• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.454-464
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• 2019

Salmonellosis is a highly contagious bacterial disease that threatens both human and poultry health. Tests that can detect Salmonella in the field are urgently required to facilitate disease control and for epidemiological investigations. Here, we combined loop-mediated isothermal amplification (LAMP) with a chromatographic lateral flow dipstick (LFD) to rapidly and accurately detect Salmonella. LAMP primers were designed to target the Salmonella invA gene. LAMP conditions were optimized by adjusting the ratio of inner to outer primers, $MgSO_4$ concentration, dNTP mix concentration, amplification temperature, and amplification time. We evaluated the specificity of our novel LAMP-LFD method using six Salmonella species and six related non-Salmonella strains. All six of the Salmonella strains, but none of the non-Salmonella strains, were amplified. LAMP-LFD was sensitive enough to detect concentrations of Salmonella enterica subsp. enterica serovar Pullorum genomic DNA as low as $89fg/{\mu}l$, which is 1,000 times more sensitive than conventional PCR. When artificially contaminated feed samples were analyzed, LAMP-LFD was also more sensitive than PCR. Finally, LAMP-LFD gave no false positives across 350 chicken anal swabs. Therefore, our novel LAMP-LFD assay was highly sensitive, specific, convenient, and fast, making it a valuable tool for the early diagnosis and monitoring of Salmonella infection in chickens.

• Journal of Veterinary Science
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• v.24 no.6
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• pp.82.1-82.12
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• 2023

Background: The current conventional serotyping based on antigen-antisera agglutination could not provide a better understanding of the potential pathogenicity of Salmonella enterica subsp. enterica serovar Brancaster. Surveillance data from Malaysian poultry farms indicated an increase in its presence over the years. Objective: This study aims to investigate the virulence determinants and antimicrobial resistance in S. Brancaster isolated from chickens in Malaysia. Methods: One hundred strains of archived S. Brancaster isolated from chicken cloacal swabs and raw chicken meat from 2017 to 2022 were studied. Two sets of multiplex polymerase chain reaction (PCR) were conducted to identify eight virulence genes associated with pathogenicity in Salmonella (invasion protein gene [invA], Salmonella invasion protein gene [sipB], Salmonella-induced filament gene [sifA], cytolethal-distending toxin B gene [cdtB], Salmonella iron transporter gene [sitC], Salmonella pathogenicity islands gene [spiA], Salmonella plasmid virulence gene [spvB], and inositol phosphate phosphatase gene [sopB]). Antimicrobial susceptibility assessment was conducted by disc diffusion method on nine selected antibiotics for the S. Brancaster isolates. S. Brancaster, with the phenotypic ACSSuT-resistance pattern (ampicillin, chloramphenicol, streptomycin, sulphonamides, and tetracycline), was subjected to PCR to detect the corresponding resistance gene(s). Results: Virulence genes detected in S. Brancaster in this study were invA, sitC, spiA, sipB, sopB, sifA, cdtB, and spvB. A total of 36 antibiogram patterns of S. Brancaster with a high level of multidrug resistance were observed, with ampicillin exhibiting the highest resistance. Over a third of the isolates displayed ACSSuT-resistance, and seven resistance genes (β-lactamase temoneira [bla_TEM], florfenicol/chloramphenicol resistance gene [floR], streptomycin resistance gene [strA], aminoglycoside nucleotidyltransferase gene [ant(3")-Ia], sulfonamides resistance gene [sul-1, sul-2], and tetracycline resistance gene [tetA]) were detected. Conclusion: Multidrug-resistant S. Brancaster from chickens harbored an array of virulence-associated genes similar to other clinically significant and invasive non-typhoidal Salmonella serovars, placing it as another significant foodborne zoonosis.

• Korean Journal of Microbiology
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• v.54 no.2
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• pp.164-166
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• 2018

Salmonella enterica subsp. enterica is a foodborne pathogen that has been detected throughout the world. Here, we present the complete genome sequence of Salmonella Enteritidis isolated from a commercial kimbap that caused foodborne illness in the Republic of Korea in 2014. Complete genome sequence analysis of Salmonella Enteritidis MFDS1004839 revealed a 4,679,649 bp chromosome and a 96,994 bp plasmid, with G + C contents of 52.2% and 49.3%, respectively. The chromosome and plasmid genome included 4,482 predicted protein-coding sequences, 84 tRNAs and 22 rRNAs genes.

• Journal of Life Science
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• v.11 no.5
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• pp.432-438
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• 2001

Various lactic acid bacteria were isolated from Korean Dongchimi (whole radish Kimichi with added water) in order to study their antimutagenic activity. Ames test using Salmonella enterica serovar typhimurium TA98 and TA100 showed the strain DLAB19 to have the highest antimutagenic activity among the 300 isolated strains against MNNG(N-methyl-N-nitro-N-nitrosoguanidine), NPD (4-nitro-O-phenylenediamine), 4-NQO(4-nitroquinoline-1-oxide) and AFB$_{1}$(aflatoxin B$_{1}$). The strain was identified as Leuconostoc mesenteroides subsp. cremoris according to the Bergeys Mannual Systematic Bscteriology based on its morphological, cultural, physiological characteristics and biological system Antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was found in the culture supernatant suggesting the bacterium secretes, the antimutagenic substance in the media. The antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was reconfirmed by the spore-rec assay using spores of Bacillus subtilis H17 (Rec$^{+}$) and M45 (Rec$^{[-10]}$ ).).

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.7
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• pp.1009-1016
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• 2014

This study investigated the probiotic and functional characteristics of Mukeunji starter, Lactobacillus curvatus ML17, isolated from Mukeunji. Lb. curvatus ML17 was confirmed as a safe microorganism due to its non-hemolytic activity and non-production of harmful ${\beta}$-glucuronidase and ${\beta}$-glucosidase. Tolerance to artificial gastric and bile juice of Lb. curvatus ML17 was investigated. After incubation in artificial gastric and bile juice, the number of surviving cells was $1.38{\times}10^8$ CFU/mL. According to the results of adhesion assay, this strain also exhibited good adhesion to Caco-2 cells. Lb. curvatus ML17 showed good antimicrobial activity against food borne pathogens, especially Micrococcus luteus, Bacillus cereus, Salmonella enterica subsp. enterica, and Pseudomonas aeruginosa. Cell-free extract of Lb. curvatus ML17 exhibited high levels of DPPH scavenging capacity and inhibitory effects on growth of AGS human gastric adenocarcinoma cells and HT-29 human colon carcinoma cells. These results suggest that Lb. curvatus ML17 has potential for application in functional foods.

• Environmental Analysis Health and Toxicology
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• v.30
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• pp.14.1-14.7
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• 2015

Objectives Cigarette smoking is associated with carcinogenesis owing to the mutagenic and genotoxic effects of cigarette smoke. The aim of this study was to evaluate the mutagenic and genotoxic effects of Korean cigarettes using in vitro assays. Methods We selected 2 types of cigarettes (TL and TW) as benchmark Korean cigarettes for this study, because they represent the greatest level of nicotine and tar contents among Korean cigarettes. Mutagenic potency was expressed as the number of revertants per ${\mu}g$ of cigarette smoke condensate (CSC) total particulate matter whereas genotoxic potency was expressed as a concentration-dependent induction factor. The CSC was prepared by the International Organization for Standardization 3308 smoking method. CHO-K1 cells were used in vitro micronucleus (MNvit) and comet assays. Two strains of Salmonella typhimurium (Salmonella enterica subsp. enterica ; TA98 and TA1537) were employed in Ames tests. Results All CSCs showed mutagenicity in the TA98 and TA1537 strains. In addition, DNA damage and micronuclei formation were observed in the comet and MNvit assays owing to CSC exposure. The CSC from the 3R4F Kentucky reference (3R4F) cigarette produced the most severe mutagenic and genotoxic potencies, followed by the CSC from the TL cigarette, whereas the CSC from the TW cigarette produced the least severe mutagenic and genotoxic potencies. Conclusions The results of this study suggest that the mutagenic and genotoxic potencies of the TL and TW cigarettes were weaker than those of the 3R4F cigarette. Further study on standardized concepts of toxic equivalents for cigarettes needs to be conducted for more extensive use of in vitro tests.

• Korean Journal of Veterinary Service
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• v.27 no.1
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• pp.7-15
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• 2004

For measuring general bacterial count and Escherichia coli count, the standard plate count was used to conduct a test on 113 cases of pork carcass surfaces from slaughterhouses in Incheon area from January to February 2003. And Salmonella spp, Staphylococcus aureus and Listeria monocytogenes was used to conduct a test on 68 cases of pork carcass surfaces and 76 cases of feces, that is to say, a total of 144 cases. The results were obtained as follows: In the case of general bacterial count, 29 cases(25.7%) were in the range of 100∼999 and 62 cases(54.9%) were in the range of 1,000∼9,999 and 22 cases(19.4%) were in the range of 10,000∼99,999. Meanwhile as regards E coli count, 22 cases(19.4%) were in the range of 1∼9 and 69 cases(61.2%) were in the range of 10∼99 and 22 cases(19.4%) were in the range of 100∼999. On 68 cases of pork carcass surfaces, 7 strains(10.2%) of Salmonella spp, and 12 strains(17.6%) of S aureus were detected and 27 strains(39.7%) of L monocytogenes, respectively. As for the detected Salmonella spp, 6 strains of the B group, 3 strains of S enterica subsp salame and 2 strains of S typhimurium were detected, respectively. On 76 cases of feces, 14 strains(18.4%) of Salmonella spp, and 15 strains(19.7%) of L monocytogenes and 14 strains(18.4%) of S aureus were detected respectively. As for the detected Salmonella spp, 6 strains of the B group, 4 strains of S derby and 8 strains of the C group, 5 strains of S rissen were detected, respectively. All of 42 strains of L monocytogenes were type 1. As a result of conducting a toxin test on the detected S aureus, all of 26 strains were found to be non-toxin.

• Food Engineering Progress
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• v.14 no.4
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• pp.316-321
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• 2010

The efficacy of the ozonated water (0.1, 0.2, 0.4, 0.6, and 1.0 ppm) in reducing the risk of food-borne disease was investigated in this study. After inoculation of Listeria monocytogenes (ATCC 19112), Salmonella enterica subsp. enterica biovar Typhimurium (ATCC 12598), Escherichia coli O157:H7 (ATCC 43890) to lettuce, spinach, and beef, inhibition effect with different washing concentrations, time, and methods with ozonated and tap water were evaluated. As a result, there were 2.16 to 3.85 log CFU/mL reduction in different foods and 7 log CFU/mL reduction on cutting boards after watering with ozonated water. Higher than 0.2 ppm of ozonated water treatment reduced the growth of food-borne disease bacteria with increasing washing time and ozone concentration. These results suggested that the ozonated water treatment effectively improved the microbiological quality and food safety.

• Journal of Animal Science and Technology
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• v.62 no.6
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• pp.912-921
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• 2020

We examined the rates of pathogenic bacterial cross-contamination from gloves to meat and from meat to gloves during pork processing under meat-handling scenarios in transfer rate experiments of inoculated pathogens. The inoculated pork contained ~5-6 Log10 CFU/g pathogenic bacteria like Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), Listeria monocytogenes (L. monocytogenes), and Salmonella enterica subsp. enterica (Sal. enteritidis). On cotton gloves, after cutting the pork, the cutting board, knife, and cotton gloves showed 3.07-3.50, 3.29-3.92 and 4.48-4.86 Log10 CFU/g bacteria. However, when using polyethylene gloves, fewer bacteria (3.12-3.75, 3.20-3.33, and 3.07-3.97 Log10 CFU/g, respectively) were transferred. When four pathogens (6 Log10 CFU/g) were inoculated onto the gloves, polyethylene gloves showed a lower transition rate (cutting board 2.47-3.40, knife 2.01-3.98, and polyethylene glove 2.40-2.98 Log10 CFU/g) than cotton gloves. For cotton gloves, these values were 3.46-3.96, 3.37-4.06, and 3.55-4.00 Log10 CFU/g, respectively. Use of cotton gloves, polyethylene gloves, knives and cutting boards for up to 10 hours in a meat butchering environment has not exceeded HACCP regulations. However, after 10 h of use, 3.09, 3.27, and 2.94 Log10 CFU/g of plate count bacteria were detected on the cotton gloves, cutting board, and knives but polyethylene gloves showed no bacterial count. Our results reveal the transfer efficiency of pathogenic bacteria and that gloved hands may act as a transfer route of pathogenic bacteria between meat and hands. The best hand hygiene was achieved when wearing polyethylene gloves. Thus, use of polyethylene rather than cotton gloves reduces cross-contamination during meat processing.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.5
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• pp.620-631
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• 2015

A total of 59 lactic acid bacteria (LAB) strains were isolated from corn stover silage. According to phenotypic and chemotaxonomic characteristics, 16S ribosomal DNA (rDNA) sequences and recA gene polymerase chain reaction amplification, these LAB isolates were identified as five species: Lactobacillus (L.) plantarum subsp. plantarum, Pediococcus pentosaceus, Enterococcus mundtii, Weissella cibaria and Leuconostoc pseudomesenteroides, respectively. Those strains were also screened for antimicrobial activity using a dual-culture agar plate assay. Based on excluding the effects of organic acids and hydrogen peroxide, two L. plantarum subsp. plantarum strains ZZU 203 and 204, which strongly inhibited Salmonella enterica ATCC $43971^T$, Micrococcus luteus ATCC $4698^T$ and Escherichia coli ATCC $11775^T$ were selected for further research on sensitivity of the antimicrobial substance to heat, pH and protease. Cell-free culture supernatants of the two strains exhibited strong heat stability (60 min at $100^{\circ}C$), but the antimicrobial activity was eliminated after treatment at $121^{\circ}C$ for 15 min. The antimicrobial substance remained active under acidic condition (pH 2.0 to 6.0), but became inactive under neutral and alkaline condition (pH 7.0 to 9.0). In addition, the antimicrobial activities of these two strains decreased remarkably after digestion by protease K. These results preliminarily suggest that the desirable antimicrobial activity of strains ZZU 203 and 204 is the result of the production of a bacteriocin-like substance, and these two strains with antimicrobial activity could be used as silage additives to inhibit proliferation of unwanted microorganism during ensiling and preserve nutrients of silage. The nature of the antimicrobial substances is being investigated in our laboratory.