菌丝霉素分泌型丁酸梭菌防治罗非鱼链球菌病的应用研究

为解决罗非鱼链球菌感染难题,本研究在抗菌肽菌丝霉素分泌型丁酸梭菌制剂的基础上,对其进行了抑菌活性、稳定性和罗非鱼养殖应用研究。结果发现:表达产物菌丝霉素对常见致病菌如大肠杆菌、沙门氏菌、金黄色葡萄球菌、蜡样芽孢杆菌和猪链球菌均有较好的抑菌活性。选用米糠作为菌丝霉素分泌型丁酸梭菌的保护剂载体,此制剂可在常温保存一年。通过上述的应用基础试验,初步建立一套菌丝霉素分泌型丁酸梭菌制剂养殖应用方案,此方案可显著降低罗非鱼链球菌病发病率58.6%以上。本研究为丁酸梭菌制剂在防治罗非鱼链球菌病应用方面提供了参考,此制剂有望替代抗生素,促进罗非鱼养殖业的持续健康发展。     

发酵构树饲料对断奶仔猪生长性能、养分表观消化率和血清生化指标的影响

本试验旨在研究饲粮中不同比例发酵构树饲料对断奶仔猪生长性能、养分表观消化率和血清生化指标的影响。试验选取35日龄、体重为(7.4±1.1)kg的健康断奶仔猪216头,随机分为Ⅰ(对照)、Ⅱ、Ⅲ、Ⅳ组,每组3个重复,每个重复18头仔猪。其中,Ⅰ组饲喂基础饲粮;Ⅱ组饲喂3%发酵构树饲料+97%基础饲粮;Ⅲ组饲喂6%发酵构树饲料+94%基础饲粮;Ⅳ组饲喂9%发酵构树饲料+91%基础饲粮。预试期5 d,正试期25 d。结果表明:1)与Ⅰ组相比,Ⅱ、Ⅲ、Ⅳ组断奶仔猪的腹泻率显著降低(P<0.05),各组间平均日采食量(ADFI)、平均日增重(ADG)和料重比(F/G)无显著差异(P>0.05)。2)与Ⅰ组相比,Ⅱ、Ⅲ、Ⅳ组粗蛋白质(CP)、粗脂肪(EE)、钙(Ca)和磷(P)表观消化率均显著降低(P<0.05)。3)Ⅲ组血清总胆固醇(TC)含量较Ⅰ、Ⅱ、Ⅳ组显著降低(P<0.05);各组间其他血清生化指标无显著差异(P>0.05)。4)各组间血清超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量及总抗氧化能力(T-AOC)无显著差异(P>0.05)。综上所述,饲粮中不同比例发酵构树饲料可以显著降低断奶仔猪的腹泻率,未对生长性能、血清生化指标和抗氧化指标产生不利影响,其中6%的替代比例效果最佳。     

凉茶渣的黑曲霉发酵工艺和抗氧化活性研究

为促进凉茶渣在畜牧业中的资源化利用,本研究以黑曲霉为菌种固态发酵凉茶渣,首先在单因素试验条件下考察了时间、温度、含水量、浸泡液pH、氮源和碳源对凉茶渣降解率和产物pH的影响;再根据单因素试验结果和规模化生产实际需要,以4%硫酸铵为氮源,2%葡萄糖为碳源,以降解率为考察指标,通过正交试验优化发酵工艺;并通过检测超氧自由基清除率、羟基自由基清除率和1,1-二苯基-2-三硝基苯肼（DPPH）自由基清除率,评价凉茶渣在最优工艺条件下发酵前后抗氧化活性的变化。结果发现,含水量为60%,浸泡液pH为9.0,31℃发酵168 h是凉茶渣的最佳发酵工艺参数。最佳工艺条件下凉茶渣的降解率为25.23%,发酵产物pH为4.53。当凉茶渣发酵前水提液浓度为24 mg/mL时,超氧自由基清除率为43.56%,羟基自由基清除率为47.06%,DPPH自由基清除率为90.71%;当最优条件下发酵产物水提液浓度24 mg/mL时,超氧自由基清除率为30.77%,羟基自由基清除率为95.63%,DPPH自由基清除率为87.36%。本试验结果表明,黑曲霉是适宜的凉茶渣发酵菌种,且凉茶渣经过黑曲霉发酵后具有良好的抗氧化活性。     

猪流行性腹泻病毒S1D蛋白的优化表达及其单克隆抗体的制备

为进一步探究猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)S蛋白的抗原表位及其功能,本试验通过优化S1D基因的密码子,构建了S1D基因未优化的重组原核表达质粒pET-S1D和已优化的pET-ΔS1D,并进行了诱导表达和纯化。使用SDS-PAGE和Western blotting方法验证S1D、ΔS1D蛋白在大肠杆菌内得到正确表达,利用Image J软件对S1D、ΔS1D蛋白表达量进行灰度扫描,通过t检验分析两者差异性。将纯化的ΔS1D 蛋白免疫 BALB/c小鼠,通过细胞融合、筛选及亚克隆,获得单克隆细胞株。利用体内诱生法制备抗PEDV S1D蛋白的单克隆抗体腹水,使用ELISA、Western blotting、间接免疫荧光试验3种方法对腹水效价及特异性进行检测和验证。SDS-PAGE和Western blotting结果显示,表达S1D、ΔS1D蛋白的样品均在34 ku处出现正确的目的条带。t检验结果表明, S1D、ΔS1D两者蛋白表达量差异极显著(P<0.01)。ELISA结果显示, 腹水的抗体效价达到了1∶1 000 000,腹水与PEDV病毒粒子和纯化后的ΔS1D蛋白反应均呈阳性,与PEDV N蛋白、pET-32a(+)空载体蛋白和猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)、猪传染性胃肠炎病毒(Transmissible gastroenteritis virus,TGEV)、猪瘟病毒(Classical swine fever virus,CSFV)、猪德尔塔冠状病毒(Porcine deltacoronavirus,PDCoV)和猪急性腹泻综合征冠状病毒(Swine acute diarrhea syndrome coronavirus,SADS-CoV) 5种病毒反应均呈阴性。Western blotting结果显示,腹水与ΔS1D蛋白和PEDV S蛋白分别在34和180 ku处有特异性条带出现,与pET-32a(+)空载体蛋白、正常Vero细胞蛋白均无特异性条带出现。间接免疫荧光试验结果显示,腹水及阳性对照组均能使细胞出现特异性绿色荧光信号,而空白及阴性对照组均未见绿色荧光信号。密码子优化可在原核表达系统中显著提高重组蛋白的表达水平,本研究基于高效表达的ΔS1D蛋白,成功制备了1株能稳定分泌与 PEDV S蛋白特异性结合的单克隆抗体的细胞株,为进一步探究 PEDV S蛋白抗原表位及蛋白功能的研究奠定了基础。     

Effects of dietary supplementation of nucleotides from late gestation to lactation on the performance and oxidative stress status of sows and their offspring

Increased metabolic burdens in breeding sows, which are induced by elevated systemic oxidative stress, could increase the need for nucleotides to repair lymphocyte DNA damage; however, de novo synthesis of nucleotides may be insufficient to cover this increased need. This study investigated the effects of dietary nucleotides on milk composition, oxidative stress status, and the reproductive and lactational performance of sows. Forty multiparous sows were assigned to 2 dietary treatments (Control group, and 1 g/kg Nucleotides group) based on a randomized complete block design using their BW at 85 d of gestation as a block. Sows from 2 groups were fed a restricted diet during gestation and ad libitum during lactation. The experiment lasted from 85 d of gestation to 21 d of lactation. The reproductive performance of sows and the growth performance of suckling piglets were measured. Oxidative stress parameters and milk components were also analysed. Data were analyzed using contrasts in the MIXED procedure of SAS. Sows in the Nucleotides group consumed more feed during the first week (P < 0.01) and from 1 to 21 d (P < 0.05) of lactation than those in Control group. Correspondingly, the litter weight gain of piglets showed a tendency to increase from cross-fostering to 9 d (P = 0.09) and from cross-fostering to 20 d (P = 0.10) in the Nucleotides group relative to the Control group. Additionally, the Nucleotides group was higher (P < 0.01) than the Control group in the concentrations of uridine 5''monophosphate, guanosine 5''monophosphate, inosine 5''monophosphate, adenosine 5''monophosphate and total nucleotides in milk. Furthermore, the Nucleotides group was higher (P < 0.01) than the Control group in the serum levels of total antioxidant capacity (P < 0.01) for sows at 109 d of gestation and glutathione peroxidase for weaning piglets, but lower at the levels of thiobarbituric acid-reactive substances (P < 0.05) in serum of weaning piglets. This study indicated that maternal dietary nucleotides could promote piglet growth, probably due to the higher lactational feed intake and higher concentration of nucleotides in the milk of sows, and lower oxidative stress for both sows and piglets.     

从女性主义视角看《包法利夫人》

从18世纪到20世纪60年代,自由平等思潮揭开了轰轰烈烈的女性运动序幕,女性的个人权利和政治、宗教信仰自由,女性对于生命、自由、追求幸福的选择权以及自我决定权开始广泛受到关注。《包法利夫人》在一定程度上体现了当时女性主义的进步性,同时也揭示出在父权制主流社会下女性(书中的女主人公)注定要走向堕落的悲剧宿命。     

Seasonal and photoperiodic regulation of secretion of hormones associated with reproduction in Magang goose ganders

This study examined the reproductive endocrine profile under natural and artificial photoperiods in Magang goose ganders. Group 1 ganders (n=8) served as non-treated controls and were exposed to natural photoperiod throughout the experiment from 13th January to 17th December 2004. Group 2 ganders (n=8) were exposed to 18 h long daily photoperiod for 60 days from 13 January till 15 March 2004 and again to 16 h photoperiod for 75 days till 10th October 2004, and the 11h short photoperiod in the remainder periods of the experiment. In control ganders, plasma LH concentrations were high in normal breeding seasons (August-March) and decreased to low levels in non-breeding season from April to July. Testosterone concentrations changed similarly to that of LH throughout the seasons. Seasonal pattern of PRL concentrations was opposite to those of LH and testosterone, with low values in breeding season and high values in non-breeding season. In artificial photoperiod treated ganders, increasing photoperiod increased PRL and decreased LH and testosterone concentrations, while decreasing photoperiod reversed these changes. There were no seasonal or photoperiod caused changes in plasma T3 concentrations in both control ganders and artificial photoperiod treated ganders. These results demonstrated that in Magang goose ganders that long photoperiod stimulates PRL secretion and decreases LH secretion, which terminates reproductive season in spring and early summer, and short photoperiod stimulates LH secretion and inhibits PRL secretion rendering ganders enter into reproductive season.     

An 8bp indel in exon 1 of Ghrelin gene associated with chicken growth

Ghrelin, acts as the endogenous ligand for growth hormone secretagogues receptor (GHS-R), is a novel growth hormone (GH) releasing peptide with reported effects on food intake in chickens. In this study, an 8 bp indel polymorphism in exon 1 of the chicken Ghrelin (cGHRL) gene was genotyped in a F(2) designed full-sib population to analyze its associations with chicken growth and carcass traits. Later, mRNA level in the proventriculus was determined by real-time PCR to reveal the expression feature of cGHRL gene. Result showed that this 8 bp indel was significantly associated with body weight at the age of 28 days (BW28) and 56 days (BW56), eviscerated weight (EW) and leg muscle weight (LMW) (P<0.05), highly significantly associated with hatch weight (HW), BW14, 21, 35, 42, 49, 90 and body length (BL), dressed weight (DW), eviscerated weight with giblet (EWG), wing weight (WW), breast muscle weight (BMW) and head and neck weight (HNW) (P<0.01). Meanwhile, A allele (with 'CTAACCTG') was positive for chicken growth as individuals with AA genotype had the highest value of all traits. Analysis on cGhrelin mRNA level revealed that it differed significantly among individuals with three genotypes (P<0.05). Individuals with AB genotype had the highest mRNA level, whereas that of AA had the lowest one. It was concluded that this 8 bp indel of cGHRL gene was significantly associated with most body weight and body composition traits, and negative effect of endogenous Ghrelin on chicken growth were indicated by this study.     

沙对高尔夫果岭坪床物理特性的影响

对28种沙与泥炭按75∶25混配而成的高尔夫果岭坪床材料的物理特性进行了评价。不同类型沙的粒径、均一性和形状有所不同。高尔夫果岭坪床的物理特性主要取决于所使用的沙的特性。对沙 泥炭混配的坪床来说,其导水率在95~827 mm/h范围内变化,总孔隙度变化范围为38.8%~47.2%,而通气孔隙度(在-3 kPa水势下)为0.9%~24.6%,毛管孔隙度(在-3 kPa水势下)为17.7%~42.1%。结果表明,排水速率和通气孔隙度与毛管孔隙度的平衡主要受沙的粒径大小的影响,起决定作用的是粒径最大和最小的沙所占的比例,而容重、总孔隙度和抗剪切强度取决于沙的均一性。当沙粒较圆且较为光滑时,坪床材料孔隙度趋向减少。沙表面越粗糙,坪床抗剪切能力就越强,而沙对坪床硬度的作用不显著。     

基于最大熵模型预测小火蚁在我国的适生区

小火蚁是中国大陆新发现的重要外来入侵害虫, 目前对该虫的传入来源和在我国的适生区范围尚不明确?为明确我国小火蚁适生区范围, 有效防控该虫在我国的扩散和蔓延, 本研究通过该虫全球已有的分布数据, 采用最大熵模型对其适生区进行了预测?研究表明, 最大熵模型预测小火蚁适生区精度较高?预测结果显示, 我国小火蚁的潜在适生区主要分布于南方, 其中, 高适生区分布在台湾?海南?云南南部边境?广西西南局部?福建西南部?广东南部及其沿海地区?预测结果与该物种现有地理范围的生态条件一致?年降水量对小火蚁的适生性影响最大, 理论年降水量为2 040 mm时小火蚁分布的概率最高?随着全球气候变暖, 未来我国小火蚁的适生区有向北扩大的趋势, 但主要适生区还是以南方为主?