• Journal of Life Science
• /
• v.20 no.2
• /
• pp.190-196
• /
• 2010

Nowadays many people use antibiotics to protect processed foods from many pathogenic bacteria. The abuse of antibiotics, however, can run the risk of creating resistant forms of bacterium. Our study focus is on making new substances that can not only replace antibiotics but also be friendly to the environment. In our experiments, we used fermented citrus fruit, soil microbes and coenzyme Q10 as probiotics and prebiotics. Chickens in the experimental group were fed these substances via oral route while those in the control group were not. After specific time periods, blood and feces samples were collected to test for Salmonella spp.. It is interesting that fermented citrus fruit was the most effective in suppressing this bacterium. Furthermore, dissection of the experiment group chickens shows that their livers did not change to a yellow color, in contrast to the control group. The results confirmed our proposal that the chickens fed with these materials can be protected from infection by Salmonella and other pathogens. These probiotics and prebiotics are highly practical because they are natural substances that can be easily recycled in the environment. It can also be used as an animal feed ingredient because of its safety.

• Journal of fish pathology
• /
• v.27 no.1
• /
• pp.17-24
• /
• 2014

To study the possible use of probiotics in fish farming, The in vitro and in vivo antibacterial effects of Pseudomonas aeruginosa MB I-3 (MB I-3) against the fish pathogenic bacterium Listonella anguillarum were evaluated. The inhibitory effects of MB I-3 against vibrios were investigated by the double layer method and the co-culture. The results showed that MB I-3 inhibited the growth of pathogenic vibrios including Listonella anguillarum, Vibrio alginolyticus, Vibrio cholerae, Vibrio fluvialis, Vibrio furnissii, Vibrio harveyi, Vibrio parahaemolyticus and Vibrio vulnificus. Extracellular substances obtained from the cultural supernatant of MB I-3 by ethyl acetate extraction showed inhibitory effects on L. anguillarum. The antibacterial substance of MB I-3 was evaluated to destroy the cell membrane of L. anguillarum in electron micrographs. The probiotic effects of MB I-3 was tested by exposing olive flounder (Paralichthys olivaceus) fry to L. anguillarum with or without MB I-3. The cumulative mortality of olive flounder fry infected with L. anguillarum was 24% in the group with MB I-3, while it was 46% in the control group without MB I-3. These results indicate that MB I-3 has potential applications as a probiotic for the control of fish pathogenic vibrios in fish rearing system.

• Microbiology and Biotechnology Letters
• /
• v.38 no.2
• /
• pp.144-150
• /
• 2010

This study was aimed to screen bacteria of high alginate-degrading activity, to select the nitrogen source and concentration of NaCl and sodium alginate for the production of alginate-degrading enzyme, and to determine reaction conditions of enzyme. A novel alginate-degrading bacterium was isolated from abalone (Haliotis discus hannai) and named Methylobacterium sp. HJM27 by 16S rDNA sequence analysis. The optimum culture conditions for the production of alginate-degrading enzyme were 1.0% sodium alginate, 0.5% peptone, 0.3% yeast extract, 1.5% NaCl, $25^{\circ}C$ and 48 hours incubation time. The raw enzyme showed the highest activity at $25^{\circ}C$ and pH 9, and produced 1.217 g - reducing sugar per liter in 0.8% (w/v) sodium alginate for 30 minutes. Methylobacterium sp. HJM27 and its alginate-degrading enzyme would be useful for the production of bioenergy and biofunctional oligosaccharides from seaweed.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.42 no.3
• /
• pp.434-440
• /
• 2013

This study was conducted to screen the characteristics and alginate-degrading activity of marine bacterium isolated from brown seaweed (Sargassum thunbergii). The results of 16S ribosomal RNA sequence analysis the strain the genus Vibrio and the strain was subsequently named Vibrio sp. PKA 1003. The optimum culture conditions for the growth of Vibrio sp. PKA 1003 were at pH 7, 3% NaCl, $25^{\circ}C$, and 6% alginic acid, with a 48-hour incubation time. A crude enzyme preparation from Vibrio sp. PKA 1003, showed its highest levels of alginate-degradation activity when cultured at pH 9, $30^{\circ}C$, and 6% alginic acid, with a 63-hour incubation time. Thin layer chromatography analyses confirmed that the crude enzyme released monomers or oligomers from sodium alginate, and results from trypsin treatment showed that the alginate degrading activity depends on this enzyme produced by Vibrio sp. PKA 1003. These results suggest that Vibrio sp. PKA 1003 and its alginate-degrading crude enzyme is useful for the production of alginate oligosaccharides.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.31 no.5
• /
• pp.767-773
• /
• 1998

In this study, we have investigated the distributions and killing effects of marine bacteria that tend to kill the red tide microalgae, C. polykikoides in the area of Masan bay from June to October, 1996. To summarize, C. polykikoides killing bacteria were detected at $10^2$ to $10^3$ cells/ml of seawater samples during the survey period, and the bloom was observed in September by containing $4.8\times10^3$cells/ml. It appears however that the number of these bacteria is decreased ($2.0\times10^2$cells/ml) in October, A total of 110 strains were isolated from seawater samples and seawater filtrate (pore size, 0.8 $\mu$m)-containing mixed culture of C. polykikoides in which the mixed culture was grown in f/2 medium. As results we have successfully isolated Micrococcus sp. LG-1 which decreased to less than 10cells/ml within 6days and 5days sfter inoculation of Micrococcus sp. LG-1 into the la9 and logarithmic growth phases of C. polykrikoides respectively. Therefore, it appears that inoculation of Micrococcus sp. LG-1 against the logarithmic C. polykrikoides is more effective than the lag growth phase, (n addition, the killing effects were increased in accordance with bacterial cell densities inoculated in a dose dependent manner. Especially, the filtrate of kitling bacterium culture (nore size, 0.2 $\mu$m) revealed a dramatic effect in which C. polykrikoides were decreased to less than 10 cells/mf of culture within 1 hr, 1,5 hrs, 1,5 hrs, 3.5 hrs. and 5,5 hrs after inoculations of the culture filtrate with concentration of $30\%,\;20\%,\;10\%,\;5\%$ and $2.5\%$, respectively. Moreover Micrococcus sp. LG-1 showed a selective specificity against C. polykrikoides and any other killing effects of Micrococcus sp. LG-1 were not observed against Alexandrium tamarense, Prorocentrum micans, Scrippsiella trochoidea. ana Gymnodinium sanguineum.

• Korean Journal of Microbiology
• /
• v.54 no.1
• /
• pp.60-68
• /
• 2018

A cold-active and alkaline serine protease (Pro21717) was partially purified from the Antarctic marine bacterium Pseudoalteromonas arctica PAMC 21717. On a zymogram gel containing skim milk, Pro21717 produced two distinct clear-zones of approximately 37 kDa (low intensity) and 74 kDa (high intensity). These were found to have identical N-terminal sequences, suggesting they arose from an identical precursor and that the 37 kDa protease might homodimerize to the more active 74 kDa form of the protein. Pro21717 displayed proteolytic activity at $0-40^{\circ}C$ (optimal temperature of $40^{\circ}C$) and maintained this activity at pH 5.0-10.0 (optimal pH of 9.0). Notably, relative activities of 30% and 45% were observed at $0^{\circ}C$ and $10^{\circ}C$, respectively, in comparison to the 100% activity observed at $40^{\circ}C$, and this enzyme showed a broad substrate range against synthetic peptides with a preference for proline in the cleavage reaction. Pro21717 activity was enhanced by $Cu^{2+}$ and remained stable in the presence of detergent surfactants (linear alkylbenzene sulfonate and sodium dodecyl sulfate) and other chemical components ($Na_2SO_4$ and metal ions, such as $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $K^+$, and $Na^{2+}$), which are often included in commercial detergent formulations. These data indicate that the psychrophilic Pro21717 has properties comparable to the well-characterized mesophilic subtilisin Carlsberg, which is commercially produced by Novozymes as the trademark Alcalase. Thus it has the potential to be used as a new additive enzyme in laundry detergents that must work well in cold tap water below $15^{\circ}C$.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.33 no.4
• /
• pp.339-347
• /
• 2000

An algicidal bacterium, Micrococcus sp. LG-5 against the harmful dinoflagellate, Cochlodinium polykrikoides was isolated. The optimal conditions for the highest algicidal activity of bacterial culture filtrate showed in the range of $20{\~}30^{\circ}C$, at pH 7.0 and $3.0{\%}$ of NaCl concentration. In addition, $IC_(50)(mean of 50{\%} inhibitory concentration)$ of the culture filtrate against C. polykrikoides after incubation of 5 days was $0.482{\%}$. To investigate heat and pH stability of the culture filtrate of Micrococcus sp. LG-5, the culture filtrate ($pore size, 0.1 {\mu}m$) was heated to $121^{\circ}C for 15 min$ and adjusted pH from 2.0 to 10.0. There were no significant changes in algicidal activity by heat treatment and the pH change between pH from 5.0 to 10.0. The algicidal substances produced from Micrococcus sp. LG-5 were mainly detected in the fraction of $10,000{\~}1,000$ MWCO (molecular weight cut-off). The culture filtrate of Micrococous sp. LG-5 showed antimicrobial activity against Enterococcus faecalis, Escheiichia coli, Uebsiella pneunioniae and Vibrio altinolyticus, but did not show against Pseudomonas aeminosa, P. Buorescens, Salmonella typhi, Staphylococcus aureus, V. cholerae and V parahaemolyicus. The culture filtrate of Micrococcus sp. LG-5 was examined against 16 phytoplankton species and showed the algicidal activity against Ajexandzium tuarense, Eutreptiella Drnnastin, Gymnodinium catenatum, G. mikimotoi, G. sanguineum, eyodinium impuaicum, Heterocapsa triquetra, Heterosipa akashiwo, Prorocentrum micans and Pyraminonas sp.. However no algicidal effects of Micrococcus sp. LG-5 were observed against Chlamydomonas sp., Cylindrotheoa closterium, P. mininum, P. triestimum, Pseudonieschia sp. and Sczipuiella trochoidea. On the other hand, algicidal activity on the tested marinelivefood was not detected except for Isochrysis galbana. In addition, physiological responses of cultured olive flounder (Paralichthys oliraceus) exposed to $1 and 10{\%}$ of the culture filtrate of Micrococcus sp. LG-5 were measured. There were no clear changes in AST, GGT, creatinine, urea, total cholesterol, total protein, albumine, $Mg^(+2), Ca^(+2), Na^+, K^+, and Cl^-$. These results indicate that olive flounders were not affected when they were exposed to the culture filtrate of Micrococcus sp. LG-5.