Glossina Palpalis Research Articles

Thioester-containing proteins in the tsetse fly (Glossina) and their response to trypanosome infection.

Thioester‐containing proteins (TEPs) are conserved proteins with a role in innate immune immunity. In the current study, we characterized the TEP family in the genome of six tsetse fly species (Glossina spp.). Tsetse flies are the biological vectors of several African trypanosomes, which cause sleeping sickness in humans or nagana in livestock. The analysis of the tsetse TEP sequences revealed information about their structure, evolutionary relationships and expression profiles under both normal and trypanosome infection conditions. Phylogenetic analysis of the family showed that tsetse flies harbour a genomic expansion of specific TEPs that are not found in other dipterans. We found a general expression of all TEP genes in the alimentary tract, mouthparts and salivary glands. Glossina morsitans and Glossina palpalis TEP genes display a tissue‐specific expression pattern with some that are markedly up‐regulated when the fly is infected with the trypanosome parasite. A different TEP response was observed to infection with Trypanosoma brucei compared to Trypanosoma congolense, indicating that the tsetse TEP response is trypanosome‐specific. These findings are suggestive for the involvement of the TEP family in tsetse innate immunity, with a possible role in the control of the trypanosome parasite.

Prevalence of Sodalis glossinidius and different trypanosome species in Glossina palpalis palpalis caught in the Fontem sleeping sickness focus of the southern Cameroon.

Tsetse flies are the cyclical vector of human and animal African trypanosomiasis. To improve vector control in order to achieve the elimination of human African trypanosomiasis (HAT) and boost the control of animal diseases, investigations have been undertaken on the tripartite association between tsetse, trypanosome, and symbionts. It is in this light that Sodalis glossinidius and different trypanosomes were identified in Glossina palpalis palpalis caught in Fontem in southern Cameroon. For this study, DNA was extracted from whole flies, and S. glossinidius and different trypanosome species were identified by polymerase chain reaction (PCR). Statistical analyses were performed to compare the trypanosome and S. glossinidius infection rates and to look for an association between these microorganisms. Of the 274 G. p. palpalis caught, 3.3% (9/274) were teneral. About 35% (96/274) of these flies harbored S. glossinidius. Of the 265 non-teneral flies, 37.7% were infected by trypanosomes. The infection rates of Trypanosoma congolense “forest type” and Trypanosoma vivax were 26.04% and 18.11%, respectively. About 6.41% of tsetse harbored mixed infections of T. congolense and T. vivax. Of the 69 tsetse with T. congolense infections, 33.33% (23/69) harbored S. glossinidius while 71.86% (69/96) of flies harboring S. glossinidius were not infected by trypanosomes. No association was observed between S. glossinidius and trypanosome infections. Some wild tsetse harbor S. glossinidius and trypanosomes, while others have no infection or are infected by only one of these microorganisms. We conclude that the presence of S. glossinidius does not favor trypanosome infections in G. p. palpalis of the Fontem focus.

Competitiveness and survival of two strains of Glossina palpalis gambiensis in an urban area of Senegal

BackgroundIn the Niayes area, located in the west of Senegal, only one tsetse species, Glossina palpalis gambiensis Vanderplank (Diptera: Glossinidae) was present. The Government of Senegal initiated and implemented an elimination programme in this area that included a sterile insect technique (SIT) component. The G. p. gambiensis strain (BKF) mass-reared at the Centre International de Recherche-Développement sur l'Elevage en zone Subhumide (CIRDES) in Burkina Faso was used for the SIT component.Methodology/principal findingsStudies conducted in 2011 in four localities in the Niayes area (Pout, Sébikotane, Diacksao Peul and the Parc de Hann) showed that the BKF strain demonstrated inferior survival in the ecosystem of the Parc de Hann, a forested area in the city centre of the capital Dakar. Therefore, G. p. gambiensis flies from the Niayes area (SEN strain) were colonized. Here we compared the competitiveness and survival of the two strains (BKF and SEN) in the Parc de Hann. Released sterile males of the SEN colony showed a daily mortality rate of 0.08 (SD 0.08) as compared with 0.14 (SD 0.08) for the BKF flies but the difference was not significant (p-value = 0.14). However, the competitiveness of the SEN males was lower (0.14 (SD 0.10)) as compared with that of the BKF males (0.76 (SD 0.11)) (p-value < 10−3).Conclusions/significanceBased on the results of this study, it can be concluded that the BKF strain will remain the main strain to be used in the elimination programme. Despite the slightly longer survival of the SEN males in the Parc de Hann, the superior competitiveness of the BKF males is deemed more important for the SIT component, as their shorter survival rates can be easily compensated for by more frequent fly releases.

Fipronil 1% pour-on: further studies of its effects against lab-reared Glossina palpalis gambiensis

In order to assess the residual effects of fipronil 1% on tsetse fly survival, male Glossina palpalis gambiensis were released on non-treated and treated cattle, with 0.1 ml of fipronil/kg b.w. as a pour-on formulation. In a second trial, the female fecundity performances were evaluated by feeding teneral females on the same cattle. These females were then mated and their production parameters monitored, as well as the survival of freshly emerged flies. Fipronil had a significant effect on tsetse fly survival (p < 0.001). Over a period of 30 days, up to 40% of tsetse fly mortality was observed within 72 h after tsetse were released. The residual effects ranged between 51 and 74 days when tsetse flies were released twice within a 15-day interval in the presence of a treated animal. When tsetse flies were fed on treated cattle through a parafilm membrane, 92 days after the treatment, no significant effect of fipronil was observed on the reproductive performance of females, i.e., as well as on fecundity (p = 0.948) and emergence rates (p = 0.743), or puparial weight (p = 0.422). This was also the case for the survival of young flies, with no difference observed between the two groups. After this study, it is confirmed that fipronil is highly effective against tsetse flies. Its efficacy in controlling ticks is already known but other externalities such as the control of biting insects add value to its use.

Intestinal Bacterial Communities of Trypanosome-Infected and Uninfected Glossina palpalis palpalis from Three Human African Trypanomiasis Foci in Cameroon.

Glossina sp. the tsetse fly that transmits trypanosomes causing the Human or the Animal African Trypanosomiasis (HAT or AAT) can harbor symbiotic bacteria that are known to play a crucial role in the fly's vector competence. We hypothesized that other bacteria could be present, and that some of them could also influence the fly's vector competence. In this context the objectives of our work were: (a) to characterize the bacteria that compose the G. palpalis palpalis midgut bacteriome, (b) to evidence possible bacterial community differences between trypanosome-infected and non-infected fly individuals from a given AAT and HAT focus or from different foci using barcoded Illumina sequencing of the hypervariable V3-V4 region of the 16S rRNA gene. Forty G. p. palpalis flies, either infected by Trypanosoma congolense or uninfected were sampled from three trypanosomiasis foci in Cameroon. A total of 143 OTUs were detected in the midgut samples. Most taxa were identified at the genus level, nearly 50% at the species level; they belonged to 83 genera principally within the phyla Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria. Prominent representatives included Wigglesworthia (the fly's obligate symbiont), Serratia, and Enterobacter hormaechei. Wolbachia was identified for the first time in G. p. palpalis. The average number of bacterial species per tsetse sample was not significantly different regarding the fly infection status, and the hierarchical analysis based on the differences in bacterial community structure did not provide a clear clustering between infected and non-infected flies. Finally, the most important result was the evidence of the overall very large diversity of intestinal bacteria which, except for Wigglesworthia, were unevenly distributed over the sampled flies regardless of their geographic origin and their trypanosome infection status.

Transcriptional Profiling of Midguts Prepared from Trypanosoma/T. congolense-Positive Glossina palpalis palpalis Collected from Two Distinct Cameroonian Foci: Coordinated Signatures of the Midguts' Remodeling As T. congolense-Supportive Niches.

Our previous transcriptomic analysis of Glossina palpalis gambiensis experimentally infected or not with Trypanosoma brucei gambiense aimed to detect differentially expressed genes (DEGs) associated with infection. Specifically, we selected candidate genes governing tsetse fly vector competence that could be used in the context of an anti-vector strategy, to control human and/or animal trypanosomiasis. The present study aimed to verify whether gene expression in field tsetse flies (G. p. palpalis) is modified in response to natural infection by trypanosomes (T. congolense), as reported when insectary-raised flies (G. p. gambiensis) are experimentally infected with T. b. gambiense. This was achieved using the RNA-seq approach, which identified 524 DEGs in infected vs. non-infected tsetse flies, including 285 downregulated genes and 239 upregulated genes (identified using DESeq2). Several of these genes were highly differentially expressed, with log2 fold change values in the vicinity of either +40 or −40. Downregulated genes were primarily involved in transcription/translation processes, whereas encoded upregulated genes governed amino acid and nucleotide biosynthesis pathways. The BioCyc metabolic pathways associated with infection also revealed that downregulated genes were mainly involved in fly immunity processes. Importantly, our study demonstrates that data on the molecular cross-talk between the host and the parasite (as well as the always present fly microbiome) recorded from an experimental biological model has a counterpart in field flies, which in turn validates the use of experimental host/parasite couples.

Comparative Genomics of Glossina palpalis gambiensis and G. morsitans morsitans to Reveal Gene Orthologs Involved in Infection by Trypanosoma brucei gambiense.

Blood-feeding Glossina palpalis gambiense (Gpg) fly transmits the single-celled eukaryotic parasite Trypanosoma brucei gambiense (Tbg), the second Glossina fly African trypanosome pair being Glossina morsitans/T.brucei rhodesiense. Whatever the T. brucei subspecies, whereas the onset of their developmental program in the zoo-anthropophilic blood feeding flies does unfold in the fly midgut, its completion is taking place in the fly salivary gland where does emerge a low size metacyclic trypomastigote population displaying features that account for its establishment in mammals-human individuals included. Considering that the two Glossina—T. brucei pairs introduced above share similarity with respect to the developmental program of this African parasite, we were curious to map on the Glossina morsitans morsitans (Gmm), the Differentially Expressed Genes (DEGs) we listed in a previous study. Briefly, using the gut samples collected at days 3, 10, and 20 from Gpg that were fed or not at day 0 on Tbg—hosting mice, these DGE lists were obtained from RNA seq—based approaches. Here, post the mapping on the quality controlled DEGs on the Gmm genome, the identified ortholog genes were further annotated, the resulting datasets being compared. Around 50% of the Gpg DEGs were shown to have orthologs in the Gmm genome. Under one of the three Glossina midgut sampling conditions, the number of DEGs was even higher when mapping on the Gmm genome than initially recorded. Many Gmm genes annotated as “Hypothetical” were mapped and annotated on many distinct databases allowing some of them to be properly identified. We identify Glossina fly candidate genes encoding (a) a broad panel of proteases as well as (b) chitin—binding proteins, (c) antimicrobial peptide production—Pro3 protein, transferrin, mucin, atttacin, cecropin, etc—to further select in functional studies, the objectives being to probe and validated fly genome manipulation that prevents the onset of the developmental program of one or the other T. brucei spp. stumpy form sampled by one of the other bloodfeeding Glossina subspecies.

Using genetic data to improve species distribution models

Tsetse flies (Diptera, Glossinidae) transmit human and animal trypanosomoses in Africa, respectively a neglected human disease (sleeping sickness) and the most important constraint to cattle production in infested countries (nagana). We recently developed a methodology to map landscape friction (i.e. resistance to movement) for tsetse in West Africa. The goal was to identify natural barriers to tsetse dispersal, and potentially isolated tsetse populations for targeting elimination programmes. Most species distribution models neglect landscape functional connectivity whereas environmental factors affecting suitability or abundance are not necessarily the same as those influencing gene flows. Geographic distributions of a given species can be seen as the intersection between biotic (B), abiotic (A) and movement (M) factors (BAM diagram).Here we show that the suitable habitat for Glossina palpalis gambiensis as modelled by Maxent can be corrected by landscape functional connectivity (M) extracted from our friction analysis. This procedure did not degrade the specificity of the distribution model (P=0.751) whereas the predicted distribution area was reduced. The added value of this approach is that it reveals unconnected habitat patches.The approach we developed on tsetse to inform landscape connectivity (M) is reproducible and does not rely on expert knowledge. It can be applied to any species: we call for a generalization of the use of M to improve distribution models.

The Dynamics of Tsetse Flies Around the Mbam and Djerem National Park

A capture of tsetse flies was done between November 2014 and July 2016 in three localities around the Mbam and Djerem National Park. The biconic traps were used for the captures in Yoko, Mbakaou and Betare-Oya during 5 days. These traps permitted to capture 709 tsetse flies belonging to four species: Glossina fuscipes fuscipes (54.16%), Glossina fusca congolensis (29.34%), Glossina palpalis spp (7.19%) and Glossina calliginea (9.31%). The season did not influence the abundance of tsetse flies. In general, no significant influence was observed between the localities. Specifically, the Glossina palpalis spp species presented a significant difference in its distribution between the localities of Betare-Oya and Yoko (P=0.048). The Glossina calliginea species was also differently distributed between the traps close to a path in T4 enlightened area and the traps close to a path to T3 shade (P=0.042), presenting a direct link between the number of tsetse flies and the presence of cattle.

Influence of temperature and relative humidity on survival and fecundity of three tsetse strains.

BackgroundTsetse flies occur in much of sub-Saharan Africa where they are vectors of trypanosomes that cause human and animal African trypanosomosis. The sterile insect technique (SIT) is currently used to eliminate tsetse fly populations in an area-wide integrated pest management (AW-IPM) context in Senegal and Ethiopia. Three Glossina palpalis gambiensis strains [originating from Burkina Faso (BKF), Senegal (SEN) and an introgressed strain (SENbkf)] were established and are now available for use in future AW-IPM programmes against trypanosomes in West Africa. For each strain, knowledge of the environmental survival thresholds is essential to determine which of these strains is best suited to a particular environment or ecosystem, and can therefore be used effectively in SIT programmes.MethodsIn this paper, we investigated the survival and fecundity of three G. p. gambiensis strains maintained under various conditions: 25 °C and 40, 50, 60, and 75 % relative humidity (rH), 30 °C and 60 % rH and 35 °C and 60 % rH.ResultsThe survival of the three strains was dependent on temperature only, and it was unaffected by changing humidity within the tested range. The BKF strain survived temperatures above its optimum better than the SEN strain. The SENbkf showed intermediate resistance to high temperatures. A temperature of about 32 °C was the limit for survival for all strains. A rH ranging from 40 to 76 % had no effect on fecundity at 25–26 °C.ConclusionsWe discuss the implications of these results on tsetse SIT-based control programmes.

Eco-epidemiology of porcine trypanosomosis in Karim Lamido, Nigeria: prevalence, seasonal distribution, tsetse density and infection rates.

BackgroundAnimal trypanosomosis is a major economic disease in Nigeria causing considerable morbidity and mortality in livestock. Despite reports in other animals, the disease is under reported in pigs.MethodsWe conducted a community based epidemiological study on African animal trypanosomosis in Karim Lamido area of Taraba State, Nigeria using species-specific PCR on 712 pigs and 706 of the 2822 captured tsetse flies. Data were analysed using Chi-square, odds ratio and multivariate analysis at 95 % confidence interval.ResultsOverall prevalence of porcine trypanosomosis was 16.6 % and ranged between 2.0 and 8.8 % across Trypanosoma species. Seasonal distribution of porcine trypanosomosis varied significantly (χ2 = 16.62, df = 3, P = 0.0008) ranging between 7.9 and 23.6 % across seasons. Mixed infections involving T. b. brucei, T. congolense forest and T. congolense savannah recorded infection rates ranging between 2.5 and 9.3 %. There were significant variations between the trypanosome infection rates in relation to age (χ2 = 7.629, df = 1, P = 0.0057, OR = 1.932, 95 % CI = 1.203–3.100), sex (χ2 = 10.09, df = 1, P = 0.0015, OR = 2.085, 95 % CI = 1.315–3.304) and body condition (χ2 = 22.10, df = 2, P < 0.0001) of pigs ranging between 10.4 and 30.3 %. Tsetse infection rates were 11.2 % (79/706) for Glossina palpalis and 6.8 % (48/706) for G. tachinoides yielding an overall infection rate of 18.0 %.ConclusionTrypanosoma species are prevalent in the study area with similar distribution patterns in both pigs and tsetse flies. Late rainy season, adults, females and pigs with poor body condition recorded higher trypanosome infection rates. Of the three Trypanosoma spp. identified, T. b. brucei showed predominance.

Variations in attack behaviours between Glossina palpalis gambiensis and G. tachinoides in a gallery forest suggest host specificity.

Tsetse flies Glossina palpalis gambiensis and G. tachinoides are among the major vectors of sleeping sickness (Human African Trypanosomiasis-HAT) and nagana (African Animal Trypanosomiasis-AAT) in West Africa. Both riparian species occur sympatrically in gallery forests of south west Burkina Faso, but little is known of their interspecies relationships although different authors think there may be some competition between them. The aim of this study was to check if sympatric species have different strategies when approaching a host. A man placed in a sticky cube (1 m × 1 m × 1 m) and a sticky black-blue-black target (1 m × 1 m) were used to capture tsetse along the Comoe river banks in a Latin Square design. The number and the height at which tsetse were caught by each capture method were recorded according to species and sex. Glossina p. gambiensis was more attracted to human bait than to the target, but both species were captured at a significantly higher height on the target compared with the human bait (P < 0.05). No significant difference in heights was found between G. tachinoides and G. p. gambiensis captured on targets (33 and 35 cm, respectively, P > 0.05). However, catches on human bait showed a significant difference in height between G. tachinoides and G. p. gambiensis (22.5 and 30.6 cm, respectively, P < 0.001). This study showed that these sympatric species had different attack behaviours to humans, which is not the case with the target. The implications of these findings are discussed.

Molecular identification of different trypanosome species and subspecies in tsetse flies of northern Nigeria.

BackgroundAnimal African Trypanosomiasis (AAT) is caused by several species of trypanosomes including Trypanosoma congolense, T. vivax, T. godfreyi, T. simiae and T. brucei. Two of the subspecies of T. brucei also cause Human African Trypanosomiasis. Although some of them can be mechanically transmitted by biting flies; these trypanosomes are all transmitted by tsetse flies which are the cyclical vectors of Trypanosoma congolense, T. godfreyi, T. simiae and T. brucei. We present here the first report assessing the prevalence of trypanosomes in tsetse flies in Nigeria using molecular tools.Methods488 tsetse flies of three species, Glossina palpalis palpalis, G. tachinoides and G. morsitans submorsitans were collected from Wuya, Niger State and Yankari National Park, Bauchi State in 2012. Trypanosomes were detected and identified using an ITS1 PCR assay on DNA purified from the ‘head plus proboscis’ (H + P) and abdomen (ABD) parts of each fly.ResultsT. vivax and T. congolense Savannah were the major parasites detected. Trypanosomes prevalence was 7.1 % in G. p. palpalis, 11.9 % in G. tachinoides and 13.5 % in G. m. submorsitans. Prevalences of T. congolense Savannah ranged from 2.5 to 6.7 % and of T. vivax were approximately 4.5 %. Trypanosoma congolense Forest, T. godfreyi and T. simiae were also detected in the site of Yankari. The main biological and ecological determinants of trypanosome prevalence were the fly sex, with more trypanosomes found in females than males, and the site, with T. congolense subspp. being more abundant in Yankari than in Wuya. As expected, the trypanosome species diversity was higher in Yankari National Park than in the more agricultural site of Wuya where vertebrate host species diversity is lower.ConclusionsOur results show that T. congolense Savannah and T. vivax are the main species of parasite potentially causing AAT in the two study sites and that Yankari National Park is a potential reservoir of trypanosomes both in terms of parasite abundance and species diversity.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1585-3) contains supplementary material, which is available to authorized users.

Enquêtes entomologiques préparatoires à une lutte à grande échelle contre les glossines, assistées par un système d’information géographique : cas de la Pattec au Burkina Faso

Le système d’information géographique constitue une aide considérable lors de la définition d’un plan de lutte contre les glossines. Son application pratique pour la collecte de données entomologiques servant de base à la planification de la lutte et à son évaluation future a été faite dans l’ouest du Burkina Faso, de décembre 2007 à novembre 2008. Choisie pour englober les bassins versants des principaux fleuves ou rivières et les limites de distribution de Glossina palpalis gambiensis et G. tachinoides, la zone d’étude a été divisée en cellules de 10 km x 10 km. Dans chaque cellule, un maximum de 13 sites potentiels de piégeage ont été sélectionnés par l’utilisation d’imagerie satellite, puis repérés sur le terrain à l’aide du système mondial de positionnement (GPS). L’enquête entomologique a été réalisée avec des pièges biconiques Challier-Laveissière et les récoltes ont eu lieu 72 heures après leur installation. L’analyse des données a été conduite en utilisant des modèles aléa­toires mixtes binomiaux et de distribution de Poisson. La cartographie, réalisée avec le logiciel Arc Gis 9.3, a permis de déterminer la limite nord (12° 45’ N) des glos­sines dans la zone d’intervention. Les proportions de sites infestés étaient de 89,6 et 76,4 % respectivement au niveau des sections ascendante et descendante de la rivière Mouhoun (la première coule vers le nord-est depuis la source, la seconde vers le sud-est puis le sud après avoir conflué avec le Sourou), et de 16,7 et 10,3 % respectivement au niveau de leurs affluents. Entre les isohyètes 700–800 m et 800–900 m aucune différence significative n’a été observée dans les densités apparentes des glossines par piège, ni dans les sites infestés. Les deux espèces de glossines cap­turées étaient distribuées différemment selon les sections du Mouhoun : les cap­tures étaient composées de 79,5 % de G. p. gambiensis sur la section ascendante et de 96,0 % de G. tachinoides sur la section descendante. Les captures de vecteurs mécaniques ont été relativement faibles, leur distribution étant comparable à celle des glossines avec cependant quelques densités importantes sur les affluents.

Neemazal ® as a possible alternative control tool for malaria and African trypanosomiasis?

BackgroundResearch efforts to identify possible alternative control tools for malaria and African trypanosomiasis are needed. One promising approach relies on the use of traditional plant remedies with insecticidal activities.MethodsIn this study, we assessed the effect of blood treated with different doses of NeemAzal ® (NA, neem seed extract) on mosquitoes (Anopheles coluzzii) and tsetse flies (Glossina palpalis gambiensis) (i) avidity to feed on the treated blood, (ii) longevity, and (iii) behavioural responses to human and calf odours in dual-choice tests. We also gauged NeemAzal ® toxicity in mice.ResultsIn An. coluzzii, the ingestion of NA in bloodmeals offered by membrane feeding resulted in (i) primary antifeedancy; (ii) decreased longevity; and (iii) reduced response to host odours. In G. palpalis gambiensis, NA caused (i) a knock-down effect; (ii) decreased or increased longevity depending on the dose; and (iii) reduced response to host stimuli. In both cases, NA did not affect the anthropophilic rate of activated insects. Overall, the most significant effects were observed with NA treated bloodmeals at a dose of 2000 μg/ml for mosquitoes and 50 μg/ml for tsetse flies. Although no mortality in mice was observed after 14 days of follow-up at oral doses of 3.8, 5.6, 8.4 and 12.7 g/kg, behavioural alterations were noticed at doses above 8 g/kg.ConclusionThis study revealed promising activity of NA on A. coluzzii and G. palpalis gambiensis but additional research is needed to assess field efficacy of neem products to be possibly integrated in vector control programmes.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1538-x) contains supplementary material, which is available to authorized users.

Tsetse Control and the Elimination of Gambian Sleeping Sickness.

Tsetse Control and the Elimination of Gambian Sleeping Sickness.

Repartition Des Glossines Dans La Province De L’ogooue Ivindo Ancien Foyer De Trypanosomose Humaine Africaine

In Gabon, the situation of Human African Trypanosomosis (HAT) remains unclear. In addition, several historical foci existing in many provinces have not been explored for over 15 years. Yet many studies have shown that this country has to offer favorable ecosystems for the development of tsetse major vectors of HAT. To verify a risk of transmission of sleeping sickness, a tsetse inventory was conducted in the province of Ogooué Ivindo historic home of HAT. Vavoua and Nzi traps were placed in four characteristic habitats of the province: National Park Ivindo, Zadié Village, Forest of Zadié and baï (clearing) of Momba. A total of 2383 flies were captured: 1680 flies in the clearing of Momba, 437 flies in the primary forest Zadié, 139 in the National Park Ivindo and 12 in Zadié village. Moreover, these flies were divided into 7 species Glossina frezili, Glossina fusca congolensis, Glossina nashi, Glossina palpalis palpalis, Glossina tabaniformis, Glossina fuscipes fuscipes and Glossina tachinoides. Glossina palpalis palpalis (46%) was the most abundant species followed by Glossina fusca congolensis (21%) and Glossina nashi (16%). Glossina fuscipes fuscipes (6%), Glossina frezili (4%), Glossina tabaniformis (4%) and Glossina tachinoides (3%) were the least caught species. These results have shown that the province of Ogooué Ivindo still remains infested by tsetse flies. Also, further study monitoring these insects is essential to clarify the epidemiological importance of these vectors on human health in this province.

High Frequency of Trypanosoma congolense Savannah Type (Kinetoplastida: Trypanosomatidae) Among Tsetse Flies (Diptera: Glossinidae) in a Historic Trypanosoma Foci in North-Eastern Gabon: Preliminary Study [corrected

Human African trypanosomiasis became a neglected disease after the 1960s, when case numbers dropped dramatically. It again became a public health problem in sub-Saharan Africa at the end of the 1990s, when new cases were reported, notably in Central Africa, and specifically in Gabon, where historic foci existed and new cases have been reported. Therefore, the present study reports on an entomological survey conducted in May 2012 to determine the pathogenic trypanosome infection rate in tsetse flies and characterize the diversity of Trypanosoma species in the Ivindo National Park (INP) in northeastern Gabon. Nine Vavoua traps were used to catch tsetse over a 7-days period. All tsetse flies captured were identified to species, dissected, and trypanosome species identified using polymerase chain reaction (PCR). In total, 160 tsetse flies were analyzed, including Glossina palpalis palpalis, Glossina fusca congolense, and Glossina tachinoïdes The trypanosome infection rate of the flies was 6.3 and 31.9% using microscopy and PCR, respectively. The species identified were Trypanosoma congolense savannah type, Trypanosoma brucei brucei, Trypanosoma brucei gambiense, Trypanosoma vivax, and Trypanosoma congolense forest type. Trypanosoma risk index was 0.75 and 7.05 for humans and for animals, respectively. This study illustrates the diversity of Trypanosoma species infecting the tsetse flies in the INP. The simultaneous occurrence of Trypanosoma and tsetse from the palpalis group may suggest that the reservoirs of African animal trypanosomiasis should be carefully monitored in this area.

Xenomonitoring of sleeping sickness transmission in Campo (Cameroon).

BackgroundThe sleeping sickness focus of Campo in South Cameroon is still active, at a low endemic level, for more than a century, despite a regular medical surveillance. The present study focuses on the spatial distribution of xenomonitoring information obtained from an entomological survey performed in the dry season 2012. It appears that humans constitute a third of the blood meals and that the flies’ densities were coherent with those classically observed in the different biotopes. Paradoxically, the epicenter of the focus is the place where the risk indicators are the lowest ones.MethodsParticular attention was paid to the entomological device so that it covered the main part of human activities in the study area. One hundred and sixty-two pyramidal traps were used to catch tsetse flies twice a day that were identified, counted, dissected. Molecular analysis using classical and specific molecular markers was conducted to determine the importance of trypanosome infections and the nature of the feeding hosts. This information was used to calculate a Transmission Risk Index and to define a gradient of risk that was projected into a Geographical Information System.ResultsConventional entomological indicators such as species identification of tsetse flies or the Apparent Density per Trap per day, show that Glossina palpalis palpalis is the main species in the campo area which is classically distributed into the different biotopes of the study area. Molecular analysis reveals that humans constitute a third of the blood feeding hosts and that 20 % of the dissected flies were infected with trypanosomes, principally with Nannomonas. Nevertheless, one fly was carrying Trypanosoma brucei gambiense, the pathogen agent of sleeping sickness, showing that the reservoir is still active in the epicenter of the focus. Paradoxically, the Transmission Risk Index is not important in the epicenter, demonstrating that endemic events are not only depending on the man/vector contact.ConclusionXenomonitoring provides a valuable guide/tool to determine places at higher risk for vector/human contact and to identify trypanosomes species circulating in the focus. This information from xenomonitoring demonstrates that decision makers should include a veterinary device in a control strategy.

A Molecular Method to Discriminate between Mass-Reared Sterile and Wild Tsetse Flies during Eradication Programmes That Have a Sterile Insect Technique Component.

BackgroundThe 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 FindingsMonitoring 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/SignificanceThis 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.