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 ~  Abstract
 ~  Materials and Me...
 ~  Results
 ~  Discussion
 ~  Acknowledgements
 ~  References

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Year : 2003  |  Volume : 21  |  Issue : 3  |  Page : 186-188
 

Y.enterocolitica gastroenteritis - A prospective study


Department of Microbiology, Christian Medical College, Ludhiana -141 008, Punjab, India

Correspondence Address:
Department of Microbiology, Christian Medical College, Ludhiana -141 008, Punjab, India

 ~ Abstract 

Two thousand faecal samples were collected from diarrhoeal patients of all age groups and processed by using MacConkey agar and Yersinia selective agar [Cefsulodin-Irgasan-Novobiocin (CIN) media]. One hundred and seventy faecal samples from non-diarrhoeal (healthy controls) were also processed. Samples were subjected to cold enrichment before processing. Three hundred and seventy-four (18.7%) faecal samples were positive for bacterial pathogens, of which 41 (2.05%) had Y.enterocolitica infection. Y.enterocolitica was more common in patients above 16 years of age 23/41 (56%). Higher rates of isolation were recorded during winter months. Of the typable isolates 21.9% belonged to 0:3 serotype followed by 01, 2a, 3 (17%) serotype. Twelve (29.2%) isolates were untypable. 51.2% isolates of Y.enterocolitica belonged to biotype IV whereas 24.4%, 19.5%, 4.9% belonged to type I, II and III respectively. Majority of the isolates were found sensitive to chloramphenicol, gentamicin, amikacin, ciprofloxacin, ampicillin, netromycin, cefoperazone, tobramycin and pefloxacin. All isolates were resistant to ofloxacin.

How to cite this article:
Lal M, Kaur H, Gupta L K. Y.enterocolitica gastroenteritis - A prospective study. Indian J Med Microbiol 2003;21:186-8


How to cite this URL:
Lal M, Kaur H, Gupta L K. Y.enterocolitica gastroenteritis - A prospective study. Indian J Med Microbiol [serial online] 2003 [cited 2020 Oct 20];21:186-8. Available from: https://www.ijmm.org/text.asp?2003/21/3/186/8013


Y.enterocolitica, a gram negative aerobic bacterium is a member of the family Enterobacteriacae. It is rapidly emerging worldwide as an enteric pathogen associated with a wide spectrum of clinical and post immunological manifestations including acute gastroenteritis, mesenteric adenitis, septicaemia, arthritis and erythema nodosum, predominantly affecting young children.[1] Y.enterocolitica has become a major cause of diarrhoea in most of the industrialized world. Y.enterocolitica being psychrophilic is very tolerant to low temperature and can continue to grow at temperature as low as 1C. Y.enterocolitica has been isolated from vacuum packed meats and can survive for extended periods in frozen food, even with repeated freezing and thawing. Y.enterocolitica was first isolated in Asia from Japan.[2]
Sporadic cases of human gastroenteritis due to Y.enterocolitica have been recognized with increasing frequency since then. Y.enterocolitica rivals  Salmonella More Details and even surpasses shigella in causing acute enteritis. In several tropical countries such as Iran, Israel, South Africa and Brazil, Y.enterocolitica has been observed to be a significant cause of human infections.[3] In India, very scanty literature is available regarding this organism. This study was undertaken to assess the incidence of diarrhoea caused by Y.enterocolitica in the northern part of the country.

[TAG:2]Materials and Methods[/TAG:2]
A total of 2000 faecal samples from patients of all age groups (both inpatients and outpatients) suffering from gastroentritis attending Christian Medical College and Hospital, Ludhiana, Punjab and 170 faecal samples from non-diarrhoeal (healthy controls) subjects were collected and studied from December, 1996 to December, 1999. Christian Medical College (CMC) and Hospital is a tertiary care hospital and patients seen are from Punjab and adjacent states.
Cold enrichment and Isolation of Y.enterocolitica
About 1-2 grams of faecal sample was added to a tube containing 5 mL of phosphate buffered saline (pH 7.2) for cold enrichment at 40C for two weeks.[4] Subcultures were done on 7th and 14th day on MacConkey agar and  Yersinia More Details selective agar. Plates were incubated for 24-48 hours.
Identification, antibiotic susceptibility and typing of Y.enterocolitica
Each isolate of suspected Y.enterocolitica was subcultured on a set of MacConkey agar and blood agar. One set of each was incubated overnight at 250C and 370C respectively. It was also subjected to Gram staining, motility by hanging drop and soft agar method incubating the culture media at 250C as well as 370C. In addition, the biochemical tests were put up for identification.[5] Isolates were biotyped on the basis of results of various biochemical reactions - indole production, sucrose utilization, trehalose utilization, xylose utilization, lipase activity and nitrate reduction. Nine of the 41 isolates were serotyped at Central Public Health Laboratory, Colindale, London and 32 isolates were serotyped by slide agglutination using antisera from Siken Denka Company, Tokyo, Japan. Sensitivity to the various antibiotics was determined by Stoke's disc diffusion technique.

 ~ Results Top

Out of 2000 faecal samples processed only 41(2.05%) were found to be positive for Y.enterocolitica. Y.enterocolitica was not isolated from any of the 170 healthy controls. No major difference was observed in male-female ratio (22:19). Y. enterocolitica was isolated from 18 children (upto 15 yrs age) and 23 adults. In this study the Y.enterocolitica diarrhoea was more common in patients above 16 years of age (1.15%) followed by 1-5 years (0.4%) age group. Infants constituted the third largest group (0.35%). All patients presented with loose stools, 31.7% of the patients presented with loose stools and vomiting, 29.3% with abdominal pain, 24.3% with fever and 12.1% of the patients were found to be severely dehydrated.
On MacConkey agar, the colonies of Y.enterocolitica were minute, non-lactose fermenting (NLF), circular, smooth, translucent, glistening and convex with an entire edge. On blood agar, colonies appeared like colonies of Enterococcus. On  Yersinia More Details selective agar the colonies appeared like bull's eye. They were all gram negative rods and motile at 250C and non-motile at 370C. All the 41 isolates were positive for catalase, methyl red reaction, reduced nitrate to nitrite, hydrolysed urea and all the isolates were negative for oxidase, phenyl pyruvic acid, H2S production and citrate was not utilised by any isolate. All isolates fermented glucose, sucrose, mannitol, maltose, sorbitol, arabinose and trehalose. The isolates did not ferment lactose, xylose, raffinose and rhamnose.
Antibiotic sensitivity
The drug sensitivity revealed that all the isolates were sensitive to chloramphenicol, gentamicin, amikacin, ciprofloxacin, amoxycillin, netromycin, cefoperazone, tobramycin and pefloxacin. Sensitivity to tetracycline and ceftriaxone was seen in 92.8% isolates where as sensitivity to cotrimoxazole, cephalexin and ampicillin was seen in 78.5, 57.1, 7.2% isolates respectively. All isolates were resistant to ofloxacin.
Biotypes and Serotypes
Biotyping revealed that the most common biotype was type-IV (51.2%) and 24.4, 19.5, 4.9% respectively belonged to type I, II and III [Table - 1].
Nine out of 41 (21.9%) of the isolates belonged to serotype 0:3, 7/41 (17%) were of serotype 01,2a, 3, 12/41 (29.2%) isolates were un-typable and the rest belonged to other serotypes of 0:1,0:2, 035 and O19,8 [Table - 1].

 ~ Discussion Top

Y.enterocolitica was considered a rare microorganism for a long time, but during the last two decades it has been isolated all over the world from animals, raw food materials, environment, water and human beings.[6] There are only few reports of Y.enterocolitica isolations from human beings in India despite frequent description of human infections with Y.enterocolitica in Europe, North America, Brazil, some African countries, Japan and Iran. This may be attributed to the low incidence of this organism in human infections, as has been revealed in this study, combined with a lack of awareness about the occurrence of this organism. The rate of isolation in this study is 2.05% which probably indicates rarity of this organism in this country. Onyemelukwe isolated 9(1.4%) Y.enterocolitica from faecal samples of children between the age group of 0-12 years in Nigeria.[7] The rate of isolation of this organism from diarrhoeal stools from several other countries has been reported to be 2 to 5%.
The age distribution pattern of Y.enterocolitica diarrhoea in this study was found to be similar to that reported from other countries except that the highest isolation was found in patients above 16 years of age. This correlates well with a study by Engelaer and Konstainje who found 109 (42.1%) patients above 25 years of age infected with Y.enterocolitica.[8] Higher rates of isolation were recorded in winter months (November to January). This correlates with results by Lee et al [9] who reported maximum number of isolates during November to January. The drug sensitivity pattern correlates well with report from Taipei.[10] Valera found ceftriaxone, ciprofloxacin and amikacin to be effective for the treatment of Y.enterocolitica.[11] Similarly, all isolates were sensitive to these drugs in this study.
Nine of the 30 typable isolates belonged to serotype 0:3. In Europe, Canada, Latin American, South Africa and Japan, serotype 0:3 is most common. Seven of our isolates belonged to 01, 2a, 3, which is comparable to a study by Ram et al[12] who isolated 7 Y.enterocolitica isolates from 235 faecal samples and all their isolates belonged to the serogroup of 01,2a,3. In this study, there were five isolates of serotype 0:19,8, four of 0:9 and one each of 0:2, 03:5, 0:1. For some reason, not yet understood, 0:3, 0:9 are rare in United States whereas serotype 0:8 predominates. Serotype 0:3 has been reported from Malaysia. The prevalence of different Y.enterocolitica serotypes is quite characteristic in its geographical distribution.

 ~ Acknowledgements Top

The authors are grateful to Dr. Betty C Hobbs, Central Public Health Laboratory, Colindale, London for serotyping of the isolates, Dr.George Kaupperad, Public Health Laboratory, Oslo, Norway for the supply of Laboratory reagents, and ESP Germany for the financial assistance. 

 ~ References Top

1.Bottone EJ, Robin T. Prevalence of unique Y.enterocolitica in the area of Mount Sinai Hospital, New York, N.Y. Contrib Microbiol Immunol 1979;5:95-98.  Back to cited text no. 1    
2.Zen-Yoji H, Maruyamma T. The first successful isolation and identification of Y.enterocolitica from human cases in Japan. Jpn J Microbiol 1972;16:493-495.  Back to cited text no. 2    
3.Pramanik AK, Bhattacharya HM, Chatterjee A, Sen Gupta DN. Isolation of Y.enterocolitica from human and animals from the same family. Indian J Animal Health 1980;19:79-83.  Back to cited text no. 3    
4.Peterson JS, Cook RA. Method for recovery of Pasteurella pseudotuberculosis from faeces. J Pathol Bacteriol 1963;85:241-243.  Back to cited text no. 4    
5.Cowan ST. Manual for the identification of medical bacteria, 3rd ed. (Cambridge University Press, Cambridge) 1974;97-164.  Back to cited text no. 5    
6.Kapperud G. Yersinia and Yersinia like microbes isolated from mammals and water in Norway and Denmark. Acta Pathol Microbiol Scand 1977;85:129-134.  Back to cited text no. 6    
7.Onyemelukwe NF. Yersinia enterocolitica as an aetiological agent of childhood diarrhoea in Enugu Nigeria. Cent Afr J Med1993;39(9):192-195.  Back to cited text no. 7    
8.Stolk-Engelaar VM, Hoogkamp-Korstonje JA. Clinical presentation and diagnosis of gastroenteritis infection by Y.enterocolitica in 261 Dutch patients. Scand J Infect Dis 1996;28(6):571-574.  Back to cited text no. 8    
9.Lee LA, Taylor J, Carter GP, Quinn B, Farmer JJ 3rd, Tauxe RV. Yersinia enterocolitica O:3: An emerging cause of pediatric gastroenteritis in the United States. The Yersinia enterocolitica collaborative study group. J Infect Dis 1991;163:660-663.  Back to cited text no. 9    
10.Ding MJ, Chiang HY, Lin CL, Williams RP. Isolation and characterization of Y.enterocolitica in Taipei. Chinese J Microbiol Immunol 1986;19(4):295-301.  Back to cited text no. 10    
11.Valera MJ, Gonzalez TC, Moreno E, Ruiz-Bravo. A comparison of ceftriaxone, amikacin and ciprofloxacin in treatment of experimental Y.enterocolitica O:9 infection in mice. Antimicrob Agents Chemother 1998;11(42): 3009-3011.  Back to cited text no. 11    
12.Ram S, Khurana S, Singh R, Sharma S, Vadehra DV. Y.enterocolitica diarrhoea in north India. Indian J Med Res 1987;86:9-13.  Back to cited text no. 12    
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