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 AiM  Vol.6 No.7 , June 2016
Detection and Transfer of Extended Spectrum Beta Lactamase Enzymes from Untreated Hospital Waste Water
Abstract: Untreated Hospital wastewater piped into septic tanks contributes to the spread of antibiotic resistance in developing countries. This study was conducted to determine the resistant profile, and Extended Spectrum Beta-Lactamases (ESBLs) production in isolates from hospital waste water, of 2 hospitals in Delta State, Nigeria. A total of 147 organisms were isolated from 32 waste water samples. One hundred and twenty three isolates were Gram negative and 24 were Gram positive. Escherichia coli was the most prevalent in the two locations. Antimicrobial susceptibility by standard disk diffusion method was performed. All isolates were resistant to 4 or more antimicrobial agents. Out of the 123 Gram negative Bacteria, 33 were pan drug resistant and were selected for plasmid curing, DNA extraction and phenotypic detection of ESBL. Transfer of resistant by broth mating technique was performed. Plasmid curing and extraction result indicated that isolates carried resistance on the plasmid and harboured similar multiple high molecular weight plasmids of 23.13 kb and 9.4 kb. ESBL production was detected in 15.15%. Transfer of resistant genes between ESBL producing and non-ESBL producing isolates was observed. Incidence of ESBL in untreated hospital waste water has public health implications. Therefore establishment of treatment plants in our hospital is paramount in achieving sustainable health.
Cite this paper: Egbule, O. (2016) Detection and Transfer of Extended Spectrum Beta Lactamase Enzymes from Untreated Hospital Waste Water. Advances in Microbiology, 6, 512-520. doi: 10.4236/aim.2016.67051.
References

[1]   Chitnis, V., Chitnis, D.O., Patil, S. and Ravi Kant, S. (2000) Hospital Effluent a Source of Multiple Drug Resistant Bacteria. Current Science, 79, 989-991.

[2]   Chitnisa, V., Chitinisa, S., Vaidyaa, K., Ravikants, S., Patilb, S. and Chintinisa, D.S. (2004) Bacterial Population Changes in Hospital Effluent Treatment Plant in Central India. Water Research, 38, 441-447.
http://dx.doi.org/10.1016/j.watres.2003.09.038

[3]   Schwartz, T., Kohnen, W., Jansen, B. and Obst, U. (2003) Detection of Antibiotic-Resistant Bacteria and Their Resistance Genes in Waste Water, Surface Water and Drinking Waterbiofilms. FEMS Microbiology Ecology, 43, 325-335.

[4]   Truong, P. (2000) Vetiver System for Water Quality Improvement. 3rd International Conference of Vetiver and Water, 61-72.

[5]   Xia, H.P., Ke, H.H., Deng, Z.P., Tan, P. and Liu, S.Z. (2004) Ecological Effectiveness of Vetiver Constructed Wetlands in Treating Oil Refined Wastewater. 3rd International Conference of Vetiver and Water.

[6]   Kummerer, K. (2004) Resistance in the Environment. Journal of Antimicrobial Chemotherapy, 54, 311-320.
http://dx.doi.org/10.1093/jac/dkh325

[7]   Harwood, V.J., Bronwell, M., Perusek, W. and Whitlock, J.E. (2001) Vancomycin-Resistant Enterococcus spp. Isolated from Wastewater and Chicken Feces in the United State. Applied and Environmental Microbiology, 67, 4930-4933.
http://dx.doi.org/10.1128/AEM.67.10.4930-4933.2001

[8]   Iversen, A., Franklin, A. and Kuhn, I. (2002) High Prevalence of Vacomycin-Resistant Enterococci in Swedish Sewage. Journal of Applied and Environmental Microbiology, 6, 2838-2842.
http://dx.doi.org/10.1128/AEM.68.6.2838-2842.2002

[9]   Blanch, A.R., Caplin, J.L., Iversen, A., Kuhn, I., Manero, A., Taylor, H.D. and Vilanpva, X. (2003) Comparison of Enterococcal Populations Related to Urban and Hospital Wastewater in Various Climatic and Geographic European Regions. Journal of Applied Microbiology, 51, 1285-1292.

[10]   Clinical and Laboratory Standards Institute (2009) Percformance Standards for Antimicrobial Susceptibility Testing: Seventeenth Informational Supplement. CLSI Document M100-517. Clinical and Laboratory Standards Institute, Wayne.

[11]   Mirmomeni, M.H., Colagar, A.H. and Ghazaey, S. (2007) Molecular Study of Salmonella enteritidis in Poultry Samples by Per, Plasmid Curing, Antibiotic Resistance and Protein Pattern Analysis. Pakistan Journal of Biological Sciences, 10, 1562-1570.
http://dx.doi.org/10.3923/pjbs.2007.1562.1570

[12]   Birnboim, H.C. and Doly, J. (2014) A Rapid Alkaline Extraction Procedure for Screening Recombinant Plasmid DNA. Nucleic Acids Research, 7, 1513-1523.
http://dx.doi.org/10.1093/nar/7.6.1513

[13]   Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Susceptibility Testing: 20th Informational Supplement. CLSI Document VET01-A4 and VET01S-Ed3, CLSI, Wayne.

[14]   Chen, L., Chen, Z.L., Lin, J.H., Zeng, Z.L., Ma, J.Y. and Jiang, H.X. (2007) Emergence of RmtB Methylase-Producing Escherichia coli and Enterobacter Cloacae Isolates from Pigs in China. Journal of Antimicrobial Chemotherapy, 59, 880-885.

[15]   Iroha Iroha, I.R., Amadi, E.S., Nwazo, A.C. and Ejike-Ugwu, P.C. (2010) Detection of Plasmid Borne ESBLs from Blood and Urine Isolates of Gram Negative Bacteria from University Teaching Hospital in Nigeria. Current Research in Bacteriology, 3, 77-83.
http://dx.doi.org/10.3923/crb.2010.77.83

[16]   Muela, A., Pocino, I. and Arana, J. (1994) Effects of Growth Phase and Parental Cell Survival in River Water on Plasmid Transfer between Escherichia coli Strains. Applied and Environmental Microbiology, 60, 4273-4278.

[17]   Saini, S., Das, B.K., Kapil, A., Nagarajan, S.S. and Sarma, R.K. (2004) The Study of Bacterial Flora of Different Types in Hospital Waste: Evolution of Waste Treatment at AIIMS Hospital, New Delhi. The Southeast Asian Journal of Tropical Medicine and Public Health, 35, 986-989.

[18]   Abdulaziz, Y.A. (2011) Review on Hospital Wastes and Its Possible Treatments. Egyptian Academic Journal of Biological Sciences, 3, 55-62.

[19]   Horii, T., Arakawa, Y., Ohta, M., Ichiyama, S., Wacharotayankun, R. and Kato, N. (1993) Plasmid Mediated AmpC-Type β-Lactamase Isolated from Klebsiella pneumoniae Confers Resistance to Broad Spectrum β-Lactams Including Moxalactam. Antimicrobial Agents and Chemotherapy, 37, 984-990.
http://dx.doi.org/10.1128/AAC.37.5.984

[20]   Molstad, S., Lundborg, C.S. and Karlsson, A.K. (2002) Antibiotic Prescription Rates Vary Markedly between 13 European Countries. Scandinavian Journal of Infectious Diseases, 34, 366-371.
http://dx.doi.org/10.1080/00365540110080034

[21]   Davoud, K. and Shahla, M. (2011) Emergence of Multiple β-Lactamases Produced by Escherichia coli Clinical Isolates from Hospitalized Patient in Kerman, Iran. Jundishapur Journal of Microbiology, 3, 137-145.

[22]   Jean, S.S., Teng, L.J., Hsueh, P.R., Ho, S.W. and Luh, K.T. (2002) Antimicrobial Susceptibility among Clinical Isolates of Extended-Spectrum Cephalosporin-Resistant Gram-Positive Bacteria in a Taiwanese University Hospital. Journal of Antimicrobial Chemotherapy, 49, 69-76.
http://dx.doi.org/10.1093/jac/49.1.69

[23]   Rawat, D. and Nair, D. (2010) Extended-Spectrum β-Lactamases in Gram Negative Bacteria. Journal of Global Infectious Diseases, 2, 263-274.

[24]   Singhal, S., Mathur, T., Khan, S., Upadhyay, D.J., Chugh, S., Gaind, R. and Rattan, A. (2005) Evaluation of Methods for AmpC Beta-Lactamase in Gram Negative Clinical Isolates from Tertiary Care Hospitals. Indian Journal of Medical Microbiology, 23, 120-124.
http://dx.doi.org/10.4103/0255-0857.16053

[25]   Reich, F., Atanassova, V., & Gunter, K. (2013) Extended-Spectrum β-Lactamase and AmpC-Producing Enterobacteria in Healthy Broiler Chickens, Germany. Emerging Infectious Disease, 19, 1253-1258.
http://dx.doi.org/10.3201/eid1908.120879

[26]   Yusuf, I., Yusha’u, M., Getso, M., Yahaya, H., Bala, J., Aliyu, I. and Haruna, M. (2012) Detection of Metallo-Beta-Lactamases among Gram Negative Bacterial Isolates from Murtala Muhammed Specialist Hospital, Kano and Almadina Hospital Kaduna, Nigeria. Bayero Journal of Pure and Applied Sciences, 5, 84-88.

[27]   Bunny, K.L., Hall, R.M. and Stokes, H.W. (1995) New Mobile Gene Cassettes Containing an Aminoglycoside Resistance Gene, aacA7, and a chloramphenicol Resistance Gene, catB3, in an Integron in pBWH301. Antimicrobial Agents and Chemotherapy, 39, 686-693.
http://dx.doi.org/10.1128/AAC.39.3.686

[28]   Murray, B.E. (1997) Antibiotic Resistance. Journal of Advances in Internal Medicine, 42, 339-367.

 
 
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