Summer patch is the most serious disease at turfgrass field or golf course established with Kentucky bluegrass during high temperature season in Korea. Nevertheless, chemicals for the summer patch control are not yet registered in Korea. We isolated the pathogens from the turfgrass showing typical summer patch symptoms and identified as Magnaporthiopsis poae by using the internal transcribed spacer ITS1 and ITS4 sequences of rDNA. The inhibition rates of the pathogen were investigated for 10 fungicides. As results, the pathogen growth was suppressed when chemicals concentration increased and negatively correlated with incubation period with the chemicals. In triazole group, all chemicals (metconazole, myclobutanil, propiconazole and tebuconazole) treated showed the inhibition rates by 100%. Thiophanate-methyl showed the next highest inhibition effect against a summer patch pathogen. In strobilurin group, pyraclostrobin was the highest suppression effect compared with azoxystrobin and trifloxystrobin. Inhibition effect of fludioxonil and fluxapyroxad on pathogen was similar to the trifloxystrobin. Based on the results, triazole and carboxamide groups are strongly recommended due to the highest inhibition effect on the summer patch pathogen, Magnaporthiopsis poae.
To determine sex of bovine embryos using hamster histocompatibility Y(H-Y) antibodies, bovine compact morulae were incubated for 6 hours in TCM199 supplemented with 10% hamster H-Y antiserum and the embryos with developmental arrest were diagnosed as male embryos, while the embryos showing development during the incubation as female embryos. This presumptive embryo sexing was confirmed by polymerase chain reaction(PCR)method. 1. In the result of hamster sperm cytotoxicity test to measure H-Y antibody titer, the rate of dead sperm was considerably lower in H-Y antiserum absorbed with hamster male splenocytes than in H-Y antiserum absorbed with hamster female splenocytes or H-Y antiserum unabsorbed with splenocytes(p<0.01). 2. The rate of oocytes fertilized in vitro and the rate of blastocysts of the fertilized oocytes were 58.5% and 32.4%, respectively. The rate of blastocysts on day 8 was 15.9%, denoting the highest rate during whole culture period posterior to in vitro fertilization (IVF). 3. The bovine 16 cell and compact morulae embryos incubated in the medium supplemented with hamster H-Y antibodies showed 37.1% and 48.9% of developmental arrest which were diagnosed as male, respectively, and rates of redeveloped embryos from the arrested were 24.1% in 16 cell and 44.3% in compact morulae embryos, respectively, denoting higher rate of sex determination and rate of redevelopment in compact morulae than 16 cell embryos. 4. Bovine compact morulae of Korean cattle and Holstein were treated with hamster H-Y antibodies for sex determination and the rates of developmental arrest(diagnosed as male) were 48.4% for Korean cattle and 47.9% for Holstein, respectively. The rates of redeveloped embryos to blastocyst after treatment were 42.6% for Korean cattle and 41.8% for Holstein, respectively, showing no significant differences of sex determination and redevelopment between both breed. 5. The sex determination of bovine embryos(Korean cattle and Holstein) using hamster H-Y antibodies was diagnosed by PCR for confirmation, denoting the rates of 86.1% for Korean cattle and 85.9% for Holstein male embryos, respectively, and the rates of 91.9% for Korean cattle and 90.1% for Holstein female embryos, respectively, with no significant differences of sex determination between both breed. These results indicated that hamster H-Y antibodies can be usable for sex determination of bovine embryos of Korean cattle and Holstein, the viability of bovine embryos was sustained while being cultured in the medium supplemented with hamster H-Y antibodies of appropriate titer and sex determination of bovine embryos by PCR can be feasible for confirmation.
Bacteria in the viable but nonculturable (VBNC) state fail to produce colonies on routine bacteriological media, but are still alive in the state of very low metabolic activity. The aim of the present study was to induce the VBNC state of the Edwardsiella tarda using sea water microcosm under starvation conditions at $10^{\circ}C$ and to investigate resuscitation of the VBNC cells in temperatures changed from 10 to $25^{\circ}C$, with and without additives. E. tarda entered into the VBNC state within about 42-84 days of incubation in the microcosm. Throughout this period, the total cell counts as determined using acridine orange direct counting remained near the original inoculum level of ${\sim}10^8cells/ml$. The live cell counts measured with direct viable counting, on the other hands, declined to ${\sim}10^4cells/ml$. When the VBNC cells were incubated with addition of yeast extract, fish muscle extract or serum at $25^{\circ}C$, the ratios of resuscitated samples were 37%, 23%, and 37%, respectively. The characteristics of resuscitated E. tarda were consistent with those of the original E. tarda. When the resuscitated E. tarda were intraperitoneally injected into olive flounders, all fishes died within 5 days, indicating that the VBNC E. tarda might retain its pathogenic potential. Therefore, E. tarda under starvation conditions in the winter enter into the VBNC state and the VBNC E. tarda cells resuscitated at summer and autumn seawater temperature are considered to be pathogen continuously to olive flounder on the southern coast of Korea.
The characteristics of growth and esterase activity of bacterial strains isolated from acid hydrolysed soybean protein were examined. All the isolated strains having decomposition activity of p-hydroxybenzoic acid butyl ester and esterase producing activity were identified as Bacillus sp. by morphological and biochemical methods. The specific growth rates, esterase activities and p-hydroxybenzoic acid butyl ester decomposition activities of isolated strains were $0.844{\sim}1.213\;h^{-1}$, $21{\sim}222\;mU/ml$ and $5.4{\sim}8.1\;mU/ml$, respectively. In the fermentation of Bacillus sp. KB8 strain which had the highest esterase producing activity, growth, extracellular excretion and intracellular synthesis of esterase were inhibited by adding NaCl in the culture broth. Esterase producing activity gradually increased after late exponential growth phase, until maximum value of 420 mU/ml reached after 64 hours culture period. Esterase of Bacillus sp. KB8 strain was stable up to $50^{\circ}C$ for 30 minutes, but was inactivated by heating for 30 minutes at $70^{\circ}C$. The enzyme activity exponentially decreased during the incubation time at the temperatures of $60^{\circ}C$ and $65^{\circ}C$.
Studios on the effects of rice straw amendments, soil autoclaving and repeated application related to disappearance of diazinon (diethyl 2-isopropyl -4-methyl -6-pyrimidinyl phosphorothionate) in submerged soils and paddy water were conducted under the laboratory conditions. Degradation of diazinon was slightly accelerated by the amendment of rice straw. The amended soil had 2.4 days shorter half life for diazinon than unamended soil. By autoclaving soils, diazinon degradation was greatly inhibited. The autoclaved soil had about 20 days longer half life for diazinon than the non-autoclaved soil. After repeated application of diazinon granules to the submerged soils, rapid degradation of the insecticide occured in flooded soils and paddy water. The development of diazinon degrading factors in flooded soils and paddy water after repeated application was roughly proportional to the increase of the frequency of diazinon application. By autoclaving soils and paddy water which received repeated application of diazinon, no rapid biodegradation of the insecticide occurred during the 30 days incubation period.
Song, Tae Hwa;Han, Ouk kyu;Park, Tae Il;Kim, Dae Wook;Yoon, Chang;Kim, Kee Jong;Park, Ki Hun
Journal of The Korean Society of Grassland and Forage Science
/
v.32
no.4
/
pp.335-342
/
2012
This study was conducted to observe the fermentative quality and anthocyanin content in whole crop colored barley silage during storage periods and anthocyanin stability in in vitro ruminal fluid. Silages of colored barley cultivar "Boanchalbori" and normal barley cultivar "Yuyeonbori" were stored during 0, 2, 4, 6, and 12 months. The in vitro ruminal fluid was fermented for 0, 6, 12, 24, and 48 hrs. For the feed value, crude protein of colored barley silage was slightly increased in the silage compared to that of normal barley silage, and being increased up to 2 months after ensiling and thereafter maintained at the similar level. Neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents of both the barley significantly increased by prolonged storage of 2 months (p<0.05), but they were maintained at the constant level after 2 months of storing silage. Whereas TDN (total digestible nutrients) contents of them were decreased by the prolonged storage of 2 months (p<0.05), then maintained at the constant levels. The fermentative quality and pH values in both the barley silages were slightly decreased during the storage time. Lactic acid and acetic acid contents were increased during prolonged storage period, but not significantly different among treatments. Butyric acid was not detected. In the colored barley silage, pH value showed slightly lower compared to that of the normal barley silage but not significant, and lactic acid content was significantly higher than the normal barley silage (p<0.05). The total anthocyanin content in the whole crop colored barley silage decreased to 42% after 2 months of ensilage, however maintained at the constant level until 12 months of ensilage. In the case of anthocyanin stability on in vitro ruminal fluid digestion, the pH value of the ruminal fluid was slightly lower at 6, 12, 24, 48h incubation time and the content of anthocyanin was at similar levels. These results indicated that the colored barley showed higher fermentation quality, and total anthocyanin content was maintained stable at 42% level of the first value in storing silage. As the anthocyanin had higher stability in the ruminal fluid, the colored barley has a potential as functional feeds for Ruminants.
Chun Jae-Woo;Ma Chae-Woo;Kahng Hyung-Yeel;Oh Kye-Heon
Microbiology and Biotechnology Letters
/
v.33
no.2
/
pp.136-141
/
2005
A bench-scale feasibility study was conducted with solid farming feed to evaluate a treatment process for microbiological removal of nitrogen (N) and phosphorus (P). Strains, Bacillus sp. CK-10 and Bacillus sp. CK-13, were originally isolated from water samples of shrimp farming pond. Simultaneous removal of N/P in marine media was monitored in the co-cultures, CK-10 and CK-13. As the results, $400\;{\mu}M\;NH^{+}_4$ and $400\;{\mu}M\;NO^{-}_2$ were eliminated within 12 hours and $NO^{-}_3$ within 36 hours, and $500\;{\mu}M\;PO^{-3}_4$ was completely disappeared within 36 hours from the media. Cultures of CK-10 and CK-13 were applied for removal of N/P leached from shrimp farming fred. HPAEC-PAD system was used to analyze sugars in farming feed, resulting in resolution of various sugars including glucose, galactose, galatosamine, mannose, and fucose. $0.2\%$ (w/v) Pulp densities of the farming feed contained approximately $33.3\;{\mu}M\;NH^{+}_4,\;12.9\;{\mu}M\;NO^{-}_2.\;81.5\;{\mu}M\;NO^{-}_3\;and\;248\;{\mu}M\;PO^{-3}_4$ which could dissolved within 72 hours of leaching in aqueous solution followed by bacterial removal. Complete bacterial removal of N/P was achieved within 84 hours at $0.2\%$ of the feed in co-cultures, whereas single cultures removed to incompletion of N/P during the incubation period. This work demonstrated that test cultures, CK-10 and CK-13 showed effective removal of N/P derived from shrimp farming feed.
Nikfarjam, Bahareh Abd;Adineh, Mohtaram;Hajiali, Farid;Nassiri-Asl, Marjan
Journal of Pharmacopuncture
/
v.20
no.1
/
pp.52-56
/
2017
Objectives: Neutrophils represent the front line of human defense against infections. Immediately after stimulation, neutrophilic enzymes are activated and produce toxic mediators such as pro-inflammatory cytokines, nitric oxide (NO) and myeloperoxidase (MPO). These mediators can be toxic not only to infectious agents but also to host tissues. Because flavonoids exhibit antioxidant and anti-inflammatory effects, they are subjects of interest for pharmacological modulation of inflammation. In the present study, the effects of rutin on stimulus-induced NO and tumor necrosis factor $(TNF)-{\alpha}$ productions and MPO activity in human neutrophils were investigated. Methods: Human peripheral blood neutrophils were isolated using Ficoll-Hypaque density gradient centrifugation coupled with dextran T500 sedimentation. The cell preparations containing > 98% granulocytes were determined by morphological examination through Giemsa staining. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without rutin ($25{\mu}M$) for 45 minutes, and stimulated with phorbol 12-myristate 13-acetate (PMA). Then, the $TNF-{\alpha}$, NO and MPO productions were analyzed using enzyme-linked immunosorbent assay (ELISA), Griess Reagent, and MPO assay kits, respectively. Also, the viability of human neutrophils was assessed using tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and neutrophils were treated with various concentrations of rutin ($1-100{\mu}M$), after which MTT was appended and incubated at $37^{\circ}C$ for 4 hour. Results: Rutin at concentrations up to $100{\mu}M$ did not affect neutrophil viability during the 4-hour incubation period. Rutin significantly decreased the NO and $TNF-{\alpha}$ productions in human peripheral blood neutrophils compared to PMA-control cells (P < 0.001). Also, MPO activity was significantly reduced by rutin (P < 0.001). Conclusion: In this in vitro study, rutin had an anti-inflammatory effect due to its inhibiting NO and $TNF-{\alpha}$ productions, as well as MPO activity, in activated human neutrophils. Treatment with rutin may be considered as a therapeutic strategy for neutrophil-mediated inflammatory/autoimmune diseases.
To evaluate the effects of microorganism agents on oil biodegradation, treatability and microcosm studies were conducted. Petroleum oil degrading bacteria were isolated from enriched cultures of oil-contaminated sediment samples using a mineral salts medium (MSM) containing 0.5% Arabian heavy crude oil as the sole carbon source. After a 5 day-incubation period using MSM, mixed microorganisms of three species (strains BS1, BS2 and BS4) degraded 48.4% of aliphatic hydrocarbons and 30.5% of aromatic hydrocarbons. Treatability and microcosm tests were performed in the three different treatment conditions (AO: Arabian heavy crude oil, AO+IN: Arabian heavy crude oil+inorganic nutrient, AO+IN+MM: Arabian heavy crude oil+inorganic nutrient+mixed microorganism agents). Among these, significantly enhanced biodegradation of aliphatic hydrocarbons were observed in AO+IN and AO+IN+MM conditions, without showing any different biodegradation rates in either condition. However, the degradation rates of aromatic hydrocarbons in an AO+IN+MM condition were increased by 50% in the treatability test and by 13% in the microcosm test compared to those in an AO+IN condition. Taken together, it can be concluded that mixed microorganism agents enhance the biodegradation of aliphatic and aromatic hydrocarbons in laboratory, a treatability test, and a microcosm test. This agent could especially be a useful tool in the application of bioremediation for removal of aromatic hydrocarbons.
Kim, Eun-Ah;Mann, So-Yon;Kim, Su-In;Lee, Ga-Young;Lee, Byong-Won;Kim, Dong-Seob
Journal of Life Science
/
v.24
no.1
/
pp.46-53
/
2014
Lactobacillus sakei 383, which showed the highest GABA content in fermented soycurd, survived in artificial gastric fluid (pH 3.0) up to 3 h, and the survival rate was 88%. L. sakei 383 was tolerant to bile juice during incubation in MRS broth with 0.3% oxgall, and the survival rate was 99%. The survival ratio of L. sakei 383 was high in media containing less than 6% NaCl. L. sakei 383 produced an antibacterial substance against various pathogens, including Listeria monocytogenes, Bacillus cereus, Staphylococcus aureus, Escherichia coli, and Salmonella typhi. The quality characteristics of soycurd fermented with L. sakei 383 were measured during the fermentation period. The viable cell number reached a peak ($10^{11}CFU/ml$) 36 h after fermentation and then slowly decreased. According to the fermentation time of L. sakei 383, the acidity of soycurd increased and the pH decreased until 12 h, and they were maintained thereafter. The moisture, crude ash, crude protein, crude fat, and crude fiber content was 94.88, 0.22, 2.38, 1.16, and 0.03%, respectively. The content of total and reducing sugar was comparatively higher in the soycurd fermented with L. sakei 383 than in nonfermented soycurd. The essential and nonessential amino acid content was 11.2 and 38.65 mg/100 g.
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