• Clinical and Experimental Vaccine Research
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• v.12 no.4
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• pp.304-312
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• 2023

Purpose: Due to the many problems with commercially available vaccines, the production of effective vaccines against brucellosis is a necessity. The aim of this study was to evaluate the immune responses caused by the chimeric protein consisting of trigger factor, Bp26, and Omp31 (TBO) along with aluminum hydroxide (AH/TBO) and selenium (Se/TBO) nanoparticles (NPs) as adjuvants in mouse model. Materials and Methods: Recombinant antigen expression was induced in Escherichia coli BL21 (DE3) bacteria using IPTG (isopropyl-d-1-thiogalactopyranoside). Purification and characterization of recombinant protein was conducted through NiFe₃O₄ NPs, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Western blot. NP characteristics, including morphology and particle size, were measured in vitro. The recombinant TBO was loaded on to AH and Se NPs and were administered subcutaneously. After mice immunization, measurement of antibody titter and protection assay was performed. Results: The average sizes of AH and Se NPs were about 60 nm and 150 nm, respectively. The enzyme-linked immunosorbent assay results showed that the serum of mice immunized by subcutaneous injection with both nanovaccines produced significant immunoglobulin G (IgG) responses against the chimeric antigen. The results of TBO-specific IgG isotype (IgG2a/IgG1) analysis showed that both AH and Se NPs induced a type to T-helper immune response. In addition, the results of the challenge with the pathogenic strain of Brucella melitensis 16M showed that vaccinated mice with AH/TBO NPs indicated a higher reduction of bacterial culture than immunized mice with Se/TBO NPs and TBO alone. Conclusion: The results showed that AH NPs carrying chimeric antigen can be a promising vaccine candidate against brucellosis by producing protective immunity.

• Journal of Microbiology and Biotechnology
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• v.34 no.8
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• pp.1599-1608
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• 2024

Yersinia enterocolitica is a globally distributed food-borne gastrointestinal pathogen. The O-antigen variation-determined serotype is an important characteristic of Y. enterocolitica, allowing intraspecies classification for diagnosis and epidemiology purposes. Among the 11 serotypes associated with human yersiniosis, O:3, O:5,27, O:8, and O:9 are the most prevalent, and their O-antigen gene clusters have been well defined. In addition to the O-antigen, several virulence factors are involved in infection and pathogenesis of Y. enterocolitica strains, and these are closely related to their biotypes, reflecting pathogenic properties. In this study, we identified the O-AGC of a Y. enterocolitica strain WL-21 of serotype O:10, and confirmed its functionality in O-antigen synthesis. Furthermore, we analyzed in silico the putative O-AGCs of uncommon serotypes, and found that the O-AGCs of Y. enterocolitica were divided into two genetic patterns: (1) O-AGC within the hemH-gsk locus, possibly synthesizing the O-antigen via the Wzx/Wzy dependent pathway, and (2) O-AGC within the dcuC-galU-galF locus, very likely assembling the O-antigen via the ABC transporter dependent pathway. By screening the virulence genes against genomes from GenBank, we discovered that strains representing different serotypes were grouped according to different virulence gene profiles, indicating strong links between serotypes and virulence markers and implying an interaction between them and the synergistic effect in pathogenicity. Our study provides a framework for further research on the origin and evolution of O-AGCs from Y. enterocolitica, as well as on differences in virulent mechanisms among distinct serotypes.

• Journal of Food Hygiene and Safety
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• v.29 no.4
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• pp.268-277
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• 2014

This study assessed microbiological hazards at postharvest stage of dropwort farms (A, B, C, D, E, F, G, H, I) located in 4 different areas in Korea. The samples were assessed for sanitary indication bacteria (total aerobic bacteria, coliform, and Escherichia coli) and pathogenic bacteria (Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus). Total aerobic bacteria and coliform in 9 dropwort farms were detected at the levels of 0~7.00 and 0~4.25 log CFU/g, mL, of $100cm^2$. In particular, microbial contamination in worker's hand showed higher than cultivation environment factors. Escherichia coli was detected in several farms of soil, irrigation water, washing water and worker's hand and also, dropwort in these farms was contaminated with E. coli (positive reaction). In case of pathogenic bacteria, B. cereus was detected at the highest levels in soil. S. aureus was detected qualitatively from only one sample of dropwort washed by water. E. coli O157:H7 and L. monocytogenes were not detected. Although dropwort pass through 2 process (trimming and washing), the microbial contamination was not differ significantly before and after which indicates that current washing system was not effect on reduction of microorganism. From these results, the postharvest environment and workers have been considered as cross-contamination factors. Thus, processing equipments and personal hygiene should be managed to reduce the microbial contamination of dropwort. Accordingly management system such as good agricultural practices (GAP) criteria is needed for the safety of dropwort

• Journal of Life Science
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• v.33 no.10
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• pp.820-827
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• 2023

Since COVID-19 began at the end of 2019, the wearing time of protective clothing used to prevent pathogenic bacteria and virus infection has increased, and the development of safe protective materials that are human-friendly and have antibacterial and antiviral functions has been required. In this study, we investigated the possibility of developing natural antibacterial protection materials using ethanol extract of the medicinal plant Zanthoxylum Piperitum DC. The antibacterial activity assay of the 80% ethanol extract of Z. piperitum DC leaves against various nosocomial infectious bacteria, using the disk diffusion method, showed that Staphylococcus aureus ATCC 25923, Klebsiella pneumoniae ATCC 13883, Salmonella typhimurium, and Aeromonas hydrophila are sensitive to the inhibitory action of the extract. The IC₅₀ values of the ethanol extract against S. aureus, K. pneumoniae, P. vulgaris and A. hydrophila were about 0.59 mg/ml, 0.50 mg/ml, 1.06 mg/ml, and 0.06 mg/ml, respectively. To determine whether the ethanol extract of Z. piperitum DC leaves can be applied to the development of antibacterial protective fabric, the ethanol extract was tested using a protective fabric from the KM Health Care Corp. using the JIS L1902-Absorption method. As a result, the bacteriostatic and bactericidal activity values of S. aureus ATCC 25923 and K. pneumoniae ATCC 13883 appeared to be more than 2.0 when treated with the ethanol extract at a concentration of 1% (w/v). Together, these results suggest that Z. piperitum DC leaves can be applied to develop natural antibacterial functional protective fabrics.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.62 no.1
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• pp.1-8
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• 2017

We investigated the effect of silicate coating of rice seeds on bakanae disease incidence and the quality of seedlings raised in seedling boxes and transplanted into pots. The silicate-coated rice seed (SCS) was prepared as follows. Naturally infested rice seeds not previously subjected to any fungicidal treatment were dressed with a mixture of 25% silicic acid at pH 11 and 300-mesh zeolite powder at a ratio of 50 g dry seed - 9 mL silicic acid - 25 g zeolite powder. The following nursery conditions were provided : Early sowing, dense seeding in a glass house with mulching overnight and no artificial heating, which were the ideal conditions for determining the effect on the seed. The nursery plants were evaluated for Gibberella. fujikuroi infection or to determine the recovery to normal growth of infected nursery plants in the Wagner pot. Seedlings emerged 2-3 days earlier for the SCS than they did for the non-SCS control, while damping-off and bakanae disease incidence were remarkably reduced. Specifically, bakanae disease incidence in the SCS was limited to only 7.8% for 80 days after sowing, as compared to 91.6% of the non-SCS control. For the 45-days-old SCS nursery seedlings, the fresh weight was increased by 11% and was two times heavier, with only mild damage compared to that observed for non-SCS. Even after transplanting, SCS treatment contributed to a lower incidence of further infections and possibly to recovery of the seedlings to normal growth as compared to that observed in symptomatic plants in the pot. The active pathogenic macro-conidia and micro-conidia were considerably lower in the soil, root, and seedling sheath base of the SCS. In particular, the underdeveloped macro-conidia with straight oblong shape without intact septum were isolated in the SCS ; this phenotype is likely to be at a comparative etiological disadvantage when compared to that of typical active macro-conidia, which are slightly sickle-shaped with 3-7 intact septa. A active intact conidia with high inoculum potential were rarely observed in the tissue of the seedlings treated only in the SCS. We propose that promising result was likely achieved via inhibition of the development of intact pathogenic conidia, in concert with the aerobic, acidic conditions induced by the physiochemical characteristics associated with the air porosity of zeolite, alkalinity of silicate and the seed husk as a carbon source. In addition, the resistance of the healthy plants to pathogenic conidia was also important factor.

• Journal of Food Hygiene and Safety
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• v.35 no.1
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• pp.51-59
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• 2020

This study evalutated the risk of foodborne illness from Vibrio spp. (Vibrio vulnificus and Vibrio cholerae) through sea squirt consumption. The prevalence of V. vulnificus and V. cholerae in sea squirt was evaluated, and the predictive models to describe the kinetic behavior of the Vibrio in sea squirt were developed. Distribution temperatures and times were collected, and they were fitted to probabilistic distributions to determine the appropriate distributions. The raw data from the Korea National Health and Nutrition Examination Survey 2016 were used to estimate the consumption rates and amount of sea squirt. In the hazard characterization, the Beta-Poisson model for V. vulnificus and V. cholerae infection was used. With the collected data, a simulation model was prepared and it was run with @RISK to estimate probabilities of foodborne illness by pathogenic Vibrio spp. through sea squirt consumption. Among 101 sea squirt samples, there were no V. vulnificus positive samples, but V. cholerae was detected in one sample. The developed predictive models described the fates of Vibrio spp. in sea squirt during distribution and storage, appropriately shown as 0.815-0.907 of R² and 0.28 of RMSE. The consumption rate of sea squirt was 0.26%, and the daily consumption amount was 68.84 g per person. The Beta-Poisson model [P=1-(1+Dose/β)^-α] was selected as a dose-response model. With these data, a simulation model was developed, and the risks of V. vulnificus and V. cholerae foodborne illness from sea squirt consumption were 2.66×10^-15, and 1.02×10^-12, respectively. These results suggest that the risk of pathogenic Vibrio spp. in sea squirt could be considered low in Korea.

• Parasites, Hosts and Diseases
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• v.30 no.2
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• pp.101-112
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• 1992

The natural killer cell activity of splenocytes and TBC, active NK cells, recycling capacity of natural killer cells were observed by means of both the 51Cr-release cytotoxicity assay and single cell cytotoxicity assay against YAC-1. CSH/HeJ mice were infected intranasally with $1{\times}10^4{\;}or{\;}1{\times}10^5$ trophosoites of pathogenic Acanthamoeba culbertsoni. The infected mice showed mortality rate of 34% in $1{\times}10^4$ group and 65% in 1{\times}10^5 group, and mean survival time was $16.40{\pm}3.50$ {\;}and{\;}3.20{\pm}4.09$ days respectively. The cytotoxic activity of natural killer cells of the 2 groups was significantly higher than that of non-infected mice from the 12th hour to the 2nd day after infection, showing the highest on the first day. On the l0th day after infection, the cytotoxic activity of natural killer cells was significantly suppressed as compared with that of the control. There was no significant difference in NK cell cytotoxicity between two infected groups. The targetbinding capacity and active NK cells of natural killer cells in $1{\times}10^5$ trophosoite infected mice was significantly increased on the 12th hour and the first day after infection as compared with the control group. Maximal recycling capacity (MRC) was not changed during the observation period. The present results indicated that the elevation of natural killer cell activity in the mice infected with A. culbertsoni was due to elevation of target.binding capacity and increased active NK cells of natural killer cells, and not due to the maximal recycling capacity of the individual NK cell, and there was no difference between two experimental dose groups.

• Proceedings of the Korean Environmental Health Society Conference
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• 2005.06a
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• pp.45-72
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• 2005

C. neoformans-associated cryptococcosis is primarily a disease of immunocompromised persons, has a world-wide distribution, and is often spread by pigeons in the urban environment. In contrast, C. gattii causes infection in normal hosts, has only been described in tropical and semi-tropical areas of the world, and has a unique niche in river gum Eucalyptus trees. Cryptococcosis is acquired through inhalation of the yeast propagules from the environment. C. gattii has been identified as the cause of an emerging infectious disease centered on Vancouver Island, British Columbia, Canada. No cases of C. gattii-disease were diagnosed prior to 1999; the current incidence rate is 36 cases per million population. A search was initiated in 2001 to find the ecological niche of this basidiomycetous yeast. C. gaftii was found in the environment in treed areas of Vancouver Island. The highest percentage of colonized-tree clusters were found around central Vancouver Island, with decreasing rates of colonization to the north and south. Climate, soil and vegetation cover of this area, called the Coastal Douglas fir biogeoclimatic zone, is unique to British Columbia and Canada. The concentration of airborne C. gattii was highest in the dry summer months, and lowest during late fall, winter, and early spring, months which have heavy rainfall. The study of the emerging colonization of this organism and subsequent cases of environmentally acquired disease will be informative in planning public health management of new routes of exposure to exotic agents in areas impacted by changing climate and land use patterns. Cryptococcosis is an infection associated with an encapsulated, basidiomycetous yeast Cryptococcus neoformans. The route of entry for this organism is through the lungs, with possible systemic spread via the circulatory system to the brain and meninges. There are four cryptococcal serogroups associated with disease in humans and animals, distinguished by capsular polysaccharide antigens. Cryptococcus neoformans: variety grubii (serotype A), variety neoformans (serotype D), and variety gattii (serotypes B and C) (Franzot et at. 1999). C. neoformans variety gattii has recently been elevated to species status, C. gattii. C. neoformans val. grubii and var. neoformans have a world-wide distribution, and are particularly associated with soil and weathered bird droppings. In contrast, C. gattii (CG) is not associated with bird excrement, is primarily found in tropical and subtropical climates, and has a restricted environmental niche associated with specific tree species. (Ellis & Pfiffer 1990) Ellis and Pfeiffer theorize that, as a basidiomycete, CG requires an association with a tree in order to become pathogenic to mammals. In Australia, CG has been found to be associated with five species of Eucalypts, Eucalyptus camaldulensis, E. tereticornis, E. blakelyi, E. gomphocephala, and E. rudis. Eucalypts, although originally native to Australia, now have a world-wide distribution. CG has been found associated with imported eucalypts in India, California, Brazil, and Egypt. In addition, in Brazil and Columbia, where eucalypts have been naturalized, native trees have been shown to harbour CG (Callejas et al. 1998; Montenegro et al. 2000). In British Columbia, Canada, since the beginning of 1999, there have been 120 confirmed cases of cryptococcal mycoses associated with CG in humans, including 4 fatalities (data from British Columbia Centre for Disease Control), and over 200 cases in animal pets in BC (data from Central Laboratory for Veterinarians). What is remarkable about the BC outbreak of C. gattii-cryptococcosis is that all of the cases have been residents of, or visitors to, a narrow area along the eastern coast of Vancouver Island, BC, from the tip of the island in the south (Victoria) to Courtenay on the north-central island as illustrated in Figure 1. Of the first 38 human cases, 58% were male with a mean age of 59.7 years (range 20 - 82): 36 cases (95%) were Caucasian. Ten cases (26%) presented with meningitis, the remainder presented with respiratory symptoms. Cultures recovered from cases of cryptococcosis associated with the outbreak were typed as serogroup B, which is specific to CG (Bartlett et al. 2003). This was the first reported outbreak of CVG in Canada, or indeed, the world. Where infection with CG is endemic, for example, Australia, the incidence of cryptococcosis ranges from 1.8 - 4.7 per million between the southern and northern states (Sorrell 2001). However, the overall incidence of cryptococcosis in immunocompenent individuals has been estimated at 0.2 per million population per year (Kwon-Chung et al. 1984). The population of Vancouver Island is approximately 720,000,consequently, even if the organism were endemic, one would expect a maximum of 0.15 cases of cryptococcal disease annually.

• Parasites, Hosts and Diseases
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• v.26 no.1
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• pp.39-44
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• 1988

A pathogenic free-living amoeba, Naegleria fowleri, is a causative protozoan parasite of primary amebic meningoencephalitis in human and experimental animals. It is known that humoral and cellular immunity contribute as the defence mechanism of host against this organism. Recently splenectomy has been argued on its effect on host defence mechanisms. The present study was aimed to observe the enact of immunization in splenectomized mice. For immunization, $5~10{\times}10^5$ trophozoites of Naegleria fewleri o 359 were intraperitoneally inoculated once a week for two weeks to BALB/c mice, and $5~10{\times}10^4$ of ameba trophozoites were intranasally inoculated for infection after splenectomy and/or immunization. ELISA technique was applied for the detection of seum IgG antibody levels. Experimental animals were divided into 4 groups; I. splenectomized and immuniEed; ll. splenectomized only; III. immunized only; IV. not splenectomized nor immunized. The results obtained were as follows: 1. Mortality rates of splenectomized and immunized mice in group I (38.1%) and immunized only in group III (25.0%) were lower than those of not immunized mice in group II (50%) and control group, IV (46.4%). 2. Survival times of mice in group I, II, III and IV were $20.1{\pm}3.6$, $17.3{\pm}4.5$, $20.4{\pm}7.0$ and $19.6{\pm}7.6$ days respectively, and there were no significant differences between them. 3. ELISA values (absorbance at 492nm) of group I (1, $10{\pm}0.29$) and group III ($1.31{\pm}0.28$) were significantly higher than that of group IV($0.24{\pm}0.37$) at day 31 of infection (p<0.05). Conclusively, it is presumed that humoral immunity against N. fowleri may operate as ever, after immunization, even though the mouse was splenectomized.

• Journal of Food Hygiene and Safety
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• v.24 no.2
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• pp.174-181
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• 2009

The consumption of "ready-to-eat" agricultural products is recently increasing and the safety of these agricultural products is forefront of public concerns. The 120 samples of paprikas, strawberries and tomatoes, which are the representative exported agricultural products, were purchased at the department stores and discount stores in Daejeon. And we determined the microbiological and parasitological contamination level of these agricultural products using culture media, multiplex PCR, commercial bacterial detection kit and microscopy, and also evaluated the decontamination method. Mean counts of total aerobic bacteria from these agricultural products ranged from $1.3{\times}10^4$ CFU/g to $1.8{\times}10^5$ CFU/g, and mean counts of coliforms ranged from $1.4{\times}10^3$ CFU/g to $9.6{\times}10^3$ CFU/g. There was no significant difference in the level of bacterial contamination between the agricultural products from department stores and the ones from discount stores. Strawberry showed the highest contamination level for the bacteria and we also found the unidentified parasite eggs. Enterobacter cloacae was the most frequently isolated bacteria strain, but no food poisoning pathogenic bacteria except Staphylococcus aureus was isolated from the products by multiplex PCR. Compared to unwashed products, tab water-washed ones showed 80% decrease of the counts of total aerobic bacteria on the agricultural products, and the rates decreased more by incorporating detergent or ultrasonic wave treatment. We concluded that the biological contamination levels among paprikas, strawberries and tomatoes were the highest in strawberries, but there were not significant difference according to distribution systems.