In developing soil wetting agent using polyoxyethylene nonylphenyl ether (PNE) and polyoxyethylene castor oil (1:1; v/v), the effect of application rates on changes in concentration of PNE, initial wetting of peatmoss + perlite (7:3) medium, and growth of hot pepper (Capsicum annuum L. 'Knockwang') plug seedlings were investigated. The elevation of application rates of wetting agent increased the amount of water retained by the root media. The treatment of 2.5 $mL{\cdot}L^{-1}$ showed similar water retention to + control ($AquaGro^L$ 3.0 $mL{\cdot}L^{-1}$). Most of the liquid wetting agent (LWA) incorporated during the medium formulation leached out in the first and second irrigation, then it decreased gradually until 10 times in irrigation. In investigation of the influence of LWA on position of water infiltrating into root media, the vertical water movements in treatments of 0.5, 1.0, and 1.5 $mL{\cdot}L^{-1}$ were much faster than those in 0.0 $mL{\cdot}L^{-1}$ (-control), but relative speed of water movement decreased by the elevation in application rate of LWA to 2.0 or 2.5 $mL{\cdot}L^{-1}$. The evaporative water loss of root media that to contained various rate of LWA and irrigated to reach container capacity was the fastest in -control among the treatments and it delayed as the application rate of LWA was elevated. The plant height of 22.2 cm in 0.5 $mL{\cdot}L^{-1}$ and stem diameter of 3.26 mm in 1.0 $mL{\cdot}L^{-1}$ were the highest among the treatments tested. The treatment of 1.0 $mL{\cdot}L^{-1}$ also had the heaviest fresh and dry weights such among treatments tested as 3.08 g and 0.861 g per plant, respectively. The elevated application rate over than 1.5 $mL{\cdot}L^{-1}$ resulted in decreased seedling growth. The results mentioned above indicate that optimum application rate of LWA is 1.0 $mL{\cdot}L^{-1}$.
1. Isolation and identification of amylase-producing bacteria. The powerful strain A-12 and S-8 were respectively isolated from air and soil after screening a large number of amylase-producing bacteria. Their bacterial characteristics have been investigated and it has been found that all characteristics of strain A-12 and S-8 are similar to Bac. subtilis of Bergey's manual except for the acid formation from a few carbohydrates and the citrate utilization, i.e., the strain A-12 shows negative in the citrate utilization, and the acid formation from arabinose and xylose, S-8 shows negative in the acid formation from xylose. 2. Amylase production by Liquid cultures with solid materials. Several conditions for amylase production by strain A-12 in stationary cultures have been studied. The results obtained are as follows. (1) The optimum conditions are:temperature $35^{\circ}C$, initial pH 6.5 to 7.0 and incubation time 3 to 4 days. (2) The amylase production is not affected by the preservation period of the stock cultures. (3) Among the various solid material, the defatted soy bean is found to be the best for t1e amylase production. However, the alkali treatment of the defatted soy bean gives no effect contrary to the cage of defatted rape seed. The addition of soluble starch to the alkali extract of defatted soy bean shows the increased amylase production. (4) Up to 1% addition of ethanol to carbon dificient media gives the improved amylase production, whereas the above effect is not found in the case of carbon rich media. (5) The amylase production can be increased 2.5 times when 10% of defatted soy bean is admixed to cheaply available wheat bran. (6) The excellent effect is found for amylase production when 20% of wheat bran is admixed to defatted dry milk which is a poor medium. The activity is found to be $D^{40^{\circ}}_{30'}$ 7,000(L.S.V. 1,800) in 10% medium. (7) No significant effect is observed due to the addition of various inorganic salts. 3. Amylase production by solid cultures. Several conditions for amylase production by strain A-12 in wheat bran cultures have been studied and the results obtained are as follows. (1) The optimum conditions: are temperature $33^{\circ}C$, incubation lime 2 days, water content added 150 to 175% and the thickness of the medium 1.5cm, The activity is found to be $D^{40^{\circ}}_{30'}$ 36,000(L.S.V. 15,000) (2) No significant effect is found in the case of the additions of various organic and inorganic substances.
Jo, Eun Ju;Lee, Ji Hyun;Choi, Yong Ho;Kim, Jin-Cheol;Choi, Gyung Ja
Horticultural Science & Technology
/
v.33
no.3
/
pp.409-419
/
2015
This study was conducted to establish an efficient screening method for watermelon plants resistant to Fusarium wilt (FW), which is caused by Fusarium oxysporum f. sp. niveum (Fon). An HA isolate was prepared from a wilted watermelon plant in Haman-gun and identified as F. oxysporum f. sp. niveum based on morphological characteristics, molecular analyses of ITS (internal transcribed spacer) and TEF (translation elongation factor $1{\alpha}$) sequences, and host specificity on cucurbits including watermelon, melon, oriental melon, and cucumber. The assay for disease response of watermelon differentials indicated that the HA isolate was race 0. Among seven liquid media tested, the highest amount of Fon spores was produced from V8-juice broth, which was selected as a medium for mass production of Fon. The disease assay for 21 watermelon and 11 watermelon-rootstock cultivars demonstrated that 20 watermelon cultivars except for 'Soknoranggul' were susceptible; 'Soknoranggul' was moderately resistant. All the tested rootstock cultivars were highly resistant to the HA isolate. The evaluation of disease development depending on various conditions suggested that an efficient screening method for FW resistance in watermelon plants is to dip the roots of 10-day-old seedlings in spore suspension of $1.0{\times}10^5-1.0{\times}10^6conidia{\cdot}mL^{-1}$ for 30 min., to transplant the seedlings to plastic pots with a fertilized soil, and then to cultivate the plants at $25^{\circ}C$ for 3 weeks.
The quality characteristics of Takju were investigated according to the different rice varieties and mixing ratio of Nuruk for the advanced quality of Takju. The yeast was selected by alcohol-producing ability. Then a liquid starter was prepared using brewing fungi (Aspergillus luchuensis 34-1 and Lichtheimia ramosa CN042), and rice Nuruk was manufactured with two rice types (Chucheong and Hanareum). The quality characteristics of Takju were investigated based on the rice type and the mixing ratio of A. luchuensis 34-1 and Lich. ramosa CN042 (1:0, 0:1, 1:1, 1:3). S. cerevisiae Y268 showed an alcohol yield of $9.3{\pm}0.33%$ at a 0.3% concentration in the YPD broth medium, and the rice Nuruk with A. luchuensis 34-1, regardless of rice type, was confirmed to have a higher enzyme activity and physiochemical property than Lich. ramosa CN042. According to the quality analysis of Takju, the physiochemical property was increased for the fermentation period, and the acidity differed by type of fungi and rice. The quality of Takju was changed with the composition differences of organic acid and free amino acid by rice type and mixing ratio. As a result of the sensory evaluation of Takju, the preference for it was increased with the Hanareum and A. luchuensis 34-1, respectively. Thus, this study shows the possibilities for activating the industry of traditional liquor by improving the Nuruk and Takju manufacturing technique.
Kim, I.-D.;Ahn, M.-H.;Hur, T.-Y.;Hong, M.-P.;Seok, H.-B.
Journal of Embryo Transfer
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v.19
no.2
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pp.155-163
/
2004
The aims of this study are 1) to test oocytes and embryos collected from in-vitro to achieving the valuable protocol by culturing, vitrifying and thawing of oocytes/embryos, and 2) to transfer them to recipient, and finally have resulted in pregnancies from recipient females after surgical or nonsurgical transfer. In vitro maturation and fertilization were performed according to Funahashi et al (1994). Glucose-free NCSU 23 supplemented with 5 mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$, and 10% fetal bovine serum albumin was added to the culture medium thereafter. Embryos were treated with 7.5 ${\mu}g/ml$ cytochalasin-B for 30 min, centrifuged at 13,000 rpm for 13 min and then exposed sequentially to an ethylene glycol(EG) vitrification solution, aspirated into OPS, and plunged/thawed into/from liquid nitrogen. In vivo embryos were surgically collected from three dornors after AI for control group. Forty-nine embryos were washed 3 times in mPBS + 10% FBS, followed treatments : cultured, centrifuged, vitrified, recovered and transferred to recipients as in vitro prepared embryos. Three recipients were transferred individually with 100, 100 frozen embryos derived from abattoir and 34 fresh embryos by surgically, and another three recipients were transferred individually with 150, 150 frozen embryos and 100 fresh embryos by nonsurgically, respectively. all recipient sows exhibited delayed returns to estrus. To our knowledge, theses results suggest that required an improved techniques, more vigorous embryos preparation and substitute to gilt with cleaner uterous condition.
Lee Jung-Won;Shin Jee-Sun;Seo Jeong-Wan;Lee Mi-Ae;Lee Seung-Joo
Childhood Kidney Diseases
/
v.8
no.2
/
pp.214-222
/
2004
Purpose: Appropriate antibiotic therapy is important in childhood urinary tract infection and the selection of anibiotics is based on antimicrobial sensitivity of Escherichia coli. Extended-Spectrum ${\beta}-Lactamase(ESBL)$ is an enzyme produced by gram-negative bacilli that has the ability to hydrolyse penicillins, broad-spectrum cephalosporin and monobactam. There have been many reports of outbreaks of hospital infection by ESBL-producing organism. However, community-acquired infection with ESBL-producing organism are rare. This study was performed to retrospectively identify the incidence, characteristics and risk factors of ESBL (+) E. coli in community-acquired childhood UTI. Methods: In 288 children admitted in Ewha Womans University Hospital with E. coli UTI from Mar 2001 to February 2003, ESBL was isolated. ESBL was confirmed by the utilization of an automatized machine(Vitek GNS 433 card) using liquid medium dilution method according to National Committee for Clinical Laboratory Standard. The clinical characteristics, risk factors, antimicrobial resistance and treatment effectiveness were compared with ESBL(-) E. coli UTI. Results: Of 288 E. coli isolates, 31(10.8%) produced ESBL and 93.5%(29/31) occurred in infants younger than 6 month of age(P<0.01). No significant differences were noted in prior antibiotic use, prior admission history and underlying urogenital anomaly. Antimicrobial resistance was significantly higher in ESBL(+) E. coli compared with control patients (P<0.05). Although ceftriaxone showed 100% resistance in ESBL(+) E. coli, bacteriologic sterilization rate after ceftriaxone therapy was higher(96.8%). However, the recurrence rate of febrile UTI within 6 months was higher(25.8%) than control patients(6.6%). Conclusion: Epidemiologic study is required to find out any new risk factors of community-acquired ESBL(+) E. coli UTI and changes in selection of empirical antibiotics should be considered.
Strain A-3, an amylase-producing bacteria, was isolated from coastal seawater near Daecheon in the Republic of Korea. It was seen to possess a single polar flagella and grow well, on ASW-YP agar plates, at temperatures of between $20-37^{\circ}C$. However, it grew more slowly at the temperatures of $15^{\circ}C$ and $40^{\circ}C$. Similarly, it was observed to grow abundantly, in an Artificial Sea Water-Yeast extract-Peptone (ASW-YP) liquid medium, in a pH range of 6-9, but not grow at pHs of 4-5 and a pH of 10. Strain A-3 was noted as being close to Pseudoalteromonas phenolica O-$BC30^T$, Pseudoalteromonas luteoviolacea $NCIMB1893^T$, Pseudoalteromonas rubra $ATCC29570^T$, and Pseudoalteromonas byunsanensis $FR1199^T$, with 98.30%, 97.86%, 97.78%, and 97.25% similarities respectively, in its 16S rRNA sequence. A phylogenetic tree revealed that strain A-3 and P. phenolica O-$BC30^T$ belong to a clade. However, strain A-3 differed from P. phenolica O-$BC30^T$ in relation to a number of physiological characteristics. Strain A-3 exhibited no growth above 5% NaCl concentrations, no utilization of D-glucose, D-mannose, D-maltose, or D-melibose, and no lipase (C-14) activity. All of these properties strongly indicate that strain A-3 is distant from P. phenolica O-$BC30^T$ and thus led us to name it Pseudoalteromonas sp. A-3. Pseudoalteromonas sp. A-3 produces ${\alpha}$-amylase throughout growth. Maximal amylase activities of 144.48 U/mL and 149.20 U/mL were seen at pH 7.0 and $37^{\circ}C$, respectively. Pseudoalteromonas sp. A-3's high, stable production of ${\alpha}$-amylase in addition to its biochemical features, such as alkalitolerance, suggest that it is a good candidate for industrial applications.
The present study was undertaken to investigate the post-thawed survivability of bovine embryo depending on different dose of ethylene glycol and sucrose. Ovaries were collected at local slaughterhouse and the cumulus-oocyte-complexes aspirated from ovaries were in vitro matured, fertilized and cultured at 39°C in an atmosphere of 5% CO2 incubator. For conventional slow-freezing, d 7 or 8 expanded blastocysts were collected. Embryos were equilibrated in 1.5 M and 1.8 M ethylene glycol(EG) with 0.1 M and 0.3 M sucrose in Dulbecco's phosphate-buffered saline(D-PBS) supplemented with 0.5% bovine serum albumin. Embryos were then loaded individually into 0.25ml-straw and placed directly into cooling chamber of programmable freezer precooled to 7°C, after 2 min, the straw was seeded, maintained at 7°C for 8 min, and then cooled to 35°C at 0.3°C/min, plunged and stored in liquid nitrogen for at least 3 days. For thawing, the straw containing embryos were warmed in air for 10 sec and exposed to 37°C water for 20 sec. Straws were then removed from 37°C water. Rates of blastocyst survive and hatching were evaluated at 24 to 72 h post-warming. No difference of the survivability was shown between 1.5 M and 1.8 M EG (71 and 70%, respectively). Addition of 0.1 M sucrose to 1.5 M and 1.8 M ethylene glycol in the freezing solution did not differ significantly embryo survival (74 and 77%, respectively), whereas survival rates was higher(89%) in freezing solution contained 0.3M sucrose to 1.8M EG compared with 0.3M sucrose to 1.5M EG group(71%). However, there was no difference in the overall total cell number between the two groups (122±1.8 vs 131±1.4, respectively). In conclusion, the results suggest that 0.3 M sucrose in 1.8 M EG may be optimal condition for freezing and thawing methods with in vitro produced embryos and may be applied to on-farm conditions for embryo transfer.
Three slime-forming lactic acid bacteria were isolated from traditional Korean fermented soy sauce and soybean paste and shown to produce exopolysaccharides (EPS) in sucrose media. By isolating the strains, examining their morphological characteristics and determining their 16S rDNA sequences, N58-5 and K6-7 were identified as Leuconostoc mesenteroides and N45- 10 as Leuconostoc citreum. The acid and bile tolerances of these three strains were investigated. Amongst the three lactic acid bacteria, Leuc. citreum N45-10 exhibited the highest viability ($10^5-10^6$ CFU/ml) in 0.05 M sodium phosphate buffer (pH 0.3) for 2 h, in artificial gastric juice for 2 h and in 0.3%, 0.5% oxgall for 24h. Leuc. mesenteroides K6-7, N58-5 and Leuc. citreum N45- 10 were grown in sucrose liquid medium and 8.16 g/L, 3.65 g/L, 16.17 g/L of EPS was collected, respectively. The hydrolyzed EPS was analyzed by HPLC in order to determine the sugar composition of EPS. Leuc. mesenteroides K6-7 and N58-5 showed two peaks indicating glucose and fructose, thus they were determined to be hetero-type polysaccharides. Leuc. citreum N45-10 showed only the glucose polymer, indicating it to be a homo-type polysaccharide. In addition, all three lactic acid bacterial hemolysis did not demonstrate a clear zone in blood agar in the area surrounding a lactic acid bacteria colony.
The mycelia of Pyricularia oryzae were treated with enzyme solution mixture consisting of Driselase, ${\beta}-Glucuronidase$, Cellulase, and Macerozyme R-10 for the production of protoplasts. More protoplasts were formed from mycelia of race KJ 101 of P. oryzae than that of race KI 315a in the enzyme mixtures. The number of protoplasts was decreased in the untreated control three hrs after the enzyme treatment, whereas the number was increased in the treatments with 10, 50 and 100 mM 2-mercaptoethanol, respectively. The number of protoplasts increased to reach maximum at five hrs after treatment of 10 mM 2-mercaptoethanol, but the least was found in 200 mM. The protoplasts of P. oryzae in a liquid medium containing 2.5% yeast extract, and 2% dextrose reverted to the mycelia after five hrs shaking incubation at $27^{\circ}C$. Some protoplasts produced yeast like buds and the buds were developed to irregularly shaped chains of cell protruded a germ tube like hypha from the distal cell. Once in a while a germ tube like hypha protruded directly from the protoplasts. Except in the first type of reversion, other protoplasts reverted to the normal mycelia. The reversion frequency was highest on PDA with stabilizer of 0.6 M KCl. No reversion of protoplasts occurred on water agar regaardless oftreaatments.
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