• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.30-41
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• 2011

The influences of the agitation as well as the addition of medium during culture on the production of embryos were invested in isolated microspore culture of hot pepper (Capsicum annuum L.). When the culture medium was added during initial liquid culture step of liquid-double layer culture, the embryo yield and quality greatly increased. The most effective time point for medium addition was 5 days after the culture commenced. On the other hand, the effect of medium addition at later double layer culture step in liquid-double layer culture on the embryo production was less compared to that of medium addition during the initial liquid culture step. Agitating the culture for 1 week during later double layer culture step in liquid-double layer culture effectively increased the production of normal cotyledonary embryos. In the case of liquid culture, agitating the culture for 1 week from 7 days after the culture commenced was also effective for embryo development. However, when the total agitation time was longer (2 to 3 weeks) during liquid-double layer culture or liquid culture, the embryos developed abnormally in both cases. The normal cotyledonary embryos obtained in this study successfully developed to plants when transferred to regeneration media. These regenerated plants were either diploid or haploid, and there was a difference in the number of chloroplasts between guard cells of diploid and haploid. These results can be used as an important data for developing an efficient microspore culture system with high quality embryo production in hot pepper.

• International Journal of Oral Biology
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• v.39 no.1
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• pp.35-40
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• 2014

Halitosis is caused by consumption of certain foods or drinks and production of volatile sulfur compounds (VSCs) by periodontopathogens. VSCs-related halitosis is not easily removed using mechanical or chemical therapies such as dental floss, plaque control and mouth rinse. Lactobacillus are known to be probiotics and stimulate immune systems of human. Furthermore, L. casei ATCC 334 and L. rhamnosus GG have an effect on protection of dental caries in vitro studies. The aim of this study was to investigate effect of Lactobacillus on halitosis by Fusobacterium nucleatum- and Porphyromonas gingivalis-producing VSCs and to analyze inhibitory mechanism. The periodontopathogens were cultivated in the presence or the absence Lactobacillus, and the level of VSCs was measured by gas chromatograph. For analysis of inhibitory mechanisms, the susceptibility assay of the spent culture medium of Lactobacillus against F. nucleatum and P. gingivalis was investigated. Also, the spent culture medium of Lactobacillus and periodontopathogens were mixed, and the emission of VSCs from the spent culture medium was measured by gas chromatograph. L. casei and L. rhamnosus significantly reduced production of VSCs. L. casei and L. rhamnosus exhibited strong antibacterial activity against F. nucleatum and P. gingivalis. The spent culture medium of L. casei inhibited to emit gaseous hydrogen sulfide, methyl mercaptan and dimethyl sulfide from the spent culture medium of periodontopathogens. However, the spent medium of L. rhamnosus repressed only dimethyl sulfide. L. casei ATCC 334 may improve halitosis by growth inhibition of periodontopathogens and reduction of VSCs emission.

• KSBB Journal
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• v.19 no.6 s.89
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• pp.451-456
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• 2004

The effects of variation in composition of the medium on the conversion of Gluconacetobacter hanseii PJK cells producing cellulose ($Cel^+$) to non-cellulose producing ($Cel^-$) mutants and the production of bacterial cellulose (BC) in an agitated culture were investigated. The impeller speed greater than 500 rpm was required to decrease the population of $Cel^-$ mutants to minimum in a basal medium containing $1.5\%$ ethanol because the optimum impeller speed to minimize the population of $Cel^-$ mutants increased with the concentration of ethanol added to a basal medium. Ethanol fed-batch culture could not increase the BC production in an agitated culture unlike that of a shaking culture. The amount of BC produced in a basal medium containing $1\%$ ethanol was $39\%$ more than that of the same medium with $0.27\%\;Na_{2}HPO_4$. Increase in the concentration of acetic acid in a basal medium decreased the BC production. The pH control of the culture broth increased the cell mass in the batch culture and improved the production yield of water-soluble polysaccharide (WSPS), but did not affect the production of BC.

• FLOWER RESEARCH JOURNAL
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• v.17 no.1
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• pp.9-14
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• 2009

This study was conducted to find out the commercial in vitro propagation methods through protocom-like bodies (PLB) induced from shoot tip culture of Miltonia spp. Among several culture media for induction PLB from shoot tip in Miltonia spp., MS basal medium was better than Hyponex, Vacin & Went basal medium and other media supplemented with natural additives. PLB's proliferation and differentiation in Hyponex medium including $20g{\cdot}L^{-1}$ banana + $20g{\cdot}L^{-1}$ sucrose(pH 5.2) was better than in MS medium. It was tendency that solid media showed higher PLB fresh weight than liquid medium or other cotton bridge culture. The dark culture for 1~2 weeks and adding $10{\sim}20g{\cdot}L^{-1}$ sucrose on hyponex basal medium was the most effective to increase the PLBs growth and shoot number.

• Reproductive and Developmental Biology
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• v.31 no.4
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• pp.261-265
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• 2007

Insulin, transferrin and selenium (ITS) complex is reported to improve in vitro development of oocytes and embryos. This study was carried out to investigate the effects of ITS during in vitro culture (IVC) of porcine parthenogenetic and nuclear transfer (NT) embryos on subsequent developmental capacity in vitro. The electrically activated oocytes were cultured in Porcine Zygote Medium (PZM-3) with various concentrations (0, 0.1, 0.5, and 1.0%) of ITS for 7 days. Also, the electrically activated reconstructed embryos were cultured in PZM-3 with various concentrations (0, 0.1, 0.5, and 1.0%) of ITS for 6 days. Addition of ITS to culture medium did not affect development of porcine parthenogenetic embryos in vitro. To test the effect of ITS on the in vitro development of porcine NT embryos, factorial experiments were also performed for in vitro maturation (IVM) medium (TCM-199) with or without 1% ITS and culture medium (PZM-3) with or without 0.5% ITS. Addition of 0.5% ITS to culture medium increased (p<0.05) the proportion of NT blastocysts compared with non-treated group. In contrast, addition of 1% ITS to culture medium was ineffective or had a detrimental effect. Also, addition of ITS only to maturation medium increased (p<0.05) the percentage of NT blastocysts formation compared with the control group. In conclusion, addition of ITS to IVM or IVC medium could improve subsequent blastocyst development of porcine NT embryos.

• Korean Journal of Animal Reproduction
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• v.17 no.2
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• pp.121-125
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• 1993

Early development of bovine oocytes fertilized in vitro in the medium with caffeine and heparin was examined in different culture systems. When the oocytes were transferred into culture medium 8 h after insemination, 12%(7/60) of penetrated oocytes cleaved to 4-cell stage 24 h after insemination. The proportions of oocytes cleaved to 80to 16-cell stage 48 h after insemination had also a to be higher in oocytes transferred into culture medium 8 h (29%) than 16 h(10%) or 24 h(4%) after insemination. 52% of the 4-cell embryos developed to morula and blastocyst stages when they were co-cultured with oviductal epithelia, whereas only 5% of embryos cultured without the epithelial cells(P<0.001). In another experiment, embryos were co-cultured with ampulla, isthmus or utero-tubal junction of oviducts. There are no significant differences in the proportions of embryos developed to morula and blastocyst stage.

• Microbiology and Biotechnology Letters
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• v.25 no.3
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• pp.277-285
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• 1997

Kluyveromyces marxianus var. marxianus isolated as an inulin-assimilating microorganism produces inulin fructotransferase (inulaseII) which catalyses the conversion of inulin into di-D-fructofuranose 1, 2' : 2, 3' dianhydrde (DFAIII). The DFA produced by the organism was isolated by using active carbon column, and identified as DFAIII by high performance liguid chromatography. The culture medium giving maximum inulaseII production was found to consist of 1% sucrose and 0.75% yeast nitrogen base (YNB). The inulasell production was induced by inulin or sucrose as a carbon source and increased by addition of YNB as a nitrogen source. Optimal initial pH of the culture medium, culture temperature and medium volume for the enzyme production were pH 4.7, 30$\circ$C and 140 ml, respectively. Under the optimal conditions described above, the enzyme activity in the culture supematant reached 4.2 units/ml after cultivation for 36 h. The DFAIII was accumulated at 13.25 mg/ml after 48 h of culture in the Jerusalem artichoke tuber medium.

• Parasites, Hosts and Diseases
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• v.38 no.3
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• pp.173-175
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• 2000

The present study examined postmetacercarial changes in the excysted metacercariae of Echinostoma caproni maintained in the defined medium Mixture 199 plus 20% calf serum for 7 days at $41^{\circ}C$. The gas phase was atmospheric air. Each culture was inoculated with 25 excysted metacerariae. Cultures were maintained upright in closed 15 ml plastic centrifuge tubes each containing 10 ml of medium plus 200 units of penicillin/ml and $200{\;}\mu\textrm{g}$ of streptomycin/ml. By 4 days in culture, most metacercariae had voided their excretory concretions. Organisms were clumped or solitary at the bottom of the cultures. Many organisms showed flaring of the oral collar and extension of both the collar and tegumentary spines. By 4 days in culture, posterior protuberances or bumps were noted on many of the organisms and some organisms showed abnormal vesicular growths or blebs at their posterior ends. Some mortality was noted in culture by day 5, but most organisms were still alive when the cultures were terminated on day 7.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.10
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• pp.64-71
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• 2020

This study evaluated the effects of the culture media and light sources on the growth of microalgae Haematococuus pluvialis. Limited ingredient medium, Modified Bold's Basic Medium (MBBM), commercial liquid fertilizer medium Neo, and seven different light sources with different wavelengths were used to incubate H. pluvialis for 39 days, and the growth rates were compared. As a result, the growth of H. pluvialis, a limited ingredient medium, produced the highest cell growth in the fluorescent light source while cell growth was the lowest in the blue+red LED. The growth of H. pluvialis in commercial medium Neo was highest in the fluorescent light source, and cell growth was lowest in the blue LED. In this study, the MBBM culture medium showed better results than the Neo culture medium. Microalgae grown in the fluorescent light source using the MBBM culture medium showed the best cell growth result in this study. The results were optimized for the culture medium, light source, and light quantity in H. pluvialis culture for the production of secondary metabolites and provide basic data for the mass culture of microalgae.

• Korean Journal of Animal Reproduction
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• v.17 no.4
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• pp.271-278
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• 1994

The aim of this study was to compare the two culture systems 1) co-culture with cumulus cells and 2) chemically defined medium supplemented with amino acids (CR1aa) and fetal calf serum (FCS) of in vitro produced bovine embryos from follicular oocytes in vitro. Bovine follicular oocytes were collected from ovaries of slaughtered cows and matured in TCM199 supplemented with 10% FCS and hormones (1$\mu\textrm{g}$/ml FSH-P and 1$\mu\textrm{g}$/ml oestradiol-17$\beta$)24 hours at 39$^{\circ}C$ under 5% CO2 in air. The capacitation of spermatozoa from ejaculated or frozen bull semen was induced by centrifugation through Percoll density gradient (45%, 90%). Then capacitated spermatozoa (1$\times$106/ml) were inseminated into 50${mu}ell$ droplet containing matured follicular oocytes and incubated for 40~42 hours. Cleaved embryos of 2~4cell stage were transferred to the co-culture with cumulus cells and/or CR1aa medium supplemented with FCS. In semen source, the developmental rates to the blastocyst and the hatched blastocyst stages were higher in ejaculated semen(27.6% and 14.9%) than those of frozen-thawed semen(18.3% and 11.8%), respectively. In two culture systems, the proportions of embryonic development upto the blastocysts and the hatched blastocysts were higher of CR1aa medium (22.1% and 12.1%) than those of cumulus cell co-culture (16.8% and 5.1%), respectively. The number of cells in exapnded blastocysts was slightly higher in cumulus cells co-culture (122.6$\pm$8.5) than that in CR1aa medium (117.9$\pm$5.9). The present results indicated that the early development of in vitro produced bovine embryos can be maintained efficiently in CR1aa medium as well as in co-culture with cumulus cells.