• Clinical and Experimental Reproductive Medicine
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• v.47 no.4
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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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• v.36 no.3
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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.

• Journal of agriculture & life science
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• v.53 no.2
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• pp.15-26
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• 2019

An in vitro digestibility methods have been developed to mimic an in vivo system in the past decades. Because the in vivo techniques cause high cost, intensive labor, longer research periods and ethnical problems. In this review, the digestive systems from stomach to large intestine of pig as a monogastric animal were addressed to understand an in vivo digestion. The innovative in vitro technique using the Daisy II incubator was performed and the in vitro ileal and fecal dry matter (DM) digestibility of corn, rice, wheat and barley was determined.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.12
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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.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.34 no.2
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• pp.147-154
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• 1989

The experiments were carried out to compare the growth characteristics of excised seminal roots in Japonica type, Indica- Japonica hybrids and its parents. Based on the modified culture medium by Kawata et al., several levels of pH. sucrose and casamino acid were varied in the culture media. The excised root length at 27$^{\circ}C$ incubator was little different between Japonica type and Indica-Japonica hybrids, but at 30$^{\circ}C$ incubator the excised root elongation of Indica-Japonica hybrids was much better than that of Japonica type. The dry matter weight of excised root in Indica- Japonica hybrids was significantly heavier than in Japonica type at both 27 and 30$^{\circ}C$ incubators. The difference of dry root weight between Japonica type and Indica- Japonica hybrids was caused from the number of lateral roots, especially the root numbers of 25 cm length. The main and lateral seminal root growth of Japonica type grew best at the lot of 0.2 percent casamino acid. However, the lots of 0 and 0.2 percent casamino acid showed good results in the main and lateral excised root growth. respectively of Indica-Japonica hydrids. The growth of excised roots was better in Indica-Japonica hybrids than in Japonica type at even various levels of pH, sucrose and casamino acid. Between the parents of Indica - Japonica hybrids, the excised root growth of Indica type was more predominant than that of Japonica type, and the effect of casamino acid on the excised root growth in the formar was similar to that of Indica type. This result indicated that the growth characteristics of excised seminal roots in Indica Japonica hybrids were derived from the parent, Indica type.

• Korean Journal of Agricultural Science
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• v.29 no.2
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• pp.20-34
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• 2002

This study was conducted to examine the effects of an Aspergillus oryzae fermentation culture on the in vitro digestibilities of dry matter, crude fiber, acid detergent fiber (ADF), neutral detergent fiber (NDF), crude protein, and pH in in vitro experiment of diets for dairy cows. A fungal species, Aspergillus oryzae was supplied by Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea (KCTC 1229). The experimental diets were commercial compound feed (concentrate) and total mixed ration (TMR) for lactating cows, of which chemical analyses were determined at Research and Development Institute, Woosung Feed Co., Ltd., while the digestibilities were done at the laboratory of Chungnam National University. Aspergillus oryzae culture products were added to compound feed and TMR at the rate of 0, 1.0, 2.0, 3.0% respectively. The experimental diet with the rumen fluid sampled from Holstein fresian milking cows were used and digested for 24 hrs, 48hrs and 72hrs in the shaking incubator. The residues of the digesta were digested for 48hrs in the incubator in which put 30ml of 0.1N HCl with 0.2% pepsin at $39^{\circ}C$. The final precipitates were dried for 48hrs in the drier at $60^{\circ}C$. These experimental procedures were triplicated to determine the in vitro digestibility of dry matter, crude fiber, ADF, NDF, crude protein and pH. Compared to control diet, not added Aspergillus oryzae, the DM digestibility of fungal diets were improved 2.1%(63.1%), 9.7%(68.5%) and 9.0%(68.0%) for 24 hour fermentation in compound feed while 4.8%(60.0%), 6.4%(61.1%) and 2.9%(58.8%) in TMR. On the contrary, for 48 hour and 72 hour digestibilities, the effects of Aspergillus oryzae culture on the digestibility of dry matter were relatively lowered compared to 24 hour digestibility. Referring to the digestibility of dietary fiber, Aspergillus oryzae was believed to significantly improve digestibilities of crude fiber, ADF and NDF. Those were increased up to 13.3%(53.3%) for 24 hour fermentation, while 2.4%(54.6%) for 3.0% added for 72 hour fermentation in compound feed. However, there were no significant differences among the treatments for the inclusion rate of Aspergillus oryzae, even though the more inclusion rate, the better digestibility. The protein digestibilities were significantly improved from 0.4%(79.7%) to 9.4%(71.8%) by adding Aspergillus oryzae into compound feed. However, there were no significant differences between the two experimental diets, 2.0% and 3.0% Aspergillus oryzae included diets. In case of TMR, the protein digestibilities were significantly improved from 4.0%(70.4%) to 6.3%(65.1%) by adding Aspergillus oryzae. However, there were no significant differences between the two experimental diets, 2.0% and 3.0% Aspergillus oryzae included diets. In this study, there were no significant differences among the treatments in pH. On the contrary, there were slightly decrease in pH by adding Aspergillus oryzae into experimental diets but not significant. Summarizing the results of this examination, Aspergillus oryzae fermentation culture is believed to improve the digestibilities of dry matter, fiber and crude protein in cattle diets. However, more detailed research for the mechanism of the fungal culture is required to improve ruminal environment.

• KSBB Journal
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• v.11 no.4
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• pp.398-404
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• 1996

Escherichia coli was inoculated in calcium alginate capsules and cultivated to prepare encapsulated whole cell ${\beta}$-galactosidase. The dry cell weight in the capsule reached 100 g/L based on the inner space of the capsule. The activity of the encapsulated whole cell ${\beta}$-galactosidase increased with the dry cell weight increase during cultivation in the production medium. The activity of the encapsulated whole cell ${\beta}$-galactosidase was increased 25% by adding $2{\times}10^{-4}M Zn^{+2}$ ion in the production medium and 10% by coencapsulating with 2% (v/v) sunflower seed oil. The activity of encapsulated whole cell ${\beta}$-galactosidase produced in the concentric air lift reactor in which kLa was 82/hr was 86% higher than that in the shaking flask incubator where kLa was 2.55/hr.

• Korean Journal of Medicinal Crop Science
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• v.20 no.6
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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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• v.28 no.2
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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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• v.28 no.11
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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.