Molecular Studies of Gastrointestinal Strongyle Nematodes in Migratory, Resident, and Sedentary Plains Zebras (Equus quagga) in Kenya
Gastrointestinal Strongyle Nematodes in Zebras
Keywords:
Molecular identity, phylogeny, strongyles, plains zebrasAbstract
The molecular identity of gastrointestinal (GI) strongyle larvae recovered from faecal cultures from plains zebras in Kenya was determined using molecular tools. Internal transcribed spacer (ITS) ribosomal DNA (rDNA) extracted from the larvae were amplified using Polymerase Chain Reaction (PCR), sequenced for identification, and compared to sequences in the GenBank to determine their phylogeny. Sixteen sequences were obtained and identified as Cyathostomum montgomeryi, Cylicostephanus minutus, Poteriostomum imparidentatum, Triodontophorus nipponicus and Strongylus vulgaris. The genetic identity of P. imparidentatum, T. nipponicus and S. vulgaris from plains zebras in Kenya are reported for the first time in this study. The 16 sequences clustered into 5 clades according to the 5 genera of nematodes identified. The clade having T. nipponicus was placed as a sister to the Cyathostominae but was very distinct from the clade having S. vulgaris. The close clustering of T. nipponicus to the Cyathostominae supports previous suggestions that it belongs to this subfamily rather than Strongylinae. Five sequences of C. montgomeryi clustered closely with four sequences of C. montgomeryi in the GenBank isolated from zebras in Kenya. The other five sequences were evolutionary distinct. Similarly, two of the Cy. minutus sequences clustered with Cy. minutus sequences from zebras in Kenya, while the other one, was distinct. These results suggest intra-species genetic polymorphism among the C. montgomeryi and Cy. minutus isolates. The Poteriostomum imparidentatum, T. nipponicus, and S. vulgaris sequences were distinct from sequences found in the GenBank. This study contributes to the scanty but growing literature on equine strongyle genetics in zebras.
References
Burgess CGS, Bartley Y, Redman E, Skuce PJ, Nath M, Whitelaw F, et al. A survey of the trichostrongylid nematode species present on UK sheep farms and associated anthelmintic control practices. Vet Parasitol 2012;189:299–307. https://doi.org/10.1016/j.vetpar.2012.04.009.
Petney TN, Andrews RH. Multiparasite communities in animals and humans: frequency, structure and pathogenic significance. Int J Parasitol 1998;28:377–93. https://doi.org/10.1016/s0020-7519(97)00189-6.
Gulland FM, Fox M. Epidemiology of nematode infections of Soay sheep (Ovis aries L.) on St Kilda. Parasitology 1992;105 (Pt 3):481–92. https://doi.org/10.1017/s0031182000074667.
Harvell CD, Mitchell CE, Ward JR, Altizer S, Dobson AP, Ostfeld RS, et al. Climate warming and disease risks for terrestrial and marine biota. Science 2002;296:2158–62. https://doi.org/10.1126/science.1063699.
Michel AL, Bengis RG. The African buffalo: A villain for inter-species spread of infectious diseases in southern Africa. Onderstepoort Journal of Veterinary Research 2012;79:26–30.
Lichtenfels JR, Kharchenko VA, Dvojnos GM. Illustrated identification keys to strongylid parasites (strongylidae: Nematoda) of horses, zebras and asses (Equidae). Veterinary Parasitology 2008;156:4–161. https://doi.org/10.1016/j.vetpar.2008.04.026.
Tombak KJ, Hansen CB, Kinsella JM, Pansu J, Pringle RM, Rubenstein DI. The gastrointestinal nematodes of plains and Grevy's zebras: Phylogenetic relationships and host specificity. International Journal for Parasitology: Parasites and Wildlife 2021;16:228–35. https://doi.org/10.1016/j.ijppaw.2021.10.007.
Krecek RC, Malan FS, Reinecke RK, de Vos V. Nematode parasites from Burchell's zebras in South Africa. J Wildl Dis 1987;23:404–11. https://doi.org/10.7589/0090-3558-23.3.404.
Mitchell CJ, O'Sullivan CM, Pinloche E, Wilkinson T, Morphew RM, MCewan NR. Using next-generation sequencing to determine diversity of horse intestinal worms: identifying the equine 'nemabiome.' J Equine Sci 2019;30:1–5. https://doi.org/10.1294/jes.30.1.
Gao Y, Zhang Y, Yang X, Qiu J-H, Duan H, Xu W-W, et al. Mitochondrial DNA evidence supports the hypothesis that Triodontophorus species belong to Cyathostominae. Frontiers in Microbiology 2017;8.
Krecek RC, Reinecke RK, Kriek NJP, Horak IG, Malan FS. Helminth parasites of Cape mountain zebras from Cape Province, South Africa. Journal of Wildlife Diseases 1994;30:277–80. https://doi.org/10.7589/0090-3558-30.2.277.
Wambwa EN, Ogara WO, Mudakha D. A comparative study of gastrointestinal parasites between ranched and free ranging Burchell's zebra (Equus burchelli antiquorum) in Isiolo district, Kenya. J Vet Sci 2004;5:215–20.
Matthews JB. Anthelmintic resistance in equine nematodes. Int J Parasitol Drugs Drug Resist 2014;4:310–5. https://doi.org/10.1016/j.ijpddr.2014.10.003.
Hamed MI, El-Allawy TA, Hassnein E. Prevalence and anthelmintic resistance of Strongyle infection of donkeys in El-Wadi El-Gadid, Egypt. Journal of Advanced Veterinary Research 2019;9:144–50.
Oakenfull EA, Clegg JB. Phylogenetic relationships within the genus Equus and the evolution of alpha and theta globin genes. J Mol Evol 1998;47:772–83. https://doi.org/10.1007/pl00006436.
Matthee S, Krecek RC, McGeoch MA. A comparison of the intestinal helminth communities of Equidae in Southern Africa. J Parasitol 2004;90:1263–73. https://doi.org/10.1645/GE-3353.
Tolliver SC, Lyons ET, Drudge JH. Species of small strongyles and other internal parasites recovered from donkeys at necropsy in Kentucky. Proceedings of the Helminthological Society of Washington (USA) 1985;52:260–5.
Barker DC. Molecular approaches to DNA diagnosis. Parasitology 1989;99 Suppl:S125-146. https://doi.org/10.1017/s0031182000083463.
Wilson SM. Nucleic acid techniques and the detection of parasitic diseases. Parasitol Today 1991;7:255–9. https://doi.org/10.1016/0169-4758(91)90243-h.
Mullis K, Faloona F, Scharf S, Saiki R, Horn G, Erlich H. Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. Cold Spring Harb Symp Quant Biol 1986;51 Pt 1:263–73. https://doi.org/10.1101/sqb.1986.051.01.032.
Hung GC, Gasser RB, Beveridge I, Chilton NB. Species-specific amplification by PCR of ribosomal DNA from some equine strongyles. Parasitology 1999;119 (Pt 1):69–80. https://doi.org/10.1017/s0031182099004497.
Gasser RB, Bott NJ, Chilton NB, Hunt P, Beveridge I. Toward practical, DNA-based diagnostic methods for parasitic nematodes of livestock--bionomic and biotechnological implications. Biotechnol Adv 2008;26:325–34. https://doi.org/10.1016/j.biotechadv.2008.03.003.
Maina LGM, Maingi N, Ng'ang'a CJ, Waruiru RM, Gakuya F. Diversity, prevalence, and intensity of gastrointestinal helminth infections in migratory, resident, and sedentary plains zebras (Equus quagga) in Masai Mara National Reserve and Lake Nakuru National Park, Kenya. Vet Parasitol Reg Stud Reports 2022;33:100750. https://doi.org/10.1016/j.vprsr.2022.100750.
Great Britain Ministry of Agriculture, Fisheries and Food. Manual of Veterinary parasitological laboratory techniques. H. M. Stationery Office; 1986.
Archie EA, Ezenwa VO. Population genetic structure and history of a generalist parasite infecting multiple sympatric host species. Int J Parasitol 2011;41:89–98. https://doi.org/10.1016/j.ijpara.2010.07.014.
McLean ER, Kinsella JM, Chiyo P, Obanda V, Moss C, Archie EA. Genetic identification of five strongyle nematode parasites in wild african elephants (Loxodonta africana). J Wildl Dis 2012;48:707–16. https://doi.org/10.7589/0090-3558-48.3.707.
Edgar RC. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research 2004;32:1792–7. https://doi.org/10.1093/nar/gkh340.
Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 2018;35:1547–9. https://doi.org/10.1093/molbev/msy096.
Tamura K. Estimation of the number of nucleotide substitutions when there are strong transition-transversion and G+C-content biases. Mol Biol Evol 1992;9:678–87. https://doi.org/10.1093/oxfordjournals.molbev.a040752.
Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 1985;39:783–91. https://doi.org/10.2307/2408678.
Lichtenfels JR. Helminths of domestic equids. Illustrated keys to genera and species with emphasis on North American forms. Proceedings of the Helminthological Society of Washington 1975;42:1–92.
Kharchenko V, Lichtenfels J, Krecek R. Cyathostomum montgomeryi and its place in the Cyathostominae (Nematoda : Strongylidae). Comparative Parasitology 2001;68:97–102.
Scialdo-Krecek RC. Studies on the parasites of zebras. 1. Nematodes of the Burchell's zebra in the Kruger National Park. Onderstepoort J Vet Res 1983;50:111–4.
Kuzmina T, Kharchenko V, Zvegintsova N, Zhang L, Liu J. Strongylids (Nematoda: Strongylidae) in two zebra species from the “Askania-Nova” Biosphere Reserve, Ukraine: biodiversity and parasite community structure. Helminthologia 2013;50:172–80. https://doi.org/10.2478/s11687-013-0128-0.
Dvojnos GM, Harcenko VA. Morphology and differential diagnostics of parasitic larvae of some Strongylidae (Nematoda) of horses. Angew Parasitol 1990;31:15–28.
Chilton NB, Gasser RB, Beveridge I. Differences in a ribosomal DNA sequence of morphologically indistinguishable species within the Hypodontus macropi complex (Nematoda: Strongyloidea). International Journal for Parasitology 1995;25:647–51. https://doi.org/10.1016/0020-7519(94)00171-J.
Hung G-C, Jacobs DE, Krecek RC, Gasser RB, Chilton NB. Strongylus asini (Nematoda, Strongyloidea): Genetic relationships with other strongylus species determined by ribosomal DNA. International Journal for Parasitology 1996;26:1407–11. https://doi.org/10.1016/S0020-7519(96)00136-1.
Newton LA, Chilton NB, Beveridge I, Hoste H, Nansen P, Gasser RB. Genetic markers for strongylid nematodes of livestock defined by PCR-based restriction analysis of spacer rDNA. Acta Trop 1998;69:1–15. https://doi.org/10.1016/s0001-706x(97)00105-8.
Gasser RB, Newton SE. Genomic and genetic research on bursate nematodes: significance, implications and prospects. International Journal for Parasitology 2000;30:509–34. https://doi.org/10.1016/S0020-7519(00)00021-7.
Blouin MS. Molecular prospecting for cryptic species of nematodes: mitochondrial DNA versus internal transcribed spacer. International Journal for Parasitology 2002;32:527–31. https://doi.org/10.1016/S0020-7519(01)00357-5.
Denver DR, Wilhelm LJ, Howe DK, Gafner K, Dolan PC, Baer CF. Variation in base-substitution mutation in experimental and natural lineages of Caenorhabditis nematodes. Genome Biology and Evolution 2012;4:513–22. https://doi.org/10.1093/gbe/evs028.
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Copyright (c) 2023 Linda G.M. Maina, Ndichu Maingi, Chege J. Ng'ang'a, Robert M. Waruiru, Francis Gakuya, Esther G. Kanduma
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