• Clinical and Experimental Vaccine Research
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• v.12 no.1
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• pp.70-76
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• 2023

Purpose: Typhoid remains a major health problem, especially in the developing world. Furthermore, the emergence of multidrug-resistant and extensively drug-resistant strains of Salmonella typhi added a sense of urgency to develop more effective typhoid vaccines, one of which is bacterial ghosts (BGs), prepared by both genetic and chemical means. The chemical method includes incubation with numerous agents for a short time at their minimum inhibitory or minimum growth concentrations. This study included the preparation of BGs by a sponge-like reduced protocol (SLRP). Materials and Methods: Critical concentrations of sodium dodecyl sulfate, NaOH, and H₂O₂ were used. Moreover, high-quality BGs were visualized by scanning electron microscope (SEM). Subculturing was used to confirm the absence of vital cells. Besides, the concentrations of the released DNA and protein were estimated spectrophotometrically. In addition, the integrity of cells was proved by visualizing Gram-stained cells using a light microscope. Furthermore, a comparison between the immunogenicity and safety of the prepared vaccine and the available whole-cell killed vaccine was established. Results: Improved preparation of high-quality BGs of S. typhi, visualized by SEM, revealed punctured cells with intact outer shells. Moreover, the absence of vital cells was confirmed by subculturing. At the same time, the release of respective amounts of proteins and DNA is another evidence of BGs' production. Additionally, the challenge test provided evidence that the prepared BGs are immunogenic and have the same efficacy as the whole cell vaccine. Conclusion: The SLRP provided a simple, economical, and feasible method for BGs preparation.

• Journal of Food Hygiene and Safety
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• v.18 no.4
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• pp.183-188
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• 2003

Bactericidal effect of $Green-Zone^{(TM)}$ was observed, when Staphylococcus aureus, E. coli O157:H7, Salmonella typhimurium, S. enteritidis, Listeria monocytogenes, the causative bacteria of food poisoning, Vibrio parahaemolyticus, and Shigella sonnei were treated with the diluted solution of $Green-Zone^{(TM)}$(33.3%~4.1%) for 30min at $20^{\circ}C$. All the bacteria were killed in 30 sec, when 33.3%-diluted $Green-Zone^{(TM)}$ was applied, except for S. aureus. Coronavirus, the same virus with SARS virus taxonomically, was also lilled with the 20%-diluted $Green-Zone^{(TM)}$. Canine parvovirus and Canine distermper virus were also killed even in the organic matter and hard water when treated with $Green-Zone^{(TM)}$. When applied to food such as raw fish, chilled meat, vegetables, $Green-Zone^{(TM)}$ could also decrease the number of microorganism, expecially for E. Coli. From these results, $Green-Zone^{(TM)}$ is thought to be effective for killing virus and bacteria, and also was proved to be safe when applied directrly to food.

• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.201-207
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• 2016

The aim of this study was to investigate the potential of lactic acid bacteria (LAB) strains as probiotics. Two strains were isolated from healthy chicken cecum and their acid and bile tolerance, residual organic acids, antibacterial activity against pathogenic bacteria, and immunomodulation activity were measured. Identification of the isolated strains was performed using the API 50CHL system and phylogenetic analysis using 16S rDNA sequencing. The isolates were determined to be Lactobacillus sakei strains. The acid tolerance of strains L2 and L8 was high enough that 75% of the inoculum survived in pH 2 for 2 h. The bile tolerance of both strains was observed at a 1% Oxgall concentration in MRS broth. The production of organic acids (lactic acid and acetic acid) and pH changes during growth were monitored and the maximum concentrations were obtained after 48 h of incubation. Culture supernatants of the two LAB strains showed strong antibacterial activity against pathogenic bacteria. The heat-killed LAB cells also induced high levels of immune cell proliferation compared with the control, and stimulated IL-6 and TNF-α production in mouse macrophages. Therefore, L. sakei strains L2 and L8 can be considered suitable probiotic bacteria.

• Journal of Microbiology and Biotechnology
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• v.28 no.6
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• pp.883-892
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• 2018

Probiotics, including Enterococcus faecium, confer a health benefit on the host. An Enterococcus strain was isolated from healthy chicken cecum, identified as E. faecium by 16S rDNA gene sequence analysis, and designated as E. faecium L11. To evaluate the potential of E. faecium L11 as a probiotic, the gastrointestinal tolerance, immunomodulatory activity, and lifespan extension properties of the strain were assayed. E. faecium L11 showed >66% and >62% survival in artificial gastric juice (0.3% pepsin, pH 2.5) and simulated small intestinal juice (0.5% bile salt and 0.1% pancreatin), respectively. Heat-killed E. faecium L11 significantly (p < 0.05) increased immune cell proliferation compared with controls, and stimulated the production of cytokines (IL-6 and $TNF-{\alpha}$) by activated macrophages obtained from ICR mice. In addition, E. faecium L11 showed a protective effect against Salmonella Typhimurium infection in Caenorhabditis elegans. In addition, feeding E. faecium L11 significantly (p < 0.05) extended the lifespan of C. elegans compared with the control. Furthermore, genes related to aging and host defense were upregulated in E. faecium L11-fed worms. In conclusion, E. faecium L11, which prolongs the lifespan of C. elegans, may be a potent probiotic supplement for livestock.

• Journal of Microbiology and Biotechnology
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• v.32 no.6
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• pp.816-823
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• 2022

The rapid spread of superbugs leads to the escalation of infectious diseases, which threatens public health. Endolysins derived from bacteriophages are spotlighted as promising alternative antibiotics against multi-drug resistant bacteria. In this study, we isolated and characterized the novel Salmonella typhimurium phage PBST08. Bioinformatics analysis of the PBST08 genome revealed putative endolysin ST01 with a lysozyme-like domain. Since the lytic activity of the purified ST01 was minor, probably owing to the outer membrane, which blocks accessibility to peptidoglycan, antimicrobial peptide cecropin A (CecA) was fused to the N-terminus of ST01 to disrupt the outer membrane. The resulting CecA::ST01 has been shown to have increased bactericidal activity against gram-negative pathogens including Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, and Enterobacter cloacae and the most affected target was A. baumannii. In the presence of 0.25 µM CecA::ST01, A. baumannii ATCC 17978 strain was completely killed and CCARM 12026 strain was wiped out by 0.5 µM CecA::ST01, which is a clinical isolate of A. baumannii and resistant to multiple drugs including carbapenem. Moreover, the larvae of Galleria mellonella could be rescued up to 58% or 49% by the administration of CecA::ST01 upon infection by A. baumannii 17978 or CCARM 12026 strain. Finally, the antibacterial activity of CecA::ST01 was verified using 31 strains of five gram-negative pathogens by evaluation of minimal inhibitory concentration. Thus, the results indicate that a fusion of antimicrobial peptide to endolysin can enhance antibacterial activity and the spectrum of endolysin where multi-drug resistant gram-negative pathogens can be efficiently controlled.

• The Journal of the Korean Society for Microbiology
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• v.21 no.4
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• pp.447-453
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• 1986

To diagnose the typhoid fever rapidly and accurately in clinically suspected patients, the levels of IgG subclass antibody were measured by enzyme-linked immunosorbent assay(ELISA). With symptom, blood culture and agglutination test, tested persons were categorized into 6 groups as typhoid fever, FUO, paratyphi A or B, other bacterial infctions, cancers, and control. ELISA was performed on the polyvinyl chloride plates coated with killed whole cell($10^8\;cell/ml$) of S. typhi 0901W by poly-L-lysine applied as binding substance (and polyvinyl chloride as solid phase). The distribution of the level of IgG subclass antibodies in each group was analyzed and compared with other groups. The results obtained were summarized as follow: 1. The optimal dilution of the sera from patients with typhoid fever was 1:160, and those of the sheep anti-human IgG subclass and the peroxidase conjugated rabbit anti-sheep IgG were 1:4000 and 1:5000, respectively. 2. The absorbance levels of IgG subclass in the sera of typhoid fever patients were as follows; a) IgG1 value is $0.439{\pm}0.110$ b) IgG2 value is $0.416{\pm}0.165$ c) IgG3 value is $0.449{\pm}0.145$ d) IgG4 value is $0.525{\pm}0.154$ IgG subclass levels in the sera of typhoid patients were much higher than in control group and patient with paratyphi A or B as well as other infectious diseases. The sensitivity and the specificity in differential diagnosis of typhoid fever and other febrile diseases were 92% and 79% in the assay of IgG1 respectively, whereas those in the assay of IgG2 were 97% and 72%, respectively (above absorbance 0.3). 3. The absorbance levels of IgG subclass in the serial sera of typhiod fever patients tend to decrease to the level of absorbance 0.3 in 10 months from the onset of illness. 4. The order of absorbance levels of IgG subclass in the serum of each group were typhoid fever, paratyphi A or B, other infectious diseases, control and cancer. 5. For the serodiagnosis of typhoid fever against other febrile diseases, the sensitivity and the specificity in the assay of IgG2 activity were 76% and 93% in absorbance 0.4, respectively. 6. In the distribution of the level of each IgG subclass in the sera of FUO patients which were suspected of typhoid fever, the positive rate was ranged from 36% to 82%. This suggest that more than 50% of FUO patients are caused by S. typhi.

• Korean Journal of Food Science and Technology
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• v.14 no.4
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• pp.291-300
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• 1982

In order to develop effective and simple sanitation method for the extention of shelf-life of fresh poultry meat, the effect of sanitizers, sanitation methods and packaging materials on the extention of shelf-life of poultry meats was observed at the $4^{\circ}C$ and room temp$(10{\sim}20^{\circ}C)$. The results are summarized as follows: 1. The autochonous skin microflora of poultry, before processing, were believed to be removed or killed during the scalding and plucking, and exposed dermal tissue was contaminated by microorganisms from the subsequent stages of processing. 2. In the final stage of poultry processing, total viable counts of microorganisms and coliforms were averaged to $3.5{\times}10^4/cm^2$ and $400/cm^2$, respectively. 3. The refrigerated shelf-life of fresh whole poultry carcasses at $3\;to\;4^{\circ}C$ was extended to 7 to 16 days compared to control with the various treatments of some sanitizers by dipping freshly chilled carcasses for 5 min or spraying 1 liter of sanitizers per carcasses. In the case of storage at $10\;to\;15^{\circ}C$, the shelf-life of poultry carcasses was extended to one to two days by the sanitation treatments compared to control. 4. Spraying sanitation was more effective than dipping sanitation, and 5 minutes dipping and one liter spraying per carcass were enough for effective sanitation of poultry carcasses in most sanitizers. 5. The packaging with an oxygen impermeable polyvinylidene chloride extended the shelf-life to 10 days and 5 days with polyethylene compared to control. When poultry carcasses were sanitized by continuous spraying with one liter of 30 ppm of chlorine and another one liter of 5% of potassium sorbate, packaged with polyvinylidene chlorlde were extended to about 30 days compared to control.