1887

Abstract

causes cholera outbreaks in endemic regions where the water quality and sanitation facilities remain poor. Apart from biotype and serotype changes, undergoes phase variation, which results in the generation of two morphologically different variants termed smooth and rugose. In this study, 12 rugose (R-VC) and 6 smooth (S-VC) O1 Ogawa isolates were identified in a cholera outbreak that occurred in Hyderabad, India. Antimicrobial susceptibility results showed that all the isolates were resistant to ampicillin, furazolidone and nalidixic acid. In addition, R-VC isolates were resistant to ciprofloxacin (92 %), streptomycin (92 %), erythromycin (83 %), trimethoprim-sulfamethoxazole (75 %) and tetracycline (75 %). Based on the gene analysis, all the isolates were identified as El Tor variant with mutation in two positions of , similar to the classical biotype. The R-VC isolates specifically showed excessive biofilm formation and were comparatively less motile. In addition, the majority of these isolates (~83 %) displayed random mutations in the gene, which encodes haemagglutinin protease regulatory protein. In the PFGE analysis, R-VC and S-VC were placed in distinct clusters but remained clonally related. In the ribotyping analysis, all the R-VC isolates exhibited R-III pattern, which is a prevailing type among the current El Tor isolates. A deletion mutant generated using an S-VC isolate expressed rugose phenotype. To our knowledge, this is the first report on the association of rugose O1 in a large cholera outbreak with extended antimicrobial resistance and random mutations in the haemagglutinin protease regulatory protein encoding gene ().

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000344
2016-10-18
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jmm/65/10/1130.html?itemId=/content/journal/jmm/10.1099/jmm.0.000344&mimeType=html&fmt=ahah

References

  1. Ali A., Mahmud Z. H., Morris J. G., Sozhamannan S., Johnson J. A. 2000; Sequence analysis of TnphoA insertion sites in Vibrio cholerae mutants defective in rugose polysaccharide production. Infect Immun 68:6857–6864 [View Article][PubMed]
    [Google Scholar]
  2. Beyhan S., Bilecen K., Salama S. R., Casper-Lindley C., Yildiz F. H. 2007; Regulation of rugosity and biofilm formation in Vibrio cholerae: comparison of VpsT and VpsR regulons and epistasis analysis of vpsT, vpsR, and hapR . J Bacteriol 189:388–402 [View Article][PubMed]
    [Google Scholar]
  3. CLSI 2010 Performance Standards for Antimicrobial Susceptibility Testing 20th Informational Supplement M100-S20 Wayne, PA: Clinical Laboratory Standards Institute;
    [Google Scholar]
  4. Cooper K. L., Luey C. K., Bird M., Terajima J., Nair G. B., Kam K. M., Arakawa E., Safa A., Cheung D. T. et al. 2006; Development and validation of a PulseNet standardized pulsed-field gel electrophoresis protocol for subtyping of Vibrio cholerae . Foodborne Pathog Dis 3:51–58 [View Article][PubMed]
    [Google Scholar]
  5. Dutta B., Ghosh R., Sharma N. C., Pazhani G. P., Taneja N., Raychowdhuri A., Sarkar B. L., Mondal S. K., Mukhopadhyay A. K. et al. 2006; Spread of cholera with newer clones of Vibrio cholerae O1 El Tor, serotype Inaba, in India. J Clin Microbiol 44:3391–3339 [View Article][PubMed]
    [Google Scholar]
  6. Faruque A. S., Alam K., Malek M. A., Khan M. G., Ahmed S., Saha D., Khan W. A., Nair G. B., Salam M. A. et al. 2007; Emergence of multidrug-resistant strain of Vibrio cholerae O1 in Bangladesh and reversal of their susceptibility to tetracycline after two years. J Health Popul Nutr 25:241–243[PubMed]
    [Google Scholar]
  7. Faruque S. M., Albert M. J., Mekalanos J. J. 1998; Epidemiology, genetics, and ecology of toxigenic Vibrio cholerae . Microbiol Mol Biol Rev 62:1301–1314[PubMed]
    [Google Scholar]
  8. Faruque S. M., Chowdhury N., Kamruzzaman M., Dziejman M., Rahman M. H., Sack D. A., Nair G. B., Mekalanos J. J. 2004; Genetic diversity and virulence potential of environmental Vibrio cholerae population in a cholera-endemic area. Proc Natl Acad Sci U S A 101:2123–2128 [View Article][PubMed]
    [Google Scholar]
  9. Garg P., Sinha S., Chakraborty R., Bhattacharya S. K., Nair G. B., Ramamurthy T., Takeda Y. 2001; Emergence of fluoroquinolone-resistant strains of Vibrio cholerae O1 biotype El Tor among hospitalized patients with cholera in Calcutta, India. Antimicrob Agents Chemother 45:1605–1606 [View Article][PubMed]
    [Google Scholar]
  10. Islam M. S., Jahid M. I., Rahman M. M., Rahman M. Z., Islam M. S., Kabir M. S., Sack D. A., Schoolnik G. K. 2007; Biofilm acts as a microenvironment for plankton-associated Vibrio cholerae in the aquatic environment of Bangladesh. Microbiol Immunol 51:369–379 [View Article][PubMed]
    [Google Scholar]
  11. Lauriano C. M., Ghosh C., Correa N. E., Klose K. E. 2004; The sodium-driven flagellar motor controls exopolysaccharide expression in Vibrio cholerae . J Bacteriol 186:4864–4874 [View Article][PubMed]
    [Google Scholar]
  12. Liang W., Silva A. J., Benitez J. A. 2007; The cyclic AMP receptor protein modulates colonial morphology in Vibrio cholerae . Appl Environ Microbiol 73:7482–7487 [View Article][PubMed]
    [Google Scholar]
  13. Lim B., Beyhan S., Meir J., Yildiz F. H. 2006; Cyclic-diGMP signal transduction systems in Vibrio cholerae: modulation of rugosity and biofilm formation. Mol Microbiol 60:331–348 [View Article][PubMed]
    [Google Scholar]
  14. Lipp E. K., Huq A., Colwell R. R. 2002; Effects of global climate on infectious disease: the cholera model. Clin Microbiol Rev 15:757–770 [View Article][PubMed]
    [Google Scholar]
  15. Mizunoe Y., Wai S. N., Takade A., Yoshida S. I. 1999; Isolation and characterization of rugose form of Vibrio cholerae O139 strain MO10. Infect Immun 67:958–963[PubMed]
    [Google Scholar]
  16. Morita M., Ohnishi M., Arakawa E., Bhuiyan N. A., Nusrin S., Alam M., Siddique A. K., Qadri F., Izumiya H. et al. 2008; Development and validation of a mismatch amplification mutation PCR assay to monitor the dissemination of an emerging variant of Vibrio cholerae O1 biotype El Tor. Microbiol Immunol 52:314–317 [View Article][PubMed]
    [Google Scholar]
  17. Morris J. G., Sztein M. B., Rice E. W., Nataro J. P., Losonsky G. A., Panigrahi P., Tacket C. O., Johnson J. A. 1996; Vibrio cholerae O1 can assume a chlorine-resistant rugose survival form that is virulent for humans. J Infect Dis 174:1364–1368 [View Article][PubMed]
    [Google Scholar]
  18. Nair G. B., Qadri F., Holmgren J., Svennerholm A. M., Safa A., Bhuiyan N. A., Ahmad Q. S., Faruque S. M., Faruque A. S. et al. 2006; Cholera due to altered El Tor strains of Vibrio cholerae O1 in Bangladesh. J Clin Microbiol 44:4211–4213 [View Article][PubMed]
    [Google Scholar]
  19. Pal R. R., Bag S., Dasgupta S., Bhadra R. K. 2012; Functional characterization of the stringent response regulatory gene dksA of Vibrio cholerae and its role in modulation of virulence phenotypes. J Bacteriol 194:5638–5648 [View Article][PubMed]
    [Google Scholar]
  20. Pant A., Anbumani D., Bag S., Mehta O., Kumar P., Saxena S., Nair G. B., Das B. 2015; Effect of LexA on chromosomal integration of CTXϕ in Vibrio cholerae . J Bacteriol 198:268–275 [View Article]
    [Google Scholar]
  21. Popovic T., Bopp C., Olsvik O., Wachsmuth K. 1993; Epidemiologic application of a standardized ribotype scheme for Vibrio cholerae O1. J Clin Microbiol 31:2474–2482[PubMed]
    [Google Scholar]
  22. Rashid M. H., Rajanna C., Ali A., Karaolis D. K. 2003; Identification of genes involved in the switch between the smooth and rugose phenotypes of Vibrio cholerae . FEMS Microbiol Lett 227:113–119 [View Article][PubMed]
    [Google Scholar]
  23. Rashid M. H., Rajanna C., Zhang D., Pasquale V., Magder L. S., Ali A., Dumontet S., Karaolis D. K. 2004; Role of exopolysaccharide, the rugose phenotype and VpsR in the pathogenesis of epidemic Vibrio cholerae . FEMS Microbiol Lett 230:105–113 [View Article][PubMed]
    [Google Scholar]
  24. Ray V. A., Morris A. R., Visick K. L. 2012; A semi-quantitative approach to assess biofilm formation using wrinkled colony development. J Vis Exp 64:e4035 [View Article][PubMed]
    [Google Scholar]
  25. Raychoudhuri A., Mukhopadhyay A. K., Ramamurthy T., Nandy R. K., Takeda Y., Nair G. B. 2008; Biotyping of Vibrio cholerae O1: time to redefine the scheme. Indian J Med Res 128:695–698[PubMed]
    [Google Scholar]
  26. Rice E. W., Johnson C. J., Clark R. M., Fox K. R., Reasoner D. J., Dunnigan M. E., Panigrahi P., Johnson J. A., Morris J. G. 1992; Chlorine and survival of ‘rugose’ Vibrio cholerae . Lancet 340:740 [View Article][PubMed]
    [Google Scholar]
  27. Rice E. W., Johnson C. H., Clark R. M., Fox K. R., Reasoner D. J., Dunnigan M. E., Panigrahi P., Johnson J. A., Morris J. G. Jr 1993; Vibrio cholerae O1 can assume a ‘rugose’ survival form that resists killing by chlorine, yet retains virulence. Int J Environ Health Res 3:89–98 [View Article]
    [Google Scholar]
  28. Sack D. A., Sack R. B., Nair G. B., Siddique A. K. 2004; Cholera. Lancet 363:223–233 [View Article][PubMed]
    [Google Scholar]
  29. Taneja N., Sangar G., Chowdhury G., Ramamurthy T., Mishra A., Singh M., Sharma M. 2012; Molecular epidemiology of Vibrio cholerae causing outbreaks & sporadic cholera in northern India. Indian J Med Res 136:656–663[PubMed]
    [Google Scholar]
  30. Wallace C. K., Anderson P. N., Brown T. C., Khanra S. R., Lewis G. W., Pierce N. F., Sanyal S. N., Segre G. V., Waldman R. H. 1968; Optimal antibiotic therapy in cholera. Bull World Health Organ 39:239–245[PubMed]
    [Google Scholar]
  31. White P. B. 1938; The rugose variant of vibrios. J Pathol Bacteriol 46:1–6 [View Article]
    [Google Scholar]
  32. WHO 1983 Manual for Laboratory Identification of Acute Enteric Infectios Geneva: World Health Organization; WHO/CDD/83.3
    [Google Scholar]
  33. WHO 2015; Cholera annual report 2014. Wkly Epidemiol Rec 90:517–544
    [Google Scholar]
  34. Yildiz F. H., Schoolnik G. K. 1999; Vibrio cholerae O1 El Tor: identification of a gene cluster required for the rugose colony type, exopolysaccharide production, chlorine resistance, and biofilm formation. Proc Natl Acad Sci U S A 96:4028–4033 [View Article][PubMed]
    [Google Scholar]
  35. Yildiz F. H., Liu X. S., Heydorn A., Schoolnik G. K. 2004; Molecular analysis of rugosity in a Vibrio cholerae O1 El Tor phase variant. Mol Microbiol 53:497–515 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.000344
Loading
/content/journal/jmm/10.1099/jmm.0.000344
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error