A phyto-sociological analysis of the distribution of riverine tsetse flies in Burkina Faso.

In Burkina Faso, Glossina palpalis gambiensis Vanderplank and G. tachinoides Westwood (Diptera: Glossinidae) are the main cyclic vectors of trypanosomiasis. The vegetation type along river banks is an important factor determining the distribution and abundance of these tsetse. The following work investigated the relation between the plant species present (including the disturbance level) and tsetse distribution and abundance, using three ecotypes, described by P.C. Morel in 1978. These were the Guinean, Sudano-Guinean and Sudanese gallery forests. In the Mouhoun River basin, these three ecotypes are found successively from upstream to downstream. Berlinia grandiflora, Syzygium guineense and Cola laurifolia and finally Acacia seyal and Mitragyna inermis were the best indicators for the Guinean, Sudano-Guinean and Sudanese gallery forest ecotypes, respectively, as suggested by Morel. However, other species such as Pterocarpus santalinoides and Mimosa pigra were not ecotype specific. Trap catches confirmed that G. palpalis and G. tachinoides are predominant in Guinean and Sudanese gallery forests, respectively, and that both species are well represented in the Sudano-Guinean ecotype. Tsetse densities dropped significantly in disturbed Sudano-Guinean and Sudanese gallery forest sites. However, this was not the case for both species in Guinean or for G. tachinoides in half-disturbed Sudanese gallery forest sites, confirming their high resilience to human-made changes. The importance of a detailed consideration of riverine ecotypes when predicting tsetse densities is discussed.

Geographical Information Systems in parasitology: a review of potential applications using the example of animal trypanosomosis in West Africa.

The epidemiology of vector-borne diseases is complex due to the variability in the ecology of the different actors involved, i.e. hosts, parasites and vectors. The transmission of African animal trypanosomosis in the West-African savannah region is an excellent example of this complexity: riverine tsetse flies have an heterogeneous distribution along the rivers, depending of suitable habitats, and transmit pathogenic trypanosomes were they use domestic animal as feeding hosts. Contrasting epidemiological situations may thus occur at the local scale, and a broad view of the overall environment is necessary to quantify the interfaces in time and space between hosts and vectors. Geographical Information Systems (GIS) can provide new insight into the study of such complex epidemiological processes. GIS is a powerful technology that has been used mainly in map-making, and an enormous amount of knowledge can be gained simply by geographical data projection. GIS also allows juxtaposition of different types of information, creation of new variables, testing of theories and correlation, and generating of predictive models. The purpose of the present paper is to exemplify the potential application of GIS using a recent study carried out on animal trypanosomosis in a cattle-raising area of Burkina Faso.

Inhibition of the DNA amplification of trypanosomes present in tsetse flies midguts: implications for the identification of trypanosome species in wild tsetse flies.

The present study was carried out in order to investigate if there was really a failure of PCR in identifying parasitologically positive tsetse flies in the field. Tsetse flies (Glossina palpalis gambiensis and Glossina morsitans morsitans) were therefore experimentally infected with two different species of Trypanosoma (Trypanosoma brucei gambiense or Trypanosoma congolense). A total of 152 tsetse flies were dissected, and organs of each fly (midgut, proboscis or salivary glands) were examined. The positive organs were then analysed using PCR. Results showed that, regardless of the trypanosome species, PCR failed to amplify 40% of the parasitologically positive midguts. This failure, which does not occur with diluted samples, is likely to be caused by an inhibition of the amplification reaction. This finding has important implications for the detection and the identification of trypanosome species in wild tsetse flies.

Monitoring the susceptibility of Glossina palpalis gambiensis and G. morsitans morsitans to experimental infection with savannah-type Trypanosoma congolense, using the polymerase chain reaction.

Teneral Glossina palpalis gambiensis and G. morsitans morsitans (Diptera: Glossinidae) were fed on mice infected with savannah-type Trypanosoma (Nannomonas) congolense. The infection was monitored by checking the post-feeding diuresis fluid (midgut infection) and saliva (mature infection) of individual flies for parasites, at different times post-infection, using microscopical examination and a PCR-based assay. The results indicated that both tsetse species supported established midgut infections by 10 days post-infection and that maturation occurred after 24 days in G. m. morsitans. Although, for both diuresis fluid and saliva, the results of the microscopy showed good concordance with those of the PCR, the PCR identified more positive samples. Monitoring allowed determination of the status of the infection in individual flies, which was confirmed, 48 days post-infection, by the microscopical examination of the midguts and probosces dissected out of the flies and by the PCR-based amplification of any trypanosome DNA in these organs. Again, in terms of the detection of trypanosomes in the dissected organs, there was good concordance between the results of the PCR and those of the microscopy, although PCR revealed many more mature infections than did microscopical examination, particularly in the G. p. gambiensis investigated. There was a higher prevalence of immature infection in G. p. gambiensis than in G. m. morsitans (P<0.05) but the inter-specific differences seen in the prevalences of any infection and of mature infection were not statistically significant. The intrinsic vectorial capacity for T. congolense of both tsetse species therefore appeared quite similar, although the true vectorial competence of G. p. gambiensis remains to be determined.

Monitoring the developmental status of Trypanosoma brucei gambiense in the tsetse fly by means of PCR analysis of anal and saliva drops.

Teneral Glossina palpalis gambiensis (Diptera: Glossinidae) were infected with a culture of procyclic forms of Trypanosoma brucei gambiense using a single-bloodmeal membrane feeding technique. The infection was monitored by analysing the saliva (mature infection) and anal drop (midgut infection) of each fly at different post-infection times both by microscopic observation and polymerase chain reaction (PCR). Amplification revealed many more positive anal drops than microscopy. The monitoring showed that the installation of T. b. gambiense in Glossina took place at least 11 days after the infection and that maturation occurred after 29 days. It also reflected precisely the parasitic status of each tsetse fly as determined by the dissection, microscopic examination and PCR amplification of the midguts and salivary glands 47 days post-infection. Twice as many tsetse flies with mature salivary glands infection were revealed by PCR than by microscopic examination, but the two techniques gave exactly the same results regarding the proportion of flies with midgut infection. This study also demonstrated the ability of natural non-infective procyclic forms of T. b. gambiense, to colonise the midgut and subsequently establish in the salivary glands of G. p. gambiensis.

[Tsetse fly wings, an identity card of the insect?]. / Les ailes de glossines, une carte d'identité de l'insecte?

The size of tsetse flies is often associated with population dynamics and vectorial capacity parameters. Adult fly size is generally estimated from measurements of wing segments. To take measure of the wing, a semi-automatic software was developed by CIRAD-EMVT and IRD. It was used in wild populations of Glossina tachinoides Westwood and G. palpalis gambiensis Vanderplank (Diptera: Glossinidae) trapped near Bobo-Dioulasso, Burkina Faso. From an numeric picture of the wing, the software calculates the length of vein segments, the ratios between these lengths, the surface of the tsetse characteristic "hatchet cell", and the greyness on the wings. The data were interesting at the level of taxonomy. In addition, they help specify physiological characteristics of the studied populations.

[New tools for the study of animal trypanosomiasis in the Sudan: model-building of dangerous epidemiological passage by remote sensing geographic information systems]. / De nouveaux outils pour l'étude des trypanosomoses animales en zone soudanienne: modélisation de paysages épidémiologiquement dangereux par télédétection et Systèmes d'Information Géographique.

Recent studies in a rangeland area of Burkina Faso showed that riparian tsetse flies (Glossina tachinoides and G. palpalis gambiensis) were found along the main rivers, but depending on their location, they had different hosts and were not infected by the same trypanosomoses. There were different epidemiological situations within a distance of a few kilometres, and local assessment of the trypanosome risk thus called for a global approach taking account of the environmental and human factors involved in the interfaces between hosts and vectors. Various types of information concerning entomology, parasitology, ecology, land occupation and animal production systems were fed into a Geographical Information System. High spatial resolution remote sensing tools and original modelling methods were used to detect the valley landscapes most favourable to tsetse flies, and describe land use by herds. The impact of trypanosomes appeared to depend largely on animal movements, watering practices and the degree of contact with riparian tsetse flies. Linking these types of information revealed the most dangerous sites in epidemiological terms, which in this case represented some 18% of the network initially surveyed.

The changing distribution of two riverine tsetse flies over 15 years in an increasingly cultivated area of Burkina Faso.

Changes in the distribution of two riverine tsetse flies, Glossina tachinoides Westwood and Glossina palpalis gambiensis Vanderplank are described in an agro-pastoral area of Burkina Faso subject to increasing human population pressure and land use change. Two similar entomological surveys (one trap every 100 m, 120 km of river) were conducted in 1981 and 1996. Changes in tsetse distribution were compared to land use changes through high resolution remote sensing imagery (LANDSAT, SPOT). There was a close relationship between proximity of crops relative to riverine forest and the density of Glossina. Where fields encroached on riverine vegetation, tsetse populations declined. Where the geomorphological structure was not well suited to agricultural activity, riverine vegetation and tsetse fly populations were relatively unaffected, even with intense agricultural activity nearby. In contrast, increased human activity and higher cattle densities in the surrounding savannah areas were associated with increased tsetse numbers. The results demonstrated a wide diversity of tsetse distribution and habitat within a few kilometres in an agro-pastoral landscape in West Africa.

[Different potentials of a geographic information system for studies in epidemiology: example of animal trypanosomiasis in the Sudan region]. / Les différentes potentialités d'un système d'information géographique pour les études en épidemiologie: l'exemple des trypanosomoses animales en zone soudanienne.

The epidemiology of vector-borne diseases is complex due to variability in the ecology of the different actors involved, i.e., hosts, parasites and vectors. Transmission of animal trypanosomiasis in the West African savannah region is an excellent example of this complexity. Because of the importance of place and time in determining the interface between host and vector, a broad view of the overall environment is necessary to understand the variety of situations that can be encountered in an epidemiological system. Spatial distribution is a key factor for risk evaluation. State-of-the-art geographic information systems (GIS) have provided new insight into this complex epidemiology. GIS is a powerful technology that has been used mainly in map-making. An enormous amount of knowledge can be gained simply by geographical data projection. However GIS also allows juxtaposition of different types of information, creation of new variables, testing of theories and correlations, and generation of predictive models. The purpose of this article is to exemplify the potential applications of GIS using a recent study carried out on animal trypanosomiasis in a cattle-raising area of Burkina Faso.

Microsatellite DNA markers reveal genetic differentiation among populations of Glossina palpalis gambiensis collected in the agro-pastoral zone of Sideradougou, Burkina Faso.

Intraspecific genetic variability of Glossina palpalis gambiensis in the area of Sideradougou, Burkina Faso, was studied using polymorphic microsatellite DNA markers. This genetic study was combined with other epidemiological information on the same tsetse: bloodmeal identification, dissection of tsetse and molecular characterization of the trypanosomes detected. There was significant genetic differentiation among flies caught only a few kilometers apart, within the same riverine habitat. These distinct subpopulations were also differentially infected by trypanosomes. In part of the study area, a Factorial Correspondence Analysis undertaken on the genotypes allowed us to detect a Wahlund effect, suggesting the presence of tsetse originating from different source populations coming from two distinct drainage systems. The apparent structuring of populations of G. palpalis gambiensis is discussed relative to appropriate strategies to control African Trypanosomosis.

Intraspecific variability in natural populations of Glossina palpalis gambiensis from West Africa, revealed by genetic and morphometric analyses.

Glossina palpalis gambiensis Vanderplank (Diptera: Glossinidae) from West Africa (Senegal and Burkina Faso) were analysed for microsatellite DNA polymorphisms and size of the wings. In the overall sample a strong heterozygote deficiency was found at two polymorphic microsatellite loci. It led to a highly significant value of Fis (within-sample heterozygote deficit) in the western zone of Sideradougou area in Burkina Faso. Genetic differentiation was significant on a macrogeographic scale, i.e. between tsetse coming from Senegal and Burkina Faso. Wing measures also differed between these two countries; flies from Senegal appeared to be smaller. Microsatellite loci further allowed differentiation of populations of G. palpalis gambiensis trapped on the same hydrographic network a few kilometres apart. The results are interpreted as indicating that further investigations will allow the study of genetic variability of tsetse flies in relation to the dynamics of transmission of human and animal trypanosomoses.

Polymerase chain reaction as a diagnosis tool for detecting trypanosomes in naturally infected cattle in Burkina Faso.

African animal trypanosomoses constitute the most important vector-borne cattle diseases in sub-Saharan Africa. Generally it is considered that there is a great lack of accurate tools for the diagnosis of the disease. During a trypanosomosis survey in the agro-pastoral zone of Sideradougou, Burkina Faso, 1036 cattle were examined for trypanosomes using microscopy. The PCR was applied on a subset of 260 buffy-coat samples using primers specific for Trypanosoma congolense savannah and riverine-forest groups, T. vivax, and T. brucei. Parasitological examination and the molecular technique were compared, showing a better efficiency of the latter. In the near future, the PCR is likely to become an efficient tool to estimate the prevalence of African trypanosomoses in affected areas.

Review on the molecular tools for the understanding of the epidemiology of animal trypanosomosis in West Africa.

The epidemiology of animal trypanosomosis around Bobo-Dioulasso (Burkina Faso, West Africa) benefited a lot in the last years from the progress of molecular tools. The two most used molecular techniques were the polymerase chain reaction for the diagnosis of the disease in cattle and the characterization of the trypanosomes in the host and the vector on one hand, and the microsatellite DNA polymorphism in tsetse flies to study the intraspecific genetic variability of the vector on the other hand. The results obtained in the Sideradougou area during a recent two year survey with these techniques, associated with many other georeferenced informations concerning vector and cattle distribution, natural environment, landuse, ground occupation, livestock management, were combined in a Geographical Information System. This new approach of a complex pathogenic system led to a better evaluation of the risk of trypanosome transmission.

Microsatellite markers for genetic population studies in Glossina palpalis gambiensis (Diptera: Glossinidae).

Little is known about intraspecific variability in tsetse flies and its consequences for vectorial capacity. Microsatellite markers have been developed for Glossina palpalis gambiensis. Three loci have been identified and showed size polymorphisms for insectarium samples. G. palpalis gambiensis from Burkina Faso were also subjected to PCR to investigate then genetic variability. Amplifications were observed in different species belonging to the palpalis group. These molecular markers will be useful to estimate gene flow within G. palpalis gambiensis populations and analysis could be extended to related species.

New epidemiological features on animal trypanosomiasis by molecular analysis in the pastoral zone of Sideradougou, Burkina Faso.

A multidisciplinary work was undertaken in the agropastoral zone of Sidéradougou, Burkina Faso to try to elucidate the key factors determining the presence of tsetse flies. In this study the PCR was used to characterize trypanosomes infecting the vector (Glossina tachinoides and Glossina palpalis gambiensis) and the host, i.e. cattle. A 2-year survey involved dissecting 2211 tsetse of the two Glossina species. A total of 298 parasitologically infected tsetse were analysed by PCR. Trypanosoma vivax was the most frequently identified trypanosome followed by the savannah type of T. congolense and, to a lesser extent, the riverine forest type of T. congolense, and by T brucei. No cases of T. simiae were found. From the 107 identified infections in cattle, the taxa were the same, but T. congolense savannah type was more frequent, whereas T. vivax and T. congolense riverine forest types were found less frequently. A correlation was found between midgut infection rates of tsetse, nonidentified infections and reptile bloodmeals. These rates were higher in G.p. gambiensis, and in the western part of the study area. T. vivax infections were related to cattle bloodmeals, and were more frequent in G. tachinoides and in the eastern study area. The PCR results combined with bloodmeal analysis helped us to establish the relationships between the vector and the host, to assess the trypanosome challenge in the two parts of the area, to elucidate the differences between the two types of T. congolense, and to suspect that most midgut infections were originating from reptilian trypanosomes.

PCR analysis and spatial repartition of trypanosomes infecting tsetse flies in Sidéradougou area of Burkina Faso.

A parasitological and entomological survey was conducted in the Sideradougou area (south of Bobo Dioulasso, Burkina Faso) in order to identify transmission factors of African trypanosomosis. A total of 3600 tsetse flies (Glossina tachinoides, Glossina palpalis gambiensis) were captured along 120 km of linear gallery forest and half of them were dissected. PCR analysis was undertaken on parasitologically positive flies (161 G. tachinoides, 92 G. palpalis gambiensis) to characterize the different trypanosomes. All the results were integrated in a GIS (Geographical Information System). Spatial repartition of the characterized trypanosomes enabled to recognize different areas with specific patterns of infection. Relations with environmental factors are discussed.

Molecular characterization of trypanosome isolates from naturally infected domestic animals in Burkina, Faso.

A total of 33 trypanosome cryostabilates isolated from domestic animals (bovine and dogs) were analysed using the polymerase chain reaction (PCR). The PCR was undertaken on diluted and treated buffy coat solutions according to an easy protocol of purification, using primers specific to Trypanosoma (Nannomonas) congolense of Savannah, Riverine-Forest, Kilifi and Tsavo types, T. (N) simiae, T. (Trypanozoon) brucei and T. (Duttonella) vivax. The results showed a lack of PCR sensitivity when target solutions were simply diluted, probably a reflection of the inaccuracy of the dilution procedure at very low trypanosome numbers. Nine mixed infections were found in purified samples whereas only three were detected in diluted crude solutions. T. congolense Savannah-type was present in all stabilates. Double infections involving this type with the Riverine-Forest type, T. vivax or T. brucei, were found. One stabilate was found to be infected with the three trypanosome types, namely T. congolense Savannah and Riverine-Forest genotypes and T. vivax. No infection attributable to T. congolense Kilifi and Tsavo types or T. simiae was detected in these stabilates. This work confirmed the abundance of mixed infections in the field, which could not have been detected by the classical parasitological methods. Amongst the T. congolense infections, the Savannah genotype was found to be predominant over the Riverine-Forest type; that could be a consequence of differences in genotype virulence in cattle. The detection of T. congolense Riverine-Forest type in vertebrate hosts living in wet areas could be confirmation of the suspected affinity of relationships between this taxa and the riverine forest tsetse fly species.

Comparison of the susceptibility of different Glossina species to simple and mixed infections with Trypanosoma (Nannomonas) congolense savannah and riverine forest types.

Teneral Glossina morsitans mositans, G.m.submorsitans, G.palpalis gambiensis and G.tachinoides were allowed to feed on rabbits infected with Trypanosoma congolense savannah type or on mice infected with T.congolense riverine-forest type. The four tsetse species and subspecies were also infected simultaneously in vitro on the blood of mice infected with the two clones of T.congolense via a silicone membrane. The infected tsetse were maintained on rabbits and from the day 25 after the infective feed, the surviving tsetse were dissected in order to determine the infection rates. Results showed higher mature infection rates in morsitans-group tsetse flies than in palpalis-group tsetse flies when infected with the savannah type of T.congolense. In contrast, infection rates with the riverine-forest type of T.congolense were lower, and fewer flies showed full development cycle. The intrinsec vectorial capacity of G.m.submorsitans for the two T.congolense types was the highest, whereas the intrinsic vectorial capacity of G.p.gambiensis for the Savannah type and G.m.morsitans for the riverine-forest type were the lowest. Among all tsetse which were infected simultaneously with the two types of T.congolense, the polymerase chain reaction detected only five flies which had both trypanosome taxa in the midgut and the proboscis. All the other infections were attributable to the savannah type. The differences in the gut of different Glossina species and subspecies allowing these two sub-groups of T.congolense to survive better and undergo the complete developmental cycle more readily in some species than other are discussed.