• Journal of the East Asian Society of Dietary Life
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• v.19 no.2
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• pp.295-300
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• 2009

The specificity and sensitivity of oligonucleotide primers were examined for the rapid detection of Salmonella in meat samples. The oligonucleotide primers used in this study were designed with the modification of mdh and invA sequence in the chromosome of Salmonella Typhimurium. Through the subsequent analysis of the specificity and sensitivity of the primers, two types of oligonucleotide primers, SLM1 and SLT4 were selected for the detection of Salmonella genus specific and S. Typhimurium species specific, respectively. The lowest detection limit of each primer was represented as 1 cell per reaction when reacted with a prepared DNA solution. The detection efficiency of the two primers was analysed with beef and pork samples intentionally contaminated with a mixture of Salmonella culture, and three preparation methods -, namely direct reaction after extraction, enrichment after extraction, and DNA extraction after enrichment for PCR reaction, - were also compared. No differences were found in the results according to meat sources and preparation methods.

• Journal of Microbiology
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• v.42 no.3
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• pp.216-222
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• 2004

In a previous paper, the ogdH gene that encodes 2-oxoglutarat dehydrogenase was isolated from Salmonella typhimurium. The catalytic N-terminal region in the enzyme was found to be very specific for the Salmonella species. Therefore, the aim of the present study was to detect S. typhimurium in food sources using primers designed for OGDH-l and OGDH-2 which were based on the salmonella-specific region of the ogdH gene. A simple polymerase chain reaction (PCR) detection method was developed to detect low numbers of S. typhimurium in a chicken meat microbial consortium. Using the ogdH-specific primers under stringent amplification conditions and for gene probe analysis, fewer than 100 colony-forming units (CFUs) were detectable when pure cultures were employed. When the PCR assay was run on S. typhimurium-contaminated meat contents, only the positive meat samples containing as few as 200 CFUs reacted to the assay. The method employed for sample processing is simple and it was determined to provide a sensitive means of detecting trace amounts of S. typhimurium-specific sequences in the presence of mixed meat microbial populations. When compared with six representative intestinal gram-negative bacterial strains in foods, including Vibrio parahaemolyticus, V. vulnificus, Enterobacter cloacae, E. coli O157:H7, Pseudomonas aeruginosa, and Proteus sp., S. typhimurium had a unique and distinct PCR product (796 bp). In conclusion, the two OGDH primers were found to be rapid and sensitive detectors of Salmonella spp for the PCR method.

• Korean Journal of Veterinary Research
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• v.43 no.1
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• pp.77-86
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• 2003

Pullorum disease due to Salmonella enterica subspecies enterica bioserovar Pullorum (S. pullorum) is reported to be an endemic disease in domestic poultry flocks. The pulsed-field gel electrophoresis (PFGE) subtyping method was used to assess the extent of genetic diversity and clonality of most of salmonella serotypes and other diverse bacterial species from animals and environmental samples in worldwide. Nowadays, PFGE has already been evaluated as a gold standards for molecular subtyping of salmonella serotypes compared with other molecular analysis methods. PFGE of XbaI digested chromosomal DNA from 23 strains of S. pullorum gave 5 distinctive pulsotypes (from SXPI to SXPV) with 5% confidence range of Dice coefficients, indicating that PFGE is very discriminative and that multiple clones of S. pullorum have been existed and diffused all of domestic poultry flocks industries since 1995. Two dominant pulsogroups (SXA & SXB) appeared as a major clones in this country, because they had consistently been recovered from diverse sources including both chicken organs and raw feed materials between 1995 and 1998. In addition, the matching percentage of PFGE profiles (PFP) among strains from both chickens and feed ingredients provides indirect evidence of the possible transmission of pullorum disease from contaminated raw feed ingredients for chicken production. In calculating of discrimination index (DI) for PFGE method by Simpson's index, DI was appeared as 0.917. Therefore, this index suggested that the present PFGE would seem to be a desirable and confident molecular typing method for S. pullorum strains. To our knowledge for pullorum disease, this is the first study to compare S. pullorum strains from chicken organs and feed samples using the PFGE.

• Food Science and Preservation
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• v.16 no.6
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• pp.965-970
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• 2009

PCR technology has been widely used to detect and quantify microbial pathogens in foodstuffs, because the technique is rapid, sensitive, and selective. In this study, detection of contaminating pathogenic bacteria on shells of chicken eggs was performed using both a commercial multiplex polymerase chain reaction (PCR) kit and a viable count method employing a selective medium. The PCR kit was capable of detecting Campylobacter jejuni, Escherichia coli O157:H7, Staphylococcus aureus, Bacillus cereus, Vibrio parahaemolyticus, Listeria monocytogenes, Yersinia enterocolitica, Salmonella species, and Shigella species. Using the PCR method, five bacterial species were detected from 30 samples (33.3%) of 90 batches of eggs commercially available in a market. PCR products from B. cereus, S. aureus, L. monocytogenes, Y. enterocolitica, and E. coli O157:H7 were detected, and the numbers and frequencies of positive samples were 17 (18.8%), 12 (13.3%), 15 (16.6%), 16 (17.7%),and 4 (4.4%), respectively. None of any Salmonella species, C. jejuni, V. parahaemolyticus, or Shigella species was detected in this study. The results of PCR testing were confirmed using a typical viable count method employing a selective medium. We suggest that the multiplex polymerase chain reaction (mPCR) assay is a rapid and reliable method for detection of pathogenic bacteria contaminating eggs.

• Journal of Biosystems Engineering
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• v.30 no.2 s.109
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• pp.121-126
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• 2005

In this study, a commercial electronic nose system was used to detect contamination of Salmonella bacteria. Odors from growth media contaminated with Salmonella typhimurium, Salmonella enteritidis, or Escherichia coli were collected and analyzed to evaluate a possibility of rapid detection of pathogen. Odor chromatograph showed that S. typhimurium, S. enteritidis, and E. coli had 7,6, and 9 main peaks, respectively. Retention time and intensity of the peaks were distinct for different bacteria species. Principal component analysis (PCA) were also performed to clarify odor differences. Analysis results showed that the odors for uncontaminated growth medium were differently grouped from the odors of contaminated one. The odor from the bacteria growth identified with two principal components, PC 1 and PC2. In PCA figures, odor groups were moved from left to right of PC 1 with elapse of the bacteria growth time. The electronic nose system could detect odors of S. typhimurium, S. enteritidis, E. coli when their concentration were $1.85\times10^6\;cfu/g,\;2.25\times10^6\;cfu/g,\;and\;1.8\times10^5 cfu/g$, respectively.

• Journal of Dairy Science and Biotechnology
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• v.36 no.3
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• pp.146-154
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• 2018

There are more than 25 species of Mentha plants, which are aromatic perennial herbs. Currently, these species are being widely used with great interest because of various clinical findings regarding their health benefits. This is due to the abundance of volatile compounds that could expedite environmental interactions such as protection against herbivores, parasites, pathogens, and so on. Therefore, in this study, the antimicrobial effect of Mentha piperita (peppermint) oil on Bacillus cereus, Staphylococcus aureus, Cronobacter sakazakii, and Salmonella Enteritidis were investigated using the spot-on-lawn method. The results show that Mentha piperita (peppermint) oil exhibited antimicrobial activities against Bacillus cereus, Staphylococcus aureus, and Cronobacter sakazakii; however, it did not inhibit the growth of Salmonella Enteritidis. This shows that the antimicrobial effect of Mentha piperita (peppermint) oil is effective against both Gram-positive and Gram-negative bacteria. Hence, in the present study, Mentha piperita (peppermint) oil was shown to have strong antimicrobial activities; it could be used as a potential food additive for improving the quality of various milk-based products due to its various bioactive properties. Future studies should be conducted for manufacturing functional dairy products with the addition of peppermint oil to prevent and/or alleviate specific diseases.

• International Journal of Industrial Entomology and Biomaterials
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• v.37 no.2
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• pp.95-99
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• 2018

In insect defense system, antimicrobial peptides (AMPs) are one of important biological molecules to survive in a variety of environments. Insect can synthesize AMPs to protect against invading pathogens in humoral immune response. Taking more advantage of biological antimicrobial molecules, we report antibacterial activity of two cecropin type peptides, cecropin and moricin, isolated from the silkworm against four salmonella species. In this work, we purified antimicrobial candidate peptides (AMCP) from the extracts of immune challenged silkworm larval hemolymph by two-step chromatographic purification procedure, cation exchange and gel permeation chromatography. The molecular weights of purified peptides were estimated to be about 4 ~ 5 kDa by Tricin SDS-PAGE analysis, and identified as silkworm cecropin and moricin by NCBI BLAST homology search with their N-terminal amino acid sequences. As antibacterial activity assay, the purified peptides showed stronger antibacterial activity against Salmonella pathogens with an MIC value of $1{\sim}4{\mu}g/mL$. Therefore two cecropin type peptides purified from the silkworm will be valuable potential materials for development of new natural antibiotics.

• Molecules and Cells
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• v.43 no.12
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• pp.989-1001
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• 2020

Salmonella enterica serovar Typhimurium (S. Typhimurium) is a facultative intracellular pathogen that causes salmonellosis and mortality worldwide. S. Typhimurium infects macrophages and survives within phagosomes by avoiding the phagosome-lysosome fusion system. Phagosomes sequentially acquire different Rab GTPases during maturation and eventually fuse with acidic lysosomes. Lysophosphatidylcholine (LPC) is a bioactive lipid that is associated with the generation of chemoattractants and reactive oxygen species (ROS). In our previous study, LPC controlled the intracellular growth of Mycobacterium tuberculosis by promoting phagosome maturation. In this study, to verify whether LPC enhances phagosome maturation and regulates the intracellular growth of S. Typhimurium, macrophages were infected with S. Typhimurium. LPC decreased the intracellular bacterial burden, but it did not induce cytotoxicity in S. Typhimurium-infected cells. In addition, combined administration of LPC and antibiotic significantly reduced the bacterial burden in the spleen and the liver. The ratios of the colocalization of intracellular S. Typhimurium with phagosome maturation markers, such as early endosome antigen 1 (EEA1) and lysosome-associated membrane protein 1 (LAMP-1), were significantly increased in LPC-treated cells. The expression level of cleaved cathepsin D was rapidly increased in LPC-treated cells during S. Typhimurium infection. Treatment with LPC enhanced ROS production, but it did not affect nitric oxide production in S. Typhimurium-infected cells. LPC also rapidly triggered the phosphorylation of IκBα during S. Typhimurium infection. These results suggest that LPC can improve phagosome maturation via ROS-induced activation of NF-κB pathway and thus may be developed as a therapeutic agent to control S. Typhimurium growth.

• Korean Journal of Microbiology
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• v.50 no.1
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• pp.27-32
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• 2014

Autophagy is one of the lysosomal degradation pathways to maintain cellular homeostasis. The damaged proteins or organelles are uptaken through extra- and intra-cellular stress, starvation and infected pathogens, subsequently, autophagosomes are fused with lysosomes to break down the molecules. Salmonella enterica serovar Typhimurium (S. Typhimurium), intracellular bacteria, cause acute gastroenteritis and food poisoning. Given that autophagy induced by S. Typhimurium plays an important role in the cells to control the infection, we identify whether the induction of autophagy with rapamycin, chemical inducer of autophagy, before infection regulates S. Typhimurium infection. After treatment of rapamycin or 3-methyladenine (3-MA), autophagy inhibitor, RAW264.7 cells were infected with S. Typhimurium. Pretretment of rapamycin decreased the growth rate of S. Typhimurium in the cells; otherwise, pretreatment of 3-MA increased the growth rate of S. Typhimurium. The expression of autophagy-related genes was significantly increased in the S. Typhimurium-infected cells pretreated with rapamycin. To examine whether induced autophagy by rapamycin control the infection with increase the production of reactive oxygen species (ROS) and nitric oxide (NO), antibacterial radical substrates were measured in infected cells followed by the treatment with either rapamycin or 3-MA. NO production increased in RAW264.7 cells; otherwise, ROS production remained unchanged during the infection. These findings suggest that inducing autophagy with rapamycin reveals antimicrobial activity as producing NO against S. Typhimurium infection in mouse macrophages.

• 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.