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Lee, Choi, Kim, Song, and Yang: Prevalence and Characterization of Actinobacillus pleuropneumoniae Isolated from Korean Pigs
Original Article

J Bacteriol Virol 2015;45(1):19-25.

Published online: 9 February 2015

DOI: https://doi.org/10.4167/jbv.2015.45.1.19

Prevalence and Characterization of Actinobacillus pleuropneumoniae Isolated from Korean Pigs

Ki-Eun Lee1, Hwan-Won Choi2, Ha-Hyun Kim1, Jae-Young Song1, Dong-Kun Yang1, *

1Animal and Plant Quarantine Agency, MAFRA, Anyang, Korea

2ChoongAng Vaccine Laboratories, Daejon, Korea

*Corresponding author: Dong-Kun Yang. Animal and Plant Quarantine Agency, 175 Anyang-ro, Anyang-si, Gyeonggi-do 430-757, Korea. Phone: +82-31-467-1783, Fax: +82-31-467-1797, e-mail: yangdk@korea.kr

** We are grateful to the ChoongAng Vaccine Laboratories for the deposition of A. pleuropneumoniae isolated from pig. This study was supported by a grant (N-AD20-2010-19-01) from the Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs (MAFRA), Republic of Korea.

Received 20 November 20147 January 2015       Accepted 9 January 2015

Copyright © 2015 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

Actinobacillus pleuropneumoniae causes porcine pleuropneumoniae which is one of severe threats to the swine industry. In total, 54 strains of Actinobacillus pleuropneumoniae were isolated from 443 pigs between 2012 and 2013 in Korea. Isolates were classified into serotypes 1, 2, 5, 7, 12, and unclassified by multiplex PCR. Genotypes of isolates were divided into three groups according to the sequence of the omlA gene. The antimicrobial resistance rate of serotype 1 was slightly higher than that of serotype 5. In conclusion, to block and treat porcine pleuropneumonia, it is necessary to conduct ongoing characterization of A. pleuropneumoniae isolated from pigs.

Keywords: Actinobacillus pleuropneumoniae, Serotype, Genotype, Antimicrobial

REFERENCES

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Figure 1.
Alignment among the omlA gene of A. pleuropneumoniae
jbv-45-19f1.tif
Table 1.
PCR condition for serotyping of A. pleuropneumoniae
Name Responsible Primer sequence Size (bp) Ref.
AP1F Forward primer for cps region of serotype 1 5′-AGTGGCTGGATGAGACGAGAC-3′ 1,600 (5)
AP1R Reverse primer for cps region of serotype 1 5′-TAGTTTGTTATGGTATTTCTGTA-3′
AP2F Forward primer for cps region of serotype 2 5′-ACTATGGCAATCAGTCGATTCAT-3′ 500 (10)
AP2R Reverse primer for cps region of serotype 2 5′-CCTAATCGGAAACGCCATTCTG-3′
AP3F Forward primer for cps region of serotype 3 5′-TTTGCGCTGTAGTGCTCCAAT-3′ 900 This study
AP3R Reverse primer for cps region of serotype 3 5′-AACAAATAAAGTTGCTCGAAAGTA-3′
AP5F Forward primer for cps region of serotype 5 5′-TTTATCACTATCACCGTCCACACCT-3′ 1,100 (10)
AP5R Reverse primer for cps region of serotype 5 5′-CATTCGGGTCTTGTGGCTACTAA-3′
AP6F Forward primer for cps region of serotype 6 5′-AACCACTCACTTTCCACATTAG-3′ 720 (10)
AP6R Reverse primer for cps region of serotype 6 5′-AATCGGAAGGTTTTGGTCTCGTG-3′
AP7F Forward primer for cps region of serotype 7 5′-GGTGACTGGCGTACGCCAAA-3′ 400 This study
AP7R Reverse primer for cps region of serotype 7 5′-GGGCTGCAGACTGACGTAA-3′
AP8F Forward primer for cps region of serotype 8 5′-AACGGCTTTTGAACAACTTTATTTATTT-3′ 1,000 (5)
AP8R Reverse primer for cps region of serotype 8 5′-TTCATTCCTAAACTCCGTATTGTCA-3′
AP12F Forward primer for cps region of serotype 12 5′-GGTTCTCCAGATGACTCTGAAA-3′ 600 This study
AP12R Reverse primer for cps region of serotype 12 5′-GCTATTGGATGAAGATGACTCAT-3′
Table 2.
Classification of A. pleuroneumoniae based on omlA gene sequence
Serotype Strain KVCC No. Date Province Genotype (omlA gene)
A. pleuroneumoniae KVCC-BA1400162 2012.07.06. Jeonbuk I
A. pleuroneumoniae KVCC-BA1400164 2012.07.10. Chungbuk I
A. pleuroneumoniae KVCC-BA1400165 2012.07.17. Gyeonggi I
A. pleuroneumoniae KVCC-BA1400171 2012.11.20. Gyeonggi I
A. pleuroneumoniae KVCC-BA1400174 2012.11.27. Chungnam I
A. pleuroneumoniae KVCC-BA1400176 2013.02.15. Chungnam I
A. pleuroneumoniae KVCC-BA1400182 2013.03.15. Chungnam I
1 (n=15) A. pleuroneumoniae KVCC-BA1400193 2013.12.18. Chungnam I
A. pleuroneumoniae KVCC-BA1400196 2012.01.02. Chungnam I
A. pleuroneumoniae KVCC-BA1400213 2012.01.06. Chungbuk I
A. pleuroneumoniae KVCC-BA1400214 2012.01.06. Gyeongnam I
A. pleuroneumoniae KVCC-BA1400221 2012.04.20. Jeonbuk I
A. pleuroneumoniae KVCC-BA1400222 2012.05.02. Chungnam I
A. pleuroneumoniae KVCC-BA1400223 2012.05.02. Chungnam I
A. pleuroneumoniae KVCC-BA1400170 2012.11.20. Chungnam I
A. pleuroneumoniae KVCC-BA1400167 2012.09.03. Gyeongbuk II
A. pleuroneumoniae KVCC-BA1400173 2012.11.21. Chungnam II
2 (n=4) A. pleuroneumoniae KVCC-BA1400181 2013.03.08. Gyeongnam II
A. pleuroneumoniae KVCC-BA1400188 2013.08.28 Chungnam II
A. pleuroneumoniae KVCC-BA1400163 2012.07.10. Gyeongnam III
A. pleuroneumoniae KVCC-BA1400169 2012.09.19. Gyeongbuk III
A. pleuroneumoniae KVCC-BA1400172 2012.11.21. Gyeonggi III
A. pleuroneumoniae KVCC-BA1400175 2012.12.21. Gyeongbuk III
A. pleuroneumoniae KVCC-BA1400178 2013.02.28. Gyeongnam III
A. pleuroneumoniae KVCC-BA1400184 2013.04.08. Chungnam III
A. pleuroneumoniae KVCC-BA1400185 2013.04.24. Chungnam III
A. pleuroneumoniae KVCC-BA1400186 2013.04.24. Chungnam III
A. pleuroneumoniae KVCC-BA1400187 2013.06.07. Gyeongbuk III
A. pleuroneumoniae KVCC-BA1400189 2013.11.06. Chungnam III
5 (n=20) A. pleuroneumoniae KVCC-BA1400190 2013.11.11. Chungbuk III
A. pleuroneumoniae KVCC-BA1400191 2013.11.13. Chungnam III
A. pleuroneumoniae KVCC-BA1400192 2013.11.18. Gyeonggi III
A. pleuroneumoniae KVCC-BA1400194 2013.12.27. Chungbuk III
A. pleuroneumoniae KVCC-BA1400195 2012.01.02. Chungbuk III
A. pleuroneumoniae KVCC-BA1400216 2012.03.07. Gyeongbuk III
A. pleuroneumoniae KVCC-BA1400217 2012.03.08. Chungnam III
A. pleuroneumoniae KVCC-BA1400219 2012.04.06. Chungnam III
A. pleuroneumoniae KVCC-BA1400220 2013.04.20. Gyeongbuk III
A. pleuroneumoniae KVCC-BA1400226 2013.05.25. Gyeongnam III
7 (n=1) A. pleuroneumoniae KVCC-BA1400212 2012.01.06. Gyeongnam −*
12 (n=1) A. pleuroneumoniae KVCC-BA1400179 2013.02.28. Chungnam −
A. pleuroneumoniae KVCC-BA1400197 2012.01.02. Chungnam I
A. pleuroneumoniae KVCC-BA1400211 2012.01.02. Chungnam I
A. pleuroneumoniae KVCC-BA1400215 2012.03.02. − I
A. pleuroneumoniae KVCC-BA1400218 2012.04.04. Gyeongbuk I
A. pleuroneumoniae KVCC-BA1400224 2012.05.04. Gyeonggi I
A. pleuroneumoniae KVCC-BA1400225 2012.05.24. Gyeongbuk I
Unclassified (n=13) A. pleuroneumoniae KVCC-BA1400227 2012.05.25. Chungnam I
A. pleuroneumoniae KVCC-BA1400228 2012.06.11. Chungbuk −
A. pleuroneumoniae KVCC-BA1400166 2012.08.24. Chungnam I
A. pleuroneumoniae KVCC-BA1400168 2012.09.17. Gyeongbuk −
A. pleuroneumoniae KVCC-BA1400177 2013.02.15. Gyeongbuk −
A. pleuroneumoniae KVCC-BA1400180 2013.03.08. Gyeongnam III
A. pleuroneumoniae KVCC-BA1400183 2013.03.18. Gyeonggi I

* − No detection.

Table 3.
Numbers of A. pleuropneumoniae isolates from pigs between 2012 to 2013
Isolation yr
 Total
2012 2013
No. of pig 177 266 443
No. of A. pleuropneumoniae 35 19 54
 Serotype 1 12 3 15
 Serotype 2 2 2 4
 Serotype 3 0 0 0
 Serotype 5 10 10 20
 Serotype 6 0 0 0
 Serotype 7 1 0 1
 Serotype 8 0 0 0
 Serotype 12 0 1 1
 Unclassified 10 3 13
Table 4.
MIC90 and antimicrobial resistance rates of A. pleuroneumoniae (Serotype 1 and 5)
Antimicrobial agents Breakpoint Serotype 1 (n=15)
 Serotype 5 (n=20)
MIC90 Resistance (%) MIC90 Resistance (%)
Ampicillin −* ≥16 − ≤0.25 −
Penicillin −* ≥8 − 0.25 −
Ceftiofur ≥8 ≤0.25 0.00% ≤0.25 0.00%
Danofloxacin ≥0.25 1 80.00% 1 50.00%
Enrofloxacin ≥2 0.5 0.00% 0.5 0.00%
Spectinomycin −* 64 − 64 −
Gentamicin ≥16 16 60.00% ≥16 20.00%
Neomycin −* ≥32 − ≥32 −
Tilmicosin ≥32 8 0.00% 8 0.00%
Tulathromycin ≥64 32 6.70% 16 0.00%
Florfenicol ≥8 ≥8 67.0% 8 10.0%
Tiamulin ≥32 16 13.3% 16 0.00%
Chlortetracycline ≥16 4 0.00% 4 0.00%

* There is no antimicrobial breakpoint in the Clinical Laboratory Standard Institute (CLSI) guideline.

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