|Year : 2014 | Volume
| Issue : 4 | Page : 254-257
Vibrio isolates from cases of acute diarrhea and their antibiotic susceptibility pattern in a tertiary care hospital of Punjab
Atul Kumar, Aroma Oberoi
Department of Microbiology, Christian Medical College, Ludhiana, Punjab, India
|Date of Web Publication||16-Oct-2014|
Dr. Aroma Oberoi
Department of Clinical Microbiology, Christian Medical College, Ludhiana - 141 008, Punjab
Source of Support: None, Conflict of Interest: None
Introduction: Vibrio cholerae, the cause of cholera, is one of the leading causes of morbidity and mortality in many developing countries. Especially during monsoon season, cholera outbreaks have been frequently reported from one or the other part of this country. However, occasional sporadic cases have also been reported, majorly due to lack of proper sanitation and impure water supply. Aim: To determine the prevalence of serotypes of Vibrio isolates from cases of acute diarrhea and their antibiotic susceptibility pattern in a tertiary care hospital, North India. Materials and Methods: A retrospective study was conducted over a period of 1 year from January 2013 to December 2013 in the department of microbiology, in a tertiary care hospital in Punjab. The diagnosis was suggested by the clinical picture and confirmed by stool culture. The stool samples received in the laboratory were processed by standard microbiological techniques for identification of V. cholerae. The suspected colonies of Vibrio were identified by standard biochemical tests and serotyping was done by group specific antisera. The susceptibility of all the isolated Vibrio species to different antibiotics were done by Kirby-Bauer's disk diffusion technique as per the Clinical and Laboratory Standards Institute (CLSI) guidelines. Results: V. cholerae were isolated from 41 (3.8%) of total 1063 stool samples received during the study period. On serotyping, 100% (41) of these positive samples were identified as V. cholerae 01 ogawa serotype. 95% (39) cases were encountered during monsoon season. In the present study analysis of the antibiotic susceptibility data showed highest susceptibility to gentamicin, amikacin, tetracycline and chloramphenicol. There is gradual increase in resistance to ampicillin and high level of resistance was observed for furazolidone and trimethoprim-sulphamethoxazole during the same period. Conclusions: Our study reveals a significant increase in cases of Vibrio infection during monsoon season, with V. cholerae 01ogawa as the predominant serotype. The results also suggest that this organism has begun to develop resistance against ampicillin, trimethoprim-sulphamethoxazole and furazolidone drugs and therefore these should not be used in the first line treatment.
Keywords: Antibiotic resistance, cholera, diarrhea, India, vibrio cholera
|How to cite this article:|
Kumar A, Oberoi A. Vibrio isolates from cases of acute diarrhea and their antibiotic susceptibility pattern in a tertiary care hospital of Punjab. CHRISMED J Health Res 2014;1:254-7
|How to cite this URL:|
Kumar A, Oberoi A. Vibrio isolates from cases of acute diarrhea and their antibiotic susceptibility pattern in a tertiary care hospital of Punjab. CHRISMED J Health Res [serial online] 2014 [cited 2020 Dec 4];1:254-7. Available from: https://www.cjhr.org/text.asp?2014/1/4/254/142995
| Introduction|| |
Cholera is an acute type of diarrheal illness that affected millions of people around the world over the centuries. Vibrio cholera, the causative organism of cholera was first isolated by Robert Koch in 1883.  Of recent, V. cholerae 0139 has also been associated with severe forms of cholera.  In most parts of developing nations in the world Vibrio cholerae O1, the causative organism of epidemic cholera, continues to be a major health problem.  Especially during monsoon season, cholera outbreaks have been frequently reported from one or the other part of this country. However, occasional sporadic cases have also been reported, majorly due to lack of proper sanitation and impure water supply. , Therefore, public health strategy should ensure clean water supply, good sewage management and a clean environment.
The replacement of fluid and electrolytes plays a major role in treating cases of cholera however, treatment with antimicrobial agents has been proven effective in reducing intensity and duration of symptoms and also in preventing lethal complications.  Vibrio sp. were susceptible to many antimicrobials initially, but they have developed resistance to them over a period of time. Literature on the antibiotic susceptibility of cholera organisms from most developing countries is patchy. Worldwide, V. cholerae 01 strains resistant to tetracycline, trimethoprim-sulfamethoxazole and ampicillin are common. ,,,, In many of these studies, the main reasons for the rapid rise in antimicrobial resistance have been extensive antimicrobial prophylaxis, unauthorized dispensing and use of these agents in animal husbandry. , Resistance of V. cholerae to ceftriaxone has been reported from Argentina  and recently from Delhi, India.  Also, no consolidated data are available on the antimicrobial susceptibility patterns of organism concerned. The present study was conducted to determine the prevalence of serotypes of Vibrio isolates from cases of acute diarrhea and their antibiotic susceptibility pattern in a tertiary care hospital, North India.
| Materials and Methods|| |
This retrospective study was conducted over a period of one year from 1 st January 2013 to 31 th December 2013 in the Department of Microbiology, Christian Medical College and Hospital, Ludhiana, India, a tertiary care hospital. During this period, 1063 stool samples were collected from diarrheal/dysenteric patients admitted to the hospital. The patients included children, adults, and elderly people.
Fresh feces of patients were collected and transported immediately for culture. Stool samples received in the laboratory was proceeded to confirm the causative organism. The diagnosis was suggested by the clinical picture and confirmed by stool culture. Macroscopic examination of stool samples from the cases of acute watery diarrhea was done. Hanging drop preparation was made and observed for darting motility. The stool samples were inoculated onto MacConkey agar and Thiosulphate-citrate-bile salts-sucrose agar (TCBS) and Blood agar using standard microbiological techniques. Enrichment was done with alkaline peptone water, from which subculture was performed after six hours of incubation at 37°C onto MacConkey agar and Thiosulphate-citrate-bile salts-sucrose agar. Suspected colonies were identified as V. cholerae by the standard biochemical tests. ,, and Organisms biochemically resembling Vibrio were serologically confirmed by specific antisera (Denka Seiken, Japan). The Slide agglutination was performed with Vibrio cholera polyvalent 01, monospecific Ogawa and Inaba, and Vibrio cholera 0139 antisera. The susceptibility of all the isolated Vibrio species to different antibiotics were done by Kirby-Bauer's disk diffusion technique as per the Clinical and Laboratory Standards Institute (CLSI) guidelines. ,, The antibiotic disks tested were tetracycline (30 μg), furazolidone (100 μg), ampicillin (10 μg), trimethoprim-sulphamethoxazole (25 μg), chloramphenicol (30 μg), amikacin (10 μg), gentamicin (10 μg), ciprofloxacin (5 μg), norfloxacin (10 μg), and ceftriaxone (30 μg). Escherichia coli ATCC 25922 were used as the control organism.
| Results|| |
Out of total 1,063 stool samples received in the laboratory during the study period, Vibrio. Cholera was isolated in 41 stool samples, giving a positivity rate of 3.8%. On serotyping, 100% (41) of these positive samples were identified as V. cholerae 01 ogawa serotype. ninety-five percent (39) cases were encountered during monsoon season. These cases were usually documented in May to August of every year.
The prevalence of resistance pattern of the isolated V. cholerae 01 strains during the study period is as shown in the [Table 1]. Over 70% of Vibrio isolates were resistant to two or more drugs including ampicillin, trimethoprim-sulfamethoxazole and furazolidone. Resistance rate of ampicillin was 26.8%, trimethoprim-sulfamethoxazole 95.1%, and furazolidone 70.7% approximately. Two percent of Vibrio isolates were resistant to norfloxacin and ciprofloxacin, during the study period. No resistance was observed to tetracycline, Chloramphenicol, amikacin, gentamicin and third generation cephalosporins.
|Table 1: Percentage of Vibrio serogroups resistant to different antibiotics|
Click here to view
| Discussion|| |
In recent years, the emergence and the spread of resistance to antibiotics among Gram-negative organisms have been increasing rapidly. The epidemiological importance of preventing these drug resistant strains from spreading in the community has become a worldwide problem.  Particularly in developing countries, V. cholerae is a potent pathogen which can be transmitted easily in the community by the feco-oral route, in the existing sanitary systems. The gold standard of treatment consists of oral and intravenous rehydration therapy and only a small proportion with severe disease require antibiotic treatment.
Worldwide, there is great variation in the distribution of antibiotic resistant V. cholerae 01 strains. ,,, Our results suggest that the antimicrobial susceptibility patterns of V. cholerae 01 cannot be predicted easily. In India, like in many other developing countries, there is no control in the prescription of antimicrobial agents and that antibiotics are widely used in animal husbandry, which may account for the problem of increasing spread of antimicrobial resistance.
The prevalence of resistance pattern of the isolated V. cholera O1 strains during the study period is as shown in the [Table 1]. In our study, the isolates depicted highest susceptibility to tetracycline, Gentamicin, amikacin and Chloramphenicol. These findings were in concurrence to other studies in which the isolates were highly susceptible to aminoglycosides and tetracycline's. , . There is gradual increase in resistance to ampicillin and high level of resistance was observed for furazolidone and trimethoprim-sulphamethoxazole during the same period. A number of multidrug resistance strains are also emerging in clinical isolates of V. cholera 01; serotype Ogawa, including resistance to trimethoprim-sulphamethoxazole and ampicillin.  Resistance to fluoroquinolones has been reported in studies from India  , however most of the isolates were found to be susceptible to fluoroquinolones (Norfloxacin and Ciprofloxacin) in the present study.
The predominance of the Ogawa serotype (n = 41; 100%) in year 2013 suggests that it is currently the predominant circulating serotype in this part of the country. The seasonal association of cases observed. The clinical courses found to be more severe in children and the peak being during the monsoons, which declined remarkably with change of season. Most cases occurred in May to June, and the clustering of cases was observed till August.
| Conclusion|| |
Now the worldwide antimicrobial resistance has attained the importance of a public-health problem. Apart from the environmental stresses, which the pathogen is facing over the years, various factors are also responsible for increase in the magnitude of bacterial species resistant to multiple agents.
In developing countries, like India the main reason which is responsible for emergence of the expanding resistance patterns to the organism is the easy availability of over the counter drugs and wide spread institution of irrational chemoprophylaxis before the patient reaches a tertiary care hospital. , The emergence of drug resistant cholera may be due to overuse of these drugs as they are empirically used for many infections like diarrhea and urinary tract infection. This indiscriminate use of such antimicrobials will soon result in their becoming ineffective against this organism. With the emergence of high resistance to the empirically-given antibiotics, therapeutic options are limited. This problem becomes more dangerous and alarming when the issue of treating cholera in special groups, such as children, women in the antenatal period, and lactating mothers arises. The continuous surveillance of drug resistant strains is very important to know the changing antibiotic susceptibility pattern, as well as the cyclical change of the serogroups from time to time as the resistance pattern also changes with the change in the serogroups. Analysis and periodic reporting is important in proper therapy of cholera.
| References|| |
Sakazaki R. Bacteriology of vibrio and related organisms. In: Barua CD, Greenough W, editors. Plenum Medical Book Company; 1992. p. 37-54.
Albert MJ. Vibrio cholerae 0139 Bengal. J Clin Microbiol 1994;32:2345-9.
Cholera in 1997. Wkly Epidemiol Rec 1998;73:201-8.
Mhalu FS, Mmari PW, Ijumba J. A rapid emergence of El Tor Vibrio cholerae resistant to antimicrobial agent during first six months of fourth cholera epidemic in Tanzania. Lancet 1979;1:345-7.
Killewo JZ, Amsi DM, Mhalu FS. An investigation of a cholera epidemic in Butiama village of Mara Region, Tanzania. J Diarrhoeal Dis Res 1989;7:13-7.
Roychowdhury A, Pan A, Dutta D, Mukhopadhyay AK, Ramamurthy T, Nandy RK, et al.
Emergence of tetracycline-resistant Vibrio cholerae O1 serotype Inaba, in Kolkata, India. Jpn J Infect Dis 2008;61:128-9.
Finch MJ, Morris JG Jr, Kaviti J, Kagwanja W, Levine MM. Epidemiology of antimicrobial resistant cholera in Kenya and East Africa. Am J Trop Med Hyg 1988;39:484-90.
Mohamedali HF, Mhalu FS, Lyamuya EF. Susceptibility of Salmonella, Shigella and V. cholerae 01 to antimicrobial agents at Muhimbili Medical Centre, Dares Salaam during 1990-91. Tanzania Med J 1992:14-7.
Olukoya DK, Ogunjimi AA, Abaelu AM. Plasmid profiles and antimicrobial susceptibility patterns of Vibrio cholerae 01 strain isolated during a recent outbreak in Nigeria. J Diarrhoeal Dis Res 1995;13:118-21.
Materu SF, Lema OE, Mukunza HM, Adhiambo CG, Carter JY. Antibiotic resistance pattern of Vibrio cholerae and Shigella causing diarrhoea outbreaks in the Eastern African region: 1994-1996. East Afr Med J 1997;74:193-7.
Dalsgaard A, Forslund A, Tam NV, Vinh DX, Cam PD. Cholera in Vietnam: Changes in genotypes and emergence of class 1 integrons containing aminoglycosides resistance gene cassettes in Vibrio cholerae 01 strains isolated from 1979 to 1996. J Clin Microbiol 1999;37:734-41.
Thornsbery C. Trends in antimicrobial resistance among today's bacterial pathogens. Pharmacotherapy 1995;15:3-8S.
Resistance to antimicrobial agents. Wkly Epidemiol Rec 1997;45:333-6.
Petroni A, Corso A, Melano R, Cacace ML, Bru AM, Rossi A, et al
. Plasmidic extended-spectrum beta- lactamases in Vibrio cholerae O1 El Tor isolates in Argentina. Antimicrob Agents Chemother 2002;46:1462-8.
Sharma NC, Mandal PK, Dhillon R, Jain M. Changing profile of Vibrio cholerae O1, O139 in Delhi and its periphery (2003-2005). Indian J Med Res 2007;125:633-40.
Centers for Disease Control and Prevention. Laboratory methods for the diagnosis of epidemic dysentery and cholera. Atlanta, GA: Centers for Disease Control and Prevention; 1999. p. 41-54. (WHO/CDS/CSR/EDC/99.8).
Old DC. Vibrio, Aeromonas, Plesiomonas, Campylo- bacter, Arcobacter, Helicobacter, Wolinella. In: Collee JG, Fraser BP, Marmion AG, Simmons A, editors. Mackie and McCartney Practical medical microbiology. 14 th
ed. Edinburgh: Churchill Livingstone; 1999. p. 425-48.
Bradford KA, Cheryl BA, Joy WG. Isolation and identification of Vibrio cholerae O1 from fecal specimens. In: Wachsmuth IK, Blake PA, Olsvik O, editors. Vibrio cholerae and cholera: Molecular to global perspectives. Washington: American Society for Microbiology Press; 1994. p. 3-25.
Bauer AW, Kirby WM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardised single disk method. Am J Clin Pathol 1966;45:493-6.
Centers for Disease Control and Prevention. Manual for the laboratory detection of antimicrobial resist- ance among acquired bacterial pathogens of public health concern in the developing world (draft). At- lanta: Centers for Disease Control and Prevention; 2001. p. 55-64. (CDC/WHO/USAID).
Clinical and Laboratory Standards Institute. Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolated or fastidious bacteria; approved guideline. 2 nd
ed. Wayne: Clinical and Laboratory Standards Institute; 2010. (M45A2).
Taneja N, Samanta P, Mishra A, Sharma M. Emergence of tetracycline resistance in Vibrio cholerae O1 biotype El Tor serotype Ogawa from north India. Indian J Pathol Microbiol 2010;53:865-6.
Hofer E, Quintaes B, dos Reis E, Rodrigues D, Seki L, Feitosa I, et al
. The emergence of multiple antimicrobial resistance isolated from gastro-enteritis patients in Ceara, Brazil. Rev Soc Med Trop 1999;32:151-6.
Taneja N, Kaur J, Sharma K, Singh M, Kalra JK, Sharma NM, et al
. A recent outbreak of cholera due to Vibrio cholerae O1 Ogawa in and around Chandigarh, North India. Indian J Med Res 2003;117:243-6.
Kutar BM, Rajpara N, Upadhyay H, Ramamurthy T, Bhardwaj AK. Clinical isolates of Vibrio cholerae O1 El Tor Ogawa of 2009 from Kolkata, India: Preponderance of SXT element and presence of Haitian ctxB variant. PLoS One 2013;8:e56477.
Mukhopadhyay AK, Basu I, Bhattacharya SK, Bhattacharya MK, Nair GB. Emergence of fluoroquinolone resistance in strains of Vibrio cholerae isolated from hospitalized patients with acute diarrhea in Calcutta, India. Antimicrob Agents Chemother 1998;42:206-7.
Jesudason MV, Saaya R. Resistance of vibrio cholerae O1 to nalidixic acid. Indian J Med Res 1997;105:153-4.
O'Brien TF. The global epidemic nature of antimicrobial resistance and the need to monitor and manage it locally. Clin Infect Dis 1997;24 (Suppl 1):S2-8.