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Open Access Research article

Establishment of a leptospirosis model in guinea pigs using an epicutaneous inoculations route

Yan Zhang1, Xiao-Li Lou12, Hong-Liang Yang3, Xiao-Kui Guo1, Xiang-Yan Zhang1, Ping He1* and Xu-Cheng Jiang4*

Author Affiliations

1 Department of Microbiology and Parasitology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

2 Department of Center Laboratory, Songjiang Hospital Affiliated to First People's Hospital, Shanghai Jiao Tong University, Shanghai 201600, China

3 Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523-1682, USA

4 Department of Pathology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

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BMC Infectious Diseases 2012, 12:20  doi:10.1186/1471-2334-12-20

Published: 25 January 2012

Abstract

Background

Leptospires are presumed to enter their host via small abrasions or breaches of the skin. The intraperitoneal route, although commonly used in guinea pig and hamster models of leptospirosis, does not reflect conditions encountered during natural infection. The aim of this study is to develop a novel leptospirosis guinea pig model through epicutaneous route and to elucidate the pathogenesis of leptospirosis in experimental guinea pigs by comparing the data from other studies using different infection routes.

Methods

The guinea pigs were inoculated with 5 × 108 Leptospira interrogans strain Lai onto either shaved-only or abraded skin. The guinea pigs were sacrificed at 2, 8, 24, 48, 72, 96 and 144 h post-infection (p.i.) followed by harvest of the lungs, liver, kidneys, spleen, and the skin around the inoculated sites for further examinations. Hematoxylin and eosin (HE) staining and electron microscopy were used to detect the pathologic changes. Real time PCR and immunohistochemistry staining were performed to detect dynamic distribution of leptospires in blood and tissues, respectively.

Results

In the guinea pigs with abraded skin inoculations, leptospires were detected in blood as early as 2 h post infection (p.i.) and then disseminated to the liver, lungs and kidneys of almost all animals within 96 h p.i.. Leptospires were also detected engulfed in the swelling vascular endothelial cells and were frequently aggregated around the capillaries in the dermis and subcutaneous tissue under the inoculated site. For the guinea pigs with abraded skin inoculations, hemorrhage at the dermis around the inoculated site was found before the appearance of internal organs hemorrhage, severe lesions such as hemorrhages in the lungs, nephritis, jaundice, haematuria were also observed, and two of seven guinea pigs died at 144 h p.i. while no lesions and leptospires were detected in the shaved-only guinea pigs using the same dose of strain Lai.

Conclusion

Intact keratinocyte layer is a very efficient barrier against leptospires, and intact skin can prevent the infiltration of leptosipres to the host. Leptospires can penetrate abraded skin and quickly establish a systemic infection by crossing tissue barriers. We have successfully established a novel leptospirosis guinea pig model through epicutaneous inoculations route, which replicates a natural course of infection and appears to be an alternative way to investigate the pathogenesis of leptospirosis, especially in terms of early stage of host-pathogen interactions. This novel model may also be advantageous for studies of the mechanisms involved in cutaneous barriers and epidermal interactions with this organism.