This study was conducted to investigate the effect of in ovo feeding (IOF) of folic acid on the folate metabolism, immune function and the involved epigenetic modification of broilers. A total of 400 (Cobb) hatching eggs were randomly divided into four groups (0, 50, 100 and 150 µg injection of folic acid at embryonic age 11 d), and chicks hatched from each treatment were randomly divided into six replicates with 12 broilers/replicate after incubation. The results indicated that, in ovo, 100- and 150-µg folic acid injections improved the hatchability. The average daily gain and feed conversion ratio increased in the 150-µg group during the late growth stage. Simultaneously, in the 100- and 150-µg groups, an increase was observed in hepatic folate content and the expression of methylenetetrahydrofolate reductase (d1 and 42) and methionine synthase reductase (d21). IgG and IgM concentrations, as well as plasma lysozyme activity of broilers, showed a marked increase along with increasing folic acid levels. The splenic expression levels of IL-2 and IL-4 were up-regulated, whereas that of IL-6 was down-regulated, in the 100- and 150-µg folic acid treatment groups. In addition, histone methylation in IL-2 and IL-4 promoters exhibited an enrichment of H3K4m2 but a loss of H3K9me2 with the increased amount of folic acid additive. In contrast, a decrease in H3K4m2 and an increase in H3K9me2 were observed in the IL-6 promoter in folic acid treatments. Furthermore, in ovo, the 150-µg folic acid injection improved the chromatin tightness of the IL-2 and IL-4 promoter regions. Our findings suggest that IOF of 150 µg of folic acid can improve the growth performance and folate metabolism of broilers, and enhance the relationship between immune function and epigenetic regulation of immune genes, which are involved with the alterations in chromatin conformation and histone methylation in their promoters.

The effect of dietary zinc deficiency on γ-glutamyl hydrolase (EC 3.4.22.12) activity and on pteroylpolyglutamate absorption was investigated in rats. Enzyme activity was determined in pancreas and gut lumen washings. Pteroylpolyglutamate absorption was studied by determining the rise in plasma folate levels following pteroylpolyglutamate ingestion. Two experiments were performed; in each purified diets were given to three groups of immature male Wistar rats for approximately 2 weeks. One group was given a Zn-deficient diet ad lib. (ZD), the second was pair-fed daily with this group on a Zn-adequate diet (PF) and the third was given the Zn-adequate diet ad lib. (AL). In Expt 1, significantly reduced pancreatic γ-glutamyl hydrolase activity was observed in ZD rats. In Expt 2, pteroylpolyglutamate was administered on day 14 and in the 3 h period following pteroylpolyglutamate ingestion, lumen γ-glutamyl hydrolase activity and plasma folate levels were significantly lower in ZD rats. Pancreas is reported as the source of lumen γ-glutamyl hydrolase in rats. The results presented indicate that the pancreatic enzyme is Zn-sensitive. It was concluded that, as a result, γ-glutamyl hydrolase activity was reduced in the lumen of ZD rats. Consequently the hydrolysis and subsequent absorption of pteroylpolyglutamate was impaired in ZD rats, as indicated by the smaller rise in plasma folate levels that occurred following pteroylpolyglutamate ingestion. Results of this study concur with previous observations in human beings and rats that Zn deficiency has an adverse effect on folate metabolism.