Open Access
Research
(Published
online: 11-07-2016)
5.
Genetic characterization of fluoroquinolone-resistant
Escherichia coli
associated with bovine mastitis in India -
Sangeetha Balakrishnan, Prabhakar Xavier Antony, Hirak Kumar
Mukhopadhyay, Raghavan Madhusoodanan Pillai, Jacob Thanislass,
Vijayalakshmi Padmanaban and Mouttou Vivek Srinivas
Veterinary World, 9(7): 705-709
doi:
10.14202/vetworld.2016.705-709
Sangeetha Balakrishnan:
Department of Veterinary Microbiology, Rajiv Gandhi Institute of
Veterinary Education and Research, Puducherry - 605 009, India;
sangeethabvs@gmail.com
Prabhakar Xavier Antony:
Department of Veterinary Microbiology, Rajiv Gandhi Institute of
Veterinary Education and Research, Puducherry - 605 009, India;
pxantony@gmail.com
Hirak Kumar Mukhopadhyay:
Department of Veterinary Microbiology, Rajiv Gandhi Institute of
Veterinary Education and Research, Puducherry - 605 009, India;
mhirak@rediffmail.com
Raghavan Madhusoodanan Pillai:
Department of Veterinary Microbiology, Rajiv Gandhi Institute of
Veterinary Education and Research, Puducherry - 605 009, India;
drrmpillai@gmail.com
Jacob Thanislass:
Department of Veterinary Biochemistry, Rajiv Gandhi Institute of
Veterinary Education and Research, Puducherry - 605 009, India;
jthanislass@gmail.com
Vijayalakshmi Padmanaban:
Department of Veterinary Medicine, Rajiv Gandhi Institute of
Veterinary Education and Research, Puducherry - 605 009, India;
vijinethira@gmail.com
Mouttou Vivek Srinivas:
Department of Veterinary Microbiology, Rajiv Gandhi Institute of
Veterinary Education and Research, Puducherry - 605 009, India;
vivekvet24@gmail.com
Received: 11-01-2016, Accepted: 03-06-2016, Published online:
11-07-2016
Corresponding author:
Mouttou Vivek Srinivas, e-mail: vivekvet24@gmail.com
Citation:
Balakrishnan S, Antony PX, Mukhopadhyay HK, Pillai RM,
Thanislass J, Padmanaban V, Srinivas MV (2016) Genetic
characterization of fluoroquinolone-resistant Escherichia
coli associated with bovine mastitis in India, Veterinary
World, 9(7): 705-709.
Abstract
Aim:
The present study was undertaken to characterize the mutation in
gyrA
(DNA gyrase) and
parC
(topoisomerase IV) genes responsible for fluoroquinolone
resistance in
Escherichia coli
isolates associated with the bovine mastitis.
Materials and Methods:
A total of 92 milk samples from bovine mastitis cases were
sampled in and around Puducherry (Southern India). Among these
samples, 30 isolates were bacteriologically characterized as
E. coli.
Minimum inhibitory concentrations (MIC) of fluoroquinolones of
these 30
E. coli
isolates were evaluated by resazurin microtiter assay. Then, the
quinolone resistance determining region (QRDR) (gyrA
and
parC
genes) of these
E. coli
isolates was genetically analyzed for determining the
chromosomal mutation causing fluoroquinolone resistance.
Results:
E. coli
isolates showed a resistance rate of 63.33%, 23.33% and 30.03%
to nalidixic acid, ciprofloxacin and enrofloxacin, respectively.
Mutations were found at 83rd and 87th amino acid position of
gyrA
gene, and at 80th and 108th amino acid position of
parC
gene in our study isolates. Among these five isolates, one had a
single mutation at 83 amino acid position of
gyrA
with reduced susceptibility (0.5 μg/ml) to ciprofloxacin. Then,
in remaining four isolates, three isolates showed triple
mutation (at
gyrA:
S83àL
and D87àN;
at
parC:
S80àI)
and the fifth isolate showed an additional mutation at codon 108
of
parC
(A108àT)
with the increased ciprofloxacin MIC of 16-128 μg/ml. The most
common mutation noticed were at S83àL
and D87àN
of
gyrA
and S80àI
of
ParC.
Conclusion:
The study confirms the presence of mutation/s responsible for
fluoroquinolone resistance in QRDR of
gyrA
and
parC
genes of
E. coli
isolates of animal origin, and there is increased rate of
fluoroquinolone resistance with high-level of MIC. The mutations
observed in this study were similar to that of human isolates.
Keywords:
Escherichia coli,
fluoroquinolones,
gyrA,
parC,
quinolone resistance determining region.
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