花生DNA快速简便提取方法的研究

在花生新鲜针叶和干叶ＤＮＡ提取过程中,应用ｐＨ值较低,无机盐浓度较高的提取缓冲液可沉淀蛋白质,其中加入２％的β－巯基乙醇可有效地防止次生代谢物质使ＤＮＡ变色。提取出的ＤＮＡ样品产率在９２．６～２１６．３１ｎｇ／ｍｇ．ｆｗ,ＤＮＡ质量和纯度较高,２６０ｎｍ／２８０ｎｍ光密度比值在１．８～１．９之间。所得ＤＮＡ可直接用于限制性内切酶酶切,并可用于随机引物ＰＣＲ扩增。该方法为花生分子生物学研究提供基础。     

Feed endoxylanase type and dose affect arabinoxylan hydrolysis and fermentation in ageing broilers

Despite the general use of endoxylanases in poultry feed to improve broiler performance, the abundance of different endoxylanase products and the variable response to their application in the field prevent a clear understanding of endoxylanase functionality in vivo. To gain insight into this functionality, we investigated the impact of endoxylanase type (Belfeed from Bacillus subtilis versus Econase XT from Nonomuraea flexuosa) and dose (10, 100, 1,000 mg/kg) in combination with broiler age on arabinoxylan (AX) hydrolysis and fermentation in broilers (Ross 308) fed a wheat-soy based diet. In a digestibility trial and a performance trial, a total of 1,057 one-day-old chicks received the control diet or 1 of the 6 endoxylanase supplemented wheat-soy based diets with, respectively, 5 replicate cages and 8 replicate pens per dietary treatment per trial. The AX content and structure, the AX digestibility values and the short-chain fatty acids produced were analysed at the level of the ileum, caeca and excreta at d 11 and 36. Endoxylanase supplementation resulted in a more extensive solubilisation of wheat AX and a reduction in the intestinal viscosity compared to the control (P < 0.05). A high endoxylanase dose was, however, required to obtain increased hydrolysis of the dietary AX along the gastrointestinal tract against the control (P < 0.001). Depending on the type of endoxylanase, a pool of AX with distinct physicochemical properties was created. The B. subtilis endoxylanase created a large pool of soluble AX in the ileum, thereby increasing ileal viscosity compared to broilers fed an endoxylanase from N. flexuosa (P < 0.001). The N. flexuosa endoxylanase mainly triggered caecal AX fermentation in young broilers, by delivering easily fermentable AX substrates with a low degree of polymerisation (P = 0.03). The effects were particularly present in young broilers (d 11). From this study, it is clear that the type and dose of endoxylanase added to wheat-soy based diets determine the nature of AX substrates formed. These, in turn, affect the intestinal viscosity and the interplay between the dietary AX compounds and microbiota, hence dictating AX digestion at young broiler ages and performance outcomes towards slaughter age.     

Comparison of New Ultrasonic Digestion Approaches for Plant Matrices in the Analysis of Trace Metals by Inductively Coupled Plasma–Optical Emission Spectroscopy Analysis: Contrast with USEPA Method 3050B

An ultrasonic method using two approaches, A and B, along with a reference Environmental Protection Agency (EPA) Method 3050B [i.e., a mixture of 30 mL of nitric acid–hydrochloric acid–hydrogen peroxide–water (HNO₃-HCl-H₂O₂-H₂O)] were contrasted for leaching of a plant matrix. The trace metals were arsenic, cadmium, cobalt, chromium, copper, mercury, manganese, nickel, lead, selenium, and zinc (As, Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb, Se and Zn) and quantified by ICP-OES followed by an investigation into residue formation and the impact of digestion time. Approach B was the most accurate and precise with percent recoveries ranging between 99 and 120%, whereas ultrasonic approach A and the USEPA method 3050B gave similar results with poor accuracies and precisions. In the optimization of the digestion time using approach B, the total metal recovery was fairly the same over a period of 120 min except for Cr and Cu, which showed slight variations.     

桑叶在水产饲料中应用研究进展

随着水产养殖业集约化和规模化的发展,全球鱼粉需求和供应之间的矛盾加大,急需寻找新的蛋白源,保障水产养殖业健康可持续发展。桑叶作为一种新型植物蛋白原,含有丰富的蛋白质、维生素、矿物质和天然活性物质,并且氨基酸种类齐全,营养价值高,具有较大的开发潜力。本文结合近几年桑叶的研究报道,介绍了桑叶的营养价值,综述了饲料中使用桑叶粉对水产动物的生长、消化、品质和原料表观消化率的影响,初步探讨了提高桑叶在水产饲料中应用的可行性。     

大豆蛋白酶解物的抗消化性及离子强度对其聚集的影响

分别采用体积排阻色谱和Folin-酚法测定了经酸性蛋白酶(蛋白酶M、胃蛋白酶)、中性蛋白酶(蛋白酶A、蛋白酶N)、枯草杆菌碱性蛋白酶Alcalase酶解后制得的大豆肽经过模拟体内消化之后分子量的变化,以及离子强度对酶解物聚合的影响.大豆肽的高效液相色谱体积排阻色谱图显示,50％左右的大豆肽的分子量集中在10～30 kD...     

配合饲料饲喂育成期水貂消化试验及饲养效果观测

将68只水貂按雌雄、体质量随机分为两组,分别饲喂常规鲜料(对照组)和配合饲料(试验组),在育成期进行饲养试验和消化试验。结果表明:母貂干物质和粗蛋白表观消化率对照组均高于试验组(P<0.05),公貂干物质和粗蛋白表观消化率对照组均显著高于试验组(P<0.01);试验组水貂干物质采食量和可消化干物质均略高于对照组;粗脂肪和粗纤维表观消化率试验组均高于对照组,差异极显著(P<0.01);对照组和试验组水貂能量表观消化率差异不显著(P>0.05);试验组水貂质量的增加均大于对照组。试验结果证实,采用配合饲料与一定比例的新鲜原料喂养水貂,是提高水貂生长性能、解决其饲料来源困难的比较合理的办法。     

Novel insecticide polymer chemistry to reduce the enzymatic digestion of a protein pesticide, trypsin modulating oostatic factor (TMOF)

A limiting factor in the use of proteins as insecticides, especially when the site of action is in the insect hemocoel, is protease degradation in the digestive system and hemolymph and movement across the midgut ventriculus. Trypsin modulating oostatic factor (TMOF) is a per os mosquito peptidic larvacide which moves across the digestive system and binds to receptors on the hemolymph side of the gut where the hormone inhibits protease synthesis and food utilization ultimately causing death. In the current study, the in vitro degradation of TMOF by the digestive enzyme, leucine aminopeptidase, was inhibited by conjugation of TMOF-K with aliphatic polyethylene glycol (PEG) polymers. Structure activity studies demonstrated a correlation between the molecular weight of the PEG polymer and resistance to digestion and show proof of concept that aliphatic-PEG protein polymerization can be used to prevent protease degradation of a protein insecticide.     

Young rice protein as a new source of low allergenic plant-base protein

New plant-based milk protein substitutes that do not exhibit immune-mediated adverse effects are in high demand. In-vitro digestibility (oral, gastric and duodenal) of rice was investigated at flowery, milky, dough and mature stages using SDS-PAGE and band densitometry. Possible allergenic proteins found were 13–14, 22–23, 37–39, and 52 kDa. Protein content in young rice at flowery-to-milky stage (9.19–11.48%) was higher than rice bran (8.90%). Possible allergens of young rice rapidly decreased and completely disappeared after digestion. Digestibility of dough-to-mature stage was much slower especially for colored rice. Proteins at 13–14, 52 kDa and 13–14, 37–39 kDa were indigestible in the mature stage of PT1 and RB respectively. High protein content and low allergic potential of young rice supported its possible use as a new plant-based protein alternative.     

限制性内切酶在PCR缓冲液中酶切活性的研究(英文)

选取6 种常用的限制性内切酶(Acc I、Ban II、EcoR I、Hind III、Sac I、Sca I)用于检测其直接在PCR 缓冲液中完全酶切DNA 的活性。结果表明:在PCR 缓冲液中,需要加入MgCl2作为激活因子的前提条件下,除了EcoR I 表现出星状活性外,其他酶均能完全酶切无甲基化λDNA,其结果与在酶生产厂商推荐的反应缓冲液中进行的酶切反应结果一致;在完成PCR 扩增的PCR 缓冲液中,所有酶(包括RcoR I)均能功能正常地对PCR 产物完全酶切,表明可在PCR 扩增完毕后的反应体系中直接加入限制性内切酶进行酶切反应,无需对PCR 产物进行预先纯化;镁离子浓度在2.5 mmol·L-1至10 mmol·L-1范围内对PCR 缓冲液中的限制性内切酶的活性无显著影响。上述方法可用于限制性片段长度多态(RFLP)以及单链构像多态(SSCP)研究中,但能否用于分子克隆有待进一步实验。图3 表1 参7。     

Marine wax ester digestion in salmonid fish: a review

Alternative marine resources from lower trophic levels could partly cover the rapidly increasing needs for marine proteins and oils in the future. The North Atlantic calanoid copepod, Calanus finmarchicus, has a high level of lipids rich in n‐3 fatty acids. However, these animals have wax esters as the main lipid storage component rather than triacylglycerol (TAG). Although these esters are considered difficult to digest by many fish, is it well known that juvenile Atlantic salmon (Salmo salar) feed on zooplankton species. It is therefore possible that the capacity to utilize these lipids should be well developed in salmonids. Nonetheless, salmon hydrolyse wax esters slower than TAG and absorb fatty alcohols slower than fatty acids. However, salmon have several adaptations to digest diets rich in wax esters. These includes increased feed conversion, higher production of bile and higher activity of lipolytic enzymes in the midgut. Atlantic salmon has been shown to feed and grow on diets with a medium amount of wax esters (30% of the lipid) with results comparable to fish maintained on fish oil diets. Ingestion of higher level of wax esters (50% of the lipid) cause, however, poorer lipid digestibility and growth, so that optimal utilization of wax esters in Atlantic salmon is closer to 30% than 50% of the dietary lipid.