Detection of E.Coli Strains Isolated from Water Sources and Diarrhea Cases by Random Amplified Polymorphic DNA in Basrah Governorate

Author(s): Zeena hashim Abd Al Wahid, Munaff Jawdat Abd Al-Abbas

One hundred samples of water and thirty of diarrheal cases were collected . Coliform bacteria was detected in most water samples by MPN to comparison with clinical coliform , tap water showed the higher rates of coliform than other sources . 23 different bacterial species from water and 3 from diarrhea were identified by 16SrDNA . E.coli 26 ( 35% ) as the prodomient species from water and 27 (79%) from diarrhea followed by K. pneumonia 11 (14.1%) and 4 (11.8%) respectively. Antibiotic susceptibility showed all E.coli isolates (100%) were MDR and the Ciprofloxacin was the more active antibiotic against isolates of water ( 59% ) and clinics ( 38% ) . In addition , the antibiotic susceptibility profiles showed the identical relatedness of 22 (53.65%) E.coli strains from water and diarrhea . Moreover , RAPD analysis showed 14 (34.14%) of E.coli strains from water and diarrhea were identical indicating to the fecal contamination of water.

E.coli, RAPD, Fecal Contamination, Diarrhea, 16S rDNA, Strains

1. Ahmed, T. A. S. N. I. A., Acharjee, M., Rahman, M. S., Meghla, M., Jamal, J., Munshi, S. K., & Noor, R. A. S. H. E. D. (2013). Microbiological study of drinking water: qualitative and quantitative approach. Asian Jr. of Microbiol. Biotech. Env. Sc, 15(4), 23-458.
2. Ahmed, W., R. Neller and M. Katouli, (2005). Host species-specific metabolic fingerprint Database for Enterococci and Escherichia coli and its application to identify sources of fecal contamination in surface waters. Applied Environ. Microbiol., 71: 4461-4468
3. Bauer, A. W., Kirby, W. M. M., Sherris, J. C., & Turck, M. (1966). Antibiotic susceptibility testing by a standardized single disk method. American journal of clinical pathology, 45(4_ts), 493-496.
4. Bosshard, P. P., Zbinden, R., Abels, S., Böddinghaus, B., Altwegg, M., & Böttger, E. C. (2006). 16S rRNA gene sequencing versus the API 20 NE system and the VITEK 2 ID-GNB card for identification of non fermenting Gram-negative bacteria in the clinical laboratory. Journal of clinical microbiology, 44(4), 1359-1366.
5. Cabral, J. P. (2010). Water microbiology. Bacterial pathogens and water. International journal of environmental research and public health, 7(10), 3657-3703.
6. Campana, R., Scesa, C., Patrone, V., Vittoria, E. and Baffone,W. (2006) Microbiological study of cosmetic products during their use by consumers: health risk efficacy of preservative systems. Lett Appl Microbiol 43, 301–306
7. Cappuccino, J. G., & Sherman, N. (1996). Microbiology: a laboratory manual. Publishing Co., Inc., Menlo Park, California
8. Chao, K. K., Chao, C. C., & Chao, W. L. (2003). Suitability of the traditional microbial indicators and their enumerating methods in the assessment of fecal pollution of subtropical freshwater environments. Journal of Microbiology, Immunology and Infection, 36(4), 288-293.
9. Farris, J. S. (1982). "Out groups and Parsimony". Systematic Biology. 31 (3): 328–334. doi:10.1093/sysbio/31.3.328. ISSN 1063-5157.
10. Fawell , J. and Nieuwenhuijsen , M.J. (2003).Contaminants in drinking water. Br. Med. Bull.68:199-208.
11. Garba, I., M.B. Tijjani, M.S. Aliyu, S.E. Yakubu, A. Wada-Kura and O.S. Olonitola,( 2009).Prevalence of Escherichia coli some public water sources in Gusau Municipal, North Western Nigeria. Bayero J. Pure Applied Sci., 2: 134-137.
12. Garcia-Vallve S. and Puigbo P. (1999-2009). "Dendo UPGMA: A dendrogram construction utility", Universal Rovira i Virgili (URV).Tarragona. Spain.
13. Gassama-Sow, A., Diallo, M. H., Wane, A. A., Seck, A., Samb-Ba, B., Sow, P. S., & Aïdara-Kane, A. (2010). Genetic determinants of antibiotic resistance in diarrheagenic Klebsiella pneumoniae subspecies ozaenae: an emerging enteropathogen in Senegal. Clinical infectious diseases, 50(3), 453-454.
14. Gavriel, A. A., Landre, J. P. B., & Lamb, A. J. (1998). Incidence of mesophilic Aeromonas within a public drinking water supply in north‐east Scotland. Journal of Applied Microbiology, 84(3), 383-392.
15. Girdoniya, M. V. (2011) MPN Test to Detect the Presence of Coliforms in Pond Water and Potable Water.
16. Heiman KE & Bowen A (2013). Shigellosis. In: Travelers’ Health (Ed.). Infectious Diseases Related to Travel. Centers for Disease Control and Prevention, 1600 Clifton Rd. Atlanta, GA, USA. 30: 333.
17. Hidron, A. I., Edwards, J. R., Patel, J., Horan, T. C., Sievert, D. M., Pollock, D. A., & Fridkin, S. K. (2008). Antimicrobial-resistant pathogens associated with healthcare-associated infections: annual summary of data reported to the National Healthcare Safety Network at the Centers for Disease Control and Prevention, 2006–2007. Infection Control & Hospital Epidemiology, 29(11), 996-1011
18. Janda, J. M., & Abbott, S. L. (2006). The Genera Klebsiella and Raoultella.The Enterobacteria (2nd ed., pp. 115-129). Washington, USA: ASM Press.
19. Kaper, J.B., Nataro, J.P., Mobley, H.L.(2004) Pathogenic Escherichia coli. Nat Rev Microbial , 2:123‑40 .
20. Kumar, D., Malik, S., Madan, M., Pandey, A., & Asthana, A. K. (2013). Bacteriological analysis of drinking water by MPN method in a tertiary care hospital and adjoining area Western UP, India. J Environ Sci, 4, 17-22.
21. Lee, M. F., Peng, C. F., Lin, Y. H., Lin, S. R., & Chen, Y. H. (2008). Molecular diversity of class 1 integrons in human isolates of Aeromonas spp. from southern Taiwan. Jpn J Infect Dis, 61(5), 343-349.
22. Livermore, D. M., Winstanley, T. G., & Shannon, K. P. (2001). Interpretative reading: recognizing the unusual and inferring resistance mechanisms from resistance phenotypes. Journal of Antimicrobial Chemotherapy, 48(suppl_1), 87-102.
23. Lu, M. C., Chen, Y. T., Chiang, M. K., Wang, Y. C., Hsiao, P. Y., Huang, Y. J., ... & Lai, Y. C. (2017). Colibactin contributes to the hypervirulence of pks+ K1 CC23 Klebsiella pneumoniae in mouse meningitis infections. Frontiers in cellular and infection microbiology, 7, 103.
24. Magiorakos, A. P., Srinivasan, A., Carey, R. B., Carmeli, Y., Falagas, M. E., Giske, C. G., ... & Paterson, D. L. (2012). Multidrug‐resistant, extensively drug‐resistant and pandrug‐resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clinical microbiology and infection, 18(3), 268-281.
25. Maiti, B., Shekar, M., Khushiramani, R., Karunasagar, I., & Karunasagar, I. (2009). Evaluation of RAPD-PCR and protein profile analysis to differentiate Vibrio harveyi strains prevalent along the southwest coast of India. Journal of genetics, 88(3), 273-279.
26. Miyoshi, T., Iwatsuki, T., & Naganuma, T. (2005). Phylogenetic characterization of 16S rRNA gene clones from deep-groundwater microorganisms that pass through 0.2-micrometer-pore-size filters. Applied and environmental microbiology, 71(2), 1084-1088.
27. Muringani, B. N., Obi, C. L., Apalata, T., & Vasaikar, S. D.( 2016 ) . Bacterial Etiology Of Diarrheal Syndrome And Susceptibility Profile Of Enteropathogens Among Patient Attending Mthata General Hospital. 3 (3 ) .
28. Nielsen, K. L., Godfrey, P. A., Stegger, M., Andersen, P. S., Feldgarden, M., & Frimodt-Møller, N. (2014). Selection of unique Escherichia coli clones by random amplified polymorphic DNA (RAPD): Evaluation by whole genome sequencing. Journal of microbiological methods, 103, 101-103.
29. Obi , C.L., Ramalivhana , J., Samie ,A., Igumbor , E.O. (2007) Prevalence pathogenesis, and antibiotic susceptibility profiles of Aeromonas isolatesfrom stool samples of patients in the Venda region of South Africa. J Health Popul Nutr;25:428-35.
30. Pitout, J. D. D., Revathi, G., L Chow, B., Kabera, B., Kariuki, S., Nordmann, P., & Poirel, L. (2008). Metallo‐β‐lactamase‐producing Pseudomonas aeruginosa isolated from a large tertiary centre in Kenya. Clinical Microbiology and Infection, 14(8), 755-759.
31. Probert, W. S., McQuaid, C., & Schrader, K. (2014). Isolation and identification of an Enterobacter cloacae strain producing a novel subtype of Shiga toxin type 1. Journal of clinical microbiology, JCM-00338.
32. Probert, W. S., McQuaid, C., & Schrader, K. (2014). Isolation and identification of an Enterobacter cloacae strain producing a novel subtype of Shiga toxin type 1. Journal of clinical microbiology, JCM-00338.
33. Ramalivhana, J. N., Obi, C. L., Samie, A., Labuschagne, C., & Weldhagen, G. F. (2010). Random amplified polymorphic DNA typing of clinical and environmental Aeromonas hydrophila strains from Limpopo province, South Africa. Journal of health, population, and nutrition, 28(1), 1.
34. Reller, L. B., Weinstein, M., Jorgensen, J. H., & Ferraro, M. J. (2009). Antimicrobial susceptibility testing: a review of general principles and contemporary practices. Clinical infectious diseases, 49(11), 1749-1755.
35. Schmidt, M. A. (2010). LEE ways: tales of EPEC, ATEC and EHEC. Cellular microbiology, 12(11), 1544-1552
36. Slutsker, L., Ries, A. A., Greene, K. D., Wells, J. G., Hutwagner, L., & Griffin, P. M. (1997). Escherichia coli O157: H7 diarrhea in the United States: clinical and epidemiologic features. Annals of internal medicine, 126(7), 505-513.
37. Tharannum, S., Sunitha, S., Nithya, J., Chandini, M., Vanitha, J., Manjula, T. S., & Shyam Sundar, C. (2009). Molecular confirmation of the presence of coliforms in drinking water using polymerase chain reaction. Kathmandu Univ J Sci Eng Technol, 5(2), 130-136.
38. United Nation Children’s Fund (UNICEF) (2014) World Water Day 2025: 4,000 children die each day from a lack of safe water., http://www.unicef.org/wash/ index_25637.htm
39. WHO (2010) Millennium Development Goals: progress towards the health-related Millennium Development Goals.,http://www.who.int/mediacentre/factsheets/fs290/en/index.html.