Title:	Real-Time PCR Reveals Rapid Dissemination of Leptospira interrogans after Intraperitoneal and Conjunctival Inoculation of Hamsters
Authors:	Wunder Júnior, Elsio Augusto Figueira, Claudio Pereira Santos, Gisele R Lourdault, Kristel Matthias, Michael A Vinetz, Joseph M Ramos, Eduardo Antônio Gonçalves Haake, David A Picardeau, Mathieu Reis, Mitermayer Galvão dos Ko, Albert Icksang
Affilliation:	Yale School of Public Health. Epidemiology of Microbial Disease Division. New Haven, Connecticut, USA / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil Institut Pasteur. Unité de Biologie des Spirochètes. Paris, France / University of California. David Geffen School of Medicine. Department of Medicine. Los Angeles, Los Angeles, California, USA University of California San Diego School of Medicine. Division of Infectious Diseases. Department of Medicine. La Jolla, California, USA University of California San Diego School of Medicine. Division of Infectious Diseases. Department of Medicine. La Jolla, California, USA Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil University of California. David Geffen School of Medicine. Department of Medicine. Los Angeles, Los Angeles, California, USA Institut Pasteur. Unité de Biologie des Spirochètes. Paris, France Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil Yale School of Public Health. Epidemiology of Microbial Disease Division. New Haven, Connecticut, USA / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
Abstract:	The pathogen Leptospira interrogans is a highly motile spirochete that causes acute and fulminant infections in humans and other accidental hosts. Hematogenous dissemination is important for infection by the pathogen but remains poorly understood because few animal model studies have used sensitive tools to quantify the bacteria. We evaluated the kinetics of leptospiral infection in Golden Syrian hamsters by a sensitive quantitative real-time PCR (TaqMan) with lipl32 as the target gene. The dissemination and bacterial burden were measured after intraperitoneal infection with a high dose (10(8)) or low dose (2.5 × 10(2)) of leptospires. We also examined the conjunctival challenge route to mimic the natural history of infection. Quantification of leptospires in perfused animals revealed that pathogens were detected in all organs of intraperitoneally infected hamsters, including the eye and brain, within 1 h after inoculation of 10(8) virulent L. interrogans bacteria. Peaks of 10(5) to 10(8) leptospires per gram or per milliliter were achieved in blood and all tissues between day 4 and day 8 after intraperitoneal inoculation of high- and low-dose challenges, respectively, coinciding with macroscopic and histological changes. The conjunctival route resulted in a delay in the time to peak organ burden in comparison to intraperitoneal infection, indicating that although infection could be established, penetration efficiency was low across this epithelial barrier. Surprisingly, infection with a large inoculum of high-passage-number attenuated L. interrogans strains resulted in dissemination to all organs in the first 4 days postinfection, albeit with a lower burden, followed by clearance from the blood and organs 7 days postinfection and survival of all animals. These results demonstrate that leptospiral dissemination and tissue invasion occur. In contrast, development of a critical level of tissue burden and pathology are dependent on the virulence of the infecting strain.
Keywords:	Leptospira interrogans Leptospirosis Infection Hamsters Animals
keywords:	Leptospira interrogans Leptospirose Infecção Hamsters Animais
Issue Date:	2016
Publisher:	American Society for Microbiology
Citation:	WUNDER JÚNIOR, E. A. et al. Real-Time PCR Reveals Rapid Dissemination of Leptospira interrogans after Intraperitoneal and Conjunctival Inoculation of Hamsters. Infection and Immunity, v. 84, n. 7, p. 2105-2115, 2016.
DOI:	10.1128/IAI.00094-16
ISSN:	0019-9567
Copyright:	open access
Appears in Collections:	BA - IGM - Artigos de Periódicos

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