The biological characteristics of Bacillus sp.CPB-St as a probiotic strain to control fish streptococcosis was determined. Based on 16S rRNA sequencing, Bacillus sp.CPB-St was identified as Bacillus pumilus and named B. pumilus CPB-St (Abbreviated as CPB-St). Growth inhibitory activity of CPB-St against Streptococcus spp. was examined at three different incubation temperatures ($20^{\circ}C$, $25^{\circ}C$, and $30^{\circ}C$) and three culture media (NA, TSA, and BHIA) based on the diameter of inhibition zone. Its activity (inhibition zone of 11~29 mm) at $20^{\circ}C$ was higher than that (12~21 mm) at $30^{\circ}C$. Its activity (29 mm) in NA media was the same as that (29 mm) in TSA media. However, it was higher than that (22 mm) in BHIA media. The inhibitory activity of CPB-St against Streptococcus spp. was high at pH7. However, its activity was the same at salinity of 0.5% to 3%. CPB-St showed maximum growth after incubation at $25^{\circ}C$ for 48 h. To use CPB-St as probiotics, settlement studies in fish intestine and its efficacy through feeding are needed. CPB-St was highly resistant to gastric juice at pH4 and flounder's bile salt as well as deoxycholic acid at $300{\mu}g/ml$. CPB-St showed optimal viability in 1% NaCl. It showed similar growth in 0% to 7% NaCl. CPB-St could tolerate $-20^{\circ}C$ and $-70^{\circ}C$ for 45 min. There was no difference in the growth of the strain between room temperature and $4^{\circ}C$. Fish diet supplemented with CPB-St could be stored at low temperature without cell loss. Therefore, CPB-St might be used as probiotics to control streptococcosis of fish.
The cell density changes of Vibrio mimicus K-1 in sea water and arkshell feeding it were examined at various temperature. The strain was suspended in sterilized sea water and storaged at experimental temperature $(5,\;10,\;15,\;20,\;and\;28^{\circ}C)$). At intervals of up to 10 days, aliquots of each suspension were plated onto BHI agar. At 5 and $10^{\circ}C$, the plate counts of V. mimicus K-1 showed a rapid decline, which 3s known to be a reault of this bacterium's entering into the viable but non culturable state. At 20 and $28^{\circ}C$, however, V. mimicus K-1 are stable over the 10 days experimental periods. V. mimicus K-1 was fed to arkshell, which was subsequently stored at temperatures ranging from 5 to $20^{\circ}C$ for 10 days. The samples of arkshell were homogenized and plated at intervals to determine the cell density of V. mimicus K-1 and total aerobic population of bacteria present. At 5 and $10^{\circ}C$, the numbers of V. mimicus K-1 in sea water rapid decreased over the 10 days experimental periods. However, little change of V. mimicus K-1 density was observed in shellstock arkshell at 5 and $10^{\circ}C$. While, V. mimicus K-1 density was decreased more rapidly to level below limit of dectection in shucked arkshell at same temperature. Incubation at the higher temperature $(20^{\circ}C)$ resulted in large increase in total aerobic bacterial number of shellstock arkshell. These results suggest that even with proper storage, indigenous levels of V. mimicus may remain sufficiently high in shellstock arkshell to produce infection in compromise hosts.
Disposal of high amount of coal combustion by-products, such as fly ash and bottom ash, is of a great concern to the country, due to the huge treatment cost and land requirement. On the other hand, those coal-ash wastes are considered to have desirable characteristics that may improve physical, chemical, and biological properties of soils. Especially, compared with fly ash, bottom ash has a larger particle size, porous surface area, and usable amount of micronutrients. In the present study, we examined bottom as a soil amendment for mitigating $CO_2$ emission and enhancing carbon sequestration in soils fertilized with organic matter (hairy vetch, green barely, and oil cake fertilizer). Through laboratory incubation, $CO_2$ released from the soil was quantitatively and periodically monitored with an enforced-aeration and high-temperature respirometer. We observed that amendment of bottom ash led to a marked reduction in $CO_2$ emission rate and cumulative amount of $CO_2$ released, which was generally proportional to the amount of bottom ash applied. We also found that the temporal patterns of $CO_2$ emission and C sequestration effects were partially dependent on the relative of proportion labile carbon and C/N ratio of the organic matter. Our results strongly suggest that amendment of bottom ash has potential benefits for fixing labile carbon as more stable soil organic matter, unless the bottom ash contains toxic levels of heavy metals or other contaminants.
In this study, we constructed a biological system that exhibited thermotolerance, ethanol tolerance, and increased ethanol productivity using a random mutagenesis method. We attempted to isolate a thermotolerant mutant using proofreading-deficient DNA polymerase ${\delta}$ and ${\varepsilon}$ encoded by the pol3 and pol2 genes, respectively, in Saccharomyces cerevisiae. To obtain mutants that could grow at high temperatures ($38^{\circ}C$ and $40^{\circ}C$), random mutagenesis of AMY410 (pol2-4) and AMY126 (pol3-01) strains was induced. The parental strains (AMY410 and AMY126) grew poorly at temperatures higher than $38^{\circ}C$. By stepwise elevation of the incubation temperature, AMY410-Ht (heat tolerance) and AMY126-Ht strains that proliferated at $40^{\circ}C$ were obtained. These strains were further incubated in medium containing 6% and 8% ethanol and then AMY410-HEt (heat and ethanol tolerance) and AMY126-HEt strain with ethanol tolerance at an 8% ethanol concentration was obtained. The AMY126-HEt strain grew even at an ethanol concentration of 10%. Furthermore, following the addition of high concentrations of glucose (5% and 10%), an AMY126-HEt3 strain with increased ethanol productivity was isolated. This strain produced 24.7 g/l of ethanol (95% theoretical conversion yield) from 50 g/l of glucose. The findings demonstrate that a new biological system (yeast strain) showing various phenotypes can be easily and efficiently bred by random mutagenesis of a proofreading- deficient mutant.
Green tides, which was mainly caused by Ulva spp., have been increasing in severity and frequency globally, and have negatively affected on marine ecosystems. This study was conducted to investigate effects of various physical and chemical factors on the death of Ulva australis (ULAUS) and to consider a practical measures useful for alleviating Ulva bloom. Soaking of ULAUS thalli in pure water for 8 hr didn't induce a death, but incubation in 1.0-1.5% salinity for 7 d inhibited sporulation by about 70%. Desiccation gave rise to a serious damage when more than 40-50% of initial fresh weight was lost. ULAUS growth was sensitive to temperature and seriously inhibited from more than $30^{\circ}C$. At $35^{\circ}C$, $40^{\circ}C$, $45^{\circ}C$ and $50^{\circ}C$, treatment time required for 90-95% death of ULAUS thalli was 1 d, 10 min, 30 sec, and 1 sec, repectively. ULAUS growth was seriously inhibited at lower than pH 6.0 and completely dead at pH 4.0. Several compounds for ULAUS control was selected and the chemcals causing a rapid death were oxidants such as hydrogen peroxide and sodium percarbonate. Taken together, our results suggest that low salinities, dryness, pH, high temp. and compounds could be selected for Ulva bloom control, and high temperature and compounds seems to be useful for a development of practical control methods.
Attempts to search infection period, infection speed in the tissue of neck blast of rice plant, location of inoculum source and effects of several conditions about the leaf sheath of rice plants for neck blast incidence have been made. 1. The most infectious period for neck blast incidence was the booting stage just before heading date, and most of necks have been infected during the booting stage and on heading date. But $Indica{\times}Japonica$ hybrid varieties had shown always high possibility for infection after booting stage. 2. Incubation period for neck blast of rice plants under natural conditions had rather a long period ranging from 10 to 22 days. Under artificial inoculation condition incubation period in the young panicle was shorter than in the old panicle. Panicles that emerged from the sheath of flag leaf had long incubation period, with a low infection rate and they also shown slow infection speed in the tissue. 3. Considering the incubation period of neck blast of rice plant, we assumed that the most effective application periods of chemicals are 5-10 days for immediate effective chemicals and 10-15 days for slow effective chemicals before heading. 4. Infiltration of conidia into the leaf sheath of rice plant carried out by saturation effect with water through the suture of the upper three leaves. The number of conidia observed in the leaf sheath during the booting stage were higher than those in the leaf sheath during other stages. Ligule had protected to infiltrate of conidia into the leaf sheath. 5. When conidia were infiltrated into the leaf sheath, the highest number of attached conidia was observed on the panicle base and panicle axis with hairs and degenerated panicle, which seemed to promote the infection of neck blast. 6. The lowest spore concentration for neck blast incidence was variable with rice varietal groups. $Indica{\times}Japonica$ hybrid varieties were infected easily compared to the Japonica type varieties, especially. The number of spores for neck blast incidence in $Indica{\times}Japonica$ hybrid varieties was less than 100 and disease index was higher also in $Indica{\times}Japonica$ hybrid than in Japonica type varieties. 7. Nitrogen content and silicate content were related with blast incidence in necks of rice plants in the different growing stage changed during growing period. Nitrogen content increased from booting stage to heading date and then decreased gradually as time passes. Silicate content increased from booting stage after heading with time. Change of these content promoted to increase neck blast infection. 8. Conidia moved to rice plant by ascending and desending dispersal and then attached on the rice plant. Conidia transfered horizontally was found very negligible. So we presumed that infection rate of neck blast was very low after emergence of panicle base from the leaf sheath. Also ascending air current by temperature difference between upper and lower side of rice plant seemed to increase the liberation of spores. 9. Conidial number of the blast fungus collected just before and after heading date was closely related with neck blast incidence. Lesions on three leaves from the top were closely related with neck blast incidence, because they had high potential for conidia formation of rice blast fungus and they were direct inoculum sources for neck blast. 10. The condition inside the leaf sheath was very favorable for the incidence of neck blast and the neck blast incidence in the leaf sheath increased as the level of fertilizer applied increased. Therefore, the infection rate of neck blast on the all panicle parts such as panicle base, panicle branches, spikelets, nodes, and internodes inside the leaf sheath didn't show differences due to varietal resistance or fertilizers applied. 11. Except for others among dominant species of fungi in the leaf sheath, only Gerlachia oryzae appeared to promote incidence of neck blast. It was assumed that days for heading of varieties were related with neck blast incidence.
This study was conducted to investigate the physiological properties of various parts of skate body after fermentation by measuring compositional properties including pH and $NH_4^+$. Other functional properties, such as antibacterial activities, antioxidative activities and anticancer activities were measured. Effects of fermentation temperature (4, 10, $20^{\circ}C$) did not affect compositional properties of fermented skate. The pH of fermented skate extract at 4$^{\circ}C$ did not increase as much as that at 10 and $20^{\circ}C$, Particularly at $10^{\circ}C$, the pH increased rapidly after Day 1 and remained unchanged until another increase at Day 5. At 2$0^{\circ}C$, the pH increased rather rapidly at early stage of fermentation and reached 8.9 at Day 4. The pattern of $NH_4^+$ concentration of fermented skate extract was similar to that of pH. Particularly at $4^{\circ}C$ fermentation, $NH_4^+$ concentration was not affected by fermentation time. The concentration of $NH_4^+$. reached approximately $10.2\mug/mL$at $10^{\circ}C$ for Day 5 and $20^{\circ}C$ for Day 4-5, indicating the early stage of fermentation. According to physiological activities of hot water extracts of skate fillet and viscera as affected by fermentation time, antibacterial activity of 2% viscera extract concentration was 43.3% at Day 8, while there was no antibacterial activity from fillet extract. As for the antioxidative activity, fillet extract and viscera extract both at 2% concentration at Day 0 showed 61.2% and 54.4%, respectively. Anticancer activities were highest (52.7% for fillet extract and 58.3% for viscera extract) at $1,000 \mug/mL$ concentration at Day 8. Antibacterial activities and anticancer effects were relatively high as fermentation was progressed, while antioxidative activities were highest before fermentation started. As for the physiological activities of hot water extract from brain and cartilage, antibacterial activities were observed at 41.0% when 2% brain extract was added at 4 hours of incubation, while 35.8% with 2% cartilage extract at 14 hours of incubation. Antioxidative activities were not found in brain extract, but cartilage extract at 2% showed 25.0% of antioxidative activity, which was lower than fillet and viscera extract.
Cells of Acinetobacter sp. strain JC1 DSM 3803, an aerobic monoxide-oxidizing bacterium, growing on glucose exhibited high catalase activity at the mid-exponential growth phase. The enzyme activity decreased gradually after then until the early stationary phase, increased again at the mid-stationary phase, and then decreased again thereafter. Cells growing on glucose was found to contain three kinds of catalses. Cat1, Cat2 and Cat3. The activities of Cat1 and Cat3 did change significantly during growth, but that of Cat2 exhibited significant variation. Cat3 was found to present only in cells growing on glucose, but not in cells growing on carbon monoxide of methanol. The activities of call and Cat3 in cell-free extracts were stable upon treatment with ethanol and chloroform, but decreased to some extent when the enzymewere treated with 2mM $H_2O_2$ and/or 3-amino-1,2,4-triazole (AT). Cat2 was found to be extremely sensitive to the ethanol-chloroform and $H_2O_2$ treatments, but was insensitive to the AT treatment. Cat1 exhibited enzyme activity after incubation for 1 min at 80$^{\circ}C$. Cat2 and Cat3 did not show enzyme activity after incubation for 1 min at 60$^{\circ}C$ and 70$^{\circ}C$, respectively. Cat2 was found to have peroxidase activity. Cat3 was purified to homogenity in seven steps. The molecular weight of the native enzyme was estimated to be 150,000. Sodium dodecyl sulfate-gel electrophoresis revealed two identical subunits of molecular weight 65,000. The enzyme was found to show two $K_m$ values of 39 mM and 58mM. The optimal pH for the enzyme activity was 7.0, but the activities at pH 6.0, 8.0, and 9.0, were found to be comparable to that at the optimal pH. The optimal temperature for the enzyme activity was found to be 40$^{\circ}C$. The enzyme also exhibited strong activity at 20$^{\circ}C$, 30$^{\circ}C$, and 50$^{\circ}C$. The purified enzyme was not affected by the ethanol-chloroform treatment. The enzyme, howerver, showed less than 10% of the original activity when it was treated with 12 mN AT, 0.1 mM $NaN_3$ of 1mM KCN.
'Gonji-2ho' a new variety of oyster mushroom, fitting for the bag culture, was bred by mating between monokaryons isolated from GMPO35338 and Jangpug. In the major characteristics of fruit body, the pilei were thick and dark-gray and the stipes were thick and long with softness. It was great in elasticity and cohesivness of tissue as compared to Suhan-1ho. The optimum temperature for the mycelial growth was around $26{\sim}29^{\circ}C$ and for the pinheading and growth of fruit body was around $14{\sim}18^{\circ}C$. In the bag culture, it was required around 20 days at incubation period and 5 days at primordia formation. The fruit body was grown vital and uniform. The yields were 323.3g/kg bag. This variety has high yielding capacity, cultivation stability and the resistance to the bacterial brown blotch disease.
The rationale for the use of fungi in treating waste streams from food processing plants I~as been that of incorporating the dissolved and suspending nutrients into a macroscopic organism which can be filtered out readily. In order for a process using fungi to meet these objectives we examined a strain of fungi, Aspergillus fumigatus, which grew well on a variety of polysaccharide-containing materials and showed both efficient BOD removal and high quality protein recovery. In this experiment the fungal choice was based on the laboratory screening studies where the criteria used was BOD and COD reduction, growth response, mycelial yield, and the ability to compete with the natural flora. In the fermentation system used far the continuous culture of Aspergillus fumigatus the best combination of operating variables, inoculum ratio, temperature, initial pH, fermentation time and agitation rate was 5%(v/v), $35{\sim}40^{circ}C,\;pH\;4.5{\sim}5.0$, 2days and 150rpm, respectively. The fungus reduced BOD and COD to 94.0 and 90.4%, respectively and 3.15g of dry mycelium per liter of alcohol waste was harvested during 48hr of incubation time. The protein efficiency ratios for the control diet and the experimental diet containing the fungal protein were $3.42{\pm}0.15$ and $3.40{\pm}0.43$, respectively.
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