Kim, Min-Sun;Park, So-Hyun;Kim, Dong-Hwi;Heo, Moon-Soo
Journal of Life Science
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v.24
no.8
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pp.880-886
/
2014
In this study, we isolate and identify bacteria from seawater collected from Jeju coast, to evaluate the antimicrobial activity against the fish pathogenic bacteria. 14 bacterial strains were isolated and identified using physiological, biochemical and molecular tools. Antibacterial activity of all the 14 isolates were screened against four major fish pathogens namely, two Gram-positive: Streptococcus iniae, Streptococcus parauberis and two Gram-negative: Vibrio anguillarum, Edwardsiella tarda. Results revealed that among the 14 isolates, MK-11 was found to have antibacterial activity against S. iniae, S. parauberis, V. anguillarum Particularly, S. iniae was susceptibility with the MIC value of $250{\mu}g/ml$. The biochemical and physio-chemical results reveal that MK-11 had the sugar-alcohol disassemble ability of the D-sorbitol and D-mannitol. Also the utilization of the yeast extract, sorbitol and di-potassium phosphate were noted to be high. The optimum culture condition such as pH and temperature was recorded as pH 6.0, $25^{\circ}C$ and along with 1% NaCl which differs from the previous reports particularly in nutrient resolutions. As results of the analysis of 16S rDNA sequences, MK-11 show the high similarity with Paenibacillus polymyxa, P. jamilae, P. brasilensis 99.78%, 99.43%, 99.39%, repectively. Hence, in the present study, the isolated Paemibacillus sp. MK-11 from Jeju seawater possesses the antibacterial activity against fish pathogens and it could be used as a new antibiotic agents against the gram positive fish pathogens.
This study aimed to compare the disinfection efficiencies of the ultraviolet and plasma systems, the two systems designed and commercialized to disinfect water in aquaculture, by putting each in a 100 ℓ water tank and concentrating 1.0 ℓ of treated water to check the changes in the number of bacteria in the samples. Each system was operated for 6 hours to culture the typical seawater bacteria in the Marine agar, Thiosulfate citrate bile salts sucrose agar and Salmonella Shigella agar media, respectively, to check the number of bacteria in the media, and the changes in the number of Edwardsiella piscicida in the treated water were checked after the artificial inoculation of E. piscicida in the disinfected seawater. As a result, the two disinfection systems showed the almost similar levels of bacterial reduction efficiency between 99.5% and 99.9%. However, the result of this study showed that, with 100 ℓ of water treated for the same length of time using the two systems, the plasma system turned out to disinfect bacteria in a shorter period of time than the UV system. However, as the changes in the number of bacteria were checked for a short length of time (6 hours) in this study, it was judged that, considering the actual aquaculture environment in which the quality of water significantly changes with feed residues, excretions and coastal contamination, etc., and a lot of biofilms and organic matter exist, the plasma system would be more efficient than the UV system as the former is capable of continuously maintaining a certain level of efficiency than the latter that is limited in terms of efficiency depending on the level of turbidity and the existence of organic matter.
Considerable damages of the cultivated Porphyra by Porphyra diseases were reported from the Porphyra culture bed along the coast of Yongwon-ri, Changwon, Kyungnam during the years of 1969 and 1970. The present study deals with the effects of fertilizer plant effluents on the Porphyra diseases, and the results are summarized as follows : 1. By the result of current observation, the polluted water of Haeng-am Bay which has an inflow of pollutants from the fertilizer plant was affecting the bed with tides. The results of pollution studies in Chinhae Bay and adjacent waters conducted by Won and Park(1971) also show that Chinhae Bay and adjacent waters are contaminated with the plant effluents. It seems that the effect increases due to the wind drift current when north-westerly or westerly winds prevail. Accordingly, effects of the Porphyra diseases in the culture bed seem to originate from the pollutants, since there are more damages when the north-westerly or westerly winds prevail, and also spring tide develops. 2. As compared to the photosynthetic activity of the Porphyra suborbiculata in uncontaminated seawater, it decreases up to $4\%$ in 200pmm, $20\%$ in 300ppm and $43\%$ in 1,000ppm of contaminated seawater which contains dilluted pollutants from the fertilizer plant. The results of the measurements using the water collected in the polluted area of Chinhae Bay and adjacent waters revealed that the photosynthesis was depressed about 21 to $34\%$ near the plant, and in the area of the Porphyra beds, $15\%$ in the portion where tide is weak and $5\%$ where the tide is strong, in comparison with the area of unpolluted water. 3. Although the present results do not indicate the exact level of harmful pollutants, it is evident that the pollutants of the plant effluents interfere photosynthetic activity of the Porphyra, even in the water containing pollutants as low as 200 ppm.
SHIN, AYOUNG;KIM, DONGSUNG;KANG, TEAWOOK;OH, JE HYEOK
The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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v.25
no.2
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pp.26-41
/
2020
In order to culture a life for the physiological and ecological research of the meiofauna, this study aimed to identify the most ideal condition in which the meiofauna can be cultured within a laboratory by setting various environmental conditions. The sediment deposits and seawater were collected from the intertidal zone in Mallipo of the west coast. A aquarium in which the internal environment can be controlled by constantly maintaining the temperature and humidity was fabricated and the culture experiments of the collected meiofauna were conducted together with the sea water and sediment deposits collected. The experiment 1 was conducted after establishing the similar environment as the collecting location. Under the same condition as the experiment 1, the experiment 2 verified a difference between when live foods were supplied and were not. In the experiment 3, the changes in the meiofauna colony were checked according to with or without light and live foods. In the results of culturing experiments, the habitat density and the number of appeared classification groups of the meiofauna colony were relatively higher both in the water tank with supplying the live foods and under the condition of having light in 12-hour cycle than those in the aquarium without live foods and under no light condition. In addition, the habitat density of meiofauna cultured within a laboratory exhibited relatively higher value than that under the natural state.
An agar-degrading marine bacterium, strain AS-1 was isolated from the seawater. The strain AS-1 was identified as Sphingomonas paucimobilis (90% probability) by VITEK. The optimum medium for agarase activity of the isolated strain was determined to be marine medium, marine broth 2216 containing 0.1% agar as carbon source. An extracellular agarase was purified 104-fold from the culture supernatant by ammonium sulfate precipitation, ion exchange chromatography and gel filtration methods. The molecular weight of the purified enzyme was estimated to be 80 kDa by SDS-PAGE. The optimum pH and temperature for activity were 7.0 and $40^{\circ}C$, respectively. Antioxidative activity of the strain AS- was 72% in the supernatant cultured for 12 h. The culture supernatant of the strain AS-1 showed antibacterial activity against bacteria causing putrefaction and food poisoning such as Escherichia coli, Staphylococcus aureus and Proteus vulgaris. However, the cell growth of the lactic aicd forming strain, Lactobacillus plantarium was promoted by the treatment of 10% culture supernatant of an agar-degrading strain.
An agar-degrading marine bacterium, strain AS-3 was isolated from the seawater. The strain AS-3 was identified as Algibacter lectus AS-3 by 16S rDNA sequence. The optimum medium for agarase activity of the isolated strain was determined to be marine medium, marine broth 2216 containing 0.1% agar as carbon source. An extracellular agarase was purified 6.9-fold from the culture supernatant by ammonium sulfate precipitation, ion exchange chromatography and gel filtration methods. The optimum pH and temperature for this enzyme were 7.0 and $40-50^{\circ}C$, respectively. Antioxidative activity of the strain AS-3 was 62.4% in the supernatant cultured for 12 h.
A $\beta$-agarase gene, agaB34, was functionally cloned from the genomic DNA of a marine bacterium, Agarivorans albus YKW-34. The open reading frame of agaB34 consisted of 1,362 bp encoding 453 amino acids. The deduced amino acid sequence, consisting of a typical N-terminal signal peptide followed by a catalytic domain of glycoside hydrolase family 16 (GH-16) and a carbohydrate-binding module (CBM), showed 37-86% identity to those of agarases belonging to family GH-16. The recombinant enzyme (rAgaB34) with a molecular mass of 49 kDa was produced extracellularly using Escherichia coli $DH5{\alpha}$ as a host. The purified rAgaB34 was a $\beta$-agarase yielding neoagarotetraose (NA4) as the main product. It acted on neoagarohexaose to produce NA4 and neoagarobiose, but it could not further degrade NA4. The maximal activity of rAgaB34 was observed at $30^{\circ}C$ and pH 7.0. It was stable over pH 5.0-9.0 and at temperatures up to $50^{\circ}C$. Its specific activity and $k_{cat}/K_m$ value for agarose were 242 U/mg and $1.7{\times}10^6/sM$, respectively. The activity of rAgaB34 was not affected by metal ions commonly existing in seawater. It was resistant to chelating reagents (EDTA, EGTA), reducing reagents (DTT, $\beta$-mercaptoethanol), and denaturing reagents (SDS and urea). The E. coli cell harboring the pUC18-derived agarase expression vector was able to efficiently excrete agarase into the culture medium. Hence, this expression system might be used to express secretory proteins.
LEE Kyun Woo;PARK Heum Gi;LEE Sang-Min;HAN Hyon Sob;LIM Young Soo
Korean Journal of Fisheries and Aquatic Sciences
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v.37
no.1
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pp.7-12
/
2004
This study investigated the possibility of salinity acclimation of freshwater rotifers (Brachionus calyciflorus) as live food for sweetfish (Plecoglossus altivelis) larvae, and also examined the optimal salinity for the growth of sweetfish. Freshwater rotifers cultured in 0 and 4 PSU and seawater rotifers (B. rotundiformis) cultured in 33 PSU were supplied to the larvae with four kinds of enrichment material (condensed freshwater Chlorella, $\omega-yeast,$ baker's yeast, Super Selco) and larval growth at 4 PSU was examined. Growth of the freshwater rotifers positively increased from 0 PSU to 6 PSU, but decreased when over 8 PSU was reached. Growth and survival of the sweet fish larvae reared in 0 PSU were significantly lower than those reared in either 4 PSU or 33 PSU. This indicated that the freshwater rotifers (B. calyciflorus) could be used as live food for sweetfish larvae reared in 4 PSU. The body weight of sweetfish larvae fed on freshwater rotifers enriched with Super Selco was the highest at 0.163 mg, but there was no significant difference in survival and body length of the fish fed with the other enrichment materials. The content of n-3 HUFA of the sweetfish larvae fed on the freshwater rotifers enriched with Super Selco and the condensed freshwater Chlorella was higher than that enriched with $\omega-yeast$ and baker's yeast. These results indicated that B. calyciflorus cultured with the condensed freshwater Chlorella could be used for the sweetfish larvae without enrichment, and the most efficient enrichment material for B. calyciflorus is Super Selco.
Five urease-positive Vibrio parahaemolyticus strains were isolated from Kamak Bay in Yeosu in 2002 and 2003. V. parahaemolyticus YKB4 and YKB14 were isolated from seawater, YFB20 from black rockfish (Sebastes schlegeli), and YFO2l and YFO22 from olive flounder (Paralichthys olivaceus). The five urease-positive strains (YKB4, YKB14, YFB20, YFO21, and YFO22) did not show hemolysin and protease activity, while they did alter in color (to red) as the bacteria grew in the urea broth medium. All samples showed identical biochemical characteristics as a reference strain, V. parahaemolyticus KCTC2471, except in urease production. The five urease-positive strains showed urease activities at a mid stationary phase, and their activity was maximal in the late stationary phase of their culture supernatant. The addition of urea to the Luria-Bertani (LB) broth medium significantly affected the initial production of urease of V. parahaemolyticus isolates. Mortality by urease-positive V. parahaemolyticus YKB4, YKB14, YFO2l, and YFO22 was significantly high, being$60-80\%$, while YFB20 only reflected a rate of $20\%$. Protease-positive V. parahaemolyticus FM39 and FM50 showed a $40\%$ and $60\%$ mortality rate, respectively. However, hemolysin-positive V. parahaemolyticus had no mortality, like the non-pathogenic V. parahaemolyticus KCTC2471, while V. vulnificus resulted in a $40\%$ mortality rate. Injection with urease-positive V. parahaemolyticus strains showed mortality within 12 hrs in mice, and the strains could be isolated from the dead mice.
The effects of sparp changes of water temperature (WT) on the stree response and physiological change of the cultured olive flounder (Paralichthys olivaceus) were examined by manipulating WT (3 patterns) in a running seawater culture system. In the first group (Exp. I), the WT was decreased from 18$^{\circ}C$ to 11$^{\circ}C$ within 6 hours and increased back to the original WT quickly. WT was decreased from 2$0^{\circ}C$ to 11$^{\circ}C$ within 5 hours and main-tained at 11$^{\circ}C$ for 10 hours. and then increased to 2$0^{\circ}C$ in the second group (Exp. II). In the third group(Exp. III) WT was decreased to 11$^{\circ}C$ within 5 hours (type A) or 10 hrs. (type B). In Exp. I and III, the level of serum cortisol was increased from 2.5$\pm$0.3 ng/ml and 2.6$\pm$0.9 ng/ml to 13.6$\pm$3.0 ng/ml and 12.4$\pm$3.2ng/ml, respectively, with WT decrease. However, no consistent tendency in the change of serum glucose level was shown according to WT decrease. In Exp. III, the glucose level of fish in type A was decreased until 5 hours and increased at 7 hours, then decreased until 12 hours where as the glucose level in type B was decreased until 5 hours and stayed at the level of 15.7 mg/dl. The serum osmolality was reduced with WT decrease and the response of serum electrolytes in this experiment conflicted, and a tendency in total protein, AST and ALT was not found following WT decrease. In conclusion, olive flounder responded to the stress caused by WT decrease and acclimated to this condition when the lower temperature was maintained. But there was no stress response in the blood of olive flounder when WT was increased. On the other hand, the degree of stress response in olive flounder was various according to the range and gradient of WT change.
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