• Korean Journal of Veterinary Service
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• v.32 no.2
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• pp.147-153
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• 2009

Salmonella species are the most important etiologic agents of food-borne acute gastroenteritis. The most common serotypes isolated from humans are Salmonella enterica serotype Typhimurium (S. Typhimurium) and S. Enteritidis. Traditional detection methods for Salmonella are based on cultures using selective media and characterization of suspicious colonies by biochemical and serological tests. These methods are generally time-consuming and not so highly sensitive. Recently, the polymerase chain reaction (PCR) has been used as a highly sensitive, specific, and rapid test for the presence of pathogenic bacteria. In this study, a multiplex PCR (m-PCR) was used to detect S. Typhimurium and S. Enteritidis. We selected m-PCR target genes, which were the spv (virulence plasmid specific for S. Enteritidis) and sefA (S. Enteritidis fimbrial antigen) genes, fliC (H1-i antigen specific for S. Typhimurium) and a randomly cloned sequence specific for the genus Salmonella. With m-PCR, random sequence was detected from all strains of Salmonella spp, spv and sefA were detected from all strains of S. Enteritidis (100%), and fliC was detected from all strains of S. Typhimurium (100%). This assay indicate that the specificity of the m-PCR make them potentially valuable tools for detection of S. Typhimurium and S. Enteritidis.

• Journal of Environmental Health Sciences
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• v.39 no.3
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• pp.239-246
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• 2013

Objectives: Antimicrobial resistance and multidrug resistance patterns have been studied with a total of 189 samples of Salmonella Enteritidis and Salmonella Typhimurium isolated from diarrhea patients in Incheon from 2008 to 2012. Methods: Antimicrobial resistance tests were determined by Disc Diffusion method. Results: The serological distribution of Salmonella spp. showed 108 strains (30.1%) of S. Enteritidis, 81 strains (22.6%) of S. Typhimirium, eight strains (8.0%) of S. Typhi, 11 strains ( 3.1% ) of S. Paratyphi, and the 151 other strains (42.1%). The separation rate of Salmonella spp. by year showed 14.5% (52 strains) in 2008, 13.6% (49 strains) in 2009, 22.8% (82 strains) in 2010, 25.3% (91 strains) in 2011, and 23.7% (85 strains) in 2012. Additionally, the separation rate of S. Enteritidis and S. Typhimirium in 2010 was the highest. The Salmonella spp. isolated from diarrhea patients showed significant differences according to age (p<0.05), gender (p<0.01) and medical institution (p<0.05). The highest resistance was found to the following antimicrobial agents: imipenem 77 strains, ampicillin 47 strains, ciprofloxacin 34 strains, nalidixic acid 29 strains for S. Enteritidis, and ampicillin 45 strains, nalidixic acid 45 strains for S. Typhimurium. Separated S. Enteritidis and S. Typhimurium resistance to the antibiotics by the year showed significant differences (p<0.05). The patterns of multidrug resistance rates were 43.1% (47 strains) for one drug, 8.3% (9 strains) for two drugs, 11.0% (12 strains) for three drugs, 15.62% (17 strains) for four drugs, and 13.7% (15 strains) for five or more drugs for S. Enteritidis. For S. Tyhpimurium, the rates were 15.0% (12 strains) for one drug, 10.0% (8 strains) for two drugs, 6.3% (five strains) for three drugs, 18.7% (15 strains) for four drugs, and 23.8% (19 strains) for five or more drugs. Conclusion: The antibiotic resistance issue is directly related to people's lives. Thus, the usage of antibiotics should be reduced in order to manage antibiotic resistance.

• Journal of Microbiology and Biotechnology
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• v.27 no.8
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• pp.1519-1528
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• 2017

Foodborne contamination and salmonellosis caused by Salmonella Enteritidis (S. Enteritidis) are a significant threat to human health and poultry enterprises. Outer membrane vesicles (OMVs), which are naturally secreted by gram-negative bacteria, could be a good vaccine option because they have many biologically active substances, including lipopolysaccharides (LPS), outer membrane proteins (OMPs), and phospholipids, as well as periplasmic components. In the present study, we purified OMVs derived from S. Enteritidis and analyzed their characteristics through silver staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis. In total, 108 proteins were identified in S. Enteritidis OMVs through liquid chromatography tandem mass spectrometry analysis, and OMPs, periplasmic proteins, and extracellular proteins (49.9% of total proteins) were found to be enriched in the OMVs compared with bacterial cells. Furthermore, native OMVs used in immunizations by either the intranasal route or the intraperitoneal route could elicit significant humoral and mucosal immune responses and provide strong protective efficiency against a lethal dose (~100-fold $LD_{50}$) of the wild-type S. Enteritidis infection. These results indicated that S. Enteritidis OMVs might be an ideal vaccine strategy for preventing S. Enteritidis diseases.

• Journal of Food Hygiene and Safety
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• v.15 no.3
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• pp.262-266
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• 2000

The polymerase chain reaction was used to selectively detect sequences within the fimbrial antigen of Salmonella enteritidis. Sterile milk was artificially inoculated with known amount of S. enteritidis and then DNA was extracted with guanidine thiocyanate/phenol/chloroform, followed by PCR. A detection limit of as few as 100 colony forming unit (cfu) per 0.5 ml milk was obtained with this method. For the whole procedure, it took only 5 h. A semi-quantitative polymerase chain reaction assay which allows an estimation of colony forming unit of S. enteritidis was developed. Known amount of standard plasmid pGem-4Z-Sef B(-) containing cloned S. enteritidis fimbrial antigen gene was co-amplified with Salmonella genomic DNA isolated from artificially inoculated milk. The same set of primers were used for the amplification and the products were cleaved with Bam HI. The concentration of the target DNA could be estimated by comparing the intensity of the two bands after electrophoresis. The PCR-based protocol described in this paper provides a rapid, simple, and sensitive method for detecting S. enteritidis in milk.

• Journal of Dairy Science and Biotechnology
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• v.33 no.4
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• pp.271-275
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• 2015

Colloidal semiconductor CdSe-ZnS core-shell nanocrystal quantum dot (Qdot) are luminescent inorganic fluorophores that show potential to overcome some of the functional limitations encountered with organic dyes in fluorescence labeling applications. Salmonella Enteritidis has emerged as a major cause of human salmonellosis worldwide since the 1980s. A rapid, specific, and sensitive method for the detection of Salmonella Enteritidis was developed using Qdot as a fluorescence marker coupled with immunomagnetic separation. Magnetic beads coated with anti-Salmonella Enteritidis antibodies were employed to selectively capture the target bacteria, and biotin-conjugated anti-Salmonella antibodies were added to form sandwich immune complexes. After magnetic separation, the immune complexes were labeled with Qdot via biotin-streptavidin conjugation, and fluorescence measurement was carried out using a fluorescence measurement system. The detection limit of the Qdot method was a Salmonella Enteritidis concentration of $10^3$ colony-forming units (CFU)/mL, whereas the conventional fluorescein isothiocyanate-based method required over $10^5CFU/mL$. The total detection time was within 2 h. In addition to the potential for general nanotechnology development, these results suggest a new rapid detection method of various pathogenic bacteria from a complex food matrix.

• Journal of Biosystems Engineering
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• v.35 no.2
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• pp.116-123
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• 2010

Rapid detection of foodborne pathogens has been a major challenge for the food industry. Salmonella contamination is well known in all foods including pasteurised milk. The possibility of specific detection of Salmonella Enteritidis by surface plasmon resonance (SPR) biosensor was explored using a commercially available portable SPR sensor. Self assembly technique was adopted to immobilize anti-Salmonella antibodies on the gold sensing surface of the SPR sensor. The concentration of polyclonal antibody for use in the SPR biosensor was chosen to 1.0 mg/mL. Experiments were conducted at near real-time with results obtained for one SPR biosensor assay within 1 hour. The limit of detection for Salmonella Enteritidis was determined to be $10^6$ CFU/mL in both PBS buffer and milk samples. The assay sensitivity was not significantly affected by milk matrix. Our results showed that it would be possible for employing the SPR biosensor to detect Salmonella Enteritidis in near real-time.

• Korean Journal of Veterinary Research
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• v.40 no.3
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• pp.505-514
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• 2000

This study was conducted to investigate the isolation prequency, serotypes, and related epidemiological properties of 341 Salmonella spp from domestic poultry and environmental samples during the period 1993-1995. A total of 1,918 samples was collected during the three years period in nationwide. Most of Salmonella spp were isolated from the intestinal contents of poultry, especially cecal(46.0%) and rectal(35.8%) contents. Among the tested samples, rat(28.5 %) was the most predominant Salmonella reservoirs and followed by duck(24.8%), broiler(18.8%), layer(14.8%) and feed(7.1%), in order. More than twelve Salmonella serovars were identified among the 341 Salmonella isolates. The most prevalent serotypes isolated from non-human sources were S enteritidis (22.3%) and S pullorum (21.9%), S muenchen (13.9%), S typhimurium (12.6%), S gallinarum, S meleagridis, S heidelberg, and S senftenberg were followed, in order. In layer chickens, S pullorum (26.0%) was the most predominant serotype but S muenchen (33.0%) was in broiler chickens, S enteritidis (28.4%) was in ducks, and S typhimurium (60.0%) was in rats, respectively. As a results, S enteritidis was identified as the most prevalent serotype in non-human Salmonella isolates in Korea during the period 1993-1995. A preliminary study on the phage typing of 19 S enteritidis selected from the nationwide scale was shown that S enteritidis phage type(PT) 4 was the most predominant PT, and SEPT 1, SEPT 6a, SEPT 7 and SEPT 7a variant were also found in the same period.

• Journal of Food Hygiene and Safety
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• v.31 no.5
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• pp.342-348
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• 2016

The objective of this study was to compare the change of S. Enteritidis with S. Typhimurium populations in liquid egg products. S. Enteritidis or S. Typhimurium was inoculated into egg white and egg yolk and stored at 8, 10, 15, 25, and $35^{\circ}C$, respectively. In egg white, no growth of S. Enteritidis and S. Typhimurium was observed at 8, 10, 15, and $35^{\circ}C$, while both S. Enteritidis and S. Typhimurium in egg white stored grew more than 1 log CFU/ml after 50 hours storage at $25^{\circ}C$. In egg yolk, there was no growth of S. Enteritidis and S. Typhimurium at $8^{\circ}C$ but growth of both strains was observed at 10, 15, 25, and $35^{\circ}C$. Since growth of S. Enteritidis and S. Typhimurium was only observed in egg yolk, primary growth models for both strains were developed using modified Gompertz equation and then secondary models for lag time (LT), specific growth rate (SGR), and maximum population density (MPD) were developed as a function of temperature. At all temperatures, more rapid growth of S. Enteritidis than S. Typhimurium was observed in egg yolk, indicating the greater risk of S. Enteritidis than S. Typhimurium in egg products. In conclusion, the results indicate that temperature control less than $8^{\circ}C$ is very important to ensure safety of liquid egg products, especially liquid egg yolk.

• Korean Journal of Veterinary Service
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• v.34 no.3
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• pp.217-226
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• 2011

Salmonella spp. is of increasing public health concern as causative pathogens of food poisoning. The aim of this study was to investigate the serotypes and antimicrobial resistance pattern of Salmonella spp. isolated from duck farms in Daegu-Gyeongbuk province. Also, S. Enteritidis and S. Typhimurium isolates were further examined for plasmid analysis, phage typing and pulsed-field gel electrophoresis (PFGE). A total of 34 Salmonella spp. (16.4%) were isolated from duck farms and ten serotypes were identified. The predominant serotypes were S. Typhimurium (23.5%) S. Fyris (17.6%) and S. Haardt (11.8%), S. Agona and S. Enteritidis (respectively 8.8%). Of 34 Salmonella isolates, 15 (44.1%) isolates were resistant to at least one antimicrobial agent and multiple resistance (resistance to more than 4 drugs) was observed in 9 strains (26.5%). The high resistance was found to streptomycin (32.4%), tetracycline (29.4%), ampicillin, kanamycin and nalidixic acid (respectively, 26.5%), all Salmonella isolates were susceptible to cefoxitin, cefotaxime, gentamicin, amikacin and ciprofloxacin. All S. Enteritidis and S. Typhimurium isolates were found to contain only one plasmid (ca. 54 or 55kb, respectively). Among the S. Enteritidis isolates, two phage types were found, PT32a and PT1c, respectively, one isolates did not react with any of the phages used. Whereas, all S. Typhimurium isolates were RDNC (reacts but does not conform). PFGE showed to be a useful typing method better than plasmid analysis and phage typing for discrimination of isolates especially, S. Typhimurium isolates. Our results indicated that the serotypes of Salmonella isolates are widely distributed in duck farms, further epidemiological studies should be carried out.

• 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.