Profiling of Leptospira interrogans, L-santarosai, L-meyeri and L-borgpetersenii by SE-AFLP, PFGE and susceptibility testing-a continuous attempt at species and serovar differentiation

Author(s)

Moreno, Luisa Z. ^[1] ; Miraglia, Fabiana ^{[2, 1]} ; Lilenbaum, Walter ^[2] ; Neto, Jose S. F. ^[1] ; Freitas, Julio C. ^[3] ; Morais, Zenaide M. ^[1] ; Hartskeerl, Rudy A. ^{[4, 5]} ; da Costa, Barbara L. P. ^[1] ; Vasconcellos, Silvio A. ^[1] ; Moreno, Andrea M. ^[1]

Affiliation

^[1] Univ Sao Paulo, Sch Vet Med & Anim Sci, Lab Bacterial Zoonosis, Lab Mol Epidemiol & Antimicrobial Resistance, BR-05508270 Sao Paulo, SP - Brazil

^[2] Univ Fed Fluminense, Dept Microbiol & Parasitol, Vet Bacteriol Lab, Hernani Mello 101, BR-24210130 Niteroi, RJ - Brazil

^[3] Univ Estadual Londrina, Lab Anim Leptospirosis, Rod Celso Garcia Cid, BR-86057970 Londrina, PR - Brazil

^[4] WHO FAO OIE, NL-391105 Amsterdam - Netherlands

^[5] Royal Trop Inst, KIT Biomed Res, Natl Leptospirosis Reference Ctr, NL-391105 Amsterdam - Netherlands

• Document type: Journal article
• Source: EMERGING MICROBES & INFECTIONS; v. 5, MAR 9 2016.
• Web of Science Citations: 5

Abstract

Leptospirosis is a widespread systemic zoonosis, considered as reemerging in certain developing countries. Although the cross agglutinin absorption test is still considered the standard method for Leptospira identification, it presents several disadvantages. The aim of this study was to characterize Leptospira spp. isolated from various hosts by genotyping and broth microdilution susceptibility testing in an attempt to differentiate Leptospira species, serogroups and serovars. Forty-seven isolates were studied. They were previously serotyped, and species confirmation was performed by 16S rRNA sequencing. Single-enzyme amplified fragment length polymorphism (SE-AFLP) and pulsed-field gel electrophoresis (PFGE) analysis enabled the distinction of L. interrogans from L. santarosai, L. meyeri and L. borgpetersenii in two main clusters. Among L. interrogans, it was possible to differentiate into two new clusters the serogroup Icterohaemorrhagiae from the serogroups Canicola and Pomona. L. santarosai isolates presented higher genetic variation than the other species in both techniques. Interestingly, the minimum inhibitory concentration (MIC) cluster analysis also provided Leptospira serogroup differentiation. Further studies are necessary regarding serovar Bananal isolates, as they presented the highest MIC values for most of the antimicrobials tested. All studied techniques successfully distinguished Leptospira species and serogroups. Despite being library-dependent methods, these approaches are less labor intensive and more economically viable, particularly SE-AFLP, and can be implemented in most reference laboratories worldwide to enable faster Leptospira typing. (AU)

• FAPESP's process: 11/18290-0 - Characterization of Leptospira strains isolated in Brazil through multiloccus sequence typing (MLST)
• Grantee: Andrea Micke Moreno
• Support Opportunities: Regular Research Grants
• FAPESP's process: 13/17136-2 - Characterization of Leptospira strains isolated in Brazil by pulsed field gel electrophoresis (PFGE) typing and comparative genomic analysis.
• Grantee: Luisa Zanolli Moreno
• Support Opportunities: Scholarships in Brazil - Doctorate