Recent progress has been made to establish intestinal organoids for an in vitro model as a potential alternative to an in vivo system in animals. We previously reported a reliable method for the isolation of intestinal crypts from the small intestine and robust three-dimensional (3D) expansion of intestinal organoids (basal-out) in adult bovines. The present study aimed to establish next-generation intestinal organoids for practical applications in disease modeling-based host-pathogen interactions and feed efficiency measurements. In this study, we developed a rapid and convenient method for the efficient generation of intestinal organoids through the modulation of the Wnt signaling pathway and continuous apical-out intestinal organoids. Remarkably, the intestinal epithelium only takes 3-4 days to undergo CHIR (1 µM) treatment as a Wnt activator, which is much shorter than that required for spontaneous differentiation (7 days). Subsequently, we successfully established an apical-out bovine intestinal organoid culture system through suspension culture without Matrigel matrix, indicating an apical-out membrane on the surface. Collectively, these results demonstrate the efficient generation and next-generation of bovine intestinal organoids and will facilitate their potential use for various purposes, such as disease modeling, in the field of animal biotechnology.

Methyl Bromide (MB) has been regulated by montreal protocol due to ozone depletion since 1992, and there is a need to develop MB alternative fumigation methods. To find out the efficacies and tolerance in developmental stages of citrus mealybugs (Planococcus citri) which frequently infested in imported pineapples, phosphine(PH3) gas was tested in different exposure times (2~24 hr) and concentrations (0.1∼2g/m3) in small desiccators at 8℃. Based on Lethal 99% concentration × time products (LCTP 99%) of PH3 gas to target developmental insect, 2g/m3 PH3 was applied for 24 hr in 300 m3 fumigation facility, which was designed in well sealed and evacuation system (5 air exchange/min). We monitored PH3 gas in fumigation facility to confirm CTP and checked PH3 gas in atmospheres in air ventiliation process after fumigations to make quarantine guideline and protocol for worker safety as well as efficacy to target pest.

Background : This study, the fraction for testing the efficacy of the Astragalus extract was concentrated active ingredient. The concentrated fraction was applied to a cosmetic material that Astragalus testing results confirmed that the improved efficacy. Methods and Results : The fractions were performed using an n-butanol solvent for increasing the efficacy of the Astragalus extract, by using the material fractions collected to compare and ultimately an increase in whitening and wrinkle efficacy. The solvent to be used in the fractions was used for the n-butanol good dissolution to the effective substance(Astragaloside, Isoflavonoid). It increased approximately 6.5 times the sample extract and the n-butanol fraction of the Astragalus as a result Astragaloside 15 ppm, 97 ppm respectively analyzed by HPLC equipment, isoflavonoid content was confirmed by an increase in the content of the fractions increased 4.5 times to 280 ppm, 1,260 ppm. Tyrosinase inhibitory effect, respectively IC50 5.70 mg/mL, IC50 1.02 mg/mL to, Collagenase producing ability is IC50 4.88 mg/mL, IC50 0.93 mg/mL with n-butanol fraction was good whitening, anti-wrinkle efficacy than the extract. Sensory evaluation was conducted in the same amount of sample, using a purified Astragalus cosmetics received high marks. Stability evaluation(MTT assay) was checked for more than 100% cell viability at the concentration 2,000 ppm. Conclusion : n-butanol fraction of Astragalus was subjected to a component analysis and In vitro test, it was confirmed an increase active ingredient content. The results of sensory evaluation and stability evaluation, it was confirmed been made to improve qualities as a cosmetic materials.