Isolation and identification of Yersinia enterocolitica in bovine milk samples of bulk tanks from dairy farms in São Paulo, Brazil

Amanda Bezerra Bertolini, Marcelo Augusto Orsi Dutra, Lívia Maísa Guiraldi, Wesley José dos Santos, Maria Izabel Merino de Medeiros, Simone Baldini Lucheis

Abstract


Milk and its derivatives are good substrates for the proliferation of pathogenic and quality-deteriorating microorganisms, demanding rigorous care with milking, processing, and storage. Among the various bacteria that can grow in raw refrigerated milk, Yersinia enterocolitica, is an invasive enteropathogen of humans. This bacterium can cause a number of intestinal and extraintestinal clinical symptoms, ranging from mild gastroenteritis to mesenteric lymphadenitis, similar to appendicitis. To evaluate the prevalence of pathogenic Yersinia enterocolitica in raw milk from bulk milk tanks located in the State of São Paulo, 102 bovine milk samples (one per dairy farm) were evaluated by microbiological analyses, followed by biochemical tests PCR and genetic sequencing. Microbiological testing did not isolate Y. enterocolitica. However, PCR analysis revealed six samples that were positive for Y. enterocolitica (5.9%), confirmed by genetic sequencing. Only the inv gene was detected, which is present in virulent and avirulent Y. enterocolitica strains. There was great difficulty in microbiological isolation due to the difficulty of competitiveness of Y. enterocolitica in a very rich microbiota of raw milk. Although virulence genes known to be present in potentially pathogenic strains of Y. enterocolitica have not been identified, the presence of this pathogen in milk from expansion tanks, identified through PCR and confirmed by genetic sequencing, suggests that Y. enterocolitica may be a risk to public health, especially if milk and its derivatives are consumed without heat treatment.

Keywords


Microbiological analyses; Molecular diagnosis; Bovine milk; Public health; Yersiniosis.

Full Text:

PDF

References


ARCURI, E. F.; SILVA, P. D. L.; BRITO, M. P. V. P.; BRITO, J. R. F.; LARNGE, C. C.; MAGALHÃES, M. M. A. Contagem, isolamento e caracterização de bactérias psicrotróficas contaminantes de leite cru refrigerado. Ciência Rural, Santa Maria, v. 38, n. 8, p. 2250-2255, 2008.

BALAKRISHNA, K.; RADHIKA, M.; MURALI, H. S. Specific identification of pathogenic Yersinia enterocolitica by monoclonal antibodies generated against recombinant attachment invasion locus (rAil) protein. World Journal of Microbiology and Biotechnology, Berlin, v. 28, n. 2, p. 533-539, 2012.

BIER, O. Bacteriologia e imunologia. 17. ed. São Paulo: Melhoramentos, 1984. 1968 p.

BORRALHO, E.; ALBUQUERQUE, M. C.; CARDOSO, M. Pesquisa de Yersinia enterocolitica em linguiças frescas de porco em Porto Alegre, RS. Ciência Rural, Santa Maria, v. 29, n.4, p. 727-729, 1999.

BRASIL. Ministério da Agricultura Pecuária e Abastecimento. Decreto nº 9. 013, de 29 de março de 2017. Regulamenta a Lei nº 1. 283, de 18 de dezembro de 1950, e a Lei nº 7. 889, de 23 de novembro de 1989, que dispõem sobre a inspeção industrial e sanitária de produtos de origem animal. Diário Oficial [da] União, Brasília, 2017. Seção 1, p. 3. Disponível em: http://www.agricultura.gov.br. Acesso em: 2 aug. 2017.

DALLAL, M. M. S.; TABARRAIE, A.; MOEZARDALAN, K. Comparison of four methods for isolation of Yersinia enterocolitica from raw and pasteurized milk from northern Iran. International Journal of Food Microbiology, Amsterdam, v. 94, n. 1, p. 87-91, 2004.

DARWISH, S. F.; ASFOUR, H. A. E.; ALLAM, H. A. Incidence of Yersinia enterocolitica and Yersinia pseudotuberculosis in raw milk samples of different animal species using conventional and molecular methods. Alexandria Journal of Veterinary Sciences, Alexandria, v. 44, n. 1 p. 174-185, 2015.

DEAN, A. G.; SULLIVAN, K. M.; SOE, M. M. OpenEpi: open source epidemiologic statistics for public health. Update 2015/05/04. (Programa Estatístico online). Available at: http://www.OpenEpi.com. Accessed at: 16 oct. 2015.

DRUMMOND, N.; MURPHY, B. P.; RINGWOOD, T.; PRENTICE, M. B.; BUCKLEY, J. F.; FANNING, S. Yersinia enterocolitica: a brief review of the issues relating to the zoonotic pathogen, public health challenges, and the pork production chain. Foodborne Pathogens and Disease, Larchmont, v. 9, n. 3, p. 174-189, 2012.

FALCÃO, J. P.; FALCÃO, D. P.; DIAS, A. M. G.; CORREA, E. F. Microbiological quality of ice used to refrigerate foods. Food Microbiology, London, v. 19, n. 4, p. 269-276, 2002.

FALCÃO, J. P.; FALCÃO, D. P.; SILVA, A. P.; MALASPINA, A. C.; BROCCHI, M. Molecular typing and virulence markers of Yersinia enterocolitica, strains, from human, animal and food, origins isolated between 1968 and 2000 in Brazil. Journal of Medical Microbiology, Edinburgh, v. 55, n. 11, p. 1539-1548, 2006.

HANIFIAN, S.; KHANI, S. Prevalence of virulent Yersinia enterocolitica in buck raw milk and retail cheese in northern-west of Iran. International Journal of Food Microbiology, Amsterdam, v. 155, n. 2, p. 89-92, 2012.

KAYLEGIAN, K. E.; MOAG, R.; GALTON, D. M.; BOOR, K. J. Raw milk consumption beliefs and practices among New York State dairy producers. Food Protection Trends, Des Moines, v. 2, n. 3, p. 184-191, 2008.

MARTINS, K. B.; MARTINS-FACCIOLI, P. Y.; RIBOLI, D. F. M.; PEREIRA, C. V.; FERNANDES, S.; OLIVEIRA, A. A.; DANTAS, A.; ZAFALON, L. F.; CUNHA, M. L. R. S. Clonal profile, virulence and resistance of Staphylococcus aureus isolated from sheep milk. Brazilian Journal of Microbiology, São Paulo, v. 46, n. 2, p. 533-543, 2015.

MATTOCK, E.; BLOCKER, A. J. How do the virulence factors of shigella work together to cause disease? Frontiers in cellular and infection microbiology, Bristol v. 64, n. 7, p. 1-24, 2017.

PADILHA, M. R. F.; FERNANDES, Z. F.; LEAL, A. T. C. A.; LEAL, N. C.; ALMEIDA, A. M. P. Pesquisa de bactérias patogênicas em leite pasteurizado tipo C comercializado na cidade do Recife, Pernambuco Brasil. Revista da Sociedade Brasileira de Medicina Tropical, Rio de Janeiro, v. 34, n. 2, p. 67-171, 2001.

PINTO, P. S. A.; TEODORO, V. A. M.; VANETTI, M. C. D.; BELIVALACQUA, P. D.; MORAES, M. P.; PINTO, M. S. Aplicação de técnica de PCR na detecção de Yersinia enterocolitica em suínos abatidos sem inspeção. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, Belo Horizonte, v. 58, n. 1, p. 9-14, 2006.

RASMUSSEN, H. N.; OLSEN, J. E.; RASMUSSEN, O. F. RAPD analysis of Yersinia enterocolitica. Letters in Applied Microbiology, Oxford, v. 19, n. 19, p. 359-362, 1994.

RUEGG, P. L. Practical food safety interventions for dairy production. Journal of Dairy Science, Lancaster, v. 86, p. E1-E9, 2003. Supplement Special.

RUSAK, L. A.; REIS, F. C. M. F.; BARBOSA, A. V.; SANTOS, A. F. M.; PAIXÃO, R.; HOLER, E.; VALLIM, D. C.; ASENSI, M. D. Phenotypic and genotypic analysis of bio-serotypes of Yersinia enterocolitica from various sources in Brazil. Journal of Infection in Developing Countries, Rio de Janeiro, v. 8, n. 12, p. 1533-1540, 2014.

L, M.; SALONEN, M.; PULKKINEN, H.; HUUSKONEN, M.; HELLSTROM, S.; REVEZ, J.; HANNINEN, M. L.; FREDRIKSSON-AHOMAA, M.; LINDSTROM, M. Pathogenic bacteria in finnish bulk tank milk. Foodborne Pathogens and Disease, Larchmont, v. 10, n. 2 p. 99-106, 2013.

SILVA JUNIOR, E. A. Manual de controle sanitário em serviços de alimentação. 7. ed. São Paulo: Varela, 2014.

SOLTAN-DALLAL, M. M.; TABARRAIE, A.; MOEZARDALAN, K. Comparison of four methods for isolation of Yersinia enterocolitica from raw and pasteurized milk from northern Iran. International Journal of Food Microbiology, Amsterdam, v. 94, n. 1, p. 87-91, 2004.

STENKOVA, A. M.; ISAEVA, M. P.; RASSKAZOV, V. A. Development of a multiplex PCR procedure for detection of Yersinia genus with identification of pathogenic species (Y. pestis, Y. pseudotuberculosis, and Y. enterocolitica). Molecular Genetics, Microbiology and Virology, New York, v. 23, n. 3, p. 119-125, 2008.

TAVARES, A. B.; SOUZA, A. I. A.; DULAC, C. F.; MOREIRA, L. M.; DOMINGUEZ, L.; GONZALEZ, H. L.; CERESER, N. D.; TIMM, C. D. Fontes de contaminação de Yersinia enterocolitica durante a produção de leite. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, Belo Horizonte, v. 69, n. 2, p. 483-490, 2017.

TEODORO, V. A. M.; PINTO, P. S. A.; VANETTI, M. C. D.; BEVILACQUA, M. P.; PINTO, M. S. Aplicação da técnica de PCR na detecção da Yersinia enterocolitica em suínos abatidos sem inspeção. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, Belo Horizonte, v. 58, n. 1, p. 9-14, 2006.

THOERNER, P.; BIN KINGOMBE, C. I.; BOGLI-STUBER, K.; BISSIG-CHOISAT, B.; WASSENAAR, T. M.; FREY, J.; JEMMI, T. PCR detection of virulence gene distribution. Applied and Environmental Microbiology, Washington, v. 69, n. 3, p. 1810-1816, 2003.

TRABULSI, L. R.; ALTERTHUM, F. Microbiologia. 6. ed. São Paulo: Atheneu, 2015.

VIDON, D.; DELMAS, C. L. Incidence of Yersinia enterocolitica in raw milk in Eastern France. Applied and Environmental Microbiology, Washington, v. 41, n. 2, p. 355-359, 1981.

WAUTERS, G.; KANDOLO, K.; JANSSENS, M. Revised biogrouping scheme of Yersinia enterocolitica. Contributions to Microbiology and Immunology, Basel, v. 9, n. 4, p. 14-21, 1987.




DOI: http://dx.doi.org/10.5433/1679-0359.2019v40n2p723

Semina: Ciênc. Agrár.
Londrina - PR
E-ISSN 1679-0359
DOI: 10.5433/1679-0359
E-mail: semina.agrarias@uel.br
Este obra está licenciado com uma Licença Creative Commons Atribuição-NãoComercial 4.0 Internacional