Molecular identification of blaCTX-M and blaTEM genes encoding extended-spectrum ß-lactamase (ESBL) producing Escherichia coli isolated from raw cow’s milk in East Java, Indonesia
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Abstract. Ansharieta R, Ramandinianto SC, Effendi MH, Plumeriastuti H. 2021. Molecular identification of blaCTX-M and blaTEM genes encoding extended-spectrum ß-lactamase (ESBL) producing Escherichia coli isolated from raw cow’s milk in East Java, Indonesia. Biodiversitas 22: 1600-1605. The emergence of extended-spectrum ?-lactamase (ESBL) producing bacteria and its increasing level has become public health issue. The presence of these bacteria in food of animal origin is quite alarming. The objective of this study was to detect and characterize Escherichia coli producing ESBL encoding genes, isolated from 200 raw cow milk samples in East Java, Indonesia. The results of this study showed that 70.5% of isolates were confirmed as E. coli, based on the morphological growth of colonies on the EMB Agar and biochemical IMViC tests. In this study, the double-disc synergy test (DDST) method was used to confirm the ESBL, and previously sorted out presumptively by using Aztreonam antibiotic disc. The antibiotics used were amoxicillin-clavulanate, ceftazidime, and cefotaxime for DDST. In addition, ESBL confirmation with Multiplex PCR method for blaCTX-M and blaTEM genes were done. The presence of ESBL-producing by E. coli isolated from raw cow’s milk in East Java were 2.12% (3/141). The PCR results showed that the double blaCTX-M and blaTEM gene harbored by 2 ESBL isolates and one blaTEM gene as many as 1 ESBL isolate. Thus, the findings of our study indicate that milk can be a good reservoir of bacteria carrying blaCTX-M and blaTEM ESBL resistance genes with the potential to affect human health.
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References
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Wibisono, F.J., Sumiarto, B., Untari, T., Effendi, M.H., Permatasari, D.A., Witaningrum, A.M. 2020. Pattern of antibiotic resistance on extended-spectrum beta-lactamases genes producing Escherichia coli on laying hens in Blitar, Indonesia. Biodiversitas, 21 (10): 4631- 4635.
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Ali, T., Rahman, S., Zhang, L., Shahid, M., Zhang, S., Liu, G., Gao, J., and Han, B. 2016. ESBL Producing Escherichia coli from Cows Suffering Mastitis in China Contain Clinical Class 1 Integrons with CTX-M Linked to ISCR1. Front. Microbiol., 7(11): 1-11.
Batabyal, K., Banerjee, A., Pal, S., Dey, S., Joardar, S.N., Samanta, I., Isore, D.P., and Singh, A.D. 2018. Detection, Characterization, and Antibiogram of Extended-Spectrum Beta-Lactamase Escherichia coli Isolated from Bovine Milk Samples in West Bengal, India. Vet. World, 11 (10): 1423-1427.
Bush, K. (2013) Proliferation and Significance of Clinically Relevant ?-lactamases. Ann. N Y Acad. Sci.1277 (1):84–90.
Calistri P, Iannetti S, Danzetta L, Narcisi M, Cito V, Di Sabatino F. (2013). The components of ‘One World-One Health’ approach. Transbound Emerg Dis. 60: 4-13.
Clinical and Laboratory Standards Institute. [CLSI] 2018. M100 Performance Standards for Antimicrobial Susceptibility Testing. 28th ed. Clinical and Laboratory Standards Institute, USA.
Effendi, M.H., Harijani, N., Budiarto, Triningtya, N.P., Tyasningsih, W. and Plumeriastuti, H. 2019. Prevalence of Pathogenic Escherichia Coli Isolated from Subclinical Mastitis in East Java Province, Indonesia. Indian Vet. J., 96 (03) : 22 - 25
Effendi, M.H., Harijani, N., Yanestria, S.M., and Hastutiek, P. 2018. Identification of Shiga Toxin-Producing Escherichia coli in Raw Milk Samples from Dairy Cows in Surabaya, Indonesia. Philipp. J. Vet. Med., 55(SI): 109-114.
Geser, N., Stephan, R., and Hächler, H. 2012. Occurrence and characteristics of Extended Spectrum ?-Lactamase (ESBL) Producing Enterobacteriaceae in Food Producing Animals, Minced Meat and Raw Milk. BMC Vet. Res., 8 (21): 1-9.
Hinthong, W., Pumipuntu, N., Santajit, S., Kulpeanprasit, S., Buranasinsup, S., Sookrung, N., Chaicumpa, W., Aiumurai, P., and Indrawattana, N. 2017. Detection and Drug Resistance Profile of Escherichia coli from Subclinical Mastitis Cows and Water Supply in Dairy Farms in Saraburi Province, Thailand. PeerJ., 5: 1-16.
Jena, J., Sahoo, R.K., Debata, N.K., and Subudhi, E. 2017. Prevalence of TEM, SHV, and CTX-M Genes of Extended Spectrum ?-Lactamase-Producing Escherichia coli Strains Isolated from Urinary Tract Infections in Adults. 3 Biotech., 7: 1-7.
Kennedy, H., Wilson, S., Marwick, C., Malcolm W., and Nathwani, D. 2015. Reduction in Broad-Spectrum Gram-negative Agents by Diverse Prescribing of Aztreonam within NHS Tayside. J. Antimicrob. Chemother., 2015: 2421-2422.
Kristianingtyas, L., Effendi, M.H., Tyasningsih, W., and Kurniawan, F. 2020. Genetic Identification of blaCTX-M Gene and blaTEM Gene on Extended Spectrum Beta Lactamase (ESBL) Producing Escherichia coli from Dogs. Indian Vet. J., 97(01): 17-21.
Landers, T.F., Cohen, B., Wittum, T.E., and Larson, E.L. 2012. A Review of Antibiotic Use in Food Animals: Perspective, Policy, and Potential. Public Health Rep., 127(1): 4–22.
Odenthal, S., Akineden, Ö., and Usleber, E. 2016. Extended Spectrum ?-Lactamase Producing Enterobacteriaceae in Bulk Tank Milk from German Dairy Farms. Int. J. Food Microbiol., 238: 72-78.
Pereckaite, L., Tatarunas, V., and Giedraitiene, A. 2018. Current Antimicrobial Susceptibility Testing for Beta-Lactamase-Producing Enterobacteriaceae in Clinical Settings. J. Microbiol. Methods, 152: 154-164.
Pitout, J.D.D. (2012) Extraintestinal Pathogenic Escherichia coli: A Combination of Virulence with Antibiotic Resistance. Front. Microbiol. 3(9): 1-7.
Putra, A.R.S., Effendi, M.H., Koesdarto S., and Tyasningsih, W. 2019. Molecular Identification of Extended Spectrum Beta-Lactamase (ESBL) Producing Escherichia coli Isolated from Dairy Cows in East Java Province, Indonesia. Indian Vet. J., 96(10): 26-30.
Putra, A.R. Effendi, M.H. Koesdarto, S. Suwarno, S. Tyasningsih, W. and Estoepangestie, A.T. 2020. Detection of the extended spectrum ?-lactamase produced by Escherichia coli from dairy cows by using the Vitek-2 method in Tulungagung regency, Indonesia. Iraqi Journal of Veterinary Sciences, 34 (1): 203-207.
Rahmahani J, Salamah, Mufasirin, Tyasningsih W Effendi MH. 2020. Antimicrobial Resistance Profile of Escherichia coli From Cloacal Swab of Domestic Chicken In Surabaya Traditional Market. Biochem. Cell. Arch. 20(1): 2993-2997.
Su, Y., Yu, C.Y., Tsai, Y., Wang, S.H., Lee, C., and Chu, C. 2016. Fluoroquinolone Resistant and Extended Spectrum ?-Lactamase (ESBL)-Producing Escherichia coli from Milk of Cow with Clinic Mastitis in Southern Taiwan. J. Microbiol. Immunol., 49: 892-901.
Tanzin, T., Nazir, K.H.M.N.H., Zahan, M.N., Parvej, S.M., Zesmin, K., and Rahman, M.T. 2016. Antibiotic Resistance Profile of Bacteria Isolated from Raw Milk Samples of Cattle and Buffaloes. J. Adv. Vet. Anim. Res., 3(1): 62-67.
Ukah, U.V., Glass, M., Avery, B., Daignault, D., Mulvey, M.R., Reid-Smith, R.J., Parmley, E.J., Portt, A., Boerlin, P., and Manges, A.R. (2017) Risk Factors for Acquisition of Multidrug-Resistant Escherichia coli and Development of Community-Acquired Urinary Tract Infections. Epidemiol. Infect., 146: 46-57.
Villegas, M.V., Correa, A., Perez, F., Zuluaga, T., Radice, M., and Gutkind, G. 2004. CTX-M-12 Beta-Lactamase in A Klebsiella pneumoniae Clinical Isolate in Colombia. Antimicrob. Agents Chemother., 48: 629-631.
Wendt A, Kreienbrock L, Campe A. 2015. .Zoonotic disease surveillance – inventory of systems integrating human and animal disease information. Zoonoses Public Health. 62(1):61-74.
Wibisono, F.J. Sumiarto, B., Untari, T., Effendi, M.H., Permatasari, D.A., Witaningrum. A.M. 2020. The Presence of Extended Spectrum Beta-Lactamase (ESBL) Producing Escherichia coli On Layer Chicken Farms In Blitar Area, Indonesia. Biodiversitas, 21 (6): 2667-2671.
Wibisono, F.J., Sumiarto, B., Untari, T., Effendi, M.H., Permatasari, D.A., Witaningrum, A.M. 2020. CTX Gene of Extended Spectrum Beta-Lactamase (ESBL) Producing Escherichia coli on Broilers in Blitar, Indonesia. Sys Rev Pharm. 11(7): 396-403.
Wibisono, F.J., Sumiarto, B., Untari, T., Effendi, M.H., Permatasari, D.A., Witaningrum, A.M. 2020. Pattern of antibiotic resistance on extended-spectrum beta-lactamases genes producing Escherichia coli on laying hens in Blitar, Indonesia. Biodiversitas, 21 (10): 4631- 4635.
Widodo, A., Effendi, M.H., Khairullah, A.R. 2020. Extended-spectrum beta-lactamase (ESBL)-producing Eschericia coli from livestock. Sys Rev Pharm 11(7): 382-392
Yao, F., Qian, Y., Chen, S., Wang, P., and Huang, Y. (2007) Incidence of Extended-Spectrum Beta-Lactamases and Characterization of Integrons in Extended-Spectrum Beta-Lactamase-producing Klebsiella pneumoniae isolated in Shantou, China. Acta Biochim. Biophys. Sin. (Shanghai) 39 (7): 527-532.
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