Detección de leptospiras patógenas en tejido renal de ratón casero (Mus musculus) en Sincelejo, Colombia

Abstract

Leptospirosis is a zoonotic disease of worldwide distribution with more frequency in tropical countries. Some rodents and among them the Mus musculus or house mouse are asymptomatic chronic carriers of pathogenic Leptospira spp because they have them in their kidneys and spread it to the environment through the urine. This study aimed to detect Leptospira spp molecularly in kidneys removed from M. musculus in the municipality of Sincelejo (Sucre, Colombia). The capture of rodents was conducted in the urban area of the city: intra and peridomiciliar Sherman® traps were installed in the second quarter of 2010 and the first quarter of 2011 for 14 to 15 hours. During the necropsy the kidneys of individuals were removed for DNA extraction and PCR analysis for the amplification of the 423 pb gene segment between positions 270 and 692 of the coding region LipL32. A total of 154 individuals were captured, of which five were positive for pathogenic Leptospira spp. The correlation analysis showed that mice are mostly positive in commune 2, followed by commune 4 in the municipality. The study suggests that basic sanitation establishes favorable environments for the epidemiological process of the bacteria and the circulation of pathogenic Leptospira spp. in M. musculus is demonstrated in the municipality of Sincelejo. It is inferred that rodent infestation in the studied area is a potential source for the transmission of pathogenic Leptospira spp. to human beings, which is susceptible to all pathogenic species of the genus.

Keywords

leptospirosis

Leptospira

Mus musculus

PCR

LipL32

References

1. Bharti AR, Nally JE, Ricaldi JN, Matthias MA, Diaz MM, Lovett MA, et al. Leptospirosis: a zoonotic disease of global importance. The Lancet infectious diseases. 2003;3(12):757-71.

2. Fornazari F, da Silva RC, Richini-Pereira VB, Beserra HEO, Luvizotto MCR, Langoni H. Comparisonof conventional PCR, quantitative PCR, bacteriological culture and the Warthin Starry technique to detect Leptospira spp. in kidney and liver samples from naturally infected sheep from Brazil. Journal of microbiological methods. 2012;90(3):321-6.

3. Sacsaquispe R, Glenny M, Céspedes M. Estudio preliminar de leptospirosis en roedores y canes en salitral, Piura-1999. Revista Peruana de Medicina Experimental y Salud Pública. 2003;20(1):39-40.

4. Lomar AV, Diament D, Torres JR. Leptospirosis in Latin America. Infectious disease clinics of North America. 2000;14(1):23-39.

5. Collares-Pereira M, Mathias M, Santos-Reis M, Ramalhinho M, Duarte-Rodrigues P. Rodents and Leptospira transmission risk in Terceira island (Azores). European journal of epidemiology. 2000;16(12):1151-7.

6. Agudelo-Flórez P, Arango JC, Merizalde E, Londoño AF, Quiroz VH, Rodas JD. Evidencia serológica de circulación de Leptospira spp en Rattus norvegicus naturalmente expuestos en una zona urbana colombiana. Revista de Salud Pública. 2010;12(6):990-9.

7. Agudelo-Flórez P, Londoño AF, Quiroz VH, Ángel JC, Moreno N, Loaiza ET, et al. Prevalence of Leptospira spp. in urban rodents from a groceries trade center of Medellin, Colombia. The American journal of tropical medicine and hygiene. 2009;81(5):906-10.

8. Ríos R, Franco S, Mattar S, Urrea M, Tique V. Seroprevalence of Leptospira sp., Rickettsia sp. and Ehrlichia sp. in rural workers of Sucre, Colombia. Infectio. 2008;12(2):90-5.

9. Ferro BE, Rodriguez AL, Perez M, Travi BL. [Seroprevalence of Leptospira infection in habitants of peripheral neighborhoods in Cali, Colombia]. Biomedica : revista del Instituto Nacional de Salud. 2006;26(2):250-7.

10. Romero Almaraz MdL, Sánchez Hernández C, García Estrada C, Owen RD. Mamíferos pequeños. Manual de técnicas de captura, preparación, preservación y estudio: Las Prensas de Ciencias. Coordinación de Servicios Editoriales, Facultad de Ciencias, UNAM; 2007.

11. Mills JN, Childs JE, Ksiazek TG, Peters C, Velleca WM. Methods for trapping and sampling small mammals for virologic testing: US Department of Health and Human Services, Public Health Service, Centres for Disease Control and Prevention; 1995.

12. Natarajaseenivasan K, Vijayachari P, Sharma S, Sugunan AP, Vedhagiri K, Selvin J, et al. FlaB PCRbased identification of pathogenic leptospiral isolates. Journal of Microbiology, mmunology and Infection. 2010;43(1):62-9.

13. Marder G, Ruiz R, LM-Zorzo L, Merino D. Detección de leptospiras en riñon de roedores de la ciudad de Corrientes: estudio preliminar.

14. Emmons L, Feer F. Neotropical rainforest mammals: a field guide. 1997.

15. Nowak RM. Walker’s Mammals of the World: JHU Press; 1999.

16. Watts P. Extraction of DNA from tissue: High salt method.Liverpool. School of Biological Sciences. 2001;Version 1.0: 5.

17. Lopera-Barrero NM, Povh JA, Ribeiro RP, Gomes PC, Jacometo CB, Silva Lopes Td. Comparación de protocolos de extracción de ADN con muestras de aleta y larva de peces: extracción modificada con cloruro de sodio. Ciencia e investigación agraria. 2008;35(1):77-86.

18. Levett PN, Morey RE, Galloway RL, Turner DE, Steigerwalt AG, Mayer LW. Detection of pathogenic leptospires by real-time quantitative PCR. Journal of Medical Microbiology. 2005;54(1):45-9.

19. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: Molecular Evolutionary Genetics Analysis Using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular biology and evolution. 2011;28(10):2731-9.

20. Di Rienzo JAC, F. Balzarini, M. G. Gonzalez, L. Tablada, M. Robledo, C. W. InfoStat. 2011.

21. Johnson MA, Smith H, Joseph P, Gilman RH, Bautista CT, Campos KJ, et al. Environmental exposure and leptospirosis, Peru. Emerging infectious diseases. 2004;10(6).

22. Donaires LF, Céspedes MJ, Sihuincha MG, Pachas PE. Determinantes ambientales y sociales para la reemergencia de la leptospirosis en la región amazónica del Perú, 2012. Revista Peruana de Medicina Experimental y Salud Pública. 2012;29(2):280-4.

23. Nardone A, Capek I, Baranton G, Campèse C, Postic D, Vaillant V, et al. Risk factors for leptospirosis in metropolitan France: results of a national case-control study, 1999–2000. Clinical Infectious Diseases. 2004;39(5):751-3.

24. Houemenou G, Ahmed A, Libois R, Hartskeerl R. Leptospira spp. prevalence in small mammal populations in Cotonou, Benin. ISRN Epidemiology. 2013;2013.

25. Moreno N, Agudelo-Flórez P. Aplicación de las pruebas de PCR convencional simple y múltiple para la identificación de aislamientos de Leptospira spp. en Colombia. Revista Peruana de Medicina Experimental y Salud Pública. 2010;27(4):548-56.

26. Guerreiro H, Croda J, Flannery B, Mazel M, Matsunaga J, Reis MG, et al. Leptospiral proteins recognized during the humoral immune response to leptospirosis in humans. Infection and immunity. 2001;69(8):4958-68.

27. Haake DA, Matsunaga J. Characterization of the leptospiral outer membrane and description of three novel leptospiral membrane proteins. Infection and immunity. 2002;70(9):4936-45.

28. Zhang X-Y, Yu Y, He P, Zhang Y-X, Hu B-Y, Yang Y, et al. Expression and comparative analysis of genes encoding outer membrane proteins LipL21, LipL32 and OmpL1 in epidemic leptospires. Acta biochimica et biophysica Sinica. 2005;37(10):649-56.

29. Jung LRC, Bomfim MRQ, Kroon EG, Nunes ÁC. Identification of Leptospira serovars by RFLP of the RNA polymerase beta subunit gene (rpoB). Brazilian Journal of Microbiology. 2015;46(2):465-76.

30. Romero-Vivas CM, Thiry D, Rodríguez V, Calderón A, Arrieta G, Máttar S, et al. Molecular serovar characterization of Leptospira isolates from animals and water in Colombia. Biomédica. 2013;33:179-84.