FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Parasit Vectors / Isolation, cultivation and molecular characterization of a n...
Parasites & vectors2016; 9(1); 481; doi: 10.1186/s13071-016-1755-3

Isolation, cultivation and molecular characterization of a new Trypanosoma equiperdum strain in Mongolia.

Abstract: Trypanosoma equiperdum causes dourine via sexual transmission in Equidae. T. equiperdum is classified under the subgenus Trypanozoon along with the T. brucei sspp. and T. evansi; however, the species classification of Trypanozoon remains a controversial topic due to the limited number of T. equiperdum reference strains. In addition, it is possible that some were misclassified T. evansi strains. Thus, there is a strong need for a new T. equiperdum strain directly isolated from the genital mucosa of a horse with a clinically- and parasitologically-confirmed dourine infection. Trypanosomes isolated from the urethral tract of a stallion with suspected dourine, were directly cultivated using soft agarose media at 37 °C in 5 % CO2. For molecular characterization, 18S ribosomal RNA (rRNA) gene, the internal transcribed spacer (ITS) and 8 maxicircle DNA regions were amplified by a PCR and their sequences were determined. To analyze the ratio of the kinetoplastic/akinetoplastic population, the kinetoplasts and the nuclei of trypanosomes were subjected to Hoechst staining and observed by fluorescence microscopy. In addition to the clinical symptoms and the molecular diagnosis, this stallion was definitively diagnosed with dourine by the detection of trypanosomes in the urethral mucosa. These results strongly suggested that the isolated trypanosome was true T. equiperdum. T. equiperdum isolated from the urethral tract was adapted in vitro using soft agarose media. Based on the results of a phylogenetic analysis of 18S rRNA and ITS, this T. equiperdum isolate was classified into the Trypanozoon clade. In a PCR of the maxicircle DNA region, only NADH-dehydrogenase subunits 4 and 5 was amplified. Clear kinetoplasts were observed in most of the T. equiperdum isolates. In contrast, most culture-adapted T. equiperdum were of the akinetoplastic form. We concluded that our isolated trypanosome was the first confirmed case of T. equiperdum in Mongolia and named it "T. equiperdum IVM-t1". T. equiperdum IVM-t1 was well adapted and propagated in soft agarose media, which indicates that this culture method is useful for isolation of T. equiperdum from horses with dourine.
Get Full Text
Publication Date: 2016-08-31 PubMed ID: 27580944PubMed Central: PMC5007690DOI: 10.1186/s13071-016-1755-3Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Case Reports
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The study introduces the isolation, cultivation, and molecular characterization of a new strain of Trypanosoma equiperdum found in Mongolia, originating from a horse with a confirmed case of dourine, a sexually transmitted disease caused by T. equiperdum.

Introduction and Methodology

  • The research seeks to clarify uncertainties surrounding the species classification of Trypanozoon, under which T. equiperdum falls, along with T. brucei sspp and T. evansi. This uncertainty is due to the limited number of T. equiperdum reference strains available and the possibility of misclassified T.evansi strains.
  • The researchers isolated trypanosomes, a type of parasitic protozoa, from the urethral tract of a stallion with suspected dourine. They cultivated these samples using soft agarose media under specific conditions (37 °C in 5 % CO2).

Molecular Characterization and Analysis

  • To characterize the isolated strain molecularly, 18S ribosomal RNA (rRNA) gene, the internal transcribed spacer (ITS), and 8 maxicircle DNA regions were studied. These elements were amplified by a PCR process and their sequences were determined.
  • Microscopic examination after staining revealed the presence of kinetoplasts and nuclei in the observed trypanosomes. This was done to analyze the ratio of the kinetoplastic/akinetoplastic population of the strain.

Results

  • The molecular diagnosis, along with observed clinical symptoms, and the detection of trypanosomes in the urethral mucosa led to a confirmed diagnosis of dourine in the stallion.
  • The isolated strain was adapted in vitro with soft agarose media and based on phylogenetic analysis involving 18S rRNA and ITS, its classification into the Trypanozoon clade was confirmed.
  • PCR amplification only occurred for NADH-dehydrogenase subunits 4 and 5 in the maxicircle DNA region. The strain was further characterized by the clear presence of kinetoplasts, distinguishing it from most akinetoplastic culture-adapted T. equiperdum.

Conclusion

  • The isolated strain, named T. equiperdum IVM-t1, represents the first confirmed case of T. equiperdum in Mongolia.
  • It was able to adapt and propagate well in soft agarose media, suggesting the method’s utility for isolating T. equiperdum from horses with dourine.

Cite This Article

APA
Suganuma K, Narantsatsral S, Battur B, Yamasaki S, Otgonsuren D, Musinguzi SP, Davaasuren B, Battsetseg B, Inoue N. (2016). Isolation, cultivation and molecular characterization of a new Trypanosoma equiperdum strain in Mongolia. Parasit Vectors, 9(1), 481. https://doi.org/10.1186/s13071-016-1755-3

Publication

Parasites & vectors
ISSN: 1756-3305
NlmUniqueID: 101462774
Country: England
Language: English
Volume: 9
Issue: 1
Pages: 481
PII: 481

Researcher Affiliations

Suganuma, Keisuke
  • National Research Center for Protozoan Diseases, OIE Reference Laboratory for Surra, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, 080-8555, Japan.
Narantsatsral, Sandagdorj
  • Institute of Veterinary Medicine, Laboratory of Molecular Genetics, Mongolian University of Life Sciences, Zaisan, 17024, Ulaanbaatar, Mongolia.
Battur, Banzragch
  • Institute of Veterinary Medicine, Laboratory of Molecular Genetics, Mongolian University of Life Sciences, Zaisan, 17024, Ulaanbaatar, Mongolia.
Yamasaki, Shino
  • National Research Center for Protozoan Diseases, OIE Reference Laboratory for Surra, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, 080-8555, Japan.
Otgonsuren, Davaajav
  • Institute of Veterinary Medicine, Laboratory of Molecular Genetics, Mongolian University of Life Sciences, Zaisan, 17024, Ulaanbaatar, Mongolia.
Musinguzi, Simon Peter
  • National Research Center for Protozoan Diseases, OIE Reference Laboratory for Surra, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, 080-8555, Japan.
Davaasuren, Batdorj
  • National Research Center for Protozoan Diseases, OIE Reference Laboratory for Surra, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, 080-8555, Japan.
  • Institute of Veterinary Medicine, Laboratory of Molecular Genetics, Mongolian University of Life Sciences, Zaisan, 17024, Ulaanbaatar, Mongolia.
Battsetseg, Badgar
  • Institute of Veterinary Medicine, Laboratory of Molecular Genetics, Mongolian University of Life Sciences, Zaisan, 17024, Ulaanbaatar, Mongolia.
Inoue, Noboru
  • National Research Center for Protozoan Diseases, OIE Reference Laboratory for Surra, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, 080-8555, Japan. ircpmi@obihiro.ac.jp.

MeSH Terms

  • Animals
  • Dourine / epidemiology
  • Dourine / parasitology
  • Horse Diseases / epidemiology
  • Horse Diseases / microbiology
  • Horses
  • Male
  • Mongolia
  • Sexually Transmitted Diseases / epidemiology
  • Sexually Transmitted Diseases / parasitology
  • Sexually Transmitted Diseases / veterinary
  • Trypanosoma / genetics
  • Trypanosoma / isolation & purification

References

This article includes 27 references
  1. Brun R, Hecker H, Lun ZR. Trypanosoma evansi and T. equiperdum: distribution, biology, treatment and phylogenetic relationship (a review).. Vet Parasitol 1998 Oct;79(2):95-107.
    doi: 10.1016/S0304-4017(98)00146-0pubmed: 9806490google scholar: lookup
  2. Li FJ, Lai DH, Lukes J, Chen XG, Lun ZR. Doubts about Trypanosoma equiperdum strains classed as Trypanosoma brucei or Trypanosoma evansi.. Trends Parasitol 2006 Feb;22(2):55-6; author reply 58-9.
    doi: 10.1016/j.pt.2005.12.005pubmed: 16377246google scholar: lookup
  3. Claes F, Büscher P, Touratier L, Goddeeris BM. Trypanosoma equiperdum: master of disguise or historical mistake?. Trends Parasitol 2005 Jul;21(7):316-21.
    doi: 10.1016/j.pt.2005.05.010pubmed: 15923142google scholar: lookup
  4. Hagos A, Goddeeris BM, Yilkal K, Alemu T, Fikru R, Yacob HT, Feseha G, Claes F. Efficacy of Cymelarsan and Diminasan against Trypanosoma equiperdum infections in mice and horses.. Vet Parasitol 2010 Aug 4;171(3-4):200-6.
    doi: 10.1016/j.vetpar.2010.03.041pubmed: 20417035google scholar: lookup
  5. Pascucci I, Di Provvido A, Cammà C, Di Francesco G, Calistri P, Tittarelli M, Ferri N, Scacchia M, Caporale V. Diagnosis of dourine in outbreaks in Italy.. Vet Parasitol 2013 Mar 31;193(1-3):30-8.
    doi: 10.1016/j.vetpar.2012.12.006pubmed: 23298562google scholar: lookup
  6. Clausen PH, Chuluun S, Sodnomdarjaa R, Greiner M, Noeckler K, Staak C, Zessin KH, Schein E. A field study to estimate the prevalence of Trypanosoma equiperdum in Mongolian horses.. Vet Parasitol 2003 Jul 10;115(1):9-18.
    doi: 10.1016/S0304-4017(03)00160-2pubmed: 12860063google scholar: lookup
  7. Claes F, Ilgekbayeva GD, Verloo D, Saidouldin TS, Geerts S, Buscher P, Goddeeris BM. Comparison of serological tests for equine trypanosomosis in naturally infected horses from Kazakhstan.. Vet Parasitol 2005 Aug 10;131(3-4):221-5.
    doi: 10.1016/j.vetpar.2005.05.001pubmed: 15951112google scholar: lookup
  8. Vulpiani MP, Carvelli A, Giansante D, Iannino F, Paganico D, Ferri N. Reemergence of dourine in Italy: clinical cases in some positive horses.. J Equine Vet Sci 2013;33(6):468–74.
    doi: 10.1016/j.jevs.2012.07.010google scholar: lookup
  9. Calistri P, Narcisi V, Atzeni M, De Massis F, Tittarelli M, Mercante MT, Ruggieri E, Scacchia M. Dourine reemergence in Italy.. J Equine Vet Sci 2013;33(2):83–9.
    doi: 10.1016/j.jevs.2012.05.057google scholar: lookup
  10. Nguyen TT, Zhou M, Ruttayaporn N, Nguyen QD, Nguyen VK, Goto Y, Suzuki Y, Kawazu S, Inoue N. Diagnostic value of the recombinant tandem repeat antigen TeGM6-4r for surra in water buffaloes.. Vet Parasitol 2014 Mar 17;201(1-2):18-23.
    doi: 10.1016/j.vetpar.2014.01.009pubmed: 24524896google scholar: lookup
  11. Nguyen TT, Ruttayaporn N, Goto Y, Kawazu S, Sakurai T, Inoue N. A TeGM6-4r antigen-based immunochromatographic test (ICT) for animal trypanosomosis.. Parasitol Res 2015 Nov;114(11):4319-25.
    doi: 10.1007/s00436-015-4672-zpubmed: 26290217google scholar: lookup
  12. Desquesnes M, McLaughlin G, Zoungrana A, Dávila AM. Detection and identification of Trypanosoma of African livestock through a single PCR based on internal transcribed spacer 1 of rDNA.. Int J Parasitol 2001 May 1;31(5-6):610-4.
    doi: 10.1016/S0020-7519(01)00161-8pubmed: 11334950google scholar: lookup
  13. Thuy NT, Goto Y, Lun ZR, Kawazu S, Inoue N. Tandem repeat protein as potential diagnostic antigen for Trypanosoma evansi infection.. Parasitol Res 2012 Feb;110(2):733-9.
    doi: 10.1007/s00436-011-2632-9pubmed: 21927872google scholar: lookup
  14. Herbert WJ, Lumsden WH. Trypanosoma brucei: a rapid "matching" method for estimating the host's parasitemia.. Exp Parasitol 1976 Dec;40(3):427-31.
    doi: 10.1016/0014-4894(76)90110-7pubmed: 976425google scholar: lookup
  15. Hirumi H, Hirumi K. In vitro cultivation of Trypanosoma congolense bloodstream forms in the absence of feeder cell layers.. Parasitology 1991 Apr;102 Pt 2:225-36.
    doi: 10.1017/S0031182000062533pubmed: 1852490google scholar: lookup
  16. Sambrook J, Russell DW, Sambrook J. The condensed protocols from molecular cloning: a laboratory manual.. Cold Spring Harbor: Cold Spring Harbor Laboratory Press; 2006.
  17. Da Silva FM, Noyes H, Campaner M, Junqueira AC, Coura JR, Añez N, Shaw JJ, Stevens JR, Teixeira MM. Phylogeny, taxonomy and grouping of Trypanosoma rangeli isolates from man, triatomines and sylvatic mammals from widespread geographical origin based on SSU and ITS ribosomal sequences.. Parasitology 2004 Nov;129(Pt 5):549-61.
    doi: 10.1017/S0031182004005931pubmed: 15552400google scholar: lookup
  18. Lai DH, Hashimi H, Lun ZR, Ayala FJ, Lukes J. Adaptations of Trypanosoma brucei to gradual loss of kinetoplast DNA: Trypanosoma equiperdum and Trypanosoma evansi are petite mutants of T. brucei.. Proc Natl Acad Sci U S A 2008 Feb 12;105(6):1999-2004.
    doi: 10.1073/pnas.0711799105pmc: PMC2538871pubmed: 18245376google scholar: lookup
  19. Suganuma K, Sarwono AE, Mitsuhashi S, Jąkalski M, Okada T, Nthatisi M, Yamagishi J, Ubukata M, Inoue N. Mycophenolic Acid and Its Derivatives as Potential Chemotherapeutic Agents Targeting Inosine Monophosphate Dehydrogenase in Trypanosoma congolense.. Antimicrob Agents Chemother 2016 Jul;60(7):4391-3.
    doi: 10.1128/AAC.02816-15pmc: PMC4914658pubmed: 27139487google scholar: lookup
  20. Carruthers VB, Cross GA. High-efficiency clonal growth of bloodstream- and insect-form Trypanosoma brucei on agarose plates.. Proc Natl Acad Sci U S A 1992 Sep 15;89(18):8818-21.
    doi: 10.1073/pnas.89.18.8818pmc: PMC50012pubmed: 1528898google scholar: lookup
  21. King SRB, Gurnell J. Effects of fly disturbance on the behaviour of a population of reintroduced Przewalski horses (Equus ferus przewalskii) in Mongolia.. Appl Anim Behav Sci 2010;125(1–2):22–9.
    doi: 10.1016/j.applanim.2010.03.006google scholar: lookup
  22. Carnes J, Anupama A, Balmer O, Jackson A, Lewis M, Brown R, Cestari I, Desquesnes M, Gendrin C, Hertz-Fowler C, Imamura H, Ivens A, Kořený L, Lai DH, MacLeod A, McDermott SM, Merritt C, Monnerat S, Moon W, Myler P, Phan I, Ramasamy G, Sivam D, Lun ZR, Lukeš J, Stuart K, Schnaufer A. Genome and phylogenetic analyses of Trypanosoma evansi reveal extensive similarity to T. brucei and multiple independent origins for dyskinetoplasty.. PLoS Negl Trop Dis 2015 Jan;9(1):e3404.
    doi: 10.1371/journal.pntd.0003404pmc: PMC4288722pubmed: 25568942google scholar: lookup
  23. Sánchez E, Perrone T, Recchimuzzi G, Cardozo I, Biteau N, Aso PM, Mijares A, Baltz T, Berthier D, Balzano-Nogueira L, Gonzatti MI. Molecular characterization and classification of Trypanosoma spp. Venezuelan isolates based on microsatellite markers and kinetoplast maxicircle genes.. Parasit Vectors 2015 Oct 15;8:536.
    doi: 10.1186/s13071-015-1129-2pmc: PMC4607141pubmed: 26467019google scholar: lookup
  24. Desquesnes M, Holzmuller P, Lai DH, Dargantes A, Lun ZR, Jittaplapong S. Trypanosoma evansi and surra: a review and perspectives on origin, history, distribution, taxonomy, morphology, hosts, and pathogenic effects.. Biomed Res Int 2013;2013:194176.
    pmc: PMC3760267pubmed: 24024184doi: 10.1155/2013/194176google scholar: lookup
  25. Schnaufer A, Domingo GJ, Stuart K. Natural and induced dyskinetoplastic trypanosomatids: how to live without mitochondrial DNA.. Int J Parasitol 2002 Aug;32(9):1071-84.
    doi: 10.1016/S0020-7519(02)00020-6pubmed: 12117490google scholar: lookup
  26. Kaminsky R, Schmid C, Lun ZR. Susceptibility of dyskinetoplastic Trypanosoma evansi and T. equiperdum to isometamidium chloride.. Parasitol Res 1997;83(8):816-8.
    doi: 10.1007/s004360050346pubmed: 9342750google scholar: lookup
  27. Lun ZR, Lai DH, Wen YZ, Zheng LL, Shen JL, Yang TB, Zhou WL, Qu LH, Hide G, Ayala FJ. Cancer in the parasitic protozoans Trypanosoma brucei and Toxoplasma gondii.. Proc Natl Acad Sci U S A 2015 Jul 21;112(29):8835-42.
    pmc: PMC4517281pubmed: 26195778doi: 10.1073/pnas.1502599112google scholar: lookup

Citations

This article has been cited 16 times.
  1. Boushaki D, Wallis J, Van den Broeck F, Schnaufer A. Molecular Analysis of Trypanosome Infections in Algerian Camels.. Acta Parasitol 2022 Sep;67(3):1246-1253.
    doi: 10.1007/s11686-022-00577-7pubmed: 35657485google scholar: lookup
  2. Desquesnes M, Gonzatti M, Sazmand A, Thévenon S, Bossard G, Boulangé A, Gimonneau G, Truc P, Herder S, Ravel S, Sereno D, Jamonneau V, Jittapalapong S, Jacquiet P, Solano P, Berthier D. A review on the diagnosis of animal trypanosomoses.. Parasit Vectors 2022 Feb 19;15(1):64.
    doi: 10.1186/s13071-022-05190-1pubmed: 35183235google scholar: lookup
  3. Oldrieve G, Verney M, Jaron KS, Hébert L, Matthews KR. Monomorphic Trypanozoon: towards reconciling phylogeny and pathologies.. Microb Genom 2021 Aug;7(8).
    doi: 10.1099/mgen.0.000632pubmed: 34397347google scholar: lookup
  4. Tanaka Y, Suganuma K, Watanabe K, Kobayashi Y. Pathology of female mice experimentally infected with an in vitro cultured strain of Trypanosoma equiperdum.. J Vet Med Sci 2021 Aug 6;83(8):1212-1218.
    doi: 10.1292/jvms.21-0056pubmed: 34135196google scholar: lookup
  5. Murata T, Batkhuu J. Biological activity evaluations of chemical constituents derived from Mongolian medicinal forage plants and their applications in combating infectious diseases and addressing health problems in humans and livestock.. J Nat Med 2021 Sep;75(4):729-740.
    doi: 10.1007/s11418-021-01529-7pubmed: 34018093google scholar: lookup
  6. Di Febo T, Krasteva I, Bonfini B, Tittarelli M, Matteucci O, Orsini G, Rossi E, Podaliri Vulpiani M, Rodomonti D, Iannetti L, Luciani M. Humoral immune response and delayed-type hypersensitivity in rabbits infected with Trypanosoma equiperdum.. Sci Rep 2020 Sep 10;10(1):14914.
    doi: 10.1038/s41598-020-71992-xpubmed: 32913248google scholar: lookup
  7. Tanaka Y, Adilbish A, Koyama K, Bayasgalan MO, Horiuchi N, Uranbileg N, Watanabe K, Purevdorj B, Gurdorj S, Banzragch B, Badgar B, Suganuma K, Yokoyama N, Inoue N, Kobayashi Y. Immunohistochemical phenotyping of macrophages and T lymphocytes infiltrating in peripheral nerve lesions of dourine-affected horses.. J Vet Med Sci 2020 Oct 20;82(10):1502-1505.
    doi: 10.1292/jvms.20-0172pubmed: 32788501google scholar: lookup
  8. Mizushima D, Amgalanbaatar T, Davaasuren B, Kayano M, Naransatsral S, Myagmarsuren P, Otgonsuren D, Enkhtaivan B, Davkharbayar B, Mungun-Ochir B, Baatarjargal P, Nyamdolgor U, Soyolmaa G, Altanchimeg A, Zoljargal M, Nguyen TT, Battsetseg B, Battur B, Inoue N, Yokoyama N, Suganuma K. Nationwide serological surveillance of non-tsetse-transmitted horse trypanosomoses in Mongolia.. Parasite Epidemiol Control 2020 Aug;10:e00158.
    doi: 10.1016/j.parepi.2020.e00158pubmed: 32642568google scholar: lookup
  9. Sereno D, Akhoundi M, Sayehmri K, Mirzaei A, Holzmuller P, Lejon V, Waleckx E. Noninvasive Biological Samples to Detect and Diagnose Infections due to Trypanosomatidae Parasites: A Systematic Review and Meta-Analysis.. Int J Mol Sci 2020 Feb 29;21(5).
    doi: 10.3390/ijms21051684pubmed: 32121441google scholar: lookup
  10. Akazue PI, Ebiloma GU, Ajibola O, Isaac C, Onyekwelu K, Ezeh CO, Eze AA. Sustainable Elimination (Zero Cases) of Sleeping Sickness: How Far Are We from Achieving This Goal?. Pathogens 2019 Aug 29;8(3).
    doi: 10.3390/pathogens8030135pubmed: 31470522google scholar: lookup
  11. Yasine A, Ashenafi H, Geldhof P, Van Brantegem L, Vercauteren G, Bekana M, Tola A, Van Soom A, Duchateau L, Goddeeris B, Govaere J. Histopathological lesions in reproductive organs, distal spinal cord and peripheral nerves of horses naturally infected with Trypanosoma equiperdum.. BMC Vet Res 2019 May 28;15(1):175.
    doi: 10.1186/s12917-019-1916-7pubmed: 31138270google scholar: lookup
  12. Büscher P, Gonzatti MI, Hébert L, Inoue N, Pascucci I, Schnaufer A, Suganuma K, Touratier L, Van Reet N. Equine trypanosomosis: enigmas and diagnostic challenges.. Parasit Vectors 2019 May 15;12(1):234.
    doi: 10.1186/s13071-019-3484-xpubmed: 31092285google scholar: lookup
  13. Davaasuren B, Yamagishi J, Mizushima D, Narantsatsral S, Otgonsuren D, Myagmarsuren P, Battsetseg B, Battur B, Inoue N, Suganuma K. Draft Genome Sequence of Trypanosoma equiperdum Strain IVM-t1.. Microbiol Resour Announc 2019 Feb;8(9).
    doi: 10.1128/MRA.01119-18pubmed: 30834384google scholar: lookup
  14. Mizushima D, Amgalanbaatar T, Davaasuren B, Molefe NI, Battur B, Battsetseg B, Inoue N, Yokoyama N, Suganuma K. The utility of an rTeGM6-4r-based immunochromatographic test for the serological diagnosis of non-tsetse-transmitted equine trypanosomosis in rural areas of Mongolia.. Parasitol Res 2018 Sep;117(9):2913-2919.
    doi: 10.1007/s00436-018-5982-8pubmed: 29943319google scholar: lookup
  15. Gizaw Y, Megersa M, Fayera T. Dourine: a neglected disease of equids.. Trop Anim Health Prod 2017 Jun;49(5):887-897.
    doi: 10.1007/s11250-017-1280-1pubmed: 28439783google scholar: lookup
  16. Suganuma K, Yamasaki S, Molefe NI, Musinguzi PS, Davaasuren B, Mossaad E, Narantsatsral S, Battur B, Battsetseg B, Inoue N. The establishment of in vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum.. Int J Parasitol Drugs Drug Resist 2017 Aug;7(2):200-205.
    doi: 10.1016/j.ijpddr.2017.04.002pubmed: 28437733google scholar: lookup
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.