Probiotics have been evaluated as therapeutic agents for cancer treatment in an increasing number of studies. This study investigated the inhibitory and cytotoxic effects of specific Lactobacillus strains on a human colorectal adenocarcinoma cell line (HT-29). The strains assessed were Limosilactobacillus (L.) reuteri VA 102, Ligilactobacillus (L.) animalis VA 105, and Limosilactobacillus (L.) reuteri KCTC 3594 (ATCC 23272). The viability of HT-29 cells was evaluated using the MTT assay. The findings revealed that cell-free supernatants (CFS) exhibited significant anticancer effects. Heat-inactivated L. reuteri VA 105 and L. reuteri KCTC 3594 induced a pronounced reduction in cell viability. Furthermore, live cultures of L. reuteri VA 105 and L. reuteri VA 102 also showed reduced cell viability compared to the control group. These results suggest that CFS and heat-inactivated cells may be more suitable for therapeutic applications than live bacteria owing to their improved safety profiles and reduced potential for adverse effects. Our findings also emphasize the potential anticancer benefits of these LAB strains.

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 optimization of a prevention program utilizing a novel trivalent inactivated Salmonella bacterial vaccine to protect poultry from Salmonella infections was evaluated in this study. A total of 50 brown nick layers were divided into 5 groups, A to E, each containing 10 hens. Group B hens were immunized with the SG9R vaccine, group C chickens were immunized with the trivalent inactivated Salmonella bacterial vaccine, and group D hens were primed with SG9R and boosted with the trivalent inactivated Salmonella bacterial vaccine. Group E hens were injected with sterile PBS. All hens in groups B to E were orally challenged with a mixture of wild-type S. Enteritidis, S. Typhimurium, and S. Gallinarum (approximately 6×10⁹ CFU/ 0.2 ml/bird). Serum IgG titers, CD3+CD4+ T-cell levels, and CD3+CD8+ T-cell levels in group D were significantly higher than those in group A. Additionally, all animals in groups A to D showed no clinical symptoms and survived after the virulent challenges, whereas all chickens in group E died following the challenge. The challenge strains of S. Enteritidis, S. Typhimurium, and S. Gallinarum were not isolated from the liver, spleen, cecum, and cloaca of group D chickens. These findings indicate that priming with SG9R and boosting with a trivalent inactivated Salmonella bacterial vaccine can be an effective approach for preventing Salmonella infections by inducing robust protective humoral and cellular immune responses in chickens.

Transplantation is considered to be a very useful approach to improve human welfare and to prolong life-span. Heterologous organ transplantation using pig organs which are similar to human beings and easy to make mass-production has known as one of the alternatives. To ensure potential usage of the pig organ for transplantation application, it is essentially required to generate transgenic pig modifying immuno-related genes. Previously, we reported production of heterozygous α 1,3-galactosyltransferase (GalT) knock-out and human membrane cofactor protein (MCP) expressing pig (GalT-MCP/+), which is enforced for suppression of hyperacute and acute immunological rejection. In this study, we reported generation of homozygous pig (GalT-MCP/-MCP) by crossbreeding GalT-MCP/+ pigs. Two female founders gave birth to six of GalT-MCP/-MCP, and seven GalT-MCP/+ pigs. We performed quantitative real-time PCR, western blot, and flow cytometry analyses to confirm GalT and MCP expression. We showed that fibroblasts of the GalT-MCP/-MCP pig do not express GalT and its product Gal antigen, while efficiently express MCP. We also showed no expression of GalT, otherwise expression of MCP at heart, kidney, liver and pancreas of transgenic pig. Taken together, we suggest that the GalT-MCP/-MCP pig is a useful candidate to apply xenotransplantation study.