2017-IJDR C.freundii PDF
2017-IJDR C.freundii PDF
2017-IJDR C.freundii PDF
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1Student, Master Program of Medicine and Veterinary Public Health, Faculty of Veterinary Medicine,
Universitas Airlangga
2Department of Veterinary Public Health, Faculty of Veterinary Medicine, Universitas Airlangga
3Department of Veterinary Microbiology, Faculty of Veterinary Medicine, Universitas Airlangga
Article History: This study aims to isolate, to identify, and to seek out fragments of encoding gene Extended
Received 22nd August 2017 Spectrum β-Lactamase on Escherichia coli isolated from swab surface of broiler chicken meat in
Received in revised form a number of traditional markets in Surabaya. The result shows that 31 out of 50 samples
07th September, 2017 positively contain Escherichia coli, shown through EMBA isolation media and identified using
Accepted 16th October, 2017 indole test. Sensitivity test shows that 100% of the isolates are resistant to Ampicilin, 48.4% are
Published online 30th November, 2017 resistant to Cephazoline, 13% are resistant to Ceftazidime, 9.6% are resistant to Cefotaxime, 6.4%
are resistant to Ceftriaxone and 87.2% are resistant to Tetracycline. 8 out of 8 (100%) samples of
Key Words: E. coli resistant show the presence of band towards blaTEM gene of 861basepair (bp).
Extended Spectrum β-Lactamase,
Escherichia coli,
broiler chicken meat,
blaTEM gene,
Antimicrobial Resistance.
Copyright ©2017, Rinda Dewi Safitri et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original work is properly cited.
Citation: Rinda Dewi Safitri, Risi Cicilia, Intan Galuh Bintari, Intan Permatasari Hermawan, Mustofa Helmi Effendi Rahaju Ernawati and Jola
Rahmahani, 2017. “Detection of encoding gene extended spectrum beta lactamase on escherichia coli isolated from broiler chicken meat in traditional
market surabaya”, International Journal of Development Research, 7, (11), 17354-17357.
Testing of Escherichia coli resistant to disc antibiotics Figure 1. The illustration of sensitivity test on Escherichia coli
(OXOID, Basingstoke, United Kingdom), Ampicillin 10 µg isolates towards antibiotics.
(CT0003), Cefotaxime 30 µg (CT0166), Ceftazidime 30 µg
(CT0412), Ceftriaxone 30 µg (CT0417), Cefazolin 30 µg Samples which are resistant to antibiotics in the sensitivity test
(CT0011) and Tetracyclin 30 µg (CT0054), were carried out were then undergone Polymerase Chain Reaction (PCR) in
by disc diffusion method on the medium of Mueler Hinton order to find the fragment of the encoding gene of Extended
Agar (MHA) (E. Merck, Darmstadt, Germany) . Interpretation Spectrum Beta-lactamase (ESBL). 8 out of 8 E. coli resistant
of the result was known through measurement of inhibitory samples show the presence of DNA band from blaTEM gene of
zone diameter formed in accordance with the recommendation 861bp. The electrophoresis result of PCR products can be seen
of the Clinical and Laboratory Standards Institute (CLSI, in Figure 2.
2016).
IDENTIFICATION OF ESBL GENE WITH
POLYMERASE CHAIN REACTION
The initial procedure in conducting PCR technique in this
study was the DNA extraction. DNA extraction of Escherichia
coli used boiling method. PCR amplification of the blaTEM
gene with 20 μl PCR reaction consisted of: 12.5 μL PCR
Master Mix, 1 μL primary forward and 1 μL reverse primer, 5
μL DNA template and 0.5 μL nuclease free water. The mixture
of PCR reagent was then added in the thermocycler. Primer
used and the condition of PCR are presented in Table 1.
Tabel 1. Primer and thermal cycling condition that were used for
amplification of blaTEM gene Figure 2. The electrophoresis result of PCR products indicates
blaTEM gene with band of 861bp
Target
Thermal Cycling
gene Primer used
Condition DISCUSSION
blaTEM F : 5’ GTATCCGC 95°C 15 min → 30x
TCATGGAGA
This study has successfully isolated Escherichia coli by 62%
[95°C 1 min, 58°C 1
CAATAACCCTG-3’ min, 72°C 1 min] → (31 out of 50) samples taken from swab surface of broiler
R : 5’-CCAATGCTT 72°C 10 min → 4°C ∞ chicken meat in 5 traditional markets in the area of Surabaya.
AATCAGTGGAGGCACC-3’ This finding is similar to the study by Bhoomika et al. (2016)
which detects E. coli in chicken meat in Chhattisgrash, India of
RESULT 66.32%. The level of E. coli contamination in chicken meat
reported by Odwar et al. (2014) is also considerably high at
The isolation and identification result on 50 swab samples of 78%.
broiler chicken meat obtained from 5 traditional market in
17356 International Journal of Development Research, Vol. 07, Issue, 11, pp.17354-17357, November, 2017
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