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Choi, Lee, Kim, Kim, Park, Bae, Cho, Kim, Kim, and Kang: Antibiotic Resistance and Genotypic Characterization of Escherichia coli Isolated from Pig Farm Environment
Original Article

J Bacteriol Virol 2006;36(3):159-165.

Published online: 18 February 2006

DOI: https://doi.org/10.4167/jbv.2006.36.3.159

Antibiotic Resistance and Genotypic Characterization of Escherichia coli Isolated from Pig Farm Environment

Seung-Hwa Choi1, Young-Ju Lee2, Bong-Hwan Kim2, Ki-Seuk Kim2, Cheong-Kyu Park2, Dong Hwa Bae2, Jae-Keun Cho3, Jong-Wan Kim4, Byoung-Han Kim4, Min-Su Kang4

1Busan Metropolitan City Research Institute of Health & Environment, Busan, 613-806, Korea

2College of Veterinary Medicine, Kyungpook National University, Daegu, 702-701, Korea

3Daegu Metropolitan City Research Institute of Health & Environment, Daegu, 706-732, Korea

4National Veterinary Research and Quarantine Service, Anyang, 430-824, Korea

Received 20 June 2006       Accepted 9 August 2006

Copyright © 2006 Journal of Bacteriology and Virology

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The use of antibiotics, including therapeutically in human and veterinary medicine, or as prophylaxis of growth promotion in animal husbandry, ultimately exerts selective pressure favorable for the propagation of antibiotic resistant bacteria. In this study we have determined the resistance for antibiotics of E. coli from pig farm environment, and investigate genetic relatedness by random amplification of polymorphic DNA (RAPD). Six farms were randomly selected in Gyeongsanman-do and Busan provinces for collecting samples from feces, manure and underground water. A total of 88 isolates from feces, 74 isolates from manure and 1 isolate from underground water were analyzed by antibiotic resistance and RAPD. Antibiotic resistance testing was performed by disk diffusion method using 16 antibiotics. The highest percentage of antibiotic resistance of isolates from feces and manure was found to the following antibiotics; tetracycline (100% and 100%), sulfamethoxazole/trimethoprim (60.2% and 62.2%), streptomycin (50.0% and 68.9%), chloramphenicol (56.8% and 56.8%), ampicillin (50.0% and 81.1%) and cephalothin (50.0% and 51.4%). Of isolates from feces and manure, 22.7% and 20.3% showed multiple resistance to 4 and 5 antibiotics, respectively. The isolates from GE pig farm showed six RAPD patterns. A single pattern, RAPD-C, was predominat in feces isolates (50.0%) and manual isolates (46.7%), and the rest of feces isolates showed RADP-A, B and E pattern and manure isolates showed D and E pattern. One isolate from underground water showed F pattern. The appearance of multiresistant in E. coli isolates from pig farms environment is a problem of major concern of public health and RAPD may offer an useful tool of discrimination for the epidemiological investigation.

Keywords: Escherichia coli, Antibiotic resistance, Randomly amplification of polymorphic DNA

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Figure 1.
Distribution of multiresistant E. coli isolates.
jbv-36-159f1.tif
Figure 2.
Representative RAPD fingerprinting profiles of E. coli isolates from E farm. Lane MW contains the 200 bp ladder molecular size marker.
jbv-36-159f2.tif
Table 1.
Isolation frequency of E. coli from pig farm environment
Farm Province No. of pigs housed Feces Manure Underground water
No. of sample No. of isolates No. of sample No. of isolates No. of sample No. of isolates
GA Busan 500 10 14 10 12 2 0
GB Busan 1,000 10 13 10 10 2 0
GC Busan 500 10 11 10 1 1 0
GD Jinju 2,000 10 18 10 19 1 0
GE Sacheon 1,000 10 20 10 15 1 1
GF Kimhae 800 10 12 10 17 1 0
Total – – 60 88 60 74 8 1
Table 2.
Antimicrobial resistance of E. coli isolates
Sample No. of isolates tested Type∗∗ No. (%) of resistant isolates∗
Am Amc Cf Cz Fox Ctx Fep Ipm S Gm An Cip Nor Sxt Cp Te
Feces 88 R 44 (50.0) 2 (2.3) 44 (50.0) 3 (3.4) 1 (1.1) 1 (1.1) 1 (1.1) 0 (0.0) 44 (50.0) 11 (12.5) 1 (1.1) 9 (10.2) 7 (8.0) 53 (60.2) 50 (56.8) 88 (100)
I 10 (11.4) 12 (13.6) 37 (42.0) 31 (35.2) 21 (23.9) 44 (50.0) 0 (0.0) 0 (0.0) 33 (37.5) 12 (13.6) 11 (12.5) 14 (15.9) 1 (1.1) 4 (4.5) 4 (4.5) 0 (0.0)
Manure 74 R 60 (81.1) 7 (9.5) 38 (51.4) 12 (16.2) 11 (14.9) 2 (2.7) 6 (8.1) 0 (0.0) 51 (68.9) 16 (21.6) 2 (2.7) 17 (23.0) 13 (17.6) 46 (62.2) 42 (56.8) 74 (100)
I 4 (5.4) 34 (45.9) 32 (43.2) 36 (48.6) 19 (25.7) 41 (55.4) 0 (0.0) 0 (0.0) 17 (23.0) 15 (20.3) 11 (14.9) 13 (17.6) 9 (12.2) 4 (5.4) 6 (8.1) 0 (0.0)
Underground water 1 R 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 0.0 1 (100) 0 0.0 0 (0.0) 0 0.0 0 0.0 0 0.0 1 (100.) 1 (100.)
I 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 1 (100.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0)

∗ AM, ampicillin; Amc, amoxicillin/clavulanic acid; Cf, cephalothin; Cz, cefozolin; Fox, cefoxitin; Ctx, cefotaxime; Fep, cefepime; Ipm, imipenem; S, streptomycin; Gm, gentamicin; An, amikacin; Cip, ciprofloxacin; Nor, norfloxacin; Sxt, trimethoprim/sulfamethoxazole; Em, erythromycin; Cp, chloramphenicol; TE, tetracycline.

∗∗ R, Resistance; I, Intermediate

Table 3.
Distribution of RAPD fingerprinting profiles of E. coli isolates from E farm
Fingerprinting type No. (%) of isolates
Feces Manure Underground water
A 2 (10.0) 0 (0) 0 (0)
B 6 (30.0) 0 (0) 0 (0)
C 10 (50.0) 7 (46.7) 0 (0)
D 0 (0) 5 (33.3) 0 (0)
E 2 (10.0) 3 (20.0) 0 (0)
F 0 (0) 0 (0) 1 (100)
Total 20 (100) 15 (100) 1 (100)
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