• Korean Journal of Microbiology
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• v.51 no.4
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• pp.364-373
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• 2015

As an effort to utilize alginate, 103 bacterial isolates that were positive for the alginate lyase activity were isolated from various clams and seawater samples collected in Incheon coastal area. Among them, 3 strains (M1-2-1, M6-1, and C8-15) were finally selected for further analysis based on their activities at higher levels than others. These isolates were all Gram-negative and rod shaped halophilic bacteria with motility. According to their physiological and biochemical properties as well as DNA sequence of their 16S rRNA genes, M1-2-1 and M6-1 were identified as a member of genus Pseudoalteromonas and C8-15 belonged to genus Vibrio. They exhibited the alginate degrading activity at the maximal level when they were cultured in APY broth for 6-8 h at $25^{\circ}C$. Both their growth and the enzyme activity were greatly enhanced when NaCl was added to the growth medium. The crude alginate lyases from the supernatants of the bacterial cultures showed the highest activity at $45^{\circ}C$ and pH 7.0-8.0. M1-2-1 and M6-1 produced 2.723 and 1.976 g/L of reducing sugar from alginate, respectively, suggesting that they have potential for commercial application.

• 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.

• Applied Biological Chemistry
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• v.9
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• pp.9-27
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• 1968

Studies were carried out in order to elucidate chemical components and microflora in three types of soy-sauce, 12-year aged soy-sauce prepared by improved method. 7-year aged and 20-year aged soy-sauce prepared by ordinary method. They results are summarized as follows: 1. The followings are found to be the important factors affecting the quality of soy sauce. a. Organic acids, reducing sugars and free amino acids were increased in the course of storage. b. In the aged soy-sauces under study non-volatile organic acid increased while volatile organic acid decreased and the total acidity was dependent only upon tie latter. c. It was found that suit concentration decreased during the storage. 2. The results of investigation of microflora in the stored soy-sauce are shown as follows. Soy-sauce Improved Ordinary Microbe 12-Y. 20-Y. 7-Y. Aerobic bacteria colony/1ml. 6 123 2 Halophilic lactic acid bacteria colony/1ml. 4 6 10 Osmophilic yeast colony/1ml. $828{\times}10^4$ 248 - b. In the stored soy-sauces, aerobic bacteria are incapable of growing due to drop in pH value and the influence of salt concentration. c. Halophilic lactic acid bacteria are incapable of growing due to drop in pH value, even the salt concentrations decreased during the storage. d. Osmophilic yeast are still growing in low pH value and in the decreasing salt concentration during the strage. 3. The results of amino acid analysis by paper partition chromatographic and calorimetric methods are shown as follows. a. Fourteen kinds of amino acid and thirteen amino acids were detected in the soy-sauce of 12-year aged improved soy-sauce and 7-year aged and 20-year aged ordinary one, respectively. b. The contents of aspartic acid, glutamic acid, serine, valine, leucine, lysine, histidine and methionine increased in the 20-year aged ordinary soy-sauce compared to the 7-year aged one. On the other hand those of alanine, tyrosine, phenylalanine and cystine decreased. 4. The results of sugar analysis by paper chromatography are as follows. a. In the 12-year aged improved soy-sauce, galactose, glucose, arabinose, xylose, rhamnose, maltose and an unknown were detected, and their amounts were in the above order except maltose and an unknown. b. Both in the 7 and 12-year aged ordinary soy-sauces, galactose, arabinose, xylose, glucose and rhamnose were detected and the amounts of the sugars were in the above order. c. In the non-aged ordinary soy-sauce, glucose was not detected but detected from 7-year and 20-year aged ordinary soy-sauce. 5. The results of organic acid analysis by paper chromatography were as follows. a. As volatile acids, acetic, propionic and butyric acids were detected in the 7-year aged ordinary soy-sauce. On the other hand in both the 20-year aged ordinary soy-sauce and the 12-year aged improved ones, only acetic acid was abundant while propionic and butyric acids were round in trace. It was found that propionic and butyric acids, as the unpleasant flavor components, decreased during the storage. b. In the ordinary soy-sauce, citric acid were produced during the storage and lack, malic and tartaric acids increased in the course of aging while succinic, glycolic, fumaric and malonic acids were shown to decrease. Glutaric and oxalic acids disappeared. Citric acid was produced also in the improved soy-sauce, but lactic, tartaric, succinic, malic, and glycolic acids decreased, while both malonic and glutaric acids disappeared. From the above results the citric acid production was considered to be a favorable factor for the taste. c. In the aged soy-sauces, pyruvic, α-ketoglutaric and probably acetoacetic and oxaloacetic acids (both in trace) were present and their amounts were in the above order. All of the α-keto acid abruptly decreased during the storage.

• Korean Journal of Microbiology
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• v.45 no.2
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• pp.193-199
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• 2009

A bacterium producing the halotolerant extracellular protease was isolated from squid jeotgal, and was identified as Bacillus sp. SJ-10 based on morphological, physiological and biochemical characteristics, as well as phylogenetic analysis using 16S rRNA gene sequence. The strain grew at $20^{\circ}C\sim55^{\circ}C$, pH 5~8, and 0%~14% NaCl and optimal growth conditions were $35{\pm}5^{\circ}C$, pH 7, and 5% NaCl. The major cellular fatty acids were anteiso-$C_{15:0}$, anteiso-$C_{17:0}$, and $C_{16:0}$ DNA G+C content was 50.58 mol% and menaquinone consisted of MK-7 Phylogenic analysis based on the 16S rRNA gene sequence indicated that SJ-10T belongs to the genus Bacillus. About 40 kDa of the salt-tolerant protease was purified by 40% ammonium sulfate saturation and Mono Q column chromatography. The optimal activity of the protease was pH 8 and stable at pH 5~10. The optimum temperature and NaCl concentration were $35{\pm}5^{\circ}C$ and $5{\pm}1%$, respectively.

• Journal of Microbiology and Biotechnology
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• v.29 no.7
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• pp.1104-1116
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• 2019

In this study, we investigated the potential of using sediment bioelectrochemical systems (SBESs) for in situ treatment of the water and sediment in brackish aquaculture ponds polluted with uneaten feed. An SBES integrated into a laboratory-scale tank simulating a brackish aquaculture pond was established. This test tank and the control (not containing the SBES) were fed with shrimp feed in a scheme that mimics a situation where 50% of feed is uneaten. After the SBES was inoculated with microbial sources from actual shrimp pond sediments, electricity generation was well observed from the first experimental week, indicating successful enrichment of electrochemically active bacteria in the test tank sediment. The electricity generation became steady after 3 weeks of operation, with an average current density of $2.3mA/m^2$ anode surface and an average power density of $0.05mW/m^2$ anode surface. The SBES removed 20-30% more COD of the tank water, compared to the control. After 1 year, the SBES also reduced the amount of sediment in the tank by 40% and thus could remove approximately 40% more COD and approximately 52% more nitrogen from the sediment, compared to the control. Insignificant amounts of nitrite and nitrate were detected, suggesting complete removal of nitrogen by the system. PCR-DGGE-based analyses revealed the dominant presence of Methylophilus rhizosphaerae, Desulfatitalea tepidiphila and Thiothrix eikelboomii, which have not been found in bioelectrochemical systems before, in the bacterial community in the sediment of the SBES-containing tank. The results of this research demonstrate the potential application of SBESs in helping to reduce water pollution threats, fish and shrimp disease risks, and thus farmers' losses.

• 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 $7.2{\times}10^1{\sim}9.9{\times}10^2\;CFU/100ml$.

• Food Science and Preservation
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• v.20 no.6
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• pp.791-797
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• 2013

This study was conducted to investigate the changes in the composition and microbiological properties of domestic sun-dried salt (white and gray salts) according to their collection time and production method. The results showed that the moisture contents of the white and gray sun-dried salts were 10.4~13.2% and 5.2~8.0%, respectively, and the sand contents were 0.1% and 0.2~0.3%, respectively, according to the month. Several samples exceeded the criteria of 15% moisture content and 0.2% sand content. The ash content and salinity of gray salt (below 85% and 90%, respectively) were higher than those of white salt (both below 80%). The total chloride contents of the salts collected in September and October were slightly lower than that of the others and exceeded the criteria of above 40%. In the case of mineral contents, there was no significant difference among the collection times because the analyses showed a marked deviation. The microbiological analysis showed that there was no significant difference among the production method, but the salt samples collected in September and October had relatively high detection rates of total aerobe, staphylococci, and halophilic bacteria.

• 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.

• 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.