1887

Abstract

The dynamics of leptospirosis infection have been poorly studied. The purpose of this study was to determine the LD, rate of bacterial dissemination, histopathology and antibody responses against leptospira following inoculation with the highly virulent Fiocruz L1-130 strain in a guinea pig model of leptospirosis. Three routes of infection (intraperitoneal, conjunctival and subcutaneous inoculation) were used to establish disease in guinea pigs. The size and kinetics of leptospiral burdens in the blood and tissues of infected animals were determined over a 1 week course of infection using quantitative real-time PCR (qPCR). Bacteraemia peaked at day 5 post-infection reaching more than 5×10 leptospires ml. The highest spirochaetal load was found in the liver and kidneys, and was associated with alterations in organ tissues and a decline in liver and kidney functions. In contrast, lesions and bacteria were not detected in guinea pigs infected with an avirulent strain derived from a high-passage-number -passaged variant of the Fiocruz L1-130 strain. The use of qPCR supports the findings of earlier studies and provides an easy and reliable method for the quantification of in the tissues of infected animals. qPCR will be used in future studies to evaluate the efficacy of vaccine candidates against leptospirosis and the virulence of selected mutants relative to the parental strain.

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2009-05-01
2019-10-13
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References

  1. Bharti, A. R., Nally, J. E., Ricaldi, J. N., Matthias, M. A., Diaz, M. M., Lovett, M. A., Levett, P. N., Gilman, R. H., Willig, M. R. & other authors ( 2003; ). Leptospirosis: a zoonotic disease of global importance. Lancet Infect Dis 3, 757–771.[CrossRef]
    [Google Scholar]
  2. Bourhy, P., Louvel, H., Saint Girons, I. & Picardeau, M. ( 2005; ). Random insertional mutagenesis of Leptospira interrogans, the agent of leptospirosis, using a mariner transposon. J Bacteriol 187, 3255–3258.[CrossRef]
    [Google Scholar]
  3. Bourne, N., Milligan, G. N., Stanberry, L. R., Stegall, R. & Pyles, R. B. ( 2005; ). Impact of immunization with glycoprotein D2/AS04 on herpes simplex virus type 2 shedding into the genital tract in guinea pigs that become infected. J Infect Dis 192, 2117–2123.[CrossRef]
    [Google Scholar]
  4. Brown, P. D., Carrington, D. G., Gravekamp, C., Van de Kemp, H., Edwards, C. N., Jones, S. R., Prussia, P. R., Garriques, S., Terpstra, W. J. & Levett, P. N. ( 2003; ). Direct detection of leptospiral material in human postmortem samples. Res Microbiol 154, 581–586.[CrossRef]
    [Google Scholar]
  5. Cox, J. S., Chen, B., McNeil, M. & Jacobs, W. R. J. ( 1999; ). Complex lipid determines tissue-specific replication of Mycobacterium tuberculosis in mice. Nature 402, 79–83.[CrossRef]
    [Google Scholar]
  6. Ellinghausen, H. C. & McCullough, W. G. ( 1965; ). Nutrition of Leptospira pomona and growth of 13 other serotypes: fractionation of oleic albumin complex and a medium of bovine albumin and polysorbate 80. Am J Vet Res 26, 45–51.
    [Google Scholar]
  7. Faine, S. ( 1957a; ). Virulence in Leptospira. I. Reactions of guinea-pigs to experimental infections with Leptospira icterohaemorrhagiae. Br J Exp Pathol 38, 1–7.
    [Google Scholar]
  8. Faine, S. ( 1957b; ). Virulence in Leptospira. II. The growth in vivo of virulent Leptospira icterohaemorrhagiae. Br J Exp Pathol 38, 8–14.
    [Google Scholar]
  9. Johnson, R. C. & Harris, V. G. ( 1967; ). Differentiation of pathogenic and saprophytic leptospires. J Bacteriol 94, 27–31.
    [Google Scholar]
  10. Ko, A. I., Galvao Reis, M., Ribeiro Dourado, C. M., Johnson, W. D. J. & Riley, L. W. ( 1999; ). Urban epidemic of severe leptospirosis in Brazil. Salvador Leptospirosis Study Group. Lancet 354, 820–825.[CrossRef]
    [Google Scholar]
  11. Levett, P. N. ( 2001; ). Leptospirosis. Clin Microbiol Rev 14, 296–326.[CrossRef]
    [Google Scholar]
  12. Masuzawa, T., Hashiguchi, Y., Nakamura, R., Suzuki, R., Shimizu, T., Iwamoto, Y., Morita, T. & Yanagihara, Y. ( 1991; ). Experimental lethal infection of Leptospira interrogans in mice treated with cyclophosphamide. Can J Microbiol 37, 312–315.[CrossRef]
    [Google Scholar]
  13. McBride, A. J., Athanazio, D. A., Reis, M. G. & Ko, A. I. ( 2005; ). Leptospirosis. Curr Opin Infect Dis 18, 376–386.[CrossRef]
    [Google Scholar]
  14. Mérien, F., Amouriaux, P., Perolat, P., Baranton, G. & Saint Girons, I. ( 1992; ). Polymerase chain reaction for detection of Leptospira spp. in clinical samples. J Clin Microbiol 30, 2219–2224.
    [Google Scholar]
  15. Murray, G. L., Morel, V., Cerqueira, G. M., Croda, J., Srikram, A., Henry, R., Ko, A. I., Dellagostin, O. A., Bulach, D. M. & other authors ( 2008; ). Genome-wide transposon mutagenesis in pathogenic Leptospira spp. Infect Immun 77, 810–816.
    [Google Scholar]
  16. Nally, J. E., Fishbein, M. C., Blanco, D. R. & Lovett, M. A. ( 2005; ). Lethal infection of C3H/HeJ and C3H/SCID mice with an isolate of Leptospira interrogans serovar Copenhageni. Infect Immun 73, 7014–7017.[CrossRef]
    [Google Scholar]
  17. Nascimento, A. L., Ko, A. I., Martins, E. A., Monteiro-Vitorello, C. B., Ho, P. L., Haake, D. A., Verjovski-Almeida, S., Hartskeerl, R. A., Marques, M. V. & other authors ( 2004; ). Comparative genomics of two Leptospira interrogans serovars reveals novel insights into physiology and pathogenesis. J Bacteriol 186, 2164–2172.[CrossRef]
    [Google Scholar]
  18. Pereira, M. M., Andrade, J., Marchevsky, R. S. & Ribeiro dos Santos, R. ( 1998; ). Morphological characterization of lung and kidney lesions in C3H/HeJ mice infected with Leptospira interrogans serovar icterohaemorrhagiae: defect of CD4+ and CD8+ T-cells are prognosticators of the disease progression. Exp Toxicol Pathol 50, 191–198.[CrossRef]
    [Google Scholar]
  19. Pereira, M. M., Da Silva, J. J., Pinto, M. A., Da Silva, M. F., Machado, M. P., Lenzi, H. L. & Marchevsky, R. S. ( 2005; ). Experimental leptospirosis in marmoset monkeys (Callithrix jacchus): a new model for studies of severe pulmonary leptospirosis. Am J Trop Med Hyg 72, 13–20.
    [Google Scholar]
  20. Reed, L. J. & Muench, H. A. ( 1938; ). Simple method of determining fifty percent endpoints. Am J Hyg 27, 494–497.
    [Google Scholar]
  21. Ristow, P., Bourhy, P., Da Cruz McBride, F. W., Figueira, C. P., Huerre, M., Ave, P., Girons, I. S., Ko, A. I. & Picardeau, M. ( 2007; ). The OmpA-like protein Loa22 is essential for leptospiral virulence. PLoS Pathog 3, e97 [CrossRef]
    [Google Scholar]
  22. Roczek, A., Forster, C., Raschel, H., Hörmansdorfer, S., Bogner, K. H., Hafner-Marx, A., Lepper, H., Dobler, G., Büttner, M. & Sing, A. ( 2008; ). Severe course of rat bite-associated Weil's disease in a patient diagnosed with a new Leptospira-specific real-time quantitative LUX-PCR. J Med Microbiol 57, 658–663.[CrossRef]
    [Google Scholar]
  23. Romero, E. C., Billerbeck, A. E., Lando, V. S., Camargo, E. D., Souza, C. C. & Yasuda, P. H. ( 1998; ). Detection of Leptospira DNA in patients with aseptic meningitis by PCR. J Clin Microbiol 36, 1453–1455.
    [Google Scholar]
  24. Segura, E. R., Ganoza, C. A., Campos, K., Ricaldi, J. N., Torres, S., Silva, H., Céspedes, M. J., Matthias, M. A, Swancutt, M. A. & other authors ( 2005; ). Clinical spectrum of pulmonary involvement in leptospirosis in a region of endemicity, with quantification of leptospiral burden. Clin Infect Dis 40, 343–351.[CrossRef]
    [Google Scholar]
  25. Silva, E. F., Santos, C. S., Athanazio, D. A., Seyffert, N., Seixas, F. K., Cerqueira, G. M, Fagundes, M. Q., Brod, C. S., Reis, M. G. & other authors ( 2008; ). Characterization of virulence of Leptospira isolates in a hamster model. Vaccine 26, 3892–3896.[CrossRef]
    [Google Scholar]
  26. Smythe, L. D., Smith, I. L., Smith, G. A., Dohnt, M. F., Symonds, M. L., Barnett, L. J. & McKay, D. B. ( 2002; ). A quantitative PCR (TaqMan) assay for pathogenic Leptospira spp. BMC Infect Dis 2, 13 [CrossRef]
    [Google Scholar]
  27. Sonrier, C., Branger, C., Michel, V., Ruvoën-Clouet, N., Ganière, J. P. & André-Fontaine, G. ( 2000; ). Evidence of cross-protection within Leptospira interrogans in an experimental model. Vaccine 19, 86–94.[CrossRef]
    [Google Scholar]
  28. Truccolo, J., Serais, O., Merien, F. & Perolat, P. ( 2001; ). Following the course of human leptospirosis: evidence of a critical threshold for the vital prognosis using a quantitative PCR assay. FEMS Microbiol Lett 204, 317–321.[CrossRef]
    [Google Scholar]
  29. Truccolo, J., Charavay, F., Merien, F. & Perolat, P. ( 2002; ). Quantitative PCR assay to evaluate ampicillin, ofloxacin, and doxycycline for treatment of experimental leptospirosis. Antimicrob Agents Chemother 46, 848–853.[CrossRef]
    [Google Scholar]
  30. Van den Ingh, T. S. & Hartman, E. G. ( 1986; ). Pathology of acute Leptospira interrogans serotype Icterohaemorrhagiae infection in the Syrian hamster. Vet Microbiol 12, 367–376.[CrossRef]
    [Google Scholar]
  31. Viriyakosol, S., Matthias, M. A., Swancutt, M. A., Kirkland, T. N. & Vinetz, J. M. ( 2006; ). Toll-like receptor 4 protects against lethal Leptospira interrogans serovar Icterohaemorrhagiae infection and contributes to in vivo control of leptospiral burden. Infect Immun 74, 887–895.[CrossRef]
    [Google Scholar]
  32. WHO ( 1999; ). Leptospirosis worldwide, 1999. Wkly Epidemiol Rec 74, 237–242.
    [Google Scholar]
  33. Yang, H. L., Jiang, X. C., Zhang, X. Y., Li, W. J., Hu, B. Y., Zhao, G. P. & Guo, X. K. ( 2006; ). Thrombocytopenia in the experimental leptospirosis of guinea pig is not related to disseminated intravascular coagulation. BMC Infect Dis 6, 19 [CrossRef]
    [Google Scholar]
  34. Yukawa, M., Mochizuki, K. & Imamura, S. ( 1990; ). Susceptibility of Mongolian gerbils (Meriones unguiculatus) to leptospires and the protective effect of vaccination. Vet Microbiol 24, 63–71.[CrossRef]
    [Google Scholar]
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