Abstract
African animal trypanosomosis is a debilitating tsetse-transmitted parasitic disease of sub-Saharan Africa. Therapeutic and prophylactic drugs were introduced more than 50 years ago, and drug resistance is increasingly reported. In a cross-sectional study, 467 cattle were microscopically screened for trypanosomes. Samples were collected in May–July 2014 from five villages (Botao, Mungama, Zalala-Electrosul, Zalala-Madal, and Namitangurine) in Nicoadala district, Zambezia province. To evaluate treatment efficacy, trypanosome-positive animals in each village were randomly assigned to two groups, one treated with 0.5 mg/kg b.w. isometamidium (Inomidium®), the second with 3.5 mg/kg b.w. diminazene (Inomazene®). Cattle were microscopically monitored at days 0, 14, and 28 post-treatment. At day 28, trypanocides were swapped to investigate single or multiple resistance. Microscopically negative samples from the monitoring days were tested using 18S-PCR-RFLP. 22.9% (107/467) was found positive on day 0. On day 14, nine animals in Botao and seven in Mungama were positive. On day 28, in Botao, four animals from the diminazene group and four from the isometamidium group were positive. In Mungama, four animals from the diminazene group were positive on day 28. On day 42, six animals (9%) in Botao and two (9.5%) in Mungama remained positive after drug swap. No relapses occurred in Namitangurine. The 18S-PCR-RFLP consistently detected more positive than microscopy: indeed, positives reached 12, 13, and 8 in Botao and 9, 7, and 4 in Mungama, at days 14, 28, and 42, respectively. Single- and multi-drug resistance in Nicoadala district, Zambezia province, is thus here confirmed. This should be considered when choosing control options.
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References
Ahmed SK, Rahman AH, Hassan MA, Salih EM, Paone M, Cecchi G (2016) An atlas of tsetse and bovine trypanosomosis in Sudan. Parasit Vectors 9(1):194. https://doi.org/10.1186/s13071-016-1485-6
Baker N, de Koning HP, Mäser P, Horn D (2013) Drug resistance in African trypanosomiasis: the melarsoprol and pentamidine story. Trends Parasitol 29(3):110–118. https://doi.org/10.1016/j.pt.2012.12.005
Barret MP, Burchmore RJ, Stich AS, Lazzari JO, Frasch AC, Cazulo JJ, Krishna S (2003) The trypanosomiases. Lancet 362(9394):1469–1480. https://doi.org/10.1016/S0140-6736(03)14694-6
Büscher P, Cecchi G, Jamonneau V, Priotto G (2017) Human African trypanosomiasis. Lancet 390(10110):2397–2409. https://doi.org/10.1016/S0140-6736(17)31510-6
Cannon RM, Roe RT (1982) Livestock disease surveys: a field manual for veterinarians. Australian Government Publishing Service, Canberra
Cecchi G, Paone M, Feldmann U, Vreysen MJ, Diall O, Mattioli RC (2014) Assembling a geospatial database of tsetse-transmitted animal trypanosomosis for Africa. Parasit Vectors 7(1):39. https://doi.org/10.1186/1756-3305-7-39
Chitanga S, Marcotty T, Namangala R, Van den Bossche P, Van Den Abbeele J, Delespaux V (2011) High prevalence of drug resistance in animal trypanosomes without a history of drug exposure. PLoS Negl Trop Dis 5(12):e1454. https://doi.org/10.1371/journal.pntd.0001454
Clausen PH, Sidibe I, Kabore I, Bauer B (1992) Development of multiple drug resistance of Trypanosoma congolense in Zebu cattle under high natural tsetse fly challenge in the pastoral zone of Samorogouan, Burkina Faso. Acta Trop 51:229–236
Clausen PH, Wiemann A, Patzelt R, Kakaire D, Poetzsch C, Peregrine A, Mehlitz D (1998) Use of a PCR assay for the specific and sensitive detection of Trypanosoma spp. in naturally infected dairy cattle in peri-urban Kampala, Uganda. Ann N Y Acad Sci 849(1):21–32. https://doi.org/10.1111/j.1749-6632.1998.tb11029.x
Clausen PH, Bauer B, Zessin KH, Diall O, Bocoum Z, Sidibe I, Affognon H, Waibel H, Grace D, Randolph T (2010) Preventing and containing trypanocide resistance in the cotton zone of West Africa. Transbound Emerg Dis 57:28–32
Cox AP, Tosas O, Tilley A, Picozzi K, Coleman P, Hide G, Welburn SC (2010) Constraints to estimating the prevalence of trypanosome infections in East African zebu cattle. Parasit Vectors 3(1):82. https://doi.org/10.1186/1756-3305-3-82
Dávila AM, Majiwa PA, Grisard EC, Aksoy S, Melville SE (2003) Comparative genomics to uncover the secrets of tsetse and livestock-infective trypanosomes. Trends Parasitol 19(10):436–439. https://doi.org/10.1016/S1471-4922(03)00196-X
De Koning HP, Anderson LF, Stewart M, Burchmore RJ, Wallace LJ, Barrett MP (2004) The trypanocides diminazene aceturate is accumulated predominantly through the TbAT1 purine transporter: additional insights on diamidine resistance in African trypanosomes. Antimicrob Agents Chemother 48(5):1515–1519. https://doi.org/10.1128/AAC.48.5.1515-1519.2004
Delespaux V, De Koning HP (2007) Drugs and drug resistance in African trypanosomiasis. Drug Resist Update 10:30–50
Delespaux V, Chitanga S, Geysen D, Goethals A, Van den Bossche P, Geerts S (2006) SSCP analysis of the P2 purine transporter TcoAT1 gene of Trypanosoma congolense leads to a simple PCR-RFLP test allowing the rapid identification of diminazene resistant stocks. Acta Trop 100(1-2):96–102. https://doi.org/10.1016/j.actatropica.2006.10.001
Delespaux V, Dinka H, Masumu J, Van den Bossche P, Geerts S (2008) Five fold increase in Trypanosoma congolense isolates resistant to diminazene aceturate over a seven-year period in Eastern Zambia. Drug Resist Update 11:205–209
Desquenes M, Davila AM (2002) Applications of PCR-based tools for detection and identification of animal trypanosomes: a review and perspectives. Vet Parasitol 109(3-4):213–231. https://doi.org/10.1016/S0304-4017(02)00270-4
Diall O, Cecchi G, Wanda G, Argilés-Herrero R, Vreysen MJ, Cattoli G, Viljoen GJ, Mattioli R, Bouyer J (2017) Developing a progressive control pathway for African animal trypanosomosis. Trends Parasitol 33(7):499–509. https://doi.org/10.1016/j.pt.2017.02.005
Eisler M, Brandt J, Bauer B, Clausen P, Delespaux V, Holmes P, Ilemobade A, Machila N, Mbwambo H, McDermott J, Mehlitz D, Murilla G, Ndung’u J, Peregrine A, Sidibe I, Sinyangwe L, Geerts S (2001) Standardized tests in mice and cattle for detection of drug resistance in tsetse transmitted trypanosomes of African domestic cattle. Vet Parasitol 97(3):171–182. https://doi.org/10.1016/S0304-4017(01)00415-0
Franco JR et al (2017) Monitoring the elimination of human African trypanosomiasis: update to 2014. PLoS Negl Trop Dis 11(5):e00005585
Geerts S, Holmes PH (1998) Drug management and parasite resistance in bovine trypanosomiasis in Africa. Drug management and parasite resistance in bovine trypanosomiasis in Africa. PAAT Technical and Scientific Series. FAO
Geerts S, Holmes PH, Eisler MC, Diall O (2001) African bovine trypanosomiasis: the problem of drug resistance. Trends Parasitol 17(1):25–28. https://doi.org/10.1016/S1471-4922(00)01827-4
Geysen D, Delespaux V, Geerts S (2003) PCR–RFLP using Ssu-rDNA amplification as an easy method for species-specific diagnosis of Trypanosoma species in cattle. Vet Parasitol 110:171–180
Jamal SAJ, Sigauque I, Macuamule C, Neves L, Penzhorn BL, Marcotty T, Van den Bossche P (2005) The susceptibility of Trypanosoma congolense isolated in Zambezia Province, Mozambique, to isometamidium chloride, diminazene aceturate and homidium chloride. Onderstepoort J Vet Res 72(4):333–338
Laohasinnarong D, Thekisoe OM, Malele I, Namangala B, Ishii A, Goto Y, Kawazu SI, Sugimoto C, Inoue N (2011) Prevalence of Trypanosoma sp. in cattle from Tanzania estimated by conventional PCR and loop-mediated isothermal amplification (LAMP). Parasitol Res 109(6):1735–1739. https://doi.org/10.1007/s00436-011-2513-2
Macucule PA (2014) Diagnosis and mapping of diminazene aceturate resistance in Trypanosoma congolense, Broden 1904, strains circulating in cattle in Matutuíne district, Mozambique. Dissertation, University of Pretoria
Malele II, Magwisha HB, Nyingilili HS, Mamiro KA, Rukambile EJ, Daffa JW, Lyaruu EA, Kapange LA, Kasilagila G, Lwitiko N, Msami HM, Kimbita EN (2011) Multiple Trypanosoma infections are common amongst Glossina species in the new farming areas of Rufiji district, Tanzania. Parasit Vectors 4(1):217. https://doi.org/10.1186/1756-3305-4-217
Marcotty T, Simukoko H, Berkvens D, Vercruysse J, Praet N, Van den Bossche P (2008) Evaluating the use of packed cell volume as an indicator of trypanosomal infections in cattle in eastern Zambia. Prev Vet Med 87(3-4):288–300. https://doi.org/10.1016/j.prevetmed.2008.05.002
Matovu E, Stewart ML, Geiser F, Brun R, Mäser P, Wallace LJ, Burchmore RJ, Enyaru JC, Barret MP, Kaminsky R, Seebeck T, De Koning HP (2003) Mechanisms of arsenical and diamidine uptake and resistance in Trypanosoma brucei. Eukaryot Cell 2(5):1003–1008. https://doi.org/10.1128/EC.2.5.1003-1008.2003
Mbewe NJ, Namangala B, Sitali L, Vorster I, Michelo C (2015) Prevalence of pathogenic trypanosomes in anaemic cattle from trypanosomosis challenged areas of Itezhi-tezhi district in central Zambia. Parasit Vectors 8(1):638. https://doi.org/10.1186/s13071-015-1260-0
McDermott J, Woitag T, Sidibé I, Bauer B, Diarra B, Oue’draogo D, Kamuanga M, Peregrine A, Eisler M, Zessin KH, Mehlitz D, Clausen PH (2003) Field studies of drug-resistance cattle trypanosomes in Ke’ne’dougou Province, Burkina Faso. Acta Trop 86(1):93–103. https://doi.org/10.1016/S0001-706X(03)00019-6
Miruk A, Hagos A, Yacob HT, Asnake F, Basu AK (2008) Prevalence of bovine trypanosomosis and trypanocidal drug sensitivity studies on Trypanosoma congolense in Wolyta and Dawero zones of southern Ethiopia. Vet Parasitol 152(1-2):141–147. https://doi.org/10.1016/j.vetpar.2007.12.007
Mulugeta W, Wilkes J, Mulatu W, Majiwa PA, Masake R, Peregrine AS (1997) Long-term occurrence of Trypanosoma congolense resistant to diminazene, isometamidium and homidium in cattle at Ghibe, Ethiopia. Acta Trop 64(3-4):205–217. https://doi.org/10.1016/S0001-706X(96)00645-6
Munday J, López K, Eze A, Delespaux V, Van Den Abbeele J, Rowan T, Barret M, Morrison L, De Koning H (2013) Functional expression of TcoAT1 reveals it to be a P1-type nucleoside transporter with no capacity for diminazene uptake. Int J Parasitol Drugs Drug Resist 3:69–76. https://doi.org/10.1016/j.ijpddr.2013.01.004
Mungube E, Vitouley H, Diall O, Cudjoe E, Boucoum Z, Diarra B, Sanogo Y, Randolph T, Bauer B, Zessin K, Clausen PH (2012a) Detection of multiple drug-resistant Trypanosoma congolense populations in village cattle of south-east Mali. Parasit Vectors 5(1):155. https://doi.org/10.1186/1756-3305-5-155
Mungube E, Diall O, Baumann M, Sanogo Y, Randolph T, Bauer B, Hoppenheit A, Hinney B, Maiga B, Zessin K, Clausen PH (2012b) Best-bet integrated strategies for containing drug-resistant trypanosomes in cattle. Parasit Vectors 5(1):164. https://doi.org/10.1186/1756-3305-5-164
Murray M, Murray PK, McIntyre WI (1977) An improved parasitological tecnique for diagnosis of African trypanosomiasis. Trans R Soc Trop Med Hyg 71(4):325–326. https://doi.org/10.1016/0035-9203(77)90110-9
Mwandiringana E, Gori E, Nyengerai T, Chidzwondo F (2012) Polymerase chain reaction (PCR) detection of mixed trypanosome infection and blood meal origin in field-captured tsetse flies from Zambia. Afr J Biotechnol 11:14490–14497
Paris J, Murray M, McOdimba F (1982) A comparative evaluation of the parasitological techniques currently available for the diagnosis of African trypanosomiasis in cattle. Acta Trop 39(4):307–316
Rafael AAG (1959) Ensaios com o Pro-salt de Antricide R.F. e Berenil. Separata do Boletim No 117
RTTCP (2000) An update of the distribution of bovine trypanosomosis in Mozambique (1986–1998)
Shah-Fischer M, Say RR (1989) Manual of tropical veterinary parasitology. U.K. The Technical Centre for Agriculture and Rural Cooperation
Shaw APM, Cecchi G, Wint GRW, Mattioli RC, Robinson TP (2014) Mapping the economic benefits to livestock keepers from intervening against bovine trypanosomosis in Eastern Africa. Prev Vet Med 113(2):197–210. https://doi.org/10.1016/j.prevetmed.2013.10.024
Sigauque I, Van den Bossche P, Moiana M, Jamal S, Neves L (2000) The distribution of tsetse (Diptera: Glossinidae) and bovine trypanosomosis in the Matutuine District, Maputo Province, Mozambique. Onderstepoort J Vet Res 67(3):167–172
Silva JM (1959) Aspectos das trypanossomiases animais em Moçambique – Revisão da sua terapêutica. Separata do Boletim No115
Simo G, Fongho P, Farikou O, Ndjeuto-Tchouli PIN, Tchouomene-Labou J, Njiokou F, Asonganyi T (2015) Trypanosome infection rates in tsetse flies in the “silent” sleeping sickness focus of Bafia in the centre region in Cameroon. Parasit Vectors 8(1):528. https://doi.org/10.1186/s13071-015-1156-z
Simukoko H, Marcotty T, Phiri I, Geysen D, Vercruysse J, Van den Bossche P (2007) The comparative role of cattle, goats and pigs in the epidemiology of livestock trypanosomiasis on the plateau of eastern Zambia. Vet Parasitol 147(3-4):231–238. https://doi.org/10.1016/j.vetpar.2007.04.005
Sinyangwe L, Delespaux V, Brandt J, Geerts S, Mubanga J, Machila N, Holmes PH, Eisler MC (2004) Trypanocidal drug resistance in eastern province of Zambia. Vet Parasitol 119(2-3):125–135. https://doi.org/10.1016/j.vetpar.2003.11.007
Specht EJK (2008) Prevalence of bovine trypanosomosis in Central Mozambique from 2002 to 2005. Onderstepoort J Vet Res 75(1):73–81
Sutherland IA, Holmes PH (1993) Alterations in drug transport in resistant Trypanosoma congolense. Acta Trop 54(3-4):271–278. https://doi.org/10.1016/0001-706X(93)90099-W
Sutherland IA, Peregrine AS, Lonsdale-Eccles JD, Holmes PH (1991) Reduced accumulation of isometamidium by drug-resistant Trypanosoma congolense. Parasitology 103(02):245–251. https://doi.org/10.1017/S0031182000059527
Sutherland IA, Mounsey A, Holmes PH (1992) Transport of isometamidium (Samorin) by drug-resistant and drug-sensitive Trypanosoma congolense. Parasitology 104(03):461–467. https://doi.org/10.1017/S0031182000063721
Tchamdja E, Kulo AE, Vitouley HS, Batawui K, Bankolé AA, Adomefa K, Cecchi G, Hoppenheit A, Clausen PH, De Deken R, Van Den Abbeele J, Marcotty T, Delespaux V (2017) Cattle breeding, trypanosomosis prevalence and drug resistance in Northern Togo. Vet Parasitol 236:86–92. https://doi.org/10.1016/j.vetpar.2017.02.008
Van den Bossche P (2001) Some general aspects of the distribution and epidemiology of bovine trypanosomosis in southern Africa. Int J Parasitol 31(5-6):592–598. https://doi.org/10.1016/S0020-7519(01)00146-1
Acknowledgements
We would like to acknowledge the European Union for financing the study through the EU funded TRYRAC project (TRYRAC/DCI-FOOD/2011/279-754), the Biotechnology Center - Eduardo Mondlane University and the University of Pretoria for all the laboratory assistance.
FAO assistance to this study was provided in the framework of the Programme Against African Trypanosomosis (PAAT), and supported by the Government of Italy (Project ‘Improving food security in sub-Saharan Africa by supporting the progressive reduction of tsetse-transmitted trypanosomosis in the framework of the NEPAD’, codes GTFS/RAF/474/ITA and GCP/RAF/502/ITA).
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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed and all procedures performed in studies involving animals were in accordance with the ethical standards of Biotechnology Centre—Eduardo Mondlane University and the practice at which the study was conducted. Permit number CBUEM/COMETH_0014/2014 issued on 17 March 2014.
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Mulandane, F.C., Fafetine, J., Van Den Abbeele, J. et al. Resistance to trypanocidal drugs in cattle populations of Zambezia Province, Mozambique. Parasitol Res 117, 429–436 (2018). https://doi.org/10.1007/s00436-017-5718-1
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DOI: https://doi.org/10.1007/s00436-017-5718-1
Keywords
- African animal trypanosomosis
- Chemo-resistance
- Trypanocides
- Block treatment
- PCR-RFLP