禁食禁水对新疆伊犁马生化指标及应激相关激素的影响

为探究禁食禁水双重应激对马匹机体的影响,试验选取8匹成年伊犁骟马,采用不完全拉丁方试验设计,分为禁食禁水组(禁食禁水49 h)和正常饲喂组(对照组),在试验过程中监测马匹基本生命体征、血液生化指标及应激相关激素含量的变化。结果表明,禁食禁水组心率在12~48 h均显著低于对照组(P< 0.05),呼吸频率在48和49 h显著低于对照组(P< 0.05),血糖浓度在18、24和36 h均显著低于对照组(P< 0.05),血浆皮质醇水平在18、30和36 h均显著高于对照组(P< 0.05),血浆抗利尿激素水平在48和49 h均显著高于对照组(P< 0.05),其他指标未见显著变化(P> 0.05)。由此可知,在禁食禁水双重应激情况下,马匹通过降低心血管活动、减少机体能量消耗、提高应激能力来维持基本生命活动。     

运输后禁食与非禁食对肉兔应激水平、肉品质和行为的影响

本试验以伊拉肉兔为研究对象,探究肉兔运输后在禁食与非禁食条件下对其应激水平、肉品质和行为的影响,探索合理的肉兔运输后的处理方式。试验选取80只体重相近（（2.5±0.5） kg）的伊拉公兔,随机分为8个组：对照组、运输后0 h组、非禁食6 h组、非禁食24 h、禁食6 h组、禁食24 h组、禁食行为观察组和非禁食行为观察组（做行为观察,不屠宰）。预试期为7 d,正式期的运输时间为2 h。结果发现,在血清生化测定中,和对照组相比,运输后0 h组血清ALT、AST、TG、Glu、CK、LDH水平无显著变化（P>0.05）,而运输后0 h组血清TP浓度显著下降,皮质醇浓度显著上升（P<0.05）。非禁食24 h组血糖水平高于禁食24 h组（P<0.05）、血清CK浓度低于禁食24 h组（P<0.01）,非禁食6 h的LDH水平低于禁食6 h组（P<0.05）。在肉品质测定中,运输后0 h时肌肉pH显著上升（P<0.05）,肌肉L^*、a^*水平显著下降（P<0.05）,而非禁食24 h组的L^*、a^*和b^*值显著高于禁食24 h组。而禁食组与非禁食组的肌肉pH、失水率、蒸煮损失率和剪切力等无显著差异（P>0.05）。在基因表达方面,运输后0 h时JNK、Caspase-3的mRNA水平显著上升（P<0.05）。非禁食相比于禁食降低了JNK mRNA相对表达量（P<0.05）,对Caspase-3水平无显著影响（P>0.05）。在行为学方面,非禁食相比于禁食降低了肉兔的食粪行为（P<0.05）,对于肉兔刻板行为改变不明显（P>0.05）。本研究提示,在运输后静养期间,24 h非禁食能一定程度降低肉兔的应激、减少机体损伤、改善兔肉品质,有利于提高肉兔的福利水平。     

SIRT1对牦牛卵母细胞体外成熟与老化的影响

旨在探索SIRT1在牦牛卵母细胞体外成熟与老化过程中的作用。本研究在体外成熟液中分别添加SIRT1特异性激动剂SRT2104（SRT组）和特异性抑制剂Inauhzin（INZ组），牦牛卵丘卵母细胞复合体（COCs）体外培养24 h后，观察卵丘细胞的扩展和第一极体的排出情况；利用免疫荧光检测体外培养24与36 h后卵母细胞内的ROS水平；采用实时荧光定量PCR法检测体外培养24与36 h后卵母细胞内SIRT1、FOXO3a、SOD2以及Bax的表达水平；体外培养24与36 h后的牦牛卵母细胞进行体外受精，观察并统计其卵裂率与囊胚形成率。结果显示，体外培养24 h，SRT组的卵丘细胞扩展程度显著高于对照组（P<0.05），而INZ组的卵丘细胞扩展程度和第一极体排出率显著低于对照组（P<0.05）。随着体外培养时间的增加，卵母细胞内的ROS水平显著增加（P<0.05）；添加SRT2104能显著抑制卵母细胞中ROS水平的积累（P<0.05），而添加Inauhzin则显著上调卵母细胞内的ROS水平（P<0.05）。体外培养24 h后，SRT组SIRT1、FOXO3a与SOD2的表达水平显著高于对照组（P < 0.05），但Bax的表达水平显著降低（P<0.05）；INZ组的SIRT1、FOXO3a与SOD2表达均显著低于对照组（P<0.05），但Bax的表达水平显著上调（P<0.05）。牦牛卵母细胞体外培养24 h后，SRT组的卵裂率与囊胚形成率显著高于INZ组和对照组（P<0.05）；卵母细胞体外培养36 h后，INZ组的卵裂率和囊胚形成率显著低于其他组（P<0.05）。综上表明，SIRT1参与了牦牛卵母细胞的体外成熟，在体外培养液中适当添加SIRT1激动剂，有利于卵母细胞体外成熟及缓解老化，同时改善早期胚胎的发育能力。     

伪狂犬病毒感染对BHK-21悬浮细胞内质网应激和未折叠蛋白反应的影响

【目的】探讨伪狂犬病毒(PRV)感染对BHK-21悬浮细胞内质网应激及未折叠蛋白反应(UPR)的影响。【方法】将悬浮的乳仓鼠肾细胞(BHK-21)以感染复数(MOI)0.01接种PRV,分别在接毒后12、24、36、48和56 h取样,用CCK-8法检测细胞存活率,按Reed-Muench法检测病毒滴度,筛选病毒接种处理的合适时间;以不接毒细胞作对照组,分别在感染后12、24、36和48 h取样,用实时荧光定量PCR法检测内质网应激标志分子葡萄糖调节蛋白78(GRP78)及UPR的3种感受蛋白(PERK、IRE1、ATF6)相关通路中基因的表达变化,Western blotting法检测相关蛋白的表达变化。在接种PRV同时分别加入0、0.001、0.005、0.01和0.02 μmol/L内质网应激诱导剂毒胡萝卜素(Tg)和0、20、40、80和160 μmol/L内质网应激抑制剂牛磺熊去氧胆酸(TUDCA),48 h收集细胞测定病毒滴度和细胞存活率。【结果】PRV感染56 h细胞存活率低于70%,感染48 h时病毒滴度达到最高(8.1 lg TCID₅₀/mL),因此后续试验分别在接毒后12、24、36和48 h取样。与对照组相比,PRV感染36和48 h GRP78转录水平均极显著升高(P＜0.01);PERK通路中,转录激活因子4(ATF4)转录水平在PRV感染36、48 h均极显著升高(P＜0.01),生长抑制DNA损伤基因34(GADD34)转录水平在PRV感染36 h显著升高(P＜0.05)、48 h极显著升高(P＜0.01),磷酸化真核翻译起始因子(P-eIF2α)蛋白水平在PRV感染36、48 h均极显著升高(P＜0.01);IRE1通路中,PRV感染组细胞36 h后多以剪切形式sXBP1存在,同时下游p58^IPKmRNA水平在36 h显著增加(P＜0.05),p58^IPK和EDEM mRNA水平在48 h均极显著增加(P＜0.01);ATF6通路中,PRV感染不同时间ERp57、PDI、Calnexin和Calreticulin的表达均无显著变化(P＞0.05)。与0 μmol/L组相比,0.01和0.02 μmol/L Tg极显著降低细胞存活率(P＜0.01),0.005和0.01 μmol/L Tg均极显著增加了PRV病毒滴度(P＜0.01)。【结论】PRV感染可以诱导BHK-21悬浮细胞内质网应激,激活UPR的PERK和IRE1信号通路,说明PRV利用内质网应激增强其复制。     

Effects of Supplemental Oil on the Sport Performance,Plasma Antioxidant Capacity and other Metabolic Index of Yanqi Horse

This test mainly studied the effects of supplemental different levels of oil on the performance, plasma antioxidant capacity and other metabolic index of Yanqi horse.9 Yanqi horses were randomly divided into three groups according to performance, body weight, body measurement, age.The control group was fed with basal diet.Horses in the groups Ⅰ and Ⅱ were fed with diet supplemented with 2.5% and 5% of oil.All horses were participated in 20 km racing, on the 20th day.Temperature, heart rate, respiratory rate of all horses were measured, 30 min after the racing.Simultaneously, blood samples were collected from the jugular vein.The results showed that the heart rate value of groups Ⅰ and Ⅱ were highly significant lower than control group (P<0.01), 0 min after the racing.The group Ⅱ was highly significantly lower than control group (P<0.01), 5 min after the racing.The plasma GSH-Px and T-SOD activity of groups Ⅰ and Ⅱ compared with the control group respectively increased by 87.60%(P<0.05), 15.87%(P>0.05) and 98.38%(P<0.05), 38.25%(P<0.05);The plasma MDA and LA concentration of groups Ⅰ and Ⅱ compared with the control group respectively decreased by 53.44% (P<0.05), 24.08%(P<0.05) and 51.42% (P<0.05), 25.08%(P<0.05).In conclusion, the performance and antioxidant capacity of Yanqi horse could be improved by the basal diet added different levels of oil.The physiological function of Yanqi horse could be fast recovery after the racing.Compared, added 5% oil feed supplement was especially effective.     

添加油脂对焉耆马运动性能、血浆抗氧化能力及代谢指标的影响

试验旨在探讨日粮中添加油脂对焉耆马赛后运动性能及抗氧化能力的影响。选取运动成绩、体重、体尺、年龄相近的焉耆马9匹,随机分为3组,对照组饲喂基础日粮,试验Ⅰ、Ⅱ组分别在基础日粮中添加不同水平的豆油。试验开始第20天对各组马匹进行20 km模拟比赛,赛后30 min内测定各组试验马匹的体温、脉搏、呼吸,同时采集各组马匹血液。结果表明,试验Ⅰ、Ⅱ组焉耆马赛后0 min心率极显著低于对照组(P<0.01),试验Ⅱ组焉耆马赛后5 min呼吸频率极显著低于对照组(P<0.01);试验Ⅰ、Ⅱ组焉耆马赛后血浆中GSH-Px、T-SOD活力浓度分别比对照组高87.60%(P<0.05)、15.87%(P>0.05)和98.38%(P<0.05)、38.25%(P<0.05);试验Ⅰ、Ⅱ组焉耆马血浆中MDA、LA浓度分别比对照组低53.44%(P<0.05)、24.08%(P<0.05)和51.42%(P<0.05)、25.08%(P<0.05)。饲料中添加油脂可提高焉耆马运动性能,有利于焉耆马运动后生理机能的快速恢复,提高运动后焉耆马的抗氧化能力,相比较而言,添加5%油脂组效果较佳。     

添加不同水平单宁酸对绵羊体外发酵参数和甲烷产量的影响

本试验以精料、青干草和单宁酸作为发酵底物,研究添加不同水平单宁酸对绵羊体外发酵参数和甲烷产量、干物质和粗蛋白质降解率的影响,以期为体内试验筛选适宜的单宁酸浓度,为富含单宁的粗饲料资源应用于反刍动物日粮配制及减少反刍动物养殖过程种甲烷排放提供科学依据。采用体外批次培养法开展研究,发酵底物为精料、青干草和单宁酸,设置底物的精粗比为3∶7。根据单宁酸添加量分为6组,单宁酸添加量分别为0、0.5%、1.0%、1.5%、2.0%、3.0%,发酵时间设置为3、6、12和24 h,每个处理3个重复,测定各培养时间pH、氨氮（NH₃-N）浓度、挥发性脂肪酸（VFA）浓度、总产气量、甲烷产量、干物质和粗蛋白质降解率。结果显示,培养至3 h时,1.0%和2.0%单宁酸添加组pH显著高于对照组（P<0.05）,其他时间点各组pH无显著差异（P>0.05）;1.0%、2.0%和3.0%单宁酸添加组NH₃-N浓度显著低于对照组（P<0.05）;培养至6和24 h时,各处理组NH₃-N浓度均显著低于对照组（P<0.05）。培养至6 h时,2.0%单宁酸添加组乙酸、丙酸、丁酸和总VFA浓度均显著低于对照组（P<0.05）;培养至24 h时,1.0%、1.5%和3.0%单宁酸添加组的丙酸、丁酸和总VFA浓度均显著低于对照组（P<0.05）;各单宁酸添加组4个时间点丙酸、丁酸、总VFA浓度的平均值均显著低于对照组（P<0.05）。培养至12、24 h时,各处理组总产气量与对照组差异不显著（P>0.05）;培养至12 h时,2.0%组甲烷产量显著低于对照组、0.5%组和1.0%组（P<0.05）;培养至24 h时,2.0%和3.0%组的甲烷产量显著低于对照组（P<0.05）。因此,添加单宁酸会显著降低体外发酵的NH₃-N浓度,抑制VFA的产量,添加1.5%、2.0%的单宁酸能够显著降低瘤胃发酵甲烷产量,添加单宁酸的处理组粗蛋白质降解率显著低于对照组。     

光照周期对褪黑激素受体在母兔下丘脑-垂体-卵巢轴中分布与表达的影响

旨在观察不同光照周期对新西兰白色母兔褪黑激素受体在"下丘脑-垂体-性腺轴"中的分布和表达模式,从而进一步分析褪黑激素在不同光照周期下调控母兔发情的机制。选用5月龄未经产新西兰母兔48只,随机分成3组,每组16只,前10 d各试验组采用"12 h光照：12 h黑暗"的光照制度,后6 d分别采用长光照（16 h光照：8 h黑暗）、短光照（8 h光照：16 h黑暗）和正常光照（12 h光照：12 h黑暗,对照组）的光照制度,光照强度80 lx,试验期为16 d。试验结束后剖杀动物,取下丘脑、垂体和卵巢,用qPCR、免疫印迹、免疫组织化学方法,研究不同光照周期对母兔的下丘脑、垂体、卵巢中褪黑激素受体亚型分布与表达的影响。结果表明：在下丘脑,短光照组的MT1 mRNA表达量较长光照组高88.8%（P<0.05）,较对照组高54.9%（P<0.05）,长光照组与对照组间无显著差异;MT2 mRNA表达量在不同光周期组间差异不显著（P>0.05）。免疫组织化学染色结果显示,长光照组下丘脑室周核的MT1阳性细胞数明显较短光照组少69.4%（P<0.05）,较对照组少37.3%（P<0.05）,短光照组比对照组显著多104.8%（P<0.05）;同样,长光照组室旁核的MT1阳性细胞数明显较短光照组少52.9%（P<0.05）,对照组明显较短光照组少55.8%（P<0.05）,而长光照组与对照组间差异不显著（P>0.05）。在垂体,短光照组的MT1 mRNA表达量比长光照组显著高164%（P<0.05）,比对照组显著高49.5%（P<0.05）,而长光照组比对照组显著低43.5%（P<0.05）;但不同光周期下MT2 mRNA表达量差异不显著（P>0.05）;长光照组的卵巢MT1 mRNA表达量较对照组低33.3%（P<0.05）,较短光照组低53.6%（P<0.05）,短光照组与对照组间差异不显著（P>0.05）;卵巢中短光照组MT2的mRNA相对表达量比对照组显著高90.0%（P<0.05）,比长光照组显著高100%（P<0.05）,长光照组与对照组差异不显著（P>0.05）。短光照周期显著提高了下丘脑、垂体和卵巢中褪黑激素受体亚型MT1表达,而不是MT2受体亚型。光照周期调控母兔发情的作用机制可能是褪黑激素通过MT1受体途径实现的。     

Effects of Dietary Supplemental Different Levels of Sugar Beet Pulp on the Digestion and Metabolism of Nutrients of Yanqi Horse

This experiment was conducted to study the effects of supplemental different levels of sugar beet pulp on the nutrients of digestion and metabolism in Yanqi horse.12 Yanqi horses were randomly divided into three groups according to performance,body weight,body measurement and age.The groups were referred to as the control group,group Ⅰ and group Ⅱ,each group with 4 Yanqi horses.Yanqi horses in the control group,group Ⅰ and group Ⅱ were supplied with 0,0.6,1.2 kg/d of sugar beet pulp on the condition of feeding the same basal diet and roughage (oat straw).The total duration of the experiment was 20 days,the first 15 days of the experiment was the preliminary period,and the formal experiment lasted for 5 days.The results showed that compared with the control group,the apparent digestibility of OM,NDF,ADF and digestibility amount of NDF,ADF in group Ⅱ was extremely significantly improved (P<0.01),the digestibility amount of crude protein,calcium and the apparent digestibility of OM in group Ⅱ was significant difference (P<0.05);The digestibility amount of NDF,ADF of group Ⅰ was increased by 8.19% (P<0.05) and 9.43% (P<0.05),respectively,and the apparent digestibility (amount) of OM,crude protein,calcium were no significant difference (P>0.05).Compared with group Ⅰ,the apparent digestibility (amount) of OM,NDF,ADF of group Ⅱ were significantly or extremely significant difference (P<0.05;P<0.01),but the apparent digestibility (amount) of crude protein,calcium were no significant difference (P>0.05).Compared with the control group and group Ⅰ,the digestible engergy of group Ⅱ was increased by 14.92% (P<0.01) and 11.23% (P<0.01),but no significant difference was observed between group Ⅰ and control group (P>0.05);The metabolic engergy of group Ⅱ was increased by 11.73% (P<0.05) and 14.83% (P<0.05),respectively.Therefore,the digestion amount of OM,CP of Yanqi horse and the digestion amount and the apparent digestibility of NDF,ADF could be significantly or extremely significantly improved by the supplementation of 1.2 kg/d of sugar beet pulp,and the ability to use the nutrients were obviously improved for Yanqi horse.     

日粮添加不同水平甜菜粕对焉耆马营养物质消化代谢的影响

试验旨在研究日粮中添加不同水平甜菜粕对焉耆马营养物质消化代谢的影响。选取运动成绩、体重、年龄和体尺相近的焉耆马12匹,随机分为3组,每组4匹。对照组、试验Ⅰ组和试验Ⅱ组在饲喂相同的基础日粮和粗饲料(燕麦秸)的条件下分别补饲0、0.6和1.2 kg/d甜菜粕。试验期20 d,其中预试期15 d,正试期5 d。结果表明,与对照组相比,试验Ⅱ组焉耆马对有机物、NDF、ADF的消化量和对日粮NDF、ADF的表观消化率均有极显著提高(P<0.01),对粗蛋白质、钙的消化量和对有机物的表观消化率影响差异显著(P<0.05);试验Ⅰ组焉耆马对NDF和ADF的消化量分别增加了8.19%、9.43%,均达到了显著水平(P<0.05),而对有机物、粗蛋白质、钙的消化量和表观消化率均差异不显著(P>0.05);试验Ⅱ组焉耆马对有机物、NDF和ADF的消化量和表观消化率与试验Ⅰ组相比,均达到了显著或极显著水平(P<0.05;P<0.01),但对粗蛋白质和钙的消化量和表观消化率影响差异不显著(P>0.05)。试验Ⅱ组焉耆马的消化能与对照组和试验Ⅰ组相比分别提高14.92%(P<0.01)和11.23%(P<0.01),而试验Ⅰ组与对照组相比差异不显著(P>0.05);对代谢能而言,试验Ⅱ组比对照组和试验Ⅰ组分别提高11.73%(P>0.05)和14.83%(P<0.05)。因此,日粮中添加1.2 kg/d的甜菜粕,可显著提高焉耆马对日粮有机物和粗蛋白质的消化量,极显著提高NDF和ADF的表观消化量(率)及消化能,可明显改善焉耆马对营养物质的利用率。     

饲喂水平对冬毛生长期雄性北极狐生产性能、器官发育及机体能量沉积的影响

旨在研究不同饲喂水平对冬毛生长期雄性北极狐生产性能、器官发育及机体能量沉积的影响。本试验选取46只161日龄,平均体重为（7 285±5.77）g健康的雄性北极狐,其中6只北极狐作为试验初屠宰试验对照,另外40只北极狐随机分成4组（每组10个重复,每个重复1只）,分别按照自由采食组（AL,I组）、自由采食量的80%组（IR80,II组）、自由采食量的60%组（IR60,III组）和自由采食量的40%组（IR40,IV组）4个水平饲喂,预饲期7 d,试验期67 d,通过饲养试验、屠宰试验并结合化学分析方法来评定生长性能、毛皮品质、器官发育及机体能量沉积的各项指标。结果表明：1）随饲喂水平降低,平均干物质采食量（ADFI）呈极显著降低（P<0.01）,II组平均日增重（ADG）极显著高于IV组（P<0.01）,显著高于III组（P<0.05）,与I组差异不显著（P>0.05）;饲喂水平对北极狐末重和料重比（F/G）无显著影响（P>0.05）,但II组末重略高于其他组,F/G略低于其他组。2）饲喂水平极显著影响了北极狐鲜皮长（P<0.01）,I组极显著高于III和IV组,与II组无显著差异（P>0.05）;随饲喂水平降低,体长、干皮长、针毛长和绒毛长均呈降低趋势,但各组间均无显著性差异（P>0.05）。3）饲喂水平显著影响了北极狐心脏重和心脏指数（P<0.05或P<0.01）,II组心脏重显著高于III组（P<0.05）,与I组和IV组差异不显著（P>0.05）,IV组心脏指数极显著高于I组（P<0.01）,显著高于III组（P<0.05）,与II组差异不显著（P>0.05）,而饲喂水平对肝、肾、肺和脾重及其脏器指数均无显著影响（P>0.05）。4）随饲喂水平降低,毛皮增重、毛皮脂肪沉积及其能量沉积、毛皮沉积总能量、胴体脂肪沉积及其能量沉积、胴体蛋白沉积及其能量沉积和胴体沉积总能量均呈降低趋势,但饲喂水平对毛皮、胴体能量沉积各项指标均无显著性影响（P>0.05）。采用适当限饲（IR80）不影响冬毛生长期北极狐机体器官的正常发育,能够保证其体重增长及毛皮品质,提高了饲料转化效率,且还降低了机体能量沉积过度带来的肥胖风险。     

运输对伊犁马肌肉损伤和氧化应激的影响

试验旨在研究运输对伊犁马肌肉损伤和氧化应激的影响。选取体重相近的5匹24月龄的伊犁母马,环境适应7 d后开展运输试验,其运输过程中禁食禁水,运输总路程约为400 km,运输时间为8 h。分别于运输0（T1）、4（T2）、8（T3）和卸车后12 h （T4）采集马颈静脉血,分离血清,进行相关生理生化指标的测定。结果显示,与T1相比,肌肉损伤生化标志物肌红蛋白（MYO）和肌钙蛋白（cTnT）在T2时刻显著上升（P<0.05）,T3和T4时刻极显著上升（P<0.01）;乳酸脱氢酶（LDH）在T3时刻显著上升（P<0.05）;炎症反应生化标志物C反应蛋白（CRP）和白细胞介素-6（IL-6）在T2、T3和T4时刻极显著上升（P<0.01）;皮质醇（COR）在T2和T3时刻极显著上升（P<0.01）,在T4时刻显著上升（P<0.05）;总抗氧化能力（T-AOC）、谷胱甘肽过氧化物酶（GSH-Px）和超氧化物歧化酶（SOD）在T2、T3和T4时刻极显著下降（P<0.01）;过氧化氢酶（CAT）在T3和T4时刻显著下降（P<0.05）;活性氧（ROS）在T2和T4时刻显著上升（P<0.05）,在T3时刻极显著上升（P<0.01）;丙二醛（MDA）在T3和T4时刻极显著上升（P<0.01）。因此,运输8 h导致了伊犁马肌肉损伤和氧化应激反应的发生,其不良影响在卸车后12 h内不能消除。     

热休克蛋白HSP90B1影响牛病毒性腹泻病毒复制的研究

旨在探究热休克蛋白90 β家族成员1(heat shock protein 90 beta family member 1,HSP90B1)对牛病毒性腹泻病毒(bovine viral diarrhea virus, BVDV)复制的影响。本研究通过实时荧光定量聚合酶链式反应(quantitative real time polymerase chain reaction, qRT-PCR)和Western blot检测BVDV感染MDBK细胞后HSP90B1 mRNA和蛋白的表达情况,利用CRISPR/Cas9技术构建HSP90B1 KO细胞,计数检测敲除HSP90B1对细胞生长的影响,BVDV TC株感染HSP90B1 KO和对照组Scramble细胞后,使用qRT-PCR、免疫荧光、病毒滴度以及细胞病变效应(cytopathic effects, CPE)检测BVDV的复制情况。结果表明,qRT-PCR检测显示BVDV感染24 h时与空白组相比HSP90B1 mRNA转录水平显著升高(P<0.05),36 h后极显著升高(P<0.01),同时Western blot显...     

不同酶制剂对水稻秸秆和白酒糟混合青贮品质的影响

为研究不同酶制剂对水稻（Oryza sativa L.）秸秆和白酒糟（m︰m=1︰1）混合青贮品质的影响，设置CK（对照）、CEL（250 U·g^-1纤维素酶）和XYL（250 U·g^-1木聚糖酶）3个处理，青贮120 d后取样测定其青贮品质。结果表明：CEL处理的感官评定等级为良好；CEL和XYL处理的干物质、粗脂肪、钾和镁含量均显著高于CK（P<0.05），且CEL处理的粗蛋白和磷含量显著高于CK（P<0.05）；CEL和XYL处理的中性洗涤纤维、酸性洗涤纤维、木质素和纤维素含量均显著低于CK（P<0.05），且CEL处理显著低于XYL处理（P<0.05）；CEL和XYL处理30 h的养分消化率显著高于CK（P<0.05），且CEL处理48 h的养分消化率均显著高于CK（P<0.05）；CEL和XYL处理的饲用价值显著高于CK（P<0.05），且CEL处理显著高于XYL处理（P<0.05）。综上所述，添加纤维素酶、木聚糖酶能够提高水稻秸秆和白酒糟混合青贮的营养品质、养分消化率和饲用价值，且纤维素酶的添加效果优于木聚糖酶。     

Effects of KDM1A on the Meiotic Maturation and Developmental Potential of Yak Oocytes

The purpose of this study was to explore the effects of KDM1A on the meiotic maturation and developmental potential of yak oocytes. The specific inhibitor GSK-KDM1A of KDM1A was added into in vitro maturation medium of yak oocytes. After 24 h in vitro culture of yak cumulus oocyte complexes (COCs), the expansion of cumulus cells and the extrusion of the first polar body were observed. The expression level of KDM1A in oocyte was measured by immunofluorescence during in vitro culture. The expression levels of Kdm1a, Oct-4, Sox-2 and Nanog in oocytes were detected by RT-qPCR. Then, yak oocytes were fertilized after in vitro culture, and the cleavage rate and blastocyst formation rate were observed, respectively. The results showed that the cumulus cells expansion in GSK-KDM1A groups were significant lower than those in control group (P<0.05) after 24 h culture, and the cumulus cells expansion and the first polar body extrusion rate in 320 nmol·L^-1 group were significantly lower than those in 160 nmol·L^-1 group (P<0.05). During oocyte in vitro maturation, Kdm1a showed dynamic expression profile, and the expression level of Kdm1a in MⅠ-stage was significantly lower than that in GV-stage and MⅡ-stage (P<0.05). The GSK-KDM1A could significantly inhibit the expression of KDM1A protein in oocytes (P<0.05), and the expression of KDM1A in 320 nmol·L^-1 group was significantly lower than that in 160 nmol·L^-1 group (P<0.05). The expression levels of Oct-4 and Sox-2 in GSK-KDM1A group were significantly higher than that in control group (P<0.05), but there was no significant difference in the expression of Nanog (P>0.05). After 24 h culture, the cleavage rates of oocytes in GSK-KDM1A groups were significantly lower than those in control group (P<0.05), while the blastocyst formation rate was not significantly different (P<0.05). In conclusion, KDM1A is involved in regulating the meiotic maturation process of yak oocytes. GSK-KDM1A can effectively inhibit the expression of KDM1A, affect the meiotic maturation and developmental potential of oocytes, which reveal that KDM1A plays an important role in this process.     

玉米赤霉烯酮对后备母猪子宫和卵巢抗氧化和炎症指标及相关基因表达的影响

试验旨在研究玉米赤霉烯酮（zearalenone,ZEA）对后备母猪子宫和卵巢抗氧化和炎症指标及相关基因表达的影响。选择胎次和体重（（23.20±0.68）kg）相近的长×大二元后备母猪48头,随机分为4组,每组设12个重复,每个重复1头母猪。对照组（CON组）饲喂基础饲粮,试验组（T1、T2、T3组）饲喂在基础饲粮中分别添加200、800、1600 μg·kg^-1ZEA的试验饲粮。预试期7 d,正试期40 d。结果如下：1）与CON组相比,T3组血清T-AOC和SOD活性显著降低（P<0.05）,MDA水平显著升高（P<0.05）。2）与CON组相比,子宫组织中,T2和T3组T-AOC活性显著降低（P<0.05）,T3组SOD活性极显著降低（P<0.01）,卵巢组织中,T3组T-AOC活性、T2组GSH-Px和SOD活性显著降低（P<0.05）,T3组MDA水平显著升高（P<0.05）。3）与CON组相比,T2组血清TNF-α和IL-4水平显著升高（P<0.05）,T1~T3组血清IL-10水平显著降低（P<0.05）。4）与CON组相比,子宫组织中,T2组IL-1β水平显著升高（P<0.05）,T1和T3组IL-10水平显著降低（P<0.05）。卵巢组织中,T2组TNF-α水平显著升高（P<0.05）,T1~T3组IL-4水平显著升高（P<0.05）。5）子宫组织中,T3组Cu/Zn-SOD mRNA的相对表达显著低于CON组（P<0.05）,T2组IL-1β mRNA的相对表达显著高于CON组（P<0.05）。6）卵巢组织中,T3组GSH-Px mRNA的相对表达显著低于CON组（P<0.05）,T2组IL-1β mRNA的相对表达显著高于CON组（P<0.05）。综上所述,ZEA能通过抑制SOD和GSH-Px的表达与合成,降低后备母猪子宫和卵巢的抗氧化性能,并引起氧化应激。ZEA能通过促进TNF-α和IL-1β表达与合成引起子宫和卵巢的炎症反应,抑制IL-10的表达与合成从而抑制两组织的抗炎反应。     

Study on the Anti-Escherichia coli Activity and Mechanism of Water Extract of Radix isatidis Powder

In order to explore the antibacterial activity and mechanism of the Radix isatidis powder water extract,the effects of the Radix isatidis powder water extract on the morphology and structure of Escherichia coli (E.coli) were tested by scanning electron microscopy and transmission electron microscopy.The effects of the Radix isatidis powder water extract on the conductivity and total leakage rate of E.coli and the content of alkaline phosphatase in the culture medium of the content of protein,DNA and RNA were stained by DAPI to detect the effect of the Radix isatidis powder water extract on the nucleic acid of E.coli,and the effect of the Radix isatidis powder water extract on the metabolism of E.coli in vivo,in vitro,ALT,AST,pyruvic acid and ATP.The results showed that after the Radix isatidis powder water extract acted on E.coli for 10 h,it was observed by SEM that the bacteria appeared to overflow and shrink,the length of the bacteria became shorter obviously,many residues were formed due to breakage,some of them were sunken in the middle and deformed.Under TEM,it was observed that the boundary of the cell wall of E.coli was unclear,the wall membrane was zigzag,deformed and some of the bacteria were broken.The protein content outside the cell was significantly different from that in the blank control group from 8 h (P<0.01), and that in the cell from 4 h (P<0.01). DNA content had no significant difference with blank control group before 12 h (P>0.05), but had significant difference with blank control group from 16 h (P<0.01); RNA content began to decrease at 8 h, and was significantly different from blank control group at 12 h (P<0.05), and was extremely significant from 16 h (P<0.01). There was no significant difference between ALT and AST (P>0.05). The pyruvate content in culture medium and bacteria was higher than that in blank control group, and the difference was very significant from 4 h (P<0.01). The ATP content in the culture medium was significantly different from that in the blank control group (P<0.01), and the ATP content in the cell was significantly different from that in the blank control group from 4 h (P<0.01).In conclusion,the Radix isatidis powder water extract could inhibit the synthesis and metabolism of bacterial genetic material and the content of pyruvate and ATP by destroying the integrity of cell wall and cell membrane.     

光照节律对肉兔生长性能、血清生化指标、肉品质和动物福利的影响

旨在研究光照节律对肉兔生长性能、肉品质、血清生化指标和行为的影响。选取135只35日龄断奶伊拉公兔,随机分为3个处理,每个处理15个重复,每个重复3只肉兔,设置12 h光暗循环(12L∶12D)、24 h光照(24L)和24 h黑暗(24D)的3种光照节律,光照强度为100 lx,预试期为7 d,试验期为35 d。结果表明,与12L∶12D组相比,24D组肉兔21~35 d的平均日采食量(ADFI)、饲料转化率(FCR)、半净膛率、背最长肌pH和熟肉率显著提高(P<0.05),血糖浓度显著降低(P<0.05),其中FCR和半净膛率在各个处理组中最高(P<0.01)。24L组的皮指数显著低于12L∶12D和24D组(P<0.01),而白蛋白(ALB)和肌肉失水率显著高于于12L∶12D和24D组(P<0.01)。各个处理组间肌肉肉色以及肌肉常规营养指标没有显著差异(P>0.05)。24D组的肉兔坐的行为持续时间最少(P<0.01),啃咬行为频率较低(P<0.05),运动、直立、站立行为频率最高(P<0.05)。结果显示,长时间黑暗环境可以提高肉兔的生长性能,降低血糖浓度。同时,肉兔在长期的黑暗中表现出更多的社会性、较少的恐惧和焦虑,显示出良好的动物福利。     

Effects of Carvacrol and Thymol on Colonic Microorganisms in Weaned Piglets

This study was conducted to investigate the effects of carvacrol and thymol on the relative abundance of colonic microorganisms in weaned piglets and their metabolic pathways.A total of sixty four 28-day-old (8.5 kg±1.0 kg) weaned piglets were divided into 4 groups with 16 replicate of 1 piglet:Group A (control group,basal diet),group B (basal diet＋20 g/T carvacrol),group C (basal diet＋20 g/T thymol),and group D (basal diet＋20 g/T carvacrol-thymol blend (mass ratio 2:1)).The experiment lasted 30 d.The sequencing was taken based on the Illumina HiSeq sequencing platform,and using paired-end sequencing (Paired-End) method.Perform species annotation and abundance were analysized by clustering OTUs,and alpha diversity analysis、beta diversity analysis、significant species difference analysis and functional gene prediction analysis were further conducted.The results showed as follow:①The colonic microorganisms biodiversity of the experimental groups were significantly higher than that of control group (P<0.05).②The relative abundance of Firmicutes in the colon of group D was extremely significantly higher than that of control group (P<0.01),significantly higher than groups B and C (P<0.05),the relative abundance of Bacteroidetes was significantly higher than that of control group and group C (P<0.05),and the relative abundance of Epsilonbacteraeota,Spirochaetes and Proteobacteria were extremely significantly lower than that of control group (P<0.01).The relative abundance of Campylobacter in the colon of group D was extremely significantly lower than that of control group (P<0.01),that of Rikenellaceae_RC9_gut_group in group D was significantly higher than that of group B (P<0.05),that of Roseburia in group D was extremely significantly higher than that of group A (P<0.01),that of Helicobacter was extremely significantly lower than that of group A (P<0.01).③The lipid metabolism level of colonic microorganisms in group D was extremely significantly higher than that of control group (P<0.01),and colonic microbial carbohydrate metabolites,biosynthetic of other secondary metabolites and other amino acid metabolism level were significantly higher than that of control group (P<0.05),and energy metabolism level of colonic microorganisms was extremely significantly lower than that of control group (P<0.01).In conclusion,under the condition of this experiment,dietary supplementation with 20 g/t carvacrol-thymol improved the microbial community structure of colonic microorganisms in weaned piglets.The relative abundance of colonic beneficial bacteria was increased,the relative abundance of colonic harmful bacteria was decreased,and positively affected the metabolic function of colonic microorganisms.     

板蓝根微粉水提物抗大肠杆菌活性及其机制的探究

为了探究板蓝根微粉水提物的抗菌活性及其抗菌机制，试验通过扫描电镜、透射电镜检测板蓝根微粉水提物对大肠杆菌形态和结构影响；酶标仪测定板蓝根微粉对大肠杆菌电导率、胞内物质总漏出率影响；测定大肠杆菌培养液中碱性磷酸酶含量以及大肠杆菌菌体内、外蛋白质含量；通过DAPI染色DNA、RNA，检测板蓝根微粉水提物对大肠杆菌核酸的影响；检测板蓝根微粉水提物对大肠杆菌菌体内、外谷丙转氨酶（ALT）、谷草转氨酶（AST）、丙酮酸以及三磷酸腺苷（ATP）等菌体内代谢的影响。结果显示，板蓝根微粉水提物作用大肠杆菌10 h后，扫描电镜检测可见菌体出现溢缩，菌体长度明显变短，断裂形成许多残体，有的中间凹陷，发生变形；透射电镜下可观察大肠杆菌的胞壁界限模糊不清，壁膜呈现锯齿状，弯弯曲曲，变形，有的菌体破碎。总漏出率、电导率以及碱性磷酸酶含量测定结果显示，板蓝根微粉水提物各组D_{600 nm}均高于空白对照组，且呈剂量依赖性。菌体外蛋白质含量从8 h开始与空白对照组差异极显著（P<0.01）；菌体内蛋白质含量从4 h开始与空白对照组差异极显著（P<0.01）。DNA含量在12 h前与空白对照组无显著差异（P>0.05），从16 h开始与空白对照组差异极显著（P<0.01）；RNA含量在8 h开始降低，在12 h时与空白对照组差异显著（P<0.05），从16 h开始差异极显著（P<0.01）。ALT和AST浓度测定无显著性差异（P>0.05）。培养液和菌体内的丙酮酸含量均高于空白对照组，且从4 h开始与空白对照组差异极显著（P<0.01）。培养液中的ATP含量与空白对照组差异极显著（P<0.01）；菌体内ATP含量从4 h开始与空白对照组差异极显著（P<0.01）。综上，板蓝根微粉水提物可以通过破坏细胞壁、细胞膜的完整性，抑制细菌遗传物质合成和代谢，影响丙酮酸和ATP含量从而实现抗大肠杆菌作用。