www.wjpr.net Vol 3, Issue 4, 2014. 529 OCCURRENCE OF YERSINIA ENTEROCOLITICA IN DRINKING WATER IN THE ABSENCE OF INDICATOR ORGANISM Sahota P*, Sharma N, Kirandip and Pandove G Department of Microbiology, Punjab Agricultural University, Ludhiana, Punjab, India. ABSTRACT The drinking water quality with respect to bacteriological and physicochemical examination was done for 238 drinking water samples from different water utilities (municipal corporation supply taps, submersible pumps, filter samples, water tanks, storage tanks, water coolers, etc.) of urban areas of Ludhiana, Punjab, India. Standard methods were used for the analysis of pH, TDS, while E. coli, Yersinia spp. determination was carried out using BWTK and Multiple Tube Technique. Yersinia spp. was detected in 164 (68.91%) water samples. Statistical correlation was examined between the bacteriological and physicochemical parameters and found to be independent from each other. Difference between mean in each case, high standard deviation indicate that the distribution is widely off normal and exhibit an asymmetric distribution. The occurrence of E. coli was also not found to be associated with the presence or absence of Yersinia enterocolitica. So, E. coli was not found to be a suitable indicator for Yersinia spp. The findings of the current study reports the occurrence of Y. enterocolitica in drinking water and suggesting that there is need for further surveillance studies to understand the global epidemiology of emerging pathogens. Keywords: BWTK, correlation, indicator orgainsm, urban areas, drinking water, Yersinia enterocolitica. INTRODUCTION Safe water supplies and environmental sanitation are vital for protecting the environment, improving health and alleviating property. Deterioration in water quality below the established standards is mainly due to human activities and untreated or inadequately treated wastes discharge in the environment. The World Health Organisation (WHO) estimated that Article Received on 01 April 2014, Revised on 22 April 2014, Accepted on 15 May 2014 *Correspondence for Author Dr.(Mrs.) Param Pal Sahota, Sr.Microbiologist, Deptt. of Microbiology, PAU,Ludhiana- 141004, Punjab. World Journal of Pharmaceutical ReseaRch SJIF Impact Factor 5.045 Volume 3, Issue 4, 529-542. Research Articl ISSN 2277 – 7105
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www.wjpr.net Vol 3, Issue 4, 2014.
529
Sahota et al. World Journal of Pharmaceutical Research
OCCURRENCE OF YERSINIA ENTEROCOLITICA IN DRINKING
WATER IN THE ABSENCE OF INDICATOR ORGANISM
Sahota P*, Sharma N, Kirandip and Pandove G
Department of Microbiology, Punjab Agricultural University, Ludhiana, Punjab,
India.
ABSTRACT
The drinking water quality with respect to bacteriological and
physicochemical examination was done for 238 drinking water
samples from different water utilities (municipal corporation supply
taps, submersible pumps, filter samples, water tanks, storage tanks,
water coolers, etc.) of urban areas of Ludhiana, Punjab, India. Standard
methods were used for the analysis of pH, TDS, while E. coli, Yersinia
spp. determination was carried out using BWTK and Multiple Tube
Technique. Yersinia spp. was detected in 164 (68.91%) water samples.
Statistical correlation was examined between the bacteriological and
physicochemical parameters and found to be independent from each other. Difference
between mean in each case, high standard deviation indicate that the distribution is widely off
normal and exhibit an asymmetric distribution. The occurrence of E. coli was also not found
to be associated with the presence or absence of Yersinia enterocolitica. So, E. coli was not
found to be a suitable indicator for Yersinia spp. The findings of the current study reports the
occurrence of Y. enterocolitica in drinking water and suggesting that there is need for further
surveillance studies to understand the global epidemiology of emerging pathogens.
Table 2: Pearson value of correlation (r), along with levels of statistical significance
between various parameters
Parameters MPN P value
pH 0.280159 <0.0001*
TDS -0.09686 0.1362
* Test of significance at 1% level (P=0.01)
Table 3 Morphological and cultural characteristics of Y. enterocolitica isolates from
drinking water samples.
Morphological and cultural tests
Observations
Gram’s reaction Gram negative
Motility At 25˚C At 37˚C
Motile Non-motile
Shape Rods
CIN (Yersinia Selective Medium)
Bull’s eye morphology with deep red centers and translucent periphery
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Sahota et al. World Journal of Pharmaceutical Research
HiChrome UTI agar circular, smooth, convex colonies, with glistening surface
Trypticase soya agar circular, smooth, convex colonies and were easily emulsifiable
Nutrient agar circular, smooth, convex colonies, with glistening surface, 1-2 mm in diameter, entire or slightly crenated edges and were easily emulsifiable
Mac Conkey’s lactose agar At 25˚C At 37˚C
Small colourless, raised convex colonies having entire margin Large colourless, raised convex colonies having entire margin
Nutrient broth Yersinia enterocolitica produced uniform turbidity with some green colouration by some strains
Bacteriological Water Testing Kit (BWTK)
Forms superficial layer on the kit
Table 4 Comparison of fermentation of various sugars by isolates of Y. Enterocolitica.
Sugars utilization No. of isolates Y. enterocolitica (%)
Sahota et al. World Journal of Pharmaceutical Research
Table 5 Biochemical characteristics of Y. Enterocolitica.
Biochemical Characteristics No. of isolates Y. enterocolitica (%)
1. Urease test + (100%) 2. Indole test + (83%) 3. Citrate utilization - (100%) 4. Triple Sugar Iron (TSI) test - (100%) 5. Esculin hydrolysis at (28°C) At 37°C
- (100%) - (100%)
6. Methyl Red (MR) test + (100%) 7. Voges-Proskauer’s (VP) test - (100%) 8. Phenylalanine deaminase agar
test - (100%)
9. Nitrate reduction test + (100%) 10. Oxidase test - (100%) 11. Orthonitrophenyl-β-
galactosidase (ONPG) + (100%)
12. Lysine decarboxylase - (100%) 13. Ornithine decarboxylase + (100%) 14. Arginine dihydrolase - (100%) 15. Gelatinase production - (100%) 16. H2S production - (100%) 17. Catalase test + (100%)
Table 6 Relationship of occurrence of Yersinia spp. with E. coli (chi-square test using
Yates correction).
E. coli
Yersinia spp.
Absent Present
Absent 41 33
Present 87 77 χ2 value (Calculated value) = 0.039 (at 1 degree of freedom)
Table value (P value) = 0.4219 (P<0.01)
So, there is no association between the occurrence of Yersinia spp. and E. coli.
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Sahota et al. World Journal of Pharmaceutical Research
Table 7 Pathogenicity tests
Pathogenicity tests Observations
Congo red dye uptake Red coloured colonies
Crystal violet binding Violet coloured colonies
Autoagglutination test An irregularly edged layer of agglutinated
bacteria which formed flocculate covering at the
bottom of the tube with clear supernatant fluid
Figure 1: Percentage of occurrence of Yersinia enterocolitica in various months
CONCLUSION
Few developed countries have included emerging pathogens in their standards for drinking
water and hence require occurrence of emerging pathogens in water and foods to be reported
to authorities for the safety of health but these pathogens are not included in water and food
standards of any developing country. The water samples of study area were highly polluted
bacteriologically. These results also demonstrated the presence of widespread, biochemically
and pathogenically characterized, waterborne Y.enterocolitica in urban areas of Punjab. The
detection of Y.enterocolitica in drinking water sources studied represent a public health
concern, and must be taken into account for assessing the quality of drinking water. We
would like to recommend the following important points: proper sanitary survey, design and
implementation of water and/ or sanitation projects; regular disinfection, maintenances and
supervisions of water sources and regular bacteriological assessment of all water sources for
drinking should be planned and conducted. Systems using surface water sources are required
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to disinfect to ensure that all bacterial contamination (such as E. coli and Yersinia spp.) is
inactivated.
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