An investigation into Toxoplasma gondii at the human-livestock-wildlife interface, South Africa.

Interface areas shared by humans, domestic and wild animals may serve as high transmission contexts for Toxoplasma gondii. However, knowledge about the epidemiology of T. gondii in such areas is currently limited. The present study assessed the seroprevalence of T. gondii in different hosts from Mpumalanga, South Africa. Furthermore, we investigated the local knowledge and related practices about T. gondii by conducting a questionnaire study in the community. Blood samples were obtained and analysed for T. gondii antibodies using a commercial multispecies latex agglutination kit. The seroprevalence detected in humans (n = 160; patients showing signs of acute febrile illness), cats (n = 9), chickens (n = 336) and goats (n = 358) was 8.8%, 0.0%, 4.2% and 11.2%, respectively. Seroprevalence in impalas (n = 97), kudus (n = 55), wild dogs (n = 54), wildebeests (n = 43), warthogs (n = 97) and zebras (n = 68) was calculated at 5.2%, 7.3%, 100.0%, 20.9%, 13.4% and 9.1%, respectively. The questionnaire revealed that 63.0% of household owners were subsistence farmers, and 35.9% were pet owners. A high level of female participation was found (75.3%) when compared to male participation (24.7%). The results show a low circulation of T. gondii in the domestic cycle and suggest the presence of possible bridges between the wildlife cycle and the surrounding domestic cycle.Contribution: The study contributes to identifying transmission patterns and risk factors of T. gondii within human and animal populations. This topic fits within the scope of the journal presenting original research in veterinary science, with the focus on wild and domestic populations on the African continent on a topic of universal importance.

Genome-wide diversity, population structure and signatures of inbreeding in the African buffalo in Mozambique.

The African buffalo, Syncerus caffer, is a key species in African ecosystems. Like other large herbivores, it plays a fundamental role in its habitat acting as an ecosystem engineer. Over the last few centuries, African buffalo populations have declined because of range contraction and demographic decline caused by direct or indirect human activities. In Mozambique, historically home to large buffalo herds, the combined effect of colonialism and subsequent civil wars has created a critical situation that urgently needs to be addressed. In this study, we focused on the analysis of genetic diversity of Syncerus caffer caffer populations from six areas of Mozambique. Using genome-wide SNPs obtained from ddRAD sequencing, we examined the population structure across the country, estimated gene flow between areas under conservation management, including national reserves, and assessed the inbreeding coefficients. Our results indicate that all studied populations of Syncerus caffer caffer are genetically depauperate, with a high level of inbreeding. Moreover, buffaloes in Mozambique present a significant population differentiation between southern and central areas. We found an unexpected genotype in the Gorongosa National Park, where buffaloes experienced a dramatic population size reduction, that shares a common ancestry with southern populations of Catuane and Namaacha. This could suggest the past occurrence of a connection between southern and central Mozambique and that the observed population structuring could reflect recent events of anthropogenic origin. All the populations analysed showed high levels of homozygosity, likely due to extensive inbreeding over the last few decades, which could have increased the frequency of recessive deleterious alleles. Improving the resilience of Syncerus caffer caffer in Mozambique is essential for preserving the ecosystem integrity. The most viable approach appears to be facilitating translocations and re-establishing connectivity between isolated herds. However, our results also highlight the importance of assessing intraspecific genetic diversity when considering interventions aimed at enhancing population viability such as selecting suitable source populations.

Pulmonary Telerehabilitation for People With Chronic Obstructive Pulmonary Disease in Brazil: A Mixed-Methods Feasibility Study.

This study assessed the feasibility of pulmonary telerehabilitation‧s (PTR) acceptability, implementation, practicality, and adaptation for people with Chronic Obstructive Pulmonary Disease (COPD) in Brazil. It also explored associations with clinical and socioeconomic features of Brazilians with COPD. This mixed-method study included thirty-one participants with COPD (age 62±10 years; FEV1= 72±14% predicted). Most participants (74.2%) reported good PTR session acceptability on the System Usability Scale and scores of 4.6±0.3 and 4.5±0.6 on a 1-5 Likert-type scale of implementation and practicality, respectively. Participants suggested adaptations for better comfort on the exercise bike and varying exercise modalities. PTR acceptability was associated with participants' younger age (rs=-0.57, p<0.01) and higher education (rs=0.51, p<0.01). PTR is feasible for people with COPD in Brazil regarding acceptability, implementation, practicality, and adaptation. Younger age and higher educational level are associated with greater PTR acceptability.

Characterisation of South African field Ehrlichia ruminantium using multilocus sequence typing.

Heartwater, one of the major tick-borne diseases of some domestic and wild ruminants in Africa, is caused by Ehrlichia ruminantium. The genetic diversity of E. ruminantium isolates renders the available vaccine ineffective against certain virulent isolates. To better understand the E. ruminantium genotypes in South Africa, a total of 1004 Amblyomma hebraeum tick deoxyribonucleic acid (DNA) samples from cattle in three South African provinces were tested by pCS20 Sol1 real-time polymerase chain reaction (qPCR) and characterised by multilocus sequence typing (MLST) using five housekeeping genes. Out of 1004 samples tested, 222 (22%) were positive for E. ruminantium. The occurrence of E. ruminantium in Mpumalanga, KwaZulu-Natal and Limpopo provinces was 19%, 22% and 27%, respectively. The E. ruminantium positive samples were screened for housekeeping genes and sequenced. Phylogenetic analysis revealed three main lineages: clade 1 made up of worldwide isolates (eastern, southern Africa, and Caribbean isolates), clade 2 comprised only West African isolates and clade 3 consisted of Omatjenne, Kümm2 and Riverside. Some study sample sequences were not identical to any of the reference isolates. However, they could all be grouped into the worldwide clade. Genetic variation in the sequenced regions was observed in the form of single nucleotide polymorphisms (SNPs). Using MLST to characterise E. ruminantium field isolates allowed the South African genotypes to be clearly distinguished from the distinct West African isolates.Contribution: Characterisation of E. ruminantium field isolates is important for the control of heartwater and contributes to preliminary knowledge required for the development of a more practical vaccine against heartwater.

The Brazilian version of the telehealth usability questionnaire (telehealth usability questionnaire Brazil): translation, cross-cultural adaptation, and psychometric properties.

OBJECTIVE: The objectives of this study were to translate and cross-culturally adapt the telehealth usability questionnaire into Brazilian Portuguese and to evaluate its psychometric properties. METHODS: This was a methodological validation study carried out in two phases. In phase 1, the telehealth usability questionnaire was cross-culturally adapted with 10 participants comprising the expert committee members, including 5 healthcare professionals with theoretical and practical knowledge of telehealth, 1 methodologist, and 4 translators. This phase was performed at Universidade Federal de Juiz de Fora Physiotherapy Clinic School. In phase 2, the psychometric properties of telehealth usability questionnaire Brazil were analyzed. This phase included in-person assessments at Márcio Cunha Hospital, Minas Gerais. The recruitment period for both phases was from April 2020 to February 2021. Content validity, reliability, internal consistency, and criterion validity were analyzed. The criterion validity was evaluated using correlation with a validated instrument: the system usability scale. RESULTS: The telehealth usability questionnaire was adequately translated and cross-culturally adapted. The telehealth usability questionnaire Brazil presented an excellent content validity index of 0.96 with percentages of understanding higher than 90%. The telehealth usability questionnaire Brazil demonstrated great internal consistency (α=0.94 and ω=0.94), excellent intra-rater reliability (intraclass correlation coefficient=0.85, 95%CI 0.75-0.91), no difference between the test and retest [T (0.425), p>0.673], and no proportional bias (p=0.205). There was a moderate correlation between telehealth usability questionnaire Brazil and the system usability scale (r=0.52, p<0.0001). CONCLUSION: The telehealth usability questionnaire was adequately translated and cross-culturally adapted into Brazilian Portuguese and showed adequate psychometric properties for use in telehealth clinical practice and research in Brazilian-Portuguese-speaking individuals.

Development and characterization of microsatellite markers for the tsetse species Glossina brevipalpis and preliminary population genetics analyses.

Tsetse flies, the vectors of African trypanosomes are of key medical and economic importance and one of the constraints for the development of Africa. Tsetse fly control is one of the most effective and sustainable strategies used for controlling the disease. Knowledge about population structure and level of gene flow between neighbouring populations of the target vector is of high importance to develop appropriate strategies for implementing effective management programmes. Microsatellites are commonly used to identify population structure and assess dispersal of the target populations and have been developed for several tsetse species but were lacking for Glossina brevipalpis. In this study, we screened the genome of G. brevipalpis to search for suitable microsatellite markers and nine were found to be efficient enough to distinguish between different tsetse populations. The availability of these novel microsatellite loci will help to better understand the population biology of G. brevipalpis and to assess the level of gene flow between different populations. Such information will help with the development of appropriate strategies to implement the sterile insect technique (SIT) in the framework of an area-wide integrated pest management (AW-IPM) approach to manage tsetse populations and ultimately address the trypanosomoses problem in these targeted areas.

Title: Développement et caractérisation de marqueurs microsatellites pour l'espèce de mouche tsé-tsé Glossina brevipalpis et analyses génétiques préliminaires des populations. Abstract: Les mouches tsé-tsé, vecteurs des trypanosomes africains, sont d'une importance médicale et économique majeure et l'une des contraintes pour le développement de l'Afrique. La lutte contre la mouche tsé-tsé est l'une des stratégies les plus efficaces et durables utilisées pour contrôler la maladie. La connaissance de la structure de la population et du niveau de flux de gènes entre les populations voisines du vecteur cible est d'une grande importance pour développer des stratégies appropriées pour la mise en œuvre de programmes de gestion efficaces. Les microsatellites sont couramment utilisés pour identifier la structure de la population et évaluer la dispersion des populations cibles et ont été développés pour plusieurs espèces de glossines mais manquaient pour Glossina brevipalpis. Dans cette étude, nous avons criblé le génome de G. brevipalpis pour rechercher des marqueurs microsatellites appropriés et neuf ont été trouvés suffisamment efficaces pour faire la distinction entre différentes populations de glossines. La disponibilité de ces nouveaux locus microsatellites aidera à mieux comprendre la biologie des populations de G. brevipalpis et à évaluer le niveau de flux de gènes entre différentes populations. Ces informations aideront à l'élaboration de stratégies appropriées pour mettre en œuvre la technique de l'insecte stérile dans le cadre d'une approche de lutte antiparasitaire intégrée à l'échelle de la zone pour gérer les populations de glossines et, en fin de compte, résoudre le problème des trypanosomoses dans les zones concernées.

Sympatry of Amblyomma eburneum and Amblyomma variegatum on African buffaloes and prevalence of pathogens in ticks.

The Amblyomma genus is represented on the African continent by 24 species, out of which 17 are known to occur in different ecological niches of southern Africa. Amblyomma, known for their aggressive hunting behaviour and aptitude as pathogen vectors, are of main concern to travellers, mainly in rural and conservation areas of Africa. In this study, we highlight the overlapping distribution of Amblyomma eburneum and Amblyomma variegatum found on African buffaloes (Syncerus caffer) at Coutada 11, Central Mozambique. In total, 1,039 Amblyomma ticks were collected and morphologically identified using taxonomic keys, and genomic DNA was extracted. They were subjected to reverse line blotting for pathogen identification followed by molecular analysis (COI sequencing) of both tick species. Pathogens such as Ehrlichia ruminantium, Anaplasma centrale, Theileria sp., Babesia sp. and Rickettsia africae were detected, of which R. africae is zoonotic. Ehrlichia ruminantium, R. africae, Theileria mutans and Theileria velifera are well-established pathogens transmitted by Amblyomma ticks; however, Anaplasma spp. and Babesia spp. are not, suggesting residual parasite DNA in the bloodmeal. Little is mentioned in the literature about A. eburneum, including its role as a vector and reservoir for pathogens. In Mozambique A. eburneum is currently restricted to wildlife but the spread of the tick may be observed given the climate change that is occurring. The infection rates for the pathogens in both Amblyomma tick species were lower than expected, but this may be due to the low host density in the forest niche and the innate immunity of these hosts. With the propensity of ticks of the Amblyomma genus to form parapatric distributions, the mechanisms that allows for the overlapping distribution of these two Amblyomma species while maintaining tick species identity is of great interest.

Seroprevalence and associated risk factors of bovine neosporosis in the Khomas region of Namibia.

Neospora caninum is a coccidian parasite that occurs worldwide and is one of the most important causes of abortion, especially in cattle. However, no studies have been performed in Namibia to determine the N. caninum status in livestock. Therefore, this study aimed to determine the seroprevalence of N. caninum in cattle and the associated risk factors in the Khomas region of Namibia. A total of 736 sera were collected from cows in 32 farming establishments. These comprised 698 beef and 38 dairy cattle sera and were tested using a commercial enzyme-linked immunosorbent assay (ELISA) kit. Questionnaires were concurrently administered to determine possible risk factors associated with N. caninum seropositivity. A total of 42 sera were positive (all beef), giving an animal-level seroprevalence rate of 5.7%. Eight of the 32 establishments had at least one positive animal, giving a herd-level seroprevalence of 25%. There was no significant association between seropositivity and the presence of dogs, jackals, history of abortions, farm size, number of cattle or average annual rainfall. The establishments with moderate to high numbers of Feliformia were 9.8 times more likely to be seropositive to N. caninum than those with none to low levels of the former (p = 0.0245). The authors concluded that the seroprevalence level of N. caninum in the Khomas region was relatively low compared with other parts of the world and that the role of Feliformia in the epidemiology of bovine neosporosis needed to be further investigated.Contribution: Serological evidence of bovine neosporosis and the associated risk factors are reported in Namibia for the first time. This study contributes to the scientific body of knowledge on N. caninum in Africa, which is currently limited.

Extensive field evidence for the release of HONO from the photolysis of nitrate aerosols.

Particulate nitrate ([Formula: see text]) has long been considered a permanent sink for NOx (NO and NO2), removing a gaseous pollutant that is central to air quality and that influences the global self-cleansing capacity of the atmosphere. Evidence is emerging that photolysis of [Formula: see text] can recycle HONO and NOx back to the gas phase with potentially important implications for tropospheric ozone and OH budgets; however, there are substantial discrepancies in "renoxification" photolysis rate constants. Using aircraft and ground-based HONO observations in the remote Atlantic troposphere, we show evidence for renoxification occurring on mixed marine aerosols with an efficiency that increases with relative humidity and decreases with the concentration of [Formula: see text], thus largely reconciling the very large discrepancies in renoxification photolysis rate constants found across multiple laboratory and field studies. Active release of HONO from aerosol has important implications for atmospheric oxidants such as OH and O3 in both polluted and clean environments.

The Brazilian version of the telehealth usability questionnaire (telehealth usability questionnaire Brazil): translation, cross-cultural adaptation, and psychometric properties

SUMMARY OBJECTIVE: The objectives of this study were to translate and cross-culturally adapt the telehealth usability questionnaire into Brazilian Portuguese and to evaluate its psychometric properties. METHODS: This was a methodological validation study carried out in two phases. In phase 1, the telehealth usability questionnaire was cross-culturally adapted with 10 participants comprising the expert committee members, including 5 healthcare professionals with theoretical and practical knowledge of telehealth, 1 methodologist, and 4 translators. This phase was performed at Universidade Federal de Juiz de Fora Physiotherapy Clinic School. In phase 2, the psychometric properties of telehealth usability questionnaire Brazil were analyzed. This phase included in-person assessments at Márcio Cunha Hospital, Minas Gerais. The recruitment period for both phases was from April 2020 to February 2021. Content validity, reliability, internal consistency, and criterion validity were analyzed. The criterion validity was evaluated using correlation with a validated instrument: the system usability scale. RESULTS: The telehealth usability questionnaire was adequately translated and cross-culturally adapted. The telehealth usability questionnaire Brazil presented an excellent content validity index of 0.96 with percentages of understanding higher than 90%. The telehealth usability questionnaire Brazil demonstrated great internal consistency (α=0.94 and ω=0.94), excellent intra-rater reliability (intraclass correlation coefficient=0.85, 95%CI 0.75-0.91), no difference between the test and retest [T (0.425), p>0.673], and no proportional bias (p=0.205). There was a moderate correlation between telehealth usability questionnaire Brazil and the system usability scale (r=0.52, p<0.0001). CONCLUSION: The telehealth usability questionnaire was adequately translated and cross-culturally adapted into Brazilian Portuguese and showed adequate psychometric properties for use in telehealth clinical practice and research in Brazilian-Portuguese-speaking individuals.

Evaluation of knowledge, attitudes and practices regarding neosporosis and toxoplasmosis among farmers and animal health practitioners in Namibia.

This study assessed the knowledge, attitudes and practices of livestock farmers in Namibia's Khomas region and animal health practitioners (veterinarians and animal health technicians) in the whole country concerning neosporosis and toxoplasmosis. Structured questionnaires were used, and a total of 63 farmers and 51 animal health practitioners responded out of an estimated 560 farmers in the Khomas region and 300 veterinarians and veterinary technicians in the country. Only 15.9% of the livestock farmers (n = 63) had heard about neosporosis or toxoplasmosis or knew how animals get infected (p < 0.0001). Only 5% of the farmers knew the risks associated with keeping dogs and cats concerning neosporosis and toxoplasmosis, respectively (p < 0.0001). None of the 51 animal health practitioners routinely requested Neospora caninum or Toxoplasma gondii laboratory tests in cases of cattle, sheep or goat abortions. Although all animal health practitioners indicated they routinely interacted with livestock farmers, none regularly discussed neosporosis or toxoplasmosis. Only 3.9% of animal health practitioners (n = 51) indicated that they had ever discussed either neosporosis or toxoplasmosis at a farmers' gathering (p < 0.0001), and only 21.6% had talked to at least one cat owner about toxoplasmosis in the previous 12 months (p < 0.0001). The authors concluded that farmers in the Khomas region were generally unaware of neosporosis and toxoplasmosis but could change their attitudes and practices if educated. The animal health practitioners lacked a deeper understanding and appreciation of the two diseases, which is required to cultivate enough confidence to educate farmers. Sharing this research and other relevant information on the two diseases at farmers' meetings, veterinary congresses, journals and newsletters could help educate farmers and animal health practitioners. Such platforms are likely to succeed because both these groups use these forums to get new information.

Body fat estimated by equations based on anthropometric parameters correlates with bioelectrical impedance in patients undergoing bariatric surgery / Gordura corporal estimada por equações baseadas em parâmetros antropométricos correlaciona-se com a bioimpedância elétrica em pacientes submetidos à cirurgia bariátrica

Introduction: predictive equations to estimate body fat based on simple anthropometric parameters are easy to use in the clinical practice.Objective: to evaluate the relationship between predictive equations based on anthropometric parameters and bioelectrical impedance to estimate body fat in individuals undergoing bariatric surgery.Methods: a prospective and longitudinal study carried out with individuals undergoing bariatric surgery. Body weight, body mass index, waist circumference and body fat percentage estimated by anthropometric parameters and by impedance were evaluated at three moments, one month before, two and six months after surgery. Data were analyzed by one-way ANOVA for repeated measures with Holm-Sidak ́s post hoc or Friedman test with Tukey ́s post hoc, and Pearson or Spearman correlations, according to data distribution. Significance level adopted 5%.Results: twenty-five subjects composed the final sample. All anthropometric parameters reduced significantly over time (p<0.001). Except for Lean et al equation before surgery, the body fat percentage estimated by other formulas showed a strong correlation with impedance in all moments, with the highest correlation strength observed in Gómez-Ambrosi et al. equation.Conclusion: in the present study, the equations used showed a good correlation with bioelectrical impedance, and the Gómez-Ambrosi et al. equation as a better option to the use of bioimpedance to assess changes in body fat percentage of patients undergoing bariatric surgery for the treatment of severe obesity.

Introdução: Objetivos: analisar o consumo alimentar e os fatores associados ao estado nutricional de crianças menores de dois anos de vida.Método: Estudo de corte transversal realizado com uma amostra de 344 lactentes menores de dois anos de idade e suas respectivas mães, acompanhadas em Unidades de Saúde da Família. As variáveis sociodemográficas, antropométrica das mães e dos lactentes e o consumo alimentar dessas crianças foram avaliadas por meio de questionário estruturado. A força de associação entre as variáveis dependente e as independentes foi avaliada pelo odds ratio, tanto na análise univariada quanto na múltipla, com nível de significância de 5%.Resultados: A prevalência do estado nutricional inadequado foi de 38,08%. Observou que 29,09% das crianças menores de seis meses de idade não chegaram a receber leite materno de forma exclusiva ou o tempo de oferta foi inferior a 30 dias. Notou-se o consumo de alimentos ultraprocessados, principalmente, do suco industrializado no último mês [OR:1,96, IC 95%: 1,05-3,65], baixa ingestão de frutas e o hábito de comer assistindo televisão nos lactentes maiores de seis meses. Após ajuste para variáveis de confusão, permaneceram associadas ao estado nutricional: anemia gestacional [OR: 3,58 IC: 1,46-8,77] e trabalho materno [OR, 0,38, IC 95%: 0,18-0,80].Conclusão: A presença do estado nutricional inadequado, caracterizado pelo baixo ou excesso de peso, associou-se ao fato de a mãe trabalhar e à anemia gestacional. Ademais, constatou-se a participação precoce de alimentos ultraprocessados na alimentação das crianças menores de 24 meses de idade, substituindo alimentos considerados naturais e saudáveis, evidenciando assim práticas alimentares inadequadas frente às recomendações para a faixa etária.

Toxoplasma gondii in African Wildlife: A Systematic Review.

Toxoplasma gondii (T. gondii) is a protozoan parasite, which infects a wide variety of mammals and bird species globally. In large parts of the world, this parasite is relatively well documented in wildlife species, however, this topic is poorly documented in Africa. The current review systematically explores the presence and distribution of T. gondii in African wildlife species through a key word search in PubMed, Web of Science and CAB Direct. A total of 66 records were identified and included in the qualitative analysis, of which 19 records were retained for the quantitative synthesis. The presence of T. gondii was reported in a wide range of wildlife species, found in twelve countries, spread over the African continent. The retained records report a prevalence range of 6-100% in herbivores, 8-100% in omnivores and 14-100% in carnivores. In wild felines (cheetahs, leopards, and lions) a prevalence range of 33-100% was found. Reports from South Africa, and on the presence of T. gondii in lion were most common. Overall, the results indicate the scarcity of information on T. gondii in Africa and its circulation in wildlife. The lack of knowledge on the parasite in Africa, especially in areas at the human-livestock-wildlife interface, prevents us from understanding how prevalent it is on the continent, what strains are circulating in wildlife and what the most common routes of transmission are in the different habitats in Africa.

Pseudomonas boanensis sp. nov., a bacterium isolated from river water used for household purposes in Boane District, Mozambique.

A Gram-negative rod with a single polar flagellum was isolated from a freshwater reservoir used for household purposes in Boane District, near Maputo, Mozambique, and designated as strain DB1T. Growth was observed at 30-42 °C (optimum, 30-37 °C) and with 0.5-1.5 % NaCl. Whole-genome-, rpoD- and 16S rRNA-based phylogenies revealed this isolate to be distant from other Pseudomonas species with Pseudomonas resinovorans, Pseudomonas furukawaii and Pseudomonas lalkuanensis being the closest relatives. Phenotypic analyses of strain DB1T showed marked differences with respect to type strains P. resinovorans CCUG 2473T, P. lalkuanensis CCUG 73691T, P. furukawaii CCUG 75672T and Pseudomonas otiditis CCUG 55592T. Taken together, our results indicate that strain DB1T is a representative of a novel species within the genus Pseudomonas for which the name Pseudomonas boanensis is proposed. The type strain is DB1T (=CCUG 62977T=CECT 30359T).

Prevalence of Trypanosoma and Sodalis in wild populations of tsetse flies and their impact on sterile insect technique programmes for tsetse eradication.

The sterile insect technique (SIT) is an environment friendly and sustainable method to manage insect pests of economic importance through successive releases of sterile irradiated males of the targeted species to a defined area. A mating of a sterile male with a virgin wild female will result in no offspring, and ultimately lead to the suppression or eradication of the targeted population. Tsetse flies, vectors of African Trypanosoma, have a highly regulated and defined microbial fauna composed of three bacterial symbionts that may have a role to play in the establishment of Trypanosoma infections in the flies and hence, may influence the vectorial competence of the released sterile males. Sodalis bacteria seem to interact with Trypanosoma infection in tsetse flies. Field-caught tsetse flies of ten different taxa and from 15 countries were screened using PCR to detect the presence of Sodalis and Trypanosoma species and analyse their interaction. The results indicate that the prevalence of Sodalis and Trypanosoma varied with country and tsetse species. Trypanosome prevalence was higher in east, central and southern African countries than in west African countries. Tsetse fly infection rates with Trypanosoma vivax and T. brucei sspp were higher in west African countries, whereas tsetse infection with T. congolense and T. simiae, T. simiae (tsavo) and T. godfreyi were higher in east, central and south African countries. Sodalis prevalence was high in Glossina morsitans morsitans and G. pallidipes but absent in G. tachinoides. Double and triple infections with Trypanosoma taxa and coinfection of Sodalis and Trypanosoma were rarely observed but it occurs in some taxa and locations. A significant Chi square value (< 0.05) seems to suggest that Sodalis and Trypanosoma infection correlate in G. palpalis gambiensis, G. pallidipes and G. medicorum. Trypanosoma infection seemed significantly associated with an increased density of Sodalis in wild G. m. morsitans and G. pallidipes flies, however, there was no significant impact of Sodalis infection on trypanosome density.

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.

Development of Cathepsin L-like Real-Time PCR Assays for the Detection of African Animal Trypanosomosis (AAT) in South Africa.

African animal trypanosomosis (AAT), is an infectious parasitic disease of wildlife and livestock caused by multiple species and strains of Trypanosoma. In South Africa, it is restricted to northern KwaZulu-Natal (NKZN) and caused by Trypanosoma congolense and Trypanosoma vivax. A cross-sectional study was done to determine AAT prevalence in 384 goat samples and identify trypanosome species circulating in 60 cattle at dip tanks that are on the interface with the Hluhluwe-uMfolozi game reserve in NKZN. Both cattle and goat samples were analyzed using the buffy coat technique (BCT) and a polymerase chain reaction (PCR) assay targeting the internal transcribed spacer 1 (ITS) region. Cattle samples were further analyzed using an ITS quantitative real-time PCR (qPCR) assays designed for the detection of T. congolense, T. vivax, and T. brucei. None of the goat samples tested positive for Trypanosoma infections. The ITS qPCR assay detected Trypanosoma DNA in 30% of the cattle samples, while only 8.3% were positive with the ITS PCR and 11.7% were positive using BCT. Quantitative real-time PCR assays were designed to amplify a 98 bp, 137 bp, and 116 bp fragment of the cathepsin L-like (CATL) gene from T. brucei, T. theileri, and T. congolense, respectively. Each assay was shown to be efficient (>94%) and specific (109 to 102/101 copies/reaction) in the detection of Trypanosoma species. The CATL qPCR assays detected T. congolense and T. theileri infections in 33.3% of the cattle samples. The CATL qPCR assays also detected T. congolense infections in goats (23.1%) that were neither detected by BCT nor the ITS PCR. The CATL qPCR assays provide an additional, sensitive, and specific tool for Trypanosoma diagnostics. The presence of trypanosomes in goats suggests they might be potential reservoirs of infections to other livestock.

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

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