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. 2013 Apr;88(4):704-9.
doi: 10.4269/ajtmh.12-0662. Epub 2013 Jan 28.

Isolation and molecular characterization of Leptospira interrogans and Leptospira borgpetersenii isolates from the urban rat populations of Kuala Lumpur, Malaysia

Douadi Benacer  1 , Siti Nursheena Mohd Zain, Fairuz Amran, Renee L Galloway, Kwai Lin Thong
Affiliations

Isolation and molecular characterization of Leptospira interrogans and Leptospira borgpetersenii isolates from the urban rat populations of Kuala Lumpur, Malaysia

Douadi Benacer et al. Am J Trop Med Hyg. 2013 Apr.

Abstract

Rats are considered the principal maintenance hosts of Leptospira. The objectives of this study were isolation and identification of Leptospira serovars circulating among urban rat populations in Kuala Lumpur. Three hundred urban rats (73% Rattus rattus and 27% R. norvegicus) from three different sites were trapped. Twenty cultures were positive for Leptospira using dark-field microscopy. R. rattus was the dominant carrier (70%). Polymerase chain reaction (PCR) confirmed that all isolates were pathogenic Leptospira species. Two Leptospira serogroups, Javanica and Bataviae, were identified using microscopic agglutination test (MAT). Pulsed-field gel electrophoresis (PFGE) identified two serovars in the urban rat populations: L. borgpetersenii serovar Javanica (85%) and L. interrogans serovar Bataviae (15%). We conclude that these two serovars are the major serovars circulating among the urban rat populations in Kuala Lumpur. Despite the low infection rate reported, the high pathogenicity of these serovars raises concern of public health risks caused by rodent transmission of leptospirosis.

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Figures

Figure 1.
Figure 1.
Representative gel of PCR for detection of Leptospira genus using LA/LB primers. Lanes 1–8: DNA marker (100 bp); positive control; R2 (urine sample); R18 (urine sample); negative control; R71 (kidney sample); R77 (urine sample); R108 (kidney sample).
Figure 2.
Figure 2.
Representative gel of PCR for confirmation of pathogenic Leptospira using G1/G2 primers. Lanes 1–8: DNA marker (100 bp); positive control; R77 (urine sample); R78 (urine sample); R84 (urine sample); negative control; R102 (urine sample); R108 (kidney sample).
Figure 3.
Figure 3.
PFGE patterns of NotI digested chromosomal DNA of Leptospira isolates from urban rats of Kuala Lumpur. Lanes 1, 6, 11: XbaI digested chromosomal DNA of Salmonella H9812 marker strain; lanes: 2–5, 7–10: L. interrogans serovar Javanica R2; L. interrogans serovar Javanica R18; L. interrogans serovar Javanica R7; L. interrogans serovar Javanica R78; L. interrogans serovar Javanica R84; L. interrogans serovar Bataviae R108; L. interrogans serovar Bataviae R149; L. interrogans serovar Bataviae R183.
Figure 4.
Figure 4.
Dendrogram based on cluster analysis of the PFGE profiles of NotI digested chromosomal Leptospira isolated from urban rats generated using BioNumerics Version 6.0 (Applied Maths, Belgium) software and unweighted pair group arithmetic means methods (UPGMA). Cluster A represents L. interrogans serovar Bataviae, cluster B represents L. borgpetersenii serovar Javanica.
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