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Tropical Animal Health and Production

, Volume 50, Issue 4, pp 883–888 | Cite as

Detection of bovine carriers of Leptospira by serological, bacteriological, and molecular tools

  • Melissa H. Pinna
  • Gabriel Martins
  • Ana Paula Loureiro
  • Walter Lilenbaum
Regular Articles

Abstract

Bovine leptospirosis is an important infectious disease that causes reproductive problems and economic risks, particularly in the tropics. The present study aimed to determine the extent of Leptospira infection among bovines on a slaughterhouse from Rio de Janeiro, Brazil via serological, bacteriological, and molecular tests. Two hundred eight bovines were examined in total, and we obtained 208 blood samples for serology, 198 urine samples collected via direct bladder puncture for polymerase chain reaction (PCR) and culture, 208 kidney samples (one from each animal) for PCR and culture, and 92 vaginal fluid samples from sterile swabs for PCR and culture. Serology demonstrated that 77/208 (37%) of the animals presented anti-Leptospira antibodies. Serogroup Sejroe was by far the most common. One hundrd thirty-three animals (63.9%) were PCR positive in at least one of the tested samples and were considered as Leptospira carriers. Furthermore, ten isolates were obtained by pure culture, all of them from urine samples. Bovine leptospirosis is widely prevalent, and the occurrence of renal carriers was unexpectedly much higher than generally reported.

Keywords

Leptospirosis Diagnostic Slaughtered PCR Isolation 

Notes

Acknowledgements

The authors are thankful to Camila Hamond, Professor Bruno Penna (UFF), and Sabrina Thomé for their assistance. This study was supported by Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). Walter Lilenbaum is CNPq fellows.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Abdollahpour, G., English, A. W. and Tasler, J. 1996. Isolation of Leptospira interrogans serovar grippotyphosa from a heifer in New South Wales. Australian Veterinary Journal, 73, 109–110CrossRefPubMedGoogle Scholar
  2. Adler B and De La Peña Moctezuma A. 2010. Leptospira and leptospirosis. Veterinary Microbiology, 140, 287–296CrossRefPubMedGoogle Scholar
  3. Azizi S., Tajbakhsh E., Hajimirzaei M.R., Varnamkhasti M.G., Sadeghian H. and Oryan A. 2012. Evaluation of ‘white-spotted kidneys’ associated with leptospirosis by polymerase chain reaction based LipL32 gene in slaughtered cows. Journal of the South African Veterinary Association, 83 Suppl 1, 1–5Google Scholar
  4. Balamurugan V., Gangadhar N.L., Mohandoss N., Thirumalesh S.R.A., Dhar M., Shome R. and Rahman H. 2013. Characterization of leptospira isolates from animals and humans: phylogenetic analysis identifies the prevalence of intermediate species in India. Springer Plus, 2, 362CrossRefPubMedPubMedCentralGoogle Scholar
  5. Bomfim M.R.Q., Barbosa-Stancioli E.F. and Koury M.C. 2008. Detection of pathogenic leptospires in urine from naturally infected cattle by nested PCR. The Veterinary Journal, 178, 251–256CrossRefPubMedGoogle Scholar
  6. Chiarelli D.C., Cosate M.R.V., Moreira E.C., Leite R.C., Lobato F.C.F., Silva J.A., Teixeira J.F.B. and Marcelino A.P. 2012. Controle da leptospirose em bovinos de leite com vacina autógena em Santo Antônio do Monte, Minas Gerais [Control of leptospirosis in dairy cattle with autogenous vaccine in Santo Antônio do Monte, MG, Brazil]. Pesquisa Veterinária Brasileira, 32 Supp l7, 633–639CrossRefGoogle Scholar
  7. Corney, B. G., Slack, A. T., Symonds, M. L., Dohnt, M. F., McClintock, C. S., McGowan, M. R., & Smythe, L. D. (2008). Leptospira weilii serovar Topaz, a new member of the Tarassovi serogroup isolated from a bovine source in Queensland, Australia. International Journal Of Systematic And Evolutionary Microbiology, 58 Suppl. 10, 2249–2252.CrossRefPubMedGoogle Scholar
  8. Dhaliwal G.S., Murray R.D. and Dobson H. 1996. Presence of antigen and antibodies in serum and genital discharges of heifers after experimental intrauterine inoculation with Leptospira interrogans serovar Hardjo. Research in Veterinary Science, 60, 157–162CrossRefPubMedGoogle Scholar
  9. Director A, Penna B, Hamond C, Loureiro A.P, Martins G, Medeiros M.A. and Lilenbaum W. 2014. Isolation of Leptospira interrogans Hardjoprajitno from vaginal fluid of a clinically healthy ewe suggests potential for venereal transmission. Journal of Medical Microbiology, 63, 1234–1236CrossRefPubMedGoogle Scholar
  10. Ellis W.A. 2015. Animal leptospirosis, p. 99–137. In: Adler B. (Ed.), Current Topics in Microbiology and Immunology. Vol. 387. (Springer, Berlin, Heidelberg).Google Scholar
  11. Ellis W.A., McParland P.J., Bryson D.G., Thiermann A.B. and Montgomery J. 1986. Isolation of leptospires from the genital tract and kidneys of aborted sows. Veterinary Record, 118, 294–295CrossRefPubMedGoogle Scholar
  12. Faine S., Adler B., Bolin C. & Perolat P. 2000. Leptospira and Leptospirosis. 2nd ed. (Medical Sciences, Melbourne. 296 p).Google Scholar
  13. Fornazari F., Silva R.C., Richini-Pereira V.B., Beserra H.E.O., Luvizotto M.C. and Langoni H. 2012. Comparison of conventional PCR, quantitative PCR, bacteriological culture and the Warthin Starry technique to detect Leptospira spp. in kidney and liver sample from naturally infected sheep from Brazilian Journal of Microbiology, 90, 321–26Google Scholar
  14. Gamage C.D., Koizumi N., Muto M., Nwafor-Okoli C., Kurukurusuriya S., Rajapakse J.R.P.V., Kularatne S.A.M., Kanda K., Lee R.B., Obayashi Y., Watanabe H. and Tamashiro H. 2011. Prevalence and carrier status of leptospirosis in smallholder dairy cattle and peridomestic rodents in Kandy, Sri Lanka. Vector-Borne and Zoonotic Diseases, 11 Suppl 8, 1041–147CrossRefPubMedGoogle Scholar
  15. Gamage C.D., Koizumi N., Perera A.K.C., Muto M., Nwafor-Okoli C., Ranasinghe S., Kularatne S.A.M., Rajapakse R.P.V.J., Kanda K., Lee R.B., Obayashi Y., Ohnishi M. and Tamashiro H. 2014. Carrier Status of Leptospirosis Among Cattle in Sri Lanka: A Zoonotic Threat to Public Health. Transboundary and Emerging Diseases, 61:91–96CrossRefPubMedGoogle Scholar
  16. Genovez M.E., Escócio C., Castro V., Gabriel F.H.L., Chiebao D.P. and Azevedo S.S. 2011. Fatores de risco associados à infecção pela Leptospira spp. Sorovar Hardjo em rebanhos exclusivos de ovinos e nos consorciados com bovinos. (Risk factors associated to Leptospira spp. serovar Hardjo infection in sheep-only herds and in sheep-cattle herds). Arquivos Instituto Biológico, São Paulo, 78 Suppl 4, 587–592Google Scholar
  17. Hamond C, Martins G., Loureiro A.P., Pestana C., Lawson-Ferreira R., Medeiros M.A. and Lilenbaum W. 2014. Urinary PCR as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock. Veterinary Research Communications, 38, 81–85CrossRefPubMedGoogle Scholar
  18. Hamond C., Pestana C.P., Medeiros M.A and Lilenbaum, W. 2015a. Genotyping of Leptospira directly in urine samples of cattle demonstrates a diversity of species and strains in Brazil. Epidemiology & Infection, 144, 1–4Google Scholar
  19. Hamond C., Martins G., Lilenbaum W., Pinna M. and Medeiros M.A. 2015b. Infection by Leptospira spp. in Cattle in a Tropical Region, Rio de Janeiro. The American Journal of Tropical Medicine and Hygiene, 92, 210–210CrossRefPubMedPubMedCentralGoogle Scholar
  20. Hernández-Rodríguez P., Díaz C. A., Dalmau E. A. and Quintero G. M. 2011. A comparison between polymerase chain reaction (PCR) and traditional techniques for the diagnosis of leptospirosis in bovines. Journal of Microbiological Methods, 8, 1–7CrossRefGoogle Scholar
  21. IBGE Sistema IBGE de Recuperação Automática. Instituto Brasileiro de Geografia e Estatística. Available in 2012. <www.sidra.ibge.gov.br> of subordinate document. Accessed 04 Abr 2017
  22. Levett P.N. 2015. Systematics of Leptospiraceae, p 11–20. In: Adler B. (Ed.), Current Topics in Microbiology and Immunology. Vol. 387. Springer, Berlin HeidelbergGoogle Scholar
  23. Lilenbaum W. and Santos M.R.C. 1996. Effect of management systems on the prevalence of bovine leptospirosis. Veterinary Record, 138, 570–571CrossRefPubMedGoogle Scholar
  24. Lilenbaum W., Varges R., Medeiros L., Cordeiro A.G., Cavalcanti A., Souza G.N., Richtzenhain L. and Vasconcellos S.A. 2008. Risk factors associated with leptospirosis in dairy goats under tropical conditions in Brazil. Research in Veterinary Science, 84, 14–17CrossRefPubMedGoogle Scholar
  25. Martins G. and Lilenbaum W. 2013. The panorama of animal leptospirosis in Rio de Janeiro, Brazil, regarding the seroepidemiology of the infection in tropical regions. BMC Veterinary Research, 9, 237CrossRefPubMedPubMedCentralGoogle Scholar
  26. Martins G., Loureiro A. P., Hamond C., Pinna M.H., Bremont S., Bourhy P. and Lilenbaum W. 2014. First isolation of Leptospira noguchii serogroups Panama and Autumnalis from cattle. Epidemiol Infection, 143 Suppl 7, 1–4Google Scholar
  27. Mineiro A.L.BB., Vieira R.J., Costa E.A., Santos R.L, Gonçalves L.M.F., Carvalho S.M., Bomfim M.R.Q. and Costa F.A.L. 2011. Serology, polymerase reaction and histopathology for leptospirosis in sample collected at slaughter dairy cows of Parnaiba region, state of Piauí, Brazil. Pesquisa Veterinária Brasileira, 31 Suppl 10, 859–866CrossRefGoogle Scholar
  28. Mughini-Gras L., Bonfanti L., Natale A., Comin A., Ferronato A., La Greca E., Patregnani T., Lucchese L. and Marangon S. 2014. Application of an integrated outbreak management plan for the control of leptospirosis in dairy cattle herds. Epidemiology & Infection, 142 Suppl 6, 1172–1181CrossRefGoogle Scholar
  29. OIE. Manual of diagnostic tests and vacines for terrestrial animals. World Organization for Animal Health, Paris, 2014. http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.01.12_LEPTO.pdf. of subordinate document. Accessed 28 Abr 2017
  30. Oliveira F.C.S., Azevedo S.S., Pinheiro S.R., Batista C.S.A., Moraes Z.M., Souza G.O., Gonçalves A.P. and Vasconcellos S.A. 2010. Fatores de risco para a leptospirose em fêmeas bovinas em idade reprodutiva no Estado da Bahia, nordeste do Brasil [Risk factors associated with leptospirosis were identified in cows in the state of Bahia, northeastern Brazil]. Pesquisa Veterinária Brasileira, 30 Suppl 5, 398–402CrossRefGoogle Scholar
  31. Oliveira F S., Oliveira D., Martins Filho E F, Costa F, Ristow P, Pinna M H, Lilenbaum W. 2016. Avaliação histológica e imuno-histoquímica da colonização vaginal por Leptospira em vacas com fluido vaginal positivo à PCR. Revista Brasileira de Medicina Veterinária, 38(Supl.1):163–167.Google Scholar
  32. Otaka D., Penna B., Martins G. and Lilenbaum W. 2012. Serology and PCR for bovine leptospirosis: a herd and individual approaches. Veterinary Record, 170, 338CrossRefPubMedGoogle Scholar
  33. Picardeau, M. 2013. Diagnosis and epidemiology of leptospirosis. Médecine et maladies infectieuses, 43, 1–9CrossRefPubMedGoogle Scholar
  34. Picardeau M., Bertherat E., Jancloes M., Skouloudis A.N., Durski K. and Hartskeerl R.A. 2014. Rapid tests for diagnosis of leptospirosis: current tools and emerging technologies. Diagnostic Microbiology and Infectious Disease, 78 Suppl 1 1–8CrossRefPubMedGoogle Scholar
  35. Ryan E.G., Leonard N., O’grady L., More S.J. and Doherty M.L. 2012. Seroprevalence of Leptospira Hardjo in the Irish suckler cattle population. Irish Veterinary Journal, 65 Suppl 8, 1–11Google Scholar
  36. Silva É.F., Cerqueira G.M., Seyffert N., Seixas F.K., Hartwig D.D., Ahanazio D.A., Pinto L.S., Queiroz A., Ko A.I., Brod C.S., and Dellagostin O.A. 2009. Leptospira noguchii and human and animal leptospirosis, Southern Brazil. Emerging Infectious Diseases, 15: 621–623.CrossRefPubMedPubMedCentralGoogle Scholar
  37. Stoddard R.A., Gee J.E., Wilkins P.P., Mccaustland K. and Hoffmaster A.R. 2009. Detection of pathogenic Leptospira spp. through TaqMan polymerase chain reaction targeting the LipL32 gene. Diagnostic Microbiology and Infectious Disease, 64, 247–255CrossRefPubMedGoogle Scholar
  38. Subharat S., Wilson P.R., Heuer C. and Collins-Emerson J.M. 2011. Evaluation of a SYTO9 real-time polymerase chain reaction assay to detect and identify pathogenic Leptospira species in kidney tissue and urine of New Zealand farmed deer. Journal of Veterinary Diagnostic Investigation, 23, 743–752CrossRefPubMedGoogle Scholar
  39. Talpada M.D., Garvey N., Sprowls R., Eugster A.K. and Vinetz J.M. 2003. Prevalence of leptospiral infection in Texas cattle: implications for transmission to humans. Vector-Borne and Zoonotic Diseases, 3 Suppl 3, 141–147CrossRefPubMedGoogle Scholar
  40. Tourbah A., Fontaine B., Veil N., Mazetti P., Lyon-Caen O. 1992. Lymphocytic meningitis caused by Leptospira panama. La Presse Médicale, 21: 536.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Melissa H. Pinna
    • 1
  • Gabriel Martins
    • 2
  • Ana Paula Loureiro
    • 2
  • Walter Lilenbaum
    • 2
  1. 1.Universidade Federal da BahiaSalvadorBrazil
  2. 2.Laboratório de Bacteriologia VeterináriaUniversidade Federal FluminenseNiteróiBrazil

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