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Open Access
Copyright: The authors. This article is an open access
article licensed under the terms of the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/2.0) which permits unrestricted use,
distribution and reproduction in any medium, provided the work is properly
cited.
Research
(Published
online : 07-07-2013)
15. Isolation of pathogenic Escherichia coli from stool samples of diarrhoeal
patients with history of raw milk consumption - Paresh K. Virpari, J.
B. Nayak, H. C. Thaker and M. N. Brahmbhatt
Vet World. 2013; 6(9):
659-663
doi:
10.14202/vetworld.2013.659-663
Abstract
Aim: To detect the occurrence
of pathogenic Escherichia coli from stool samples of diarrhoeal
patients with history of raw milk consumption and to determine the
public health significance of isolates, especially their role in
causing human diseases.
Materials and Methods: A total of 100 stool samples from
diarrhoeal patients, with history of raw milk consumption were
collected from primary health centres in and around Anand city,
under aseptic conditions and a total of 50 raw milk samples were
collected from milk vendors, retail shops located in Anand city in
sterilized sample bottles. MacConkey broth was used for the
enrichment of all the samples and inoculation was done on
MacConkey agar and EMB agar was used as the selective media. This
was followed by the confirmation of isolates using biochemical
tests. For the serotyping, E. coli isolates were sent to the
National Salmonella and Escherichia Centre, Central Research
Institute (CRI), Kasauli, Himachal Pradesh. Detection of virulence
genes was performed using PCR technique.
Results: During the present investigation, 26 (52%) E. coli
isolates from 50 milk samples and 59 (59%) E. coli isolates from
100 stool samples were recovered. Out of 85 E. coli isolates sent
for serotyping, 74 isolates could be typed which were further
distributed into 13 different serogroups O2, O4, O8, O17, O22,
O25, O29, O36, O45, O60, O90, O116 and O172, whereas 8 isolates
were found untypable and 3 isolates were reported rough isolates.
Of the 59 E. coli isolates from stool samples of diarrhoeal
patients tested, 15 isolates (25.42%) were reported to be positive
for stx genes, among that 6 (10.16%) were positive for stx1 gene,
9 (15.25%) isolates were positive for stx2 gene, while 3 isolates
(5.08%) were positive for eaeA gene. In this study, 21 E. coli
isolates were found to be Shiga toxin producing E. coli (STEC)
while none of the isolates were positive for the serotype O157.
Conclusions: Our present findings indicate that raw milk
may act as a source of pathogenic E. coli and it may be
responsible for the occurrence of diarrhoea and various other
health-related complications in humans. We therefore recommend
proper managemental practices and effective control measures for
improved hygiene and sanitation.
Keywords: diarrhoea, E. coli, milk, serotype, shiga toxin
producing E. coli , stools
References
1. Tchaptchet, S. and Hansen, J. (2011) The Yin and Yang of
host-commensal mutualism. Gut Microbes.2: 347–352.
http://dx.doi.org/10.4161/gmic.19089
PMid:22198619 |
|
|
|
2. Farrokh, C., Kieran, J., Auvray, F., Glass, K., Oppegaard,
H., Raynaud, S., Thevenot, D., Condron, R., De Reu, K.,
Govaris, A., Heggum, K., Heyndrickx, M., Hummerjohann, J.,
Lindsay, D., Miszczycha, S., Moussiegt, S., Verstraete, K. and
Cerf, O. (2012) Review of Shiga-toxin-producing Escherichia
coli (STEC) and their significance in dairy production. Int.
J. Food Microbiol. http://dx.doi.org/
10.1016/j.ijfoodmicro.2012.08.008,10/04/2013. |
|
|
3. Karmali, M. A., Gannon, V. and Sargeant, J. M. (2010)
VerocytotoxinproducingEscherichia coli (VTEC). Vet. Microbiol.
140: 360–370.
http://dx.doi.org/10.1016/j.vetmic.2009.04.011
PMid:19410388 |
|
|
4. Caprioli, A., Morbito, S., Brugere, H. and Oswald, E.
(2005) EnterohaemorrhagicEscherichia coli: emerging issue on
virulence and mode of transmission. Vet. Res. 36: 289–311.
http://dx.doi.org/10.1051/vetres:2005002
PMid:15845227 |
|
|
5. Lorusso, V., Dambrosio, A., Quaglia, N. C., Parisi, A., La
Salandra, G., Lucifora, G., Mula, G., Virgilio, S., Carosielli,
L., Rella, A., Dario, M. and Normanno, G. (2009) Verocytotoxin-producing
Escherichia coli O26 in raw water buffalo (Bubalusbubalis)
milk products in Italy. J. Food Prot.72(8): 1705-1708.
PMid:19722404 |
|
|
6. Buvens, G., Posse, B., De Schrijver, K., De Zutter, L.,
Lauwers, S. and Pierard, D. (2011) Virulence profiling and
quantification of verocytotoxin-producing Escherichia coli
O145:H28 and O26:H11 isolated during an ice cream-related
haemolytic uremic syndrome outbreak. Foodborne Pathogens and
Disease. 8: 421–426.
http://dx.doi.org/10.1089/fpd.2010.0693
PMid:21166561 |
|
|
7. Kumar, R., Surendran, P. K. and Thampuran, N. (2008)
Evaluation of culture, ELISA and PCR assays for the detection
of Salmonella in seafood. Letters in Applied Microbiol. 46(2):
221-226.
http://dx.doi.org/10.1111/j.1472-765X.2007.02286.x
PMid:18028326 |
|
|
|
8. Edwards, P. R. and Ewing, W. H. (1972)Identification of
Enterobacteriaceae. Emerg. Infect. Dis. 12:154–159. |
|
|
9. Beverley, C., Millar, X. J., John, E. M. and John A. P. E.
(2000) A simple and sensitive method to extract bacterial,
yeast and fungal DNA from blood culture material. Journal of
Microbiological Methods. 42: 139-147.
http://dx.doi.org/10.1016/S0167-7012(00)00174-3 |
|
|
10. Osman, K. M., Mustafa, A. M., Elhariri, M. and AbdElhamed,
G. S. (2013) The Distribution of Escherichia coli serovars,
virulence genes, gene association and combinations and
virulence genes encoding serotypes in pathogenic E. coli
recovered from diarrhoeic calves, sheep and goat. Transbound
Emerg. Dis. 60(1):69-78.
http://dx.doi.org/10.1111/j.1865-1682.2012.01319.x
PMid:22435607 |
|
|
11. Dhanashree, B. and Mallya, P. S. (2008) Detection of shiga-
toxigenic Escherichia coli (STEC) in diarrhoeagenic stool &
meat samples in Mangalore, India. Indian J. Med. Res. 128:
271-277.
PMid:19052337 |
|
|
12. Beutin, L., Montenegro, M. A., Orskov, I., Orskov, F.,
Prada, J., Zimmermann, S. and Stephan, R. (1989)Close
association of verotoxin (shiga-like toxin) production with
enterohemo- lysin production in strains of Escherichia coli.
J. Clin. Microbiol. 27: 2559–2564.
PMid:2681256 PMCid:PMC267076 |
|
|
13. Rivas, M., Miliwebsky, E., Chinen, I., Roldan, C. D.,
Balbi, L., Garcia, B., Fiorilli, G., Sosa-Estani, S., Kincaid,
J., Rangel, J. and Griffin, P. M. (2006) Characterization and
epidemiologic subtyping of Shiga toxin-producing Escherichia
coli strains isolated from hemolytic uremic syndrome and
diarrhea cases in Argentina. Foodborne Pathog. Dis. 3: 88-96.
http://dx.doi.org/10.1089/fpd.2006.3.88
PMid:16602984 |
|
|
|
14. Kausar, Y., Chunchanur, S. K., Nadagir, S. D., Halesh, L.
H. and Chandrasekhar, M. R. (2009) Virulence factors,
Serotypes and Antimicrobial Suspectibility Pattern of
Escherichia coli in Urinary Tract Infections. Al Ameen J. Med.
Sci. 2(1):47-51. |
|
|
|
15. Karama, M., Johnson, R. P., Holtslander, R., McEwen, S. A.
and Gyles, C. L. (2008) Prevalence and characterization of
verotoxin-producing Escherichia coli (VTEC) in cattle from an
Ontario abattoir. The Canadian J. Vet. Res. 72: 298-302. |
|
|
|
16. Nishikawa, Y., Zhou, Z., Hase, A., Ogasawara, J., Kitase,
T., Abe, N., Nakamura, H., Wada, T.,Ishii, E. and Haruki, K.
(2002) Diarrheagenic Escherichia coli isolated from stools of
sporadic cases of diarrheal illness in Osaka city, Japan
between 1997 and 2000: Prevalence of Enteroaggregative E. coli
heat stable enterotoxin 1 gene- possessing E. coli. Jpn. J.
Infect. Dis. 55:182-190. |
|
|
17. Brooks, J. T., Sowers, E. G., Wells, J. G., Greene, K. D.,
Griffin, P. M., Hoekstra, R. M. and Strockbine, N. A. (2005)
Non-O157 Shiga toxin-producing Escherichia coli infections in
the United States, 1983–2002. J. Infect. Dis. 192: 1422-1429.
http://dx.doi.org/10.1086/466536
PMid:16170761 |
|
|
18. Arthur, T. M., Barkocy-Gallagher, G. A.,
Rivera-Betancourt, M. and Koohmaraie, M. (2002) Prevalence and
characterization of non-O157 shiga toxin-producing Escherichia
coli on carcasses in commercial beef cattle processing plants.
App. Environ. Microbiol. 68(10): 4847- 4852.
http://dx.doi.org/10.1128/AEM.68.10.4847-4852.2002
PMid:12324330 PMCid:PMC126400 |
|
|
19. Leedom, J. M. (2006) Milk of nonhuman origin and
infectious diseases in humans. Clin. Infect. Dis. 43:610–615.
http://dx.doi.org/10.1086/507035
PMid:16886155 |
|
|
20. Guh, A., Phan, Q., Randall, N., Purviance, K., Milardo,
E., Kinney, S., Mshar, P., Kasacek, W. and Cartter, M. (2010)
Outbreak of Escherichia coli O157 Associated with Raw Milk,
Connecticut, 2008. Clin. Infect. Dis. 51(12): 1411-1417.
http://dx.doi.org/10.1086/657304
PMid:21058911 |
|
|
21. LeJeune J. T. andRajala-Schultz P. J. (2009) Food safety:
unpasteurized milk: a continued public health threat. Clin
Infect Dis. 48:93–100.
http://dx.doi.org/10.1086/595007
PMid:19053805 |
|
|
22. Solomakos, N., Govaris, A., Apostolos, S., Spyros, A.,
Angeliki, P., Burriel, R., Spyridon, K., Demetrios, K. and
Papageorgiou, K. (2009) Occurrence, virulence genes and
antibiotic resistance of Escherichia coli O157 isolated from
raw bovine, caprine and ovine milk in Greece. Food Microbiol.
26(8): 865–871.
http://dx.doi.org/10.1016/j.fm.2009.06.002
PMid:19835773 |
|
|
23. Stephan, R., Schumacher, S., Corti, S., Krause, G.,
Danuser, J. and Beutin, L. (2008) Prevalence and
characteristics of Shiga toxin-producing Escherichia coli in
Swiss raw milk cheeses collected at producer level. J. Dairy
Sci. 91(7): 2561- 2565.
http://dx.doi.org/10.3168/jds.2008-1055
PMid:18565913 |
|
|
24. Madic, J.,Vingadassalon, N., de Garam,C. P., Marault,
M.,Scheutz, F., Brugere, H. Jamet, E. and Auvray, F. (2011)
Detection of Shiga toxin-producing Escherichia coli serotypes
O26:H11, O103:H2, O111:H8, O145:H28, and O157:H7 in raw-milk
cheeses by using multiplex real-time PCR. Applied and
Environmental Microbiology 77(6): 2035–2041.
http://dx.doi.org/10.1128/AEM.02089-10
PMid:21239543 PMCid:PMC3067316 |
|
|
