• Journal of Dairy Science and Biotechnology
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• 제36권4호
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• pp.208-219
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• 2018

낙농업 및 유가공 제품의 생산과 유통에서 살모넬라 감염에 의한 살모넬라증의 발생은 빈번하며, 이 세균의 항미생물 제제에 대한 내성 증가 현상 또한 지속되고 있어 새로운 항미생물 제제의 수요는 감소하지 않는다. 세균막의 훼손은 세균 생존을 쉽게 위협할 수 있기 때문에 개발되는 항미생물 제제들은 주로 세균의 막을 표적으로 삼지만, 개발되는 제제들이 실제로 세균막의 훼손을 초래하는지 구별하는 것은 많은 노력과 비용을 수반한다. 본 연구에서는 E. coli 세포막 스트레스에 의해 발현이 유도되고, 세균막 외부공간에서만 위치하며, 그 구조상 많은 단백질의 구조 안정화에 기여할 것으로 예상되는 chaperone 단백질 Spy(spheroplast protein Y)의 유전자에 상응하는 살모넬라 spy 유전자에 gfp(green fluorescence protein) 오페론 융합체를 제조하여, 이 융합체가 Salmonella enterica의 두 혈청형 Enteritidis와 Gallinarum의 세포막 스트레스를 인지하여 GFP 발현량이 크게 증가하는 것을 확인하였다. 또한 세균막 스트레스 신호를 특이적으로 인지하는 이인자 신호전달 체계(two component signal transduction system)인 Bae와 Cpx들이 두 살모넬라 혈청형의 spy 유전자 전사 유도에 필수적임을 확인하였다. 따라서 본 연구에서 사용한 spy-gfp 오페론 융합체는 S. Enteritidis와 S. Gallinarum의 세포막 훼손을 특이적이고 신속하게 인식하는 biosensor로서 활용될 수 있을 것으로 판단된다.

• 한국응용곤충학회지
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• 제41권4호
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• pp.279-284
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• 2002

누에(Bombyx mori) 5령 유충에서 무름병 증상이 발견되었다. 세균성 병원균이 감염 누에 혈액에서 분리되었고, 본 연구에서 동정되었다. 분리된 세균을 각각의 누에 5령 유충에 $5{\times}10^{6}$ cfu(colony-forming unit) 농도로 혈강 주사되었을 때, 뚜렷한 무름병 증상을 유발시켰다. 이렇게 감염된 유충들은 접종 6일 후부터 죽기 시작하여, 10일 경과 후 처리된 유충 모두 사망하였다. Bergey의 세균동정집에 기록된 형태적 및 생리적 형질을 기준으로 이 곤충병원세균이 Staphylo-coccue gallinarum으로 판명되었다. 이 판정은 탄소원 분석 미생물동정장치($MicroLog^{\circledR}$) 분석 결과와 16S-23S rDNA internal transcribed space 구조에 의해서 확인되었다. 이 세균이 발휘하는 살충기작의 일환으로서, 감염 후 누에 유충 혈구에 세포치사 효과를 주어 패혈증을 유발하는 것으로 분석되었다.

• 대한수의학회지
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• 제42권4호
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• pp.513-522
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• 2002

Antigenic ompC genes of S. gallinarum, S. pullorum and S. dublin were characterized among Salmonella spp. isolated from chickens and other animals to identify genetic variation. Salmonella ompC gene fragment (1,027 bp) was amplified by PCR and the amplicons were cloned for comparison of nucleotide sequences. The identity of the sequences between S. gallinarum and S. pullorum, S. gallinarum and S. dublin, S. pullorum and S. dublin was 99.8%, 97.6% and 97.8%, respectively. Also, we found that ompC has some diversity between S. gallinarum and S. pullorum, and other Salmonella spp. which may be useful to type the organisms. Similar to diagnosis in other organisms, the TaqMan PCR method can be applied to rapid and accurate diagnosis of salmonellosis in chickens and other animals. We designed PCR primers and TaqMan probe for flagellin gene (fliC) for detection of Salmonella spp. by TaqMan PCR. The TaqMan PCR method was 10,000 times more sensitive than conventional PCR.

• Journal of Microbiology and Biotechnology
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• 제23권10호
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• pp.1478-1483
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• 2013

In this study the isolation and characterization of three bacteriophages (ST4, L13, and SG3) infecting Salmonella gallinarum were carried out. They were further tested for their in vivo efficacy in phage therapy. All three phages belong to the Siphoviridae family with isometric heads and non-contractile tails. They have a broad host range among serovars of Salmonella enterica. The burst sizes were observed to be 1670, 80, and 28 for ST4, L13, and SG3, respectively. The in vivo efficacy of the phages was tested in chickens. Layer chickens were challenged with S. gallinarum, whereas contact chickens were cohabited without direct challenge. Each bacteriophage was orally inoculated in the form of feed additives. Mortality was observed and S. gallinarum was periodically re-isolated from the livers, spleens, and cecums of the chickens. Bacterial re-isolation from the organs and mortality decreased significantly in both challenged and contact chickens treated with the bacteriophages compared with untreated chickens serving as the control. The three bacteriophages may be effective alternatives to antibiotics for the control of fowl typhoid disease in chickens.

• 한국동물위생학회지
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• 제28권1호
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• pp.1-21
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• 2005

Pullorum disease and Fowl typhoid are kind of poultry specific disease for poultry. The peculiar character of these poultry specific diseases is that it can be infected by transmitting vertically and horizontally, also it is hard to be discovered by clinical sign, and pathology or immunology. So, to develop the PCR method which distinguishes these two genetically similar diseases of separated the specific DNA fragment from each strain and use it for differential diagnosis by subtraction PCR method. Standard strain of S gallinarum and S pullorum, and field isolation strain were verified by biochemistry, It confirmed existence of plasmid by using the PFGE. Then, Isolated DNA from it and used it as materials for the experiment. After cutting genomic DNA of two strains by using Sau 3Al, It ligated primer to tester DNA for PCR amplification and separated specific DNA fragment bacteria with method of subtraction PCR. And, It confirmed that it is a piece of unique DNA in every bacteria using base sequence of separated DNA fragment. 1. The six specific DNA fragment were separated from the DNA of S gallinarum and S pullorum by the subtraction PCR method. 2. In the result of comparison after setting base sequence of each fragment, each separated base sequence of DNA fragment they did not correspond to each other 3. As the result of each DNA fragment is derived from the each strain of DNA, and there was no homology of genomic DNA level in mutual. 4. The fragment originated in plasmid and includes S pullorum did not separate. 5. In the result of searching base sequence in Genebank, it partially shows homology in Salmonella enterica, S typhimurium, S dublin, Escherichia coli, Shigella flexneri, Yersinia pestis, Klebsiella pneumoniae. 6. Primer design by S gallinarum DNA 2, 3 fragment used PCR, They are positive reaction in only S gallinarum at 276, 367 bp position.

• 대한수의학회지
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• 제37권3호
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• pp.569-576
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• 1997

Fowl typhoid(FT) caused by Salmonella gallinarum is an infectious, egg-transmitted disease and characterized by swollen bronze liver, greenish-yellow diarrhea and high mortality in growing and adult chickens. Since 1992 the outbreak of FT has been increased. Several problems have been occurred such as absence of appropriate vaccines and lack of useful therapeutic methods. In these studies we investigated the pathogenicity of S gallinarum isolated in chickens. To compare the pathogenicity among the species of chickens, all chickens were challenged intramuscularly or orally with $1{\times}10^7$ CFU of S gallinarum. The brown-colored layers were more susceptible and white leghone chickens were more resistant than other species. In the brown layer chickens orally challenged, lethal doses ($LD_{50}$) of the isolates were inoculated at 1 day, 2 weeks, 4 weeks and 8 weeks old chickens with amount of $10^{4.2}$, $10^{4.7}$, $10^{7.0}$ and $10^{7.6}$ CFU, respectively. The chickens which were intramuscularly challenged with the less amount than $10^2$ CFU showed higher mortality than that of the chickens orally inoculated with same dose. Also, we investigated the recovery rates of bacteria from various organs of survival chickens which were challenged orally with $5{\times}l0^7$ CFU of S gallinarum. The bacteria was more frequently and isolated earlier from the liver and spleen than from any other ogans. In the pathogenicity test, the white-leghorn chickens which were known as resistant-strain against Salmonella were artificially immunosuppressed using bursectomy and/or dexamethasone treatment. Mortality of chickens with both bursectomized and treated with dexamethasone was higher(90%) than that of the control group(10%), the bursectomized chickens(10%) and the dexamethasone only treated group(20%). It was suggested that the protective mechanism in chickens against S gallinarum may be required both the functions of B-cells and T-cells.

• 대한수의학회지
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• 제37권3호
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• pp.555-568
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• 1997

Fowl typhoid caused by Salmonella gallinarum has increased dramatically since 1992 and has caused a great economic losses in chicken industry by characterizing with high mortality. In these studies, we investigated the immunogenicity and protectivity in chickens which were immunized with outer membrane protein(OMP) extracted from isolates of S gallinarum against challenge with live microorganism. Outer membrane proteins were composed of various sizes of molecular weight including 14K, 22K, 31K, 36K, 40K and 55K and the most of them responded strongly against rabbit antisera in immunoblot analysis. The chickens vaccinated with OMP or vaccinated with whole-cell combined with OMP($200{\mu}g$/chickens) complex showed higher delayed type hypersensitivity(DTH) response than that of whole-cell vaccinated group. The protective rates of OMP or whole-cell combined with OMP complex group against challenge of S gallinarum were higher (above 75%) than those (45~50%) of whole-cell vaccinated group. All vaccines were safe and the body weight-gains of all vaccinated groups were not significantly different (p<0.05) from those of nonvaccinated control group. In vitro tests, OMP stimulated both the proliferation of lymphocytes and T-lymphocytes, and OMP-induced lymphocyte proliferation was higher in the cells of the immunized chickens with OMP than in those from the control chickens.

• 한국가금학회지
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• 제35권1호
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• pp.9-13
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• 2008

양계 산업에서 살모넬라에 의한 질병들 중 가장 중요한 원인체는 S. Gallinarum, S. Purollum, S. Enteritidis로 간주된다. 생화학적 검사에 의한 직접적인 균 분리.동정과 같은 이들 질병에 대한 종래의 진단법은 많은 시간이 소요되며, 특이성 또한 낮다. 본 연구는 3종의 살모넬라균에 의해 야기되는 이들 질병에 대한 빠르고 정확한 진단을 위한 효율적인 진단법에 초점을 두었다. 먼저 종래의 생화학적 검사를 대신하여 새로 고안된 ITSF/ITSR PCR primer를 이용하여 살모넬라균임을 확인하였으며, 증폭된 phase 1 flagellin (fliC) 유전자를 BpmI 또는 BfaI 제한 효소로 처리하여 S. Gallinarum, S. Purollum, S. Enteritidis를 상호 용이하게 감별하였다. 이상의 결과는 3종의 살모넬라균을 효율적으로 진단하기 위해 신속하게 검출하고 감별할 수 있는 유용한 진단법임을 알 수 있었다.

• 대한수의학회지
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• 제44권1호
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• pp.73-82
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• 2004

Chitosan is similar in structure to cellulose and are the second most abundant polysaccharides in nature, comprising the horny substance in the exoskeletons of crabs, shrimp and insects as well as fungi. This study was conducted to access the effect of immunomodulation responses of chitosan(N-acetyl-${\beta}$-D-glucosamine) chicken infected with in Fowl typhoid(Salmonella gallinarum). One-day-old broiler chicks were divided into eight groups: The 1st group was inoculated intra-peritoneally with chitosan and challenged intra-peritoneally with S. gallinarum. The 2nd group was inoculated intra-peritoneally with chitosan. The 3rd group was feeding with chitosan and intra-peritoneally inoculated with cyclophosphamide and challenged intra-peritoneally S. gallinarum. The 4th group was feeding with chitosan and intra-peritoneally with cyclophosphamide. The 5th group was feeding with chitosan and challenged intra-peritoneally with S. gallinarum. The 6th group was feeding with chitosan. The 7th group was challenged intra-peritoneally with S. gallinarum. The 8th group was nontreated-uninfected control group. The results shows that $CD4^+$, $CD8^+$ and B lymphocyte in lymphoid organs of chickens treated with chitosan increased in especially $CD4^+$, $CD8^+$ lymphocytes (p<0.05). The group of feeding chitosan showed the significantly increased $CD4^+$, $CD8^+$ and B lymphocyte than inoculated intra-peritoneally with chitosan. As the result suggests that the feeding of chitosan induced immunostimulatant effect than the inoculation intra-peritoeally of chitosan.

• Applied Biological Chemistry
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• 제47권1호
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• pp.27-32
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• 2004

가금티프스는 닭, 오리, 칠면조, 꿩 등 가금류에 Salmonella gallinarum이 원인균이 되어 발병하는 질병이다. 본 연구는 가금티프스를 방제하기 위한 생균제 개발을 위한 기초조사의 목적으로 가금티프스 원인균인 5. gallinarum 의 생육을 저해시킬 수 있는 길항균을 야생꿩에서 분리, 선발했다. 최종적으로 3종의 우수 길항균주를 선발하였으며 선발, 분리된 길항균을 동정한 결과 Ll9, L33균주는 similarity가 100%인 Sphinromonas sanguis로 동정 되었고 L50균주는 similarity가 96.2%로써 Sphinromonas sanguis의 근연종으로 동정되었다. 선발된 길항균주가 생산한 길항물질의 특성을 확인해본 결과, S. gallinarum을 저해하는 물질이 고분자성 물질이고 내열성임을 추정 할 수가 있었다. S. sanguis Ll9의 경우 배양 20시간 이후부터 길항물질이 생산되었으며 탄소원, 질소원을 각각 maltose, $(NH_4)_2SO_4$를 첨가하였을 때 가장 높은 길항력을 나타내었다. S. sanguis L33의 경우 배양 22시간 이후 부터 길항물질이 생산되었으며 탄소원, 질소원을 각각 galactose, urea를 첨가하였을 때 가장 높은 길항력을 나타내었다. S. sanguis L50은 배양 10시간 이후부터 길항물질을 생산하였으며 탄소원, 질소원을 각각 saccharose, $(NH_4)_2S_2O_8$음로 하였을 때 가장 높은 길항력을 나타내었다.