• Title, Summary, Keyword: Actinobacillus pleuropneumoniae

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Serotyping of Actinobacillus pleuropneumoniae by Coagglutination Test (Coagglutination 반응법에 의한 Actinobacillus pleuropneumoniae의 혈청형 조사)

  • 예재길
    • Journal of Veterinary Clinics
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    • v.14 no.1
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    • pp.37-41
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    • 1997
  • For the inspection of the occurrence situation of porcine pleuropneumonia and serotyping of Actinobacillus pleuropneumoniae strains isolated from lung lesions of pig in Korea, a series of experimentation have been carried out by the isolation and identification of A pleuropneumoniae, serotyping by coagglutination test, observation of lung lesion and clinical signs from 360 cases of porcine pneumonia in Clinical Pathology Laboratory, Bayer Veterinary Medical Research Institute. The results could be summarized as follows. The reaction of coagglutination between the reference antigens and the specific reagents of A pleuropneumoniae was strongly agglutinatied within 30 seconds without cross reaction. The 89 strains of A pleuropneumoniae were isolated from 360 cases of porcine pleuropneumonis and the biochemical properties of the isolates were same as the reference strains. The 89 isolated strains could be serotyped 39 strains as setotype 5, 34 strains as serotype 2, 8 strains as serotype 3, 2 strains as serotype 7 by coagglutination test, respectively. The clinical signs of pleuropneumonia were weakness, fever, anorexia, dyspnea and laboured breath in the later stages. The gross lesions of lung were haemorrhages, enlargement of interlobular septa, nodular formation and adhesion of the pleura.

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Serotype and antimicrobial susceptibility of Actinobacillus pleuropneumoniae isolates from pigs in Korea (돼지에서 분리한 Actinobacillus pleuropneumoniae의 혈청형 분포 및 항생제 감수성)

  • Jung, Ji-Youl;Jang, Hyun
    • Korean Journal of Veterinary Research
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    • v.52 no.3
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    • pp.177-181
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    • 2012
  • Actinobacillus (A.) pleuropneumoniae is the causative agent of pleuropneumonia which is one of the most important respiratory diseases in pigs worldwide. A total of 32 A. pleuropneumoniae isolates from diseased pigs during 2008 to 2010 were serotyped by polymerase chain reaction method. The susceptibility of the isolates to 13 antimicrobial agents were determined by disk diffusion test. In all the 32 isolates examined in this study, serotype 5 (16 isolates: 50%), 1 (7 isolates: 21.9%), 2 (5 isolates: 15.6%) and 12 (1 isolate: 3.1%) were found. Of all tested antimicrobial agents, resistance to oxytetracycline was found in 96.9% of isolates, followed by resistance to amikacin (81.2%), neomycin (68.7%), kanamycin (53.1%), penicillin (50.0%), gentamicin (43.7%), florfenicol (25.0%), ampicillin (18.7%), colistin (9.4%), trimethoprim/sulfamethoxazole, ceftiofur (8.3%), amoxicillin/clavulanic acid (3.1%) and enrofloxacin (0%). Oxytetracycline or florfenicol-resistant isolates were examined for the presence of resistance gene. Among the 31 oxytetracycline-resistant isolates, tetB, tetH and tetO genes were detected in 22 (71%), 8 (26%) and 1 (3%) isolates, respectively. The floR genes were detected in 8 (100%) of the 8 florfenicol-resistant A. pleuropneumoniae isolates.

Antibacterial Compound against Pasturella multiocida and Actinobacillus pleuropneumoniae Causing Porcine Pneumonia

  • Lyoo, Young-Soo;Park, Dong-Ki;Lee, Sang-Mok;Choi, Yi-Don;Jung, Ji-Hyun;Jun, Tae-Il;Ahn, Hong-Suk;Lee, Chul-Hoon;Lim, Yoong-Ho
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.350-353
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    • 2001
  • Porcine pneumonia is caused by Pasturella multiocida and Actinobacillus pleuropneumoniae. To identify a potent drug for antipneumonia therapy, several herbal compounds showing antibacterial effects were screened, and it was found that a methanol extract of Coptidis rhizoma root stem exhibited activity against both pneumonia-causing bacteria. Using an activity-guided fragmentation procedure, an isoquinoline alkaloid was isolated which would be responsible for the antibacterial activities against P. multocida and A. pleuropneumoniae.

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Biochemical characteristics and serotypes of Actinobacillus pleuropneumoniae isolated from pneumonic lungs of pigs (돼지 폐렴병소에서 분리한 Actinobacillus pleuropneumoniae의 특성에 관한 연구)

  • Jung, Byeong-yeal;Cho, Gil-jae;Kim, Bong-hwan;Cho, Kwang-hyun
    • Korean Journal of Veterinary Research
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    • v.36 no.1
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    • pp.181-186
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    • 1996
  • The present study was conducted to investigate the biochemical and serologic characteristic of Actinobacillus pleuropneumoniae isolated from pneumonic lungs of pigs during the period from January 1992 to April 1993. A pleuropneumoniae was isolated from 17(27.0%) of 63 growing pigs with respiratory signs and 21(6.4%) of 330 pneumonic lungs of slaughtered pigs. The seasonal isolation frequency of A pleuropneumoniae was higher in winter and spring than that in summer or fall. The biochemical and cultural properties of A pleluropneumoniae isolated from the pneumonic lungs of pigs were identical to those of the reference strains used. The isolates were highly susceptible to ampicillin, cephalothin, ceftiofur, ciprofloxacin(MIC : ${\leq}0.39{\mu}g/ml$) and moderately susceptible to amikacin, chloramphenicol, erythromycin, kanamycin, methicillin, penicillin-G, streptomycin(MIC : 0.78~25IU or ${\mu}g/ml$), respectively. Sulfadimethoxine, sulfamerazine, tylosine showed no response to the isolates(MIC : ${\geq}100{\mu}g/ml$). Among the 38 isolates, 21(55.3%) and 13(34.2%) were resistant to oxytetracycline aid lincomydn, respectively(MIC : ${\geq}50IU$ or ${\mu}g/ml$). The majority of 38 A pleuropneumoniae isolates were turned out as serotype 2(47.4%) or serotype 5(54.7%) and the remaining 3 isolates were evenly classified to serotype 7, 10 or 12. It was noted A pleuropneumonine serotype 5 isolates were more resistant to oxytetracycline than serotype 2 isolates.

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Mass expression of Apx I and Apx II of Actinobacillus pleuropneumoniae in Escherichia coli (대장균에서 흉막폐렴균 독소 Apx I과 Apx II의 대량발현)

  • Kim, Tae-Jung;Lee, Bong-Joo;Lee, Jae-Il
    • Korean Journal of Veterinary Research
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    • v.45 no.2
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    • pp.185-189
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    • 2005
  • Actinobacillus pleuropneumoniae is the causative agent of a porcine contagious pleuropneumonia. Among several virulence factors including exotoxin (Apx toxins), LPS, transferrin-binding proteins, OMPs, and some proteases, Apx toxins have been major targets for the protection study. In this study, cloning and expression of A. pleuropneumoniae Apx I and Apx II toxin, which are produced by all highly virulent strains, were performed by Escherichia coli expression system. Genes coding Apx I and II toxin were amplified from the A. pleuropneumoniae serotype 5 genomic DNA using polymerase chain reaction and cloned to a prokaryotic expression vector, pRSET. Expression of the Apx I and Apx II coding sequences in E. coli resulted in the formation of insoluble inclusion bodies purified according to a denaturing purification protocol, which employs the use of guanidium. Recombinant proteins were purified using $Ni^{2+}$-charged resin affinity purification. This expression and purification system made it possible to produce Apx I and Apx II in large amounts for further immunologic studies.

Differential Gene Expression in the Pathogenic Strains of Actinobacillus pleuropneumoniae Serotypes 1 and 3

  • Xie, Fang;Zhang, Mingjun;Li, Shuqing;Du, Chongtao;Sun, Changjiang;Han, Wenyu;Zhou, Liang;Lei, Liancheng
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.789-797
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    • 2010
  • The limited information on differential gene expression in the different serotypes of Actinobacillus pleuropneumoniae has significantly hampered the research on the pathogenic mechanisms of this organism and the development of multivalent vaccines against A. pleuropneumoniae infection. To compare the gene expressions in the A. pleuropneumoniae strains CVCC259 (serotype 1) and CVCC261 (serotype 3), we screened the differentially expressed genes in the two strains by performing representational difference analysis (RDA). Northern blot analyses were used to confirm the results of RDA. We identified 22 differentially expressed genes in the CVCC259 strain and 20 differentially expressed genes in the CVCC261 strain, and these genes were classified into 11 groups: (1) genes encoding APX toxins; (2) genes encoding transferrin-binding protein; (3) genes involved in lipopolysaccharide (LPS) biosynthesis; (4) genes encoding autotransporter adhesin; (5) genes involved in metabolism; (6) genes involved in the ATP-binding cassette (ABC) transporter system; (7) genes encoding molecular chaperones; (8) genes involved in bacterial transcription and nucleic acid metabolism; (9) a gene encoding protease; (10) genes encoding lipoprotein/membrane protein; and (11) genes encoding various hypothetical proteins. This is the first report on the systematic application of RDA for the analysis of differential gene expression in A. pleuropneumoniae serotypes 1 and 3. The determination of these differentially expressed genes will serve as an indicator for future research on the pathogenic mechanisms of A. pleuropneumoniae and the development of a multivalent vaccine against A. pleuropneumoniae infection.

Identification of Actinobacillus pleuropneumoniae Genes Preferentially Expressed During Infection Using In Vivo-Induced Antigen Technology (IVIAT)

  • Zhang, Fei;Zhang, Yangyi;Wen, Xintian;Huang, Xiaobo;Wen, Yiping;Wu, Rui;Yan, Qigui;Huang, Yong;Ma, Xiaoping;Zhao, Qin;Cao, Sanjie
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1606-1613
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    • 2015
  • Porcine pleuropneumonia is an infectious disease caused by Actinobacillus pleuropneumoniae. The identification of A. pleuropneumoniae genes, specially expressed in vivo, is a useful tool to reveal the mechanism of infection. IVIAT was used in this work to identify antigens expressed in vivo during A. pleuropneumoniae infection, using sera from individuals with chronic porcine pleuropneumonia. Sequencing of DNA inserts from positive clones showed 11 open reading frames with high homology to A. pleuropneumoniae genes. Based on sequence analysis, proteins encoded by these genes were involved in metabolism, replication, transcription regulation, and signal transduction. Moreover, three function-unknown proteins were also indentified in this work. Expression analysis using quantitative real-time PCR showed that most of the genes tested were up-regulated in vivo relative to their expression levels in vitro. IVI (in vivo-induced) genes that were amplified by PCR in different A. pleuropneumoniae strains showed that these genes could be detected in almost all of the strains. It is demonstrated that the identified IVI antigen may have important roles in the infection of A. pleuropneumoniae.

Cloning, Sequencing and Expression of apxIA, IIA, IIIA of Actinobacillus pleuropneumoniae Isolated in Korea (국내 분리 흉막폐렴균의 apxIA, IIA, IIIA 유전자 Cloning, 염기서열 분석 및 단백질 발현)

  • Shin, Sung-jae;Cho, Young-wook;Yoo, Han-sang
    • Korean Journal of Veterinary Research
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    • v.43 no.2
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    • pp.247-253
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    • 2003
  • Actinobacillus pleuropneumoniae causes a highly contagious pleuropneumoniae in swine. The bacterium produces several virulence factors such as exotoxin, LPS, capsular polysaccharide, etc. Among them, the exotoxin, called Apx, has been focused as the major virulence factor, and the toxin consists of 4 gene cluster. apx CABD. apxA is the structural gene of toxin and has four different types, I, II, III, and IV. As the first step of development of a new subunit vaccine, the three different types of apxA gene were amplified from A. pleuropneumoniae isolated from Korea by PCR with primer designed based on the N- and C-terminal of the toxin. The sizes of apxIA, IIA and IIIA were 3,073, 2,971 and 3,159bps, respectively. The comparison of whole DNA sequences of apxIA, IIA and IIIA genes with those of the reference strain demonstrated 98%, 99% and 98% homology, respectively. In addition, the phylogenetic analysis was performed based on the amino acid sequences compared with 12 different RTX toxin family using the neighbor-joining method. ApxA proteins of Korean isolates were identical with reference strains in this study. All ApxA proteins were expressed in E. coli with pQE expression vector and identified using Western blot with polyclonal antibodies against culture supernatants of A. pleuropneumoniae serotype 2 or 5. The sizes of each expressed ApxA protein were about 120, 110, 125 kDa (M.W.), respectively. The results obtained in this study could be used for the future study to develop a new vaccine to porcine pleuropneumoniae.

Isolation, identification and serological investigation of Actinobacillus pleuropneumoniae in slaughtered pigs (도축돈에서의 Actinobacillus pleuropneumoniae 분리, 동정 및 감염률 조사)

  • Kim, Kyung-Eon;Ku, Kyung-Nyer;Ko, Jae-Hyung;Moon, Hyeong-Jun;Choi, Kwon-Rag;Song, Eun-Ah;Park, Mi-Young
    • Korean Journal of Veterinary Service
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    • v.36 no.3
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    • pp.181-186
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    • 2013
  • This study was conducted to isolate the Actinobacillus pleuropneumoniae (APP) and to find out the distribution of 15 serovars mainly in southern Gyeonggi province, Korea. From July 2011 to Nov. 2012, a total of 2,204 slaughter pigs (110 herds) were inspected for evaluation of APP like pneumonic lesions. 48 (33.8%) APP strains were isolated from the 142 lungs and identified using PCR assays (cps, apx/omlA, biovar). Consequently, the serotype ratio were as in the following; type2 41.7% (n=20), type5 33.3% (n=16), type12 10.4% (n=5), type1 6.2% (n=3), type4 and 7 2.1% (n=1) and unknown 4.2% (n=2). Also serological test was implemented for 452 (83 herds) serum samples randomly collected from above slaughter pigs using commercial ELISA kits. The positive ratio of each serotype for tested pigs were 19.1% (77/404) on [2], 7.1% (32/452) on [3, 6, 8], 6.9% (28/404) on [5a, 5b], 6.2% (28/452) on [4, 7], 2.8% (9/320) on [12], 2.0% (9/452) on [1, 9, 11] and 0.0% (0/452) on [10]. And 49.3% (223/452) of pigs were positive on apxIV antibody. On the basis of latter screening test, the infected farm ratio accounted for 71.1% (59/83) and that was much higher than previously reported data.

Expression of the Apx Toxins of Actinobacillus pleuropneumoniae in Saccharomyces cerevisiae and Its Induction of Immune Response in Mice

  • Park Seung-Moon;Choi Eun-Jin;Kwon Tae-Ho;Jang Yong-Suk;Yoo Han-Sang;Choi Woo Bong;Park Bong-Kyun;Kim Dae-Hyuk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.362-366
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    • 2005
  • Actinobacillus pleuropneumoniae is an important pig pathogen, which is responsible for swine pleuropneumonia, a highly contagious respiratory infection. To develop subunit vaccines for A. pleuropneumoniae infection, the Apx toxin genes, apxI and apxII, which are thought to be important for protective immunity, were expressed in Saccharomyces cerevisiae, and the induction of immune responses in mice was examined. The apxI and apxII genes were placed under the control of a yeast hybrid ADH2-GPD promoter (AG), consisting of alcohol dehydrogenase II (ADH2) and the GPD promoter. Western blot analysis confirmed that both toxins were successfully expressed in the yeast. The ApxIA and ApxIIA-specific IgG antibody response assays showed dose dependent increases in the antigen-specific IgG antibody titers. The challenge test revealed that ninety percent of the mice immunized with ApxIIA or a mixture of ApxIA and ApxIIA, and sixty percent of mice immunized with ApxIA survived, while none of those in the control groups survived longer than 36 h. These results suggest that vaccination of the yeast ex­pressing the ApxI and ApxII antigens is effective for the induction of protective immune responses against A. pleuropneumoniae infections in mice.