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. 2017 May;10(5):517-520.
doi: 10.14202/vetworld.2017.517-520. Epub 2017 May 12.

Detection of emerging antibiotic resistance in bacteria isolated from subclinical mastitis in cattle in West Bengal

Arnab Das  1 , Chanchal Guha  2 , Ujjwal Biswas  2 , Partha Sarathi Jana  2 , Amaresh Chatterjee  2 , Indranil Samanta  3
Affiliations

Detection of emerging antibiotic resistance in bacteria isolated from subclinical mastitis in cattle in West Bengal

Arnab Das et al. Vet World. 2017 May.

Abstract

Aim: The aim of this work was to detect antibiotic resistance in Gram-negative bacteria isolated from subclinical mastitis in cattle in West Bengal.

Materials and methods: The milk samples were collected from the cattle suffering with subclinical mastitis in West Bengal. The milk samples were inoculated into the nutrient broth and incubated at 37°C. On the next day, the growth was transferred into nutrient agar and MacConkey agar. All the pure cultures obtained from nutrient agar slant were subjected to Gram-staining and standard biochemical tests. All the bacterial isolates were tested in vitro for their sensitivity to different antibiotics commonly used in veterinary practices. All Gram-negative isolates including positive control were subjected to polymerase chain reaction (PCR) for detection of blaCTX-M, blaTEM, blaSHV, blaVIM, tetA, tetB, tetC, and tetM genes considered for extended-spectrum β-lactamase (ESBL), metallo-β-lactamase, and tetracycline resistance.

Results: In total, 50 Gram-negative organisms (Escherichia coli, Proteus, Pseudomonas, Klebsiella, and Enterobacter) were isolated from milk samples of subclinical mastitis infected cattle. Among these Gram-negative isolates, 48% (24/50) were found either ESBL producing or tetracycline resistant. Out of total 50 Gram-negative isolates, blaCTX-M was detected in 18 (36%) isolates, and 6 (12%) harbored blaTEM genes in PCR. None of the isolates carried blaSHV genes. Further, in this study, 5 (10%) isolates harbored tet(A) gene, and 8 (16%) isolates carried tet(B) gene. No tet(C) gene was detected from the isolates.

Conclusion: This study showed emerging trend of antibiotic-resistant Gram-negative bacteria associated with subclinical mastitis in cattle in West Bengal, India.

Keywords: Escherichia coli; India; antibiotic resistance; cattle; sub-clinical mastitis.

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Figures

Figure-1
Figure-1
Gel electrophoresis image of polymerase chain reaction amplified products of blaCTX-M gene of Gram-negative isolates; Lane 1 - 1500 bp DNA ladder; Lane 2-14, 16-19 - Isolated Escherichia coli samples; Lane 15 - Positive control for blaCTX-M gene; Lane 20 - Negative control.
Figure-2
Figure-2
Gel electrophoresis image of polymerase chain reaction amplified products of blaTEM gene of Gram-negative isolates; Lane 1 - 1500 bp DNA ladder; Lane 2-8, 10-16 - Isolated Escherichia coli samples; Lane 15 - Positive control for blaTEM gene; Lane 20 - Negative control.
Figure-3
Figure-3
Gel electrophoresis image of polymerase chain reaction amplified products of tetA gene of Gram-negative isolates; Lane 1 - 1500 bp DNA ladder; Lane 2-16, 18-19 - Isolated Escherichia coli samples; Lane 17 - Positive control for tetA gene; Lane 20 - Negative control.
Figure-4
Figure-4
Gel electrophoresis image of polymerase chain reaction amplified products of tetB gene of Gram-negative isolates. Lane 1 - 1500 bp DNA ladder; Lane 2-14, 16-19 - Isolated Escherichia coli samples; Lane 15 - Positive control for tetB gene; Lane 20 - Negative control.
See this image and copyright information in PMC

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