• Korean Journal of Fisheries and Aquatic Sciences
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• v.19 no.2
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• pp.118-126
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• 1986

Vibrio vulnificus is a recently recognized halophilic organism that nay cause serious human infections. Patients infected with V. vulnificus often have a history of exposure to the sea, suggesting that the organism may be common inhabitant of marine environment. The purpose of this experiment is to investigate the distribution and bacteriological characteristics of V. vulnificus. The strain used in this experiment was isolated from sea water and sea products such as common octopus (Octopus variabilis), ark shell (Anadara broughtonii), blue crab (Ericheir japonica), and sea squirt (Synthia roretzi) collected in Pusan area from July to October in 1985. V. vulnificus was frequently isolated in August when temperature of sea water was around $26^{\circ}C$ and rarely isolated in October when temperature of sea water was around $18.5^{\circ}C$. The distinctive biochemical characteristics of V. vulnificus were ONPG hydrolysis positive and fermented lactose and not grown in peptone water contained $8\%$ NaCl. The optical density at 660 nm of the growth of V. vulnificus was reached maximum level after 8 hours of culture at $35^{\circ}C$ in brain heart infusion broth but that of V. vulnificus was little increased at $15^{\circ}C$ for 14 hours. Optimum temperature and pH for the growth of V. vulnificus were around $35^{\circ}C$ and 8.0. The specific growth rate and the generation time of V. vulnificus isolated from the samples were $1.21\;hr^{-1}$, 34 min at $35^{\circ}C$ and $0.61\;hr^{-1}$, 69 min at $25^{\circ}C$, respectively. V. vulnificus did not grow on eosin-methylene-blue agar, salmonella-shigella agar, deoxycholate agar but grew well on Endo agar, xylose-lysine-deoxycholate agar and hektoen enteric agar. On Endo agar, the colonies of V. vulnificus were red and achieved a diameter of 2 to 4 mm as a feature enabling differentiation of V. vulnificus from other Vibrio spp. V. vulnificus grow well on TCBS agar forming green colonies. V. vulnificus refrigerated at $4^{\circ}C$ exhibited a linear decline of its viablity as 1 log cycle in every 16 hours storage, while V. vulnificus freezed at $-18^{\circ}C$ almost became extinct.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.7
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• pp.1115-1124
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• 2013

The present study was conducted to ensure the diversity of domestic solar salt by analyzing the composition and microbiological characteristics of solar salt (from Docho island: DS) and the flower of salt produced in different Korean salt flats (Sinui island: SF, Bigum island: BF, and Docho island: DF). The analyses showed that the moisture content of the three types of flower of salt and solar salt ranged from 10.54~13.82% and NaCl content ranged from 78.81~84.61%. The mineral content of those salts ranged from 3.57~5.51%. The content of insoluble matter in these salts was $0.01{\pm}0.00{\sim}0.05{\pm}0.00%$. The sand content of these salts was $0.01{\pm}0.01{\sim}0.03{\pm}0.01%$. By Hunter's color value analysis, the color of the flower of salt was brighter and whiter than solar salt. The salinity of the flower of salt was a little higher than solar salt as well. The magnesium and potassium ion content of DF was $9,886.72{\pm}104.78mg/kg$ and $2,975.23{\pm}79.73mg/kg$, respectively, which was lower than the content in SF, BF, and DS. The heavy metal content of all salts was acceptable under the Korean Food Sanitation Law. The flower of salt was confirmed to be sweeter and preferable to solar salt. More than 80% of the solar salt crystals were 2~3 mm in size, whereas crystals from the flower of salt were 0.5~2 mm in size. The bacterial diversity of DF and DS were investigated by culture and denaturing gradient gel electrophoresis (DGGE) methods. The number of cultured bacteria in flower of salt was approximately three times more than solar salt. By DGGE analysis, major microbes of DF were Maritimibacter sp., Cupriavidus sp., and unculturable bacteria, and those of DS were Cupriavidus sp., Dunalidella salina and unculturable bacteria. The results of DGGE analysis showed that major microorganisms in solar salts were composed of unidentified and unculturable bacteria and only a few microorganisms were culturable.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.9 no.2
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• pp.79-110
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• 1976

In Korea fermented fish and shellfish have traditionally been favored and consumed as seasonings or further processed for fish sauce. Three major items in production quantity among more than thirty kinds which are presently available in the market are fermented anchovy, oyster and small shrimp. They are usually used as a seasoning mixture of Kimchi in order to provide a distinctive flavor. Fermented small shrimp, Acetes chinensis is most widely and largely used ana occupies an important position in food industry of this country. But no study on its taste compounds has been reported. This study was attempted to establish the basic data for evaluating taste compounds of fermented small shrimp. The changes of such compounds during fermentation as free amino acids, nucleotides and their related compounds, TMAO, TMA, and betaine were analysed. In addition, change in microflora during the fermentation under the halophilic circumstance was also investigated. The samples were prepared with three different salt contents of 20, 30 and $40\%$ to obtain the proper degree of fermentation at a controlled tempeature of $20{\pm}2^{\circ}C$. The results are summarized as follows: Volatile basic nitrogen increased rapidly until 108 days of fermentation and afterwards it tended to increase slowly. Amino nitrogen also increased rapidly until 43 days of fermentation and then increased slowly. Extract nitrogen increased and marked the maximum value at 72 day fermentation and then decreased slowly. ADP, AMP and IMP tended to degrade rapidly while hypoxanthine increased remarkably at 27 day fermentation but slightly decreased at 72 day fermentation. It is presumed that the characteristic flavor of fermented small shrimp might be attributed to the relatively higher content of hypoxanthine. In the free amino acid composition of fresh small shrimp abundant amino acids were proline, arginine, alanine, glycine, lysine, glutamic acid, leucine, valine and threonine in order. Such amino acids like serine, methionine, isoleucine, phenylalanine, aspartic acid, tyrosine and histidine were poor. In small shrimp extract, proline, arginine, alanine, glycine, lysine and glutamic acid were dominant holding $18.5\%,\;14.6\%,\;10.8\%,\;8.7\%,\;8.1\%\;and\;7.7\%$ of total free amino acids respectively. The total free amino acid nitrogen in fresh small shrimp was $63.9\%$ of its extract nitrogen. The change of free amino acid composition in the extract of small shrimp during fermentation was not observed. Lysine, alanine glutamic acid, proline, glycine and leucine were abundant in both fresh sample and fermented products. The increase of total free amino acids during 72 day fermentation reached approximately more than 2 times as compared with that of fresh sample and then decreased slowly. Fermented small shrimp with $40\%$ of salt was too salty to be commercial quality as the results of organoleptic test showed. It is found that 72 day fermentation with $20\%\;and\;30\%$ of salt gave the most favorable flavor. It is convinced that the characteristic flavor of fermented small shrimp was also attributed to such amino acids as lysine, proline, alanine, glycine and serine known as sweet compounds, as glutamic acid with meaty taste, and as leucine known as bitter taste. The amount of betaine increased during fermentation and reached the maximum at 72 day fermentation and then decreased slowly TMA increased while TMAO decreased during fermentation. The amount of TMAO nitrogen in fermented small shrimp was $200mg\%$ on moisture and salt free base. Betaine and TMAO known as sweet compounds were abundant in fermented small shrimp. It is supposed that these compounds could also play a role as important taste compounds of fermented small shrimp. At the initial stage of fermentation, Achromobacter, Pseudomonas, Micrococcus denitrificans which belong to marine bacteria were isolated. After 40 day fermentation, they disappeared rapidly while Halabacterium, Pediococcus, Sarcian, Micrococcus morrhuae and the yeasts such as Saccharomyces sp. and Torulopsis sp. dominated. It is concluded that the most important taste compounds of fermented small shrimp were amino acids such as lysine, proline, alanine, glycine, serine, glutamic acid, and leucine, betaine, TMAO and hypoxanthine.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.7 no.3
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• pp.105-114
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• 1974

Identification and change of microflora during the fermentation of anchovy Engraulis japonica, under the halophilic circumstance were investigated. The change of salinity and pH in meat and juice which decide the environment for microorganism and decomposition of nitrogenous compound which functions as a nutrient source were also discussed by measuring the content of total-N, amino-N, nonprotein-N, TMA and VBN, The fresh anchovy was mixed with rock salt (20 percent w/w) and stocked for six months. Through the fermentation lag phase of viable cells extended for 20 days that was obviously larger compared with other circumstances, hereafter increased to reach the maximum value of $5\times10^4$ total count per gram at 35 day stock. The stationary phase proceeded for 25 days. 540 strains were isolated and among them 11 genus of bacteria, 3 genus of yeasts, were identified and other 2 yeast strains of unidentified. At the initial stage of fermentation, Pseudomonas, and Helobacterium prevalently grew, at the middle stage, they disappeared rapidly and Pediococcus and yeasts completely dominated, where they are assumed to get directly involved with fermentation of fish, The PH value tended to decrease in the progress of fermentation and at 100 day stock it showed the minimum value of 5.5 to 5.6 in both meat and juice. The highest salinity of meat decreased to 18 percent, while in juice it decreased to 28 percent since 50 days stock. The content of total-N in meat gradually decreased to 2.8 percent, while in juice it increased to 2.3 percent at 100 day stock, However nonprotein-N was 1.8 percent and amino-N was 1.1 Percent. Since 100 days stock, the increasing rate of amino-M is too low it could be judged to entered the final stage of fermentation, In the first 20 days stock, the increase of VBN and TMA can be explained by the growth of putrefactive bacteria such as pseudomonas on the meat before salts penetrate into the fish meat, while reincrement after 100 days stock, is explained by decomposition of free amino acid due to the reactions of bacteria and enzymes.

• Korean Journal of Food Science and Technology
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• v.25 no.5
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• pp.449-455
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• 1993

Vibrio vulnificus is a recentry recognized halophilic organism that may cause human infections. Patients infected with Vibrio vulnificus often have a history of exposure to the sea, suggesting that the organism may be common inhabitant of marine environment. In this studies 314 samples were collected from 9 sites of Chonnam coastal area, from April to Jun, 1993. Isolation rates of V. vulnificus were 7.1% in total samples, 33.3% in sea water, 55.5% in sediment, 1.7% in fish and shellfish, 6.3% in kitchen environment. In each areas of Chunnam V. vunificus were isolated from 3.6% to 11.4%. The isolation rates of V. vulnificus was correlated positively with organic matter and COD in sea warer and sediment. In sea water, Microorganism population were $4.5{\times}10^3{\sim}3.5{\times}10^5\;CFU/ml$, Vibrio strains were $1.8{\times}10^l4.6{\times}10^3\;CFU/ml$, Coliform bacteria were $1.9{\times}10^1{\times}3.7{\times}10^2\;CFU/100ml$. In sediment, Microorganism population were $4.8{\times}10^3{\sim}5.2{\times}10^5\;CFU/ml$. Vibrio strains were $1.9{\times}10^2{\sim}8.4{\times}10^4\;CFU/ml$, Coliform bacteria were $7.2{\times}10^1{\sim}9.9{\times}10^2\;CFU/100ml$.

• Korean Journal of Food Science and Technology
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• v.30 no.4
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• pp.908-915
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• 1998

By using Korean native soybean, traditional meju was prepared in Chuncheon, Kangweondo according to the traditional process. Analysis of physico-chemical, enzymatic and microbiological changes during meju fermentation were carried out in order to obtain a basic information for industrial scale production of meju. The enviroments for natural meju fermentation were $10{\sim}15^{\circ}C$ and $60{\sim}70%{\;}RH$. Moisture content decreased from 59% to 11% (exterior section) and 19% (interior section). the pH of meju rapidly increased up to 8.5 at $33^{rd}{\;}day$ of fermentation and thereafter decreased down to 7.9 at $70^{th}{\;}day$ of fermentation. Souble protein content was 1.47% at initial stage and increased up to $6.31{\sim}7.34%$ at $33^{rd}{\;}day$ of fermentation. Amino nitrogen content was $460{\sim}770{\;}mg%$ at $70^{th}{\;}day$ of fermentation. the color of meju became gradually black and decreased in redness and yellowness. During the process, protease and lipase seemed to play an important role in the digestion of soy protein and fat. Acidic protease activity increased up to $135.9{\sim}152.4{\;}unit/g$ at $33^{rd}{\;}day$ of fermentation and were $181.3{\sim}272.6{\;}unit/g$ at $70^{th}{\;}day$ of fermentation. Lipase activity increased up to 6 unit/g (interior section) and 15 unit/g (exterior section) at $70^{th}{\;}day$ of fermentation. the viable cell count of meju was at the level of $10^8{\;}CFU/g$ during the overall fermentation period. Aerobic halophilic count was $1.51{\times}10^7{\;}CFU/g$ at initial stage and maintained $10^8{\;}CFU/g$ level during the process. Initial anaerobic cell count was $2.0^9{\times}10^4{\;}CFU/g$ and increased up to $10^5{\;}CFU/g$ level at 47 days. Yeast and mold counts were $10^4{\sim}10^5{\;}CFU/g$ for the fermentation period.

• Journal of Food Hygiene and Safety
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• v.21 no.4
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• pp.213-217
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• 2006

The halophilic bacterium Vibrio vulnificus is known to be a foodborne pathogen that causes septicemia in human. V. vulnificus infection is characterized by the high fatality rates and the primary attack against a person who have underlying diseases such as liver cirrhosis. However, there is no effective treatment for V. vulnificus septicemia except for classical treatments such as antibiotics. Recently, it has been known that lipoprotein (LDL) plays a major role in the protection against infection and inflammation. Consequently in this paper we analyzed the effects of LDL on V. vulnificus septicemia. We purified V. vulnificus cytolysin, a major virulent factor of V. vulnificus infection and measured inhibitory effects of mouse serum, cholesterol, and LDL on its hemolytic activity. Next experiments were performed to investigate whether LDL has a protective role against septicemia induced by V. vulnificus in mice. Intraperitoneal injection of LDL (1mg as protein) into mice 3hr before V. vulnificus $(1\times10^6\;CFU)$ injection, and V. vulnificus -induced lethality was determined. For the determination the relationship between LDL or cholesterol and prognosis, we determined serum levels of cholesterol and lipoprotein from V. vulnificus septicemia patients (n=15) who had visited the Chonbuk National University Hospital in Chonju. V. vulnificus cytolysin -induced hemolysis of mice erythrocytes was completely inhibited by serum, cholesterol, and low-density lipoprotein. V. vulnificus- induced lethality of mice injected with LDL showed only 40% compared to 100% of control. In survival groups (n=4) of V. vulnificus septicemia patients (n=15), their serum LDL and cholesterol revealed normal levels ($153.3{\pm}40.7,\;LDL;\;190.8{\pm}16.3$, Total cholesterol). However, in death groups (n=11) showed very low levels ($35.6{\pm}13.9,\;LDL;\;59.2{\pm}15.1$, Total cholesterol). Our study indicates that cholesterol and LDL are a prognosis indicator of V. vulnificus septicemia as well as an inhibitor of virulent action of V. vulnificus cytolysin. We suggested that the serum levels of cholesterol or LDL would be major index in the treatment and prevention of V. vulnificus septicemia.

• Journal of Food Hygiene and Safety
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• v.30 no.3
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• pp.281-288
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• 2015

Vibrio is a genus of Gram-negative, curved, halophilic, and non-spore-forming bacteria. Some of the Vibrio species, such as V. cholerae and V. parahaemolyticus, often contaminate seafood products and occasionally cause human diseases when the seafood products are ingested. A total of 24 Vibrio strains were isolated from shrimp samples on Thiosulphate citrate bile salt sucrose (TCBS) media in this study. All of the 24 isolates were confirmed to belong to the genus Vibrio by using 16S rRNA gene sequence analyses. Vitek 2 system and species-specific polymerase chain reaction (PCR) methods were used to further identify a total of 29 Vibrio strains at the species level, including the 24 shrimp Vibrio isolates and five Vibrio reference strains. The specificities of the two methods to identify Vibrio strains at the species level were compared in this study. The species-specific PCR method was designed to detect five different Vibrio species, such as Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio alginolyticus, and Vibrio mimicus. From the 24 Vibrio shrimp isolates, the Vitek 2 system method could identify 15 (62.5%) strains as Vibrio species and 7 (29.2%) strains as non-Vibrio species, but could not identify the rest 2 (8.3%) strains. But species-specific PCR method could identify 16 (66.7%) strains as Vibrio species and could not identify the rest 8 (33.3%) strains. Among the 24 Vibrio shrimp strains, these two methods could unanimously identify 7 (7/24, 29.2%) strains (2 V. parahaemolyticus, 4 V. alginolyticus, and 1 V. mimicus). Considering that such different identification results were obtained using the two different methods in this study, identification method for Vibrio species must be carefully chosen.

• Journal of Microbiology and Biotechnology
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• v.29 no.10
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• pp.1607-1623
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• 2019

Sediment bioelectrochemical systems (SBESs) can be integrated into brackish aquaculture ponds for in-situ bioremediation of the pond water and sediment. Such an in-situ system offers advantages including reduced treatment cost, reusability and simple handling. In order to realize such an application potential of the SBES, in this laboratory-scale study we investigated the effect of several controllable and uncontrollable operational factors on the in-situ bioremediation performance of a tank model of a brackish aquaculture pond, into which a SBES was integrated, in comparison with a natural degradation control model. The performance was evaluated in terms of electricity generation by the SBES, Chemical oxygen demand (COD) removal and nitrogen removal of both the tank water and the tank sediment. Real-life conditions of the operational parameters were also experimented to understand the most close-to-practice responses of the system to their changes. Predictable effects of controllable parameters including external resistance and electrode spacing, similar to those reported previously for the BESs, were shown by the results but exceptions were observed. Accordingly, while increasing the electrode spacing reduced the current densities but generally improved COD and nitrogen removal, increasing the external resistance could result in decreased COD removal but also increased nitrogen removal and decreased current densities. However, maximum electricity generation and COD removal efficiency difference of the SBES (versus the control) could be reached with an external resistance of $100{\Omega}$, not with the lowest one of $10{\Omega}$. The effects of uncontrollable parameters such as ambient temperature, salinity and pH of the pond (tank) water were rather unpredictable. Temperatures higher than $35^{\circ}C$ seemed to have more accelaration effect on natural degradation than on bioelectrochemical processes. Changing salinity seriously changed the electricity generation but did not clearly affect the bioremediation performance of the SBES, although at 2.5% salinity the SBES displayed a significantly more efficient removal of nitrogen in the water, compared to the control. Variation of pH to practically extreme levels (5.5 and 8.8) led to increased electricity generations but poorer performances of the SBES (vs. the control) in removing COD and nitrogen. Altogether, the results suggest some distinct responses of the SBES under brackish conditions and imply that COD removal and nitrogen removal in the system are not completely linked to bioelectrochemical processes but electrochemically enriched bacteria can still perform non-bioelectrochemical COD and nitrogen removals more efficiently than natural ones. The results confirm the application potential of the SBES in brackish aquaculture bioremediation and help propose efficient practices to warrant the success of such application in real-life scenarios.

• The Korean Journal of Mycology
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• v.49 no.1
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• pp.87-100
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• 2021

This study aimed to isolate wild yeasts obtained from flowers and soil of the Wolpyung park, Daejeon city and Gykpo beach, Buan, Jeollabuk-do in Korea, and to further characterize previously unrecorded wild yeast strains. In total, 88 strains of 62 different species of wild yeasts were isolated from 75 samples obtained from the Wolpyung park. Among these, six strains of Trichosporon moniliiforme and four strains each of Papiliotrema flavescens and Candida melibiosica were isolated. Additionally, 39 strains of 30 different species of wild yeasts were isolated from 35 samples collected from the Gykpo beach. Among the 127 isolated wild yeast strains, 10 strains, including Apiotrichum porosum ASCM32-1, were previously unrecorded. All the 10 previously unrecorded yeasts were oval or global in shape, and three strains, including Candida athensensis WP4-90-3, formed spores. Three strains, including Vishniacozyma taibaiensis WP13-2, were halophilic yeasts which grew in 15% NaCl-containing YPD(yeast extract-peptone-dextrose) medium. Five strains, including C. athensensis WP4-90-3, showed 15% ethanol resistance. Cell-free extracts from Candida oleophila WP5-19-1 and Wickerhamomyces anomalus HO9-2 showed the highest β-glucuronidase inhibitory activity (49.0%) and neutrophil elastase inhibitory activity (38.4%), respectively.