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      Molecular identification of diminazene aceturate resistant trypanosomes in tsetse flies from Yoko in the Centre region of Cameroon and its epidemiological implications

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          Abstract

          African animal trypanosomiases are caused by trypanosomes cyclically or mechanically transmitted by tsetse and other biting flies. Although molecular tools have been developed to identify drug-resistant trypanosomes in mammals, little or no investigation on drug-resistance has been undertaken on trypanosomes harbored by tsetse flies. Moreover, no data on mechanical vectors of African trypanosomes is available in most endemic areas of Cameroon. This study was designed to update our knowledge on the cyclical and mechanical vectors of African trypanosomes, and using molecular tools to identify different trypanosome species as well as diminazene aceturate resistant trypanosomes in tsetse flies trapped at Yoko in the Centre region of Cameroon.

          For this study, traps were used to catch tsetse and mechanical vectors of African trypanosomes. The flies trapped were counted and identified by sex and species. DNA was extracted from tsetse and species-specific primers were used to identify different trypanosome species. PCR-RFLP was used to detect diminazene aceturate resistant strains of Trypanosoma congolense.

          In all, 454 flies comprising 168 (37%) Tabanus spp., 71 (15.6%) Stomoxys spp. and 215 (47.4%) tsetse fly ( i.e. 107 (49.8%) Glossina fusca congolensis, 71 (33%) Glossina fusca fusca and 37 (17.2%) Glossina palpalis palpalis) were trapped. Trypanosome infections were identified in 12.6% (27/215) of tsetse flies: 13 in G. f. congolensis, 6 in G. p. palpalis and 5 in G. f. fusca. From 24 T. congolense positive samples, PCR-RFLP was successful on 37.5% of the samples. Four samples (16.2%) harbored T. congolense strains that were resistant to diminazene aceturate while the remaining samples had drug-sensitive strains.

          These results show for the first time the applicability of molecular tools for the identification of drug-resistant trypanosomes in tsetse. They revealed the existence of diminazene aceturate resistant strains of T. congolense in the tsetse-infested area of Yoko in the Centre region of Cameroon. Detection of drug-resistant trypanosomes in tsetse may enable scientists to map with accuracy specific areas where these parasites are transmitted. With such mapping, control strategies against African trypanosomiases could be improved by adapting control measures according to drug resistance distribution.

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          Detection of Trypanosoma congolense and Trypanosoma brucei subspecies by DNA amplification using the polymerase chain reaction.

          The nuclear DNA of Trypanosoma congolense contains a family of highly conserved 369 base pair (bp) repeats. The sequences of three cloned copies of these repeats were determined. An unrelated family of 177 bp repeats has previously been shown to occur in the nuclear DNA of Trypanosoma brucei brucei (Sloof et al. 1983a). Oligonucleotides were synthesized which prime the specific amplification of each of these repetitive DNAs by the polymerase chain reaction (PCR). Amplification of 10% of the DNA in a single parasite of T. congolense or T. brucei spp. produced sufficient amplified product to be visible as a band in an agarose gel stained with ethidium bromide. This level of detection, which does not depend on the use of radioactivity, is about 100 times more sensitive than previous detection methods based on radioactive DNA probes. The oligonucleotides did not prime the amplification of DNA sequences in other trypanosome species nor in Leishmania, mouse or human DNAs. Amplification of DNA from the blood of animals infected with T. congolense and/or T. brucei spp. permitted the identification of parasite levels far below that detectable by microscopic inspection. Since PCR amplification can be conducted on a large number of samples simultaneously, it is ideally suited for large-scale studies on the prevalence of African trypanosomes in both mammalian blood and insect vectors.
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            Tsetse flies: their biology and control using area-wide integrated pest management approaches.

            Tsetse flies are the cyclical vectors of trypanosomes, the causative agents of 'sleeping sickness' or human African trypanosomosis (HAT) in humans and 'nagana' or African animal trypanosomosis (AAT) in livestock in Sub-saharan Africa. Many consider HAT as one of the major neglected tropical diseases and AAT as the single greatest health constraint to increased livestock production. This review provides some background information on the taxonomy of tsetse flies, their unique way of reproduction (adenotrophic viviparity) making the adult stage the only one easily accessible for control, and how their ecological affinities, their distribution and population dynamics influence and dictate control efforts. The paper likewise reviews four control tactics (sequential aerosol technique, stationary attractive devices, live bait technique and the sterile insect technique) that are currently accepted as friendly to the environment, and describes their limitations and advantages and how they can best be put to practise in an IPM context. The paper discusses the different strategies for tsetse control i.e. localised versus area-wide and focusses thereafter on the principles of area-wide integrated pest management (AW-IPM) and the phased-conditional approach with the tsetse project in Senegal as a recent example. We argue that sustainable tsetse-free zones can be created on Africa mainland provided certain managerial and technical prerequisites are in place. Copyright © 2012 International Atomic Energy Agency. Published by Elsevier Inc. All rights reserved.
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              Developing a Progressive Control Pathway for African Animal Trypanosomosis.

              Progressive control pathways (PCPs) are stepwise approaches for the reduction, elimination, and eradication of human and animal diseases. They provide systematic frameworks for planning and evaluating interventions. Here we outline a PCP for tsetse-transmitted animal trypanosomosis, the scourge of poor livestock keepers in tropical Africa. Initial PCP stages focus on the establishment of national coordination structures, engagement of stakeholders, development of technical capacities, data collection and management, and pilot field interventions. The intermediate stage aims at a sustainable and economically profitable reduction of disease burden, while higher stages target elimination. The mixed-record of success and failure in past efforts against African animal trypanosomosis (AAT) makes the development of this PCP a high priority.
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                Author and article information

                Contributors
                Journal
                Parasite Epidemiol Control
                Parasite Epidemiol Control
                Parasite Epidemiology and Control
                Elsevier
                2405-6731
                07 January 2020
                May 2020
                07 January 2020
                : 9
                : e00135
                Affiliations
                [a ]Molecular Parasitology and Entomology Unit, Department of Biochemistry, Faculty of Science, University of Dschang, PO Box 67, Dschang, Cameroon
                [b ]Mission Spéciale d'Eradication des Glossines, Division Régionale tsetse Adamaoua, PO Box 263, Ngaoundéré, Cameroon
                [c ]UMR 177, IRD-CIRAD, CIRAD TA A-17/G, Campus International de Baillarguet, Montpellier Cedex 5, France
                Author notes
                [* ]Corresponding author. gustave.simo@ 123456univ-dschang.org
                Article
                S2405-6731(20)30004-0 e00135
                10.1016/j.parepi.2020.e00135
                6957779
                31956704
                06436bd8-5c52-4f19-a0b2-5ee77c4590d9
                © 2020 Published by Elsevier Ltd on behalf of World Federation of Parasitologists.

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 12 August 2019
                : 3 January 2020
                : 3 January 2020
                Categories
                Original Research article

                animal african trypanosomiasis,tsetse fly,trypanosomes,drug resistance,pcr-rflp

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