• Journal of Dairy Science and Biotechnology
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• v.36 no.4
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• pp.208-219
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• 2018

Emergence of drug resistant strains of Salmonella enterica threatens milk processing and related dairy industries, thereby increasing the need for development of new anti-bacterials. Developments of antibacterial drugs are largely aimed to target the bacterial envelope, but screening their efficacy on bacterial envelope is laborious. This study presents a potential biosensor for envelope-specific stress in which a gfp reporter gene fused to spy gene encoding a periplasmic chaperone protein Spy (spheroplast protein y) that can sense envelope stress signals transduced by two major two-component signal transduction systems BaeSR and CpxAR in Salmonella enterica serovars Enteritidis and S. Gallinarum. Using spy-gfp operon fusions in S. Enterititis and S. Gallinarum, we found that spy transcription in both serovars was greatly induced when Salmonella cells were forming the spheroplast and were treated with ethanol or a membrane-disrupting antibiotic polymyxin B. These envelope stress-specific inductions of spy transcription were abrogated in mutant Salmonella lacking either BaeR or CpxR. Results illustrate that induction of Spy expression can be efficiently triggered by two-component signal transduction systems sensing envelope stress conditions, and thereby suggest that monitoring the spy transcription by spy-gfp operon fusions would be helpful to determine if developing antimicrobials can damage envelopes of S. Enteritidis and S. Gallinarum.

• Korean journal of applied entomology
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• v.41 no.4
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• pp.279-284
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• 2002

Flacherie symptom was found in the fifth instar larvae of silkworm, Bombyx mori. The bacterial pathogen was isolated from the hemolymph of the infected silkworm and identified. The isolated bacteria caused a significant flacherie pathogenicity to the fifth instar larvae of B. mori when $5{\times}10^{6}$ cfu (colony-forming unit) of the bacteria was injected into each larva. The infected larvae began to die at 6 days after injection and resulted in complete mortality at 10 days. The bacterium was identified as Staphylococcus gallinarum based on the morphological and physiological characteristics described in Bergey's manual. This identification was further supported by the characters of carbohydrate utility analyzed from a bacterial identification system ($MicroLog^{\circledR}$) and also by the molecular structure of 165-23S rDNA internal transcribed spaces. As an insecticidal action, S. gallinarum caused hemolymph septicemia by its cytotoxic effect on the hemocytes of B. mori.

• Korean Journal of Veterinary Research
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• v.42 no.4
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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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• v.23 no.10
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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.

• Korean Journal of Veterinary Service
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• v.28 no.1
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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.

• Korean Journal of Veterinary Research
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• v.37 no.3
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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.

• Korean Journal of Veterinary Research
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• v.37 no.3
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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.

• Korean Journal of Poultry Science
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• v.35 no.1
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• pp.9-13
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• 2008

Salmonella enterica serotype gallinarum biovar Gallinarum or Pullorum and Salmonella enterica serotype Enteritidis are the most important diseases in poultry industry. Transitional diagnosis methods of these diseases such as direct isolation and identification by a biochemical test are time consuming with low specificity. In this study, we have focused on the suitable procedure for the rapid and accurate diagnosis of diseases derived from the three Salmonella strains. We initially confirmed Salmonella species by PCR using a specific ITSF/ITSR primer pair instead of biochemical test, and then the PCR-amplified phase 1 flagellin (fliC) using a specific fliCF/fliCR primer pair was digested with a restriction endonuclease, Bpm I and/or Bfa I, to discriminate among S. Pullorum, S. Gallinarum, and S. Enteritidis. We found that these methods could be applied to field isolates of the three Salmonella strains to detect and to discriminate rapidly for convenient diagnosis.

• Korean Journal of Veterinary Research
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• v.44 no.1
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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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• v.47 no.1
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• pp.27-32
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• 2004

The antagonistic microorganisms against Salmonella gallinarum causing fowl typhoid were isolated from the gut of wild pheasant. The isolated L19, L33, L50 strains were showed the characteristics of isolated Gram negative, rods, catalase positive and oxidase negative. Finally, all strains were identified as Sphingomonas sanguis by $Biolog^{\circledR}$ system. The optimal carbon sources of Sphingomonas sanguis L19, L33 and L50 for the these growth ~ere glucose, saccharose, and fructose respectively. But the optimal carbon sources of S. sanguis L19,L33, L50 for the antagonistic material production were maltose, galactose, and saccharose respectively. The optimal nitrogen sources of S. sanguis L19, L33, L50 for the growth were yeast extract, yeast extract, and $NH_4H_2PO_4$ respectively. But the optimal nitrogen sources of S. sanguis L19, L33 and L50 for the antagonistic material production were $(NH_4)_2SO_4$ urea, $(NH_4)_2S_2O_8$ espectively.