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Aflatoxin B1 (AFB1) and zearalenone (ZEN) exert deleterious effects to human and animal health. In this study, the ability of a CotA laccase from Bacillus subtilis (BsCotA) to degrade these two mycotoxins was first investigated. Among the nine structurally defined chemical compounds, methyl syringate was the most efficient mediator assisting BsCotA to degrade AFB1 (98.0%) and ZEN (100.0%). BsCotA could also use plant extracts, including the Epimedium brevicornu, Cucumis sativus L., Lavandula angustifolia, and Schizonepeta tenuifolia extracts to degrade AFB1 and ZEN. Using hydra and BLYES as indicators, it was demonstrated that the degraded products of AFB1 and ZEN using the laccase/mediator systems were detoxified. Finally, a laccase of fungal origin was also able to degrade AFB1 and ZEN in the presence of the discovered mediators. The findings shed light on the possibility of using laccases and a mediator, particularly a natural plant-derived complex mediator, to simultaneously degrade AFB1 and ZEN contaminants in food and feed.

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Total five ear skin fibroblast lines (named F1, F2, F3, F4 and F5) from different newborn Holstein cows have been used as nuclear donor cells for producing cloned cows by somatic cell nuclear transfer (SCNT). The effects of these cell lines on both in vitro and in vivo developmental rates of cloned embryos, post-natal survivability and incidence of large offspring syndrome (LOS) were examined in this study. We found that the different cell lines possessed the same capacity to support pre-implantation development of cloned embryos, the cleavage and blastocyst formation rates ranged from 80.2 ± 0.9 to 84.5 ± 2.5% and 28.5 ± 0.9 to 33.3 ± 1.4%, respectively. However, their capacities to support the in vivo development of SCNT embryos showed significant differences (p < 0.05). The pregnancy rates at 90 and 240 day were significantly lower in groups F2 (4.9% and 3.3%) and F3 (5.4% and 5.4%) compared to groups F1 (23.3% and 16.3%), F4 (25.7% and 18.6%) and F5 (25.9% and 19.8%) (p < 0.05). The cloning efficiency was significantly higher in group F5 than those in group F1, F2, F3 and F4 (9.3% vs 4.1%, 1.2%, 2.0% and 5.0%, respectively, p < 0.05). Moreover, large offspring syndrome (LOS) incidence in group F5 was significantly lower than those in other groups (p < 0.05). All cloned offspring from cell line F1, F2, F3 and F4 showed LOS and gestation length delay, while all cloned offspring from F5 showed normal birthweight and gestation length. We concluded that the nuclear donor cell lines have significant impact on the in vivo development of cloned embryos and the incidence of LOS in cloned calves.

Forkhead box O 1 (FoxO1) is an important transcription factor implicated in adipogenesis. In this study, we detected the breed differences in FoxO1 between Bamei pigs (an obese breed) and Large White pigs (a lean breed). Compared with Large White pigs, the BW of Bamei pigs was lower (P < 0.01), but back fat thickness, fat percent, and intramuscular fat content were greater (P < 0.01). The levels of FoxO1 mRNA and protein were lower (P < 0.01) in subcutaneous adipose tissue (SAT) of Bamei pigs at 180 d, adipocytes and stromal-vascular fraction extracted from SAT of Bamei pigs at 1 d compared with Large White pigs. Knockdown of FoxO1 increased triglyceride content (P < 0.01) and upregulated the levels of adipocyte fatty-acid binding protein, PPARγ, and CCAAT enhancer-binding protein α (C/EBPα) at 6 d after porcine preadipocytes were induced. Furthermore, the transcriptional regulation of FoxO1 through C/EBPβ during early porcine preadipocyte differentiation and the effect of insulin on phosphoinositide 3 kinase (PI3K)/glycogen synthase kinase 3β (GSK3β) signal pathway by FoxO1 were examined. The results indicated that FoxO1 inhibited transcription activity of C/EBPβ, whereas C/EBPβ did not affect transcription activity of FoxO1. At 6 and 12 h of early differentiation, knockdown of FoxO1 triggered the transcription activity of C/EBPβ. In addition, FoxO1 protein interacted with C/EBPβ protein in porcine adipocytes at 12 h after induction. Under treatment with 100 nM insulin, knockdown or overexpression of FoxO1 mediated PI3K/GSK3β signaling via upregulating or downregulating the levels of GSK3β and its phosphorylation in adipocytes. Taken together, there is low, but detectable, expression of FoxO1 in SAT of obese pigs and FoxO1 inhibited adipogenesis through C/EBPβ and PI3K/GSK3β signaling pathway. These findings provide useful information to further the understanding of the function of FoxO1 in porcine adipogenesis.

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