• Title/Summary/Keyword: Dark incubation

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The Effect of Temperature and Salinity on Maturation and Hatching of Fibricola seoulensis eggs (온도 및 염도가 Fibricola seoulensis 충란의 성숙과 탈각에 미치는 영향)

  • Lee, Soon-Hyung;Lee, Ho-Jin;Hong, Sung-Tae;Huh, Sun;Seo, Byong-Seol
    • Parasites, Hosts and Diseases
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    • v.24 no.2
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    • pp.115-120
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    • 1986
  • This study was carried out to reveal the effect of temperature, salinity and aeration on maturation and hatching of Fibricola seoulensis eggs. The eggs were incubated and were observed daily for the appearance of eyespots and hatching. The results were summarized as follows. 1. From $4{\sim}5$ days after incubation in distilled water at $28^{\circ}C$ or at $11{\sim}26^{\circ}C$, the eyespots began to appear and the rates of eggs with eyespots were over 90% in $28^{\circ}C$ on the 7th or 8th day. However, eyespots did not appear in $5{\sim}15^{\circ}C$ or $4^{\circ}C$ by the 18th day. 2. The mature eggs began to hatch at the 8th day, and hatching rate 2 weeks after incubation was over 90% at $28^{\circ}C$, but it was below 5% at $11{\sim}26^{\circ}C$, and 0% at $5{\sim}15^{\circ}C$ and at $4^{\circ}C$. 3. Aeration did not influence the appearance of eyes pots nor hatching. 4. In salines under 0.6%, the rates of eyespots appearance were over 90% on the 7th day. The rate was 55.0% in 0.9% at 20 days, and 0% in 1.2%. 5. The hatching rates in salines below 0.3% concentration were over 90% by 14 days of incubation. However, the rate decreased to 44% in 0.6% saline and to 0% over 0.9% salinity. 6. The eggs incubated in the dark hatched in 12.5% on the 10th day, but hatching rate of mature eggs increased to 85.7% within 2 hours after exposure to light. Above results demonstrated that the best temperature for maturation and hatching of F. seoulensis eggs was $28^{\circ}C$, and the miracidia began to hatch at $8{\sim}9$ days after incubation. In the field, hatching and invasion into snails of the miracidia may occur from May to September in Korea. In salines under 0.3% concentration maturation and hatching were not influenced, but as salinity increased hatching was inhibited more than maturation was.

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Long-Term Starvation Induces the Viable-but-Nonculturable Condition in Lactobacillus crispatus KLB46

  • Lee, Seok-Yong;Kim, Ju-Hyeon;Jang, Jeong-Eun;Kim, Seung-Cheol;Yun, Hyeon-Sik;So, Jae-Seong
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.918-922
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    • 2001
  • In a previous study, we have isolated a number of lactobacilli from Korean women, and one of them (KLB46) was identified as Lactobacillus crispatus by 16S rRNA gene sequencing. For the ecological treatment of bacterial vaginosis (BV) cell suspension of L. crispatus KLB46 was instillated into BV patients. L. crispatus KLB46 was found to persist for several days in cell suspension with no nutrients. In this study, in order to assess the influence of starvation on physiological activity, we compared the viability and culturability of KLB46 following suspension in various buffer solutions. A pair of in situ fluorescent dye was used to assess viability (i.e. membrane integrity) and the culturability was examined by plate count assay. A rapid epifluorescence staining method using the LIVE/DEAD Bacterial Viability Kit $(BacLight^{TM})$ was applied to estimate both viable and total counts of bacteria in cell suspension. $BacLight^{TM}$ is composed of two nucleic acid-binding stains ($SYTO\;9^{TM}$ and propidium iodide). $SYTO\;9^{TM}$ penetrates all bacterial membranes and stains the cells green while propidium iodide only penetrates cells with damaged membranes, therefore the combination of the two stains produces red fluorescing cells. Optimal staining conditions for $BacLight^{TM}$ were found to be with 0.0835M $SYTO\;9^{TM}$ and 0.05M propidium iodide for 15 min incubation at room temperature in dark. When cells were microscopically examined during 140 hours of starvation, the culturability decreased markedly while the viability remained relatively constant, which suggests that large fraction of KLB46 cells became viable but non-culturable (VBNC) upon starvation.

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A New Selective Medium for Detecting Acidovorax avenae subsp. avenae in Rice Seeds

  • Song, Wan-Yeob;Kang, Mi-Hyung;Kim, Hyung-Moo
    • The Plant Pathology Journal
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    • v.16 no.4
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    • pp.236-241
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    • 2000
  • A selective agar medium was developed and tested for the isolation of Acidovorax avenae subsp. avenae, the causal bacterial pathogen of bacterial brown stripe, from rice seeds. The new selective agar medium, designated sorbitol pyroglutamic acid agar (SPA) medium, contained 0.5 g of $K_2$HPO$_4$, 3.0 g of Na$_2$HPO$_4$, 2.0 g of D-sorbitol, 0.2 g of L-pyroglutamic acid, 10.0 $m\ell$ of tween 80, 40.0 mg of victoria blue B, 15.0 g of agar, 150.0 mg of ampicillin and 25.0 mg of vancomycin per litter. Colonies of A. avenae subsp. avenae on SPA medium were smooth, round, convex, shiny, blue and 1.5-2.0 mm in diameter 4 days after incubation at 28$^{\circ}C$. Blue colored colony having dark blue zone was typical type of A. avenae subsp. avenae colonies on the medium. Mean recovery of 8 isolates of A. avenae subsp. avenae on the selective SPA medium was 95.8% in comparison to that on KB medium. The saprophytic bacteria were reduced to 97.9% on SPA medium compared to those on KB medium. Most of other rice seedborne bacteria as well as reported pathogenic bacteria were failed to grow on SPA medium. This medium was highly selective for recovering A. avenae subsp. avenae from rice seed samples, and it could be used to enhance the recovery of this bacterium from rice seed samples, which may be contaminated with large numbers of competing microorganisms.

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Cytotoxicity of Shikonin Metabolites with Biotransformation of Human Intestinal Bacteria

  • Min, Byung-Sun;Meselhy, Meselhy-R.;Hattori, Masao;Kim, Hwan-Mook;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.514-517
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    • 2000
  • Abstracts Six shikonin metabolites were obtained from human intestinal bacteria, Bacteriodes fragilis subsp. thetaotus. following biotransformation. The transformation of shikonin (1) was performed anaerobically for 3 day at $37^{\circ}C$ in thc bacterial suspension of B. Fagilis which was cultured overnight in GAM broth. The incubation mixture \vas extracted with EtGAc Lo give a dark-brown residue. The residue was apphed to a silica gel column, which was eluted successively with hexane (Fr. A), $CHCl_3$ (Fr. B), and $CHCl_3$:MeOH (9:I) (Fr. C). Six metabolites. Fr.A (2 and 3), Fr. B (6 and 7), and Fr. C (4 and 5) were isolated by repeated silica gel column chromatography, preparatlVe TLC, followed by Sephadex LH-20. In vitro cytotoxicities were tested against human tumor cell lines; PC-3 (prostate), ACHN (renal), A549 (lung), SW620 (colon), KS62 (leukemia), and Du145 (prostate). The shikonin metabolites 2. 4, 5, and 6 showed weaker cytotoxicity than the parenL shikonin (1). whereas shikonin monomenc metabolite 3 ($ED_{50}{\;}O.44-{\;}1.22{\;}\mu\textrm{g}/ml$) and dimeric metabolite 7 ($ED_{50}{\;}O.48-{\;}2.35{\;}\mu\textrm{g}/ml$) exhibited stronger activities compared with adriamycin, which was used as the positive control.ontrol.

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Changes in Dissolved Organic Matter Composition in the Namhan River during a Heavy Rain Event (집중 강우시 남한강 내 용존 유기물의 성상 변화)

  • Oh, Seijin;Woo, Sungho;Hur, Jin;Jung, Myung-Sook;Shin, Hyun-Sang
    • Journal of Korean Society on Water Environment
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    • v.25 no.5
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    • pp.697-703
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    • 2009
  • In this study, changes in the composition of dissolved organic carbon (DOC) were investigated using water samples collected at a downstream site of the Namhan River near the Lake Paldang ($N37^{\circ}24^{\prime}05.33^{{\prime}{\prime}}E127^{\circ}32^{\prime}25.01^{{\prime}{\prime}}$) during a heavy rain event from July 23 to July 28, 2008. The DOC concentrations varied from 1.68 to 3.18 mg/L with the maxium value at a peak of the river water level. Each DOC sample was fractionated into three compositions including hydrophilic (Hi), hydrophobic acid (HoA) and hydrophobic neutral (HoN) fractions. The results showed that HoA was most abundant fractions, constituting 47.2~56.5% of DOC. Refractory dissolved organic carbon (R-DOC) contents were also determined by measuring the DOC concentration after 28-day dark incubation of the samples. R-DOC content was in the range from 83 to 99% of DOC and it was significantly correlated with HoA contents (r = 0.91, p<0.005), while it did not exhibit such a good correlation with the fractions of Hi and HoN (p>0.02). Our results suggest that the HoA, which is associated with humic substances, may be a major composition of refractory organic matters in rivers during storm events.

Evaluation of Algal Growth Potential in the Mangyeong River by MBOD method (MBOD법에 의한 만경강 수계의 조류성장잠재력 평가)

  • Kim, Jong Gu;Kim, Jun U
    • Journal of Environmental Science International
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    • v.13 no.9
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    • pp.807-817
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    • 2004
  • The modified biochemical oxygen demand (MBOD) were conducted to evaluate the water quality and fertility in the Mangyeong river from november 2002 to april 2003. MBOD method was used to evaluate algal growth potentials and their limiting factors. MBOD depends on the amount of available inorganic nutrient and organic substrate during 5-day incubation in the dark condition at $20^{\circ}C.$ The MBOD assay depends on inorganic nutrients such as phosphorus and nitrogen as well as reduced carbon as called MBOD, MBOD-P, and MBOD-N, respectively. The concentration of pollutants were in the range of 3.08~48.36 mg/L for COD. The concentration of nutrients were in the range of 0.37~111.62 mg/L for dissolved inorganic nitrogen (DIN) and 0.00~1.03 mg/L for dissolved inorganic phosphorus (DIP). The results of MBOD bioassay showed that the MBOD, MBOD-P and MBOD-N values were 15~173 mg $O_2/L,$ 13~165 mg $O_2/L$ and 66~175 mg $O_2/L$ ranges, respectively. The MBOD values are found to be the highest in Iksan River and the lowest in Hari River throughout the Mangyeong River. The relationships of MBOD, MBOD-P and MBOD-N in MBOD method were generally found in MBOD$\risingdotseq$ MBOD-P$\risingdotseq$MBOD-N. But the result of Gosan was appeared to MBOD$\risingdotseq$MBOD-N > MBOD-P. The MBOD-N value was higher 3 to 5 times than the MBOD-P value in the Gosan station. The algal growth potentials expressed as the concentration of chlorophyll-a were maximum 20 times more than algal biomass in the water column.

CELLULAR ATTACHMENT AND GENE EXPRESSION OF OSTEOBLAST-LIKE CELLS ON ZIRCONIA CERAMIC SURFACES

  • Pae, Ah-Ran;Lee, Hee-Su;Kim, Hyeong-Seob; Baik, Jin;Woo, Yi-Hyung
    • The Journal of Korean Academy of Prosthodontics
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    • v.46 no.3
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    • pp.227-237
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    • 2008
  • STATEMENT OF PROBLEM: Zirconium oxide can be a substitute to titanium as implant materials to solve the esthetic problems of dark color in the gingival portion of implant restorations. PURPOSE: This study was performed to define attachment and growth behavior of osteoblast- like cells cultured on grooved surfaces of zirconium oxide and evaluate the genetic effect of zirconium oxide surfaces using the reverse transcriptase-polymerase chain reaction (RT-PCR). MATERIAL AND METHODS: MC3T3-E1 cells were cultured on (1) commercially pure titanium discs with smooth surface (T group), (2) yttrium-stabilized tetragonal zirconia polycrystal (Y-TZP) with machined surface (ZS group), and (3) Y-TZP with $100{\mu}m$ grooves (ZG group). Cell proliferation activity was evaluated through MTT assay and cell morphology was examined by SEM. The mRNA expression of Runx2, alkaline phosphatase, osteocalcin, TGF-${\beta}1$, IGF-1, G3PDH in E1 cells were evaluated by RT-PCR. RESULTS: From the MTT assay, after 48 hours of adhesion of MC3T3-E1 cells, the mean optical density value of T group and ZG group significantly increased compared to the ZS group. SEM images of osteoblast-like cells showed that significantly more cells were observed to attach to the grooves and appeared to follow the direction of the grooves. After 24 hours of cell adhesion, more spreading and flattening of cells with active filopodia formation occurred. Results of RT-PCR suggest that T group, ZS group, and ZG group showed comparable osteoblast-specific gene expression after 24 hours of cell incubation. CONCLUSION: Surface topography and material of implants can play an important role in expression of osteoblast phenotype markers. Zirconia ceramic showed comparable biological responses of osteoblast-like cells with titanium during a short-time cell culture period. Also, grooves influence cell spreading and guide the cells to be aligned within surface grooves.

Adaptations of Estuarine and Freshwater Phytoplankton to Urea Decomposition (기수 및 담수 식물플랑크톤의 요소 분해에 대한 적응)

  • PARK, MYUNG GIL;SHIM, JAE HYUNG;CHO, BYUNG CHEOL
    • 한국해양학회지
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    • v.28 no.4
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    • pp.323-331
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    • 1993
  • The concentration-dependence of and the effect of light on urea decomposition, and the suppression of urea decomposition by ammonium were studied to understand adaptations in phytoplankton to utilization of urea in the estuarine system of the Mankyung and Dongjin rivers and a hypertrophied pond. Results of size-fractionation showed that bacterial fraction played a minor role (14%) in urea decomposition in the estuary. However, the role of bacteria in urea decomposition seemed to increase in a hypertrophic pond. Natural phytoplankton communities exhibited a monophonic or biphasic kinetics of urea decomposition over a wide range of concentration (upto 7.7 mM). the addition of high concentration of ammonium and incubation of the euphotic samples in the dark caused reductions in the urea decomposition rates. It is suggested that understanding of adaptations in phytoplankton to urea decomposition would help to study the temporal and spatial variabilities of urea decomposition rates in the field and the significance of urea in nitrogen cycle.

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Seed-borne Infection of Alternaria sesami and Corynespora cassiicola in Sesame, Damage to Seeds and Seedlings and Control by Seed Treatment (참깨의 종자전염성(種子傳染性) Alternaria sesami와 Corynespora cassiicola에 관(關)하여 - 종자(種子) 및 유묘(幼苗)에 미치는 피해(被害)와 종자소독(種子消毒)의 효과(效果) -)

  • Yu, Seung Hun
    • Korean Journal of Agricultural Science
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    • v.10 no.2
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    • pp.269-276
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    • 1983
  • Out of 50 seed samples of Sesamum indicum L. tested, Alternaria sesami and Corynespora cassiicola were detected in 29 samples and 38 samples, respectively. Heavy infection of A. sesami and C. cassiicola caused seed rot and seedling blight on water agar as well as in pots. In case of light infection the seed germinated showing elongation of the radicle and hypocotyl, but soon light brown or dark brown lesions appeared at certain parts of the shoot and root where sporulation of the fungi was observed after prolonged incubation in moisture. Four seed disinfectants were compared for the control of seed-borne infection of A. sesami and C. cassiicola. Complete control of seed-borne infection of C. cassiicola was obtained by seed treatment with Benlate T and Homai and seed treatment with Busan 30 was found superior to control seed-borne infection of A. sesami.

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Underdeveloped Embryos and Dormancy Type in Seeds of Two Heloniopsis Species Endemic to Korea

  • Lee, Seung Youn;Rhie, Yong Ha;Kim, Ki Sun
    • Horticultural Science & Technology
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    • v.32 no.4
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    • pp.550-557
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    • 2014
  • Heloniopsis koreana and Heloniopsis tubiflora (Melanthiaceae) are endemic herbaceous species of the Korean Peninsula. The Melanthiaceae family has been described as having seeds with small, underdeveloped embryos at the time of dispersal, and morphological (MD) or morphophysiological dormancy (MPD). However, there are few reports on embryo growth, morphology, and seed germination in Heloniopsis species. The aims of this study were to investigate embryo growth and seed dormancy, and to determine the type of dormancy exhibited by these species. The effects of incubation temperatures, light conditions, and gibberellic acid ($GA_3$) on dormancy break and seed germination were tested. Freshly matured seeds of the two species had small embryos that occupied about 9-11% of the length of the endosperm, and which increased by more than 300% in length before radicle emergence, indicating that the embryos are underdeveloped at the time of dispersal. Embryos in the seeds grew under warm temperature regimes (between $25/15^{\circ}C$ and $30/20^{\circ}C$). $GA_3$ application (tested only in the light) overcame seed dormancy and promoted germination. Approximately 30% of the seeds of H. koreana and approximately 40% of the seeds of H. tubiflora germinated in suitable environmental conditions (light and temperature) within 4 weeks. Therefore, 30-40% of the seeds of the two species exhibited MD, and the rest of the seeds had non-deep simple MPD. Light was found to be one of the critical factors for germination because no seed of either of the two Heloniopsis species germinated under constant dark conditions, and thus, these species have the potential to form a persistent soil seed bank. Understanding these germination requirements will help in development of effective strategies to increase the establishment of seedlings in their native habitat.