected zoonotic disease that is caused by Echinococcus granulosus sensu lato (s.l.), the life cycle of which involves multiple hosts. We conducted a systematic review (SR) on E. granulosus s.l. in the Greater Horn of Africa (GHA), to provide a picture of its recent epidemiology across all hosts. METHODS: For this SR, conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement, five electronic databases, as well experts in the region were consulted to retrieve records published between 2000 and 2022, reporting the presence of E. granulosus s.l. infections in any natural host in the GHA (Djibouti, Eritrea, Ethiopia, Kenya, Sudan, Somalia, South Sudan, Tanzania and Uganda). PRINCIPAL FINDINGS: A total of 247 records were retained, describing the presence of E. granulosus s.l. throughout the GHA, except for Djibouti. Only few population surveys on human CE were conducted in the area, with the prevalence ranging between 0.3 and 11.3%. In animals, the reported prevalence ranged up to 61.6% in camels, 88.4% in cattle; 65.2% in goats, 9.9% in pigs, 67.8% in sheep and 94.5% in dogs. In addition, E. granulosus s.l. was also reported in wildlife. A total of five species were reported in the different hosts, namely E. granulosus sensu stricto (G1, G3, GOmo), E. canadensis (G6/7), E. ortleppi (G5), E. felidis, and E. equinus (G4). CONCLUSIONS: We confirm that E. granulosus s.l. is prevalent throughout the GHA. Nevertheless, despite our efforts to screen grey literature, an accurate assessment of the epidemiology in GHA remains challenging, due to the lack of combined host, in-depth risk factor and behavioural studies, as well as the wide diversity in subpopulations studied and diagnostic tools used. Interdisciplinary and transboundary partnerships would be essential for the design of effective control strategies, tuned to the GHA setting.

Polyphasic Characterization of Brucella spp. in Livestock Slaughtered from Abattoirs in Eastern Cape, South Africa.

In livestock, brucellosis is mainly an asymptomatic disease except when abortion occurs; therefore, two serological tests are used for diagnosis as no single test is suitable. Abattoir samples enable a combination of culture, molecular, and serological tests to detect brucellosis. This study assessed Brucella-specific PCR (ITS-PCR) to detect brucellosis and to conduct a molecular characterization of Brucella spp. isolated from PCR-positive livestock (n = 565) slaughtered at abattoirs and the appropriate sample tissue(s). ITS-PCR detected Brucella DNA in 33.6% of cattle, 14.5% of sheep, and 4.7% of pig tissues. Impure Brucella cultures from PCR-positive tissues were 43.6% (44/94) of cattle, 51.7% (15/29) of sheep, and 50% (2/4) of pigs with predominantly B. abortus identification with AMOS-PCR and low isolation of mixed B. abortus and B. melitensis in all species. In cattle, 33% of isolates were from lymph nodes, while in sheep 38.0% were from the liver and kidney and only from tonsils in pigs (2/4). Brucella infections identified with AMOS-PCR were present in seropositive and mainly seronegative (75.6-100%) livestock with the potential to cause brucellosis during pregnancy or breeding. This study demonstrated the value of the polyphasic approach, especially with chronic infections and the potential risk of these asymptomatic animals.

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

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

Molecular Characterization of Leptospira Species Detected in the Kidneys of Slaughtered Livestock in Abattoirs in Gauteng Province, South Africa.

Leptospira was investigated in kidneys (n = 305) from slaughtered livestock in the Gauteng Province abattoirs, South Africa, using a culture medium to isolate Leptospira, followed by the LipL32 qPCR to detect Leptospira DNA. The SecY gene region was amplified, sequenced, and analyzed for LipL32 qPCR-positive samples or Leptospira isolates. The overall frequency of isolation of Leptospira spp. was 3.9% (12/305), comprising 4.8% (9/186), 4.1% (3/74), and 0% (0/45) from cattle, pigs, and sheep, respectively (p > 0.05). However, with LipL32 qPCR, the overall frequency of Leptospira DNA was 27.5%, consisting of 26.9%, 20.3%, and 42.2% for cattle, pigs, and sheep, respectively (p = 0.03). Based on 22 SecY sequences, the phylogenetic tree identified the L. interrogans cluster with serovar Icterohaemorrhagiae and the L. borgpetersenii cluster with serovar Hardjo bovis strain Lely 607. This study is the first molecular characterization of Leptospira spp. from livestock in South Africa. The reference laboratory uses an eight-serovar microscopic agglutination test panel for leptospirosis diagnosis, of which L. borgpetersenii serovar Hardjo bovis is not part. Our data show that pathogenic L. interrogans and L. borgpetersenii are circulating in the livestock population. Diagnostic use of molecular methods will eliminate or reduce the under-reporting of leptospirosis in livestock, particularly sheep, in South Africa.

The knowledge, attitudes, and practices of smallholder cattle farmers concerning the epidemiology of bovine fasciolosis in the North West Province, South Africa.

Bovine fasciolosis has negative impacts on cattle production worldwide, more so on the African continent and especially in smallholder farming areas with limited level of awareness. A cross-sectional questionnaire-based survey was conducted to investigate the knowledge, attitudes, and practices concerning bovine fasciolosis among smallholder cattle farmers in the North West Province of South Africa. A total of 153 farmers were interviewed from three villages of the Moretele Local Municipality in Bojanala District. The majority of respondents were male (84%) farm owners (81%) that had low education levels (56% primary school or less) and employed extensive cattle management systems (84%). A large number of farms lacked infrastructure including calving pens (88%), restraining equipment (85%), and weight determination equipment (92%) while sourcing drinking water for cattle from rivers or dams (58%). No evaluated factors were significantly associated with a positive fasciolosis epidemiological knowledge score. However, education level (P = 0.046), some cattle breeds (P = 0.022), and management system (P < 0.001) of the smallholder farmers were associated with a positive practice score concerning bovine fasciolosis prevention. We therefore recommend that education programs be introduced that focus on the mode of transmission, risk factors, zoonotic importance, and practices associated with the prevention and control of bovine fasciolosis.

Genetic diversity in Babesia bovis from southern Africa and estimation of B. bovis infection levels in cattle using an optimised quantitative PCR assay.

Babesia bovis is a causal agent of bovine babesiosis, a disease which leads to mortality and morbidity and impacts the cattle industry worldwide. We amplified, cloned and sequenced the B. bovis merozoite surface antigen-2b (msa-2b) gene (∼940 bp) and the near full-length 18S rRNA gene (∼1600 bp) from cattle samples from South Africa and Mozambique to determine sequence variation between B. bovis parasites in the region. A TaqMan quantitative real-time PCR (qPCR) assay (18S rRNA gene) was optimised for the detection of B. bovis and estimation of parasitaemia in field samples from cattle from southern Africa. Phylogenetic analysis grouped the Msa-2b sequences in six clades and these were 59.7 to 99.6% identical to reference sequences. Sequence variation amongst B. bovis 18S rRNA sequences was found at 2 to 36 positions, and the sequences were 97 to 99% identical to published sequences. Mismatches between the B. bovis 18S rRNA sequences and a previously published qPCR forward primer (BoF) were observed; therefore, we developed a new forward primer (BoF2), and optimised the qPCR assay. Six 10-fold dilution series of B. bovis infected erythrocytes (2 × 108 to 2 × 103 infected red blood cells [iRBC]/ml) were analysed in triplicate in each of six separate qPCR runs, to determine the efficiency of the assay. The qPCR assay amplified the B. bovis 18S rRNA gene with 92.0 to 94.9% efficiency. The detection limit of the qPCR assay was approximately 6 iRBCs/µl. The performance of the optimised assay to diagnose B. bovis in field samples was assessed by testing DNA from 222 field samples of cattle from South Africa and Mozambique using three methods: the optimised qPCR assay, the reverse line blot (RLB) hybridisation assay, and the previously published qPCR assay. The detection rate of B. bovis using the optimised qPCR assay (31.1%, 69/222) was significantly higher (p<0.001) than both that using RLB (20.7%, 46/222) and the previously published qPCR assay (5.4%; 12/222). The B. bovis parasitaemia in samples from infected cattle ranged from 6 iRBCs/µl to 101,852 iRBCs/µl of blood. Our study revealed marked sequence variation between B. bovis parasites from southern Africa. The optimised qPCR assay will be useful in epidemiological studies and clinical diagnosis of B. bovis in southern Africa, and can be used to determine parasitaemia and potential carrier status in cattle populations, which is essential in the control of babesiosis.

Assessment of knowledge, attitudes and practices relating to brucellosis among cattle farmers, meat handlers and medical professionals in Namibia.

BACKGROUND: Brucellosis is a re-emerging zoonosis of significant socio-economic, animal and public health importance. It is principally a foodborne or occupation-associated infection of humans, whose effective control depends on maximum cooperation of high-risk populations. OBJECTIVES: The study assessed knowledge, attitudes and practices relating to brucellosis among cattle farmers (communal and commercial), meat handlers (abattoir and butchery workers) and medical professionals (nurses and doctors) in Namibia. METHODS: Between June 2019 and September 2020, self-administered questionnaires and questionnaire interviews were carried out in cattle farmers (n = 264), meat handlers (n = 143) and medical professionals (n = 124) in Namibia. RESULTS: Overall, 43.50% (231/531) of respondents were aware of brucellosis, with the highest awareness among medical professionals (73.39%, 91/124) and the least in meat handlers (13.99%, 20/143). Awareness of brucellosis was associated with tertiary education (p < 0.001) and the medical profession (p < 0.001). However, most medical professionals (98.39%, 122/124) did not consider brucellosis as a differential diagnosis in cases of persistent febrile illness. A proportion of communal (85.60%) and commercial (71.00%) farmers; abattoir workers (44.40%); butchers (53.50%); nurses (55.60%); and medical doctors (28.00%) consumed raw milk. CONCLUSIONS: The study identified the purchase of animals of unknown health status; assisting cow delivery; handling of aborted fetuses with no protective wear; consumption of raw milk, homemade cheese, cattle testes and undercooked livers, as risk factors for Brucella infection in cattle and humans. Thus, intensified risk communication, including public health education, is recommended, in particular, among meat handlers and communal farmers, to promote awareness and discourage risky practices.

Comparison of Three Diagnostic Methods to Detect the Occurrence of Fasciola Species in Communally Grazed Cattle in the North West Province, South 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.

Sex as a risk factor for occurrence and severity of infectious and parasitic diseases in dogs: Protocol for a systematic review.

Biological sex is an important risk factor for the occurrence and severity of infectious and parasitic diseases. Although various studies and reviews have described sex differences in infectious diseases of humans, wildlife and laboratory animals, there has been little focus on biological sex as a risk factor for infectious and parasitic diseases of domestic animals. We aim to identify and synthesise evidence in dogs for the hypothesis that biological sex and gonadectomy status are determinants of occurrence and severity of disease across taxa of pathogens. This systematic review follows the Preferred Reporting Items for Systematic reviews and Meta-analyses (PRISMA) guidelines. We will search Web of Science, Scopus and PubMed for peer-reviewed studies published in English from database inception through 2021. All study designs for infectious and parasitic diseases of dogs will be included. This review will include the outcomes prevalence or incidence of infection or disease; and severity of disease as measured by case-fatality, time to death or recovery, hospitalisation time, pathogen burden (e.g. viral load or parasitaemia) or relevant clinicopathological parameters. Two reviewers will jointly assess the first 500 records from all three databases. Subsequently, one reviewer will screen the remaining records, and then the second reviewer will verify all records excluded by the first reviewer. Full-texts of all included records will be retrieved and assessed for eligibility by the first review author, and then the second author will review those records excluded by the first author. The risk of bias in individual studies will be assessed using the Risk of Bias Assessment tool for Nonrandomized Studies. We will synthesise the information from the studies and present this as a narrative in the text. The findings will be presented by outcome type and also grouped by pathogen type. Evidence on sex-specific effects will expand our understanding of infectious disease pathogenesis and underlying mechanisms, and this may be of importance in implementation of disease control interventions.

Sex-differential non-specific effects of adjuvanted and non-adjuvanted rabies vaccines versus placebo on all-cause mortality in dogs (NERVE-Dog study): a study protocol for a randomized controlled trial with a nested case-control study.

BACKGROUND: It has been proposed that childhood vaccines in high-mortality populations may have substantial impacts on mortality rates that are not explained by the prevention of targeted diseases, nor conversely by typical expected adverse reactions to the vaccines, and that these non-specific effects (NSEs) are generally more pronounced in females. The existence of these effects, and any implications for the development of vaccines and the design of vaccination programs to enhance safety, remain controversial. One area of controversy is the reported association of non-live vaccines with increased female mortality. In a previous randomized controlled trial (RCT), we observed that non-live alum-adjuvanted animal rabies vaccine (ARV) was associated with increased female but not male mortality in young, free-roaming dogs. Conversely, non-live non-adjuvanted human rabies vaccine (NRV) has been associated with beneficial non-specific effects in children. Alum adjuvant has been shown to suppress Th1 responses to pathogens, leading us to hypothesize that alum-adjuvanted rabies vaccine in young dogs has a detrimental effect on female survival by modulating the immune response to infectious and/or parasitic diseases. In this paper, we present the protocol of a 3-arm RCT comparing the effect of alum-adjuvanted rabies vaccine, non-adjuvanted rabies vaccine and placebo on all-cause mortality in an owned, free-roaming dog population, with causal mediation analysis of the RCT and a nested case-control study to test this hypothesis. METHODS: Randomised controlled trial with a nested case-control study. DISCUSSION: We expect that, among the placebo group, males will have higher mortality caused by higher pathogen loads and more severe disease, as determined by haematological parameters and inflammatory biomarkers. Among females, we expect that there will be no difference in mortality between the NRV and placebo groups, but that the ARV group will have higher mortality, again mediated by higher pathogen loads and more severe disease. We anticipate that these changes are preceded by shifts in key serum cytokine concentrations towards an anti-inflammatory immune response in females. If confirmed, these results will provide a rational basis for mitigation of detrimental NSEs of non-live vaccines in high-mortality populations.

Body condition scores, fluke intensity, liver pathology, and carcass quality of different dairy cattle genotypes infected with Fasciola species at high throughput abattoirs in South Africa.

Milk is an essential commodity whose demand far exceeds supply. However, dairy animal productivity is constantly hampered by parasitic diseases such as fasciolosis, affecting milk production. Despite the negative impact of liver fluke on milk production, there is little information on liver fluke infection and associated abattoir losses (body weight, condition score, liver pathology, and carcass quality) in culled dairy cattle. This study aimed to determine body condition scores, fluke intensity, liver pathology, and carcass quality of different cattle genotypes infected with Fasciola species at three commercial abattoirs. A longitudinal study was conducted from September 2019 to October 2020 to determine body condition score, liver fluke intensity, liver pathology in 3065 dairy cattle slaughtered in CA1, CA2, and CA3, of the Eastern Cape Province South Africa. Liver fluke intensity significantly increased with cattle age (P < 0.0001). Cattle ≥ 7 years old (59.93 ± 6.42) and those 4 to 6 years old (49.78 ± 9.98) had higher infection than those 2 to 3 years old (27.55 ± 13.68). The liver fluke infection was significantly (P < 0.001) the highest when sampling was conducted in summer, followed by autumn and winter, and least for spring. The differences in carcass weights or body condition scores decreased by 0.99 units (P < 0.0001) or 0.97 units (P < 0.0001) respectively. Therefore, this study suggests that fluke infection could be responsible for considerable economic and production losses mainly due to condemnation and weight loss in dairy cattle. This study recommended a combination of holistic and grazing management to control infection rates in dairy herds.

Immunological Evidence of Variation in Exposure and Immune Response to Bacillus anthracis in Herbivores of Kruger and Etosha National Parks.

Exposure and immunity to generalist pathogens differ among host species and vary across spatial scales. Anthrax, caused by a multi-host bacterial pathogen, Bacillus anthracis, is enzootic in Kruger National Park (KNP), South Africa and Etosha National Park (ENP), Namibia. These parks share many of the same potential host species, yet the main anthrax host in one (greater kudu (Tragelaphus strepsiceros) in KNP and plains zebra (Equus quagga) in ENP) is only a minor host in the other. We investigated species and spatial patterns in anthrax mortalities, B. anthracis exposure, and the ability to neutralize the anthrax lethal toxin to determine if observed host mortality differences between locations could be attributed to population-level variation in pathogen exposure and/or immune response. Using serum collected from zebra and kudu in high and low incidence areas of each park (18- 20 samples/species/area), we estimated pathogen exposure from anti-protective antigen (PA) antibody response using enzyme-linked immunosorbent assay (ELISA) and lethal toxin neutralization with a toxin neutralization assay (TNA). Serological evidence of pathogen exposure followed mortality patterns within each system (kudus: 95% positive in KNP versus 40% in ENP; zebras: 83% positive in ENP versus 63% in KNP). Animals in the high-incidence area of KNP had higher anti-PA responses than those in the low-incidence area, but there were no significant differences in exposure by area within ENP. Toxin neutralizing ability was higher for host populations with lower exposure prevalence, i.e., higher in ENP kudus and KNP zebras than their conspecifics in the other park. These results indicate that host species differ in their exposure to and adaptive immunity against B. anthracis in the two parks. These patterns may be due to environmental differences such as vegetation, rainfall patterns, landscape or forage availability between these systems and their interplay with host behavior (foraging or other risky behaviors), resulting in differences in exposure frequency and dose, and hence immune response.

International interlaboratory validation of a nested PCR for molecular detection of Babesia bovis and Babesia bigemina, causative agents of bovine babesiosis.

Babesia bovis and B. bigemina are tick-transmitted parasites causing bovine babesiosis, characterized by significant morbidity and mortality leading to economic losses to the livestock industry in tropical and subtropical regions worldwide. Animals that recover from acute infection remain carriers with low parasitemia acting as a source of transmission, and often escape detection. An improved diagnosis of a B. bovis and/or B. bigemina infection of carrier animals is enabled by the availability of detection methods with high sensitivity. To this end, two nested PCR assays targeting the cytochrome b (cytb) genes of B. bovis and B. bigemina (cytb-nPCR), have been recently developed and an increased sensitivity with respect to reference protocols has been shown (Romero-Salas et al., 2016). In this study, the specificity against a panel of hemoparasites that potentially co-occur with B. bovis and B. bigemina was demonstrated to ensure applicability of the cytb-nPCR assays in a wide range of regions where bovine babesiosis is endemic. Furthermore, we compared both reported cytb-nPCR assays with reference nPCR and qPCR protocols for (i) their capability to detect carrier animals in the field, and (ii) their reproducibility when performed in different laboratories by independent operators. We show that, in a panel of bovine field samples (n = 100), the cytb-nPCR assays detected a considerably higher number of 25% B. bovis and 61% B. bigemina-positive animals compared to 7% and 20% B. bovis and 55% and 49% B. bigemina-positive animals when tested by reference nPCR and qPCR protocols, respectively. Cytb-nPCRs were also found superior in the detection of carrier animals when field samples from Africa were analyzed. In addition, both the B. bovis and B. bigemina cytb-nPCR assays were independently validated in a single blinded study in three laboratories. Importantly, no significant differences in the number/percentage of infected animals was observed using cytb-nPCR assays. In summary, the cytb-nPCR assays detected a considerably higher number of chronically infected B. bovis and B. bigemina carrier animals compared to reference nPCR and qPCR protocols, when applied in different epidemiological field situations. Furthermore, a high reproducibility between laboratories could be demonstrated.

Rickettsia africae infection rates and transovarial transmission in Amblyomma hebraeum ticks in Mnisi, Bushbuckridge, South Africa.

Rickettsia africae is a gram-negative bacterium, which causes African tick bite fever (ATBF) in humans. ATBF is a febrile disease mainly affecting travellers to southern Africa. This bacterium is known to be transmitted by Amblyomma hebraeum and Amblyomma variegatum ticks. In southern Africa, the principal vector is A. hebraeum. Febrile disease is a serious issue in the study area. There is a high prevalence of non-malaria illness caused by Rickettsia, so there is a need to have more knowledge on these species. Infection rates and transovarial transmission efficiency of R. africae in A. hebraeum ticks were investigated in a rural area of Mpumalanga province, South Africa. Adult and engorged A. hebraeum female ticks were collected from cattle. Larvae were collected by dragging a cloth at ground level using 100 steps, equivalent to an area of 100 m2. Tick identification was performed according to standard taxonomic keys using a microscope. Engorged ticks were incubated to oviposit and egg masses were collected. DNA was extracted from the ticks, larvae and egg masses, and screened for gltA and ompA genes, using quantitative real-time PCR and conventional PCR, respectively. Positive ompA amplicons were sequenced and phylogenetic analysis showed 99.8-100% identity with R. africae. Infection rates were 13.7 and 12.7% for adults and larvae, respectively. Transovarial transmission of R. africae in A. hebraeum from this study was 85.7%. The results provide a clear indication that people living in the study area and travellers that visit the area are at risk of contracting ATBF.

Prevalence, Risk Factors for Exposure, and Socio-Economic Impact of Peste Des Petits Ruminants in Karenga District, Karamoja Region, Uganda.

Peste des petits ruminants (PPR), a disease caused by small ruminant morbillivirus (SRM), is highly contagious with high morbidity and mortality. Controlling PPR requires a proper understanding of the epidemiological dynamics and impact of the disease in a range of geographical areas and management systems. Karenga district, located in the pastoral region of Karamoja in northeastern Uganda, and in the vicinity of Kidepo Valley National Park, is characterised by free cross-border (South Sudan and Kenya) livestock trade, communal grazing, and transhumance. This study was conducted from November through December 2020 to determine the seroprevalence of anti-SRM antibodies, the risk factors associated with the occurrence, and the socio-economic impact of PPR in Karenga. A total of 22 kraals were randomly selected from all administrative units, and 684 small ruminants (sheep = 115, goats = 569) were selected for serum collection using systematic random sampling. Exposure to SRM was determined using a competitive enzyme-linked immunosorbent assay. The overall true seroprevalence of SRM antibodies was high, 51.4 (95% confidence interval [CI] 45-52.6). Multivariate logistic regression for risk factors showed that seroprevalence varied significantly by location (26.8% to 87.8%, odds ratio (OR) ≤ 14.5). The odds of exposure to SRM were higher in sheep (73.9%) than in goats (43.8%) (OR = 1.7, p = 0.08), and seropositivity was higher in animals greater than two years old (65.5%; OR = 11.1, p < 0.001), or those one to two years old (24.7%; OR = 1.6, p = 0.2), compared to small ruminants less than one year old (16.1%). Using participatory epidemiology approaches (semi-structured interviews, clinical examinations, pairwise ranking, proportional piling, impact matrix scoring) with 15 key informants and 22 focus groups of pastoralists, PPR was the second most important small ruminant disease: relative morbidity 14%, relative mortality 9%, and case fatality rate 78%, and impacted productivity mainly in terms of treatment costs, mortality, marketability, and conflicts. These findings provide evidence to support the implementation of disease surveillance and control strategies to mitigate the impact of PPR in Karamoja and other pastoral areas in eastern Africa.

Molecular detection and characterisation of protozoan and rickettsial pathogens in ticks from cattle in the pastoral area of Karamoja, Uganda.

Ticks and tick-borne diseases (TBDs) significantly affect cattle production and the livelihoods of communities in pastoralist areas. Data on protozoan and rickettsial pathogens in ticks infesting cattle in Uganda is scanty; while it is an indicator of the likelihood of disease transmission and occurrence. A cross-sectional study was conducted amongst cattle in the Karamoja Region, northeastern Uganda, from July through September 2017, to determine the tick species diversity, identify protozoan and rickettsial pathogens in the ticks, and characterise pathogenic species by sequence and phylogenetic analyses. About 50 % of the ticks detected from each predilection site on each animal were collected from 100 purposively-selected cattle from 20 randomly-selected herds. Twelve tick species belonging to the genera Amblyomma, Rhipicephalus and Hyalomma were identified, the most abundant being Amblyomma lepidum (93.9 %), followed by Amblyomma variegatum (2.0 %) and Rhipicephalus evertsi evertsi (1.0 %). Tick species that have not been reported in recent studies amongst cattle in Uganda were found, namely Rhipicephalus pravus, Rhipicephalus praetextatus and Rhipicephalus turanicus. The ticks were grouped into 40 pools, by species and location, and the reverse line blot (RLB) hybridisation assay was used to detect pathogens from the ticks. The most frequently detected tick-borne parasites were Theileria mutans, Theileria velifera and Theileria parva, each observed in 25 % (10/40) of the tick pools. Tick-borne pathogens, namely Babesia rossi, Babesia microti and Theileria sp. (sable) that are not common to, or not known to infect, cattle were identified from ticks. The gene encoding Ehrlichia ruminantium pCS20 region, the Ehrlichia and Anaplasma 16S rRNA gene, and T. parva p67 sporozoite antigen gene were amplified, cloned and sequenced. Seven novel E. ruminantium pCS20 variants were identified, and these grouped into two separate clusters with sequences from other parts of Africa and Asia. The T. parva p67 sequences were of the allele type 1, and parasites possessing this allele type are commonly associated with East Coast fever in eastern Africa. Analysis of the Ehrlichia and Anaplasma 16S rRNA gene sequences showed that they were closely related to Rickettsia africae and to a new Ehrlichia species variant recently found in China. Our R. africae 16S rRNA sequences grouped with R. africae isolates from Nigeria, Egypt and Benin. The information on tick species diversity and pathogens in the various tick species provides an indicator of potential transmission amongst cattle populations, and to humans, and can be useful to estimate disease risk and in control strategies.

Ticks and tick-borne pathogens infecting livestock and dogs in Tchicala-Tcholoanga, Huambo Province, Angola.

The diversity of ticks and tick-borne pathogens (TBPs) infesting domestic animals in Tchicala-Tcholoanga, Angola, in 2016 was investigated. Seventeen tick species were recorded, Amblyomma pomposum being the most abundant on cattle (40%), goats (38%) and sheep (35%); Rhipicephalus turanicus was the most abundant on dogs (46%). This study presents new records of Haemaphysalis paraleachi, R. compositus, R. kochi and R. sulcatus in Angola, the first georeferenced population of Ha. leachi in southern Africa and the second record of R. microplus in Angola. Using the reverse line blot (RLB) hybridisation assay, fifteen TBP species were detected in blood samples from cattle (n = 88), goats (n = 82), sheep (n = 85) and dogs (n = 85). F The most frequently detected species were Theileria velifera in cattle (78%), Theileria ovis in sheep (80%) and Babesia vogeli in dogs (35%). Species-specific quantitative PCR assays detected Babesia bigemina in 43% (35/80) of blood samples of cattle, while E. ruminantium was detected in 4% (3/70) of blood samples and in 7% of A. pomposum ticks. Anaplasma platys was detected from cattle (18%) and sheep (6%) during RLB analysis. These findings constitute pioneering research in Angola.