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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.

In Subsaharan Africa, tsetse flies (genus Glossina) are vectors of trypanosomes causing Human African Trypanosomiasis (HAT) and Animal African Trypanosomosis (AAT). Some foci of HAT persist in Southern Chad, where a program of tsetse control was started against the local vector Glossina fuscipes fuscipes in the Mandoul focus in 2014, and in Maro in 2018. Flies were also sampled in 2018 in Timbéri and Dokoutou. We analyzed the population genetics of G. fuscipes fuscipes from the four tsetse-infested zones. The trapping samples were characterized by a strong female biased sex-ratio, except in Timbéri and Dokoutou that had high tsetse densities. Apparent density and effective population density appeared smaller in the main foci of Mandoul and Maro and the average dispersal distance (within the spatial scale of each zone) was as large as or larger than the total length of each respective zone. The genetic signature of a population bottleneck was found in the Mandoul and Timbéri area, suggesting a large ancient interconnected metapopulation that underwent genetic subdivision into small, isolated pockets due to adverse environmental conditions. The long-range dispersal and the existence of genetic outliers suggest a possibility of migration from remote sites such as the Central African Republic in the south (although the fly situation remains unknown there) and/or a genetic signature of recent exchanges. Due to likely isolation, an eradication strategy may be considered for sustainable HAT control in Mandoul focus. Another strategy will probably be required in Maro focus, which probably experiences much more exchanges with its neighbors.

Background: Human African trypanosomiasis (HAT) is a neglected tropical disease caused by Trypanosoma brucei gambiense transmitted by tsetse flies in sub-Saharan West Africa. In southern Chad the most active and persistent focus is the Mandoul focus, with 98% of the reported human cases, and where African animal trypanosomosis (AAT) is also present. Recently, a control project to eliminate tsetse flies (Glossina fuscipes fuscipes) in this focus using the sterile insect technique (SIT) was initiated. However, the release of large numbers of sterile males of G. f. fuscipes might result in a potential temporary increase in transmission of trypanosomes since male tsetse flies are also able to transmit the parasite. The objective of this work was therefore to experimentally assess the vector competence of sterile males treated with isometamidium for Trypanosoma brucei brucei. Methods: An experimental infection was set up in the laboratory, mimicking field conditions: the same tsetse species that is present in Mandoul was used. A T. b. brucei strain close to T. b. gambiense was used, and the ability of the sterile male tsetse flies fed on blood with and without a trypanocide to acquire and transmit trypanosomes was measured. Results: Only 2% of the experimentally infected flies developed an immature infection (midgut) while none of the flies developed a metacyclic infection of T. b. brucei in the salivary glands. We did not observe any effect of the trypanocide used (isometamidium chloride at 100 mg/l) on the development of infection in the flies. Conclusions: Our results indicate that sterile males of the tested strain of G. f. fuscipes were unable to cyclically transmit T. b. brucei and might even be refractory to the infection. The data of the research indicate that the risk of cyclical transmission of T. brucei by sterile male G. f. fuscipes of the strain colonized at IAEA for almost 40 years appears to be small.

La maladie du sommeil, ou trypanosomiase humaine africaine (THA), est due à Trypanosoma brucei transmis par la glossine ou mouche tsé-tsé. Après avoir ravagé l'Afrique subsaharienne dans la première moitié du XXe siècle, comme en témoigne l'extraordinaire ouvrage d'Alphonse Laveran et Félix Mesnil en 1912 (Trypanosomes et trypanosomiases), la THA, maladie tropicale négligée mortelle, dite " du bout de la piste " et pour laquelle il n'existe pas de vaccin, semble aujourd'hui à portée de main de l'élimination. Nous passons en revue les stratégies, activités et outils qui ont permis cette forte réduction du fardeau sanitaire pour les populations d'Afrique subsaharienne : environ 300 000 cas estimés dans les années 1990 contre moins de 1000 cas rapportés annuellement depuis 2018. La lutte contre la maladie du sommeil consiste principalement à dépister et traiter les cas ainsi qu'à lutter contre la glossine vectrice pour casser le cycle de transmission. En passant en 30 ans d'un contexte épidémique à un contexte d'élimination, la maladie du sommeil a subi une transition épidémiologique sans précédent à laquelle les stratégies et les outils de lutte ont dû s'adapter. Nous montrons comment la recherche a soutenu ces efforts et examinons certains des défis restants pour rendre effective et durable son élimination.

Livestock are seasonally subjected to the nuisance of haematophagous flies, such as tabanids and stomoxyine flies. Topical application of insecticides has short term efficacy (a week or so), is expensive, and generates pesticide residues in animal products and environment. Attractive insecticide-impregnated blue fabrics are used for tsetse fly control in Africa; however, they are expensive and were never evaluated for other haematophagous flies. In previous works, we defined specifications of a white and blue screen specifically attracting haematophagous flies, particularly Stomoxys spp. In the present study, an assay was carried out in Kantchanaburi Province, Thailand, with around 30 of such screen prototypes, made of a multilayer polyethylene film incorporated with deltamethrin. Screens (also called 'targets') were deployed in 12 test farms, to evaluate the efficacy of a so-called 'multi-target method' (MTM); four control farms were also enrolled. A Vavoua trap was deployed one day/week in each farm to follow-up the density of insects. In the test-farms, during the 4 months post treatment, the mean density of haematophagous flies was significantly and consistently reduced by 63-73% compared to the control group. Laboratory tests indicated that insecticidal activity of these screen prototypes lasted around 3-4 months. However, in the field, significant reduction of fly densities was observed in all test farms up to 7 months after screen deployment, possibly as a consequence of the early impact of the screens on fly population dynamics. The significant effects obtained in test farms provided evidence for the proof of concept that MTM is effective for on-farm control of haematophagous and common flies. Durability of the screens will be increased in the next prototype generation. This innovative control method will be evaluated more extensively and in other livestock and poultry farms.

Background: The sterile insect technique (SIT) requires mass-rearing of the target species, irradiation to induce sexual sterility and transportation from the mass-rearing facility to the target site. Those treatments require several steps that may affect the biological quality of sterile males. This study has been carried out to evaluate the relative impact of chilling, irradiation and transport on emergence rate, flight ability and survival of sterile male Glossina palpalis gambiensis. Results: Chilling, irradiation and transport all affected the quality control parameters studied. The emergence rate was significantly reduced by long chilling periods and transport, i.e. from 92% at the source insectary in Burkina Faso to 78% upon arrival in Senegal. Flight ability was affected by all three parameters with 31% operational flies lost between the production facility and the destination site. Only survival under stress was not affected by any of the treatments. Conclusion: The chilling period and transport were the main factors that impacted significantly the quality of sterile male pupae. Therefore, in the operational programme, the delivery of sterile male pupae was divided over two shipments per week to reduce the chilling time and improve the quality of the sterile males. Quality of the male pupae may further be improved by reducing the transport time and vibrations during transport.

Background: Tsetse flies are vectors of African trypanosomes, protozoan parasites that cause sleeping sickness (or human African trypanosomosis) in humans and nagana (or animal African trypanosomosis) in livestock. In addition to trypanosomes, four symbiotic bacteria Wigglesworthia glossinidia, Sodalis glossinidius, Wolbachia, Spiroplasma and one pathogen, the salivary gland hypertrophy virus (SGHV), have been reported in different tsetse species. We evaluated the prevalence and coinfection dynamics between Wolbachia, trypanosomes, and SGHV in four tsetse species (Glossina palpalis gambiensis, G. tachinoides, G. morsitans submorsitans, and G. medicorum) that were collected between 2008 and 2015 from 46 geographical locations in West Africa, i.e. Burkina Faso, Mali, Ghana, Guinea, and Senegal. Results: The results indicated an overall low prevalence of SGHV and Wolbachia and a high prevalence of trypanosomes in the sampled wild tsetse populations. The prevalence of all three infections varied among tsetse species and sample origin. The highest trypanosome prevalence was found in Glossina tachinoides (61.1%) from Ghana and in Glossina palpalis gambiensis (43.7%) from Senegal. The trypanosome prevalence in the four species from Burkina Faso was lower, i.e. 39.6% in Glossina medicorum, 18.08%; in Glossina morsitans submorsitans, 16.8%; in Glossina tachinoides and 10.5% in Glossina palpalis gambiensis. The trypanosome prevalence in Glossina palpalis gambiensis was lowest in Mali (6.9%) and Guinea (2.2%). The prevalence of SGHV and Wolbachia was very low irrespective of location or tsetse species with an average of 1.7% for SGHV and 1.0% for Wolbachia. In some cases, mixed infections with different trypanosome species were detected. The highest prevalence of coinfection was Trypanosoma vivax and other Trypanosoma species (9.5%) followed by coinfection of T. congolense with other trypanosomes (7.5%). The prevalence of coinfection of T. vivax and T. congolense was (1.0%) an

In order to assess the effect of irradiation on male pupae of Glossina palpalis gambiensis, we assessed quality parameters of the irradiated male pupae, notably the hatching rate of the pupae, the flying ability of emerged tsetse flies and survival rate of young flies in starving condition following an available quality control protocol. The fertility of male flies from irradiated pupae was also assessed through female productivity and the insemination ability of irradiated males. For this purpose 30 virgin females of 3 days old were mated with 10 irradiated males aged 6 days in breeding cages. Productivity (number of pupae) was compared to a control batch. Simultaneously to this test, other cages of 30 virgin females of 3 days old, mated with 10 irradiated males of 6 days old were followed for the insemination capacity of the irradiated males. Dissections were performed to evaluate the filling level of spermatheca of females mated with irradi-ated males and those mated with control males. Irradiation had no effect on pupae hatching (p=0.083). The fly ability rate was significantly greater in the control group than in the treated lot (p<0.001). Male survival from control pupae was significantly longer (p<0.001), with an average of 5 and 4 days respectively for the control and ir-radiated treatment. Female productivity was 1.67 ± 1.55 pupae and 18.93 ± 3.38 pupae, respectively, for females mated with irradiated males and females mated with control males, respectivily. The sterility rate induced in females was 89.67%. Most females had full spermatheca and spermatheca fill level was generally similar for both treatments. Ultimately, irradiation does not affect the viability of the pupae. However, it has an ef-fect on the performance of male flies without affecting their biological quality.

In many parts of Africa, tsetse eradication is impossible due to political, environmental or economic circumstances. In these situations, African animal trypanosomosis control relies on communities or farmer-based control, implemented at a local scale in accordance to the eco- epidemiological context and the cattle rearing system to be sustainable. Management of the African animal trypanosomosis requires integrated controls strategies that combine the use of more than one locally-based tool and where possible, needs to be assisted by veterinarians and other animal health professionals. Several tsetse control methods based on insecticide treated cattle (i.e. pour-on, manual spraying, community bath) and insecticide treated target (traps and screens impregnated with insecticides) are available and should be complemented with diagnostic tests and medication (active trypanocides with prophylactic and/or therapeutic action). However, their adoption is mainly dependent on the engagement of communities, farmers and herders. Indeed, the adoption of a locally-adapted control strategy will depend on farmers socio-technical networks, the cost-effectiveness of the control activities, as well as the time and cost for implementation. In general, insecticide treated cattle methods are the most suitable and acceptable for farmers, because they protect a private good i.e. cattle, whereas insecticide treated targets are generally considered to provide a public good. Nonetheless, selection of the most appropriate tools requires consideration of local disease epidemiology (including host-parasite coevolution), local environmental and socio-economic constraints. The active involvement of communities, farmers and herders is essential from the beginning of the conception of innovative control strategies, and the cost of local integrated pest management should be reduced as much as possible, to be adopted as an acceptable and sustainable animal production cost.

The Government of Senegal has embarked several years ago on a project that aims to eradicate Glossina palpalis gambiensis from the Niayes area. The removal of the animal trypanosomosis would allow the development more efficient livestock production systems. The project was implemented using an area-wide integrated pest management strategy including a sterile insect technique (SIT) component. The released sterile male flies originated from a colony from Burkina Faso. Methodology/Principal Findings Monitoring the efficacy of the sterile male releases requires the discrimination between wild and sterile male G. p. gambiensis that are sampled in monitoring traps. Before being released, sterile male flies were marked with a fluorescent dye powder. The marking was however not infallible with some sterile flies only slightly marked or some wild flies contaminated with a few dye particles in the monitoring traps. Trapped flies can also be damaged due to predation by ants, making it difficult to discriminate between wild and sterile males using a fluorescence camera and / or a fluorescence microscope. We developed a molecular technique based on the determination of cytochrome oxidase haplotypes of G. p. gambiensis to discriminate between wild and sterile males. DNA was isolated from the head of flies and a portion of the 5' end of the mitochondrial gene cytochrome oxidase I was amplified to be finally sequenced. Our results indicated that all the sterile males from the Burkina Faso colony displayed the same haplotype and systematically differed from wild male flies trapped in Senegal and Burkina Faso. This allowed 100% discrimination between sterile and wild male G. p. gambiensis. Conclusions/Significance This tool might be useful for other tsetse control campaigns with a SIT component in the framework of the Pan-African Tsetse and Trypanosomosis Eradication Campaign (PATTEC) and, more generally, for other vector or insect pest control programs.

Background: Tsetse flies transmit trypanosomes that cause human and African animal trypanosomosis, a debilitating disease of humans (sleeping sickness) and livestock (nagana). An area-wide integrated pest management campaign against Glossina palpalis gambiensis has been implemented in Senegal since 2010 that includes a sterile insect technique (SIT) component. The SIT can only be successful when the sterile males that are destined for release have a flight ability, survival and competitiveness that are as close as possible to that of their wild male counterparts. Methodology/Principal Findings: Tests were developed to assess the quality of G. p. gambiensis males that emerged from pupae that were produced and irradiated in Burkina Faso and Slovakia (irradiation done in Seibersdorf, Austria) and transported weekly under chilled conditions to Dakar, Senegal. For each consignment a sample of 50 pupae was used for a quality control test (QC group). To assess flight ability, the pupae were put in a cylinder filtering emerged flies that were able to escape the cylinder. The survival of these flyers was thereafter monitored under stress conditions (without feeding). Remaining pupae were emerged and released in the target area of the eradication programme (RF group). The following parameter values were obtained for the QC flies: average emergence rate more than 69%, median survival of 6 days, and average flight ability of more than 35%. The quality protocol was a good proxy of fly quality, explaining a large part of the variances of the examined parameters. Conclusions/Significance: The quality protocol described here will allow the accurate monitoring of the quality of shipped sterile male tsetse used in operational eradication programmes in the framework of the Pan-African Tsetse and Trypanosomosis Eradication Campaign.

Background: The application of the sterile insect technique (SIT) requires mass-production of sterile males of good biological quality. The size of the project area will in most cases determine whether it is more cost effective to produce the sterile flies locally (and invest in a mass-rearing facility) or import the sterile flies from a mass-rearing facility that is located in another country. This study aimed at assessing the effect of long distance transport of sterile male Glossina palpalis gambiensis pupae on adult male fly yield. Methods: The male pupae were produced at the Centre International de Recherche-Développement sur l'Elevage en zone Subhumide (CIRDES), Bobo-Dioulasso, Burkina Faso, and shipped with a commercial courier service in insulated transport boxes at a temperature of ±10°C to Senegal (±36 h of transport). Upon arrival in the insectary in Dakar, the pupae were transferred to an emergence room and the flies monitored for 3-6 days. Results: The results showed that the used system of isothermal boxes that contained phase change material packs (S8) managed to keep the temperature at around 10°C which prevented male fly emergence during transport. The emergence rate was significantly higher for pupae from batch 2 (chilled at 4°C for one day in the source insectary before transport) than those from batch 1 (chilled at 4°C for two days in the source insectary before transport) i.e. an average (±sd) of 76.1 ± 13.2% and 72.2 ± 14.3%, respectively with a small proportion emerging during transport (0.7 ± 1.7% and 0.9 ± 2.9%, respectively). Among the emerged flies, the percentage with deformed (not fully expanded) wings was significantly higher for flies from batch 1 (12.0 ± 6.3%) than from batch 2 (10.7 ± 7.5%). The amount of sterile males available for release as a proportion of the total pupae shipped was 65.8 ± 13.3% and 61.7 ± 14.7% for batch 1 and 2 pupae, respectively. Conclusions: The results also showed that the temperature inside.