Effects of dietary Bacillus coagulans and yeast hydrolysate supplementation on growth performance,immune response and intestinal barrier function in weaned piglets

The present study investigated the effects of Bacillus coagulans and yeast hydrolysate supplementation on growth performance, immune response and intestinal barrier function of weaned piglets. Twenty-four weaned piglets with an average body weight (BW) of 6.89 ± 0.15 kg were divided into four diets for 28 days. The treatments were basal diet (control), basal diet supplemented with antibiotic (20 mg/kg colistin sulphate and 40 mg/kg bacitracin zinc, AT), probiotics (400 mg/kg Bacillus coagulans ≥5 × 109 CFU/g, BC) or yeast hydrolysate (5000 mg/kg yeast hydrolysate, YH). Average daily gain (ADG) and average daily feed intake (ADFI) were improved by AT and YH diets (p < 0.05), while BC diet only increased ADG (p < 0.05). The complement 3 (C3), lysozyme (LZM) and tumour necrosis factor-α (TNF-α) concentrations in serum were increased in BC diet (p < 0.05). Feeding AT and YH caused the increase of jejunal villus height (p < 0.05), and a higher ratio of villus height/crypt depth was observed in AT, BC and YH groups (p < 0.05). The mRNA expression of zonula occludens-1 (ZO-1) in jejunal mucosa was up-regulated by AT, BC and YH diets (p < 0.05). Dietary AT, BC or YH inclusion decreased the interleukin-1β (IL-1β) concentration and TNF-α mRNA expression (p < 0.05), and YH supplementation even down-regulated toll-like receptor 4 (TLR4) and CD14 expressions (p < 0.05). In summary, the dietary administration of BC or YH both improves growth performance through promoting the intestinal barrier function, indicating both of them can serve as potential alternatives to antibiotics growth promoters for the piglet production.     

不同形式补铁补硒对仔猪生长性能、血液生化指标、免疫性能的影响

[目的] 比较母猪和仔猪不同形式的补铁补硒对仔猪生产性能、血液生化指标、免疫性能的影响。[方法] 将32头母猪随机分为8组,每组4头,1~4组基础日粮中添加硫酸亚铁和亚硒酸钠,5~8组基础日粮中添加甘氨酸螯合铁和硒代蛋氨酸。2组和6组母猪所产仔猪于3日龄肌注铁硒合剂;3组和7组仔猪基础日粮中添加硫酸亚铁和亚硒酸钠;4组和8组仔猪基础日粮中添加甘氨酸螯合铁和硒代蛋氨酸;1组和5组仔猪饲喂基础日粮,不接受任何形式的补铁补硒。母猪预产期前30 d开始饲喂试验日粮,仔猪7日龄时开始饲喂试验日粮,试验至仔猪21日龄断奶结束。测定并比较不同组别仔猪生长性能指标、血液生化指标以及免疫性能指标,以及不同组别母猪初乳和常乳中的铁硒含量。[结果] 在仔猪补铁补硒形式相同的条件下,母猪基础日粮中添加甘氨酸螯合铁和硒代蛋氨酸（5~8组）与基础日粮中补充硫酸亚铁和亚硒酸钠（1~4组）相比,能够显著（P<0.05）或极显著（P<0.01）提高仔猪的初生重、断奶重、断奶存活率、断奶合格率,2日龄和21日龄血浆铁含量、血浆硒含量、血红蛋白含量、血浆转铁蛋白含量、IgA含量、IgG含量、IgM含量,以及母猪初乳和常乳中的铁硒含量;在2种形式的母猪补铁补硒条件下,与不补铁补硒相比,仔猪不同补铁补硒形式（2~4组和6~8组）对其断奶重、断奶存活率、断奶合格率,以及21日龄血浆铁含量、血浆硒含量、血红蛋白含量、血浆转铁蛋白含量、谷胱甘肽过氧化物酶活性、IgA含量、IgG含量、IgM含量均有极显著（P<0.01）的提高作用,并且仔猪基础日粮中添加甘氨酸螯合铁和硒代蛋氨酸的效果优于肌注铁硒合剂和基础日粮中补充硫酸亚铁和亚硒酸钠。[结论] 母猪和仔猪基础日粮中添加甘氨酸螯合铁和硒代蛋氨酸,能有效提升仔猪的生长性能、血液生化指标和免疫性能。     

Effect of Se and VE Supplement on Semen Quality,Antioxidant Enzyme Activities and Heat Shock Protein Expression of Goat in High Temperature Season

This experiment was conducted to study the effect of selenium and vitamin E supplement on semen quality, antioxidant enzyme activities and heat shock protein expression of goat in Hainan high temperature season.16 adult Hainan Black goat with good health and approximate weight were randomly divided into 4 groups, fed with basal diet(control group), basal diet+0.5 mg/kg Se(Se group), basal diet+100 mg/kg VE(VE group), and basal diet+0.5 mg/kg Se+100 mg/kg VE(Se+VE group), respectively.The experimental period was 93 d.Semen samples were collected in the last week of the experiment on two consecutive days.The semen quality, antioxidant enzyme activities and heat shock protein expression were analyzed.The results showed that compared with control group, the ejaculate volume was not significantly affected by Se or VE supplement(P>0.05).Sperm density and sperm motility were increased significantly by Se and VE supplement(P<0.05), and the abnormal rate was decreased extremely significantly(P<0.01).The goats fed with Se and VE also had higher activities of GSH-Px(P<0.01), SOD(P<0.05), CAT(P<0.05) and T-AOC(P<0.01), and lower MDA concentration in seminal plasma(P<0.05).The relative expression levels of HSP70 and HSP90 mRNA in supplement groups were decreased extremely significantly(P<0.01).However, there were some certain differences between the Se and VE supplement groups on semen quality and heat shock protein expression.In conclusion, the supplementation of Se and VE could help to improve goat semen quality by increasing the sperm density, sperm motility, the antioxidant enzyme activities, and decreasing the abnormal rate in hot season of Hainan.Finally, Se and VE supplement had good effects on relieving the environment heat stress.     

Construction and Identification of Yeast Surface Display Libraries of Ovis aries DRA Gene Exon 2

The specific primers were designed according to Ovis aries DRA gene sequence deposited in GenBank and the multiple cloning site of the plasmid pYD1,which was a vector used for protein surface display on Saccharomyces cerevisiae.The gene encoding DRA was amplified by PCR using the genomic RNA of Ovis aries.The 762 bp fragment was cloned and released in GenBank and registration number was KR422362.The PCR product was inserted into the yeast surface display plasmid vector pYD1 by double enzyme digestion.It was indicated that DRA gene was successfully integrated into the genome.Dot mutation was made at both ends of exon 2 in DRA gene for making restriction enzyme cutting site and design the exon 2 specific primers according to mutated Ovis aries DRA gene sequence.Sequenced exon 2 amplification products based on DNA pooling of sheep large sample template was analyzed the polymorphic loci.The polymorphic exon 2 246 bp fragment was obtained by double enzyme digestion and connected to surface display restructuring mutation carriers pYD1-DRA by the same double enzyme digestion,and then we successfully constructed yeast surface display libraries.We transformed it into Saccharomyces cerevisiae EBY100 cell.Yeast monoclone was identified by PCR amplification and sequencing,and we confirmed that DRA gene had been integrated into Saccharomyces cerevisiae genome.After galactose induced,it was detected that DRA gene library had been successfully demonstrated on the yeast cell surface under the fluorescence microscope by immunofluorescence method.     

葡萄VvSUC12基因启动子的克隆及上游调控因子鉴定

枝干病害葡萄溃疡病近年来严重限制了葡萄产业发展。研究表明葡萄蔗糖转运蛋白参与寄主植物和病原菌的互作过程。为解析蔗糖转运蛋白VvSUC12在葡萄免疫反应过程中的功能,本研究克隆了VvSUC12基因翻译起始位点上游1 500 bp的启动子区,通过生物信息学分析发现该区域包含4个Dof转录因子结合序列(A/TAAAG)及多种激素调节与防御相关的顺式元件。实时荧光定量分析显示,接种可可毛色二孢菌显著诱导VvSUC12和VvDof19基因的表达。通过酵母单杂交实验筛选得到与VvSUC12启动子互作的转录因子VvDof19。酵母双杂交实验证实VvDof19具有自激活活性;进一步通过烟草瞬时表达发现Dof19-GFP的相对GUS活性约为对照GFP的9倍,表明转录因子VvDof19能够激活VvSUC12基因的表达。研究结果为深入研究蔗糖转运蛋白在葡萄免疫反应中的功能奠定基础。     

石墨炉原子吸收光谱法测定新资源食品中的硒含量

建立一种简便、快速、准确的食品中硒的测定方法;检测新资源食品芝麻叶、野生四叶参、野生菜葛根、野生葛花及桑蚕蛹中的硒含量。用1mg·mL-1硝酸镍为基体改进剂,加入体积分数为0.1%TritonX-100和0.5%硝酸(1∶1),以改善样液的表面张力。用HITACHIZ-5000型石墨炉原子吸收光谱仪及全自动进样方法进行硒含量测定。石墨炉采用光控升温方式,硒的灰化温度按程序升温方式进行,程序升温分为干燥、灰化、原子化和净化。该方法的测定限为18.6μg/L,精密度为3.26%￣9.29%(RSD),加标回收率为92.1%￣108.8%。该方法具有测定简便、快速和准确等特点,可作为食品中硒含量测定的良好方法。     

酵母细胞壁多糖与铝硅酸盐复合物对猪生长性能、免疫指标及养分消化率的影响

本研究旨在探讨酵母细胞壁多糖(yeast polysaccharide,YP)与铝硅酸盐复合物(alumi-nosilicate complex,AC)对猪的生长性能、免疫指标及养分消化率的影响。试验选用平均体重为(66.18±1.26)kg的"杜×长×大"三元杂交猪140头,随机分成5组,每组4个重复,每个重复7头猪。各组分别饲喂添加不同剂量复合物的试验饲粮,A组为对照组,不添加复合物;B组添加0.1%YP;C组添加0.1%YP和0.9%AC;D组添加0.1%YP和1.9%AC;E组添加0.1%YP和2.9%AC。试验期51 d。结果表明:1)D、E组平均日增重显著低于对照组(P<0.05),各组平均日采食量和料重比差异不显著(P>0.05);2)与对照组相比,D组血清谷胱甘肽过氧化物酶(GSH-Px)活性及补体3(C3)、免疫球蛋白M(IgM)、免疫球蛋白A(IgA)、免疫球蛋白G(IgG)水平显著提高(P<0.05),血清超氧化物歧化酶(SOD)活性和补体4(C4)水平无显著差异(P>0.05);3)各组养分消化率无显著差异(P>0.05)。由此可见,YP和AC复合使用降低了育肥猪的生长性能,0.1%YP和1.9%AC同时添加可增强生长猪的免疫功能。     

酵母壁多糖对断奶仔猪肠道挥发性脂肪酸和微生物菌群的影响

本研究旨在探讨饲粮中添加酵母壁多糖对断奶仔猪肠道挥发性脂肪酸和微生物菌群的影响。试验采用单因素试验设计方法,选取180头遗传背景一致、健康状况良好、胎次和体重接近的21日龄断奶仔猪,随机分为4个组,每组5个重复,每个重复9头猪。4个组试验猪分别饲喂对照饲粮(未添加酵母壁多糖)、0.15%酵母壁多糖饲粮、0.30%酵母壁多糖饲粮和0.45%酵母壁多糖饲粮。试验期21 d。结果表明:1)与对照组相比,饲粮中添加0.15%、0.30%和0.45%酵母壁多糖显著提高了仔猪结肠乙酸的含量(P0.05);其中,0.30%和0.45%酵母壁多糖还显著提高了结肠丙酸、丁酸以及总挥发性脂肪酸的含量(P0.05),且二者之间差异不显著(P0.05)。2)与对照组相比,饲粮中添加0.15%、0.30%和0.45%酵母壁多糖显著降低了盲肠沙门氏菌和大肠杆菌数量(P0.05),且0.30%和0.45%酵母壁多糖组之间差异不显著(P0.05)。由此可知,酵母壁多糖可提高仔猪肠道挥发性脂肪酸含量,并改善肠道微生物菌群结构,根据回归方程预测,酵母壁多糖在仔猪饲粮中的适宜添加水平为0.31%~0.40%。     

Screening and Analysis of Proteins Interacting with TaPDK from Physiological Male Sterility Induced by CHA in Wheat

To further research the regulatory network of pyruvate dehydrogenase kinase (designated as TaPDK) in physiological male-sterility (PHYMS) of wheat induced by chemical hybridizing agent (CHA) SQ-1, an anther cDNA library was constructed, and the proteins interacting with TaPDK were screened via yeast two-hybrid technique. Subsequently, a few candidate proteins in nucleotide expression levels were detected by real-time quantitative PCR. Yeast-two hybrid screening was performed by mating yeast strain Y2HGold containing BD-TaPDK bait plasmid with yeast strain Y187 including anther cDNA library plasmid. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Ade/-His/-Leu/-Trp) (QDO), and further were incubated on QDO medium containing AbA and X-α-Gal. The interactions between TaPDK and the proteins obtained from positive colonies were further confirmed by co-transformation validation. After plasmids DNA were extracted from blue colonies and sequenced, the sequences results were analyzed by bioinformatic methods. Finally, 24 colonies were obtained, including eight genes, namely non-specific lipid-transfer protein precursor (TanLTP), polyubiquitin (TaPUbi), glyceraldehyde-3-phosphate dehydrogenase, proliferating cell nuclear antigen (TaPCNA), CBS domain containing protein (TaCBS), actin, guanine nucleotide-binding protein beta subunit, chalcone synthase, and three new genes with unknown function. The results of quantitative RT-PCR showed that the expression levels of TanLTP, TaPUbi, and TaPCNA were obviously up-regulated in PHYMS anther, and TaCBS expression was only increased at the tricellular stage in PHYMS anther compared with in fertile lines. Whereas, the expression of TaPDK was obviously down-regulated in PHYMS lines. Collectively, these datas indicated that the majority of candidate proteins might be related to pollen abortion in PHYMS lines, which further suggested that TaPDK plays multiple roles in pollen development, besides participating in regulating pyruvate dehydrogenase complex activity.     

2种玉米青贮饲料青贮过程中主要微生物的变化规律研究

旨在研究玉米青贮过程中各主要微生物的数量变化及变化趋势.做去棒揉丝和带穗切段瓶制青贮,取青贮前玉米青贮原料及在装瓶后第0.5、1、2、3、4、5、6、7、9、11、13、15、20、25、30、35、40、45、50天的青贮样品,用无菌水浸泡,然后用选择性培养基培养其中的主要微生物,包括乳酸菌、酵母菌和霉菌,最后进行计数.结果表明,在玉米青贮密封后60 d内,水分含量变化很小.pH在第2天下降到4以下,然后稳定在3.5左右,带穗切段玉米青贮pH低于去棒揉丝玉米青贮.乳酸菌数量剧增,在第6、7天时达到最高峰,为109数量级,之后数量缓慢下降,于第15～20天时稳定在107数量级,去棒揉丝玉米青贮的乳酸菌数量在第7天达到最高峰,带穗切段玉米青贮则在第6天出现最高峰,其稍高于去棒揉丝玉米青贮.酵母菌在青贮初期数量有些波动,出现最高峰到达107数量级后数量随时间的延长而减少,去棒揉丝玉米青贮时酵母菌数量迅速下降,第40天后没有再检测到酵母菌的存在,而带穗切段玉米在青贮12 h之内数量迅速增加,之后缓慢下降,第50天后没有再检测到酵母菌的存在.霉菌因密封后氧气缺乏而数量很快减少,最晚在第11天左右便检测不到,去棒揉丝玉米青贮中霉菌数量下降较迅速,第4天之后便检测不到霉菌存在,而带穗切段玉米青贮则比较平缓,直到第11天之后便检测不到霉菌存在.综上所述,在玉米青贮过程中主要微生物随时间大致呈现逐步减少的趋势；对玉米秸秆进行纵切揉丝处理有利于玉米青贮饲料的制作.