HMGB1与日本血吸虫病肝炎症和纤维化的相关性分析

旨在探究高迁移率族蛋白B1（high mobility group box 1,HMGB1）与日本血吸虫病肝炎症和纤维化的相关性,本研究将BALB/c小鼠随机分为空白对照组（n＝10）,感染后4周组、6周组和HMGB1抑制剂组（n＝5）,进行血常规及生化检测,观察肝病理学及Masson染色情况、荧光定量PCR检测HMGB1、IL-1β、TNF-α、IL-6、IFN-γ、α-SMA、Col1α1、TGF-β1、Smad2和Smad3的转录水平。结果显示：感染后4周,小鼠谷丙转氨酶（alanine aminotransferase,ALT）、肝组织HMGB1、α-SMA、TGF-β1、IL-6和 TNF-α的转录水平均显著或极显著升高（P<0.05或P < 0.01）;感染后6周,小鼠WBC、Neu、Mon、Eos、谷草转氨酶（aspartate aminotransferase,AST）和ALT极显著升高（P<0.01）,肝组织HMGB1、IL-1β、TNF-α、IL-6、IFN-γ、α-SMA、Col1α1、TGF-β1、Smad2和Smad3的转录水平均显著或极显著升高（P<0.05或P<0.01）;感染后6周小鼠肝虫卵聚积处伴有炎性细胞浸润和胶原纤维增生;HMGB1抑制剂可显著降低小鼠血清ALT值和肝组织HMGB1、α-SMA、TGF-β1、IL-6和TNF-α的转录水平（P<0.05）。综上表明,HMGB1与日本血吸虫病肝炎症和纤维化具有一定相关性。     

植物精油和柠檬酸复合物对肉鸡肠道屏障功能及炎症反应的影响

本试验通过研究饲粮中添加不同水平植物精油和柠檬酸复合物对肉鸡肠道屏障功能、血清细胞因子含量和关键免疫蛋白基因表达的影响,旨在探索植物精油和柠檬酸复合物在肉鸡生产上的适宜添加剂量。选用1日龄健康科宝肉鸡1 344只,随机分为4组（每组12个重复,每个重复28只鸡）,对照组饲喂基础饲粮,3个试验组（T1、T2、T3）分别饲喂添加0.8、1.0和1.5 kg/t的包被植物精油和柠檬酸复合物试验饲粮,试验期为42 d。试验结束当天,每个重复选取6只接近平均体重的肉鸡翅静脉采血,然后屠宰并取空肠、脾脏、肠系膜淋巴结（MLNs）。分别测定血清中D-乳酸（D-LA）、细胞因子的含量及二胺氧化酶（DAO）活性;并对空肠黏膜紧密连接蛋白以及脾脏和MLNs中Toll样受体（TLRs）的mRNA表达量进行测定。结果表明：与对照组相比,T2组肉鸡血清中D-LA含量和DAO活性均极显著降低（P<0.01）;T3组肉鸡血清中D-LA含量和DAO活性均显著降低（P<0.05）。T2和T3组肉鸡空肠封闭蛋白1（Claudin-1）的mRNA相对表达量均显著升高（P<0.05）,T2组闭合小环蛋白1（ZO-1）的mRNA相对表达量显著升高（P<0.05）。T1组血清中IL-1β含量显著降低（P<0.05）,MLNs中TLR4 mRNA表达量显著降低（P<0.05）;T2组血清中IL-1β含量极显著降低（P<0.01）,IL-2含量显著提高（P<0.05）,脾脏和MLNs中TLR4 mRNA相对表达量极显著或显著降低（P<0.01;P<0.05）;T3组血清中IL-1β含量极显著降低（P<0.01）,TNF-α含量显著降低（P<0.05）,IL-2含量显著升高（P<0.05）,脾脏和MLNs中TLR4 mRNA相对表达量极显著降低（P<0.01）。与T1组相比,T2组血清中D-LA含量和DAO活性均显著降低（P<0.05）,TNF-α含量显著降低（P<0.05）;T3组血清中IL-1β含量显著降低（P<0.05）。综上所述,饲粮中添加植物精油和柠檬酸复合物能够提高肉鸡肠道紧密连接蛋白的表达,降低肠道通透性,下调促炎细胞因子的表达,进而增强肠道屏障功能,缓解机体炎症。根据本试验结果,推荐科宝肉鸡饲粮植物精油和柠檬酸复合物适宜添加量为1.0 kg/t。     

复方中药发酵液对肉鸡大肠杆菌病的防治效果及免疫调节作用

试验旨在研究复方中药发酵液对肉鸡大肠杆菌病的防治效果及免疫调节作用。选择健康1日龄AA白羽肉鸡240羽,分别进行大肠杆菌O78半数致死剂量和复方中药发酵液对大肠杆菌病的防治试验。在28日龄评价治疗效果,并在28、42日龄分别检测免疫器官指数、抗氧化指标和肠道菌群等指标。结果显示,经过植物乳杆菌和枯草芽孢杆菌等菌株发酵后的复方中药发酵液能够将大肠杆菌的感染时间由4 h延迟至24 h,高剂量组（1.0 g/mL）、中剂量组（0.5 g/mL）、抗生素组（硫酸新霉素）有效率分别为80.0%、76.7%和80.0%,治愈率分别为80.0%、70.0%和70.0%;高剂量组与感染组和抗生素组相比,28日龄时胸腺指数分别提高了165.6%（P<0.05）、25.4%（P>0.05）,法氏囊指数分别提高了82.6%和94.3%（P<0.05）,42日龄时胸腺指数分别提高了97.4%和82.0%（P<0.05）,法氏囊指数分别提高了43.4%和27.5%（P<0.05）,28、42日龄时抗氧化指标T-AOC、TP含量及GSH-Px、T-SOD活性差异显著（P<0.05）,能够显著提高肠道内乳酸菌含量,降低大肠杆菌含量（P<0.05）。综上,中剂量组（0.5 g/mL）复方中药发酵液抗菌效果明显,能够替代抗生素,高剂量组（1.0 g/mL）复方中药发酵液对肉鸡其他指标明显优于抗生素。     

葛根素对饲喂氧化大豆油肉鸡小肠黏膜屏障功能及抗氧化能力的影响

本试验旨在研究葛根素对饲喂氧化大豆油饲粮黄羽肉鸡小肠黏膜形态结构、紧密连接蛋白基因表达及抗氧化能力的影响。试验采用2×2因子设计,因子包括油脂质量（新鲜大豆油和氧化大豆油）和葛根素添加水平（0、750 mg·kg^-1）。选取健康1日龄雌性黄羽肉鸡360只,随机分成4个处理组,分别为新鲜大豆油饲粮组、新鲜大豆油+葛根素（750 mg·kg^-1）饲粮组、氧化大豆油饲粮组和氧化大豆油+葛根素（750 mg·kg^-1）饲粮组,每组6个重复,每个重复15羽。在28和56日龄时,每个重复随机选取1只鸡,取十二指肠、空肠、回肠检测小肠形态结构和测定小肠黏膜紧密连接蛋白基因表达及抗氧化指标。结果表明：1）饲喂氧化大豆油显著降低28日龄肉鸡十二指肠的绒毛高度和绒毛高度/隐窝深度（V/C）、回肠V/C和claudin-1 mRNA表达量以及56日龄肉鸡3个肠段的绒毛高度及V/C （P<0.05）,显著升高56日龄3个肠段的隐窝深度（P<0.05）。饲粮中添加葛根素显著提高28日龄肉鸡空肠绒毛高度和V/C、回肠闭合小环蛋白1（ZO-1） mRNA表达量和56日龄肉鸡十二指肠V/C、空肠和回肠的绒毛高度及V/C （P<0.05）,显著降低3个肠段的隐窝深度（P<0.05）。2）饲喂氧化大豆油显著升高28日龄十二指肠还原型谷胱甘肽（GSH）含量和空肠还原型谷胱甘肽过氧化物酶（GSH-Px）活性（P<0.05）,并显著降低28日龄回肠GSH含量、超氧化物歧化酶（SOD）活力和总抗氧化能力（T-AOC）（P<0.05）。饲粮中添加葛根素显著提高28日龄肉鸡十二指肠GSH-Px活性（P<0.05）。饲喂氧化大豆油显著降低56日龄回肠SOD活性和T-AOC （P<0.05）,饲粮中添加葛根素显著提高56日龄肉鸡十二指肠SOD和回肠SOD、GSH-Px活性（P<0.05）,显著降低56日龄肉鸡空肠GSH含量、SOD活性、T-AOC和回肠T-AOC （P<0.05）。综上所述,饲喂氧化大豆油破坏肠道黏膜形态结构、降低紧密连接蛋白基因的表达量和抗氧化能力,添加葛根素可提高肉鸡小肠黏膜紧密连接蛋白基因的表达量,改善氧化损伤条件下肠道黏膜形态结构,提高抗氧化酶的活性而提高其抗氧化能力。     

丁酸梭菌对肉鸡钙磷代谢和胫骨指标的影响

试验通过在基础日粮中添加丁酸梭菌,研究其对肉鸡钙磷代谢和胫骨指标的影响。选择1日龄180只健康AA肉鸡,随机分入3个处理组,每个处理组6个重复,分别为对照组、抗生素组（ATB）和丁酸梭菌组（CB）。对照组饲喂基础日粮,ATB组在基础日粮中添加5 mg/kg黄霉素、75 mg/kg金霉素和20 mg/kg吉他霉素,CB组在基础日粮中添加2.5×10⁸ CFU/kg丁酸梭菌,试验期42 d。结果显示,与对照组相比,①CB组肉鸡血清钙（Ca）、磷（P）含量差异不显著（P>0.05）,21日龄ATB组肉鸡血清钙含量显著提高（P<0.05）;②21日龄CB组肉鸡胫骨强度显著提高（P<0.05）;42日龄CB组肉鸡胫骨磷含量极显著提高（P<0.01）;③21日龄CB组肉鸡排泄物中磷含量极显著降低（P<0.01）,并与ATB组无显著差异;42日龄CB组肉鸡排泄物中钙含量极显著提高（P<0.01）,排泄物中磷含量显著提高（P<0.05）。综合上述结果,在AA肉鸡基础日粮中添加2.5×10⁸ CFU/kg丁酸梭菌有利于改善肉鸡的胫骨发育,改善肉鸡对钙、磷的吸收利用,能有效降低21日龄肉鸡钙、磷排泄量,但并未降低42日龄肉鸡排泄物中钙、磷含量。     

蜂胶对细菌脂多糖刺激下奶牛乳腺上皮细胞炎症相关基因mRNA转录水平和紧密连接蛋白的影响

本试验旨在研究中国蜂胶乙醇提取物（ethanol extract of Chinese propolis，EECP）对细菌脂多糖（lipopolysaccharide，LPS）刺激下体外培养奶牛乳腺上皮细胞炎症相关基因mRNA转录水平和紧密连接渗透性的影响。EECP中总酚酸和总黄酮含量测定采用福林酚法和硝酸铝法，并建立LPS诱导奶牛乳腺上皮细胞（bovine mammary epithelial cells，MAC-T）炎症模型，采用CCK-8法测定EECP对MAC-T相对增殖率的影响，利用实时荧光定量PCR（RT-qPCR）评估EECP对LPS诱导的MAC-T细胞炎症相关因子（IL-6、IL-8、TNF-α和IL-1β）相对mRNA转录水平；以及对紧密连接蛋白（occludin、ZO-1）相对mRNA转录水平进行检测，并进一步利用免疫荧光技术对紧密连接膜蛋白进行定位，确定EECP对LPS诱导MAC-T细胞炎症紧密连接渗透性的影响。结果显示：EECP中总酚酸含量为106.35 mg没食子酸当量（GAE）·g^-1、总黄酮含量为320.85 mg芦丁当量（RE）·g^-1；CCK-8结果显示EECP的安全浓度为0~15 μg·mL^-1，并可有效提高LPS刺激下MAC-T的活力；LPS刺激显著增加了细胞炎症相关因子IL-6、IL-8、TNF-α和IL-1β mRNA的转录量（P<0.001）；但2.5~15.0 μg·mL^-1 EECP预处理显著降低了IL-6、IL-8、TNF-α和IL-1β mRNA的转录量；与此类似，LPS刺激显著抑制了紧密连接蛋白基因（occludin、ZO-1）mRNA的转录量（P<0.01），而EECP预处理后紧密连接蛋白基因（occludin和ZO-1）mRNA的转录量显著增加（P<0.05）；免疫荧光染色试验也证实EECP能通过上调紧密连接蛋白（occludin、ZO-1）的表达，缓解LPS诱导的乳腺上皮细胞屏障功能紊乱。该结果证实，EECP对细菌脂多糖诱导奶牛乳腺上皮细胞炎症具有良好的保护作用，这为利用中国蜂胶预防奶牛乳腺炎提供了试验基础。     

不同月龄红河黄牛血液生化和免疫指标分析

为明确放养条件下红河黄牛的营养和免疫状况，选取60头红河黄牛，公母各半，按性别和年龄（12、36和60月龄）分成6个组，颈静脉采血分离血清，测定生化和免疫指标。结果显示：①相同性别的黄牛血清总蛋白（TP）和球蛋白（GLOB）含量12月龄显著低于60月龄（P<0.05），母牛碱性磷酸酶（ALP）活性和血糖（GLU）含量12月龄显著高于60月龄（P<0.05），其他血清生化指标在不同月龄间均无显著差异（P>0.05）。公、母牛血清TP和GLOB含量均随月龄增加而升高，而ALP活性随月龄增加而降低。不同月龄母牛血清TP、白蛋白（ALB）、GLOB含量和谷丙转氨酶（ALT）活性均高于公牛（P<0.05）；而ALP活性和GLU含量公牛高于母牛（P<0.05）。②除母牛血清IL-8在12和36月龄间存在显著差异（P<0.05）外，相同性别的黄牛血清白细胞介素（IL）-2、IL-8、IL-10、免疫球蛋白（Ig） A、IgG、IgM、γ干扰素（IFN-γ）、肿瘤坏死因子α（TNF-α）含量在3个月龄间均无显著差异（P>0.05）。不同月龄母牛IL-2、IgA、IgG、IgM、IFN-γ和TNF-α含量均高于公牛（P<0.05）。综合以上结果，红河黄牛公、母牛随月龄增加，血清TP和GLOB含量显著增加，而ALP活性降低；母牛血清生化和免疫指标总体高于公牛。     

发酵棉粕对肉仔鸡生长性能、屠宰性能、营养物质表观消化率和脂肪沉积的影响

试验旨在研究棉粕经热带假丝酵母菌发酵后得到的发酵棉粕对白羽肉鸡生长性能、屠宰性能、营养物质表观消化率和脂肪沉积的影响。试验选取180羽1日龄科宝白羽肉鸡,随机分成3组,每组6个重复,每个重复10羽鸡。对照组饲喂不加菌发酵棉粕（CSM）,两个试验组分别饲喂热带假丝酵母菌发酵棉粕（FCSM）、CSM+热带假丝酵母菌（CT-USCM,活菌量同FCSM）。21日龄试验结束。结果表明：①FCSM组肉鸡末重、平均日增重均显著高于对照组（P<0.05）,FCSM和CT-USCM组肉鸡的料重比显著低于对照组（P<0.05）;②CT-USCM组肝重率显著高于对照组（P<0.05）,FCSM和CT-USCM组的腹脂率和皮下脂肪厚度显著低于对照组（P<0.05）;③FCSM和CT-USCM组肉鸡干物质和粗蛋白质表观消化率均显著高于对照组（P<0.05）,粗脂肪和粗灰分表观消化率在各组间差异不显著（P>0.05）;④FCSM和CT-USCM组肉鸡血糖（Glu）、甘油三酯（TG）和低密度脂蛋白（LDL）浓度均显著低于对照组（P<0.05）;⑤FCSM和CT-USCM组肉鸡肝脏和腹脂中激素敏感酯酶（HSL）活性和生长激素（GH）水平均显著高于对照组（P<0.05）,两组之间差异不显著（P>0.05）;FCSM组肝脏和腹脂中乙酰辅酶A羧化酶（ACC）和苹果酸酶（ME）活性均显著低于对照组（P<0.05）,而肝脏中脂蛋白酯酶（LPL）活性均显著高于对照组（P<0.05）,腹脂中脂肪酸合成酶（ME）活性显著低于其他两组（P<0.05）。综上所述,在日粮中添加FCSM和热带假丝酵母菌均能不同程度地促进肉鸡生长性能、屠宰性能,提高营养物质表观消化率,同时还能通过影响脂肪代谢相关酶活和激素参与肉鸡脂肪合成和分解,降低肉鸡腹脂率和皮下脂肪厚度。     

中药复方对大肠杆菌感染雏鸭的防治效果研究

本研究对由地锦草、石榴皮等组成的中药复方对大肠杆菌感染雏鸭的临床治疗效果进行了研究，旨在为临床大肠杆菌感染提供新的治疗手段。试验选取50只健康1日龄雏鸭，随机分为5组，每组10只。试验组分别为空白对照组、模型组、阿莫西林组、中药复方高剂量组（DS_H）和低剂量组（DS_L）。给药组在3日龄开始给药，连续给药至试验结束，共10 d。除空白对照组外，在雏鸭6日龄时用大肠杆菌腹腔注射进行攻毒，0.5 mL/只。比较各试验组雏鸭的病死率；对病死鸭和各组存活鸭进行解剖，观察肝脏和小肠的病理剖检及HE染色切片；检测血常规及血清生化各项指标，评价中药复方对感染大肠杆菌雏鸭的治疗效果。结果显示，DS_H、DS_L和阿莫西林均能够有效降低感染雏鸭的死亡率；与模型组相比，阿莫西林和DS_H、DS_L组均可减少肝脏、小肠等器官的损伤。与空白对照组相比，模型组的谷草转氨酶（AST）和谷丙转氨酶（ALT）显著升高（P<0.05），碱性磷酸酶（ALP）、乳酸脱氢酶（LDH）和尿素（UREA）极显著升高（P<0.01），且模型组和给药组血液中白细胞（WBC）、红细胞（RBC）、血红蛋白（HGB）、血小板（PLT）等指标变化显著（P<0.05）。与模型组相比，给药组ALP显著或极显著降低（P<0.05；P<0.01），血液中WBC、RBC、HGB、PLT等指标极显著升高（P<0.01），说明给药组机体损伤程度低于模型组。以上结果表明，中药复方可以减轻因大肠杆菌所引起的机体损伤，且作用效果与阿莫西林相近。     

凝结芽孢杆菌对肉鸡生长性能、血清生化指标和胫骨强度指标的影响

本试验旨在探究凝结芽孢杆菌对肉鸡生长性能、血清生化指标及胫骨强度指标的影响。选取180只1日龄健康AA肉仔公鸡,随机分为3个处理,分别为对照组、抗生素对照组及凝结芽孢杆菌处理组。对照组饲喂基础饲粮,抗生素组在基础饲粮中添加5 mg·kg^-1黄霉素、20 mg·kg^-1吉他霉素和75 mg·kg^-1金霉素,凝结芽孢杆菌处理组在基础饲粮中添加3.6×10⁹ CFU·kg^-1的凝结芽孢杆菌,试验期为42 d。结果表明：1）饲粮中添加凝结芽孢杆菌显著降低了肉鸡前期（1~21日龄）平均日采食量（P<0.05）,对平均体重和平均日增重无显著影响（P>0.05）;2）凝结芽孢杆菌处理组和抗生素组显著增加了肉鸡前期（1~21日龄）血清中总胆固醇和甘油三脂的含量（P<0.05）,并显著提高了血清碱性磷酸酶的活性（P<0.05）;3）相比于对照组和抗生素组,凝结芽孢杆菌显著增加了肉鸡前期（1~21日龄）血清中总蛋白、白蛋白及球蛋白的含量（P<0.05）,并显著降低了血清中尿酸和血氨的含量（P<0.05）,对42日龄血清总蛋白和白蛋白含量无显著影响,但降低了血氨含量;4）饲粮中添加凝结芽孢杆菌显著提高了肉鸡前期胫骨钙含量（P<0.05）,对胫骨灰分和磷含量无显著影响;凝结芽孢杆菌显著提高了42日龄肉鸡胫骨钙和磷的含量（P<0.05）,其胫骨强度在三个处理组中最高,但差异未达到显著水平（P>0.05）。综上所述,肉鸡饲粮中添加凝结芽孢杆菌可在不影响肉鸡前期日增重的前提下降低平均日采食量,提高饲料转化效率,促进脂类和蛋白质在肉鸡体内的代谢与周转,并促进钙、磷在胫骨中的沉积。     

骨碎补总黄酮对缺氧环境中犬骨髓间充质干细胞成骨分化潜能的影响

拟研究骨碎补总黄酮对缺氧环境中犬骨髓间充质干细胞(BMSCs)成骨分化潜能的影响.运用骨碎补总黄酮(TFDR)干预低氧浓度(10％)环境中犬BMSCs 4周后诱导成骨分化,倒置显微镜观察茜素红染色BMSCs钙结节形成,比色法检测碱性磷酸酶(ALP)活性水平,流式细胞术检测细胞线粒体膜电位,激光共聚焦显微镜和RT-PCR...     

Construction of SRD5A2 Gene Interference Vector and Its Effect on the Expression of Lambing Related Genes in Qianbei Ma Goats

The purpose of this study was to investigate the effect of SRD5A2 gene on the expression of genes related to goat lambing traits. In this study, 3 healthy (2-3 years old) and female Qianbei Ma goat with a weight of approximate 32 kg were selected. Ovary tissue was collected and granulosa cells were successfully cultured. Four pairs of siRNA interference sequence of SRD5A2 gene sequence and a pair of negative control sequence were designed by online software. After connecting pGPH1/GFP/Neo vector, the constructed interference vectors were transfected into ovarian granulosa cells of Qianbei Ma goats. The interference vector with optimal interference efficiency was screened by qRT-PCR, and its effect on the expression of SRD5A2 gene was detected. The effects of silencing SRD5A2 gene on the expression of bone morphogenetic protein 15 (BMP15), growth differentiation factor 9 (GDF9), bone morphogenetic protein-1B (BMPR-1B) and follicle-stimulating hormone lactin (FSHβ) were investigated. In this experiment, the interference vector of SRD5A2 gene was successfully constructed on the basis of cultivating ovarian granulosa cells of Qianbei Ma goats. shRNA-SRD5A2-1 interference vector with the optimal interference efficiency was screened out (P<0.01). After inhibiting SRD5A2 gene expression, the expression levels of BMP15, BMPR-1B, GDF9 and FSHβ were significantly reduced. The expression level of BMP15 was extremely significantly lower than that in shRNA-NC control group (P<0.01), and the expression levels of BMPR-1B, GDF9 and FSHβ were significantly lower than those in shRNA-NC control group (P<0.05). In this study, the constructed SRD5A2 gene interference vector was successfully transfected into ovarian granulosa cells. The silencing of SRD5A2 gene can inhibit the expression of genes related to lambing traits, which indicate that SRD5A2 gene is closely related to lambing traits of Qianbei Ma goats, the result will provide a basis for further studies on the regulatory mechanism of SRD5A2 gene on lambing traits in Qianbei Ma goats.     

Study on the Prevention and Control Effect of Chinese Herbal Compound on E.coli Infected Ducklings

In this study,the clinical therapeutic effect of Chinese herbal compound composed of Euphorbia humifusa Willd.and Punica granatum L.on ducklings infected with Escherichia coli (E.coli) was studied in order to provide a new therapeutic method for clinical treatment of E.coli.Fifty healthy one-day-old ducklings were selected and randomly divided into five groups,ten ducklings in each group.The experimental groups were blank control group,model group,amoxicillin group,Chinese medicine compound high-dose group(DS_H) and low-dose group(DS_L).The administration group started administration at 3 days of age,continued administration for 3 days,and continued administration until the end of the experiment,10 days total.Except for control group,when the ducklings were 6 days old,E. coli was intraperitoneally injected for challenge,0.5 mL/head.The mortality of the experimental groups was compared.The dead ducks and the surviving ducks were dissected,the pathological section of liver and small intestine and HE staining sections were observed.The blood routine indexes and serum biochemical analysis were detected to evaluate the therapeutic effect of traditional Chinese medicine compound on E.coli infected ducklings.The results showed that the high and low doses of the traditional Chinese medicine compound and the amoxicillin could effectively reduce the mortality of infected ducklings.Compared with model group,amoxicillin and traditional Chinese medicine high and low dose groups could reduce liver,small intestine and other organs damage.Compared with control group,aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in model group increased significantly (P<0.05),and alkaline phosphatase (ALP),lactate dehydrogenase (LDH),UREA also showed extremely significant upward trend (P<0.01),and the WBC,RBC,HGB,PLT and other indicators changed extremely significantly(P<0.05).Compared with model group,the alkaline phosphatase(ALP)value was significantly or extremely decreased in the administration group(P<0.05;P<0.01),and the blood WBC,RBC,HGB and PLT indexes were extremely significantly increased (P<0.01).The results indicated that the degree of body damage in the administration group was lower than that in the model group.     

Effect of Fermented Traditional Chinese Medicine on Preventing and Treating Colibacillosis and Immune Regulation in Broilers

This study was aimed to evaluate the efficacy of the fermented traditional Chinese medicine on treating colibacillosis and immune regulation in chickens.240 healthy 1-day-old AA White-feathered broilers were selected to carry out the control test of Escherichia coli O78 LD₅₀ and compound Chinese medicine fermentation broth on colibacillosis.The therapeutic effect was evaluated at 28 days old,and the immune organ index,antioxidant index and intestinal flora were measured at 28 and 42 days old.The results showed that the infection time of Escherichia coli was delayed from 4 to 24 h after the treatment of fermentation liquid of traditional Chinese medicine fermented by Lactobacillus plantarum and Bacillus subtilis.The effective rates of high dose group (1.0 g/mL),medium dose group (0.5 g/mL) and antibiotic group (neomycin sulfate) were 80.0%,76.7% and 80.0%,and the cure rates were 80.0%,70.0% and 70.0%,respectively.Compared with infection and antibiotic groups,the thymus index and bursa index in high dose group at 28 days old were increased by 165.6%(P<0.05),25.4%(P>0.05) and 82.6%(P<0.05),94.3% (P<0.05),respectively.The thymus index and bursa index at 42 days old were increased by 97.4%,82.0% and 43.4%,27.5% (P<0.05),respectively.The T-AOC,TP,GSH-Px and T-SOD antioxidant indexes were significant difference (P<0.05),and significantly increased the content of lactic acid bacteria in intestine and reduced the content of Escherichia coli at 28 and 42 days old (P<0.05).In conclusion,the medium dose group of fermented traditional Chinese medicine (0.5 g/mL) had obvious antibacterial effect and provided substitute for antibiotics,the high dose group of fermented traditional Chinese medicine (1.0 g/mL) was significantly better than antibiotics for broilers.     

Studies on Preparation and Application of Fuzhengjiedusan Compound Preparation Fermentated by Lactic Acid Bacteria in Broilers

This study was aimed toproduce compound preparations of Fuzhengjiedusan fermented with Lactic acid bacteria and analyze its effectson growth performancesand immune organ growth of broilers. Firstly, to identify the influence of traditional Chinese medicine on the growth of Lactic acid bacteria, the medium for the growth of Lactic acid bacteria was filled with different proportions of Astragalusmembranaceus, Radix Isatidis, Epimedium and compound Chinese herbal medicine extracts by single factor test. Secondly, to confirm the optimum medium, orthogonal test was used to optimize the culture medium of compound medicine liquid. Finally, 450 one-day-old healthy AA broilers were randomized into three groups, each group had 3 repeats: control group (antibiotics), group Ⅰ(fermented preparation), group Ⅱ (unfermented preparation). The growth and immune organ index were determined in 21 and 42 days after administration. The results showed that the highest viable count of Astragalusmembranaceus, Radix Isatidis, Epimedium and compound medicine liquid groups were increased by 261.64%, 505.48%, 156.16% and 564.38%, respectively, compared with control group. The optimum medium composition was compound medicine liquid 100 mL, brown sugar 2.0 g, peptone 0.3 g, soybean meal 2.0 g. The viable count of fermentation liquid was 7.13×10⁹ CFU/mL.The production results showed that in period of 1~42 days, the daily weight gain of group Ⅰ and Ⅱ were separately increased by 17.84% and 8.22% (P<0.05) than that of the control. What's more, the feed conversion rate was reduced by 10.88% (P<0.05). The average daily feed intake was significantly higher than control group (P<0.05). At 21 and 42 days, the spleen index of group Ⅰ were improved by 11.04% and 13.58% (P<0.05), respectively. The bursal index were raised by 31.30% and 26.23% (P<0.05), respectively. However, the spleen index of group Ⅱ were improved by 7.79% and 7.41% (P<0.05), respectively. The bursal index were raised by 15.65% and 11.48% (P<0.05), respectively. On the basis of the above results, it could be suggested that Chinese herbal medicine extracts can promote lactic acid bacteria proliferation and compound fermentation liquid can improve growth performance in broilers, as well as immune organ development.     

乳酸菌发酵扶正解毒散复合制剂的制备及其在肉鸡上的应用研究

本研究旨在制备一种乳酸菌发酵的扶正解毒散复合制剂,并探索其对肉鸡生长性能和免疫器官发育的影响。首先,利用单因子试验法将不同比例的黄芪、板蓝根、淫羊藿及复合中药提取液添加至乳酸菌的培养基中,测定中药对乳酸菌生长的影响;其次,利用正交试验对复合药液培养基进行优化,确定最佳的培养基配方;最后,选取1日龄健康AA肉仔鸡450只,随机分为3组,每组3个重复:对照组(抗生素)、试验Ⅰ组(发酵制剂)、试验Ⅱ组(未发酵制剂),分别在试验的第21、42天测定肉鸡的生长及免疫器官指标。试验结果显示,黄芪、板蓝根、淫羊藿及复合药液组的最高活菌数与对照组相比分别提高了261.64%、505.48%、156.16%、564.38%。通过正交试验得到复合发酵液最适生产培养基配方:复合药液100 mL、红糖2.0 g、蛋白胨0.3 g、豆粕2.0 g,发酵液活菌数为7.13×10⁹ CFU/mL。肉鸡生长试验结果显示,1~42日龄阶段,Ⅰ、Ⅱ组的平均日增重分别比对照组提高了17.84%和8.22% (P<0.05),料肉比分别比对照组降低了10.88%和4.66% (P<0.05),平均日采食量明显高于对照组 (P<0.05);21、42日龄时,Ⅰ、Ⅱ组脾指数分别比对照组提高了11.04%和13.58% (P<0.05)、7.79%和7.41% (P<0.05),法氏囊指数分别比对照组提高了31.30%和26.23% (P<0.05)、15.65%和11.48% (P<0.05)。以上试验结果表明中药提取液能促进乳酸菌的增殖,复合发酵液能够提高肉鸡的生长性能并促进免疫器官的发育。     

复方中药水提物对腹水综合征肉鸡肝脏TGF-β1/Smad通路的影响

【目的】 研究转化生长因子β1(transforming growth factor-β 1,TGF-β1)/Smad通路在肉鸡腹水综合征肝纤维化中的作用及复方中药水提物的防治机理。【方法】 选取健康Ross肉鸡258只,常规饲养至7日龄后,随机分为5组:空白组(C);模型组(M),低温、高脂和高蛋白饲料、高钠饮水的多因素法造模;复方中药水提物高、中、低剂量组(T1、T2、T3),在模型组基础上,每天饮水中分别添加2.0、1.0、0.5 g/kg复方中药水提物。观察临床症状和剖检变化,通过HE染色和Masson染色分别观察肉鸡肝脏组织结构变化与胶原纤维沉积,计算15、25、35、45日龄体重变化。利用实时荧光定量PCR检测各日龄肝脏中TGF-β1、转化生长因子β受体Ⅰ(transforming growth factor β receptor Ⅰ,TβRⅠ)、Smad2、Smad3和Smad7基因相对表达量;利用ELISA法检测各日龄肝脏中TGF-β1、TβRⅠ蛋白含量。【结果】 与空白组相比,模型组肉鸡腹部膨大、触压有波动感;腹腔内有大量淡黄色液体,肝脏充血肿胀、质脆,表面有瘀血点;肝脏组织结构紊乱,炎性细胞浸润,胶原纤维大量沉积,肝脏发生纤维化;体重极显著降低(P<0.01);除15日龄TβRⅠ蛋白外,肝脏中TGF-β1、TβRⅠ基因mRNA相对表达量与蛋白含量均极显著或显著升高(P<0.01;P<0.05),Smad2、Smad3基因mRNA相对表达量极显著升高(P<0.01),Smad7基因mRNA相对表达量极显著降低(P<0.01)。与模型组相比,复方中药水提物组的肉鸡上述结果均有不同程度改善,且高剂量组作用效果最显著。【结论】 肝纤维化参与了肉鸡腹水综合征的发生发展,机制与TGF-β1/Smad通路相关;由黄芪、当归、丹参、茯苓等制成的复方中药水提物可以调控TGF-β1/Smad通路,抑制肝纤维化的发生,可有效防治肉鸡腹水综合征。     

黔北麻羊SRD5A2基因干扰载体构建及其对产羔相关基因表达的影响

旨在探究SRD5A2基因对山羊产羔性状相关基因表达的影响。本研究选择健康的（2~3岁）黔北麻羊经产母羊3只,体重约32 kg,采集卵巢组织并成功培养卵巢颗粒细胞。通过在线软件设计SRD5A2基因的4对siRNA干扰序列和1对阴性对照序列,连接pGPH1/GFP/Neo载体后,将构建成功的干扰载体转染至黔北麻羊卵巢颗粒细胞。利用qRT-PCR方法筛选出干扰效率最佳的载体,检测其对SRD5A2基因表达的影响。运用qRT-PCR检测沉默SRD5A2基因对山羊产羔性状相关基因骨形态发生蛋白15（BMP15）、生长分化因子9（GDF9）、骨形态发生蛋白1B（BMPR-1B）和卵泡刺激素β亚基（FSHβ）表达的影响。结果表明,本试验在培养黔北麻羊卵巢颗粒细胞的基础上,成功构建了黔北麻羊SRD5A2基因的干扰载体,并筛选出shRNA-SRD5A2-1干扰效果最佳（P<0.01）;抑制SRD5A2基因表达后,qRT-PCR检测产羔性状相关基因BMP15、BMPR-1B、GDF9和FSHβ的表达量均显著降低,BMP15的表达量极显著低于shRNA-NC对照组（P<0.01）,BMPR-1B、GDF9和FSHβ的表达量显著低于shRNA-NC对照组（P<0.05）。本研究成功构建了SRD5A2基因干扰载体并转染至卵巢颗粒细胞,发现体外沉默SRD5A2基因可抑制产羔性状相关基因的表达,提示SRD5A2基因与黔北麻羊产羔性状密切相关,本试验结果为进一步研究SRD5A2基因对黔北麻羊产羔性状的调控机制提供了基础。