• Clinical and Experimental Reproductive Medicine
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• 제47권4호
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• pp.284-292
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• 2020

Objective: This study investigated whether adding outer-well medium to inhibit osmotic changes in culture media in a dry-type incubator improved the clinical outcomes of in vitro fertilization-embryo transfer (IVF-ET) cycles. Methods: In culture dishes, the osmotic changes in media (20 µL)-covered oil with or without outer-well medium (humid or dry culture conditions, respectively) were compared after 3 days of incubation in a dry-type incubator. One-step (Origio) and G1/G2 (Vitrolife) media were used. Results: The osmotic changes in the dry culture condition (308 mOsm) were higher than in the humid culture conditions (285-290 mOsm) after 3 days of incubation. In day 3 IVF-ET cycles, although the pregnancy rate did not significantly differ between the dry (46.2%) and humid culture (51.0%) groups, the rates of abortion and ongoing pregnancy were significantly better in the humid culture group (1.5% and 49.5%, respectively) than in the dry culture group (8.3% and 37.8%, respectively, p<0.05). In day 5 IVF-ET cycles, the abortion rate was significantly lower in the humid culture group (2.2%) than in the dry culture group (25.0%, p<0.01), but no statistically significant difference was observed in the rates of clinical and ongoing pregnancy between the dry (50.0% and 25.0%, respectively) and humid culture groups (59.5% and 57.3%, respectively) because of the small number of cycles. Conclusion: Hyperosmotic changes in media occurred in a dry-type incubator by evaporation, although the medium was covered with oil. These osmotic changes were efficiently inhibited by supplementation of outer-well medium, which resulted in improved pregnancy outcomes.

• Reproductive and Developmental Biology
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• 제36권3호
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• pp.141-145
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• 2012

The present study was conducted to develop a simple method for porcine oocyte maturation without $CO_2$ regulation. In experiment 1, we evaluated that the effect of $CO_2$ non-supplement on porcine oocyte maturation. Cumulusoocyte complexes (COCs) were collected from 2~6 mm follicles and divided into three groups (Control, tube-$CO_2$, and tube-non-$CO_2$). For control, COCs were cultured in 4-well multidish in a $CO_2$ incubator. For tube-$CO_2$, COCs were cultured in a round-bottom tube in a $CO_2$ incubator, and for tube-non-$CO_2$, COCs were cultured in a round-bottom tube sealed tightly without $CO_2$ supplement in a dry incubator. The proportion of oocytes reached to metaphase II (M-II) was not significantly different among three groups (87.9% to 91.4%). In experiment 2, we evaluated the effect of $CO_2$ non-supplement during oocyte maturation on development of embryos. Oocytes with a polar body were divided into two groups (Control and tube-non-$CO_2$) and applied 1.1 kV/cm or 1.2 kV/cm voltages for parthenogenetic activation. After activation, embryos were cultured for 6 days and examined the development. The proportion of embryos cleaved was not significantly different among treatment (86.3% to 91.5%). The proportion of embryo reached to blastocyst stage was not significantly different among treatment (13.9% to 25.2%). The cell number of blastocysts was not significantly different among treatment (29.0 to 32.4). In conclusion, oocytes cultured in a dry incubator without $CO_2$ supplement have enough competence to development after parthenogenetic activation. These results would be useful for transporting oocytes or embryos a long distance.

• 농업생명과학연구
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• 제53권2호
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• pp.15-26
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• 2019

In vivo 측정법은 비용이 많이 들고 연구기간이 길며 동물복지 측면에서 여러 문제점이 있다. 이에 대한 대안으로 지난 수십년간 in vitro 소화율 측정법이 개발되어 왔다. 본 총설에서는 대표적인 단위 동물로써 사양되고 있는 돼지의 소화기관 특징 및 소화작용에 대한 논하였다. 또한, 최근에 개발된 daisy II incubator 를 이용한 in vitro 방법을 통해 돼지에서 주요 곡류사료로 이용되고 있는 옥수수, 쌀, 소맥 및 대맥의 회장 및 전분 건물소화율을 측정하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제23권12호
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• pp.1587-1593
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• 2010

Pastures of the Apennines of Central Italy contribute to feed resources of high altitude (above 1,300 m sea level) grazing systems. The objective of this study was to evaluate the effectiveness of faecal extract from the yak (Bos grunnienes) as an alternative microbial inoculum to rumen fluid for estimation of digestibility of several forage species. Forage samples produced at high altitude were tested in this study: four legumes (Lathyrus sativus L., Lotus corniculatus L., Onobrychis viciaefolia L. and Trifolium pratense L.), three forbs (Achillea millefolium L., Potentilla reptans L. and Teucrium flavum L.) and one grass (Brachipodyum pinnatum L.) were incubated with yak rumen fluid or faecal extract. A large variability in chemical composition was observed among the species collected. Rumen liquor and faecal samples were collected from adult healthy yak. The $Daisy^{II}$ incubator was used to evaluate the nutrient digestibility of forages using rumen liquor as control and faecal extract as alternative microbial inoculum sources. Filter bags containing samples of browse species were added to the four digestion vessels along with their respective inoculum and then incubated for 48 h and dry matter (DM), organic matter (OM), crude protein (CP), neutral and detergent fiber (NDF) digestibility was determined. There was a significant relationship between estimates, indicating that faecal liquor has the potential to be used instead of rumen fluid for estimation of in vitro digestibility of plants. It is concluded that the $Daisy^{II}$ incubator results are appropriate for the determination of in vitro digestibility of nutrients using faecal liquor to define the potential for adaptation of yak to new pastures.

• 한국작물학회지
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• 제34권2호
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• pp.147-154
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• 1989

일본형 수도, 통일형 수도 및 통일형 수도의 교배친 등을 공시하여 이의 종근으로부터 근단을 1cm로 절취한 분리근을 천전 등의 $\ulcorner$개량배지$\lrcorner$를 기본으로 하여 pH, 당농도, Casamino acid 농도 등을 달리한 oq지에 무균배양한 분리근의 생장 튿성을 비교하였던 바 얻어진 결과를 요약하면 다음과 같다. 1. 분리근의 근장은 27$^{\circ}C$구에서는 일본형품종과 다수형 품종간에 별차이는 없었으나 3$0^{\circ}C$구에서는 일본형 수도에 비교해서 통일형 품종의 신장이 현저히 양호하였다. 2. 분리근의 건물중은 27$^{\circ}C$구 및 3$0^{\circ}C$구 공히 일본형 품종에 비교해서 통일형 품종이 현저하게 무거운 것으로 인정되었다. 3. 일본형 품종과 통일형 품종간에 나타난 분리근의 건물중 차이는 주로 측근발육의 양부에 따른 것으로 특히 2~3cm길이의 특근수에 차이가 인정되었다. 4. Casamino acid 반응에서 일본형 품종은 주근 및 측근의 생장이 0.2%구에서 양호하였으나, 통일형 품종에서는 대부분의 품종이 주근의 신장은 Casamino acid 무처리구에서 측근의 생장은 0.2%구에서 매우 양호하였다. 5. 분리근의 생장은 배지의 pH, 당농도 및 Casamino acid 농도 등을 달리한 경우에 있어서도 통일형 품종이 일본형 품종보다 양호하였다. 6. 통일형 품종의 교배친중 인도별 품종과 일본형 품종에 있어서 전자는 후자보다 분리근의 생장이 양호하였을 뿐만 아니라 Casamino acid반응이 통일형 품종과 거의 같은 경향인 것으로 나타났다.

• 농업과학연구
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• 제29권2호
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• pp.20-34
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• 2002

본 연구는 Aspergillus oryzae 배양물이 in vitro 건물, 섬유소 및 단백질 소화율과 pH에 미치는 영향을 구명하고자 실시하였다. 균종 Aspergillus oryzae는 한국 생명공학연구원에서 분양받았으며, 본 연구에 사용된 사료는 시중 유통중인 착유우용 배합사료와 TMR 사료로서, 일반성분 분석은 (주)우성사료 중앙실험실에서 실시하였고, 건물, 섬유소 및 단백질 소화율 실험은 충남대학교 실험실에서 실시하였다. 배합사료와 TMR에 Aspergillus oryzae 배양물 0, 1.0, 2.0 및 3.0%를 첨가한 후, Holstein종 착유우로부터 채취한 반추위액에 시료 2.0g 내외를 넣어 진탕배양기에서 24, 48 및 72시간 소화시켰다. 침전물에 0.2% pepsin이 들어있는 0.1N HCl 30ml를 넣고 $39^{\circ}C$ incubator에서 48시간 소화시키고 나서, 마지막으로 침전물을 $60^{\circ}C$의 건조기에서 48시간 건조시켰다. 실험은 3회 반복하여 실시하였다. 건물 소화율은 Aspergillus oryzae 발효물을 첨가하지 않은 대조구에 비해 24시간 배양시킨 배합사료의 경우 2.1%(63.1%), 9.7%(68.5%) 및 9.0%(68.0%) 개선되었고, TMR은 4.8%(60.0%), 6.4%(61.1%) 및 2.9%(58.8%)의 개선효과가 있었다. 이와 대조적으로, 48시간 및 72시간 배양시킨 시험구의 건물 소화율에 Aspergillus oryzae 배양물이 미치는 영향은 24시간 배양시보다 상대적으로 낮은 경향을 보였다. 섬유소 소화율에서 Aspergillus oryzae가 조섬유, ADF 및 NDF 소화율을 유의적으로 개선시킨다는 것을 알 수 있었다. 배합사료에서 3% 첨가구의 경우 24시간 배양시 13.3%(53.3%), 72시간 배양시 2.4%(54.6%)까지 증가되었다. 하지만, 첨가수준에 따라 소화율이 높아졌음에도 불구하고 첨가수준별 개선효과에는 유의적인 차이가 없었다. 단백질 소화율은 Aspergillus oryzae 배양물 첨가에 의해 배합사료에서 0.4%(79.7%)에서 9.4%(71.8%)의 유의적인 개선효과를 나타내었지만, 2.0%와 3.0% 첨가구간의 유의적인 차이는 인정되지 않았다. TMR의 경우에는 4.0%(70.4%)에서 6.3%(65.1%)의 유의적인 소화율 향상을 나타내었지만 2.0%와 3.0% 첨가구간의 유의적인 차이는 인정되지 않았다. 본 연구에서, pH 처리구간 유의적인 차이는 없었으나, Aspergillus oryzae 배양물 첨가에 따라 pH가 약간 감소하는 경향을 보였다. 그러므로 본 발효물은 pH에는 영향을 미치지 않으나 건물, 섬유소 및 단백질 소화율 향상에는 다소 효과가 있 는 것으로 결론을 내릴 수가 있다.

• KSBB Journal
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• 제11권4호
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• pp.398-404
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• 1996

전세포 효소 고정화 캡슐을 제조하기 위하여 Eschericia coli를 calcium alginate 캡슐내부에 접종하고 배양하였다. E. coli의 캡슐내부 건조중량이 1OOg/L에 달하였다. 생산배지에서 배양하는 동안 캡슐내부에 축적되는 미생물의 양이 많을수록 캡슐 내부에 축적되는 $\beta$galactosidase의 활성도 높았다. 생산배지에 금속이온 $Zn^{+2}를 2{\times}10^{-4} M $ 첨가함으로써 캡슐내부에 축척되는 ${\beta}$-galactosidase의 활성 을 25% 증가시킬 수 있었다. 캡슐제조시 해바라기유를 부피비로 2% 첨가함으로써 캡슐내부에 축적되 는 ${\beta}$-galactosidase의 활성을 10% 증가시킬 수 었었다. 부피산소전달계수, $k_La가 2.55h^- $ 안 플라스크 대신 $k_La가 82h^- $인 concentric air lift reactor 내에서 고정화 E. coli를 배양호농로써 캡슐내부의 전세포${\beta}$-galactosidase의 활성을 86% 증가시킬 수 있었다.

• 한국약용작물학회지
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• 제20권6호
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• pp.479-486
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• 2012

The production of adventitious roots derived from root explant of Echinacea angustifolia and its secondary metabolite content were assessed in different types and levels of auxin. The induction of adventitious roots from root explant cultured in Murashige and Skoog solid medium supplemented with 1.0 mg/L indole -3-butyric acid (IBA) attained highest as 20.87 mg fresh weight and 3.07 mg dry weight per culture but root suspension culture at the same concentration of IBA enhanced biomass production as 3.07 g fresh weight and 0.38 g per culture after 4 weeks in culture. 3.0 mg/L ${\alpha}$-naphthalene acetic acid (NAA) treatment had similar effect on root biomass production as 3.07 g fresh weight and 0.38 g per culture with liquid suspension culture, whereas adventitious roots exposed to over 3.0-5.0 mg/L IBA or 5.0 mg/L NAA were less responsive by reducing the number of adventitious roots and/or changing root morphology such as short and thick. The content of secondary metabolites such as phenolic, flavonoids and total caffeic acid in adventitious roots cultured on MS medium supplemented with 1.0 mg/L IBA were attained highest as 27.20, 9.60. 10.67 mg/g dry weight, respectively. Overall, the best production of root biomass and secondary metabolites were given by 1.0 mg/L IBA.

• Asian-Australasian Journal of Animal Sciences
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• 제28권2호
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• pp.188-199
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• 2015

Nineteen tanniferous browse plants were collected from South Africa to investigate their digestibility, gas production (GP) characteristics and methane production. Fresh samples were collected, dried in forced oven, and ground and analyzed for nutrient composition. In vitro GP and in vitro organic matter digestibility (IVOMD) were determined using rumen fluid collected, strained and anaerobically prepared. A semi-automated system was used to measure GP by incubating the sample in a shaking incubator at $39^{\circ}C$. There was significant (p<0.05) variation in chemical composition of studied browses. Crude protein (CP) content of the species ranged from 86.9 to 305.0 g/kg dry matter (DM). The neutral detergent fiber (NDF) ranged from 292.8 to 517.5 g/kg DM while acid detergent fiber (ADF) ranged from 273.3 to 495.1 g/kg DM. The ash, ether extract, non-fibrous carbohydrate, neutral detergent insoluble nitrogen, and acid detergent insoluble nitrogen and CP were negatively correlated with methane production. Methane production was positively correlated with NDF, ADF, cellulose and hemi-cellulose. Tannin decreased GP, IVOMD, total volatile fatty acid and methane production. The observed low methanogenic potential and substantial ammonia generation of some of the browses might be potentially useful as rumen manipulating agents. However, a systematic evaluation is needed to determine optimum levels of supplementation in a mixed diet in order to attain a maximal depressing effect on enteric $CH_4$ production with a minimal detrimental effect on rumen fermentation of poor quality roughage based diet.

• Journal of Microbiology and Biotechnology
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• 제28권11호
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• pp.1865-1875
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• 2018

Enhanced application of solid-state fermentation (SSF) in industrial production and the influence of SSF of Rhizopus K1 on glucoamylase productivity were analyzed using the flat band method. A growth model was implemented through SSF of Rhizopus K1 in this experiment, and spectrophotometric method was used to determine glucoamylase activity. Results showed that in bran and potato culture medium with 70% moisture in a loose state, ${\mu}$ of mycelium reached to $0.15h^{-1}$ after 45 h of culture in a thermostatic water bath incubator at $30^{\circ}C$. Under a low-magnification microscope, mycelial cells appeared uniform, bulky with numerous branches, and were not easily ruptured. The generated glucoamylase activity reached to 55 U/g (dry basis). This study has good utilization value for glucoamylase production by Rhizopus in SSF.