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Journal of parasitic diseases : official organ of the Indian Society for Parasitology2024; 49(1); 121-129; doi: 10.1007/s12639-024-01744-5

Isolation of Toxoplasma gondii from the masseter muscles of equines destined for human consumption in a slaughterhouse in southern Brazil.

Abstract: The aim of this study was to isolate from equids destined for slaughter in a Brazilian slaughterhouse. A total of 354 equids were analyzed, with blood samples collected from all the animals and samples of masseter muscle and brain tissue collected from 319 animals. A serological test was conducted to detect equids with specific antibodies for . Molecular detection of by nested PCR was performed on the tissue samples collected. Tissue samples were tested by murine bioassay in an attempt to isolate either the parasite or the parasite DNA. Real-time PCR was performed on the brain samples from 11 mice which seroconverted after inoculation, to quantify the parasitic DNA. Genotyping was performed in masseter tissues samples from positive horses or from tissues of mice inoculated with masseter. The seroprevalence of infection was 19.2%. Nested PCR showed that 5.3% of the equines and 28.8% of the mice from the bioassays were positive for . Nine masseter muscle samples were positive (either on nested-PCR or in bioassay). Genotyping by PCR-RFLP was attempeted on all 12 isolates and was successful in seven, revealing six recombinant and one atypical genotype. The detection of DNA in masseter muscle tissue of horses destined for consumption clearly indicates a risk to human health.
© Indian Society for Parasitology 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
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Publication Date: 2024-09-25 PubMed ID: 39975622PubMed Central: PMC11832872DOI: 10.1007/s12639-024-01744-5Google Scholar: Lookup
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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 aimed to identify the presence of the parasite Toxoplasma gondii in the muscles of horses intended for human consumption in a Brazilian slaughterhouse.

Research Methods and Procedures

  • To achieve their objective, the researchers conducted tests on 354 horses that were headed for a slaughterhouse. They collected blood from each of these animals, while also obtaining samples of masseter muscle and brain tissue from 319 of them.
  • With these body fluid and tissue samples, the team performed serological tests to identify horses with specific antibodies for Toxoplasma gondii. This allowed them to determine whether the horses had been exposed to the parasite.
  • Molecular detection of Toxoplasma gondii was done via nested PCR (polymerase chain reaction) on the tissue samples. This is a highly accurate and sensitive technique used for amplifying and detecting specific DNA sequences.
  • Tissue samples were also tested using a murine bioassay, a method where samples are injected into mice to observe for any disease symptoms or seroconversion (development of specific antibodies to the injected antigen).
  • The team performed real-time PCR on brain samples from 11 mice that experienced seroconversion. This was done to quantify the amount of the parasite’s genetic material present.
  • The last stage involved genotyping of the masseter tissues from positive horses or the tissues from inoculated mice. Genotyping was performed using PCR-RFLP (polymerase chain reaction – restriction fragment length polymorphism), a technique that differentiates organisms based on the patterns their DNA fragments produce.

Findings

  • The results showed that 19.2% of the equines were seropositive for Toxoplasma gondii infection, indicating that they had been exposed to the parasite.
  • Through nested PCR, it was found out that 5.3% of the horses and a whopping 28.8% of the mice from the bioassays were positive for Toxoplasma gondii.
  • Of all the tests done, nine masseter muscle samples showed positive results, either via nested-PCR or through the bioassay.
  • When genotyping was attempted on all these 12 samples, it was successful in seven cases. Of these, six samples were recombinant and one displayed an atypical genotype.

Final Conclusion

  • This study successfully identified the presence of Toxoplasma gondii in masseter muscle tissue of horses meant for consumption, thereby suggesting a health risk for humans consuming the infected meat. Thus, the study recommends better screening procedures in slaughterhouses to alleviate such risks.

Cite This Article

APA
de Oliveira UV, Varjão JL, de Jesus Deiró AG, Maciel BM, Silva FL, Pinheiro AM, Gondim LFP, Munhoz AD. (2024). Isolation of Toxoplasma gondii from the masseter muscles of equines destined for human consumption in a slaughterhouse in southern Brazil. J Parasit Dis, 49(1), 121-129. https://doi.org/10.1007/s12639-024-01744-5

Publication

Journal of parasitic diseases : official organ of the Indian Society for Parasitology
ISSN: 0971-7196
NlmUniqueID: 9713059
Country: India
Language: English
Volume: 49
Issue: 1
Pages: 121-129

Researcher Affiliations

de Oliveira, Uillians Volkart
  • Faculdade de Ciências Sociais aplicadas, Itamaraju, Bahia 45,836-000 Brazil.
Varjão, José Luís
  • Departamento de Ciências Agrárias e Ambientais, Universidade Estadual de Santa Cruz, Ilhéus, Bahia Brazil.
de Jesus Deiró, Ana Graziela
  • Departamento de Ciências Agrárias e Ambientais, Universidade Estadual de Santa Cruz, Ilhéus, Bahia Brazil.
Maciel, Bianca Mendes
  • Departamento de Ciências Biológicas, Universidade Estadual de Santa Cruz, Ilhéus, Bahia Brazil.
Silva, Fabiana Lessa
  • Departamento de Ciências Agrárias e Ambientais, Universidade Estadual de Santa Cruz, Ilhéus, Bahia Brazil.
Pinheiro, Alexandre Moraes
  • Centro de Ciências Agrárias, Biológicas e Ambientais, Universidade Federal do Recôncavo Baiano, Cruz das Almas, Bahia Brazil.
Gondim, Luis Fernando Pita
  • Departamento de Anatomia, Patologia e Clínicas, Universidade Federal da Bahia, Salvador, Bahia Brazil.
Munhoz, Alexandre Dias
  • Departamento de Ciências Agrárias e Ambientais, Universidade Estadual de Santa Cruz, Ilhéus, Bahia Brazil.

Conflict of Interest Statement

Conflict of interestThe authors have no relevant financial or non-financial interests to disclose.

References

This article includes 48 references
  1. Al-khalidi NW, Dubey JP. Prevalence of Toxoplasma gondii Infection in Horses. J Parasitol 65(331):334.
    pubmed: 448624doi: 10.2307/3280181google scholar: lookup
  2. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402.
    pmc: PMC146917pubmed: 9254694doi: 10.1093/nar/25.17.3389google scholar: lookup
  3. Alvarado-esquivel C, Alvarado-esquivel D, Dubey JP. Prevalence of Toxoplasma gondii antibodies in domestic donkeys (Equus asinus) in Durango, Mexico slaughtered for human consumption. BMC Vet Res 11:6.
    pmc: PMC4301930pubmed: 25595816doi: 10.1186/s12917-015-0325-9google scholar: lookup
  4. Aroussi A, Vignoles P, Dalmay F, Wimel L, Dardé ML, Mercier A, Ajzenberg D. Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests. Parasite 22:14.
    pmc: PMC4374124pubmed: 25809058doi: 10.1051/parasite/2015014google scholar: lookup
  5. Ayres M, Junior MA, Ayres DL, Santos AAS. BioEstat: aplicações estatísticas nas áreas das ciências biomédicas Versão 4.0 Belém: IDSM. .
  6. Birkenheuer AJ, Levy MG, Andbreitschwerdt EB. Development and evaluation of a seminested PCR for detection and differentiation of Babesia gibsoni (Asian genotype) and B. canis DNA in canine blood samples. J Clin Microbiol 41:4172–7.
    pmc: PMC193857pubmed: 12958243
  7. Costa-Silva TA, Pereira-Chioccola VL. Fase aguda da infecção por Toxoplasma gondii: avaliação do parasitismo sanguíneo e resposta humoral em camundongos isogênicos. Sci Med 20(1):88–92.
  8. Dubey JP, Miller NL, Frenkel JK. The Toxoplasma gondii oocyst from cat feces. J Exp Med 132:636–662.
    pmc: PMC2138867pubmed: 4927658doi: 10.1084/jem.132.4.636google scholar: lookup
  9. Dubey JP. Long-term persistence of Toxoplasma gondii in tissues of pigs inoculated with Toxoplasma gondii oocysts and effect of freezing on viability of tissue cysts in pork. Am J Vet Res 49:910–913.
    pubmed: 3400928
  10. Dubey JP. Tissue cyst tropism in Toxoplasma gondii: a comparison of tissue cyst formation in organs of cats, and rodents fed oocysts. Parasitology 115:15–20.
    pubmed: 9226953doi: 10.1017/s0031182097008949google scholar: lookup
  11. Dubey JP. Advances in the life cycle of Toxoplasma gondii. Int J Parasitol 28:1019–1024.
    pubmed: 9724872doi: 10.1016/s0020-7519(98)00023-xgoogle scholar: lookup
  12. Dubey JP. Refinement of pepsin digestion method for isolation of Toxoplasma gondii from infected tissues. Vet Parasitol 74:75–77.
    pubmed: 9493311doi: 10.1016/s0304-4017(97)00135-0google scholar: lookup
  13. Dubey JP, Thulliez P, Romand S, Kwok OCH, Shen KS, Gamble HR. Serologic prevalence of Toxoplasma gondii in horses slaughtered for food in North America. Vet Parasitol 86:235–238.
    pubmed: 10536980doi: 10.1016/s0304-4017(99)00148-xgoogle scholar: lookup
  14. Dubey JP, Patitucci AN, Su C, Sundar N, Kwok OCH, Shen SK. Characterization of toxoplasma gondii isolates in free-range chickens from Chile, South America. Vet Parasitol 140:76–82.
    pubmed: 16672177doi: 10.1016/j.vetpar.2006.03.023google scholar: lookup
  15. Dubey JP, Jones JL. Toxoplasma gondii infection in humans and animals in the United States. Int J Parasitol 38:1257–1278.
    pubmed: 18508057doi: 10.1016/j.ijpara.2008.03.007google scholar: lookup
  16. Dubey JP. Toxoplasmosis of animals and humans, 2nd edn. CRC Press, Boca Raton, p 313. .
  17. Andreoletti O, Budka H, Buncic S, Colin P, Collins JD, De A, Noeckler BN, Maradona MP, Roberts T, Vågsholm I, Vanopdenbosch E. Surveillance and monitoring of Toxoplasma in humans, food and animals-scientific opinion of the panel on biological hazards. EFSA 583:1–64.
    doi: 10.2903/j.efsa.2007.583google scholar: lookup
  18. Elbez-Rubinstein A, Ajzenberg D, Dardé ML, Cohen R, Dumètre A, Yera H, Gondon E, Janaud JC, Thulliez P. Congenital toxoplasmosis and reinfection during pregnancy: case report, strain characterization, experimental model of reinfection, and review. J Infect Dis 199:280–285.
    pubmed: 19032062doi: 10.1086/595793google scholar: lookup
  19. Evers F, Garcia JL, Navarro IT, Zulpo DL, Nino BSL, Ewald MPC, Pagliari S, Almeida JC, Freire RL. Diagnosis and isolation of Toxoplasma gondii in horses from Brazilian slaughterhouses. Rev Bras Parasitol Vet 22:58–63.
    pubmed: 23538498doi: 10.1590/s198429612013005000009google scholar: lookup
  20. Gennari SM, Esmerini PO, Lopes MG, Soares HS, Vitaliano SN, Cabral AD, Villalobos EM. Occurrences of antibodies to Toxoplasma gondii and its isolation and genotyping in donkeys, mules and horses in Brazil. Vet Parasitol 209:129–132.
    pubmed: 25747488doi: 10.1016/j.vetpar.2015.01.023google scholar: lookup
  21. Habibi GR, Imani AR, Gholami MR, Hablolvarid MH, Behroozikhah AM, Lotfi M, Kamalzade M, Najjar E, Esmaeil-nia K, Bozorgi S. Detection and identification of Toxoplasma gondii type one infection in sheep aborted fetuses in Qazvin Province of Iran. Iran J Parasitol 7:64–72.
    pmc: PMC3469174pubmed: 23109964
  22. Hall T. BioEdit: an important software for molecular biology. GERF Bull Bios 2:60–61.
  23. Homan WL, Vercammen M, DeBraekeleer J, Verschueren H. Identification of a 200 to 300 fold repetitive 529 bp DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR. Int J Parasitol 30:69–75.
    pubmed: 10675747doi: 10.1016/s0020-7519(99)00170-8google scholar: lookup
  24. James KE, Smith WA, Packham AE, Conrad PA, Pusterla N. Toxoplasma gondii seroprevalence and association with equine protozoal myeloencephalitis: a case– control study. Vet J 224:38–43.
    pubmed: 28697873doi: 10.1016/j.tvjl.2017.05.008google scholar: lookup
  25. Kalambhe D, Gill J, Singh BB. Molecular detection of Toxoplasma gondii in the slaughter sheep and goats from North India. Vet Parasitol 241:35–38.
    pubmed: 28579027doi: 10.1016/j.vetpar.2017.05.009google scholar: lookup
  26. Klun I, Uzelac A, Villena I, Mercier A, Bobić B, Nikolić A, Rajnpreht I, Opsteegh M, Aubert D, Blaga R, Van der giessen J, Djurković-Djaković O. The first isolation and molecular characterization of Toxoplasma gondii from horses in Serbia. Parasit Vectors 10:167.
    pmc: PMC5379513pubmed: 28376902doi: 10.1186/s13071-017-2104-xgoogle scholar: lookup
  27. Lacerda LC, Silva AN, Freitas JS, Cruz RDS, Said RA, Munhoz AD. Feline immunodeficiency virus and feline leukemia virus: frequency and associated factors in cats in northeastern brazil. Genet Mol Res 16:1–8.
    pubmed: 28510253doi: 10.4238/gmr16029633google scholar: lookup
  28. Lélu M, Villena I, Dardé ML, Aubert D, Geers R, Dupuis E, Marnef F, Poulle ML, Gotteland C, Dumètre A, Gilot-Fromont E. Quantitative estimation of the viability of Toxoplasma gondii oocysts in soil. J Appl Environ Microbiol 78:5127–5132.
    pmc: PMC3416395pubmed: 22582074doi: 10.1128/aem.00246-12google scholar: lookup
  29. Lindsay DS, Blagburn BL, Braund KG. A review of Toxoplasma gondii and muscular toxoplasmosis. BAM 5:255–260.
  30. . Quantidade de abate anual por ano/espécie. .
  31. Martuzzi F, Catalano AL, Sussi C. Characteristics of horsemeat consumption and production in Italy. Ann Fac Med Vet, Univ Parma 21:213–233.
  32. Opsteegh M, Maas M, Schares G, Van der giessen J. Relationship between seroprevalence in the main livestock species and presence of Toxoplasma gondii in meat (GP/EFSA/BIOHAZ/ 2013/01). An extensive literature review. Final report. EFSA Supporting Publications 13.
  33. Papini RA, Buzzone G, Nardoni S, Rocchigiani G, Mancianti F. Seroprevalence and genotyping of Toxoplasma gondii in horses slaughtered for human consumption in Italy. J Equi Vet Sci 35:657–661.
    doi: 10.1016/j.jevs.2015.06.012google scholar: lookup
  34. Paştiu AI, Györke A, Kalmár Z, Bolfă P, Rosenthal BM, Oltean M, Villena I, Spînu M, Cozma V. Toxoplasma gondii in horse meat intended for human consumption in Romania. Vet Parasitol 212:393–395.
    pubmed: 26238654doi: 10.1016/j.vetpar.2015.07.024google scholar: lookup
  35. Pena HF, Gennari SM, Dubey JP, Su C. Population structure and mousevirulence of Toxoplasma gondii in Brazil. Int J Parasitol 38:561–569.
    pubmed: 17963770doi: 10.1016/j.ijpara.2007.09.004google scholar: lookup
  36. Pena HFJ, Pinheiro TM, Soares HS, Oliveira S, Alves BF, Ferreira MN, Gennari SM. Typical Brazilian genotype of Toxoplasma gondii isolated from a horse destined for human consumption in Europe from a slaughterhouse. Parasitol Res 117:3305–3308.
    pubmed: 29978419doi: 10.1007/s00436-018-5999-zgoogle scholar: lookup
  37. Pomares C, Ajzenberg D, Bornard L, Bernardin G, Hasseine L, Dardé L, Marty P. Toxoplasmosis and horse meat, France [letter]. Emerg Infect Dis 17:1327–1328.
    pmc: PMC3381409pubmed: 21762609doi: 10.3201/eid1707.101642google scholar: lookup
  38. Pereira PF, Barbosa AS, Santos ALC, Bolais PF, Dardé ML, Amendoeira MRR. Toxoplasma gondii: infection among shelter and stray cats in Rio de Janeiro. Brazil. Rev Bras Parasitol Vet 27:401–408.
    pubmed: 30208125doi: 10.1590/s1984-296120180061google scholar: lookup
  39. Portella LP, Cadore GC, Sangioni LA, Pellegrini LFV, Fighera R, Ramos F, Vogel FSF. Antibodies against Apixomplexa protozoa and absence sarcocysts in heart tissues from horses in southern Brazil. Rev Bras Parasitol Vet 26:100–103.
    pubmed: 28327879doi: 10.1590/s1984-29612016068google scholar: lookup
  40. Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci 74:5463–5467.
    pmc: PMC431765pubmed: 271968doi: 10.1073/pnas.74.12.5463google scholar: lookup
  41. Shaapan RM, Ghazy AA. Isolation of Toxoplasma gondii from horse meat in Egypt. Pak J Biol Sci 10:174–7.
    pubmed: 19070010
  42. Shwab EK, Zhu X-Q, Majumdar D, Pena HFJ, Gennari SM, Dubey JP, Su C. Geographical patterns of Toxoplasma gondii genetic diversity revealed by multilocus PCR-RFLP genotyping. Parasitology 141:453–461.
    pubmed: 24477076doi: 10.1017/s0031182013001844google scholar: lookup
  43. Su C, Shwab EK, Zhou P, Zhu XQ, Dubey JP. Moving towards an integrated approach to molecular detection and identification of Toxoplasma gondii. Parasitology 137:1–11.
    pubmed: 19765337doi: 10.1017/s0031182009991065google scholar: lookup
  44. Stanciu S. Horse meat consumption—between scandal and reality. Procedia Econ Financ 23:697–703.
    doi: 10.1016/s2212-5671(15)00392-5google scholar: lookup
  45. Tenter AM, Heckeroth AR, Weiss LM. Toxoplasma gondii: from animals to humans. Int J Parasitol 30:1217–1258.
    pmc: PMC3109627pubmed: 11113252doi: 10.1016/s0020-7519(00)00124-7google scholar: lookup
  46. Vidotto O, Kano FS, Freire RL, Mitisuka R, Ogawa L, Bonesi G, Navarro IT, Franciscon FSG. Ocorrência de anticorpos anti-Toxoplasma gondii em equinos procedentes de 4 estados (SP, PR, MS, MT) abatidos em Apucarana, no Paraná. Semina Cienc Agrár 18:9–13.
  47. Xing H, Xu L, Song X, Li X, Yan R. Seroprevalence of Toxoplasma gondii and Trichinella spiralis in horses in Xinjiang, Northwestern China. J Equi Vet Sci 60:11–15.
    doi: 10.1016/j.jevs.2017.06.006google scholar: lookup
  48. Yang N, Mu MY, Yuan GM, Zhang GX, Li HK, He JB. Seroprevalence of Toxoplasma gondii in slaughtered horses and donkeys in Liaoning province, northeastern China. Parasit Vectors 6:1–4.
    pmc: PMC3659062pubmed: 23680297doi: 10.1186/1756-3305-6-140google scholar: lookup

Citations

This article has been cited 1 times.
  1. Vela LA, Bravo Ramos JL, Rojas SB, Sánchez Otero MG, Montes SS, Ortiz Carbajal LA, Exsome CP, Baena AB. Detection of Toxoplasma Gondii and Risk Factors in Retail Meat in Southern Mexico. Acta Parasitol 2026 Jan 26;71(1):23.
    doi: 10.1007/s11686-025-01214-9pubmed: 41586924google scholar: lookup
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