JVAS announces awards - Four best research articles (one each from basic, production, para-clinical and clinical subjects) and one best short communication will be adjudged for awards each year!!!

Journal of Veterinary and Animal Sciences

Volume: 52 Issue: 3

  • Open Access
  • Research Article

Multi-locus sequence typing for species/serovar identification of clinical isolates of Leptospira spp.

D. Divya1**, Siju Joseph1, M. Mini1, R. Sreeja Nair1 and K. Justin Davis2

1. Department of Veterinary Microbiology

2. Department of Veterinary Epidemiology and Preventive Medicine

College of Veterinary and Animal Sciences, Mannuthy, Thrissur- 680 651 Kerala Veterinary and Animal Sciences University, Kerala, India.

**Corresponding author email: [email protected], Ph. 8148676243

Year: 2021, Page: 238-244, Doi: https://doi.org/10.51966/jvas.2021.52.3.238-244

Received: Jan. 18, 2021 Accepted: March 8, 2021 Published: Sept. 30, 2021

Abstract

Leptospirosis is an emerging zoonotic disease endemic in Kerala and close monitoring of emerging serovars is essential to adopt appropriate control strategies. Multi-Locus Sequence Typing (MLST) was reported to be less expensive compared to other cumbersome methods like whole genome sequencing. The present study was conducted to obtain isolates of Leptospira from infected animals and rats and for the identification of serovars using MLST. A total of 205 blood samples (dog, cat, cattle, goat), 43 urine samples (dog, cattle) and post-mortem kidney samples from various animals such as dog (n=12), cattle (n=2) and rat (n=25) were collected and subjected to polymerase chain reaction (PCR) using G1/G2 primers to identify the pathogenic Leptospira. Fifteen samples were found to be positive, these samples when inoculated in the EllinghausenMcCullough-Johnson-Harris (EMJH) semi-solid medium to obtain ten isolates. These ten isolates were further subjected to secY, icdA and GyraseB PCR and sequenced. The obtained sequences were analysed using BLAST and were fed into specified MLST database of Leptospira scheme-3, the allelic profile and sequence type were generated. The MLST results obtained in the study indicated that the isolates S24 and S33 belonged to serovar Canicola, S40 and 47 were Sejroe and S19, S27, S55, S69 and S71 were Bataviae, Autumnalis, Pomona, Icterohaemorraghiae and Australis, respectively. It was concluded that MLST is a convenient method for identifying leptospiral serovars.

Keywords: Leptospirosis, isolation, PCR, MLST.

References

Abhinay, G., Ambily, R. and Joseph, S. 2012. IgM immunoprofile of leptospiral outer membrane proteins in acute canine leptospirosis. Indian Vet. J. 89: 09-10.

Ahmed, N., Devi, S. M., Valverde, M. de los A., Vijayachari, P., Machang’u, R. S., Ellis, W. A. and Hartskeerl, R. A. 2006. Multilocus sequence typing method for identification and genotypic classification of pathogenic Leptospira species. Ann. Clin. Microbiol. Antimicrob. 5: 1-10.

Ahmed, A., Thaipadungpanit, J., Boonsilp, S., Wuthiekanun, V., Nalam, K., Spratt, B. G., Aanensen, D. M., Smythe, L. D., Ahmed, N., Feil, E. J. and Hartskeerl, R. A. 2011. Comparison of two multilocus sequence based genotyping schemes for Leptospira species. PLOS Negl. Trop. Dis. 11: 374-376.

Bourhy, P., Collet, L., Lernout, T., Zinini, F., Hartskeerl, R. A., Van der Linden, H., Thiberge, J. M., Diancourt, L., Brisse, S., Giry, C., Pettinelli, F. and Picardeau, M. 2012. Human leptospira isolates circulating in Mayotte (Indian Ocean) have unique serological and molecular features. J. Clin. Microbiol. 50: 307-311.

Chandran, R. A. 2017. Molecular characterization of leptospiral isolates from domestic animals, rats and human beings. M.V.Sc thesis, Kerala Veterinary and Animal Sciences University, Pookode, 100 p.

Ellis, W. A. and Thiermann, A. B. 1986. Isolation of leptospires from the genital tracts of Iowa cows. Am. J. Vet. Res. 47: 1694- 1696.

Gravekamp, C., van de Kemp, H., Franzen, M., Carrington, D., Schoone, G. J., Van Eys, G. J. J. M., Everard, C. O. R., Hartskeerl, R. A. and Terpstra, W. J. 1993. Detection of seven species of pathogenic leptospires by PCR using two sets of primers. Microbiology. 139: 1691-1700.

Harskeerl, R. A. and Terpstra, W. J. 1996. Leptospirosis in wild animals. Vet. Q. 18: 149-150.

Merien, F., Baranton, G. and Perolat, P. 1995. Comparison of polymerase chain reaction with microagglutination test and culture for diagnosis of leptospirosis. J. Infect. Dis. 172: 281-285.

Natarajaseenivasan, K., Vedhagiri, K., Sivabalan, V., Prabagaran, S. G., Sukumar, S., Artiushin, S. C. and Timoney, J. F. 2011. Seroprevalence of Leptospira borgpetersenii serovar Javanica infection among dairy cattle, rats and humans in the Cauvery river valley of southern India. Southeast Asian J. Trop. Med. Public Hlth. 42: 679-681.

Romero, E. C., Blanco, R. M. and Galloway, R. L. 2011. Analysis of multilocus sequence typing for identification of Leptospira spp. isolates in Brazil. J. Clin. Microbiol. 11: 3940-3942.

Slack, A. T., Symonds, M. L., Dohnt, M. F. and Smythe, L. D. 2006. Identification of pathogenic Leptospira species by conventional or real-time PCR and sequencing of the DNA gyrase subunit B encoding gene. BMC Microbiol. 6: 1471-2180.

Varni, V., Chiani, Y., Nagel, A., Ruybal, P., Vanasco, N. B. and Caimi, K. 2018. Simplified MLST scheme for direct typing of Leptospira human clinical samples. Pathog. Glob. Hlth. 4: 203-209.

Cite this article

Divya, D., Siju Joseph., Mini, M., Sreeja Nair, R. and Justin Davis, K. 2021. Multi-locus sequence typing for species/serovar identification of clinical isolates of Leptospira. J. Vet. Anim. Sci. 52(3): 238-244.

DOI: https://doi.org/10.51966/jvas.2021.52.3.238-244

Views
221
Downloads
71
Citations