• Title/Summary/Keyword: Salmonella typhimurium

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Characterization of Drug-Resistant Salmonella enterica Serotype Typhimurium by Antibiograms, Plasmids, Integrons, Resistance Genes, and PFGE

  • Benacer, Douadi;Thong, Kwai Lin;Watanabe, Haruo;Puthucheary, Savithri Devi
    • Journal of Microbiology and Biotechnology
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    • v.20 no.6
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    • pp.1042-1052
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    • 2010
  • Forty-seven Salmonella Typhimurium (33 zoonotic, 14 clinical) strains were tested for antimicrobial resistance using the standard disk diffusion method. The presence of relevant resistance genes and class 1 integrons were investigated by using PCR. Pulsed-field gel electrophoresis (PFGE) and plasmid profiling were carried out to determine the genomic diversity of Salmonella Typhimurium. Approximately 57.4% of the S. Typhimurium strains were multidrug resistant (MDR) and showed high resistance rates to tetracycline (70.2%), sulfonamides (57.4%), streptomycin (53.1%), ampicillin (29.7%), nalidixic acid (27.6%), kanamycin (23.4%), chloramphenicol (21.2%), and trimethoprim (19.1%). Resistance towards cephalosporins was noted for cephalothin (27.6%), cephradine (21.2%), amoxicillin clavulanic acid (17.0%), and cephalexin (17.0%). Resistance genes, $bla_{TEM}$, strA, aadA, sul1, sul2, tetA, tetB, and tetC, were detected among the drug-resistant strains. Thirtythree strains (70.2%) carried class 1 integrons, which were grouped in 9 different profiles. DNA sequencing identified sat, aadA, pse-1, and dfrA genes in variable regions on class 1 integrons. Thirty-five strains (74.4%) were subtyped to 22 different plasmid profiles, each with 1-6 plasmids (2.0 to 95 kb). PFGE subtyped the 47 strains into 39 profiles. In conclusion, high rates of multidrug resistance were found among the Malaysian Salmonella Typhimurium strains. The emergence of multidrug-resistant Salmonella Typhimurium to cephalosporin antibiotics was also observed. The strains were very diverse and no persistent clone was observed. The emergence of MDR Salmonella Typhimurium is a worldwide problem, and this report provides information for the better understanding of the prevalence and epidemiology of MDR S. Typhimurium in Malaysia.

Anaerobic Acid Tolerance Response in Salmonella typhimurium (Salmonella typhimurium의 혐기적 산내성도 평가)

  • Kim, Young-Chan;Lee, Sun;Lee, Kyung-Mi;Im, Sung-Young;Park, Yong-Geun;Baek, Hyung-Seok;Park, Kyung-Ryang;Lee, In-Soo
    • Journal of Life Science
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    • v.9 no.2
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    • pp.169-175
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    • 1999
  • Salmonella typhimurium can encounter a wide variety of environments during its life cycle. In nature, S. typhimurium can experience and survive dramatic acid stresses that occur in diverse ecological niches ranging from pond water to phagolysosomes. These survival mechanism is aquired by the Acid Tolerance Response(ATR) in Salmonella. The ATR of S. typhimurium is a complex inducible phenomenon in which exposures to slight or moderate low pH will produce a stress response capable of protecting the organism against more severe acid challenges. ATR in Salmonella has two different systems that are called RpoS dependent and independent. We found that ATR in anaerobic was showed RpoS independent because rpoS$\Omega$AP had ATR as S. typhimurium UK1. Using the P22 MudJ(Km, lacZ) operon fusion technique and a lethal selection procedure combining low pH(pH4.5) and sodium acetate(10mM, pH4.5), we isolated LF487 aatA::MudJ which showed acid sensitive in anaerobic condition. aatA locus was determined at 12 min on Salmonella Genetic Map. The survival rate of aatA mutant was showed significantly diminished at pH4.3 than virulent wild type Salmonella in anaerobic condition(5% $CO_2$, 5% H$_2$, 90% $N_2$). Therefore isolated gene was confirmed important gene for anaerobic ATR system.

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Direct and Quantitative Analysis of Salmonella enterica Serovar Typhimurium Using Real-Time PCR from Artificially Contaminated Chicken Meat

  • Park, Hee-Jin;Kim, Hyun-Joong;Park, Si-Hong;Shin, Eun-Gyeong;Kim, Jae-Hwan;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1453-1458
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    • 2008
  • For quantitative PCR assay of Salmonella enterica serovar Typhimurium in food samples, a real-time PCR method was developed, based on DNA genome equivalent. Specific primers and probe designed based on the STM4497 gene of S. Typhimurium LT2 showed the specificity to S. Typhimurium. Threshold cycle (Ct) values of real-time PCR were obtained from a quantitative standard curve with genomic DNA of Salmonella Typhimurium. In addition, the recovery of S. Typhimurium inoculated artificially to chicken samples with $4.5{\times}10^5$ to 4.5 CFU/ml was evaluated by using real-time PCR and plate-count methods. Result showed that the number of cells calculated from the real-time PCR method had good correlation with that of the plate-count method. This real-time PCR method could be applicable to the detection and quantification of S. Typhimurium in food samples.

Genomic Approaches for Understanding the Characteristics of Salmonella enterica subsp. enterica Serovar Typhimurium ST1120, Isolated from Swine Feces in Korea

  • Kim, Seongok;Kim, Eunsuk;Park, Soyeon;Hahn, Tae-Wook;Yoon, Hyunjin
    • Journal of Microbiology and Biotechnology
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    • v.27 no.11
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    • pp.1983-1993
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    • 2017
  • Salmonella enterica subsp. enterica serovar Typhimurium, one of the most common foodborne pathogens, is transmitted mainly through contaminated food derived from infected animals. In this study, S. Typhimurium ST1120, an isolate from pig feces in Korea, was subjected to whole-genome analysis to understand its genomic features associated with virulence. The genome of ST1120 was found to have a circular chromosome of 4,855,001 bp (GC content 52.2%) and a plasmid of 6,863 bp (GC content 46.0%). This chromosome was predicted to have 4,558 open reading frames (ORFs), 17 pseudogenes, 22 rRNA genes, and 86 tRNA genes. Its plasmid was predicted to have three ORFs. Comparative genome analysis revealed that ST1120 was phylogenetically close to S. Typhimurium U288, a critical isolate in piggery farms and food chains in Europe. In silico functional analysis predicted that the ST1120 genome harbored multiple genes associated with virulence and stress resistance, including Salmonella pathogenicity islands (SPIs containing SPI-1 to SPI-5, SPI-13, and SPI-14), C63PI locus, ST104 prophage locus, and various antibiotic resistance genes. In accordance with these analysis results, ST1120 showed competence in invasion and survival abilities when it was added to host cells. It also exhibited robust resistance against antibiotics in comparison with other S. Typhimurium strains. This is the first report of the complete genome sequence of S. Typhimurium isolated from swine in Korea. Comparative genome analysis between ST1120 and other Salmonella strains would provide fruitful information toward understanding Salmonella host specificity and developing control measures against S. Typhimurium infection.

Comparison of tdcA Expression Between Escherichia coli and Salmonella enterica Serovar Typhimurium

  • Kim, Min-Jeong;Lim, Sang-Yong;Ryu, Sang-Ryeol
    • Journal of Microbiology and Biotechnology
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    • v.21 no.3
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    • pp.252-255
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    • 2011
  • Both Escherichia coli (E. coli) and Salmonella enterica serovar Typhimurium (S. Typhimurium) have a tdc operon that encodes enzymes involved in a metabolic pathway for the degradation of L-serine and L-threonine. However, S. Typhimurium does not have the tdcR gene, which is a positive regulator in E. coli. In the present study, transcriptional analysis revealed that tdcA expression in E. coli is higher under anaerobic than aerobic growth conditions, but the opposite is true in S. Typhimurium. Interestingly, a tdcR mutant strain of E. coli showed a similar expression pattern to that observed in S. Typhimurium and was also induced by anaerobic shock. These results suggest that the induction of tdcA expression by anaerobic conditions is observable when tdcA expression is low owing to the absence of TdcR.

Selective Detection of Salmonella sp. and Salmonella Typhimurium in Meat by Polymerase Chain Reaction (PCR을 이용한 육류 내 Salmonella sp. 및 Salmonella Typhimurium 분리 검출)

  • Joo, Jong-Won;Hong, Kyung-Pyo;Kim, Yong-Hui;Cho, Sang-Buem
    • 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.

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젖당과 사멸 살모넬라 함유 사료가 Salmonella typhimurium 인공 감염 육계 병아리의 생산성과 면역 반응에 미치는 영향

  • Choe, Jun-Yeong;Im, Jin-Taek;Park, In-Gyeong;Choe, Do-Yeol;Lee, Hye-Jeong;Lee, Beom-Gyu;Go, Tae-Song
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2005.11a
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    • pp.78-79
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    • 2005
  • Effects of dietary lactose or killed Salmonella on the performance, immune response and anti-oxidant system was studied in chicks innoculated with Salmonella typhimurium. In 27 days of age broiler, dietary lactose decreased performance, while dietary lactose and killed Salmonella elevated plasma peroxidase activity and IL-1 level in supernatant of PBMC stimulated with LPS. When broiler chicks innoculated with Salmonella, performance, activities of erythrocyte MnSOD and plasma peroxidase were enhanced after 7 days of the innoculation. Dietary lactose and killed Salmonella increased activity of erythrocyte MnSOD, plasma peroxidase, proliferation of PBMC stimulated with LPS and IL-1 level in the supernatant after 15 days of the innoculation. The result indicated that dietary lactose and killed Salmonella have modulated innate immune response and antioxidant system in broiler chicks innoculated with Salmonella typhimurium.

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Protective effect of bacteriophages against Salmonella Typhimurium infection in weaned piglets (이유자돈에서 Salmonella Typhimurium 감염에 대한 박테리오파지의 방어 효능)

  • Kim, Sung-Jae;Kim, Jae-Hoon;Jun, Soo-Yeon;Paik, Hyoung Rok;Han, Jeong-Hee
    • Korean Journal of Veterinary Service
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    • v.37 no.1
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    • pp.35-43
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    • 2014
  • Salmonellosis has caused heavy losses in swine industry and implications for public health. Recently, the urgent problem of antibiotic resistance due to multidrug-resistant Salmonella spp. has been on the rise. The use of host-specific bateriophages as a biocontrol is one possible alternative. In this study, clinical signs, growth performance, quantification and detection of antigen, histopathological changes of gastrointestinal tracts were analyzed comparatively in weaned piglets according to administration of bacteriophages and challenge with Salmonella (S.) Typhimurium. Piglets challenged with S. Typhimurium after administered with bacteriophages showed reduced clinical signs, higher growth performance, lower bacterial shedding, lower quantificational value of antigens in intestines, higher V/C ratio and higher the number of goblet cells in intestines than piglets administered without bacteriophage and challenged with S. Typhimurium. These results indicate that feeding contained with bacteriophages has effect to prevent infection of S. Typhimurium in weaned piglets and suggest that a use of bacteriophage can be considered a valid antibiotic alternative.

Mutagenic Activity of Smoke Flavoring Processed from Oak and Apple Wood on Manufacturing Temperature (굴참나무와 사과나무로부터 제조한 훈연액의 제조온도에 따른 돌연변이원성에 관한 연구)

  • 강희곤;이경호;홍희선;박상진;김창한
    • Food Science of Animal Resources
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    • v.18 no.3
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    • pp.203-208
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    • 1998
  • The study was carried out to screen mutagenicity of smoking materials for the determination of optimum smoking temperature for meat products. Wood materials employed for smoking were oak and apple trees. Temperatures of the generator for manufacturing of smoke flavoring were set to 250$^{\circ}C$, 400$^{\circ}C$ and 500$^{\circ}C$, respectively. Mutagenic activities of smoke flavoring were assayed according to Ames test using Salmonella typhimurium TA98 and TA 100. In oak wood smoke flavoring, Salmonella typhimurium TA98 without S-9 mix showed strong mutagenic activities at the concentration of 6$\mu\textrm{g}$/plate(250$^{\circ}C$), 4$\mu\textrm{g}$/plate(400$^{circ}C$) and 6$\mu\textrm{g}$/plate(500$^{\circ}C$). Salmonella typhimurium TA100 with S-9 mix showed strong mutagenic activities at the concentration of 10$\mu\textrm{g}$/plate(250$^{\circ}C$), 20$\mu\textrm{g}$/plate(400$^{\circ}C$) and 10$\mu\textrm{g}$/plate(500$^{\circ}C$). Salmonella typhimurium TA98 with S-9 mix showed strong mutagenic activities at the concentration of 30$\mu\textrm{g}$/plate(250$^{\circ}C$), 40$\mu\textrm{g}$/plate(400$^{\circ}C$) and 20$\mu\textrm{g}$/plate(500$^{\circ}C$). Salmonella typhimurium TA100 with S-9 mix showed strong mutagenic activities at the concentration of 30$\mu\textrm{g}$/plate(250$^{\circ}C$), 50$\mu\textrm{g}$/plate(400$^{\circ}C$) and 20$\mu\textrm{g}$/plate(500$^{\circ}C$). Salmonella typhimurium TA100 without S-9 mix showed strong mutagenic activities at the concentration of 10$\mu\textrm{g}$/plate(250$^{\circ}C$), 20$\mu\textrm{g}$/plate(400$^{\circ}C$) and 20$\mu\textrm{g}$/plate(500$^{\circ}C$). Salmonella typhimurium TA98 with S-9 mix showed strong mutagenic activities at the concentration of 30$\mu\textrm{g}$/plate(250$^{\circ}C$), 40$\mu\textrm{g}$/plate(400$^{\circ}C$) and30$\mu\textrm{g}$/plate(500$^{\circ}C$). Salmonella typhimurium TA100 with S-9 mix showed strong mutagenic activities at the concentrations 30$\mu\textrm{g}$/plate(500$^{\circ}C$). Salmonella typhimurium TA100 with S-9 mix showed strong mutagenic activities at the concentration of 30$\mu\textrm{g}$/plate(250$^{\circ}C$), 20$\mu\textrm{g}$/plate(400$^{\circ}C$) and 30$\mu\textrm{g}$/plate(500$^{\circ}C$). From these results, it could be concluded that optimum smoking temperature for meat products should be set below 400$^{\circ}C$, that the compounds like benzo[a]pyrene etc. contain a variety of mutagenic potentials, which could be generated at the higher smoking temperature.

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Induced Autophagy Regulates Salmonella enterica serovar Typhimurium Infection in Murine Macrophage (쥐의 큰포식세포주에서 자가포식현상에 의한 Salmonella enterica serovar Typhimurium의 감염 조절)

  • Lee, Sunhye;Kim, Ju-Young;Lee, Hyo-Ji;Jung, Yu-Jin
    • 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.