• Korean Journal of Fisheries and Aquatic Sciences
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• 제7권3호
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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.

• Journal of fish pathology
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• 제18권1호
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• pp.39-48
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• 2005

Pathogenicity of Vibrio tapetis, the causative bacterium of 'brown ring disease (BRD)' was evaluated in Manila clams (Ruditapes philippinarumi by artificially 0.1 $m\ell$ infection of $1.0\times10^5$cells and $1.0\times10^8$ cells at 20 $^{\circ}C$. A PCR assay based on 16S rRNA to detect the bacteria in clam tissues was established. Accumulative mortality of clams infected with $1.0\times10^7$cells and $1.0\times10^4$ cells per an individual of the bacteria was 67.5% and 7.5%, respectively. However, the deposit of brown pigment in the inner shells by accumulation of chonchiolin was not found. The bacteria were not be able to re-isolate from the infected clams by the conventional agar plate method but were easily detected by PCR assay established in this experiment. In clams artificially infected with 10 species of Vibrio, a 414bp for V. tapetis was detected in PCR assay. The specific band in the clams infected with $1.0\times10^4$cells per an individual of V. tapetis was detected only in gills one day after the infection but never be found in any tissues including gills three days after the infection. In the case of clams infected with $1.0\times10^8$cells per an individual of V. tapetis the specific band was detected in gills and intestine one day after the infection, in all tissues three days after the infection, and then in gills and adductor muscle nine days after the infection. The PCR assay was applied to detect V. tapetis in manila clam, surf clam (Mactra veneriformis), oyster (Crassostrea gigas) and Thomas' rapa whelk (Rapana venosa) taken from Taean and Gochang from April to July 2004. The infection rates were detected to 23.1% and 9.4% in the oyster and surf clam, while manila clam and Thomas' rapa whelk were not found.

• Journal of fish pathology
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• 제33권2호
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• pp.111-118
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• 2020

The genus Edwardsiella belonging to the family Enterobacteriaceae is a member of Gram-negative rod-shaped bacteria that cause disease in diverse aquatic organisms such as fish, amphibians and reptiles as well as avians and mammals including human throughout the world. This genus had been composed of three species, E. hoshinae, E. ictaluri and E. tarda, but recent researches erected two novel species, E. anguillarum and E. piscicida that were conventionally identified as E. tarda. In this study, we isolated seven strains belonging to the genus Edwardsiella from freshwater fishes that had been reared at inland fish farms in South Korea and investigated their biochemical characteristics and molecular phylogenetic relationships. The seven strains showed typical characteristics of four Edwardsiella species, E. anguillarum, E. ictaluri, E. piscicida and E. tarda, by biochemical analyses of Gram staining, indole and hydrogen sulfide (H2S) production, and API (Analytic Profile Index) 20E test. Molecular phylogenetic analyses inferred from DNA sequence data of both 16S ribosomal RNA (rRNA) and DNA gyrase subunit B (gyrB) genes were congruent with the biochemical characteristics. As a result, both biochemical and molecular phylogenetic analyses identified four strains isolated from three Anguilla species as E. anguillarum, E. piscicida and E. tarda, two strains from Pelteobagrus fulvidraco and Silurus asotus as E. ictaluri, and one strain from Moroco oxycephalus as E. piscicida. In this study, we isolated and successfully identified recently newly erected species, E. anguillarum and E. piscicida in addition to historically notorious pathogenic species, E. ictaluri and E. tarda. In the future study, systematic and comprehensive monitoring of the four Edwardsiella species are required for studying differences in pathogenicity among freshwater fishes.

• Journal of Life Science
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• 제28권2호
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• pp.223-228
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• 2018

Lactic acid bacteria were isolated from a variety of fermented seafoods and sea creatures from the East Sea Rim, Korea and were screened for ${\gamma}-amino$ butyric acid-producing (GABA) activity. Through a 16S rRNA sequence analysis, the bacteria of interest, which were GABA-positive on the thin-layer chromatography analysis, were recognized as three isolates of Lactobacillus (Lb.) brevis and one isolate of Lactococcus (Lc.) lactis. Lb. brevis FSFL0004 and FSFL0005 were isolated from fermented anglerfish and Lb. brevis FSFL0036 was derived from salted cutlass fish. The Lc. lactis strain FGL0007 was isolated from the gut of a brown sole flounder. According to HPLC analysis, the GABA contents produced by FSFL0004, FSFL0005, FSFL0036 and FGL0007 were equivalent to $10,754.37{\mu}g/ml$, $13,082.79{\mu}g/ml$, $12,290.19{\mu}g/ml$, and $45.07{\mu}g/ml$ respectively in 1% monosodium glutamate-supplemented methionyl-tRNA synthetase (MRS) broth. The four strains were inoculated in skim milk with 1% monosodium glutamate to commercialize the strains as starter cultures for GABA-enriched dairy products, and TLC results displayed the production of ${\gamma}-amino$ butyric acid by all four strains in the adaptation media. Lc. lactis FGL0007 demonstrated the greatest GABA production ($431.42{\mu}g/ml$) by HPLC analysis. The GABA production by lactic acid bacteria strains in the skim milk demonstrated in the present study may be helpful for the production of GABA-enriched dairy products.

• Korean Journal of Fisheries and Aquatic Sciences
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• 제30권3호
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• pp.319-328
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• 1997

To develop a beneficial microbial feed for the cultivation of rotifer, Brachionus plicatilis, an aerobic photosynthetic bacterium, Erythrobacter sp. $S\;\pi-I$ was isolated from marine structure at Haeundae beach in Pusan, Korea. Feeding effects of Erythrobacter sp. $S\;\pi-I$ on the growth of rotifer were analyzed comparing to other feeds such as PSB (purple nonsulfur bacteria), Chlorella sp. and baker's yeast. Erythrobacter sp. $S\;\pi-I$ contained more linoleic acid $(C_{18:3\omega3})$ and oleic acid $(C_{18:1\omega9})$ and amino acids than PSB (purple nonsulfur bacteria), Chlorella sp. and baker's yeast. The rotifer fed on Erythrobacter sp. $S\;\pi-I$ showed better effects than those fed on other feeds in the individual growth, size and weight. Also, the rotifer especially contained more eicosapentaenoic acid $(C_{20:5\omega3})$ and docosahexaenoic acid $(C_{22:6\omega3})$ in case of Erythrobacter sp. $S\;\pi-I$ feeding than the other feeds. In case of the feed of PSB and baker's yeast docosahexaenoic acid $(C_{22:6\omega3})$ did not show. In amino acid analysis, the rotifer fed on Erthrobacter sp, $S\;\pi-I$ showed more amino acid content comparing to those fed on other diets. Especially, arginine, isoleucine, histidine, lysine, methionine, phenylalanine, threonine, which are essential amino acid for fish growth, showed high contents. These results suggested that the aerobic photosynthetic bacterium, Erythrobacter sp. $S\;\pi-I$ would be a beneficial microbial teed for the cultivation of rotifer.

• Microbiology and Biotechnology Letters
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• 제8권3호
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• pp.155-166
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• 1980

Studies have teen undertaken to investigate the degree of microbial contamination in the subsidiary materials which have been known as an important source of microorganisms associated with spoilage of fish sausage and fish paste products. Twenty hinds of food ingredients including starch, spices and condiments, 59 samples in total collected from commercial fish sausage processing plants and supermarket in the period of July to October 1979, were examined for standard plate count, coliform and fecal coliform, mold and yeast, thermoduric microorganisms, aerobic sporeformers (mesophilic and thermophilic), anaerobic sporeformers (mesophilic and thermophilic) and sulfide spoilage anaerobes. The results obtained are summarized as follows. 1. Among the food ingredients examined, corn starch, black pepper, hot pepper, onion, garlic, ginger, beef extract and frank marked high bacterial contamination with general and sporeforming microorganisms. And bacterial content of marked samples were generally higher than that of the samples from plants. 2. The high standard plate count caused by high content of these bacteria like thermoduric, mesophilic or thermophilic sporeforming aerobes. 3. Bacterial content of food ingredients such as black pepper and beef extract being used in plants, and black pepper, hot pepper, onion and garlic from the market were exceeded the bacterial standards being enforced in Japan and U. S. A. 4. Average standard plate count was in the range of 10$^4$to 10$^{5}$ /g for black pepper, wheat flour, onion and garlic collected from plants, and 10$^{5}$ to 10$^{7}$ /g for black pepper, hot pepper, onion and garlic from market. No plate count was observed in pepper essence and coloring material. 5. Coliform organism was detected in starch, black pepper, hot pepper, onion, garlic, ginger and gluten that showed high standard plate but no fecal coliform in the samples except black pepper and hot pepper. 6. Average mold and yeast count was 140 to 460/g for corn starch, wheat flour and black pepper from plants, and 10$^3$/g for black pepper and hot pepper from market. No count was observed in the other ingredients. 7. Sulfide spoilage sporeforming anaerobes boiled for 5 min. at 10$0^{\circ}C$ and incubated at 55$^{\circ}C$ was not detected in all the samples examined.

• Journal of fish pathology
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• 제23권2호
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• pp.199-209
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• 2010

This study was aimed to investigate the effects in different immersion doses of garlic, Allium sativum, juice to modulate on the nonspecific immune responses of the olive flounder, Paralichthys olivaceus, artificially prechallenged with Streptococcus iniae BS10 and Edwardsiella tarda KE-1, respectively. The nonspecific immune responses of the tested fish were assessed in term of skin mucus lysozyme activity, the change of bacterial cell counts in organs, the number of lymphocytes and neutrophils in blood, and SOD activity. Almost groups of the prechallenged with either S. iniae BS10 or E. tarda KE-1 fish which had been immersed in garlic juice showed the enhanced skin mucus lysozyme activity, the number of lymphocytes and neutrophils in blood, and SOD activity in the kidney but the decreased the number of bacterial cell in surveyed organs. RPS in the group immersed in 0.25 g/L of garlic juice was much higher than in other immersed test groups. These results suggested that the garlic juice immersion can be effective on enhancement of the nonspecific immune responses and the protective ability of olive flounder to the artificial challenge with S. iniae BS10 and E. tarda KE-1.

• Journal of fish pathology
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• 제19권3호
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• pp.267-276
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• 2006

Seventy-five streptococci were isolated from diseased olive flounder, Paralichthys olivaceus in Jeju. Their drug susceptibility and transferable multiple drug resistance were characterized. All isolates were resistant to flumequine (AR) and oxolinic acid (OA) and 26 isolates (34.7%) showed 4～6 multiple resistance of ampicillin (ABPC), AR, doxycycline (DOXY), erythromycin (EM), norfloxacin(NOR), OA and oxytetracycline (OTC) in various combinations. pST9 of a transferable R plasmid was detected from a multiple drug resistance strain, Streptococcus sp., ST9 originated from diseased flounder in Jeju, previously. We performed DNA hybridization to know the distribution of plasmid with the same DNA structure as pST9 in streptococci. Thirteen out of 60 isolates analyzed were positive in colony DNA hybridization and the part of bacteria isolated from raw meal was also hybridized with pST9. It suggested that raw meal is one of the origin of the resistance plasmid and R plasmid with DNA structure differing from pST9 is also involving in multiple drug resistance of the streptococci. In conjugation experiment, we found transferable R plasmid carrying OTC, DOXY and/or EM resistance determinant in the 13 resistance strains. all of the streptococci carrying the transferable R plasmid were similar in RAPD patterns. However, pST -type R plasmid was rare in S. iniae most frequently appearing in flounder farm.

• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.994-1001
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• 2001

An easier way of understanding the BNR system was proposed from the study on substrate, nutrient removal tendency, microbial community and its metabolic function by applying the municipal settled sewage. During the anaerobic period, the phosphorus release rate per VFACOD we varied depending on the phosphorus content in the sludge. When the phosphorus content in the sludge was $6\%$ VSS, according to influent VFACOD, the phosphorus release rate and PHA production were $0.35 gPO_4P/gVFACOD$ and 1.0 gPHA/gVFACOD, respectively. The $NO_3N$ requirement for the phosphorus uptake as an electron acceptor was about $0.5 gNO_3N/gPO_4P_{uptake}$ based on the proposed equation with PHA, biomass, production, and the concentration of phosphorus release/uptake. Bacterial-community analysis of the sludge, as determined by FISH and 16SrDNA characterization FISH, revealed that the beta-subclass proteobacteria were the most abundant group ($27.9\%$ of the proteobacteria-specific probe EUB338), and it was likely that representative of the beta-subclass played key roles in activated sludge. The next dominant group found was the gamma-protebacteria ($15.4\%$ of probe EUB338). 16S rDNA clone library analysis showed that the members of${\beta}$- and ${\gamma}$-proteobacteria were also the most abundant groups, and $21.5\%$ (PN2 and PN4) and $15.4\%$ (PN1 and PN5) of total clones were the genera of denitrifying bacteria and PAO, respectively. Prediction of the microbial community composition was made with phosphorus content (Pv, $\%$ P/VSS) in wasted sludge and profiles of COD, PHA, $PO_4P,\;and\;NO_3N$ in an anaerobic-anoxic SBR unit. Generally, the predicted microbial composition based upon metabolic function, i.e., as measured by stoichiometry, is fairly similar to that measure by the unculturable dependent method. In this study, a proposal was made on he microbial community composition that was more easily approached to analyze the reactor behavior.

• Korean Journal of Fisheries and Aquatic Sciences
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• 제31권6호
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• pp.914-919
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• 1998

This study was conducted to evaluate the purification efficacy by filter and ultra violet (UV) irradiation against inlet and outlet seawater of the land based fish farm. Purification efficacies for inlet seawater (50$\ell$/min) has been examined with filter of 60$\mu$m pore size and UV irradiation at an average dose 0.5 mWS/$cm^2$ for 5 months. For outlet seawater filter of 90 $\mu$m pore size only was used. Temperature, salinity, pH, $NO_3-N,\;NO_2-N,\;NH_4-N,\;PO_4-P$, DO and COD of inlet and outlet seawater in the system were not influenced. However, the removing rate of suspended solid and turbidity of inlet seawater were appeared to be $48.7\~65.6\%$ (average $51.9\%$) and $33/3\~42/5\%$ (average $34.8\%$) after passing through screen filter respectively. Also, germicidal efficiency to the total bacteris and Vibrio species were $16.7\~20.2\%$ (average $19.2\%$) $20.0\~21.9\%$ (average $20.9\%$) respectively after passing through UV irradiation. After passing through drum filter of 90$\mu$m pore size, suspended solid and turbidity of outlet seawater were appeared to be $42.7\~52.6\%$ (average $46.9\%$), $27.7\~29.9\%$ (average $28.3\%$), respectively.