L-Leucine In Ovo Administration Causes Growth Retardation and Modifies Specific Amino Acid Metabolism in Broiler Embryos

L-Leucine (L-Leu) in ovo administration was demonstrated to afford thermotolerance and modified amino acids metabolism in post-hatched broiler chicks under heat stress. This study aimed to investigate the changes in embryonic growth and amino acid metabolism after in ovo injection of L-Leu. Fertilized broiler eggs were subjected to in ovo injection of sterile water or L-Leu on embryonic day (ED) 7. The weight of embryos and yolk sacs were measured on ED 12, 14, 16, and 18. Plasma and livers were collected on ED 14 and 18 for free amino acid analysis. The weight and relative weight of embryos were significantly lowered by in ovo administration of L-Leu, but those of yolk sacs were not altered. Moreover, L-Leu in ovo injection significantly reduced the plasma proline concentration during embryogenesis and increased the plasma concentrations of tyrosine (Tyr) and lysine (Lys) in ED 18. Hepatic Lys concentration was also significantly increased by L-Leu in ovo injection. Interestingly, Leu concentrations in the plasma and liver were not affected by L-Leu administration. These results indicated that in ovo administered L-Leu was metabolized before ED 14 and affected embryonic growth and amino acid metabolism during embryogenesis.     

L-亮氨酸产生菌的代谢工程改造及其发酵效率

以选育的获得产L-亮氨酸的谷氨酸棒杆菌MD106为出发菌株,通过重叠延伸PCR及自杀载体介导的同源重组技术构建panBC及alaT双基因缺失的突变株MD106ΔpanBC/ΔalaT,并对出发菌及双缺失重组菌进行摇瓶发酵试验,测定发酵指数。结果显示,发酵40h后,突变株的L-亮氨酸的产量为7.91g/L,比出发菌株提高43.3%,而且主要杂酸——丙氨酸减少超过80%,总杂酸比例较出发菌株减少55.6%。     

Effects of ＂Bioactive＂ amino acids leucine, glutamate, arginine and tryptophan on feed intake and mlRNA expression of relative neuropeptides in broiler chicks

Feed intake control is vital to ensuring optimal nutrition and achieving full potential for growth and development in poultry. The aim of the present study was to investigate the effects of L-leucine, L-glutamate, L-tryptophan and L-arginine on feed intake and the mRNA expression levels of hypothalamic Neuropeptide involved in feed intake regulation in broiler chicks. Leucine, glutamate, tryptophan or arginine was intra-cerebroventricularly (ICV) administrated to 4d-old broiler chicks respectively and the feed intake were recorded at various time points. Quantitative PCR was performed to determine the hypothalamic mRNA expression levels of Neuropeptide Y (NPY), agouti related protein (AgRP), pro-opiomelanocortin (POMC), melanocortin receptor 4 (MC4R) and corticotrophin releasing factor (CRF). Our results showed that ICV administration of L-leucine (0.15 or 1.5 μmol) significantly (P<0.05) increased feed intake up to 2 h post-administration period and elevated both hypothalamic NPY and AgRP mRNA expression levels. In contrast, ICV administration of L-glutamate (1.6μmol) significantly (P<0.05) decreased feed intake 0.25, 0.5 and 2 h post-injection, and increased hypothalamic CRF and MC4R mRNA expression levels. Meanwhile, both L-tryptophan (10 or 100μg) and L-arginine (20 or 200μg) had no significant effect on feed intake. These findings suggested that L-leucine and L-glutamate could act within the hypothalamus to influence food intake, and that both orexigenic and anorexigenic Neuropeptide genes might contribute directly to these effects.     

水稻对外源氨基酸的吸收与转运

采用不同浓度的L-蛋氨酸培养液培养秧苗,并用L-亮氨酸15N喷施分蘖期、抽穗期和蜡熟期水稻叶面,研究水稻根系对L-蛋氨酸、水稻叶面对L-亮氨酸15N吸收情况。结果表明,用1 000、2 000、3 000、4 000 mg/L的L-蛋氨酸溶液浸根培养,与对照相比,秧苗L-蛋氨酸含量分别增加8.2%,9.8%、19.7%、21.3%;用500 mg/L的L-亮氨酸15N在分蘖期、抽穗期和蜡熟期喷施水稻叶面,三个时期都会不同程度吸收L-亮氨酸,吸收最好的时期是抽穗期和蜡熟期。     

Effects of "Bioactive" amino acids leucine, glutamate, arginine and tryptophan on feed intake and mRNA expression of relative neuropeptides in broiler chicks

ABSTRACT: Feed intake control is vital to ensuring optimal nutrition and achieving full potential for growth and development in poultry. The aim of the present study was to investigate the effects of L-leucine, L-glutamate, L-tryptophan and L-arginine on feed intake and the mRNA expression levels of hypothalamic Neuropeptide involved in feed intake regulation in broiler chicks. Leucine, glutamate, tryptophan or arginine was intra-cerebroventricularly (ICV) administrated to 4d-old broiler chicks respectively and the feed intake were recorded at various time points. Quantitative PCR was performed to determine the hypothalamic mRNA expression levels of Neuropeptide Y (NPY), agouti related protein (AgRP), pro-opiomelanocortin (POMC), melanocortin receptor 4 (MC4R) and corticotrophin releasing factor (CRF). Our results showed that ICV administration of L-leucine (0.15 or 1.5 mumol) significantly (P < 0.05) increased feed intake up to 2 h post-administration period and elevated both hypothalamic NPY and AgRP mRNA expression levels. In contrast, ICV administration of L-glutamate (1.6 mumol) significantly (P < 0.05) decreased feed intake 0.25, 0.5 and 2 h post-injection, and increased hypothalamic CRF and MC4R mRNA expression levels. Meanwhile, both L-tryptophan (10 or 100 mug) and L-arginine (20 or 200 mug) had no significant effect on feed intake. These findings suggested that L-leucine and L-glutamate could act within the hypothalamus to influence food intake, and that both orexigenic and anorexigenic Neuropeptide genes might contribute directly to these effects.     

L-亮氨酸发酵培养基及基本发酵条件的优化

以黄色短杆菌FY-18为出发菌株,利用摇瓶发酵生产L-亮氨酸,并利用正交试验和单因素试验分别对其发酵培养基和基本发酵条件进行研究,从而优选出最佳培养基组分和培养条件参数。结果表明,L-亮氨酸发酵培养基的最适配比为:葡萄糖160 g/L、豆粕水解液20 g/L、玉米浆20 g/L、毛发粉15 g/L、硫酸铵70 g/L;其发酵最佳培养条件为:初始p H 7.2,培养温度32℃,发酵接种量为10%。在上述最佳条件下摇瓶中发酵的产酸量为22.5 g/L。