The present study was conducted to investigate the efficacy of oral administration of L. reuteri on growth performance, intestine histomorphology, immunological and gut microbiome of broilers. A total of twenty healthy chickens were used in a five-week experimental trial. Birds were assigned into one of two groups with orally administrated L. reuteri probiotic and without probiotic-(Control -Phosphate-buffered saline). A significant (p<0.05) body weight gain was observed in the chickens in L. reuteri treatment group compare to those in the control group at the end of the trial. In addition, the serum IGF-1 cytokines level significantly enhanced in L. reuteri treatment group. However, there were no notable effects observed on the villus height, crypt depth, muscularis thickness, and submucosal thickness in chickens orally inject with and without L. reuteri. At the phylum level, the presence of Firmicutes (99.5%) was highly abundantin the L. reuteri treatment group. Moreover, the fecal microbial communities of Lactobacillus (99.9%) showed average relative abundance at genus level in L. reuteri treatment group. From this, we concluded that oral administration of L. reuteri would be beneficial to enhance the body weight gain, gut microbiome, and immune status of broiler.

Summary In this study, we built on our previous research that discovered that autophagy activated the metaphase I stage during porcine oocytes in vitro maturation. We investigated the relationship between autophagy and oocyte maturation. First, we confirmed whether autophagy was activated differently by different media (TCM199 and NCSU-23) during maturation. Then, we investigated whether oocyte maturation affected autophagic activation. In addition, we examined whether the inhibition of autophagy affected the nuclear maturation rate of porcine oocytes. As for the main experiment, we measured LC3-II levels using western blotting after inhibition of nuclear maturation via cAMP treatment in an in vitro culture to clarify whether nuclear maturation affected autophagy. After autophagy inhibition, we also counted matured oocytes by treating them with wortmannin or a E64d and pepstatin A mixture. Both groups, which had different treatment times of cAMP, showed the same levels of LC3-II, while the maturation rates were about four times higher after cAMP 22 h treatment than that of the 42 h treatment group. This indicated that neither cAMP nor nuclear status affected autophagy. Autophagy inhibition during in vitro oocyte maturation with wortmannin treatment reduced oocyte maturation rates by about half, while autophagy inhibition by the E64d and pepstatin A mixture treatment did not significantly affect the oocyte maturation. Therefore, wortmannin itself, or the autophagy induction step, but not the degradation step, is involved in the oocyte maturation of porcine oocytes. Overall, we propose that oocyte maturation does not stand upstream of autophagy activation, but autophagy may exist upstream of oocyte maturation.

Busulfan is an alkylating agent that is commonly used as a preconditioning drug in the clinical transplantation of hematopoietic stem cells, and is also traditionally used method for chemical sterilization (McCune and Holmberg, 2009;Hur et al., 2017). The side effects of busulfan include bone marrow suppression, pulmonary fibrosis, and skin pigmentation. Previous studies have demonstrated that preconditioning using busulfan is more effective than irradiation during human B cell differentiation (Krivoy et al., 2008;Hayakawa et al., 2009). Additionally, busulfan is most commonly used for chemical sterilization during the transplantation of male germ cells. Busulfan depletes male germ cells and disrupts the junctions between Sertoli cells (Bucci and Meistrich, 1987).Numerous studies have reported that the dose of busulfan necessary for sterilization varies widely among species. The dose of busulfan necessary for sterilization in pigs, monkeys, coyotes, mice, rats and chickens is 40-

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