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Auty H.K.,University of Edinburgh | Auty H.K.,University of Glasgow | Picozzi K.,University of Edinburgh | Malele I.,Tsetse and Trypanosomiasis Research Institute | And 3 more authors.
PLoS Neglected Tropical Diseases | Year: 2012

Background: Measuring the prevalence of transmissible Trypanosoma brucei rhodesiense in tsetse populations is essential for understanding transmission dynamics, assessing human disease risk and monitoring spatio-temporal trends and the impact of control interventions. Although an important epidemiological variable, identifying flies which carry transmissible infections is difficult, with challenges including low prevalence, presence of other trypanosome species in the same fly, and concurrent detection of immature non-transmissible infections. Diagnostic tests to measure the prevalence of T. b. rhodesiense in tsetse are applied and interpreted inconsistently, and discrepancies between studies suggest this value is not consistently estimated even to within an order of magnitude. Methodology/Principal Findings: Three approaches were used to estimate the prevalence of transmissible Trypanosoma brucei s.l. and T. b. rhodesiense in Glossina swynnertoni and G. pallidipes in Serengeti National Park, Tanzania: (i) dissection/microscopy; (ii) PCR on infected tsetse midguts; and (iii) inference from a mathematical model. Using dissection/microscopy the prevalence of transmissible T. brucei s.l. was 0% (95% CI 0-0.085) for G. swynnertoni and 0% (0-0.18) G. pallidipes; using PCR the prevalence of transmissible T. b. rhodesiense was 0.010% (0-0.054) and 0.0089% (0-0.059) respectively, and by model inference 0.0064% and 0.00085% respectively. Conclusions/Significance: The zero prevalence result by dissection/microscopy (likely really greater than zero given the results of other approaches) is not unusual by this technique, often ascribed to poor sensitivity. The application of additional techniques confirmed the very low prevalence of T. brucei suggesting the zero prevalence result was attributable to insufficient sample size (despite examination of 6000 tsetse). Given the prohibitively high sample sizes required to obtain meaningful results by dissection/microscopy, PCR-based approaches offer the current best option for assessing trypanosome prevalence in tsetse but inconsistencies in relating PCR results to transmissibility highlight the need for a consensus approach to generate meaningful and comparable data. © 2012 Auty et al. Source

Malele I.I.,Tsetse and Trypanosomiasis Research Institute
Tanzania Journal of Health Research | Year: 2011

Tsetse flies are the vectors of trypanosomes, the causative organisms of trypanosomiasis, nagana, in animals and sleeping sickness in man. In Tanzania, tsetse transmitted trypanosomiasis is one of the most important disease affecting both animals and humans. About 40% of land suitable for grazing and areas with high agricultural potential are currently tsetse infested. It is estimated that about 4.4 million livestock and 4 million people are at risk of contracting tsetse borne trypanosomiasis. African animal trypanosomiasis (AAT) causes loss in animals due to mortality and reduced milk yield, which is estimated at US$ 7.98 million annually. Even after 50 years of independence, Human African Trypanosomiasis (HAT) or Sleeping Sickness is still one of the major public health problems with about 300 cases being reported annually. Tsetse control has been sporadic and uncoordinated hence no tangible results have been accrued since independence despite the fact that technologies which have facilitated tsetse control in other places are available. Fifty years of independence have seen shrinkage of tsetse belt to 43% in 16 surveyed regions. Opportunities for future are wide open if tsetse control will involve all stakeholders, who are directly or indirectly affected by the tsetse problem; if tsetse and trypanosomiasis eradication will adopt an area wide and participatory approach with emphasis on environmentally and user friendly techniques for expanded livestock sector; improved food security and livelihood in affected communities, for achievement of the millennium development goals. Source

Enyaru J.C.,Makerere University | Ouma J.O.,Kenya Agricultural Research Institute | Malele I.I.,Tsetse and Trypanosomiasis Research Institute
Trends in Parasitology | Year: 2010

Understanding what the trypanosome pathogens are, their vectors and mode of transmission underpin efforts to control the disease they cause in both humans and livestock. The risk of transmission is estimated by determining what proportion of the vector population is carrying the infectious pathogens. This risk also depends on the infectivity of the trypanosomes to humans and livestock. Most livestock pathogens are not infective to humans, whereas the two sub-species that infect humans also infect livestock. As with other infectious diseases, we can therefore trace the foundation of many continuing disease control programs for trypanosomiasis to the discovery of the pathogens and their vectors more than a century ago. Over this period, methods for detecting and identifying trypanosomes have evolved through various landmark discoveries. This review describes the evolution of methods for identifying African trypanosomes in their tsetse fly vectors. © 2010 Elsevier Ltd. Source

Ouma J.O.,Kenya Agricultural Research Institute | Malele I.I.,Tsetse and Trypanosomiasis Research Institute | Ngure R.M.,Egerton University | Rutto J.J.,Kenya Agricultural Research Institute | Enyaru J.,Makerere University
PLoS ONE | Year: 2011

Tsetse flies are notoriously difficult to observe in nature, particularly when populations densities are low. It is therefore difficult to observe them on their hosts in nature; hence their vertebrate species can very often only be determined indirectly by analysis of their gut contents. This knowledge is a critical component of the information on which control tactics can be developed. The objective of this study was to determine the sources of tsetse bloodmeals, hence investigate their feeding preferences. We used mitochondrial cytochrome c oxidase 1 (COI) and cytochrome b (cytb) gene sequences for identification of tsetse fly blood meals, in order to provide a foundation for rational decisions to guide control of trypanosomiasis, and their vectors. Glossina swynnertoni were sampled from Serengeti (Tanzania) and G. pallidipes from Kenya (Nguruman and Busia), and Uganda. Sequences were used to query public databases, and the percentage identities obtained used to identify hosts. An initial assay showed that the feeds were from single sources. Hosts identified from blood fed flies collected in Serengeti ecosystem, included buffaloes (25/40), giraffes (8/40), warthogs (3/40), elephants (3/40) and one spotted hyena. In Nguruman, where G. pallidipes flies were analyzed, the feeds were from elephants (6/13) and warthogs (5/13), while buffaloes and baboons accounted for one bloodmeal each. Only cattle blood was detected in flies caught in Busia and Uganda. Out of four flies tested in Mbita Point, Suba District in western Kenya, one had fed on cattle, the other three on the Nile monitor lizard. These results demonstrate that cattle will form an integral part of a control strategy for trypanosomiasis in Busia and Uganda, while different approaches are required for Serengeti and Nguruman ecosystems, where wildlife abound and are the major component of the tsetse fly food source. © 2011 Muturi et al. Source

Nakao R.,Hokkaido University | Stromdahl E.Y.,U.S. Army | Magona J.W.,Hokkaido University | Magona J.W.,National Livestock Resources Research Institute NaLIRRI | And 10 more authors.
BMC Microbiology | Year: 2010

Background. The rickettsial bacterium Ehrlichia ruminantium is the causative agent of heartwater, a potential zoonotic disease of ruminants transmitted by ticks of the genus Amblyomma. The disease is distributed in nearly all of sub-Saharan Africa and some islands of the Caribbean, from where it threatens the American mainland. This report describes the development of two different loop-mediated isothermal amplification (LAMP) assays for sensitive and specific detection of E. ruminantium. Results. Two sets of LAMP primers were designed from the pCS20 and sodB genes. The detection limits for each assay were 10 copies for pCS20 and 5 copies for sodB, which is at least 10 times higher than that of the conventional pCS20 PCR assay. DNA amplification was completed within 60 min. The assays detected 16 different isolates of E. ruminantium from geographically distinct countries as well as two attenuated vaccine isolates. No cross-reaction was observed with genetically related Rickettsiales, including zoonotic Ehrlichia species from the USA. LAMP detected more positive samples than conventional PCR but less than real-time PCR, when tested with field samples collected in sub-Saharan countries. Conclusions. Due to its simplicity and specificity, LAMP has the potential for use in resource-poor settings and also for active screening of E. ruminantium in both heartwater-endemic areas and regions that are at risk of contracting the disease. © 2010 Nakao et al; licensee BioMed Central Ltd. Source

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