15N-Urea Efficiency in Maize as Influenced by Humic Substances and Urease Inhibitors Treatments

Urea treated with urease inhibitors (UI) in association with humic substances (HS) is expected to improve urea efficiency by reducing ammonia losses and also provide the benefits of HS such as improve nitrogen (N) recovery by plants. Ammonia volatilization (AV), ¹⁵N recovery efficiency in the soil (NRE) and ¹⁵N use efficiency (NUE) were evaluated in maize (Zea mays L.) in a greenhouse pot trial. Treatments consisted of ¹⁵N-urea treated with UI (0, 0.4% boron (B) +0.15% copper (Cu), 0.64% B, and NBPT (N-(n-butyl) thiophosphoric triamide)) and three levels of HS (0%, 0.6%, and 1.2%). A control treatment (without N) was also included. N treatments were applied at V4 (vegetative leaf stage 4) on soil surface. HS was not efficient on AV reduction, NBPT had the greatest reduction in AV, and B had higher efficiency on AV reduction than Cu. At V8 (vegetative leaf stage 8), NBPT had the greatest NUE followed by B. At VT (tassel fully emerged), NUE from urea treated with NBPT reduced when increased levels of HS, and NBPT had the greatest NRE. UI in association with HS treated-urea does not reduce AV and it does not improve NUE by maize in vegetative stages.     

我国畜禽饲料资源中微量元素铜含量分布的调查

目的 研究我国不同地区间各种饲料原料中铜含量分布情况,以及我国畜禽基础饲粮中铜水平,从而为饲粮中合理添补铜提供依据。方法 对采自全国31个省、直辖市和自治区的7大类（谷物籽实、谷物籽实加工副产品、植物性蛋白饲料、动物性蛋白饲料、牧草类、秸秆类和矿物质饲料）37种饲料原料共3 903个饲料样品,经预处理后用MARS6高通量密闭微波消解系统进行微波消解,然后用IRIS Intrepid II等离子体发射光谱仪测定其铜含量。用国家标准物质猪肝粉或黄豆粉作为参照标准,以保证测定结果的可靠性。结果 饲料原料中铜含量测定结果表明：谷物籽实（包括玉米、小麦、稻谷和大麦）平均铜含量为3.95 mg·kg ^-1（范围为2.50—5.34 mg·kg ^-1）;谷物籽实加工副产品（包括玉米蛋白粉、玉米DDGS、玉米胚芽粕、次粉、小麦麸、小麦DDGS、碎米和米糠）平均铜含量为7.16 mg·kg ^-1（范围为1.62—12.13 mg·kg ^-1）;植物性蛋白饲料（包括膨化大豆、大豆粕、菜籽粕、棉籽粕、花生粕、亚麻粕和葵花粕）平均铜含量为16.37 mg·kg ^-1（范围为6.45—30.40 mg·kg ^-1）;动物性蛋白饲料（包括鱼粉、肉粉、水解羽毛粉、肠膜蛋白粉、血浆蛋白粉和血球蛋白粉）平均铜含量为11.14 mg·kg ^-1（范围为1.90—20.04 mg·kg ^-1）;牧草类（包括羊草、黑麦草、苜蓿和青贮玉米）平均铜含量为7.85 mg·kg ^-1（范围为4.31—9.92 mg·kg ^-1）;秸秆类（包括玉米秸秆、小麦秸秆、稻秸和甘薯藤）平均铜含量为7.50 mg·kg ^-1（范围为3.38—13.89 mg·kg ^-1）;矿物质饲料（包括石粉、磷酸氢钙、骨粉和贝壳粉）平均铜含量为6.79 mg·kg ^-1（范围为3.39—11.45 mg·kg ^-1）。这37种饲料原料的平均铜含量范围为1.62—30.40 mg·kg ^-1,而各类饲料原料铜含量分布规律是：植物性蛋白饲料（16.37 mg·kg ^-1）＞动物性蛋白饲料（11.14 mg·kg ^-1）＞牧草类饲料（7.85 mg·kg ^-1）＞秸秆类饲料（7.50 mg·kg ^-1）＞谷物籽实加工副产品（7.16 mg·kg ^-1）＞矿物质饲料（6.79 mg·kg ^-1）＞谷物籽实（3.95 mg·kg ^-1）。以省（区）为单位比较,发现不同地区间的玉米、小麦和大豆粕的铜含量差异显著（P＜0.05）。四川省玉米和大豆粕铜含量最高（分别为2.97和15.74 mg·kg ^-1）,内蒙古自治区最低（分别为1.66和11.72 mg·kg ^-1）;甘肃省小麦铜含量最高（5.61 mg·kg ^-1）,河北省最低（4.02 mg·kg ^-1）。根据全国各地猪、鸡常用的152个饲料配方计算出基础饲粮中可提供的铜含量为5.07—6.54 mg·kg ^-1,如根据我国及美国NRC猪、鸡饲养标准中铜营养需要量的要求,基础饲粮中的铜含量基本可提供猪的铜营养需要,可提供鸡大部分铜的营养需要,但上述估算尚未考虑不同饲料原料中铜的利用率。结论 我国不同种类和不同地区饲料原料中铜含量差异较大,全国猪、鸡常用基础饲粮配方中铜含量可提供猪、鸡大部分的铜营养需要量。因此,建议在配制饲粮时,应充分考虑不同地区基础饲粮中的铜含量及其利用率,精准配制饲粮,以满足畜禽高效生产需要,同时减少铜的添加和排放对环境的污染。     

内生贪铜菌的分离鉴定及其对铜污染含羞草生长的影响

【目的】研究贪铜菌及其与含羞草(Mimosa pudica)形成的共生体对重金属铜污染的抗性,为重金属污染环境的微生物修复提供参考,丰富功能菌株资源库。【方法】自含羞草根瘤中分离得到菌株HXC-8,扩增该菌株的16S rDNA序列,对其进行鉴定,并对其进行生理生化特性测定,研究菌株HXC-8对重金属铜的耐受性及去除率。此外,设置接菌组（含羞草种子在HXC-8菌悬液中浸泡6 h、自然阴干1 h）和未接菌组（含羞草种子在蒸馏水中浸泡6 h、自然阴干1 h）,将2组含羞草种子在Cu²⁺质量浓度分别为0,50,100,150,200,250,300,350,400 mg/L的混合营养液中培养,分析菌株HXC-8对铜胁迫下含羞草种子萌发和幼苗生长的影响。【结果】16S rDNA基因序列分析结果显示,菌株HXC-8为贪铜菌属（Cupriavidus sp.）细菌,将其命名为“贪铜菌HXC-8”。生理生化特性测定结果显示,贪铜菌HXC-8为革兰氏阴性菌,能分泌活性较高的H₂O₂酶和淀粉酶,不产生纤维素酶,不适合在肉汁培养基上生长,在糖代谢过程中无混合酸发酵,属于快生型内生菌。随着Cu²⁺质量浓度的升高,贪铜菌HXC-8的生长受到了抑制,该菌对Cu^2＋的耐受量是0～150 mg/L。当Cu^2＋质量浓度为25~150 mg/L时,随着菌液中Cu^2＋质量浓度的增大,贪铜菌HXC-8对铜的去除率逐渐降低,其中Cu^2＋质量浓度为25 mg/L时,贪铜菌HXC-8对铜的去除率最高,为97.93%。在0~400 mg/L Cu²⁺胁迫下,随Cu²⁺质量浓度的升高,未接菌组和接菌组含羞草种子萌发率均无显著变化,但种子活力指数明显下降;未接菌组和接菌组含羞草幼苗鲜质量、根长、茎长及株高均明显降低。当Cu²⁺质量浓度为0～300 mg/L时,接菌组与未接菌组含羞草幼苗鲜质量、根长、茎长、株高均无显著差异;当Cu²⁺质量浓度为350~400 mg/L时,接菌组幼苗以上指标均明显大于未接菌组。【结论】贪铜菌HXC-8对铜具有一定的耐受性,能够降低环境中游离态的Cu²⁺,可减缓重金属铜对含羞草生长的胁迫。     

水稻土溶解性有机质与Cu2+结合过程及机制研究

农田土壤中Cu²⁺的迁移转化对生物体健康产生重要影响,然而,水稻土中的溶解性有机质（DOM）与Cu²⁺的结合过程和机制尚不清楚。本研究通过动态光散射、三维荧光、同步荧光以及X射线光电子能谱（XPS）等方法,探究水稻土中DOM的粒径、光谱和结构特征及其与Cu²⁺的结合过程及机制。结果表明,土壤DOM的SUVA₂₅₄值为（3.25±0.07）L·（mg·m）^-1,低于4 L·（mg·m）^-1,其亲水性较高同时芳香性偏低,并且其芳香结构中含有较多的含氧官能团。水稻土DOM的构成以类富里酸和类腐植酸为主,Cu²⁺与类腐植酸络合稳定常数（lgK_M=5.07）高于类富里酸（lgK_M=4.81）。表明稻田土壤DOM中的类腐植酸组分与Cu²⁺优先结合,尤其以短波类腐植酸中的酚基、羟基和酚羟基等含氧基团对Cu²⁺的添加表现得更为敏感。另外,DOM表面疏松的介孔结构、负电荷和羟基、酚羟基以及氨基等官能团对Cu²⁺吸附起到关键作用。     

Dietary copper supplementation modulates performance and lipid metabolism in meat goat kids

Forty-eight male Lezhi black goat kids with similar body weight ((12.09±1.70) kg) and age ((60±5) d) were used to determine the effect of dietary copper (Cu), in the form of reagent grade Cu sulfate (CuSO₄∙5H₂O), on performance, serum lipid profile, and the relative mRNA abundance of genes involved in lipid metabolism.  Goat kids were stratified by body weight and randomly assigned to one of 4 treatment groups.  Each treatment consisted of 12 replicate pens with each pen containing one goat kid.  Treatment groups received the basal diet with no supplemental Cu (control), basal diet plus 10 mg of Cu kg^–1 of dry matter (DM), basal diet plus 20 mg of Cu kg^–1 of DM, or basal diet plus 30 mg of Cu kg^–1 of DM.  Goats were housed individually in pens and fed a high-concentrate pelleted diet for 60 d.  Average daily gain, average daily feed intake and feed:gain of goats were not affected by dietary Cu supplementation (P>0.10).  No differences were detected in serum total cholesterol, triglyceride, and high density lipoprotein cholesterol concentrations of goat kids fed with different Cu concentrations (P>0.05).  However, serum low density lipoprotein cholesterol concentrations decreased linearly (P=0.01) as the concentration of dietary Cu increased.  Intramuscular fat content of longissimus muscle increased (P=0.002) quadratically and liver Cu concentrations increased (P<0.001) linearly as dietary Cu concentration increased.  Compared with the control, dietary supplementation of 20 mg Cu kg^–1 DM decreased the relative mRNA abundance of fatty acid-binding protein 4 (P=0.01) and lipoprotein lipase (P=0.05), and tended to decrease the relative mRNA abundance of carnitine palmitoyltransferase I (P=0.06) in longissimus muscle of goats.  The relative mRNA abundance of peroxisome proliferator-activated receptor alpha (P<0.001), carnitine acetyltransferase (P=0.001), and carnitine palmitoyltransferase I (P=0.001) were also decreased in liver by Cu supplementation.  These results indicate that dietary supplementation of Cu modified lipid metabolism by increasing muscular fat and decreasing serum low density lipoprotein cholesterol, and the modification might be associated with the reduction of relative mRNA abundance of genes for oxidation of long-chain fatty acid in muscle and liver of Lezhi black goat kids.     

不同剂量的蛋氨酸铜对肉鸡肝脏病理组织结构和粪便铜含量的影响

选择1日龄健康AA肉仔鸡120只,随机分成4组。使用蛋氨酸铜作为铜源,饲喂玉米-豆粕型基础日粮,对照组饲料铜含量为11mg/kg,3个高铜试验组分别为:110mg/kg、220mg/kg和330mg/kg,试验至60日龄结束。于第12,36,60日龄在各组内随机采血,测定血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)活性;测量60日龄各组肝脏和粪便的铜含量并观察肝脏组织病理变化。结果显示:①血清ALT和AST活性均在正常范围内,没有异常升高;②110mg/kg和220mg/kg组的肝脏组织中铜含量比对照组有所升高,但是差异不显著(P>0.05),而330mg/kg组则极显著高于其他3个试验组(P<0.01);110mg/kg,220mg/kg和330mg/kg组的粪便中铜含量均极显著高于对照组(P<0.01);③330mg/kg组的肝脏组织大部分细胞中度变性,其他各组病理变化不明显。试验结果表明,日粮中蛋氨酸铜浓度达到330mg/kg时,肝脏内的铜蓄积量会迅速增多且对肝脏造成较大的损伤,粪便中的铜含量随着日粮中铜含量的增加而迅速增多,可能对环境造成不利影响。     

枯草芽孢杆菌铜对先天性缺铜大鼠生长发育、器官指数、铜沉积量、血清生化和抗氧化指标的影响

本试验旨在研究枯草芽孢杆菌铜对先天性缺铜大鼠生长发育、器官指数、铜沉积量、血清生化和抗氧化指标的影响。试验分为模型期和干预试验期。模型期选取怀孕10 d的SD大鼠72只,平均分为模型组和对照组,每组6个重复,每个重复6只。模型组大鼠孕期饲喂缺铜饲粮(铜含量为0.33 mg/kg),对照组大鼠孕期饲喂正常饲粮(在缺铜饲粮基础上添加6.0 mg/kg五水硫酸铜),持续到哺乳期结束,验证建模是否成功。干预试验期选择24日建模成功的缺铜幼龄大鼠108只,随机分成6个试验组,Ⅰ～Ⅲ组为硫酸铜组(缺铜饲粮基础上分别添加3.0、6.0和9.0 mg/kg五水硫酸铜),Ⅳ～Ⅵ组为枯草芽孢杆菌铜组(缺铜饲粮基础上分别添加3.0、6.0和9.0 mg/kg枯草芽孢杆菌铜),每个组3个重复,每个重复6只。试验期35 d。结果表明:1)在铜添加量相同条件下,Ⅳ组体重和血清谷丙转氨酶(ALT)、谷胱甘肽过氧化物酶(GSH-Px)活性显著或极显著高于Ⅰ组(P<0.05或P<0.01),Ⅳ组血清谷草转氨酶/谷丙转氨酶(AST/ALT)显著低于Ⅰ组(P<0.05)。Ⅴ组血清铜蓝蛋白(CP)、碱性磷酸酶(AKP)活性显著或极显高于Ⅱ组(P<0.05或P<0.01),Ⅴ组血清丙二醛(MDA)含量显著低于Ⅱ组(P<0.05)。Ⅵ组血清AKP活性和肾脏中铜沉积量显著或极显高于Ⅲ组(P<0.05或P<0.01)。2)在硫酸铜组中,Ⅱ、Ⅲ组体重、体长显著高于Ⅰ组(P<0.05),Ⅲ组血清CP活性显著或极显著高于Ⅰ、Ⅱ组(P<0.05或P<0.01),Ⅲ组血清AKP活性显著高于Ⅱ组(P <0.05),Ⅲ组血清谷草转氨酶(AST)、GSH-Px和铜锌超氧化物歧化酶(CuZn-SOD)活性及肾脏中铜沉积量显著或极显著高于Ⅰ组(P<0.05或P<0.01),Ⅲ组血清M DA含量极显著低于Ⅰ组(P <0.01)。在枯草芽孢杆菌铜组中,Ⅵ组体重、体长,血清AST、GSH-Px和CuZn-SOD活性,心脏、肝脏、肾脏中铜沉积量显著或极显著高于Ⅳ组(P<0.05或P<0.01),Ⅵ组血清CP和AKP活性显著或极显著高于Ⅳ、Ⅴ组(P <0.05或P <0.01)。由此可见,枯草芽孢杆菌铜能提高先天性缺铜大鼠体重,血清CP、AKP、ALT活性,提高机体抗氧化能力,增加肝脏、肾脏中铜沉积量,效果优于硫酸铜,且降低了饲粮中铜添加量。     

The effect of copper nanoparticles and copper (II) salt on redox reactions and epigenetic changes in a rat model

The aim of the study was to evaluate the effects of a diet containing different levels of Cu in two different chemical forms (carbonate and nanoparticles) on redox reactions and epigenetic changes in a rat model. For 4 weeks, five experimental groups (eight rats in each) were fed diets with two dosages of added Cu (standard—6.5 mg/kg or half of the standard dosage—3.25 mg/kg, and as a negative control no additional Cu in the mineral mixture) in two forms (standard—CuCO₃ and copper nanoparticles). Addition of Cu nanoparticles resulted in higher Cp (ceruloplasmin) activity and LOOH (lipid peroxides) and MDA (malondialdehyde) content, as well as decrease the CAT (catalase) activity and level of PC (protein carbonyl), 3‐NT (3‐nitrotyrosine), 8‐OHdG (8‐hydroxydeoxyguanosine), GSH + GSSG (total glutathione) and DNA methylation. Reducing the dose of copper resulted in a decrease in the level of LOOH and GSH + GSSG as well as CAT activity, but increased the level of PC and methylated DNA. Based on these evidence, we concluded that addition of copper nanoparticles in the diet reduces protein oxidation and nitration as well as DNA oxidation and methylation. Lowering the level of Cu in the diet increases the oxidation of proteins and DNA methylation.     

铜对意大利蜜蜂工蜂生理机能的影响

本研究旨在探索意大利蜜蜂工蜂成蜂饲粮中铜的添加水平对蜜蜂生长发育、生理生化及抗氧化性能的影响。从蜂群中随机取3000只刚出房的蜜蜂,随机平均分成6组,分别命名为A、B、C、D、E、F,每组5个重复,每个重复100只蜜蜂;6组工蜂幼虫分别饲喂铜的添加水平为0.00、2.00、4.00、6.00、8.00、10.00μg/g的糖水,饲养至全部死亡。分别取成蜂测定体成分、血淋巴生化指标、抗氧化指标、相关基因表达量、制作中肠切片,并测定寿命。结果表明:(1)铜对蜜蜂寿命的影响显著,从蜜蜂存活率曲线图来看,当饲粮中铜的添加水平为2.00μg/g时对蜜蜂中肠食膜的结构最为有利。(2)与对照组相比,铜的添加水平为2.00~10.00μg/g组9日龄蜜蜂体组织铜含量显著升高(P<0.05),蜜蜂血淋巴中总蛋白(TP)和总胆固醇(TCHO)含量显著降低(P<0.05)。(3)9日龄蜜蜂蜂体超氧化物歧化酶(Cu/Zn-SOD)活性随着铜添加水平的提高出现先升高后降低的趋势,在2.00μg/g时活性最高(P<0.05)。(4)当饲粮中铜的添加水平在0.00~10.00μg/g的范围内不断升高时,CAT、Ctr1基因相对表达量显著增加(P<0.05)。由此得出饲粮中铜的添加水平对蜜蜂的生理机能影响显著。