• 한국수의병리학회지
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• 제7권1호
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• pp.27-41
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• 2003

The purpose of this study was to evaluate the effect of a subsequent infection of porcine reproductive and respiratory syndrome(PRRS) virus to pigs with A. pleuropneumonia in pigs. Twenty three 7-weeks-old commercial pigs were infected with PRRS virus and/or A. pleuropneumoniae serotype 5 intratracheally. Feed conversion, clincal signs, gross and histopathological lesions and immunohistochemical findings were examined. 1. Feed conversion ratio in dual-infected pigs with PRRS virus and A. pleuropneumoniae were higher than that of single- infected pigs with PRRS virus or A. pleuropneumoniae. 2. Dual-infected pigs with PRRS virus followed by A. pleuropneumoniae showed more severe clinical signs and gross, histopathological and immunohistochemical pulmonary lesions. The results indicated that dual infections with PRRS virus and A. pleuropneumoniae caused more severe respiratory lesions and growth retardation in pigs than single infection with PRRS virus or A. pleuropneumoniae.

• 한국임상수의학회지
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• 제14권1호
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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.

• Journal of Microbiology and Biotechnology
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• 제20권4호
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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.

• Journal of Microbiology and Biotechnology
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• 제25권10호
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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.

• 대한수의학회지
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• 제52권3호
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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.

• 대한수의학회지
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• 제43권2호
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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.

• 한국동물위생학회지
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• 제30권1호
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• pp.51-66
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• 2007

The purpose of this study was to evaluate the effect of a subsequent infection of porcine reproductive and respiratory syndrome(PRRS) virus to pigs with A pleuropneumonia. Twenty three 7-week-old commercial pigs were infected intratracheally with PRRS virus and/or A pleuropneumoniae serotype 5. Serum antibody titers were examined by an enzyme-linked immunosorbent assay(ELISA) and proportion of porcine leukocyte subpopulations in peripheral blood was examined by flow cytometry. In this experiment, antibodies against PRRS virus and A pleuropneumoniae were detected at 2 weeks and 1 week postinfection and the number of antibody positive pigs were gradually increased. And in proportion to leukocyte subpopulations in peripheral blood of pigs infected with A pleuropneumoniae compared with pigs administrated with saline, the proportion of PoCD4 and N cells were increased(P<0.1). Furthermore, in proportion to leukocyte subpopulations in peripheral blood of pigs infected with PRRS virus followed by A pleuropneumoniae compared with pigs administrated with saline, the proportion of MHC class II, PoCD4 and B cells were significantly increased(P<0.1). The results indicated that dual infection with PRRS virus and A pleuropneumoniae induced the stronger immune responses associated with macrophages and Th cells in pigs than single infection with PRRS virus or A pleuropneumoniae.

• 대한수의학회지
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• 제36권1호
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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.

• 대한수의학회지
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• 제45권2호
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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.

• 한국동물위생학회지
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• 제25권3호
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• pp.237-243
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• 2002

The study was performed to investigate the antibody distributions of 4 swine respiratory disease including M hyopneumoniae, P multocida, A pleuropneumoniae serotype 2 and 5 in Daegu area by enzyme-linked immunosorbent assay(ELISA). 1. The overall sero-positive rates were 55.6% in June, 48.0% in August, 51.3% in October and 25.4% in November. 2. The positive reaction rates to M hyopneumoniae, P multocida, A pleuropneumoniae serotype 2 and 5 were found to be 50.0%, 36.5%, 55.0%, and 42.0% respectively. 3. The antibody titers were distributed on range 20～80 in M hyopneumoniae, 20～80 in P multocida, 160～640, 20～80 in A pleuropneumoniae serotype 2 and 5.