Emerging RNA viruses continue to pose serious threats to animal and human health, necessitating the development of broad-spectrum antiviral agents within a One Health framework. In this study, we evaluated the antiviral potential of Desmodesmus multivariabilis extracts against three representative RNA viruses: influenza A virus (IAV), porcine epidemic diarrhea virus (PEDV), and hepatitis A virus (HAV). Extracts derived from four closely related microalgal isolates were tested using pre- and post-treatment protocols in vitro. Among them, strain ME749 demonstrated the most potent and consistent antiviral activity, with low cytotoxicity and high selectivity index (SI), particularly against IAV (SI = 23.8), followed by PEDV (SI = 10.7) and HAV (SI = 6.2). ME752 also showed moderate antiviral efficacy. RT-qPCR confirmed significant reductions in viral RNA levels for all three viruses, with ME749 achieving up to 3-log₁₀ reduction in IAV and 2.4-log₁₀ reduction in HAV. These results suggest that the antiviral effect may be mediated through mechanisms beyond direct virucidal activity, potentially involving host immune modulation. Although the active compounds remain unidentified, carotenoids are hypothesized as key bioactive components. This study highlights Desmodesmus multivariabilis, particularly ME749, as a promising source of novel, natural antiviral agents with applications in both veterinary and zoonotic disease control. Future studies including metabolomic profiling, mechanism-of-action analyses, and in vivo validation are warranted to further explore its potential in combating current and emerging RNA virus threats.

Background: Embryo implantation is a complex process regulated by interactions between endometrial epithelial and stromal cells. The endometrium plays a critical role in this process, providing a supportive environment for embryo attachment. However, conventional 2D cell culture models fail to fully replicate the complex 3D structure and cellular interactions of the endometrium. To overcome these limitations, 3D organoid models have been developed to better mimic the in vivo endometrial environment. Methods: In this study, a multicellular uterine organoid model was developed using porcine endometrial epithelial cells (pEECs) and porcine endometrial stromal cells (pESCs) to evaluate the effects of the endometrial environment on embryo implantation. First, single-cell endometrial organoids (pEOs) were formed by culturing pEECs in Matrigel, and their basic cellular characteristics were assessed. Then, a multicellular uterine organoid model was established by combining pEOs with pESCs. Finally, porcine embryos were co-cultured with this model to examine its effect on embryo attachment. Results: The multicellular uterine organoid model facilitated embryo attachment, demonstrating that the 3D structure and cellular interactions of the endometrium play a significant role in embryo implantation. The presence of both epithelial and stromal cells contributed to a more physiologically relevant environment that supported embryo adhesion. Conclusions: This study demonstrates that a multicellular uterine organoid model can serve as a useful in vitro system for porcine embryo implantation research. This model may contribute to a better understanding of embryo development and implantation mechanisms, with potential applications in regenerative medicine and biotechnology.

본 연구는 발효 황련해독탕(FHHT)의 식후혈당 상승 억 제 효능을 검증하기 위해 농도별로 Rat 유래 α-glucosidase 와 돼지 췌장 유래 α-amylase에 대한 저해 활성과 sprague dawley rat 모델을 이용한 식후혈당 상승 억제 효능을 평가 하였다. 그 결과, FHHT는 농도 의존적으로 α-glucosidase와 α-amylase의 소화 효소 활성을 저해하였고 IC₅₀ 값은 α- amylase 9.321mg/mL, α-glucosidase 0.143mg/mL, maltase 1.159mg/mL, glucoamylase 0.263mg/mL, sucrase 0.112mg/ mL로 확인되었다. 동물 모델 실험에서는 FHHT가 다당류 starch 및 이당류 sucrose 섭취 후 통계적으로 유의한 식후 혈당 상승 억제 효능을 보였다(P<0.01). 이와 같은 결과를 통해 FHHT의 당 흡수 억제를 통한 항당뇨 효능을 확인 하였고 추가 연구의 필요성 및 시사점을 제시함으로써 본 연구가 항당뇨의 새로운 한방 건강 기능식품으로 활용되 기 위한 참고 자료가 될 것으로 사료된다.

포도 ‘My heart’의 기내증식과 기외이식 후 생장에 있어서 배양용기에 부착한 미세공극 Filter 처리가 건전한 유묘를 생 산하는데 효과적인 방법을 찾고자 실시하였다. 미세공극 Filter는 환기구 크기별로 White filter type (50.0 mm×3.5 mm)과 Green filter type (50.0 mm×7.0 mm) 으로 구분하여 밀폐 처리와 비교하였다. Shoot tip 배양에 있어서 Green filter type에서 shoot 분화율이 75%로 White filter type 77% 와 밀폐처리 80% 보다 낮았지만 투명화 shoot 발생율은 4% 로 White filter type 13.4%, 밀폐처리 24.7%에 비하여 9.4－ 20.7%가 적었다. 전체 배양묘의 54.9%가 투명화 발생이 되 었을 때 Green filter type으로 90일 동안 계대 배양하고 조사 한 결과 투명화율은 11.8%로 감소하였고 분화한 shoot 수는 89개에서 915개로 증가하였다. Filter type에 따라 IBA 2.0 mg·L-1를 첨가하여 30일 동안 배양을 하였을 때 Green filter type에서 기내 발근율 100%, 뿌리 수 7.3개, 엽수 10.0개로 White filter type과 밀폐 처리보다 좋았다. 기외 이식하고 15 일 후의 유묘 생존율도 Green filter type에서 100%로 다른 처 리에 비하여 1.5－29.5% 더 높았고 초장이 11.0cm, 생체중이 1.7g 으로 가장 양호하였다. 미세공극 Green filter 처리는 포 도 ‘My heart’의 기내배양에서 shoot 투명화를 감소시키고 shoot와 뿌리 생성을 촉진시키고 기외이식 후 생장은 통계적 으로 유의하게 확인되어 건전한 유묘 생산에 효과적이었다.

This study investigated the growth dynamics of Lilium miquelianum bulb scales cultured in four different media formulations: Full Murashige-Skoog (FMS), Half Murashige-Skoog (HMS), Anderson Vitamin (AV), and Knudson Vitamin (KV). Bulb scales were collected from native populations on Jeju Island, Korea, and cultured in vitro for 12 weeks. Growth parameters, including bulb area and scale count, were measured at 3-week intervals. Statistical analyses revealed significant differences in growth rates across media types and time points. FMS consistently demonstrated the highest growth rates and final bulb area and scale count, while KV exhibited the lowest performance. Heatmap analysis showed that FMS achieved the highest weekly growth rates in most time points, with HMS showing comparable performance during early growth stages. Individual sample growth rates varied within media, highlighting the influence of both external and internal factors on growth. Correlation analysis revealed a positive association between bulb size and scale quantity, with FMS exhibiting the strongest relationship. These findings emphasize the importance of appropriate media selection for optimal L. miquelianum propagation, with FMS being the most suitable for extended cultivation. This research provides valuable insights for improving propagation efficiency and conservation efforts of L. miquelianum.

Habenaria radiata, commonly known as the Egret Flower, is a critically endangered terrestrial orchid facing rapid population decline due to overharvesting in Korea. Asymbiotic seed propagation offers a promising conservation method for this rare species. This study investigates the effects of different nutrient media on the germination and plantlet development of immature seeds of H. radiata, with the goal of establishing optimal propagation conditions. Among four media types, with the highest germination rate (75.6%) observed on OBTSG medium, followed by 1/2 MS, OMM, and OSGM. Protocorm development varied across media, with OBTSG and OMM supporting the most robust growth. A two-step culture method involving OBTSG for germination followed by OMM for plantlet development proved most effective, achieving an 84.4% survival rate. These findings highlight the importance of media composition and transitioning for successful propagation. The results provide a practical protocol for in vitro propagation, contributing to both in situ and ex situ conservation efforts aimed at preserving H. radiata and other endangered terrestrial orchids.

Background: This study was conducted to increase meat production by 30% compared to the present by selecting a giant cow over 1,000 kg and applying biotechnologies. Methods: After OPU from 1,100 kg of giant cow, the calves were produced in Hanwoo surrogate mothers. Among 23 calves six male heads were selected, three heads of them were chosen as candidate sires, and the rest three heads were raised for 30 months for performance test. The semen of three candidate sires from the age of 22 months were collected and frozen, and the calves were produced by artificial insemination. The calf was raised to the age of 30 to 33 months and performed a progeny test. Results: The average birth weight of 23 calves born by transferring giant cow-derived embryos was 42.8 kg, and the average weight of carcass from three bulls was 615.3 kg in the performance test. In the progeny test, the average birth weight of calves born after artificial insemination of semen from giant cow-derived candidate sires was 41 kg, and the average weight of carcass after raised to the age of 30 to 33 months was 562.7 kg. As a result of performance and progeny tests, it is 148 and 96 kg higher in giant cow-derived beef cattle than the average carcass (467 kg) from general Hanwoo cattle, respectively. Conclusions: This study will have laid a great foundation for the future improvement of the Korean beef industry.

Fetal Bovine Serum (FBS) plays a crucial role in animal cell culture; however, the increasing number of bovine fetuses used and sacrificed solely for FBS collection has raised ethical concerns globally. The welfare of fetuses during FBS blood collection has become a key focus of debate among animal welfare and ethics organizations worldwide. Previous studies indicate that heat-inactivated coelomic fluid (HI-CF) from the earthworm Perionyx excavatus may serve as a viable FBS alternative in adherent cell cultures. This study evaluates the potential of HI-CF as an FBS substitute during the in vitro maturation (IVM) stage of bovine embryo culture, with a focus on improving developmental rate through antioxidation effects. In this study, 2% HI-CF was incorporated into IVM media, assessing its impact on cell growth, differentiation, and the expression of genes related to antioxidation. The group of 2% of HI-CF exhibited a trend toward increased cleavage and blastocyst development rates compared to the control group. Although antioxidant genes such as NRF2 and GSR showed no statistically significant differences between the control and treatment groups, a trend toward increased expression was observed. Conversely, GPX1 displayed a trend of decreased expression. Notably, IGF1 and NQO1 were significant upregulated (p < 0.05) in the 2% HI-CF group. Additionally, oocytes stained with H2DCFDA showed a significantly reduced ROS levels (p < 0.05) in the 2% HI-CF group compared with controls. These findings suggest that HI-CF's antioxidative effects support enhanced cell growth and blastocyst development rate, surpassing those observed with FBS. Consequently, HI-CF shows promise as an effective alternative to FBS in vitro maturation of bovine oocytes.

The objective of the present study was to investigate the effects of different red seaweeds on in vitro rumen fermentation characteristics and methane gas production. Five species of red seaweed (Chrysymenia wrughtii Yamada, CW; Hypnea sp., Hypnea sp.; Chondria crassicaulis, CC; Gelidium vagum Okamurae, GV; Hypnea saidana Holmes, HS) were obtained from National Institute of Fisheries Science (NIFS) in South Korea. The collected red seaweeds were washed for 3 minutes, and then samples were freeze-dried and ground to a size of a 1 millimeter. The buffered ruminal fluid (50 mL) was incubated with substrates and seaweeds (5% of substrates) at 39℃ for 48 hours. Total gas production was lower than red seaweed treatments excluding the CW treatment (p<0.05; 63.25 mL). Methane production was the lowest in CC treatment (p<0.05; 9.93 mL/g of digestible dry matter). The rumen pH of the red seaweed treatments ranged from 5.98 to 6.08, which was significantly the lowest in the GV treatment (p<0.05; 5.98). There was no significant difference in the total VFA concentration, but propionate (27.53%) was significantly highest in the CW treatment, whereas acetate (53.14%), iso-valerate (3.52%), valerate (1.72%), and A:P ratio (1.93) were significantly lowest (p<0.05). In conclusion, among the five species of red seaweeds, Chondria crassicaulis reduced in vitro methane production without negative effects on dry matter digestibility. Future studies will be needed to determine the optimal inclusion level of Chondria crassicaulis as feed additive to reduce enteric methane production.

This study aimed to investigate the effects of various washing pre-treatments of native Codium fragile as a feed additive on in vitro ruminal fermentation and CH4 production in ruminants. Seaweed was included at 0.5% dry matter (DM) based on the experimental feed (forage : concentrate = 3:7). Treatment groups were classified as follows: experimental feed (C), no washing (T1), washing at 0°C (T2), washing at 22°C (T3) and washing at 70°C (T4) each immersed for 6 minutes in distilled water. The pH consistently fell within the ruminal stability range. In vitro dry matter digestibility was significantly highest in T2, T3, T4 and C, T4 was the lowest at 48 h (p<0.05). NH3-N concentration was significantly highest in T4 at 48 h (p<0.05). Total gas production at 48 h was 19% lower in T4 compared to C (p<0.01). CH4 production (mL/g DM) at 48 h was lower in all treatment groups compared to C, with T3 showing a 31% reduction (p<0.01). Similarly, CH4 production (mL/g dry matter degradability, DMD) at 48 h was 39% lower for T3 compared to C (p<0.01). At 24 h, total VFA was significantly highest in T1 and T4 (p<0.05). The proportions of acetate was significantly highest in C and T3 was the lowest at 48 h (p<0.01). The proportions of propionate was significantly highest in T3 and C was the lowest at 48 h (p<0.01). The acetate to propionate ratio was singnificantly highest in C at 48 h (p<0.01). The proportions of butyrate at 24 h was lower for T3 compared to C (p<0.05). Therefore, this study confirms that Codium fragile can reduce CH4 production when used as a feed additive for ruminants and this effect is not significantly influenced by the washing pre-treatment. However, if washing process is necessary, washing at 22°C is the most appropriate method to remove foreign objects.

Drug-induced liver injury (DILI) is considered to be a significant cause of drug wastage. To mitigate clinical DILI risks, assessing drugs using human liver models is crucial since animal studies may fall short due to species-specific liver pathway variations. Cell-based preclinical hepatotoxicity testing is often pertinent. In the present study, cells from a human liver cancer line (HepG2 and HepaRG) were cultured in both formats of 2D and 3D spheroids to explore their responses to drugs. Liver-specific marker expressions across cell lines and culture formats were also examined to assess disparities in DILI marker expressions. After treating each cell with the drugs, cytotoxicity and liver injury markers aspartate aminotransferase and alanine aminotransferase were increased. In addition, liver specific markers albumin and urea decreased in a drug concentration-dependent manner. These findings were consistent with drug sensitivity. Additionally, mRNA expression levels of cytochrome P450 enzymes (CYPs) involved in hepatocellular drug metabolism were compared following treatment with enzyme inducers. CYP1A2 and CYP2C9 were not epxressed in HepG2 cells. HepaRG cells exhibited significantly increased expression of CYP1A2, 2C9, and 3A4 post-treatment. Notably, enzyme expression was notably higher in 3D cultures than in 2D cultures. Collectively, these findings suggest that HepaRG cells and 3D cultures hold promise for evaluating DILI during early-stage drug development.

The study aimed to investigate the impact of varying levels of neutral detergent-soluble fiber (NDSF) in Hanwoo growing cattle diets on rumen fermentation and methane (CH4) emissions. An in vitro rumen fermentation experiment utilized feeds with different NDSF levels, incorporating ingredients such as corn grain, soybean meal, soybean hulls, palm kernel meal, beet pulp and timothy hay. The NDSF contents in the diets were 9.02% (T1), 10.09% (T2), 12.42% (T3) and 14.63% (T4). In vitro dry matter digestibility (IVDMD) at 48 h was 7.4% higher for T4 compared to T1 (p<0.05). Total gas production at 48 h was 6.6% higher for T4 than T1 (p<0.05). CH4 production significantly decreased at 9 h and 12 h for T1 and T2 (p<0.05). At 48 h, CH4 production was 5.6% higher for T4 compared to T1 and 6.7% higher compared to T2 (p<0.05). At 12 h ans 24 h, the ammonia nitrogen concentration of T4 was approximately 33.1% and 40.4% lower, respectively, compared to T1 (p<0.05). The acetate to propionate ratio at 48 h was approximately 18.8% higher for T4 than T1 (p<0.05). From 9 h to 48 h, the proportions of butyrate and valerate were significantly higher for T4 (p<0.05). At 48 h, the dominant phylum in T4's rumen microbial community was Candidatus Thermoplasmatota Methanomassiliicoccus, an Archaea. Therefore, this study confirmed that increasing the NDSF content in growing Hanwoo cattle diets up to 12.42% increases IVDMD without increasing CH4 emissions, which is expected to positively impact Hanwoo productivity.

This study was conducted to estimate the effects of the forage process on rumen fermentation characteristics and greenhouse gas emissions of rye. Rye was grown at the Taeyoung Livestock farm and harvested at the heading stage. The harvested rye (5 kg) was sub-sampled for fresh forage, hay, and silage in triplicates. The sub-sampled rye was freeze-dried or air-dried for fresh forage or rye hay, respectively. For rye silage, the sub-sampled rye forage was ensiled into a 10 L mini bucket silo and stored for 90 days. For 72 h rumen incubation, each forage (0.3 g) was placed into the incubation bottle with the rumen mixture (30 mL) in quadruplicates. After the incubation, total gas was measured and sub-sampled for CO2 and CH4 analyses, and the bottle content was centrifuged for in vitro digestibilities of dry matter (IVDMD) and neutral detergent fiber (IVNDFD), and rumen fermentation characteristics. Silage had higher crude protein, crude ash, and acid detergent fiber concentrations than fresh forage and hay but lower non-fiber carbohydrates and relative feed value (p<0.05). And, silage had higher lactic acid bacteria than the other forages but lower pH (p<0.05). After 72 h incubation in the rumen, fresh forage had higher IVDMD and butyrate content than the other forages (p<0.05). However, silage had higher rumen pH and propionate content than the other forages but lower A:P ratio (p<0.05). Regarding greenhouse gases, silage had lowest total gas (mL/g DMD and NDFD) and CH4 (mL/g DMD and NDFD) emissions, while fresh forage had lowest CO2 (mL/g DMD) emission (p<0.05). Therefore, this study concluded that the ensiling process of rye can effectively mitigate greenhouse gas emissions of Hanwoo.