低磷低鱼粉饲料中添加植酸酶和蛋白酶对草鱼生长性能和消化生理的影响

本试验旨在研究低磷低鱼粉饲料中添加植酸酶和蛋白酶对草鱼生长性能、消化酶活性、营养物质表观消化率和免疫力的影响。试验选取平均体重为9.66 g的健康草鱼630尾,随机分为7个组,每组3个重复,每个重复30尾鱼。7组草鱼饲喂如下试验饲料:正常磷(磷酸二氢钙添加量2.0%)正常鱼粉(鱼粉添加量3.0%)饲料(对照)、低磷(磷酸二氢钙添加量1.3%)正常鱼粉饲料、低磷正常鱼粉并添加0.03%植酸酶饲料、正常磷低鱼粉(鱼粉添加量1.5%)饲料、正常磷低鱼粉并添加0.05%蛋白酶饲料、低磷和低鱼粉饲料、低磷低鱼粉并添加0.03%植酸酶和0.05%蛋白酶饲料。饲养试验持续56 d。结果显示:与饲喂正常磷正常鱼粉饲料草鱼相比,饲喂低磷低鱼粉饲料草鱼的增重率、特定生长率、营养物质(干物质、蛋白质、钙和磷)表观消化率、肠道消化酶(胰蛋白酶、糜蛋白酶和淀粉酶)活性、血清溶菌酶活性均显著降低(P<0.05),饲料系数和血清丙二醛含量显著升高(P<0.05)。在低磷正常鱼粉饲料、正常磷低鱼粉饲料、低磷低鱼粉饲料中分别添加植酸酶、蛋白酶、植酸酶与蛋白酶后,草鱼的上述指标均得到显著改善(P<0.05)。以上结果表明,在低磷低鱼粉饲料中添加植酸酶和蛋白酶能够提高草鱼的营养物质表观消化率、肠道消化酶活性和免疫力,从而改善其生长性能。     

Expression of recombinant phytase of Bacillus subtilis E20 in Escherichia coli HMS 174 and improving the growth performance of white shrimp,Litopenaeus vannamei,juveniles by using phytase‐pretreated soybean meal‐containing diet

To produce a thermostable and neutral phytase (phy) of Bacillus subtilis E20 in Escherichia coli HMS174 and evaluate its efficiency in improving growth performance. The phy C of B. subtilis E20 was expressed in E. coli HMS 174, and then the 42‐kDa recombinant phy C was purified by Ni‐NAT and analysed by SDS–PAGE. The recombinant phy C had optimal ranges of pH of 6 ~ 7 and temperature of 50 ~ 60 °C. A thermostability analysis showed that the enzyme is a thermostable phytase, and around 33% of residual activity was detected after being incubated at 90 ~ 100 °C for 10 min. The recombinant phy C‐pretreated soybean meal for feed preparation improved white shrimp, Litopenaeus vannamei, growth and feed efficiency. Overall, the neutral and thermostable phy C is suitable for aquafeed, and it is able to improve the nutritional utilization, resulting in enhanced shrimp growth and reduced feed costs.     

Impact of reduced dietary crude protein levels and phytase enzyme supplementation on growth response,slurry characteristics,and gas emissions of growing pigs

This experiment was carried out to evaluate the effect of reduced dietary crude protein (CP) levels supplemented with or without exogenous phytase on growing pigs. Six dietary treatments arranged in a 3 × 2 factorial arrangements of 3 CP levels (containing 14%, 16%, and 18% CP) supplemented each with or without 5,000 FTU/g phytase enzyme. Thirty growing pigs (average weight of 17.80 ± 0.10 kg) were allotted to the six dietary treatments in a complete randomized design. The final weight, daily weight gain, and feed conversion ratio (FCR) increased significantly with increasing CP levels. While, phytase supplementation improved (p = .044) FCR in pigs. Total solid and volatile solid content of the slurry were higher (p = .001) in pigs fed 14% and 16% CP diets supplemented with phytase when compared with other treatment groups. Concentration of methane gas emitted was lowest (p = .001) in the slurry of pigs fed 14% CP diet with or without phytase and those fed 16% CP diet with phytase supplementation. In conclusion, reduction in dietary CP levels resulted in reduced weight gain and poor FCR. While, reduced CP with phytase supplementation reduced concentration of methane gas emitted.     

芽孢杆菌中性植酸酶基因的原核表达及酶学性质分析

植酸酶作为饲料添加剂能够有效提高动物对饲料中磷的利用率及减少粪便中磷排放对环境的污染,并降低植酸的抗营养作用。为了获得性能稳定的高活性植酸酶,采用PCR扩增芽孢杆菌(Bacillus sp.)中性植酸酶基因phyC(GenBank登录号:FJ986327)的成熟肽编码序列,将其克隆进原核表达载体pET-28a(+),并转化E.coli BL21(DE3)进行表达。在37℃条件下以0.5 mmol/L IPTG诱导4 h能够获得大量包涵体蛋白,在25℃条件下以0.5 mmol/L IPTG诱导6 h有利于可溶性蛋白的获得。利用Ni-NTA亲和层析柱纯化重组植酸酶产物,获得的中性植酸酶的部分酶学特性为:耐热性较好,最适反应温度55℃,在70℃处理10 min可保持20%以上的酶活性;耐酸、碱能力较强,最适pH 6.0～7.0,pH 5.5～9.0时能保持80%以上的酶活性,pH 5.0～10.0时处理60 min仍能保持70%以上的酶活性,在pH 2.0～4.0时能保持40%以上的酶活性。利用构建的切除芽孢杆菌中性植酸酶基因phyC信号肽编码序列的原核表达载体及优化的诱导表达条件,能够在大肠杆菌中高量表达性能稳定的芽孢杆菌中性植酸酶。     

玉米-杂粕型日粮中添加不同植酸酶对蛋鸡生产性能和蛋品质及钙磷代谢的影响

选取720只24周龄海兰褐蛋鸡,随机分为8个处理,每个处理6个重复,每个重复15只鸡。8个处理分别为正对照组(PC)饲喂有效磷为0.36%的正常基础日粮,负对照组(NC)饲喂有效磷为0.16%的低磷日粮,其余试验组是在NC组日粮基础上分别添加两种植酸酶A或B各125、250、500 U/kg。其中植酸酶A是从大肠杆菌中提取,植酸酶B是从里氏木霉中提取。试验期为12周。结果表明:与PC组比较,NC和B500组产蛋率、平均蛋重和日采食量显著降低(P0.05),其他各添加植酸酶组恢复到PC组水平,各添加植酸酶组料蛋比和蛋品质无显著变化(P0.05);与PC组相比,NC组胫骨灰分、胫骨磷和血浆磷含量显著降低(P0.05),添加2种植酸酶组(除B500组外)恢复到PC组水平,B500组血浆磷含量显著低于PC组(P0.05),各试验组胫骨钙含量和血浆钙含量与PC组无显著差异(P0.05)。结果提示,低磷玉米-杂粕型日粮中添加2种植酸酶都能够缓解低磷引起的负效应,但2种植酸酶的最佳添加量不同,本试验条件下日粮中添加125 U/kg的植酸酶B或250 U/kg的植酸酶A都能满足蛋鸡对无机磷的需求。     

Prokaryotic Expression and Polyclonal Antibody Preparation of σC Protein of Novel Duck Reovirus DH13 Strain

The aims of the experiment was to optimize the prokaryotic expression system of σC protein,prepare polyclonal antibody against σC protein of novel duck reovirus (NDRV),and evaluate the titer of the antibody.The σC gene of NDRV-DH13 strain was amplified by RT-PCR,ligated into pET-30a(+) and pET-32a(+) expression vector,constructed prokaryotic expression plasmid,which were transformed into E.coli BL21(DE3) and the expression of the σC protein were induced by IPTG.The proteins expression were analyzed by SDS-PAGE.The recombinant protein without His tag was purified by digestion,and the recombinant protein with His-tagged was purified by Ni-NTA column.Then the polyclonal antibody was obtained from rabbits which had been immunized by the purified protein without His tag.Anti-His-labeled mAb and NDRV-σC positive serum were used as primary antibodies to evaluate antibodies specificity,the antibodies titer was detected by indirect ELISA (iELISA).SDS-PAGE results showed that the molecular weight of the expression on recombinant proteins were 34 and 37 ku respectively,the proteins were highly expression.Western blotting showed that they had the specific reaction and the prepared antibodies had higher affinity with σC protein,the titer were about 1:25 600 by iELISA detection.This study successfully constructed and optimized the prokaryotic expression system of the polyclonal antibody against σC protein,laid a foundation for the further study of σC protein and the research of genetically engineered vaccine.     

添加植酸酶对生长猪玉米豆粕型饲粮养分消化利用率的影响

选用体重2 0 kg左右的杜×长×大三元杂去势公猪,随机分为4组,进行消化代谢试验,测定玉米豆粕型基础饲粮(Ⅰ组)和以植酸酶250 U/kg(Ⅱ组)、500 U/kg(Ⅲ组)、750 U/kg(Ⅳ组)取代基础饲粮中40%、60%、80%的磷酸氢钙的试验饲粮中的粗蛋白质等养分的消化利用率。采用全收粪法进行两期消化代谢试验。结果表明:与对照组(Ⅰ组)相比,Ⅱ、Ⅲ、Ⅳ组的蛋白质表观消化率分别提高了3.50%、2.45%、2.32%,组间差异不显著(P>0.05);蛋白质生物学价值分别提高了5.75%、6.29%、6.62%,组间差异不显著(P>0.05);粗纤维表观消化率分别提高30.09%、54.10%(P<0.05)、13.07%;钙的表观消化率分别提高了4.02%、13.82%(P<0.05)、2.20%;磷的表观消化率分别提高了8.20%、47.36%(P<0.05)、13.39%;饲粮可消化能分别提高了1.89%、7.69%(P<0.05)、4.83%。     

Effects of step-up and step-down phytase regimens on performance and processing yields of male broilers from 1 to 35 d of age

An experiment was conducted to evaluate increasing or decreasing concentrations of dietary phytase on growth performance and processing yields of male broilers from 1 to 35 d of age. Treatments consisted of a positive control, a negative control (NC; less 0.14% Ca, 0.13% nonphytate P, and 0.03% Na), and 6 additional treatments based on the NC supplemented with phytase. Treatments 3 through 5 consisted of the NC diet supplemented with 500 phytase units (FTU)/kg of phytase in the starter phase that was either continued throughout the remainder of the study (treatment 3) or increased to 1,500 FTU/kg beginning in the finisher (treatment 4) or grower (treatment 5) phases. Treatment 6 had 1,500 FTU/kg of phytase throughout the study. Treatments 7 and 8 had 1,500 FTU/kg in the starter and decreased to 500 FTU/kg in the finisher or grower phases, respectively. At 35 d of age, broilers fed diets containing 1,500 FTU/kg of phytase had increased BW gain compared with birds fed diets formulated to contain 500 FTU/kg of phytase. Increasing phytase concentration between the starter and grower phases or decreasing phytase concentration between the grower and finisher phases negatively affected FCR from 1 to 35 d of age. Phytase supplementation did not affect weight and yield of carcass characteristics. Therefore, dietary phytase concentration should not be varied throughout production for optimum growth performance.     

Phylogenetic lineages in the Capnodiales

The Capnodiales incorporates plant and human pathogens, endophytes, saprobes and epiphytes, with a wide range of nutritional modes. Several species are lichenised, or occur as parasites on fungi, or animals. The aim of the present study was to use DNA sequence data of the nuclear ribosomal small and large subunit RNA genes to test the monophyly of the Capnodiales, and resolve families within the order. We designed primers to allow the amplification and sequencing of almost the complete nuclear ribosomal small and large subunit RNA genes. Other than the Capnodiaceae (sooty moulds), and the Davidiellaceae, which contains saprobes and plant pathogens, the order presently incorporates families of major plant pathological importance such as the Mycosphaerellaceae, Teratosphaeriaceae and Schizothyriaceae. The Piedraiaceae was not supported, but resolves in the Teratosphaeriaceae. The Dissoconiaceae is introduced as a new family to accommodate Dissoconium and Ramichloridium. Lichenisation, as well as the ability to be saprobic or plant pathogenic evolved more than once in several families, though the taxa in the upper clades of the tree lead us to conclude that the strictly plant pathogenic, nectrotrophic families evolved from saprobic ancestors (Capnodiaceae), which is the more primitive state.Taxonomic novelties: Brunneosphaerella Crous, gen. nov., B. jonkershoekensis (Marinc., M.J. Wingf. & Crous) Crous, comb. nov., B. protearum (Syd. & P. Syd.) Crous, comb. nov., Devriesia hilliana Crous & U. Braun, sp. nov., D. lagerstroemiae Crous & M.J. Wingf., sp. nov., D. strelitziicola Arzanlou & Crous, sp. nov., Dissoconiaceae Crous & de Hoog, fam. nov., Hortaea thailandica Crous & K.D. Hyde, sp. nov., Passalora ageratinae Crous & A.R. Wood, sp. nov., P. armatae Crous & A.R. Wood, sp. nov., Rachicladosporium cboliae Crous, sp. nov.