• Journal of Microbiology
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• 제44권3호
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• pp.320-326
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• 2006

Pasteurella multocida is an important veterinary and opportunistic human pathogen. In particular, strains of P. multocida serogroup D cause progressive atrophic rhinitis, and produce a potent, intracellular, mitogenic toxin known as P. multocida toxin (PMT), which is encoded by the toxA gene. To further investigate the toxigenic and pathogenic effects of PMT, a toxA-deleted mutant was developed by homologous gene recombination. When administrated to mice, the toxigenicity of the toxA mutant P. multocida was drastically reduced, suggesting that the PMT constributes the major part of the toxigenicity of P, multocida. Similar results were obtained in a subsequent experiment, while high mortalities were observed when toxA(+) P. multocida bacterial culture or culture Iysate were administrated. Mice immunized with toxA(-) P. multocida were not protected (none survived) following challenge with toxA(+) P. multocida or bacterial culture Iysate (toxin). These results suggest that the toxigenicity of P. multocida is mainly derived from PMT.

• 대한수의학회지
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• 제47권1호
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• pp.59-65
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• 2007

Pasteurella multocida Pasteurella multocida toxin (PMT) is a causal pathogenin atrophic rhinitis in pigs. To investigate the protective immunity and vaccination effect of recombinantPMT, the gene for PMT was isolated from the infective P. multocida D:4. The 2.3 kb XhoI/PstI fragment(PMT2.3) of PMT gene was expressed in E. coli BL21 (DE3) using the induced expression vector system.The recombinant protein of PMT2.3 having molecular weight of 84 kDa was purified by Ni-afinitycolumn chromatography. The PMT2.3 raised slightly less anti-PMT antibody titer than formalin-killedwhole cel, however, it showed more protective imunity against P. multocida D:4 infection in vaccinationand chalenge.

• Journal of Microbiology and Biotechnology
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• 제20권12호
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• pp.1756-1763
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• 2010

Pasteurella multocida serogroup D strain, which produces P. multocida toxin (PMT), is a widespread and harmful pathogen of respiratory diseases such as pneumonia and progressive atrophic rhinitis (PAR) in swine. Vaccination has been considered the most desirable and effective approach for controlling the diseases caused by toxigenic P. multocida. To investigate the antigenicity and immunogenicity of partial fragments of recombinant PMT, recombinant proteins of the N-terminal (PMT-A), middle (PMT-B), C-terminal (PMT-C), and middle-C-terminal (PMT2.3) regions of PMT were successfully produced in an Escherichia coli expression system. The molecular masses of PMT-A, PMT-B, PMT-C, and PMT2.3 were ca. 53, 55, 35, and 84 kDa, respectively, purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. All the recombinant proteins except for PMT-A showed immune responses to antisera obtained from a swine showing symptoms of PAR. Moreover, high titers of PMT-specific antibodies were raised from mice immunized with each of the recombinant proteins; however, the immunoreactivities of the antibodies to authentic PMT and heat-inactivated whole bacteria were different, respectively. In the protection study, the highest protection against homologous challenge was shown in the case of PMT2.3; relatively poor protections occurred for the other PMT fragments.

• Journal of Microbiology
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• 제45권4호
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• pp.364-366
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• 2007

The immunological role of the Pasteurella multocida toxin (PMT) in mice was examined using a PMT mutant strain. After a nasal inoculation, the mutant strain failed to induce interstitial pneumonia. Moreover, PMT had no significant effect on the populations of CD4+, CD8+, CD3+, and CD19+ immunocytes in blood or on the populations of CD4+ and CD8+ splenocytes (P<0.01). However, there was a significant increase in the total number of cells in the BAL samples obtained from the wild-type P. multocida-inoculated mice. On the other hand, the level of IL-l expression decreased when the macrophages from the bronchio-alveolar lavage were stimulated with PMT. Overall, PMT appears to play some role (stimulating and/or inhibiting) in the immunological responses but further studies will be required to confirm this.

• 대한수의학회지
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• 제40권1호
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• pp.56-62
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• 2000

Pasteurella multocida is kind of commensal bacteria in the upper respiratory tract of pigs. It is classified toxigenic and nontoxigenic strains based on the production of dermonecrotic toxin. Toxigenic strain is most associated with atrophic rhinitis which brings great economical loss in swine industry. However, toxigenic and nontoxigenic strains do not differ by diagnostic biochemical reaction or morphology. One of recently developed techniques, PCR detects the toxigenic P multocida. Amplification of an 846-nucleotide fragment of toxA gene was developed. The fragment amplified by PCR was detected in P multocida type D not type A. The PCR amplification was as sensitive as it could detect 1 pg of P multocida DNA. We compared the result of the PCR with the enzyme linked immunosorbent assay (ELISA) in a test for 40 swine nasal swabs. All of these isolates were toxin negative based on the ELISA while 2 isolates were detected in the PCR technique. in addition to accuracy, as required for rapid detection from contaminated nasal swabs, toxigenic P multocida was recovered efficiently from contaminated culture without inhibition of the PCR. The results show that the PCR detection of toxigenic P multocida directly form nasal swabs are feasible.

• 대한수의학회지
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• 제48권1호
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• pp.53-60
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• 2008

We have investigated efficiency of a recombinant subunit Pasteurella multocida toxin (PMT) that was mixed with a vaccine consisted of inactivated whole cells of Bordetella bronchiseptica, P. multocida (types A and D). For verification of the efficacy of the vaccine, all experimental pigs (suckling piglets, sow and gilts) in the three farms were vaccinated. Antibody titers against B. bronchiseptica and P. multocida type A of the vaccinated pigs by microplate agglutination were significantly higher than those of the control pigs (p < 0.05). Similar patterns were observed in the analysis of anti- PMT neutralizing antibody by serum neutralizing method using Vero cell (p < 0.05). Anti- P. multocida type D antibody titer of the vaccinated sows and gilts by ELISA showed significant differences with those of the non-vaccinated pigs (p < 0.05). Although antibody titers increased, it was unable to find out the difference in the clinical signs between the vaccinated and non-vaccinated pigs. However, the increase in body weight of the vaccinated piglets was observed in comparison with the non-vaccinated piglets on a farm. At slaughtering of the pigs, pathological lesions in the turbinate bones of the vaccinated pigs were significantly lower than those of the non-vaccinated pigs (p < 0.001). These results suggested that efficacy of the vaccine in pigs demonstrated to protect against atrophic rhinitis in Korea.

• 한국동물위생학회지
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• 제33권2호
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• pp.173-176
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• 2010

Here, we report the suitability of using a resin-type chitosan formulae as a sustained-releasing form adjuvant in comparison with commercially well-known Freund's adjuvants. To induce the immunological responses, N-terminal region of Pasteurella multocida toxin was used as an antigen, which was found to be protective immunogen against P. multocida infection. Mice immunized with chitosan resin formulae showed statistically significant antibody induction (P<0.001) as much as that of Freund’s adjuvants. As a result, the resin-type sustained-releasing form chitosan formulae is thought to be a good candidate for a new type adjuvant.

• 대한수의학회지
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• 제34권3호
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• pp.511-516
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• 1994

이 실험은 도축돈의 폐에서 Pasteurella multocida 감염의 발생빈도를 조사하고 분리된 Pasteurella multocida의 항생제에 대한 약제감수성, 생화학적특성, 협막혈청형의 분류 및 독소생성능에 대해서 조사하였다. 실험재료로는 영남지방의 돼지 450두가 사용되었으며, 재료채취는 1992년 4월부터 1993년 3월 사이에 시행되었다. P multocida는 도축돈의 폐 450예에서 80주가 분리되어 17.7%의 분리율을 나타내었다. 분리균 대부분의 생화학적 및 배양성상은 reference strain의 것과 일치하였으며, 분리균 80주의 협막혈청형을 동정한 결과 77주가 type A, 나머지 3주가 type D로 나타나 각각 96.3%, 3.8%의 분포비를 나타내었다. 항생제에 대한 감수성검사에서 모든 분리균주는 ampicillin, ceftiofur, cephalothin, ciprofloxacin, penicillin-G 등에 대해서는 매우 감수성이 높았으며 이들 분리균중 일부는 sulfadimethoxine과 streptomycin에 내성을 나타내었다. 분리균 80주중 61주가 독소생성능이 있는 것으로 나타났으며, type A 77, type D 3주중 76.6%가 각각 독소생성능이 있는 것으로 나타나 협막혈청형간에 독소생성능의 뚜렷한 차이가 인정되지 않았다.

• 한국동물위생학회지
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• 제19권2호
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• pp.126-138
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• 1996

Lungs from 109 slaughter pigs with gross lesions indicating enzootic Pneumonia of pigs(EPP) and 16 grossly normal lungs, all originating from seven different herds, were subjected to microbiological examinations. The microbiological studies were included both bacterial and mycoplasmal culture. From lungs of 125 slaughter pigs, 87.2% pigs were pneumonia lesions alone or complexly Mycoplasma spp., pasteurella multocidu(p. multocida), Streptococcus spp., and Actinobacillus pleuropneumoniue(A. pleuropneumoniae) were detected in 39.4%, 42.2%, 13.8%, and 3.7% of the pneumonic lungs, respectively. P. multocida was the most frequently isolated organism in pneumonic lungs. Mycoplasmas not isolated organism in 33.9% the pneumonic lungs even If [here are gross lessions mycoplasmas. The amounts of pneumonia in lungs with Mycoplasma spp. alone, a concurrence of Mycoplasma spp. and P. multocida, p. multocida alone, a concurrence of P. multocida and A. pleuropneumoniae, and a concurrence of Mycoplasma spp. and A pleurdpneumoniae were 10.1%, 22.7%, 18.7%, 25%, and 30%, respectively These findings indicated that p. multocida might be involved in the pathogenesis of pneumonia in slaughter pigs. Mycoplasma spp. was also, in this study, associated with higher frequency of pneumonia. The frequency of pigs snout lesion grade 0∼5 inclusive were 27.2%, 28%, 19.2%, 16%, 6.4%, and 3.2% from 125 slaughter pigs. 32(25.6%) pigs were positive and 13~30% in the pigs from seven herds were found to be infected with atrophic rhintis (AR). A total of 46 P. multocida strains In pneumonic lungs were further characterized by capsular serotyping and testing for production of dermonecrotic toxin. 42(91.3%) of strains were capsular A and 4(8.7%) were type D. Out of the type A and type D strains, 86% and 75% were toxigenic, respectively.