• Title/Summary/Keyword: Salmonella assay

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Mutagenicity of N-Nitrosodimethylamine in Salmonella / Microsome Assay and the Effect of Vitamin C on the Formation of N-Nitrosodimethylamine (Salmonella / Microsome Assay 에서의 N-nitrosodimethylamine의 돌연변이 유발성과 N-nitrosodimethylamine의 생성에 대한 비타민 C의 영향)

  • 김소희;박건영;서명자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.3
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    • pp.260-265
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    • 1991
  • The effective method to detect the mutagenicity of N-nitrosodimethylamineI (NDMA) by using Salmonella/microsome assay was studied. The Effect of vitamin C on the mutagenicity of the formed NDMA and during the formation of NDMA from nitrite and secondary amine was also investigated. Aroclor 1254-induced hamster S9 mix was more effective in activation NDMA than rat S9 mix induced by Aroclor 1254 or phenobarbital. Dimethyl sulfoxide and ethanol suppressed the mutagenic effect of NDMA, however, phosphate butter (pH 7.4), distilled water, 95% methanol and Tween 80 + water (1 : 4) were the appropriate dissolving system in the mutagenicity test of NDMA. Vitamin C did not show any inhibitory effect on the mutagenicity of the formed NDMA. However, the revertants of Salmonella typhinutrium TA 100 were significntly reduced (p<0.05) when vitamin C was added to the reaction mixture of nitrite and dimethylamine during the formation of NDMA. The amount of the formed NDMA was analyzed using HPLC and the level was decreased by about 95%. Thus it was concluded that vitamin C inhibited greatly the formation of NDMA from nitrite and dimethylamine.

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Establishment and Application of Polymerase Spiral Reaction Amplification for Salmonella Detection in Food

  • Xu, Wenli;Gao, Jun;Zheng, Haoyue;Yuan, Chaowen;Hou, Jinlong;Zhang, Liguo;Wang, Guoqing
    • Journal of Microbiology and Biotechnology
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    • v.29 no.10
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    • pp.1543-1552
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    • 2019
  • Salmonella is a common zoonotic and foodborne pathogen that causes high morbidity and mortality in developing countries. In this study, we established and validated a polymerase spiral reaction (PSR) assay which targeted the conserved invasion gene (invA) of Salmonella by SYBR Green I indicator methods. Subsequently, assays for determination of the optimal conditions for optimal specificity and sensitivity of PSR were performed. We performed comprehensive evaluations using loop-mediated isothermal amplification (LAMP) and real-time PCR. A total number of 532 samples of daily food were analyzed by PSR. Twenty-seven bacterial strains were tested in the specificity assay, from which positive results were obtained only for 14-Salmonella strains. However, none of the 13 non-Salmonella strains was amplified. Similarly with LAMP and real-time PCR, the detection limit of the PSR assay was 50 CFU/ml. The PSR method was also successfully applied to evaluate the contamination with Salmonella in 532 samples of daily food, corroborating traditional culture method data. The novel PSR method is simple, sensitive, and rapid and provides new insights into the prevention and detection of foodborne diseases.

Study on Mutagenicity of DehydroevodiamineㆍHCl(DHED) (치료제 DehydroevodiamineㆍHCl(DHED)의 변이원성 연구)

  • 성이숙;정성윤;정주연;채규영;진미령;최봉웅;장병모;김대경
    • YAKHAK HOEJI
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    • v.46 no.3
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    • pp.208-212
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    • 2002
  • Dehydroevodiamine HCl (DHED), which is a component separated from Evodia rutaecarpa Bentham, has novel anticholinesterase and antiamnesic activities in the scopolamine-induced amnesia model. Several studies suggest that DHED might be an effective drug for the Alzheimer's disease and the vascular type of dementia. In order to evaluate the mutagenic potential of DHED, Salmonella typhimurium reversion assay, chromosomal aberration test on Chinese hamster lung cells, in vivo micronucleus assay using mouse bone marrow cells, and comet assay were performed. DHED did not increase the number of revertant in the reverse mutation test using Salmonella typhimurium TA1535, TA1537, TA98, TA100. DHED HCl, at concentration of 5 and 10 $\mu\textrm{g}$/mι, increased the number of chromosome aberrated Chinese hamster lung cells with 5 and 10%, respectively. In mouse micronucleus test, no significant increase in the occurrence of micronucleated polychromatic erythrocyte was observed in ICR mice orally administered with DHED. DHED was tested for ability to induce genotoxic effect in L5178Y cells (mouse lymphoma cells) using the single cell gel electrophoresis assay (comet assay). In comet assay, tail moment did not increase in L5178Y cells treated with 10, 100, 300 $\mu$M DHED.

Direct Detection of Escherichia coli, Staphylococcus aureus, and Salmonella spp. in Animal-derived Foods Using a Magnetic Bead-based Immunoassay

  • Kim, Jong-Hui;Yoo, Jae Gyu;Ham, Jun-Sang;Oh, Mi-Hwa
    • Food Science of Animal Resources
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    • v.38 no.4
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    • pp.727-736
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    • 2018
  • In this study, an immuno-magnetic bead (IMB)-based assay was developed to simultaneously detect Escherichia coli, Staphylococcus aureus, and Salmonella spp. and was tested in four animal-derived foods: beef, ham, egg, and ricotta cheese. The IMB-based assay exhibited good specificity by binding to five E. coli serotypes [capture efficiency (CE) average (avg.) 90.4%], five S. aureus strains (CE avg. 91.4%), and five Salmonella serotypes (CE avg. 95.4%) but not binding to non-target bacteria (CE<10%). Furthermore, the assay detected all three pathogens with a detection limit of 10 CFU/g without the need for enrichment or additional platforms. Since the results demonstrated that the IMB-based assay can effectively separate and enrich target bacteria from a variety of animal-derived food matrixes, the assay exhibits good specificity for potential use in providing rapid, immunological, presumptive identification of pathogenic bacteria.

Effects of Sulforaphane, Grapefruit Seed Extracts, and Reuterin on Virulence Gene Expression Using hilA and invF Fusion Strains of Salmonella typhimurium

  • Kim, Ji-Yeun;Ryu, Sang-Ryul;Ji, Geun-Eog
    • Food Science and Biotechnology
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    • v.16 no.5
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    • pp.778-782
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    • 2007
  • This study assessed the effects of the antimicrobial substances sulforaphane, grapefruit seed extracts (GSE), and reuterin on the expression of Salmonella HilA and InvF virulence gene using a LacZY assay (${\beta}$-galactosidase assay) with hilA:lacZY and invF:lacZY fusion strains of Salmonella typhimurium SL1344. Salmonella was grown for 8 hr at $37^{\circ}C$ in the presence of diluted antimicrobial substances ($2\;{\mu}g/mL$ sulforaphane, $20\{\mu}g/mL$ GSE, and 0.26 mM reuterin) at concentrations that did not inhibit the cellular growth of Salmonella. Sulforaphane inhibited the expression of HilA and InvF by 50-90 and 20-80%, respectively. GSE also inhibited the expression of both genes, but to a lesser degree. Among the 3 antimicrobial substances, reuterin showed the least inhibition, which was abolished after 3-4 hr. None of the antimicrobial substances inhibited the ${\beta}$-galactosidase enzyme activity of S. typhimurium. The assay used in this study represents a very sensitive method for screening bioactive substances that inhibit the expression of virulence genes in Salmonella.

Development of Liposome Immunoassay for Salmonella spp. using Immunomagnetic Separation and Immunoliposome

  • Shin, Jung-Hee;Kim, Myung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.18 no.10
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    • pp.1689-1694
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    • 2008
  • The ability to detect Salmonella spp. is essential in the prevention of foodborne illness. This study examined a Salmonella spp. detection method involving the application of immunomagnetic separation and immunoliposomes (IMS/IL) encapsulating sulforhodamine B (SRB), a fluorescent dye. A quantitative assay was conducted by measuring the fluorescence intensity of SRB that was produced from an immunomagnetic bead-Salmonella spp.-immunoliposome complex. The results indicated detection limits of $2.7{\times}10^{5}$ and $5.2{\times}10^{3}$ CFU/ml for Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) and Salmonella enterka subsp. enterka serovar Typhimurium (S. Typhimurium), respectivley. The signal/noise ratio was improved by using 4% skim milk as a wash solution rather than 2% BSA. In addition, higher fluorescence intensity was obtained by increasing the liposome size. Compared with the conventional plating method, which takes 3-4 days for the isolation and identification of Salmonella spp., the total assay time of to h only including 6 h of culture enrichment was necessary for the Salmonella detection by IMS/IL. These results indicate that the IMS/ IL has great potential as an alternative rapid method for Salmonella detection.

Comparison of Gold Biosensor Combined with Light Microscope Imaging System with ELISA for Detecting Salmonella in Chicken after Exposure to Simulated Chilling Condition

  • Mi-Kyung Park
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.228-234
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    • 2023
  • In this study, the performance of a gold biosensor combined with light microscope imaging system (GB-LMIS) was comparatively evaluated against enzyme-linked immunosorbent assay (ELISA) for detecting Salmonella under simulated chilling condition. The optimum concentration of antiSalmonella polyclonal antibodies (pAbs) was determined to be 12.5 and 100 ㎍/ml for ELISA and GBLMIS, respectively. GB-LMIS exhibited a sufficient and competitive specificity toward three tested Salmonella among only. To mimic a real-world situation, chicken was inoculated with Salmonella cocktail and stored under chilling condition for 48 h. The overall growth of Salmonella under chilling condition was significantly lower than that under non-exposure to the chilling condition (p < 0.05). No significant differences in bacterial growth were observed between brain heart infusion and brilliant green broth during the enrichment period (p > 0.05). Finally, both GB-LMIS and ELISA were employed to detect Salmonella at every 2-h interval. GB-LMIS detected Salmonella with a competitive specificity by the direct observation of bacteria on the sensor using a charge-coupled device camera within a detection time of ~2.5 h. GB-LMIS is a feasible, novel, and rapid method for detecting Salmonella in poultry facilities.

Inhibitory Effect of Main Pine Needle Extracts on the Chemically Induced Mutagenicity (주요 솔잎 추출물의 돌연변이 억제효과)

  • Kim, Eun-Jeong;Jung, Sung-Won;Choi, Keun-Pyo;Ham, Seung-Shi;Kang, Ha-Young
    • Korean Journal of Food Science and Technology
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    • v.30 no.2
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    • pp.450-455
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    • 1998
  • Pine has been known as a traditional medicinal plant and as showing a physically beneficial function to a human being. Therefore, this study was performed to investigate the physiological activities of main pine neddles. Ethanol extracts from pinus needles did net exhibit any mutagenicity. On the contrary, inhibitory effects of ethanol extract were observed on mutagenicity induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), 4-nitroquinoline-1-oxide (4NQO), 3-amino-1,4-dimethyl-5H-pyrido-(4,3-b)indol (Trp-P-1) and benzo(a)pyrene $(B({\alpha})P)$ using Salmonella typhimurium reversion assay. On direct-acting mutagen (MNNG, 4NQO) and indirect-acting mutagen (Trp-P-1, $(B({\alpha})P)$, we observed higher inhibitory effect. Stepwise fractionation of the ethanol extract was done by using ether, chloroform, ethyl acetate, butanol and water to obtain effective fraction. Among them, water fractions $(100\;{\mu}g/plate)$ of Pinus thunbergii, Pinus rigida, Pinus densiflora and Pinus koraiensis showed high inhibition of 91.65%, 94.7%, 84.22% and 79.02%, respectively, on the mutagenicity of MNNG in Salmonella typhimurium TA100.

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Antioxidative and Antimutagenic Effects of Agaricus bisporus Ethanol Extracts (양송이버섯 추출물의 산화적 스트레스 억제효과 및 항돌연변이 효과)

  • Yun, Mi-Ja;Oh, Se-In;Lee, Mee-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.1
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    • pp.19-24
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    • 2009
  • This study was performed to investigate the antioxidative effects such as the inhibition of malondialdehyde (MDA) and bovine serum albumin (BSA) conjugation reaction, inhibition of $Fe^{2+}$-induced lipid peroxidation and the scavenging effect on 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical, as well as antimutagenic capacities as Ames test in ethanol extracts of Agaricus bisporus. Agaricus bisporus ethanol extracts inhibited $Fe^{2+}$-induced lipid peroxidation and scavenged DPPH radical. The $IC_{50}$ of Agaricus bisporus ethanol extracts were 78.63 mg/assay for inhibition of MDA with BSA conjugation reaction, 4.06 mg/ assay for inhibition of $Fe^{2+}$-induced lipid peroxidation and 1.08 mg/assay for scavenging effect on DPPH radical. So, among the methods used in this study, the most effective antioxidative capacity in ethanol extracts of Agaricus bisporus was the scavenging effect on DPPH radical. The indirect and direct antimutagenic effects of ethanol extracts of Agaricus bisporus were examined by Ames test using Salmonella Typhimurium TA98 and TA100. The inhibitory effects on direct mutagenicity mediated by sodium azide in Salmonella Typhimurium TA100 and 2-nitrofluorene in Salmonella Typhimurium TA98 were 100%. The inhibition rates on indirect mutagenicity mediated by 2-anthramine were 86.09% in the Salmonella Typhimurium TA98 and 81.93% in the Salmonella Typhimurium TA100. The ethanol extracts of Agaricus bisporus showed considerable antioxidative activity and strong antimutagenic capacity.

Rapid Detection for Salmonella spp. by Ultrafast Real-time PCR Assay (Ultrafast Real-time PCR법을 이용한 살모넬라의 신속 검출)

  • Kim, Seok Hwan;Lee, Yu-Si;Joo, In-Sun;Kwak, Hyo Sun;Chung, Gyung Tae;Kim, Soon Han
    • Journal of Food Hygiene and Safety
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    • v.33 no.1
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    • pp.50-57
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    • 2018
  • Salmonella continue to be a major cause of food poisoning worldwide. The rapid detection method of food-borne Salmonella is an important food safety tool. A real-time polymerase chain reaction (PCR) has been used as a rapid method for the detection of pathogens. It has been recently reported that NBS LabChip real-time PCR is a novel, ultrafast, and chip-type-convenient real-time PCR system. We developed the assay method based on NBS LabChip real-time PCR for the rapid detection of Salmonella, which its reaction time was within 20 minutes. Two target genes (invA and stn) were selected to design target specific primers and probes. The new method was validated by checking specificity and sensitivity (limit of detection). This study included forty-two target and twenty-one non-target strains to assess the specificity. This assay was able to identify the 42 Salmonella strains correctly. The limit of detection (LOD) was $10^1copies/{\mu}L$ in Salmonella genomes DNA, while LOD incubated for 4 hr in the inoculated sausage sample ranged from $10^1CFU/g$ to $10^2CFU/g$ as an inoculated cell count. The assay developed in this study could be applied for the investigation of food poisoning pathogens.