Limited impact of vector control on the population genetic structure of Glossina fuscipes fuscipes from the sleeping sickness focus of Maro, Chad.

Tsetse flies (genus Glossina) transmit deadly trypanosomes to human populations and domestic animals in sub-Saharan Africa. Some foci of Human African Trypanosomiasis due to Trypanosoma brucei gambiense (g-HAT) persist in southern Chad, where a program of tsetse control was implemented against the local vector Glossina fuscipes fuscipes in 2018 in Maro. We analyzed the population genetics of G. f. fuscipes from the Maro focus before control (T0), one year (T1), and 18 months (T2) after the beginning of control efforts. Most flies captured displayed a local genetic profile (local survivors), but a few flies displayed outlier genotypes. Moreover, disturbance of isolation by distance signature (increase of genetic distance with geographic distance) and effective population size estimates, absence of any genetic signature of a bottleneck, and an increase of genetic diversity between T0 and T2 strongly suggest gene flows from various origins, and a limited impact of the vector control efforts on this tsetse population. Continuous control and surveillance of g-HAT transmission is thus recommended in Maro. Particular attention will need to be paid to the border with the Central African Republic, a country where the entomological and epidemiological status of g-HAT is unknown.

Title: Impact limité de la lutte antivectorielle sur la structure des populations de Glossina fuscipes fuscipes dans le foyer de la maladie du sommeil de Maro, Tchad. Abstract: Les mouches tsé-tsé (genre Glossina) transmettent des trypanosomes mortels aux populations humaines ainsi qu'aux animaux domestiques en Afrique sub-saharienne. Certains foyers de la trypanosomiase humaine Africaine due à Trypanosoma brucei gambiense (THA-g) persistent au sud du Tchad, où un programme de lutte antivectorielle a été mis en place contre le vecteur local de la maladie, Glossina fuscipes fuscipes, en particulier à Maro en 2018. Nous avons analysé la structure génétique des populations de G. f. fuscipes de ce foyer à T0 (avant lutte), une année après le début de la lutte (T1), et 18 mois après (T2). La plupart des mouches capturées après le début de la lutte ont montré un profil génétique local (survivants locaux), mais quelques-unes d'entre elles présentaient des génotypes d'individus atypiques. Par ailleurs, la présence de perturbations des signatures d'isolement par la distance (augmentation de la distance génétique avec la distance géographique), l'absence de signature génétique d'un goulot d'étranglement, et un accroissement de la diversité génétique entre T0 et T2 sont des arguments forts en faveur de la recolonisation de la zone par des mouches d'origines variées, tout en témoignant des effets limités de la campagne de lutte dans ce foyer. Ces résultats conduisent à recommander une lutte et une surveillance continues dans le foyer de Maro. Une attention particulière devra par ailleurs être prêtée à l'autre côté de la rive, située côté République Centre Africaine, dont le statut épidémiologique reste inconnu concernant les tsé-tsé et la THA-g.

Conducting active screening for human African trypanosomiasis with rapid diagnostic tests: The Guinean experience (2016-2021).

Strategies to detect Human African Trypanosomiasis (HAT) cases rely on serological screening of populations exposed to trypanosomes. In Guinea, mass medical screening surveys performed with the Card Agglutination Test for Trypanosomiasis have been progressively replaced by door-to-door approaches using Rapid Diagnostic Tests (RDTs) since 2016. However, RDTs availability represents a major concern and medical teams must often adapt, even in the absence of prior RDT performance evaluation. For the last 5 years, the Guinean HAT National Control Program had to combine three different RDTs according to their availability and price: the SD Bioline HAT (not available anymore), the HAT Sero-K-SeT (most expensive), and recently the Abbott Bioline HAT 2.0 (limited field evaluation). Here, we assess the performance of these RDTs, alone or in different combinations, through the analysis of both prospective and retrospective data. A parallel assessment showed a higher positivity rate of Abbott Bioline HAT 2.0 (6.0%, n = 2,250) as compared to HAT Sero-K-SeT (1.9%), with a combined positive predictive value (PPV) of 20.0%. However, an evaluation of Abbott Bioline HAT 2.0 alone revealed a low PPV of 3.9% (n = 6,930) which was surpassed when using Abbott Bioline HAT 2.0 in first line and HAT Sero-K-SeT as a secondary test before confirmation, with a combined PPV reaching 44.4%. A retrospective evaluation of all 3 RDTs was then conducted on 189 plasma samples from the HAT-NCP biobank, confirming the higher sensitivity (94.0% [85.6-97.7%]) and lower specificity (83.6% [76.0-89.1%]) of Abbott Bioline HAT 2.0 as compared to SD Bioline HAT (Se 64.2% [52.2-74.6%]-Sp 98.4% [94.2-99.5%]) and HAT Sero-K-SeT (Se 88.1% [78.2-93.8%]-Sp 98.4% [94.2-99.5%]). A comparison of Abbott Bioline HAT 2.0 and malaria-RDT positivity rates on 479 subjects living in HAT-free malaria-endemic areas further revealed that a significantly higher proportion of subjects positive in Abbott Bioline HAT 2.0 were also positive in malaria-RDT, suggesting a possible cross-reaction of Abbott Bioline HAT 2.0 with malaria-related biological factors in about 10% of malaria cases. This would explain, at least in part, the limited specificity of Abbott Bioline HAT 2.0. Overall, Abbott Bioline HAT 2.0 seems suitable as first line RDT in combination with a second HAT RDT to prevent confirmatory lab overload and loss of suspects during referral for confirmation. A state-of-the-art prospective comparative study is further required for comparing all current and future HAT RDTs to propose an optimal combination of RDTs for door-to-door active screening.

Gamma-radiation of Glossina palpalis gambiensis revisited: effect on fertility and mating competitiveness.

African animal trypanosomoses are vector-borne diseases that cause enormous livestock losses in sub-Saharan Africa, with drastic socio-economic impacts. Vector control in the context of an area-wide integrated pest management program with a sterile insect technique component requires the production of high-quality sterile male tsetse flies. In our study, we evaluated the effect of irradiation on the fecundity of Glossina palpalis gambiensis to identify the optimal dose that will induce maximum sterility while maintaining biological performance as much as possible. In addition, male mating performance was evaluated in semi-field cages. The irradiation doses used were 90, 100, 110, 120, 130, 140, and 150 Gy, and untreated males were used as the control. The results showed that pupal production and emergence rates were higher in batches of females that had mated with fertile males than in those that had mated with irradiated males with any experimental dose. A dose of 120 Gy administered to male flies induced 97-99% sterility after mating with virgin females. For the semi-field cage experiments, males irradiated with 120 Gy showed good sexual competitiveness as compared to fertile males and those irradiated with 140 Gy, considering the level of filling of spermatheca and the number of pairs formed. The optimal radiation dose of 120 Gy found in this study is slightly different from the traditional dose of 110 Gy that has been used in several eradication programmes in the past. The potential reasons for this difference are discussed, and an argument is made for the inclusion of reliable dosimetry systems in these types of studies.

Title: Le rayonnement gamma pour Glossina palpalis gambiensis revisité : effet sur la fertilité et la compétitivité sexuelle. Abstract: Les trypanosomoses animales africaines sont des maladies à transmission vectorielle qui causent d'énormes pertes de bétail en Afrique subsaharienne, avec des impacts socio-économiques importants. La lutte antivectorielle dans le cadre d'un programme de lutte intégrée contre les ravageurs à l'échelle d'une zone avec une composante de technique d'insectes stériles nécessite la production de glossines mâles stériles de haute qualité. Dans notre étude, nous avons évalué l'effet de l'irradiation sur la fécondité de Glossina palpalis gambiensis afin d'identifier la dose optimale qui induira une stérilité maximale tout en maintenant au maximum les performances biologiques. De plus, les performances d'accouplement des mâles ont été évaluées en cages de semi-terrain. Les doses d'irradiation utilisées étaient de 90, 100, 110, 120, 130, 140 et 150 Gy, et des mâles non traités ont été utilisés comme contrôle. Les résultats ont montré que les taux de production et d'émergence de pupes étaient plus élevés dans les lots de femelles qui s'étaient accouplées avec des mâles fertiles que dans les lots de celles accouplées avec des mâles irradiés, avec n'importe quelle dose expérimentale. Une dose de 120 Gy administrée à des mouches mâles a induit une stérilité de 97 à 99 % après accouplement avec des femelles vierges. Pour les expériences en cages de semi-terrain, les mâles irradiés à 120 Gy ont montré une bonne compétitivité sexuelle par rapport aux mâles fertiles et à ceux irradiés à 140 Gy, en considérant le niveau de remplissage de leur spermathèque et le nombre de couples formés. La dose de rayonnement optimale de 120 Gy trouvée dans cette étude est légèrement différente de la dose traditionnelle de 110 Gy qui a été utilisée dans plusieurs programmes d'éradication dans le passé. Les raisons potentielles de cette différence sont discutées et un argument est avancé pour l'inclusion de systèmes de dosimétrie fiables dans ce type d'études.

Infection of the malaria vector Anopheles coluzzii with the entomopathogenic bacteria Chromobacterium anophelis sp. nov. IRSSSOUMB001 reduces larval survival and adult reproductive potential.

BACKGROUND: Vector control tools are urgently needed to control malaria transmission in Africa. A native strain of Chromobacterium sp. from Burkina Faso was recently isolated and preliminarily named Chromobacterium anophelis sp. nov. IRSSSOUMB001. In bioassays, this bacterium showed a promising virulence against adult mosquitoes and reduces their blood feeding propensity and fecundity. The current study assessed the entomopathogenic effects of C. anophelis IRSSSOUMB001 on larval stages of mosquitoes, as well as its impacts on infected mosquitoes reproductive capacity and trans-generational effects. METHODS: Virulence on larvae and interference with insemination were assayed by co-incubation with C. anophelis IRSSSOUMB001 at a range of 104 to 108 cfu/ml. Trans-generational effects were determined by measuring body size differences of progeny from infected vs. uninfected parent mosquitoes using wing size as a proxy. RESULTS: Chromobacterium anophelis IRSSSOUMB001 killed larvae of the pyrethroid-resistant Anopheles coluzzii with LT80 of ~ 1.75 ± 0.14 days at 108 cfu/ml in larval breeding trays. Reproductive success was reduced as a measure of insemination rate from 95 ± 1.99% to 21 ± 3.76% for the infected females. There was a difference in wing sizes between control and infected mosquito offsprings from 2.55 ± 0.17 mm to 2.1 ± 0.21 mm in infected females, and from 2.43 ± 0.13 mm to 1.99 ± 0.15 mm in infected males. CONCLUSIONS: This study showed that C. anophelis IRSSSOUMB001 was highly virulent against larvae of insecticide-resistant Anopheles coluzzii, and reduced both mosquito reproduction capacity and offspring fitness. Additional laboratory, field, safety and social acceptance studies are needed to draw firm conclusions about the practical utility of this bacterial strain for malaria vector control.

Proof-of-concept study for a long-acting formulation of ivermectin injected in cattle as a complementary malaria vector control tool.

BACKGROUND: Domesticated animals play a role in maintaining residual transmission of Plasmodium parasites of humans, by offering alternative blood meal sources for malaria vectors to survive on. However, the blood of animals treated with veterinary formulations of the anti-helminthic drug ivermectin can have an insecticidal effect on adult malaria vector mosquitoes. This study therefore assessed the effects of treating cattle with long-acting injectable formulations of ivermectin on the survival of an important malaria vector species, to determine whether it has potential as a complementary vector control measure. METHODS: Eight head of a local breed of cattle were randomly assigned to either one of two treatment arms (2 × 2 cattle injected with one of two long-acting formulations of ivermectin with the BEPO® technology at the therapeutic dose of 1.2 mg/kg), or one of two control arms (2 × 2 cattle injected with the vehicles of the formulations). The lethality of the formulations was evaluated on 3-5-day-old Anopheles coluzzii mosquitoes through direct skin-feeding assays, from 1 to 210 days after treatment. The efficacy of each formulation was evaluated and compared using Cox proportional hazards survival models, Kaplan-Meier survival estimates, and log-logistic regression on cumulative mortality. RESULTS: Both formulations released mosquitocidal concentrations of ivermectin until 210 days post-treatment (hazard ratio > 1). The treatments significantly reduced mosquito survival, with average median survival time of 4-5 days post-feeding. The lethal concentrations to kill 50% of the Anopheles (LC50) before they became infectious (10 days after an infectious blood meal) were maintained for 210 days post-injection for both formulations. CONCLUSIONS: This long-lasting formulation of ivermectin injected in cattle could complement insecticide-treated nets by suppressing field populations of zoophagic mosquitoes that are responsible, at least in part, for residual malaria transmission. The impact of this approach will of course depend on the field epidemiological context. Complementary studies will be necessary to characterize ivermectin withdrawal times and potential environmental toxicity.

Analysis of the Genetic Variation of the Fruitless Gene within the Anopheles gambiae (Diptera: Culicidae) Complex Populations in Africa.

Targeting genes involved in sexual determinism, for vector or pest control purposes, requires a better understanding of their polymorphism in natural populations in order to ensure a rapid spread of the construct. By using genomic data from An. gambiae s.l., we analyzed the genetic variation and the conservation score of the fru gene in 18 natural populations across Africa. A total of 34,339 SNPs were identified, including 3.11% non-synonymous segregating sites. Overall, the nucleotide diversity was low, and the Tajima's D neutrality test was negative, indicating an excess of low frequency SNPs in the fru gene. The allelic frequencies of the non-synonymous SNPs were low (freq < 0.26), except for two SNPs identified at high frequencies (freq > 0.8) in the zinc-finger A and B protein domains. The conservation score was variable throughout the fru gene, with maximum values in the exonic regions compared to the intronic regions. These results showed a low genetic variation overall in the exonic regions, especially the male sex-specific exon and the BTB-exon 1 of the fru gene. These findings will facilitate the development of an effective gene drive construct targeting the fru gene that can rapidly spread without encountering resistance in wild populations.

Magnitude and associated factors of latent tuberculosis infection due to Mycobacterium tuberculosis complex among high-risk groups in urban Bobo-Dioulasso, Burkina Faso.

Objectives: To determine the prevalence and risk factors for latent tuberculosis infection (LTBI) among three high-risk groups - household contacts of TB index cases, healthcare workers and slaughterhouse workers - in Bobo-Dioulasso, Burkina Faso. Methods: Participants were recruited to this cross-sectional study from March to July 2020 after giving informed consent. Sociodemographic, clinical and biological data were collected using a structured questionnaire. The QuantiFERON-TB Gold Plus test (QFT-Plus) and the tuberculin skin test (TST) were used for detection of LTBI. Bivariate and multivariate logistic regression analyses were performed to identify risk factors for LTBI. Results: The prevalence of LTBI among 101 participants (age range 15-68 years) was 67.33% [95% confidence interval (CI) 57.27-76.33] and 84.16% (95% CI 75.55-90.66) based on QFT-Plus and TST results, respectively. Compared with healthcare workers and household contacts of TB index cases, the prevalence of LTBI among slaughterhouse workers was significantly higher for both QTF-Plus (96.8%; P<0.001) and TST (100%; P=0.003). Working in a slaughterhouse [adjusted odds ratio (AOR) 1.095, 95% CI 1.00-2.036], smoking (AOR 4.214, 95% CI 1.051-16.899), ≥15 years of exposure (AOR 5.617, 95% CI 1.202-32.198), having an animal at home (AOR 2.735, 95% CI 1.102-6.789) and protozoal infection (AOR 2.591, 95% CI 1.034-6.491) were significantly associated with LTBI on the QFT-Plus assay. Conclusion: The prevalence of LTBI was high in all three groups, particularly slaughterhouse workers. The risk factors identified could form the basis of targeted intervention.

Evaluation of antibody responses to tsetse fly saliva in domestic animals in the sleeping sickness endemic foci of Bonon and Sinfra, Côte d'Ivoire.

After intensive control efforts, human African trypanosomiasis (HAT) was declared eliminated in Côte d'Ivoire as a public health problem in December 2020 and the current objective is to achieve the interruption of the transmission (zero cases). Reaching this objective could be hindered by the existence of an animal reservoir of Trypanosoma (T.) brucei (b.) gambiense. In the framework of a study led in 2013 to assess the role of domestic animals in the epidemiology of HAT in the two last active foci from Côte d'Ivoire (Bonon and Sinfra), plasmas were sampled from four species of domestic animals for parasitological (microscopic examination by the buffy coat technique (BCT)), serological (immune trypanolysis (TL)) and molecular (specific PCR: TBR for T. brucei s.l., TCF for T. congolense forest type, TVW for T. vivax and PCR for T. b. gambiense) testing. In order to improve the understanding of the involvement/role of these animals in the transmission of T. b. gambiense, we have quantified in this study the IgG response to whole saliva extracts of Glossina palpalis gambiensis in order to perform an association analysis between anti-saliva responses and the positivity of diagnostic tests. Cattle and pigs had significantly higher rates of anti-tsetse saliva responses compared to goats and sheep (p < 0.01). In addition, the anti-tsetse saliva responses were strongly associated with the parasitology (BCT+), serology (TL+) and PCR (TBR+ and TCF+) results (p < 0.001). These associations indicate a high level of contacts between the positive/infected animals and tsetse flies. Our findings suggest that protecting cattle and pigs against tsetse bites could have a significant impact in reducing transmission of both animal and human trypanosome species, and advocates for a "One health" approach to better control African trypanosomosis in Côte d'Ivoire.

Monitoring the elimination of gambiense human African trypanosomiasis in the historical focus of Batié, South-West Burkina Faso.

The World Health Organisation has targeted the elimination of human African trypanosomiasis (HAT) as zero transmission by 2030. Continued surveillance needs to be in place for early detection of re-emergent cases. In this context, the performance of diagnostic tests and testing algorithms for detection of the re-emergence of Trypanosoma brucei gambiense HAT remains to be assessed. We carried out a door-to-door active medical survey for HAT in the historical focus of Batié, South-West Burkina Faso. Screening was done using three rapid diagnostic tests (RDTs). Two laboratory tests (ELISA/T. b. gambiense and immune trypanolysis) and parasitological examination were performed on RDT positives only. In total, 5883 participants were screened, among which 842 (14%) tested positive in at least one RDT. Blood from 519 RDT positives was examined microscopically but no trypanosomes were observed. The HAT Sero-K-Set test showed the lowest specificity of 89%, while the specificities of SD Bioline HAT and rHAT Sero-Strip were 92% and 99%, respectively. The specificity of ELISA/T. b. gambiense and trypanolysis was 99% (98-99%) and 100% (99-100%), respectively. Our results suggest that T. b. gambiense is no longer circulating in the study area and that zero transmission has probably been attained. While a least cost analysis is still required, our study showed that RDT preselection followed by trypanolysis may be a useful strategy for post-elimination surveillance in Burkina Faso.

Title: Suivi de l'élimination de la Trypanosomiase Humaine Africaine dans le foyer historique de Batié au sud-ouest du Burkina Faso. Abstract: L'Organisation mondiale de la santé a ciblé l'élimination de la trypanosomiase humaine africaine (THA) comme transmission zéro d'ici 2030. Une surveillance continue doit être mise en place pour la détection précoce des cas réémergents. Dans ce contexte, la performance des tests de diagnostic et des algorithmes de test pour la détection de la réémergence de la THA de Trypanosoma brucei gambiense reste à évaluer. Nous avons réalisé une enquête médicale en porte-à-porte pour la THA dans le foyer historique de Batié, au sud-ouest du Burkina Faso. Le dépistage a été effectué à l'aide de trois tests de diagnostic rapide (TDR). Deux tests de laboratoire (ELISA/T. b. gambiense et trypanolyse immunitaire) et un examen parasitologique ont été effectués uniquement sur les TDR positifs. Au total, 5883 participants ont été dépistés, parmi lesquels 842 (14 %) ont été testés positifs dans au moins un TDR. Le sang de 519 TDR positifs a été examiné au microscope mais aucun trypanosome n'a été observé. Le test HAT Sero-K-Set a montré la spécificité la plus faible de 89 %, tandis que les spécificités de SD Bioline HAT et rHAT Sero-Strip étaient de 92 % et 99 %, respectivement. La spécificité d'ELISA/T. b. gambiense et de la trypanolyse étaient respectivement de 99 % (98­99 %) et 100 % (99­100 %). Nos résultats suggèrent que T. b. gambiense ne circule plus dans la zone d'étude et que la transmission zéro a probablement été atteinte. Bien qu'une analyse de moindre coût soit toujours nécessaire, notre étude a montré qu'une présélection par TDR suivie d'une trypanolyse peut être une stratégie utile pour la surveillance post-élimination au Burkina Faso.

Evaluation of the Re-emergence Risk of Human African Trypanosomiasis in the Southwestern Burkina Faso, A Gold-Bearing Mutation Area.

PURPOSE: The boom in Burkina Faso's artisanal gold mining since 2007 has attracted populations from Côte d'Ivoire and Guinea, which are the West African countries most affected by human African trypanosomiasis (HAT) and therefore increases its risk of re-emergence. Our aim was to update the HAT data in Burkina Faso in the risk of the re-emergence context with the advent of artisanal gold mining. METHODS: The study was carried out in the southwestern Burkina Faso where entomological surveys were conducted using biconical traps in March 2017. Follow by an active medical survey in April 2017, which was targeted the gold panners in 7 villages closer to artisanal gold sites, using CATT, mini-anion exchange centrifugation technique, trypanolysis test (TL) and ELISA test to measure human/tsetse contacts. The buffy coat technique and the TL were also applied in pigs to check their reservoir role of human trypanosomes. RESULTS: Our results have shown no case of HAT among 958 individuals tested and all the 50 pigs were also negative, but the level of antibodies against tsetse saliva evidenced by ELISA revealed low human/tsetse contact. Moreover, gold panners practise agriculture and breeding in an infected tsetse area, which are increased the risk. CONCLUSION: Our results illustrate that the risk of re-emergence is low. The passive surveillance system implemented in 2015 in southwestern Burkina Faso is needed to increase the sentinel sites to better cover this area by taking into account the gold mining. Finally, awareness-raising activities are needed among populations about HAT.

Novel protein candidates for serodiagnosis of African animal trypanosomosis: Evaluation of the diagnostic potential of lysophospholipase and glycerol kinase from Trypanosoma brucei.

African trypanosomosis, a parasitic disease caused by protozoan parasites transmitted by tsetse flies, affects both humans and animals in sub-Saharan Africa. While the human form (HAT) is now limited to foci, the animal form (AAT) is widespread and affects the majority of sub-Saharan African countries, and constitutes a real obstacle to the development of animal breeding. The control of AAT is hampered by a lack of standardized and easy-to used diagnosis tools. This study aimed to evaluate the diagnostic potential of TbLysoPLA and TbGK proteins from Trypanosoma brucei brucei for AAT serodiagnosis in indirect ELISA using experimental and field sera, individually, in combination, and associated with the BiP C-terminal domain (C25) from T. congolense. These novel proteins were characterized in silico, and their sequence analysis showed strong identities with their orthologs in other trypanosomes (more than 60% for TbLysoPLA and more than 82% for TbGK). TbLysoPLA displays a low homology with cattle (<35%) and Piroplasma (<15%). However, TbGK shares more than 58% with cattle and between 45-55% with Piroplasma. We could identify seven predicted epitopes on TbLysoPLA sequence and 14 potential epitopes on TbGK. Both proteins were recombinantly expressed in Escherichia coli. Their diagnostic potential was evaluated by ELISA with sera from cattle experimentally infected with T. congolense and with T.b. brucei, sera from cattle naturally infected with T. congolense, T. vivax and T.b. brucei. Both proteins used separately had poor diagnostic performance. However, used together with the BiP protein, they showed 60% of sensitivity and between 87-96% of specificity, comparable to reference ELISA tests. In conclusion, we showed that the performance of the protein combinations is much better than the proteins tested individually for the diagnosis of AAT.

Cross border transhumance involvement in ticks and tick-borne pathogens dissemination and first evidence of Anaplasma centrale in Burkina Faso.

In West Africa, cross-border transhumance, also called seasonal migration, is known to be a very important animal production strategy, as it involves about 70 to 90% of cattle. In spite of the cattle movements, some strategic areas of transhumance remain poorly explored regarding ticks and their associated pathogens investigations. The purpose of this study is to evaluate the involvement of transhumance in the spread of cattle ticks and associated pathogens in Burkina Faso (BF) and Benin (BN), in a context of speedy invasion of West African livestock by Rhipicephalus microplus. A longitudinal survey was performed on 210 cattle from BF, monitored for ticks and tick-borne pathogens (TBP) during one seasonal transhumance. The first sampling coded "T0BF" took place in eastern BF, at the transhumance departure. A second sampling "T1BN" was carried out in northern BN, the transhumance arrival zone. A third sampling "T2BF" was done at the return of cattle in eastern BF. Ticks were morphologically identified and TBP detected with reverse line blot hybridization (RLB) assay. A total of 1027 ticks (7 species), 1006 ticks (11 species) and 1211 ticks (9 species) were respectively found at T0BF, T1BN and T2BF. Some species were collected at the three times of sampling without any significant difference in their relative abundances. However, other tick species appeared only at T1BN and/or T2BF. The TBP species found at the three points surveyed were Theileria annulata, Theileria mutans, Theileria velifera, Babesia bigemina and Anaplasma marginale. The most prevalent was T. mutans with 166/210 (79%), 159/210 (75.7%) and 78/210 (37%) cattle positive respectively at T0BF, T1BN and T2BF. Anaplasma centrale was evidenced with 0.5% and 0.9% respectively at T0BF and T2BF. To our knowledge, this represents its first report in the study area. Overall, the TBP prevalences were significantly lower at T2BF, highlighting the effect of tick populations changes induced by transhumance combined with the seasonal variation influence.

Assessment of novel Lehmann's funnel entry trap prototypes performance to control malaria mosquito populations.

BACKGROUND: There is a global consensus that new intervention tools are needed for the final steps toward malaria elimination/eradication. In a recent study in Burkina Faso, the Lehmann Funnel Entry Trap (LFET) has shown promising results in the reduction of mosquito densities, even in areas where insecticide resistance is as high as 80%. The LFET requires no chemicals and is self-operated. However, one of the issues with the original LFET is the size of the funnel, which often occupies too much space within users' homes. Here, the performance of three new, smaller-sized LFET prototypes that combine a screening and killing effect on mosquitoes was assessed. METHODS: The study was carried out over three months during the rainy season in low and high malaria vector density sites, Soumousso and Vallée du Kou, respectively. The original LFET (or 'Prototype 1'/'P1') was modified to produce three new prototypes, which were referred to as prototype 2 ('the Medium' or 'P2'), prototype 3 (P3) and prototype 4 (P4). Each of the new prototypes was tested on eight days per month over the three-month period to assess their effectiveness in trapping and killing mosquitoes entering houses through the windows compared to the original LFET. RESULTS: Overall, 78,435 mosquitoes (mainly Anopheles gambiae sensu lato) were collected in the two study sites, both in the traps and in the houses. A total of 56,430 (72%) mosquitoes were collected from the traps. In Vallée du Kou, the original LFET caught a greater number of mosquitoes than the medium (prototype 2), whereas no difference was observed between the other new prototypes (3 and 4) and the medium. In Soumousso, both the original and medium LFETs collected significantly greater numbers of mosquitoes compared to prototypes 3 and 4. CONCLUSION: This study has shown that the new LFET prototypes are effective in trapping mosquitoes in high mosquito density settings. A large-scale study with one of the prototypes will be needed to assess community acceptance of the traps and their ability to control malaria vectors.

Optimizing the feeding frequency to maximize the production of sterile males in tsetse mass-rearing colonies.

Tsetse flies are cyclical vectors of trypanosomes, the causative agents of sleeping sickness or Human African Trypanosomosis and nagana or African Animal Trypanosomosis in Sub-Saharan Africa. The Insectarium de Bobo-Dioulasso (IBD) was created and equipped in the frame of Pan African Tsetse and Trypanosomosis Eradication Campaign (PATTEC) with the main goal to provide sterile males for the different eradication programs in West Africa which is already the case with the ongoing eradication program in Senegal. The aim of this study was to identify the best feeding regime in mass-rearing colonies of Glossina palpalis gambiensis to optimize the yield of sterile males. We investigated the mortality and fecundity for various feeding regimes and day alternation (3×: Monday-Wednesday-Friday, 4×: Monday-Wednesday-Friday-Saturday, 4×: Monday-Wednesday-Thursday-Friday and 6×: all days except Sunday) on adult tsetse flies in routine rearing over 60 days after emergence. The day alternation in the 4 blood meals per week (feeding regimes 2 and 3) had no effect on tsetse fly mortality and fecundity. The best feeding regime was the regime of 4 blood meals per week which resulted in higher significant fecundity (PPIF = 2.5; P = 0.003) combined with lower mortality of females (P = 0.0003) than the 3 blood meals per week (PPIF = 2.0) and in similar fecundity (PPIF = 2.6; P = 0.70) and mortality (P = 0.51) than the 6 blood meals per week. This feeding regime was extended to the whole colonies, resulting in an improved yield of sterile males for the ongoing eradication program in Senegal and would be more cost-effective for the implementation of the next-coming sterile insect technique (SIT) programs in West Africa.

Macrophage migrating inhibitory factor expression is associated with Trypanosoma brucei gambiense infection and is controlled by trans-acting expression quantitative trait loci in the Guinean population.

Infection by Trypanosoma brucei gambiense is characterized by a wide array of clinical outcomes, ranging from asymptomatic to acute disease and even spontaneous cure. In this study, we investigated the association between macrophage migrating inhibitory factor (MIF), an important pro-inflammatory cytokine that plays a central role in both innate and acquired immunity, and disease outcome during T. b. gambiense infection. A comparative expression analysis of patients, individuals with latent infection and controls found that MIF had significantly higher expression in patients (n = 141; 1.25 ±â€¯0.07; p < .0001) and latent infections (n = 25; 1.23 ±â€¯0.13; p = .0005) relative to controls (n = 46; 0.94 ±â€¯0.11). Furthermore, expression decreased significantly after treatment (patients before treatment n = 33; 1.40 ±â€¯0.18 versus patients after treatment n = 33; 0.99 ±â€¯0.10, p = .0001). We conducted a genome wide eQTL analysis on 29 controls, 128 cases and 15 latently infected individuals for whom expression and genotype data were both available. Four loci, including one containing the chemokine CXCL13, were found to associate with MIF expression. Genes at these loci are candidate regulators of increased expression of MIF after infection. Our study is the first data demonstrating that MIF expression is elevated in T. b. gambiense-infected human hosts but does not appear to contribute to pathology.

Drug quality analysis of isometamidium chloride hydrochloride and diminazene diaceturate used for the treatment of African animal trypanosomosis in West Africa.

BACKGROUND: Diminazene diaceturate (DA) and isometamidium chloride hydrochloride (ISM) are with homidium bromide, the main molecules used to treat African Animal Trypanosomosis (AAT). These drugs can be purchased from official suppliers but also from unofficial sources like local food markets or street vendors. The sub-standard quality of some of these trypanocides is jeopardizing the efficacy of treatment of sick livestock, leading thus to economic losses for the low-resource farmers and is contributing to the emergence and spread of drug resistance. The objective of this study was to assess the quality of trypanocidal drugs sold in French speaking countries of West Africa. In total, 308 drug samples including 282 of DA and 26 of ISM were purchased from official and unofficial sources in Benin, Burkina Faso, Côte d'Ivoire, Mali, Niger and Togo. All samples were analysed at LACOMEV (Dakar, Senegal), a reference laboratory of the World Organisation for Animal Health, by galenic inspection and high performance liquid chromatography. RESULTS: The results showed that 51.90% of the samples were non-compliant compared to the standards and were containing lower quantity of the active ingredient compared to the indications on the packaging. The non-compliances ranged from 63.27% in Togo to 32.65% in Burkina Faso (61.82% in Benin, 53.84% in Mali, 50% in Côte d'Ivoire, 47.36% in Niger). The rates of non-compliance were not statistically different (P = 0.572) from official or unofficial suppliers and ranged from 30 to 75% and from 0 to 65% respectively. However, the non-compliance was significantly higher for ISM compared to DA (P = 0.028). CONCLUSIONS: The high non-compliance revealed in this study compromises the efficacy of therapeutic strategies against AAT, and is likely to exacerbate chemoresistance in West Africa. Corrective actions against sub-standard trypanocides urgently need to be taken by policy makers and control authorities.

Differences in pathogenicity and virulence of Trypanosoma brucei gambiense field isolates in experimentally infected Balb/C mice.

Trypanosoma brucei gambiense (T. b. gambiense) is the major causative agent of human African trypanosomiasis (HAT). A great variety of clinical outcomes have been observed in West African foci, probably due to complex host-parasite interactions. In order to separate the roles of parasite genetic diversity and host variability, we have chosen to precisely characterize the pathogenicity and virulence of T. b. gambiense field isolates in a mouse model. Thirteen T. b. gambiense strains were studied in experimental infections, with 20 Balb/C infected mice per isolate. Mice were monitored for 30 days, in which mortality, parasitemia, anemia, and weight were recorded. Mortality rate, prepatent period, and maximum parasitemia were estimated, and a survival analysis was performed to compare strain pathogenicity. Mixed models were used to assess parasitemia dynamics, weight, and changes in Packed Cell Volume (PCV). Finally, a multivariate analysis was performed to infer relationships between all variables. A large phenotypic diversity was observed. Pathogenicity was highly variable, ranging from strains that kill their host within 9 days to a non-pathogenic strain (no deaths during the experiment). Virulence was also variable, with maximum parasitemia values ranging from 42 million to 1 billion trypanosomes/ml. Reduced PCV and weight occurred in the first two weeks of the infection, with the exception of two strains. Finally, the global analysis highlighted three groups of strains: a first group with highly pathogenic strains showing an early mortality associated with a short prepatent period; a second group of highly virulent strains with intermediate pathogenicity; and a third group of isolates characterized by low pathogenicity and virulence patterns. Such biological differences could be related to the observed clinical diversity in HAT. A better understanding of the biological pathways underlying the observed phenotypic diversity could thus help to clarify the complex nature of the host-parasite interactions that determine the resistance/susceptibility status to T. brucei gambiense.

Candidate gene polymorphisms study between human African trypanosomiasis clinical phenotypes in Guinea.

BACKGROUND: Human African trypanosomiasis (HAT), a lethal disease induced by Trypanosoma brucei gambiense, has a range of clinical outcomes in its human host in West Africa: an acute form progressing rapidly to second stage, spontaneous self-cure and individuals able to regulate parasitaemia at very low levels, have all been reported from endemic foci. In order to test if this clinical diversity is influenced by host genetic determinants, the association between candidate gene polymorphisms and HAT outcome was investigated in populations from HAT active foci in Guinea. METHODOLOGY AND RESULTS: Samples were collected from 425 individuals; comprising of 232 HAT cases, 79 subjects with long lasting positive and specific serology but negative parasitology and 114 endemic controls. Genotypes of 28 SNPs in eight genes passed quality control and were used for an association analysis. IL6 rs1818879 allele A (p = 0.0001, OR = 0.39, CI95 = [0.24-0.63], BONF = 0.0034) was associated with a lower risk of progressing from latent infection to active disease. MIF rs36086171 allele G seemed to be associated with an increased risk (p = 0.0239, OR = 1.65, CI95 = [1.07-2.53], BONF = 0.6697) but did not remain significant after Bonferroni correction. Similarly MIF rs12483859 C allele seems be associated with latent infections (p = 0.0077, OR = 1.86, CI95 = [1.18-2.95], BONF = 0.2157). We confirmed earlier observations that APOL1 G2 allele (DEL) (p = 0.0011, OR = 2.70, CI95 = [1.49-4.91], BONF = 0.0301) is associated with a higher risk and APOL1 G1 polymorphism (p = 0.0005, OR = 0.45, CI95 = [0.29-0.70], BONF = 0.0129) with a lower risk of developing HAT. No associations were found with other candidate genes. CONCLUSION: Our data show that host genes are involved in modulating Trypanosoma brucei gambiense infection outcome in infected individuals from Guinea with IL6 rs1818879 being associated with a lower risk of progressing to active HAT. These results enhance our understanding of host-parasite interactions and, ultimately, may lead to the development of new control tools.

[Risk assessment in young people living in Bobo Dioulasso: analysis of factors associated with sexual precocity and multiple partners]. / Prises de risques chez les jeunes de Bobo Dioulasso: une analyse des facteurs associés à la précocité et au multipartenariat sexuel.

INTRODUCTION: Despite health education efforts, young people continue to adopt risky sexual behaviors which may have a significant impact on their health. This study aims to analyze the factors associated with sexual precocity and multiple partners among young people aged 19-24 years living in Bobo-Dioulasso. METHODS: We conducted a quantitative, cross-sectional study. Survey data were collected from 573 young people aged 15-24 years in Bobo-Dioulasso (Burkina Faso) in December 2014. The respondents were selected using two stages cluster sampling. Risk factors for sexual precocity and multiple partners were analyzed using Stata 13 IC software. We used P<0.05 as the significance level. RESULTS: More than half of respondents (54%) were sexually active, of whom 14% before the age of 16. Having multiple sex partners was reported by 24% of sexually active youth. Determinants of sexual precocity and multiple partners were age, sex, level of education and economic status of parents. Our data have also shown that early sexual debut was associated with multiple sexual partners (p<0.005). CONCLUSION: Actions aimed to convince young people to delay sexual debut and to better assess risks are of utmost importance. Enhancing parents, teachers and carers capabilities is essential to improve the quality of their relationships with young people.

Population Genetics and Reproductive Strategies of African Trypanosomes: Revisiting Available Published Data.

Trypanosomatidae are a dangerous family of Euglenobionta parasites that threaten the health and economy of millions of people around the world. More precisely describing the population biology and reproductive mode of such pests is not only a matter of pure science, but can also be useful for understanding parasite adaptation, as well as how parasitism, specialization (parasite specificity), and complex life cycles evolve over time. Studying this parasite's reproductive strategies and population structure can also contribute key information to the understanding of the epidemiology of associated diseases; it can also provide clues for elaborating control programs and predicting the probability of success for control campaigns (such as vaccines and drug therapies), along with emergence or re-emergence risks. Population genetics tools, if appropriately used, can provide precise and useful information in these investigations. In this paper, we revisit recent data collected during population genetics surveys of different Trypanosoma species in sub-Saharan Africa. Reproductive modes and population structure depend not only on the taxon but also on the geographical location and data quality (absence or presence of DNA amplification failures). We conclude on issues regarding future directions of research, in particular vis-à-vis genotyping and sampling strategies, which are still relevant yet, too often, neglected issues.