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Background: African Trypanosomiases threaten the life of both humans and animals. Trypanosomes are transmitted by tsetse and other biting flies. In Rwanda, the African Animal Trypanosomiasis (AAT) endemic area is mainly around the tsetse-infested Akagera National Park (NP). The study aimed to identify Trypanosoma species circulating in cattle, their genetic diversity and distribution around the Akagera NP., Methodology: A cross-sectional study was carried out in four districts, where 1,037 cattle blood samples were collected. The presence of trypanosomes was determined by microscopy, immunological rapid test VerY Diag and PCR coupled with High-Resolution Melt (HRM) analysis. A parametric test (ANOVA) was used to compare the mean Packed cell Volume (PCV) and trypanosomes occurrence. The Cohen Kappa test was used to compare the level of agreement between the diagnostic methods., Findings: The overall prevalence of trypanosome infections was 5.6%, 7.1% and 18.7% by thin smear, Buffy coat technique and PCR/HRM respectively. Microscopy showed a low sensitivity while a low specificity was shown by the rapid test (VerY Diag). Trypanosoma (T.) congolense was found at a prevalence of 10.7%, T. vivax 5.2%, T. brucei brucei 2% and T. evansi 0.7% by PCR/HRM. This is the first report of T.evansi in cattle in Rwanda. The non-pathogenic T. theileri was also detected. Lower trypanosome infections were observed in Ankole x Friesian breeds than indigenous Ankole. No human-infective T. brucei rhodesiense was detected. There was no significant difference between the mean PCV of infected and non-infected animals (p>0.162)., Conclusions: Our study sheds light on the species of animal infective trypanosomes around the Akagera NP, including both pathogenic and non-pathogenic trypanosomes. The PCV estimation is not always an indication of trypanosome infection and the mechanical transmission should not be overlooked. The study confirms that the area around the Akagera NP is affected by AAT, and should, therefore, be targeted by the control activities. AAT impact assessment on cattle production and information on the use of trypanocides are needed to help policymakers prioritise target areas and optimize intervention strategies. Ultimately, these studies will allow Rwanda to advance in the Progressive Control Pathway (PCP) to reduce or eliminate the burden of AAT., Competing Interests: The authors have declared that no competing interests exist.

Background: African trypanosomiases are vector-borne diseases that affect humans and livestock in sub-Saharan Africa. Although data have been collected on tsetse fauna as well as trypanosome infections in tsetse flies and mammals in foci of sleeping sickness in Chad, the situation of tsetse fly-transmitted trypanosomes remains unknown in several tsetse-infested areas of Chad. This study was designed to fill this epidemiological knowledge gap by determining the tsetse fauna as well as the trypanosomes infecting tsetse flies in the area of Lake Iro in southeastern Chad., Methods: Tsetse flies were trapped along the Salamat River using biconical traps. The proboscis and tsetse body were removed from each fly. DNA was extracted from the proboscis using proteinase K and phosphate buffer and from the tsetse body using Chelex 5%. Tsetse flies were identified by amplifying and sequencing the cytochrome c oxydase I gene of each tsetse fly. Trypanosome species were detected by amplifying and sequencing the internal transcribed spacer 1 of infecting trypanosomes., Results: A total of 617 tsetse flies were trapped; the apparent density of flies per trap per day was 2. 6. Of the trapped flies, 359 were randomly selected for the molecular identification and for the detection of infecting trypanosomes. Glossina morsitans submorsitans (96.1%) was the dominant tsetse fly species followed by G. fuscipes fuscipes (3.1%) and G. tachinoides (0.8%). Four trypanosome species, including Trypanosoma vivax, T. simiae, T. godfreyi and T. congolense savannah, were detected. Both single infection (56.7%) and mixed infections of trypanosomes (4.6%) were detected in G. m. submorsitans. The single infection included T. simiae (20.5%), T. congolense savannah (16.43%), T. vivax (11.7%) and T. godfreyi (9.8%). The trypanosome infection rate was 61.4% in G. m. submorsitans, 72.7% in G. f. fuscipes and 66.6% in G. tachinoides. Trypanosome infections were more prevalent in tsetse bodies (40.6%) than in the proboscis (16.3%)., Conclusion: This study revealed the presence of different tsetse species and a diversity of trypanosomes pathogenic to livestock in the area of Lake Iro. The results highlight the risks and constraints that animal African trypanosomiasis pose to livestock breeding and the importance of assessing trypanosome infections in livestock in this area.

Tsetse-transmitted trypanosomosis remains a major animal health problem in Nigeria, in a context where changes in land cover, climate and control interventions are modifying its epidemiological patterns. Evidence-based decision making for the progressive control of the disease requires spatially-explicit information on its occurrence and prevalence, as well as on the distribution and abundance of the tsetse vector. In the framework of the continental Atlas of tsetse and African animal trypanosomosis (AAT), a geo-referenced database was assembled for Nigeria, based on the systematic review of 133 scientific publications (period January 1990 - March 2019). The three main species of trypanosomes responsible for the disease (i.e. Trypanosoma vivax, T. congolense and T. brucei) were found to be widespread, thus posing a national-level problem. Their geographic distribution extends beyond the tsetse-infested belt, owing to the combined effect of animal movement and mechanical transmission by non-tsetse vectors. T. simiae, the major trypanosomal pathogen in pigs, T. godfreyi and the human-infective T. brucei gambiense were also reported. AAT was reported in a number of susceptible host species, including cattle, sheep, goats, pigs, camels, horses, donkeys and dogs, while no study on wildlife was identified. Estimates of prevalence are heavily influenced by the sensitivity of the diagnostic techniques, ranging from an average of 3.5% for blood films to 31.0% for molecular techniques. Two riverine tsetse species (i.e. Glossina palpalis palpalis and G. tachinoides) were found to have the broadest geographical range, as they were detected in all six geopolitical zones of Nigeria. By contrast, the distribution of savannah species (i.e. G. morsitans submorsitans and G. longipalpis) appears to be highly fragmented, and limited to protected areas. Very little information is available for forest species, with one single paper reporting on G. fusca congolensis and G. nigrofusca nigrofusca in the Niger Delta region. The future development of a national Atlas of tsetse and AAT, relying on both published and unpublished information, could improve on the present review and provide further epidemiological evidence for decision making., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:, (Copyright © 2020 Food and Agriculture Organization of the United Nations (FAO). Published by Elsevier B.V. All rights reserved.)

Addressing the problems linked to tsetse-transmitted trypanosomiases requires considerable data on tsetse distribution and trypanosome infections. Although efforts to map tsetse and trypanosome infections have been undertaken at continental level, published data are still rare in wildlife reserves of West and Central Africa. To fill this gap, data on tsetse distribution and trypanosome infections were generated in the wildlife reserve of Santchou. For this study, each tsetse caught was identified and its DNA extracted. Different trypanosome species were identified by PCR. Entomological and parasitological data were transported onto a satellite image in order to visualize their distributions. From 195 Glossina palpalis palpalis that were caught, 33.8% (66/195) carried trypanosome infections with 89.4% (59/66) of single infections and 10.6% (7/66) mixed infections. From the 66 flies with trypanosome infections, 54.5% (36/66), 27.3% (18/66) and 18.2% (12/66) were respectively due to Trypanosoma congolense, Trypanosoma brucei s.l. and Trypanosoma vivax. The global infection rates were 18.5% (36/195) for Trypanosoma congolense (forest and savannah), 9.2% (18/195) for Trypanosoma brucei s.l. and 6.1% (12/195) for Trypanosoma vivax. The maps generated show the distribution of tsetse and trypanosome infections. This study showed an active transmission of trypanosomes in the wildlife reserve of Santchou. The maps enabled to identify areas with high transmission risk and where control operations must be implemented in order to eliminate tsetse and the diseases that they transmit.

Background: African trypanosomiases are caused by trypanosomes that are cyclically transmitted by tsetse. Investigations aiming to generate knowledge on the bacterial fauna of tsetse have revealed distinct symbiotic microorganisms. Furthermore, studies addressing the tripartite association between trypanosomes-tsetse-symbionts relationship have so far been contradictory. Most studies included Sodalis glossinudius and, consequently, the association involving Wolbachia is poorly understood. Understanding the vectorial competence of tsetse requires decrypting these tripartite associations. In this study, we identified Wolbachia and trypanosomes in Glossina palpalis palpalis from three human African trypanosomiasis (HAT) foci in southern Cameroon., Methods: Tsetse flies were captured with pyramidal traps in the Bipindi, Campo and Fontem HAT foci. After morphological identification, DNA was extracted from whole tsetse flies and Wolbachia and trypanosomes were identified by PCR using different trypanosome-specific primers and two Wolbachia-specific primers (Wolbachia surface protein and 16S rRNA genes). Statistical analyses were performed to compare the trypanosome and Wolbachia infection rates between villages and different foci and to look for an association between these microorganisms., Results: From a total of 2122 tsetse flies, 790 G. p. palpalis were analyzed. About 25.32% of flies hosted Wolbachia and 31.84% of non-teneral flies were infected by at least one trypanosome species. There was no significant difference between the global Wolbachia prevalence revealed by the two markers while some differences were observed between HAT foci. From 248 G. p. palpalis with trypanosome infections, 62.90% were with T. vivax, 34.68% with T. congolense forest, 16.13% with T. brucei (s.l.) and 2.42% with T. congolense savannah. Of all trypanosome-infected flies, 29.84% hosted Wolbachia and no association was observed between Wolbachia and trypanosome co-infections., Conclusions: This study revealed differences in the prevalence of Wolbachia and trypanosomes in G. p. palpalis according to HAT foci. The use of only one marker has underestimated the prevalence of Wolbachia, thus more markers in subsequent studies may improve its detection. The presence of Wolbachia seems to have no impact on the establishment of trypanosomes in G. p. palpalis. The tripartite association between tsetse, Wolbachia and trypanosomes varies according to studied areas. Studies aiming to evaluate the genetic polymorphism of Wolbachia and its density in tsetse flies could help to better understand this association.

Background: Changes of land cover modify the characteristics of habitat, host-vector interaction and consequently infection rates of disease causing agents. In this paper, we report variations in tsetse distribution patterns, abundance and infection rates in relation to habitat types and age in the Maasai Steppe of northern Tanzania. In Africa, Tsetse-transmitted trypanosomiasis negatively impacted human life where about 40 million people are at risk of contracting the disease with dramatic socio-economical consequences, for instance, loss of livestock, animal productivity, and manpower., Methods: We trapped tsetse flies in dry and wet seasons between October 2014 and May 2015 in selected habitats across four villages: Emboreet, Loiborsireet, Kimotorok and Oltukai adjacent to protected areas. Data collected include number and species of tsetse flies caught in baited traps, PCR identification of trypanosome species and extraction of monitored Normalized Difference Vegetation Index (NDVI) data from Moderate Resolution Imaging Spectrometer (MODIS)., Results: Our findings demonstrate the variation of tsetse fly species abundance and infection rates among habitats in surveyed villages in relation to NDVI and host abundance. Results have shown higher tsetse fly abundance in Acacia-swampy ecotone and riverine habitats for Emboreet and other villages, respectively. Tsetse abundance was inconsistent among habitats in different villages. Emboreet was highly infested with Glossina swynnertoni (68%) in ecotone and swampy habitats followed by G. morsitans (28%) and G. pallidipes (4%) in riverine habitat. In the remaining villages, the dominant tsetse fly species by 95% was G. pallidipes in all habitats. Trypanosoma vivax was the most prevalent species in all infected flies (95%) with few observations of co-infections (with T. congolense or T. brucei)., Conclusions: The findings of this study provide a framework to mapping hotspots of tsetse infestation and trypanosomiasis infection and enhance the communities to plan for effective control of trypanosomiasis.

Background: The control of animal trypanosomosis consists, amongst other things, of the punctual treatment of new cases, primarily diagnosed by pastoralists on the basis of clinical signs. This practice suggests that many apparently healthy infected animals are left untreated. In this study animal trypanosomosis in clinically healthy zebu cattle was evaluated, the distribution of the vectors established and the epidemiological implications discussed., Methods: In 2014 two cross-sectional surveys were carried out in the Cambeef ranch. A total of 866 blood samples were collected from cattle in different sites: 549 in the dry season and 317 in the rainy season. The blood samples were subjected to parasitological examination using the buffy coat method and to PCV determination. An entomological survey on animal trypanosomosis vectors was undertaken during tsetse flies caught were identified and the mid-gut of each living non-teneral tsetse fly was examined for infections using a microscope., Results: An overall trypanosomosis prevalence of 9% was found in the cattle examined. There were significantly (P < 0.05) more trypanosome infected cattle in the dry season than the rainy season. Trypanosome-infected cattle had significantly (P < 0.05) lower Body Condition Scores (BCS) and Packed Cell Volumes (PCV) in the dry season than in the rainy season. Anemia was positively correlated with trypanosome infection. The likelihood for an animal to be parasitologically free of trypanosome infection was at least three times as high in the Gudali breed as compared with the white and red Fulani breeds. Species of trypanosomes identified were Trypanosoma vivax (73.23%), Trypanosoma congolense (15.49%) and Trypanosoma brucei (11.27%). A total of 390 tsetse flies and 103 tabanids were trapped. Two species of tsetse flies were identified: Glossina tachinoides (33.59%) and G. morsitans submorsitans (41%). Nine of the 194 non-teneral flies were infected with trypanosomes., Conclusion: Carriers of trypanosomes are present amongst apparently healthy cattle in the study site. Attempts to successfully reduce the population of reservoir trypanosomes within herds and control the disease will need to consider mass screening once every year and this should be associated with drug sensitivity tests.

Background: The 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., Methods: Particular 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., Results: Conventional 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., Conclusion: Xenomonitoring 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.

Background: Fragmentation of tsetse habitat in eastern Zambia is largely due to encroachments by subsistence farmers into new areas in search of new agricultural land. The impact of habitat fragmentation on tsetse populations is not clearly understood. This study was aimed at establishing the impact of habitat fragmentation on physiological and demographic parameters of tsetse flies in order to enhance the understanding of the relationship between fragmentation and African animal trypanosomosis (AAT) risk., Methods: A longitudinal study was conducted to establish the age structure, abundance, proportion of females and trypanosome infection rate of Glossina morsitans morsitans Westwood (Diptera: Glossinidae) in areas of varying degrees of habitat fragmentation in Eastern Zambia. Black screen fly rounds were used to sample tsetse populations monthly for 1 year. Logistic regression was used to analyse age, proportion of females and infection rate data., Results: Flies got significantly older as fragmentation increased (p < 0.004). The proportion of old flies, i.e. above ovarian category four, increased significantly (P < 0.001) from 25.9% (CI 21.4-31.1) at the least fragmented site (Lusandwa) to 74.2% (CI 56.8-86.3) at the highly fragmented site (Chisulo). In the most fragmented area (Kasamanda), tsetse flies had almost disappeared. In the highly fragmented area a significantly higher trypanosome infection rate in tsetse (P < 0.001) than in areas with lower fragmentation was observed. Consequently a comparatively high trypanosomosis incidence rate in livestock was observed there despite lower tsetse density (p < 0.001). The overall proportion of captured female flies increased significantly (P < 0.005) as fragmentation reduced. The proportion increased from 0.135 (CI 0.10-0.18) to 0.285 (CI 0.26-0.31) at the highly and least fragmented sites, respectively., Conclusions: Habitat fragmentation creates conditions to which tsetse populations respond physiologically and demographically thereby affecting tsetse-trypanosome interactions and hence influencing trypanosomosis risk. Temperature rise due to fragmentation coupled with dominance of old flies in populations increases infection rate in tsetse and hence creates high risk of trypanosomosis in fragmented areas. Possibilities of how correlations between biological characteristics of populations and the degree of fragmentation can be used to structure populations based on their well-being, using integrated GIS and remote sensing techniques are discussed.

Background: Tsetse flies (Genus: Glossina) are the sole cyclical vectors of African trypanosomoses. Despite their economic and public health impacts in sub-Saharan Africa, it has been decades since the latest distribution maps at the continental level were produced. The Food and Agriculture Organization of the United Nations is trying to address this shortcoming through the Atlas of tsetse and African animal trypanosomosis., Methods: For the tsetse component of the Atlas, a geospatial database is being assembled which comprises information on the distribution and trypanosomal infection of Glossina species. Data are identified through a systematic literature review. Field data collected since January 1990 are included, with a focus on occurrence, apparent density and infection rates of tsetse flies. Mapping is carried out at the level of site/location. For tsetse distribution, the database includes such ancillary information items as survey period, trap type, attractant (if any), number of traps deployed in the site and the duration of trapping (in days). For tsetse infection, the sampling and diagnostic methods are also recorded., Results: As a proof of concept, tsetse distribution data for three pilot countries (Ethiopia, Kenya and Uganda) were compiled from 130 peer-reviewed publications, which enabled tsetse occurrence to be mapped in 1266 geographic locations. Maps were generated for eight tsetse species (i.e. G. brevipalpis, G. longipennis, G. fuscipes fuscipes, G. tachinoides, G. pallidipes, G. morsitans submorsitans, G. austeni and G. swynnertoni). For tsetse infection rates, data were identified in 25 papers, corresponding to 91 sites., Conclusions: A methodology was developed to assemble a geo-spatial database on the occurrence, apparent density and trypanosomal infection of Glossina species, which will enable continental maps to be generated. The methodology is suitable for broad brush mapping of all tsetse species of medical and veterinary public health importance. For a few tsetse species, especially those having limited economic importance and circumscribed geographic distribution (e.g. fusca group), recently published information is scanty or non-existent. Tsetse-infested countries can adopt and adapt this approach to compile national Atlases, which ought to draw also on the vast amount of unpublished information.

Background: Tsetse flies are the biological vectors of African trypanosomes, the causative agents of sleeping sickness in humans and nagana in animals. The tsetse endosymbiont Sodalis glossinidius has been suggested to play a role in tsetse susceptibility to infection. Here we investigate the prevalence of African trypanosomes within tsetse from the Luambe National Park, Zambia and if there is an association between S. glossinidius and presence of trypanosomes within the tsetse examined., Methods: Tsetse representing three species (Glossina brevipalpis, Glossina morsitans morsitans and Glossina pallidipes), were sampled from Luambe National Park, Zambia. Following DNA extraction, PCR was used to examine the tsetse for presence of trypanosomes and the secondary endosymbiont S. glossinidius., Results: S. glossinidius infection rates varied significantly between tsetse species, with G. brevipalpis (93.7%) showing the highest levels of infection followed by G. m. morsitans (17.5%) and G. pallidipes (1.4%). ITS-PCR detected a wide variety of trypanosomes within the tsetse that were analysed. Significant differences were found in terms of trypanosome presence between the three tsetse species. A high proportion of G. m. morsitans were shown to carry T. brucei s.l. DNA (73.7%) and of these around 50% were positive for Trypanosoma brucei rhodesiense. T. vivax, T. godfreyi, T. simiae, T. simiae Tsavo and T. congolense were also detected. No association was found between the occurrence of S. glossinidius and the presence of trypanosome DNA in any of the three tsetse species tested., Conclusion: The current work shows that T. b. rhodesiense was circulating in Luambe National Park, representing a risk for people living in the park or surrounding area and for tourists visiting the park. The differences in trypanosome DNA presence observed between the different tsetse species tested may indicate host feeding preferences, as the PCR will not discriminate between a fly with an active/resident infection compared to a refractory fly that has fed on an infected animal. This makes it difficult to establish if S. glossinidius may play a role in the susceptibility of tsetse flies to trypanosome infection.

Tsetse flies are the most important vectors of African trypanosomiasis but, surprisingly, are highly refractory to trypanosome parasite infection. In populations of wild caught flies, it is rare to find mature salivarian and mouthpart parasite infection rates exceeding 1 and 15%, respectively. This inherent refractoriness persists throughout the lifespan of the fly, although extreme starvation and suboptimal environmental conditions can cause a reversion to the susceptible phenotype. The teneral phenomenon is a phenotype unique to newly emerged, previously unfed tsetse, and is evidenced by a profound susceptibility to trypanosome infection. This susceptibility persists for only a few days post-emergence and decreases with fly age and bloodmeal acquisition. Researchers investigating trypanosome-tsetse interactions routinely exploit this phenomenon by using young, unfed (teneral) flies to naturally boost trypanosome establishment and maturation rates. A suite of factors may contribute, at least in part, to this unusual parasite permissive phenotype. These include the physical maturity of midgut barriers, the activation of immunoresponsive tissues and their effector molecules, and the role of the microflora within the midgut of the newly emerged fly. However, at present, the molecular mechanisms that underpin the teneral phenomenon still remain unknown. This review will provide a historical overview of the teneral phenomenon and will examine immune-related factors that influence, and may help us better understand, this unusual phenotype.

Background: Tsetse flies harbor at least three bacterial symbionts: Wigglesworthia glossinidia, Wolbachia pipientis and Sodalis glossinidius. Wigglesworthia and Sodalis reside in the gut in close association with trypanosomes and may influence establishment and development of midgut parasite infections. Wolbachia has been shown to induce reproductive effects in infected tsetse. This study was conducted to determine the prevalence of these endosymbionts in natural populations of G. austeni and G. pallidipes and to assess the degree of concurrent infections with trypanosomes., Methods: Fly samples analyzed originated from Kenyan coastal forests (trapped in 2009-2011) and South African G. austeni collected in 2008. The age structure was estimated by standard methods. G. austeni (n=298) and G. pallidipes (n= 302) were analyzed for infection with Wolbachia and Sodalis using PCR. Trypanosome infection was determined either by microscopic examination of dissected organs or by PCR amplification., Results: Overall we observed that G. pallidipes females had a longer lifespan (70 d) than G. austeni (54 d) in natural populations. Wolbachia infections were present in all G. austeni flies analysed, while in contrast, this symbiont was absent from G. pallidipes. The density of Wolbachia infections in the Kenyan G. austeni population was higher than that observed in South African flies. The infection prevalence of Sodalis ranged from 3.7% in G. austeni to about 16% in G. pallidipes. Microscopic examination of midguts revealed an overall trypanosome infection prevalence of 6% (n = 235) and 5% (n = 552), while evaluation with ITS1 primers indicated a prevalence of about 13% (n = 296) and 10% (n = 302) in G. austeni and G. pallidipes, respectively. The majority of infections (46%) were with T. congolense. Co-infection with all three organisms was observed at 1% and 3.3% in G. austeni and G. pallidipes, respectively. Eleven out of the thirteen (85%) co-infected flies harboured T. congolense and T. simiae parasites. While the association between trypanosomes and Sodalis infection was statistically significant in G. pallidipes (P = 0.0127), the number of co-infected flies was too few for a definite conclusion., Conclusions: The tsetse populations analyzed differed in the prevalence of symbionts, despite being sympatric and therefore exposed to identical environmental factors. The density of infections with Wolbachia also differed between G. austeni populations. There were too few natural co-infections detected with the Sodalis and trypanosomes to suggest extensive inter-relations between these infections in natural populations. We discuss these findings in the context of potential symbiont-mediated control interventions to reduce parasite infections and/or fly populations.

Insect symbioses lack the complexity and diversity of those associated with higher eukaryotic hosts. Symbiotic microbiomes are beneficial to their insect hosts in many ways, including dietary supplementation, tolerance to environmental perturbations and maintenance and/or enhancement of host immune system homeostasis. Recent studies have also highlighted the importance of the microbiome in the context of host pathogen transmission processes. Here we provide an overview of the relationship between insect disease vectors, such as tsetse flies and mosquitoes, and their associated microbiome. Several mechanisms are discussed through which symbiotic microbes can influence the ability of their host to transmit pathogens, as well as potential disease control strategies that harness symbiotic microbes to reduce pathogen transmission through an insect vector., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

In the Mouhoun River basin, Burkina Faso, the main vectors of African animal trypanosomoses are Glossina palpalis gambiensis Vanderplank and Glossina tachinoides Westwood (Diptera: Glossinidae), both of which are riverine tsetse species. The aim of our study was to understand the impact of landscape anthropogenic changes on the seasonal dynamics of vectors and associated trypanosomosis risk. Three sites were selected on the basis of the level of disturbance of tsetse habitats and predominant tsetse species: disturbed (Boromo, for G. tachinoides) and half-disturbed (Douroula for G. tachinoides and Kadomba for G. p. gambiensis). At each of these sites, seasonal variations in the apparent densities of tsetse and mechanical vectors and tsetse infection rates were monitored over 17 months. Tsetse densities differed significantly between sites and seasons. Of 5613 captured tsetse, 1897 were dissected; 34 of these were found to be infected with trypanosomes. The most frequent infection was Trypanosoma vivax (1.4%), followed by Trypanosoma congolense (0.3%) and Trypanosoma brucei (0.05%). The mean physiological age of 703 tsetse females was investigated to better characterize the transmission risk. Despite the environmental changes, it appeared that tsetse lived long enough to transmit trypanosomes, especially in half-disturbed landscapes. A total of 3021 other biting flies from 15 species (mainly Tabanidae and Stomoxyinae) were also caught: their densities also differed significantly among sites and seasons. Their relative importance regarding trypanosome transmission is discussed; the trypanosomosis risk in cattle was similar at all sites despite very low tsetse densities (but high mechanical vector densities) in one of them.

The blood meal origins of 222 tsetse flies (213 Glossina palpalis palpalis, 7 Glossina pallicera pallicera, one Glossina nigrofusca and one Glossina caliginea) caught in 2008 in two Human African trypanosomiasis foci (Bipindi and Campo) of south Cameroon were investigated. 88.7% of tsetse flies blood meals were identified using the heteroduplex method and the origin of the remaining blood meals (11.3%) was identified by sequencing the cytochrome B gene. Most of the meals were from humans (45.9%) and pigs (37.4%), 16.7% from wild animals. Interestingly, new tsetse fly hosts including turtle (Trionyx and Kinixys) and snake (Python sebae) were identified. Significant differences were recorded between Bipindi where the blood meals from pigs were predominant (66.7% vs 23.5% from humans) and Campo where blood meals from humans were predominant (62.9% vs 22.7% from pigs). Comparison with the data recorded in 2004 in the same foci (and with the same molecular approach) demonstrated significant modifications of the feeding patterns: increase in blood meals from pigs in Bipindi (66.7% in 2008 vs 44.8% in 2004) and in Campo (20.5% in 2008 vs 6.8% in 2004), decrease in that from human (significant in Bipindi only). 12.6%, 8.1% and 2.7% of the flies were, respectively, Trypanosoma congolense forest type, Trypanosoma congolense savannah type and Trypanosoma brucei gambiense infected. These results demonstrate that tsetse fly feeding patterns can be specific of a given area and can evolve rapidly with time. They show an active circulation of a variety of trypanosomes in sleeping sickness foci of southern Cameroon., (Copyright 2010 Elsevier B.V. All rights reserved.)

In an on-farm trial conducted amongst the Maasai pastoralists in Nkuruman and Nkineji areas of Kenya between April 2004 and August 2005 designed to evaluate the effectiveness of a synthetic tsetse repellent technology, we assessed the relationship between tsetse challenge and trypanosomosis incidence in cattle. Six villages were used in each area. Each of these villages had a sentinel cattle herd that was screened for trypanosomosis on monthly basis using buffy coat technique. Animals found infected at each sampling were treated with diminazene aceturate at 7 mg kg(-1) body weight. Treatments administered by the owners over the sampling intervals were recorded as well. Tsetse flies were trapped at the time of sampling using baited stationary traps and apparent tsetse density estimated as flies per trap per day (FTD). A fixed proportion (10%) of the flies was dissected and their infection status determined through microscopy. Blood meals were also collected from some of the flies and their sources identified using enzyme-linked immunosorbent assay (ELISA). Tsetse challenge was obtained as a product of tsetse density, trypanosome prevalence and the proportion of blood meals obtained from cattle. This variable was transformed using logarithmic function and fitted as an independent factor in a Poisson model that had trypanosomosis incidence in the sentinel cattle as the outcome of interest. The mean trypanosomosis incidence in the sentinel group of cattle was 7.2 and 10.2% in Nkuruman and Nkineji, respectively. Glossina pallidipes was the most prevalent tsetse species in Nkuruman while G. swynnertoni was prevalent in Nkineji. The proportions of tsetse that had mature infections in the respective areas were 0.6 and 4.2%. Most tsetse (28%) sampled in Nkuruman had blood meals from warthogs while most of those sampled in Nkineji (30%) had blood meals from cattle. A statistically significant association between tsetse challenge and trypanosomosis incidence was obtained only in Nkuruman when data was pooled and analyzed at the area but not at the village-level. In the later scenario, lagging tsetse challenge by 1 month improved the strength but not the significance of the association. These findings show that when the spatial unit of analysis in observational studies or on-farm trials is small, for instance a village, it may not be possible to demonstrate a statistically significant association between tsetse challenge and trypanosomosis incidence in livestock so as to effectively control for tsetse challenge.

Trypanosomal infections in female Glossina morsitans morsitans were investigated in an area in the Eastern Province of Zambia between 1992 and 1994. A total of 4416 flies were captured, aged using the ovarian ageing method and screened for trypanosomal infections in both the mouthparts, salivary glands and the midgut. Congolense-type infections were identified in 4.8% of the flies. Vivax-type and immature infections were identified in 1.8% and 6.8% of the flies, respectively. The prevalence of congolense-type, vivax-type and immature infections increased with age. For vivax-type infections the age-prevalence relationship could be described by a model assuming a constant per capita rate of infection. For congolense-type and midgut infections, a polynomial term was added to the model significantly improving the fit. The per capita at which flies become infected was significantly higher for immature compared to mature infections. Observations strongly suggest that tsetse acquire new midgut infections at any age and that maturation of these infections is not limited to those obtained during the first blood meal.

TSETSE FLIES (DIPTERA: Glossinidae) are important agricultural and medical vectors transmitting the African trypanosomes, the agents of sleeping sickness disease in humans and various diseases in animals (nagana). While the prevalence of disease has increased to epidemic proportions, lack of a mammalian vaccine and affordable and effective drugs have hindered disease control. Trypanosomiasis management relies heavily on the control of its single insect vector, the tsetse fly. Despite the effectiveness of some of these tools, their impact on disease control has not been sustainable due to their local nature and extensive dependence on community participation. Recent advances in molecular technologies and their application to insects have revolutionized the field of vector biology, and there is hope that such new approaches may form the basis for future tsetse interventions. The success of the genetic approaches aiming to disrupt the transmission cycle of the parasite in their invertebrate host depends on full understanding of the interaction between tsetse and trypanosomes. This article reviews the biology of trypanosome development in the fly and the multiple bacterial symbionts that inhabit the same gut environment. The availability of a genetic transformation system for the midgut symbiont allows for gene products to be expressed in vivo in the tsetse gut where they can produce a hostile environment for pathogen transmission. The characterization of gene product(s) with anti-pathogenic properties and their expression in vivo is discussed. A strategy is outlined where the replacement of susceptible insect phenotypes with their engineered refractory counterparts can result in decreased disease transmission.

Tsetse flies (Diptera: Glossinidae) are vectors of several species of pathogenic trypanosomes in tropical Africa. Human African trypanosomiasis (HAT) is a zoonosis caused by Trypanosoma brucei rhodesiense in East Africa and T. b. gambiense in West and Central Africa. About 100000 new cases are reported per year, with many more probably remaining undetected. Sixty million people living in 36 countries are at risk of infection. Recently, T. b. gambiense trypanosomiasis has emerged as a major public health problem in Central Africa, especially in the Democratic Republic of Congo, Angola and southern Sudan where civil war has hampered control efforts. African trypanosomes also cause nagana in livestock. T. vivax and T. congolense are major pathogens of cattle and other ruminants, while T. simiae causes high mortality in domestic pigs; T. brucei affects all livestock, with particularly severe effects in equines and dogs. Central to the control of these diseases is control of the tsetse vector, which should be very effective since trypanosomes rely on this single insect for transmission. However, the area infested by tsetse has increased in the past century. Recent advances in molecular technologies and their application to insects have revolutionized the field of vector biology, and there is hope that such new approaches may form the basis for future tsetse control strategies. This article reviews the known biology of trypanosome development in the fly in the context of the physiology of the digestive system and interactions of the immune defences and symbiotic flora.

No sleeping sickness or nagana cases have been reported in Botswana since 1985. In view of several confirmed clinical cases of nagana and reports of heavy bovine mortality, a parasitological survey was conducted to determine the prevalence of trypanosome infection in cattle in Maun and Shakawe areas of Ngamiland district. Wet blood films, buffy coat and Giemsa-stained thick and thin blood smears were used to detect trypanosomes in animals. Overall,trypanosome infection rate was 15.98%, with 5.94% and 27.29% in Maun and Shakawe respectively. The urgent need to combat trypanosomosis in Ngamiland, particularly in the Shakawe area, is highlighted, and a 3-phase integrated tsetse control strategy for this disease problem is discussed.

The polymerase chain reaction was used to characterize the trypanosomes infecting Glossina morsitans submorsitans and G. tachinoides in the game ranch of Nazinga, Burkina Faso, situated near an agropastoral zone. Dissection of 435 tsetse flies, and PCR analysis of 166 infected flies were conducted to assess the epidemiological situation. Trypanosomes of the Nannomonas subgenus were the most abundant in the two tsetse species (80.4% and 73.7% of identified infections in G. m. submorsitans and G. tachinoides respectively). T. vivax and T. brucei infection rates were comparable between the two tsetse species. Mature infection pattern identified by PCR differed from overall infections, mainly because T. simiae infections did not mature, whereas T. vivax represented the predominant taxon. Parasitological and PCR results showed some discrepancies; possibly some typical Duttonella strains could not be recognized by the sets of primers used. The technologies used in this work helped to determine the high trypanosomosis risk in this area.

Trypanotolerance reaches varying degrees of stability in wild and domestic animals according to their co-evolutionary contacts with tsetse flies and trypanosomes. In this context, various mechanisms developed by wild and domestic Bovidae to cope with tsetse and trypanosome challenge are discussed.

Over recent years there has been an increasing interest in the interaction between Trypanosomatids and in particular the genera Trypanosoma and Leishmania and their vectors. These studies on the "vector parasite interface" have been wide ranging but often suffer from the difficulties of manipulating some of the vector systems. It is now likely that molecular biological and protein analysis techniques can be more effectively applied in dissecting these interactions with a view to identification and characterisation of molecules which control parasite behaviour in vectors. This presentation will review recent studies on Glossina lectins in relation to trypanosome behaviour in flies including recent studies on different G. palpalis subspecies as well as review evidence that trypanosomatids have an effect on their insect hosts for example, on feeding behaviour, susceptibility to insecticides and longevity of infected vectors. Sugars are known to influence development and transmission of Leishmania and recent studies have indicated that aphid/or coccid honeydew is taken by sandflies, the relevance of this to Leishmania/sandfly interactions will be discussed. The basic mechanisms of attachment observed in all trypanosomatid interactions are described and the presence of molecules associated with attachment have been identified. The genetic basis of susceptibility is also becoming better understood and it is to be expected that modern molecular techniques when applied to well defined systems can give results which could permit an attempt at intervention; even if this is not achieved the basic understanding of a widespread phenomenon of the insect/parasite association will have been furthered to permit a better epidemiological knowledge.

Tenerals of Glossina morsitans morsitans and G. m. centralis were infected with Trypanosoma vivax, T. congolense or T. brucei by feeding mainly on infected goats and then maintained either in vivo on uninfected calves, goats or rabbits, or fed in vitro upon heparinised or defibrinated blood of goats or calves for 21 days for T. vivax and T. congolense and 30 days for T. brucei and then dissected. The observed differences in the infection rates for all three trypanosome species maintained on different diets were small and/or inconsistent and possibly are of no significance. It is therefore likely that the in vitro feeding of the tsetse on these diets after infected blood meal has no adverse effect on the cyclical development of these trypanosome species in these vectors.

Genetic exchange may occur between two particular Trypanosoma brucei clones simultaneously transmitted by the same tsetse fly. We report here that this exchange takes place in the fly, through nuclear fusion. The resulting hybrids appear to be sub-tetraploid, some particular DNA sequences from one of the parental stocks being lost before enough cloned hybrid trypanosomes could be harvested for DNA analysis. A further reduction of the DNA content of these hybrids occurs gradually upon growth and yields near diploid value in a major part of the population. This mode of hybrid generation is different from the fusion of haploid gametes, which is thought to occur normally upon inoculation of metacyclic trypanosomes in their mammalian host. In this respect, the sub-tetraploid hybrids appear to undergo meiosis in the fly, generating sub-diploid metacyclic forms, then fusion in the mammalian blood.

The fluid mechanics of the flow of blood through the labrum of Glossina is described in uninfected and trypanosome infected flies. The flow is characterised by the Reynolds number, and a frequency parameter for the pulsating flow and non-newtonian viscosity effects are considered. The effects of colonies of trypanosomes on the flow rate in the labrum and the interactions between colonies of Trypanosoma (Nannomonas) congolense and Trypanosoma (Trypanozoon) brucei and mechanoreceptors in the proximal third of the labrum are described. The direct association between trypanosomes and mechanoreceptors and the reduction in flow rate imposed by trypanosome colonies in the food canal distal to the mechanoreceptors will impair detection of stimuli by them. The epidemiological implications of these and earlier observations are discussed.

Using biconical traps the distribution, population composition, insemination rate, pregnancy, age structure and trypanosome infection rate of G. pallidipes were studied during the wet season 1982 and the dry season 1983 at Mareerrey Somalia. Flies restricted to the riverine gallery forest in the dry season become dispersed into approximately 1 km of the Acacia thickets in the wet season. There was no significant variation in population components of G. pallidipes between wet and dry seasons. The male:female ratio remained at 0.5:1 in both seasons. The insemination rate of females was significantly high during the wet season. Pregnancy stages were not statistically different between the two study periods. A significantly greater proportion of females that were below age category 4 were found in the wet season. The trypanosome infection rate was 2.6% and 1.5% in the wet and dry seasons respectively. Flies were found infected with only the vivax and congolense group trypanosomes. A linear positive correlation existed between the trypanosome infection rate and the physiological age of females.

The feeding behaviour of Glossina morsitans morsitans infected with Trypanosoma vivax, T. congolense or T. brucei was studied and compared with that of uninfected control tsetse. The following parameters were measured. (1) Mean number of feeds taken in 20 consecutive days; (2) mean rate of blood meal intake in microgram/sec of the total days fed; (3) mean weight of blood meal intake/day of the total days fed; (4) mean total blood meal engorgement in 20 days; (5) mean number of probes/day of the days fed; (6) mean frequency of the multiple probe feeds and (7) mean number of pre-feeding probes of the multiple probe feeds. Evidence was found to suggest that the rosettes of T. vivax and T. congolense in the proboscis, and colonies of T. brucei parasites in the salivary glands, did not significantly alter the feeding behaviour of the vector.

A population of Glossina longipalpis studied in the Southern Guinea savanna zone, Ivory Coast, showed marked differences between non-infected and infected females. Each fly was examined for age, reproductive condition, wear and tear, size and nutritional status. Infected flies were generally and sometimes significantly more active with lower fat reserves, residual bloodmeal and higher residual dry weight. Poorer nutritional condition may be due to energy metabolised by trypanosomes and possibly impaired feeding ability.