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Background African animal trypanosomosis (AAT), transmitted by tsetse flies, is arguably the main disease constraint to integrated crop-livestock agriculture in sub-Saharan Africa, and African heads of state and governments adopted a resolution to rid the continent of this scourge. In order to sustainably reduce or eliminate the burden of AAT, a progressive and evidence-based approach is needed, which must hinge on harmonized, spatially explicit information on the occurrence of AAT and its vectors. Methods A digital repository was assembled, containing tsetse and AAT data collected in Burkina Faso between 1990 and 2019. Data were collected either in the framework of control activities or for research purposes. Data were systematically verified, harmonized, georeferenced and integrated into a database (PostgreSQL). Entomological data on tsetse were mapped at the level of individual monitoring traps. When this was not possible, mapping was done at the level of site or location. Epidemiological data on AAT were mapped at the level of location or village. Results Entomological data showed the presence of four tsetse species in Burkina Faso. Glossina tachinoides, present from the eastern to the western part of the country, was the most widespread and abundant species (56.35% of the catches). Glossina palpalis gambiensis was the second most abundant species (35.56%), and it was mainly found in the west. Glossina morsitans submorsitans was found at lower densities (6.51%), with a patchy distribution in the southern parts of the country. A single cluster of G. medicorum was detected (less than 0.25%), located in the south-west. Unidentified tsetse flies accounted for 1.33%. For the AAT component, data for 54,948 animal blood samples were assembled from 218 geographic locations. The samples were tested with a variety of diagnostic methods. AAT was found in all surveyed departments, including the tsetse-free areas in the north. Trypanosoma vivax and T. congolense infections were the dominant ones, with a prevalence of 5.19 ± 18.97% and 6.11 ± 21.56%, respectively. Trypanosoma brucei infections were detected at a much lower rate (0.00 ± 0.10%). Conclusions The atlas provides a synoptic view of the available information on tsetse and AAT distribution in Burkina Faso. Data are very scanty for most of the tsetse-free areas in the northern part of the country. Despite this limitation, this study generated a robust tool for targeting future surveillance and control activities. The development of the atlas also strengthened the collaboration between the different institutions involved in tsetse and AAT research and control in Burkina Faso, which will be crucial for future updates and the sustainability of the initiative. Graphical Abstract

Background Tsetse flies are vectors of African trypanosomes, protozoan parasites that cause sleeping sickness (or human African trypanosomosis) in humans and nagana (or animal African trypanosomosis) in livestock. In addition to trypanosomes, four symbiotic bacteria Wigglesworthia glossinidia, Sodalis glossinidius, Wolbachia, Spiroplasma and one pathogen, the salivary gland hypertrophy virus (SGHV), have been reported in different tsetse species. We evaluated the prevalence and coinfection dynamics between Wolbachia, trypanosomes, and SGHV in four tsetse species (Glossina palpalis gambiensis, G. tachinoides, G. morsitans submorsitans, and G. medicorum) that were collected between 2008 and 2015 from 46 geographical locations in West Africa, i.e. Burkina Faso, Mali, Ghana, Guinea, and Senegal. Results The results indicated an overall low prevalence of SGHV and Wolbachia and a high prevalence of trypanosomes in the sampled wild tsetse populations. The prevalence of all three infections varied among tsetse species and sample origin. The highest trypanosome prevalence was found in Glossina tachinoides (61.1%) from Ghana and in Glossina palpalis gambiensis (43.7%) from Senegal. The trypanosome prevalence in the four species from Burkina Faso was lower, i.e. 39.6% in Glossina medicorum, 18.08%; in Glossina morsitans submorsitans, 16.8%; in Glossina tachinoides and 10.5% in Glossina palpalis gambiensis. The trypanosome prevalence in Glossina palpalis gambiensis was lowest in Mali (6.9%) and Guinea (2.2%). The prevalence of SGHV and Wolbachia was very low irrespective of location or tsetse species with an average of 1.7% for SGHV and 1.0% for Wolbachia. In some cases, mixed infections with different trypanosome species were detected. The highest prevalence of coinfection was Trypanosoma vivax and other Trypanosoma species (9.5%) followed by coinfection of T. congolense with other trypanosomes (7.5%). The prevalence of coinfection of T. vivax and T. congolense was (1.0%) and no mixed infection of trypanosomes, SGHV and Wolbachia was detected. Conclusion The results indicated a high rate of trypanosome infection in tsetse wild populations in West African countries but lower infection rate of both Wolbachia and SGHV. Double or triple mixed trypanosome infections were found. In addition, mixed trypanosome and SGHV infections existed however no mixed infections of trypanosome and/or SGHV with Wolbachia were found. Electronic supplementary material The online version of this article (10.1186/s12866-018-1287-4) contains supplementary material, which is available to authorized users.

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

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

Background Tsetse flies are the sole vectors of human and animal trypanosomosis. In Burkina Faso, a project aiming to create zones free of tsetse flies and trypanosomosis was executed from June 2006 to December 2013. After the determination of tsetse distribution in the intervention area from December 2007 to November 2008, the control campaign was launched in November 2009 and ended in December 2013. The goal was to eliminate tsetse flies from 40,000 km2 of area, through an integrated control campaign including insecticide targets, traps and cattle, sequential aerial treatment (SAT) and the mass treatment of livestock using trypanocides. The campaign involved assistance of the beneficiary communities at all the steps of the control strategy with insecticide impregnated targets. Methods This study was carried out to assess the impact of the control project on tsetse apparent density per trap per day (ADT). To evaluate the effectiveness of tsetse control, 201 sites were selected based on the baseline survey results carried out from December 2007 to November 2008. These sites were monitored bi-monthly from January 2010 to November 2012. At the end-of-study in 2013 a generalized entomological survey was carried out in 401 infested sites found during the longitudinal survey done before the control. Barrier and tsetse persistence areas were treated by ground spraying and evaluated. Controls were also done before and after aerial spraying. Results In the insecticide-impregnated target area, the control showed that ADT of tsetse flies declined from 10.73 (SD 13.27) to 0.43 (SD 2.51) fly/trap/day from the third month of campaign onwards (P

In the context of the Pan African Tsetse and Trypanosomiasis Eradication Campaign, the value of tsetse saliva antibodies as a biomarker of cattle exposure to tsetse flies was evaluated, as this could provide an alternative and complementary tool to conventional entomological methods. Serum immune reactivity to Glossina (G.) palpalis (p.)gambiensis, G. tachinoides and G. morsitans (m.) submorsitans whole saliva extracts (WSE) were monitored in cattle from both tsetse free and tsetse infested areas, and in cows experimentally exposed to tsetse flies and other hematophagous arthropods. In the tsetse infested area, cattle IgG responses to Glossina WSE were significantly higher during the dry season (p < 0.0001) when herds are most exposed to tsetse flies and in infected animals (p = 0.01) as expected in the case of a biomarker of exposure. Experimental studies further confirmed this as a quick rise of specific IgGs was observed in animals exposed to tsetse flies (within weeks), followed by a rapid clearance after exposure was stopped. In contrast to the two other tsetse species, G. m. submorsitans WSE enabled to detect exposure to all tsetse species and were associated with low level of cross-reactivity to other blood sucking arthropods. Finally, IgG responses to G. m. submorsitans salivary antigens enabled to distinguish different groups of cows according to exposure levels, thus indicating that tsetse saliva antibodies are not only indicators of tsetse exposure but also are correlated to the intensity of tsetse contacts (p = 0.0031). Implementation of this new sero-epidemiological marker of cattle exposure to tsetse flies in the framework of tsetse elimination campaigns is discussed.

In view of gathering baseline information about the prevalence of animal trypanosomosis, the Pan African Tsetse and Trypanosomiasis Eradication Campaign (PATTEC) funded a cross sectional survey in the region of the Boucle du Mouhoun which constitutes the Northern limit of the tsetse distribution in Burkina. This cross sectional study was carried out in 53 villages located in the six provinces of the region. A total of 2002 cattle, 1466 small ruminants and 481 donkeys were sampled. This survey showed that about 25% of the cattle had been treated with trypanocidal drugs within 3 months before the survey compared to 3% and 0.42% for the small ruminants and donkeys, respectively. Parasitological prevalence in cattle was low: 0.77% (95% C.I. 0.30-1.95%). No goats and three donkeys were found infected with trypanosomes. Infections were mainly due to Trypanosoma vivax (75.0%) with cases of Trypanosoma congolense (25.0%). In cattle, the serological prevalence of trypanosomosis, for the entire region of the Boucle du Mouhoun, was 34.2% (95% C.I. 26.1-43.4%). For sheep, goats and donkeys, the prevalence were of 20.9% (95% C.I. 12.2-33.5%), 8.5% (95% C.I. 5.7-12.5%) and 5.8% (95% C.I. 3.9-8.6%), respectively. The age and distance to the river were the two main risk factors associated with seropositivity.

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

Background The saliva of tsetse flies contains a cocktail of bioactive molecules inducing specific antibody responses in hosts exposed to bites. We have previously shown that an indirect-ELISA test using whole salivary extracts from Glossina morsitans submorsitans was able to discriminate between (i) cattle from tsetse infested and tsetse free areas and (ii) animals experimentally exposed to low or high numbers of tsetse flies. In the present study, our aim was to identify specific salivary synthetic peptides that could be used to develop simple immunoassays to measure cattle exposure to tsetse flies. Methods In a first step, 2D-electrophoresis immunoblotting, using sera from animals exposed to a variety of bloodsucking arthropods, was performed to identify specific salivary proteins recognised in cattle exposed to tsetse bites. Linear epitope prediction software and Blast analysis were then used to design synthetic peptides within the identified salivary proteins. Finally, candidate peptides were tested by indirect-ELISA on serum samples from tsetse infested and tsetse free areas, and from exposure experiments. Results The combined immunoblotting and bioinformatics analyses led to the identification of five peptides carrying putative linear epitopes within two salivary proteins: the tsetse salivary gland protein 1 (Tsal1) and the Salivary Secreted Adenosine (SSA). Of these, two were synthesised and tested further based on the absence of sequence homology with other arthropods or pathogen species. IgG responses to the Tsal152–75 synthetic peptide were shown to be specific of tsetse exposure in both naturally and experimentally exposed hosts. Nevertheless, anti-Tsal152–75 IgG responses were absent in animals exposed to high tsetse biting rates. Conclusions These results suggest that Tsal152–75 specific antibodies represent a biomarker of low cattle exposure to tsetse fly. These results are discussed in the light of the other available tsetse saliva based-immunoassays and in the perspective of developing a simple serological tool for tsetse eradication campaigns to assess the tsetse free status or to detect tsetse reemergence in previously cleared areas. Electronic supplementary material The online version of this article (doi:10.1186/s13071-016-1414-8) contains supplementary material, which is available to authorized users.

The present study was conducted to determine the prevalence and incidence of trypanosomosis and to investigate some factors influencing them in an agro-pastoral area of southwestern Burkina Faso. A total of 363 crossbred cattle (Baoule-zebu peul), which were bred under natural trypanosomosis challenge, were monitored monthly for parasitaemia, packed cell volume (PCV) and serological analyses over 2 years. The parasitological prevalence estimated at the beginning of the survey using the buffy coat technique (BCT) was 7.54%. As much as 66.7% of all trypanosome infections were due to Trypanosoma vivax, 23.8% due to Trypanosoma congolense and 9.5% due to T. vivax/T. congolense mixed infections. The monthly serological incidence varied from 0.29% to 19.29%. The season was the most important factor influencing the serological prevalence and incidence and the animal PCV. The dry hot season is associated with increasing seroprevalences and incidences and consequently a decreasing average of PCV. In addition, an important spatial heterogeneity was observed..

Tsetse-transmitted trypanosomiasis poses a serious threat to human and animal health in sub-Saharan Africa. The majority of tsetse flies (Glossina spp.) in a natural population will not develop a mature infection of either Trypanosoma congolense or Trypanosoma brucei sp. because of refractoriness, a phenomenon that is affected by different factors, including the tsetse fly's immune defence. Starvation of tsetse flies significantly increases their susceptibility to the establishment of a trypanosome infection. This paper reports the effects of nutritional stress (starvation) on (a) uninduced baseline levels of gene expression of the antimicrobial peptides attacin, defensin and cecropin in the tsetse fly, and (b) levels of expression induced in response to bacterial (Escherichia coli) or trypanosomal challenge. In newly emerged, unfed tsetse flies, starvation significantly lowers baseline levels of antimicrobial peptide gene expression, especially for attacin and cecropin. In response to trypanosome challenge, only non-starved older flies showed a significant increase in antimicrobial peptide gene expression within 5 days of ingestion of a trypanosome-containing bloodmeal, especially with T. brucei bloodstream forms. These data suggest that a decreased expression of immune genes in newly hatched flies or a lack of immune responsiveness to trypanosomes in older flies, both occurring as a result of fly starvation, may be among the factors contributing to the increased susceptibility of nutritionally stressed tsetse flies to trypanosome infection.

African animal trypanosomosis (AAT) is considered the most important cattle disease in sub-Saharan Africa but its diagnosis in the field is difficult, resulting in inappropriate treatments, excessive delay in treatments and under-treatment. A field study in West Africa investigated the usefulness of anemia in the diagnosis of trypanosomosis. A total of 20,772 cattle blood samples were taken from 121 villages in 3 countries. The average packed cell volume (PCV) of trypanosomosis positive cattle was 23%, versus 28% for negative cattle. In a sub-set of animals, other causes of anemia were investigated showing most of the anemia burden was attributable to trypanosomosis. Anemia was a reasonably accurate indicator of trypanosomosis in the study area, with a sensitivity of 56% and a specificity of 80% and a diagnostic odds ratio of 4.2, the highest of all the signs evaluated (anemia, emaciation, staring coat, lymphadenopathy, fever, lacrimation and salivary or nasal discharge). Having confirmed the usefulness of anemia as a predictor of trypanosomosis, two potential pen-side tests for anemia were evaluated (the first reported trial of their use in cattle), firstly a color chart developed for anemia detection in sheep through visual inspection of conjunctival membranes (FAMACHA) and secondly the Hemoglobin Color Scale (HbCS) developed for assessing hemoglobin levels in human patients by comparing blood drops on filter paper with color standards. In a population of cattle suspected by their owners to be sick with trypanosomosis (n=898) the sensitivity of the HbCS test was 56% and the specificity was 77%, while the sensitivity of the FAMACHA test was 95% and the specificity was 22%. The higher sensitivity but lower specificity suggests the FAMACHA may be useful as a screening test and the HbCS as a confirmatory test. The two tests were also evaluated in cattle randomly selected from the village herd. Using cut-off points to optimize test performance, the HbCS test had a sensitivity of 81% and a specificity of 62% (n=505 cattle), while the FAMACHA had a sensitivity of 92% and a specificity of 30% (n=298 cattle). Recommendations are made for the appropriate use of these tests in the West African region.

Background The application of the sterile insect technique (SIT) requires mass-production of sterile males of good biological quality. The size of the project area will in most cases determine whether it is more cost effective to produce the sterile flies locally (and invest in a mass-rearing facility) or import the sterile flies from a mass-rearing facility that is located in another country. This study aimed at assessing the effect of long distance transport of sterile male Glossina palpalis gambiensis pupae on adult male fly yield. Methods The male pupae were produced at the Centre International de Recherche-Développement sur l’Elevage en zone Subhumide (CIRDES), Bobo-Dioulasso, Burkina Faso, and shipped with a commercial courier service in insulated transport boxes at a temperature of ±10°C to Senegal (±36 h of transport). Upon arrival in the insectary in Dakar, the pupae were transferred to an emergence room and the flies monitored for 3–6 days. Results The results showed that the used system of isothermal boxes that contained phase change material packs (S8) managed to keep the temperature at around 10°C which prevented male fly emergence during transport. The emergence rate was significantly higher for pupae from batch 2 (chilled at 4°C for one day in the source insectary before transport) than those from batch 1 (chilled at 4°C for two days in the source insectary before transport) i.e. an average (±sd) of 76.1 ± 13.2% and 72.2 ± 14.3%, respectively with a small proportion emerging during transport (0.7 ± 1.7% and 0.9 ± 2.9%, respectively). Among the emerged flies, the percentage with deformed (not fully expanded) wings was significantly higher for flies from batch 1 (12.0 ± 6.3%) than from batch 2 (10.7 ± 7.5%). The amount of sterile males available for release as a proportion of the total pupae shipped was 65.8 ± 13.3% and 61.7 ± 14.7% for batch 1 and 2 pupae, respectively. Conclusions The results also showed that the temperature inside the parcel must be controlled around 10°C with a maximal deviation of 3°C to maximize the male yield. Electronic supplementary material The online version of this article (doi:10.1186/s13071-015-0869-3) contains supplementary material, which is available to authorized users.

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

Resistance to the drugs used to control African animal trypanosomosis is increasingly recognised as a constraint to livestock production in sub-Saharan Africa. The most commonly used tests for detection of trypanocidal drug resistance are tests using mice or ruminants, but these suffer from lack of standardisation and hence it may be difficult to compare the results of different investigators. Tests in mice are less expensive than tests in ruminants, but while tests in mice they may be useful as a general guide to resistance in a geographic area they should not be extrapolated to cattle on an individual trypanosome level. Moreover, the commonly used protocols are too laborious for their application to large number of trypanosome isolates on an area-wide basis. This paper presents guidelines for standardised testing of trypanocidal drugs in vivo, and introduces a simplified single-dose test for use in mice, which is convenient for use in areas with limited laboratory facilities. The single-dose test is appropriate for characterisation of geographic areas in terms of trypanocidal drug resistance using large numbers of trypanosome isolates, for making comparisons between areas, and for monitoring changes in trypanocidal drug resistance over time. Multiple-dose tests may be used to determine the degree of resistance of individual stabilates to be determined precisely in mice are also described, but for logistical reasons these will rarely be conducted on more than a few stabilates, and testing of a larger number of stabilates in the single-dose test will generally provide more useful information. Finally, we describe tests in cattle that may be used to determine the efficacy of recommended curative doses of trypanocidal drugs for the treatment of infection with individual trypanosome isolates, including Trypanosoma vivax, which is rarely infective for mice.

Blood was collected from two Sahelian goats, experimentally infected with either a drug-sensitive cloned population of Trypanosoma congolense (IL 1180) or a multiple drug-resistant T. congolense stock (Samorogouan/89/CRTA/267) and incubated at 37 degrees C for 30 min and 12 h, respectively, in the presence of different drug concentrations (0.5, 1.0, 10.0 and 100.0 microg/ml blood) of diminazene aceturate or isometamidium chloride. After that, the trypanosome/blood/drug suspensions were offered to tsetse flies (2100 teneral Glossina morsitans submorsitans) through an in vitro feeding system, using a silicone membrane. All tsetse flies were dissected and examined for the presence of trypanosomes in labrum, hypopharynx and midgut 20 days after their infective blood-meals. Infectivity of the drug-sensitive cloned population was already completely abolished after incubation with 0.5 microg/ml of both drugs; however, 13.6-42.2% of tsetse having been fed on untreated blood had developed an infection. In contrast, no significant differences were observed in the infection rates between the experimental groups and their control groups when fed on blood infected with the multiple drug-resistant stock after incubation for 30 min with up to 10 microg/ml of diminazene or isometamidium. In consequence, tsetse appear to be a useful tool in the assessment of drug susceptibility of typanosome populations.

Background Tsetse flies are the cyclical vectors of African trypanosomosis that constitute a major constraint to development in Africa. Their control is an important component of the integrated management of these diseases, and among the techniques available, the sterile insect technique (SIT) is the sole that is efficient at low densities. The government of Burkina Faso has embarked on a tsetse eradication programme in the framework of the PATTEC, where SIT is an important component. The project plans to use flies from a Glossina palpalis gambiensis colony that has been maintained for about 40 years at the Centre International de Recherche-Developpement sur l'Elevage en zone Subhumide (CIRDES). It was thus necessary to test the competitiveness of the sterile males originating from this colony. Methodology/Principal Findings During the period January-February 2010, 16,000 sterile male G. p. gambiensis were released along a tributary of the Mouhoun river. The study revealed that with a mean sterile to wild male ratio of 1.16 (s.d. 0.38), the abortion rate of the wild female flies was significantly higher than before (p = 0.026) and after (p = 0.019) the release period. The estimated competitiveness of the sterile males (Fried index) was 0.07 (s.d. 0.02), indicating that a sterile to wild male ratio of 14.4 would be necessary to obtain nearly complete induced sterility in the female population. The aggregation patterns of sterile and wild male flies were similar. The survival rate of the released sterile male flies was similar to that observed in 1983-1985 for the same colony. Conclusions/Significance We conclude that gamma sterilised male G. p. gambiensis derived from the CIRDES colony have a competitiveness that is comparable to their competitiveness obtained 35 years ago and can still be used for an area-wide integrated pest management campaign with a sterile insect component in Burkina Faso. (Resume d'auteur)

Trypanocidal drug resistance is unanimously recognized as a threat for livestock production in regions where the prevalence of trypanosomosis is high. To assess the impact of the disease and the effect of drug resistance on the health of small ruminants, twelve Trypanosoma vivax isolates collected in 6 villages in the vicinity of Bobo Dioulasso (Burkina Faso) were injected into 12 groups of 5 Sahelian goats, two being treated with 3.5 mg/kg body weight diminazene aceturate (DA), two with 0.5 mg/kg body weight isometamidium chloride (ISM) and one left untreated as control. A monitoring was performed every 5 days for 100 days to evaluate the parasitaemia by buffy coat examination, the hematocrit and the body weight. Among the 12 groups, 6 were additionally monitored using a trypanosome specific 18S-PCR–RFLP every 5 days from day 30 to day 100 to verify the complete clearance of the parasites from the blood of the hosts. In six groups of goats, trypanosomes disappeared completely after treatment, five groups showed relapses in at least one goat treated with ISM and one group showed relapses in one goat treated with DA and one with ISM. For the 6 groups that were screened both using microscopic examination and trypanosome specific 18S-PCR–RFLP, the following results were observed: for the groups treated with DA, no relapses by microscopic examination and 83.3% (10/12) using the 18S-PCR–RFLP. For the groups treated with ISM, 25% (3/12) relapses by microscopic examination and 83.3% with the 18S-PCR–RFLP (10/12). The evolution of the PCV and the weight during the observation period from relapsing (either by microscopical examination or by 18S-PCR–RFLP diagnosis) and non relapsing animals were compared. The relative average PCV in goats that relapsed microscopically, decreased significantly more than in non-relapsing goats. This difference was not significant when relapses were detected using the trypanosome specific 18S-PCR–RFLP. This indicates that only the animals with the highest parasitaemia suffered from the infection. Relapses after treatment where the host controls the parasitaemia to a level below the sensitivity of the microscopical examination do not affect body weight nor PCV.

A longitudinal study assessed the chemoresistance to isometamidium chloride (ISM) and diminazene aceturate (DA) in the region of the Boucle du Mouhoun in Burkina Faso. A preliminary cross-sectional survey allowed the identification of the 10 villages with the highest parasitological prevalences (from 2.1% to 16.1%). In each of these 10 villages, two herds of approximately 50 bovines were selected, one being treated with ISM (1mg/kg b.w.) and the other remaining untreated as control group. All animals (treated and untreated herds) becoming infected were treated with DA (3.5mg/kg b.w.). In total, 978 head of cattle were followed up. Fortnightly controls of the parasitaemia and PCV were carried out during 8 weeks. The main trypanosome species was Trypanosoma vivax (83.6%) followed by Trypanosoma congolense (16.4%). In two villages, less than 25% of the control untreated cattle became positive indicating no need to use prophylactic treatment. These two villages were not further studied. Resistance to ISM was observed in 5 of the remaining 8 villages (Debe, Bendougou, Kangotenga, Mou and Laro) where the relative risk (control/treated hazard ratios) of becoming infected was lower than 2 i.e. between 0.89 (95% CI: 0.43-2.74) and 1.75 (95% CI: 0.57-5.37). In contrast, this study did not show evidence of resistance to DA in the surveyed villages with only 8.6% (n=93) of the cattle relapsing after treatment. Our results suggest that because of the low prevalence of multiple resistances in the area a meticulous use of the sanative pair system would constitute the best option to delay as much as possible the spread of chemoresistance till complete eradication of the disease by vector control operations.

International audience; 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.

Animal African trypanosomiasis (AAT) is caused by different species of the protozoan parasite Trypanosoma and affects a wide range of domestic animals. Trypanosoma congolense is widespread in the whole of sub-Saharan Africa and is the species causing considerable losses in livestock production, often affecting the health status of humans through endangering the food supply of rural communities. It is estimated that 50 million cattle are at risk of the disease and that the direct and indirect annual losses related to AAT reach US$4.5 billion [1]. Despite ambitious AAT control programmes such as the Pan African Tsetse and Trypanosomiasis Eradication Campaign that aim to eradicate the disease from the African continent, the control of the disease will in the foreseeable future continue to rely heavily on the use of trypanocidal drugs. Diminazene aceturate (DA) is the cheapest and most popular trypanocide. However, this drug, commercialized more than half a century ago, tends to become ineffective due to the spread of trypanocidal drug resistance now reported in 17 African countries [2]. The different techniques for the diagnosis of trypanocidal drug resistance were exhaustively reviewed by Delespaux et al. [2]. Two techniques are routinely used in the field namely, block treatments [3] and in vivo tests [4]. Block treatment involves follow-up for a period of 2 months of treated and non-treated (control) herds. This method provides information on the hazard of contracting trypanosomiasis (number of cattle becoming positive in the control group) and on the level of trypanocidal drug resistance (number of cattle becoming positive in the treated group). In vivo tests (performed in small ruminants for Trypanosoma vivax or in rodents for T. congolense) remain the gold standards but present some drawbacks, such as (i) problems associated with the growth of some T. congolense strains in rodents, (ii) the long period of follow up (2 months), and (iii) the intensive use of experimental animals. To address these important drawbacks, the Department of Animal Health of the Institute of Tropical Medicine Antwerp has been actively involved in research on trypanocidal drug resistance and in 2007 was designated Reference Centre for Livestock Trypanosomiasis, Parasite Management and Diagnosis (RCLT) by the Food and Agriculture Organization of the United Nations (FAO) and accredited to ISONORM 17025 in 2009. Here, a BclI-PCR-RFLP was developed for the molecular diagnosis of DA resistance in T. congolense [5], the most widely used trypanocidal drug. The test is based on the detection of a single point mutation in P2-type purine transporters and has been transferred to a network of regional reference laboratories in West, East, and Southern Africa for use in large-scale surveys to detect the presence of trypanocidal drug resistance throughout tsetse-infested Africa. The correlation of this PCR-RFLP with the single dose mouse test (gold standard) [4] is excellent even if the PCR-RFLP was found to be more sensitive than the single dose mouse test [5], [6]. Although the test performs very well under laboratory conditions [6], it requires further adaptation and evaluation for use under field conditions. More specifically, the test's ability to amplify low concentrations of parasite DNA (as a result of often low parasitaemias in livestock) needs to be enhanced and its specificity improved by preventing incomplete digestion of the amplicon showing the point mutation by the BclI enzyme. This incomplete digestion creates profiles consisting of partially undigested and digested amplicons (resistant) and as such, falsely mixed RFLP profiles. Finally, sample collection, storage, and transfer to regional reference laboratories need to be simplified. In view of the above, the test's ability to amplify low concentrations of DNA was enhanced by adding a step of whole genome amplification using the QIAGEN REPLI-g UltraFast Mini Kit. Its specificity was improved by replacing the BclI (T∧GATCA) enzyme by DpnII (∧GATC). Internal negative and positive controls (resistant and sensitive strains characterized by sequencing their respective amplicons) were added to ensure the absence of contamination, an effective DNA amplification, and the complete digestion of the PCR products. Finally, sample collection, storage, and transfer could be facilitated by collecting blood spots or buffy coats on filter paper (Whatman N°4, Whatman) (Figures 1 and ​and2).2). When costs and availability are not a limitation, the use of Whatman FTA or FTA Elute cards could constitute an interesting alternative, as it is believed to offer superior preservation of the DNA. Figure 1 Laboratory material used for the processing of the samples. Figure 2 Dried blood spots (left) and buffy coats (right) on labelled filter papers. To evaluate the performance of the improved PCR-RFLP using the DpnII enzyme for the detection of DA resistance in T. congolense, use was made of 449 whole blood spots (40–50 µl) on filter papers collected from parasitologically positive cattle originating from the cotton zone of Southern Mali where significant resistance to trypanocidal drugs was reported [7]. The samples were stored sun-protected for 1 year at tropical ambient temperature without adding dehydrated silica crystals in the storage plastic bags (i.e., stored under suboptimal conditions as often is the case in rural environments), allowing degradation of the DNA. A total of 68% (304) of all blood spots was found positive for the presence of trypanosomes using the 18S pan-PCR developed by Geysen et al. [8]. Of these, 74% (225) were diagnosed as a single or mixed T. congolense infection. Out of these 225 T. congolense–positive samples, 26% (59) amplified using the DPnII-PCR-RFLP. By adding a step of whole genome amplification, an extra 42 samples became positive, reaching a total of 44.9% (101). The DPnII-PCR-RFLP profiles of the 101 amplified T. congolense samples were distributed as follows: 92% were resistant, 2% sensitive, and 6% presented a mixed profile (Figure 3). Even if due to host-related factors not all animals with those infections would fail to respond to treatment, such a high prevalence of DA resistance is more than worrying. As proposed by Clausen et al. [7], in such circumstances an integrated approach should be used based on (i) rational drug use information given to farmers, (ii) training farmers and paravets in integrated trypanosomiasis control, and (iii) training animal care providers to a more specific diagnosis of trypanosomiasis to avoid treatments inappropriate for the clinical signs. Figure 3 Output of the DPnII-PCR-RFLP with lanes 1, 6, and 10 as size markers (100-bp ladder), lanes 2, 3, 4, 5, and 7 as sensitive profiles (one band), lane 8 as resistant profile (two bands), and lane 9 as mixed profile (three bands). Notwithstanding the suboptimal storage procedure, it can be concluded that blood samples collected and stored on filter papers can be used for detecting the presence of trypanosomes resistant to DA in a trypanosome population. Even under suboptimal conditions, 44.9% of the T. congolense samples were successfully characterized for DA resistance sufficient to estimate the prevalence and repartition of DA resistance in the field. Moreover, losses due to suboptimal storage conditions can be compensated for by increasing the sample size. Hence, considering the important logistical and economic advantages of filter papers, their use greatly facilitates the implementation of large-scale surveys for trypanocidal drug resistance in trypanosomiasis-affected African countries using molecular diagnostic tools. Moreover, their use greatly supports the functioning and sustainability of much-needed regional reference laboratories. Box 1. Advantages and Disadvantages of using the DpnII-PCR-RFLP for the Diagnosis of DA Resistance in T. congolense on Blood Samples Collected on Filter Papers Advantages Easy collection/storage/transfer of samples (blood spots or buffy coats on filter paper stored at ambient room temperature). Allows even for the screening of large numbers of trypanosome strains circulating in remote areas without use of experimental animals. Contributes to the sustainability of regional reference laboratories for trypanocidal drug resistance. Disadvantages Incorrect storage of filter papers reduces the number of PCR-positive samples. The use of filter papers increases the danger of sample contamination. Relatively high cost (25€ for species identification and DA resistance detection, all included except technician salary and transport) but still acceptable compared to a test in mice (150€ all included except technician salary and transport), tests in cattle or block treatment being even more expensive (e.g., €945 for a survey of treatment failure in a village herd).

The northern distribution limit of tsetse flies was updated in Burkina Faso and compared to previous limits to revise the existing map of these vectors of African trypanosomiases dating from several decades ago. From 1949 to 2009, a 25- to 150-km shift has appeared toward the south. Tsetse are now discontinuously distributed in Burkina Faso with a western and an eastern tsetse belt. This range shift can be explained by a combination of decreased rainfall and increased human density. Within a context of international control, this study provides a better understanding of the factors influencing the distribution of tsetse flies.

Preliminary data from an ongoing epidemiological survey in the pastoral zone of Samorogouan (Kénédougou) indicate the occurrence of multiple-drug-resistant Trypanosoma congolense. Despite frequent trypanocidal drug treatments with diminazene aceturate (Berenil, Hoechst) at 7 mg/kg body weight (bw) at intervals of 2 to 4 weeks, no significant drop in the prevalence of African animal trypanosomosis (AAT) was observed. To examine a suspected drug resistance, 20 Zebu cattle, naturally infected with T. congolense and/or T. vivax, were transferred in December 1989 from Samorogouan into a fly-proof stable. Diminazene aceturate at 7 mg/kg bw cured infections of T. vivax, but was ineffective against T. congolense. Likewise, treatments with homidium bromide (Ethidium, FBC) at 1 mg/kg bw and isometamidium chloride (Trypamidium, Rhône Mérieux) at 1 mg/kg bw, respectively, proved to be ineffective. Corresponding chemotherapeutic trials in previously unexposed Zebu bulls and Sahelian goats infected with one primary T. congolense isolate from Samorogouan demonstrated a high level of resistance to diminazene aceturate (7 mg/kg bw in cattle and 17.5 mg/kg bw in goats), isometamidium chloride (1 and 2 mg/kg bw i.v. in goats) and quinapyramine sulphate (Trypacide'S', Rhône Mérieux) at 5 mg/kg bw in goats. The appearance of a multiple-drug-resistant strain of T. congolense emphasizes the urgent need for new chemical substances as trypanocidal drugs and the increasing importance of efficient vector control.

African animal trypanosomosis is a major obstacle to the development of more efficient and sustainable livestock production systems in West Africa. Riverine tsetse species such as Glossina palpalis gambiensis Vanderplank are their major vectors. A wide variety of control tactics is available to manage these vectors, but their elimination will only be sustainable if control is exercised following area-wide integrated pest management (AW-IPM) principles, i.e. the control effort is targeting an entire tsetse population within a circumscribed area. In the present study, genetic variation at microsatellite DNA loci was used to examine the population structure of G. p. gambiensis inhabiting two adjacent river basins, i.e. the Comoe and the Mouhoun River basins in Burkina Faso. A remote sensing analysis revealed that the woodland savannah habitats between the river basins have remained unchanged during the last two decades. In addition, genetic variation was studied in two populations that were separated by a man-made lake originating from a dam built in 1991 on the Comoe. Low genetic differentiation was observed between the samples from the Mouhoun and the Comoe River basins and no differentiation was found between the samples separated by the dam. The data presented indicate that the overall genetic differentiation of G. p. gambiensis populations inhabiting two adjacent river basins in Burkina Faso is low (FST = 0.016). The results of this study suggest that either G. p. gambiensis populations from the Mouhoun are not isolated from those of the Comoe, or that the isolation is too recent to be detected. If elimination of the G. p. gambiensis population from the Mouhoun River basin is the selected control strategy, re-invasion from adjacent river basins may need to be prevented by establishing a buffer zone between the Mouhoun and the other river basin(s).

International audience; The West African trypanosomoses are mostly transmitted by riverine species of tsetse fly. In this study, we estimate the dispersal and population size of tsetse populations located along the Mouhoun river in Burkina Faso where tsetse habitats are experiencing increasing fragmentation caused by human encroachment. Dispersal estimated through direct (mark and recapture) and indirect (genetic isolation by distance) methods appeared consistent with one another. In these fragmented landscapes, tsetse flies displayed localized, small subpopulations with relatively short effective dispersal. We discuss how such information is crucial for designing optimal strategies for eliminating this threat. To estimate ecological parameters of wild animal populations, the genetic measures are both a cost- and time-effective alternative to mark-release-recapture. They can be applied to other vector-borne diseases of medical and/or economic importance.

The impact of landscape fragmentation due to human and climatic mediated factors on the structure of a population of Glossina palpalis gambiensis Vanderplank (Diptera: Glossinidae) was investigated in the Mouhoun river basin, Burkina Faso. Allele frequencies at five microsatellite loci, and metric properties based on 11 wing landmarks, were compared between four populations. The populations originated from the Mouhoun river and one of its tributaries. The average distance between samples was 72 km with the two most widely spaced populations being 216 km apart. The sampling points traversed an ecological cline in terms of rainfall and riverine forest ecotype, along a river enlarging from downstream to upstream and oriented south to north. Microsatellite DNA comparison demonstrated structuring between the populations, but not complete isolation, with an overall Fst = 0.012 (P0.001). Wing geometry revealed significant centroid size and shape differences between populations, especially between the two most distant populations. There was no significant correlation between gene flow and geographic distance at this scale, but there was a positive correlation in females between metric distances (wing shape differences) and geographic distances that might be attributed to the cline of environmental conditions. The impact of the fragmentation of riparian landscapes on tsetse population structure is discussed in the context of control campaigns currently promoted by Pan African Tsetse and Trypanosomosis Eradication Campaign.

Summary Several previous studies concluded that linkage disequilibrium (LD) in livestock populationsfrom developed countries originated from the impact of strong selection. Here, we assessedthe extent of LD in a cattle population from western Africa that was bred in an extensivefarming system. The analyses were performed on 363 individuals in a Bos indicus · Bostaurus population using 42 microsatellite markers on BTA04, BTA07 and BTA13. A highlevel of expected heterozygosity (0.71), a high mean number of alleles per locus (9.7) and amild shift in Hardy–Weinberg equilibrium were found. Linkage disequilibrium extendedover shorter distances than what has been observed in cattle from developed countries.Effective population size was assessed using two methods; both methods produced largevalues: 1388 when considering heterozygosity (assuming a mutation rate of 10 )3 ) and2344 when considering LD on whole linkage groups (assuming a constant population sizeover generations). However, analysing the decay of LD as a function of marker spacingindicated a decreasing trend in effective population size over generations. This decreasecould be explained by increasing selective pressure and/or by an admixture process. Finally,LD extended over small distances, which suggested that whole-genome scans will require alarge number of markers. However, association studies using such populations will beeffective.Keywords cattle, effective population size, linkage disequilibrium, microsatellites, popu-lation history.

The aim of this study was to assess, whether polymerase chain reaction (PCR) allows sensitive screening of treatment failure suspicions in areas, where drug resistance against African animal trypanosomosis (AAT) appears to be a problem. PCR was used to detect trypanosome infections prior to, 14 and 28 days after controlled treatment of 738 cattle from 10 villages in Kenedougou, Burkina Faso with isometamidium chloride and diminazene aceturate. Using three sets of primers, PCR was three-four times more sensitive and better at species identification, than standard microscopic examination. The better sensitivity and species specificity of PCR have important advantages for drug resistance studies in the field.

Field studies were conducted to assess the occurrence of resistance to isometamidium chloride and diminazene aceturate in trypanosomes infecting cattle in Kenedougou Province of Burkina Faso. Forty-five of the 166 villages in Kenedougou were randomly sampled and visited to assess livestock numbers, trypanosomosis risk, and tsetse challenge. The proportion of cattle infections associated with drug-resistant trypanosomes was assessed in the nine villages with the highest trypanosome infection prevalence and one village with a confirmed history of drug-resistant infections. These studies showed that resistance to both isometamidium and diminazene was widespread. However, there was considerable variation between villages in drug-resistance parameters, with the proportion of treated cattle with trypanosome infections 3 months after isometamidium prophylaxis varying from 6.9 to 63.8% and the proportion of cattle having infections 2 weeks after treatment with diminazene varying from 0 to 36.8%. The demonstration of widespread resistance to both isometamidium and diminazene has important implications, as administration of trypanocides is the most commonly employed method to control trypanosomosis in this area.

Bovine dermatophilosis is a severe skin infection of tropical ruminants inducing a severe loss in productivity and a 15% mortality rate. This disease is caused by the actinomycete bacterium Dermatophilus congolensis associated with the tick Amblyomma variegatum. Currently there are no prospects for a vaccine, and acaricide or antibiotic control is hampered by the development of chemoresistance. Animal breeders have observed that dermatophilosis susceptibility seems to be determined genetically, and we previously identified a BoLA-DRB3-DQB class II haplotype marker for high (R2 = 0.96) susceptibility to the disease. With this marker, we developed a successful eugenic selection procedure for zebu Brahman cattle in Martinique (FWI). Over a period of five years, a marked reduction in disease prevalence, from 0.76 to 0.02 was achieved, and this low level has been maintained over the last two years. The selection procedure, based on a genetic marker system targeting the highly polymorphic BoLA locus, eliminates only those individuals which are at the highest risk of contracting the disease. In the present work, we discuss the properties of this system, including the "heterozygote advantage" and the "frequency dependence" theories, and examine their involvement in the biological mechanisms at the host/pathogen interface. We speculate on the exact role of the MHC molecules in the control of the disease, how the natural selection pressure imposed by the pathogens selectively maintains MHC diversity, and how our results can be practically applied for integrated control of dermatophilosis in developing countries.

Inbred Balb/c mice were infected with three clones of Trypanosoma congolense (Sam.28.1, Dind.3.1 and K60.1A) corresponding, respectively, to the three genetically distinct types (savannah, forest and kilifi) defined within this species, for the purpose of comparing their pathogenicity for a better understanding of the epidemiology of African trypanosomosis. Another clone of savannah type, IL 3000, was also tested simultaneously to study a probable strain variation. Both the clones of savannah type were found of extreme virulence with loss of appetite, rough hair, rapid respiration, lethargy, and all mice died within a week. Parasitaemias evolved rapidly to the first peak by day 3-5 post-inoculation without any remission and the course of disease was correlated positively with the prepatent period. The clones of the forest type and the kilifi type were of low virulence with chronic infection and symptoms progressively less patent throughout the infection; only one mouse died in each experimental group.

The pathology of African bovine trypanosomosis was compared in Zebu cattle subcutaneously inoculated with three clones of trypanosomes corresponding to the three genetically distinct types of Trypanosoma congolense; savannah-type, west African riverine/forest-type and kilifi-type. All inoculated animals became parasitaemic between 7 and 11 days post-infection (dpi). The savannah-type showed consistently higher levels of parasitaemia and lower packed red cell volume percentages and leukocyte counts than the other two types. The syndrome was also more severe in the savannah-type and led inexorably to death between 29 and 54 dpi while animals with the forest or the kilifi-types recovered from earlier symptoms and haematological alterations after 3 months of infection. By the end of the experiment, the animals self-cured from the forest-type infection and the kilifi-type passed under control. The results of the present study indicated clear difference in pathogenicity between the three types of T. congolense; the savannah-type was virulent while the forest-type was of low pathogenicity and the kilifi-type was non-pathogenic.

The diagnostic performance of a polymerase chain reaction assay (PCR) for monitoring the effectiveness of aceturate diminazene treatment was compared with those of an antibody-detection ELISA test and the buffy-coat technique using sheep experimentally infected with either savannahtype or forest-type Trypanosoma congolense or T. vivax. Within the period of infection, the PCR using specific savannah-type T. congolense primers showed a significant higher diagnostic sensitivity (p < 0.05) than the buffy-coat technique. Both techniques gave closed results for detecting forest-type T. congolense or T. vivax infections. Following trypanocidal treatment, the PCR showed that specific product disappeared definitively 1 or 2 days later in animals in which a decrease of the antibody level and a significant improvement of the red packed cell volume were observed. The occurrence of relapse infection was detected by the PCR in one animal infected by T. vivax on day 19 post-treatment and confirmed by the persistence and increasing antibody level whereas the buffy-coat technique detected parasites 42 days later. Then, the PCR signals remained positive on several occasions while parasitaemia was detected only two times. The application of PCR combined with the antibody detection appeared to provide a useful tool as compared to the buffy-coat technique for monitoring the effectiveness of trypanocidal treatment.

The epidemiology of animal trypanosomosis around Bobo-Dioulasso (Burkina Faso, West Africa) benefitted 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 the 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 information 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. (Résumé d'auteur)

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.

We have assessed the phylogenetic status of the #Trypanosoma cruzi$ Genome Project CL Brener reference strain by multilocus enzyme electrophoresis (MLEE) and multiprimer random amplified polymorphic DNA (RAPD) including a set of cloned stocks representative of the whole genetic diversity of #T. cruzi$. MLEE and RAPD data gave congruent phylogenetic results. The CL Brener reference strain fell into the second major phylogenetic subdivision of #T. cruzi$, and was genetically very close to the Tulahuen reference strain. No reliable RAPD character and only one MLEE character permitted us to distinguish between the CL Brener and Tulahuen reference strain. In contrast, many RAPD and MLEE characters were able to distinguish between the CL Brener reference strain and the other #T. cruzi$ genotypes analyzed here, in particular the formerly described principal zymodemes I, II and III. It is suspected that both CL Brener and Tulahuen are hybrid genotypes, a fact that should be taken into account when interpreting sequence data. Moreover, our study confirms that the species #T. cruzi$ is genetically very heterogeneous. We recommend future comparison of sequencing data from the CL Brener reference strain with those of at least one radically distinct #T. cruzi$ genotype, belonging to the other major phylogenetic subdivision of this species. (Résumé d'auteur)