Isolation and Molecular Characterization of Leptospira interrogans and Leptospira borgpetersenii Isolates from the Urban Rat Populations of Kuala Lumpur, Malaysia

Douadi Benacer Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia; Laboratory of Biomedical Science and Molecular Microbiology, Institute of Graduate Studies, University of Malaya, Kuala Lumpur, Malaysia; Bacteriology Unit, Institute for Medical Research, Ministry of Health, Kuala Lumpur, Malaysia; Bacterial Zoonoses Special Pathogens Branch, Centers for Disease Control and Prevention, Atlanta, Georgia

Search for other papers by Douadi Benacer in
Current site
Google Scholar
PubMed
Close
,
Siti Nursheena Mohd Zain Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia; Laboratory of Biomedical Science and Molecular Microbiology, Institute of Graduate Studies, University of Malaya, Kuala Lumpur, Malaysia; Bacteriology Unit, Institute for Medical Research, Ministry of Health, Kuala Lumpur, Malaysia; Bacterial Zoonoses Special Pathogens Branch, Centers for Disease Control and Prevention, Atlanta, Georgia

Search for other papers by Siti Nursheena Mohd Zain in
Current site
Google Scholar
PubMed
Close
,
Fairuz Amran Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia; Laboratory of Biomedical Science and Molecular Microbiology, Institute of Graduate Studies, University of Malaya, Kuala Lumpur, Malaysia; Bacteriology Unit, Institute for Medical Research, Ministry of Health, Kuala Lumpur, Malaysia; Bacterial Zoonoses Special Pathogens Branch, Centers for Disease Control and Prevention, Atlanta, Georgia

Search for other papers by Fairuz Amran in
Current site
Google Scholar
PubMed
Close
,
Renee L. Galloway Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia; Laboratory of Biomedical Science and Molecular Microbiology, Institute of Graduate Studies, University of Malaya, Kuala Lumpur, Malaysia; Bacteriology Unit, Institute for Medical Research, Ministry of Health, Kuala Lumpur, Malaysia; Bacterial Zoonoses Special Pathogens Branch, Centers for Disease Control and Prevention, Atlanta, Georgia

Search for other papers by Renee L. Galloway in
Current site
Google Scholar
PubMed
Close
, and
Kwai Lin Thong Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia; Laboratory of Biomedical Science and Molecular Microbiology, Institute of Graduate Studies, University of Malaya, Kuala Lumpur, Malaysia; Bacteriology Unit, Institute for Medical Research, Ministry of Health, Kuala Lumpur, Malaysia; Bacterial Zoonoses Special Pathogens Branch, Centers for Disease Control and Prevention, Atlanta, Georgia

Search for other papers by Kwai Lin Thong in
Current site
Google Scholar
PubMed
Close
Restricted access

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.

Author Notes

* Address correspondence to Kwai Lin Thong, Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur 50603, Malaysia. E-mail: thongkl@um.edu.my

Financial support: This work was supported by University of Malaya Research Grant RG053/11BIO.

Authors' addresses: Douadi Benacer and Kwai Lin Thong, Institute of Biological Sciences, Faculty of Science and Laboratory of Biomedical Science and Molecular Microbiology, Institute of Graduate Studies, University of Malaya, Kuala Lumpur, Malaysia, E-mails: benacer_dvt@hotmail.com and thongkl@um.edu.my. Siti Nursheena Mohd Zain, Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia, E-mail: nsheena@um.edu.my. Fairuz Amran, Bacteriology Unit, Institute for Medical Research, Ministry of Health, Kuala Lumpur, Malaysia, E-mail: fairuz@imr.gov.my. Renee L. Galloway, Bacterial Zoonoses Special Pathogens Branch, Centers for Disease Control and Prevention, Atlanta, GA, E-mail: zul0@cdc.gov.

  • 1.

    Palaniappan RU, Ramanujam S, Chang YF, 2007. Leptospirosis: pathogenesis, immunity, and diagnosis. Curr Opin Infect Dis 20: 284292.

  • 2.

    Hartskeerl RA, Collares-Pereira M, Ellis WA, 2011. Emergence, control and re-emerging leptospirosis: dynamics of infection in the changing world. Clin Microbiol Infect 17: 494501.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 3.

    Socolovschi C, Angelakis E, Renvoisé A, Fournier PE, Marie JL, Davoust B, Stein A, Raoult D, 2011. Strikes, flooding, rats, and leptospirosis in Marseille, France. Int J Infect Dis 15: 710715.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 4.

    Valverde ML, Ramirez JM, Montes de Oca LG, Goris MG, Ahmed N, Hartskeerl RA, 2008. Arenal, a new Leptospira serovar of serogroup Javanica, isolated from a patient in Costa Rica. Infect Genet Evol 8: 529533.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 5.

    Green-Mckenzie J, Shoff WH, 2012. Leptospirosis in Emergency Medicine. E Medicine (WebMD). Available at: http://emedicine.medscape.com/article/788751-overview. Accessed August 15, 2012.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 6.

    Turk N, Milas Z, Margaletic J, Staresina V, Slavica A, Riquelme-Sertour N, Bellenger E, Baranton G, Postic D, 2003. Molecular characterisation of Leptospira spp. strains isolated from small rodents in Croatia. Epidemiol Infect 130: 159166.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 7.

    Bharti AR, Nally JE, Ricaldi JN, Matthias MA, Diaz MM, Lovett MA, Levett PN, Gilman RH, Willig MR, Gotuzzo E, Vinetz JM; Peru-United States Leptospirosis Consortium, 2003. Leptospirosis: a zoonotic disease of global importance. Lancet Infect Dis 3: 757771.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 8.

    Victoriano AF, Smythe LD, Gloriani-Barzaga N, Cavinta LL, Kasai T, Limpakarnjanarat K, Ong BL, Gongal G, Hall J, Coulombe CA, Yanagihara Y, Yoshida S, Adler B, 2009. Leptospirosis in the Asia Pacific region. BMC Infect Dis 9: 147.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 9.

    Dikken H, Kmety E, 1978. Serological typing methods of leptospires. Bergan T, Norris JR, eds. Methods in Microbiology. London, UK: Academic Press, 259307.

  • 10.

    Levett PN, 2001. Leptospirosis. Clin Microbiol Rev 14: 296326.

  • 11.

    Bal AE, Gravekamp C, Hartskeerl RA, De Meza-Brewster J, Korver H, Terpstra WJ, 1994. Detection of leptospires in urine by PCR for early diagnosis of leptospirosis. J Clin Microbiol 32: 18941898.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 12.

    Fonseca C de A, Teixeira MM, Romero EC, Tengan FM, Silva MV, Shikanai-Yasuda MA, 2006. Leptospira DNA detection for the diagnosis of human leptospirosis. J Infect 52: 1522.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 13.

    Bomfim MR, Barbosa-Stancioli EF, Koury MC, 2008. Detection of pathogenic leptospires in urine from naturally infected cattle by nested PCR. Vet J 178: 251256.

  • 14.

    Vital-Brazil JM, Balassiano IT, Oliveira FS, Costa AD, Hillen L, Pereira MM, 2010. Multiplex PCR-based detection of Leptospira in environmental water samples obtained from a slum settlement. Mem Inst Oswaldo Cruz 105: 353355.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 15.

    Galloway RL, Levett PN, 2008. Evaluation of a modified pulsed-field gel electrophoresis approach for the identification of Leptospira serovars. Am J Trop Med Hyg 78: 628632.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 16.

    Galloway RL, Levett PN, 2010. Application and validation of PFGE for serovar identification of Leptospira clinical isolates. PLoS Negl Trop Dis 4: e824.

  • 17.

    Levett PN, 2007. Sequence-based typing of Leptospira: epidemiology in the genomic era. PLoS Negl Trop Dis 1: e120.

  • 18.

    Hartskeerl RA, 2006. Leptospirosis: current status and future trends. Indian J Med Microbiol 24: 309.

  • 19.

    Lokman HS, 2011. Guidelines for the Diagnosis, Management, Prevention and Control of Leptospirosis in Malaysia, 1st Ed. Disease Control Division, Department of Public Health, Ministry of Public Health, Malaysia.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 20.

    Bahaman AR, Ibrahim AL, 1988. A review of leptospirosis in Malaysia. Vet Res Commun 12: 179189.

  • 21.

    Koay TK, Nirmal S, Noitie L, Tan E, 2004. An epidemiological investigation of an outbreak of leptospirosis associated with swimming, Beaufort, Sabah. Med J Malaysia 59: 455459.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 22.

    Medway L, 1983. The Wild Mammals of Malaya (Peninsula Malaysia) and Singapore, 2nd Ed. Oxford, UK: Oxford University Press, 131.

  • 23.

    Merien 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: 22192224.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 24.

    Gravekamp C, Van de Kemp H, Franzen M, Carrington D, Schoone GJ, Van Eys GJ, Everard CO, Hartskeerl RA, Terpstra WJ, 1993. Detection of seven species of pathogenic leptospires by PCR using two sets of primers. J Gen Microbiol 139: 16911700.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 25.

    World Health Organization, 2003. Human Leptospirosis: Guidance for Diagnosis, Surveillance and Control. Geneva: World Health Organization and International Leptospirosis Society.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 26.

    Mohd Zain SN, Behnke JM, Lewis JW, 2012. Helminth communities from two urban rat populations in Kuala Lumpur, Malaysia. Parasit Vectors 5: 47.

  • 27.

    Corbet GB, Harris S, 1991. The Handbook of British Mammals. Oxford Blackwell, UK, 588.

  • 28.

    Gordon-Smith CE, Turner LH, Harrison JL, Broom JC, 1961. Animal leptospirosis in Malaya: 1. methods, zoogeographical background, and broad analysis of results. Bull World Health Organ 24: 521.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 29.

    Mohamed-Hassan SN, Bahaman AR, Mutalib AR, Khairani-Bejo S, 2010. Serological prevalence of leptospiral infection in wild rats at the National Service Training Centres in Kelantan and Terenggaanu. Trop Biomed 27: 3032.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 30.

    Mohamed-Hassan SN, Bahaman AR, Mutalib AR, Khairani-Bejo S, 2012. Prevalence of pathogenic leptospires in rats from selected locations in peninsular Malaysia. Res J Anim Sci 6: 1225.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 31.

    Ridzlan FR, Bahaman AR, Khairani-Bejo S, Mutalib AR, 2010. Detection of pathogenic Leptospira from selected environment in Kelantan and Terengganu, Malaysia. Trop Biomed 27: 632638.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 32.

    Faria MT, Calderwood MS, Athanazio DA, McBride AJA, 2008. Carriage of Leptospira interrogans among domestic rats from an urban setting highly endemic for leptospirosis in Brazil. Acta Trop 108: 15.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 33.

    Khairani-Bejo S, Oii SS, Bahaman AR, 2004. Rats: leptospirosis reservoir in Serdang Selangor residential area. J Anim Vet Adv 3: 6669.

  • 34.

    Villanueva SY, Ezoe H, Baterna RA, Yanagihara Y, Muto M, Koizumi N, Fukui T, Okamoto Y, Masuzawa T, Cavinta LL, Gloriani NG, Yoshida S, 2010. Serologic and molecular studies of Leptospira and leptospirosis among rats in the Philippines. Am J Trop Med Hyg 82: 889898.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 35.

    Krøjgaard LH, Villumsen S, Markussen MDK, Jensen JS, Leirs H, Heiberg AC, 2009. High prevalence of Leptospira spp. in sewer rats (Rattus norvegicus). Epidemiol Infect 137: 15861592.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 36.

    Perez J, Brescia F, Becam J, Mauron C, Goarant C, 2011. Rodent abundance dynamics and leptospirosis carriage in an area of hyper-endemicity in New Caledonia. PLoS Negl Trop Dis 5: e1361.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 37.

    Carter ME, Cordes DO, 1980. Leptospirosis and other infections of Rattus rattus and Rattus norvegicus. N Z Vet J 28: 4550.

  • 38.

    Easterbrook JD, Kaplan JB, Vanasco NB, Reeves WK, Purcell RH, Kosoy MY, Glass GE, Watson J, Klein SL, 2007. A survey of zoonotic pathogens carried by Norway rats in Baltimore, Maryland, USA. Epidemiol Infect 135: 11921199.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 39.

    Agudelo-Florez P, Londono AF, Quiroz VH, Angel JC, Moreno N, Loaiza ET, Munoz LF, Rodas JD, 2009. Prevalence of Leptospira spp. in urban rodents from a groceries trade center of Medellin, Colombia. Am J Trop Med Hyg 81: 906910.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 40.

    Wagenaar J, Zuerner RL, Alt D, Bolin CA, 2000. Comparison of polymerase chain reaction assays with bacteriologic culture, immunofluorescence, and nucleic acid hybridization for detection of Leptospira borgpetersenii serovar hardjo in urine of cattle. Am J Vet Res 61: 316320.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 41.

    Fornazari F, Costa da Silva R, Richini-Pereira VB, Beserra HEO, Luvizotto MCR, Langoni H, 2012. Comparison of conventional PCR, quantitative PCR, bacteriological culture and the Warthin Starry technique to detect Leptospira spp. in kidney and liver samples from naturally infected sheep from Brazil. J Microbiol Methods 90: 321326.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 42.

    Shafei MN, Sulong MR, Yaacob NA, Hassan H, Wan Mohd Zahiruddin WM, Daud A, Ismail Z, Abdullah MR, 2012. Seroprevalence of leptospirosis among town service workers on northeastern State of Malaysia. International Journal of Collaborative Research on Internal Medicine & Public Health (IJCRIMPH) 4: 395403.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 43.

    Vedhagiri K, Natarajaseenivasan K, Prabhakaran SG, Selvin J, Narayanan R, Shouche YS, Vijayachari P, Ratnam S, 2010. Characterization of Leptospira borgpetersenii isolates from field rats (Rattus norvegicus) by 16S rRNA and lipL32 gene sequencing. Braz J Microbiol 41: 150157.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 44.

    Romero EC, Blanco RM, Galloway RL, 2009. Application of pulsed-field gel electrophoresis for the discrimination of leptospiral isolates in Brazil. Lett Appl Microbiol 48: 623627.

    • PubMed
    • Search Google Scholar
    • Export Citation
Past two years Past Year Past 30 Days
Abstract Views 374 374 69
Full Text Views 704 27 1
PDF Downloads 338 21 1
 
Membership Banner
 
 
 
Affiliate Membership Banner
 
 
Research for Health Information Banner
 
 
CLOCKSS
 
 
 
Society Publishers Coalition Banner
Save