Effects of dietary cecropin on growth,non‐specific immunity and disease resistance of tilapia (Oreochromis niloticus × O. aureus)

An 8‐week feeding trial was conducted to evaluate the effect of dietary cecropin on growth performance, non‐specific immunity and disease resistance of hybrid tilapia (Oreochromis niloticus × O. aureus). A basal diet was supplemented with 0 (control), 75, 150 and 225 mg kg^?1 cecropin to formulate four experimental diets. The results showed that final body weight and specific growth rate obtained with diet 150 mg kg^?1 cecropin were significantly higher (P < 0.05) than that obtained with diets 0 and 75 mg kg^?1, whereas there was no difference between 150 and 225 mg kg^?1. Feed conversion efficiency was improved by increasing the dietary cecropin content (P < 0.05). However, feeding intake was reduced as dietary cecropin increased. Serum lysozyme, alkaline phosphatase, catalase and superoxide dismutase activity were significantly increased with the increased levels of dietary cecropin (P < 0.05), and reached a maximum at level of 150 mg kg^?1 cecropin, then those values no longer increased. Moreover, the dietary cecropin supplementation level also exhibited a decrease in the cumulative mortality rates compared with the controls when challenged with Aeromonas veronii. The results suggested that the dietary supplementation of cecropin could improve the growth, non‐specific immunity indicators and resistance of hybrid tilapia against A. veronii.     

HTPP-MDC inhibits replication and subcellular localization of hepatitis B virus in HepG2.2.15 cells

AIM：To investigate the antiviral effect of hepatocyte-targeting and cell-penetrating peptide and Musca domestica cecropin (HTPP-MDC) fusion polypeptide against hepatitis B virus (HBV) in vitro, and to observe the penetrating ability of HTPP-MDC in hepatocytes. METHODS： HepG2.2.15 cells and Chang liver cells were co-cultured in vitro with HTPP-MDC at different concentrations. MTT assay was used to detect the cytotoxicity of HTPP-MDC in vitro. The levels of HBsAg, HBeAg and HBV DNA in HepG2.2.15 cell culture supernatants were measured by ELISA and real-time fluorescence quantitative PCR. The penetrating ability of HTPP-MDC was detected by laser confocal microscopy. RESULTS：The levels of HBsAg, HBeAg and HBV DNA in the supernatants of HepG2.2.15 cells treated with HTPP-MDC were remarkably reduced. The inhibitory effect of HTPP-MDC depended on the dose and action time of the drug. FITC-labeled HTPP-MDC was observed inside the cells under laser confocal microscope. CONCLUSION：HTPP-MDC strongly inhibits HBV replication in HepG2.2.15 cells. The penetrating ability of HTPP-MDC into hepatocytes indicates that HTPP-MDC is useful in clinic therapy for chronic hepatitis B-related diseases in the future.     

双抗植物表达载体的构建

用限制性内切酶将抗虫的胰蛋白酶抑制剂（ＴＩ）基因从质粒ｐＢｉｎ２０上切下,插入带有抗细菌的抗菌肽Ｂ和抗菌肽Ｄ基因的植物表达载体ｐＤＢ３的ＸｂａⅠ位点。通过点杂交和酶切鉴定,得到了带有３个抗性基因的双抗植物表达载体ｐＤＢＴ．     

天蚕素和合生素对AA肉鸡小肠黏膜形态和小肠免疫细胞的影响

为研究日粮中添加天蚕素、合生素对AA肉鸡小肠黏膜形态及免疫细胞的影响,试验选用480只1日龄健康AA肉鸡,随机分为4组,每组4个重复,每个重复30只鸡。对照组（Ⅰ组）饲喂基础日粮,试验组（Ⅱ~Ⅳ组）分别在基础日粮中添加0.5%天蚕素、0.3%合生素、0.5%天蚕素+0.3%合生素。试验期42 d。结果显示,与Ⅰ组相比,Ⅳ组十二指肠、回肠绒毛高度极显著升高（P<0.01）;Ⅱ组空肠和回肠、Ⅲ组小肠各段隐窝深度显著或极显著降低（P<0.05;P<0.01）;Ⅱ、Ⅲ组小肠各段和Ⅳ组十二指肠、回肠绒毛高度/隐窝深度显著或极显著升高（P<0.05;P<0.01）;Ⅱ组空肠绒毛宽度显著降低（P<0.05）,Ⅲ组十二指肠和空肠、Ⅳ组空肠和回肠绒毛宽度均极显著降低（P<0.01）;Ⅱ、Ⅲ组空肠黏膜厚度显著升高（P<0.05）,Ⅳ组小肠各段黏膜厚度均极显著升高（P<0.01）。Ⅱ组回肠上皮内淋巴细胞数量显著升高（P<0.05）,Ⅲ组十二指肠、Ⅳ组小肠各段上皮内淋巴细胞数量极显著升高（P<0.01）;Ⅲ组十二指肠、Ⅳ组十二指肠和回肠杯状细胞数量极显著升高（P<0.01）;Ⅳ组空肠肥大细胞数量显著升高（P<0.05）。综上所述,天蚕素、合生素单独或联合添加均能改善AA肉鸡小肠黏膜结构,促进小肠黏膜免疫细胞增殖,联合添加效果最佳。     

几丁质酶和抗菌肽D双价基因转化茄子的研究

在建立茄子的组织培养及遗传转化体系的基础上，以根癌农杆菌为介导，采用叶盘法将几丁质酶和抗菌肽D双价基因转入茄子，分别获得6株Kan^R植株。对Kan^R植株进行点杂交、PCR扩增等分子检测。结果表明，目的基因已整合进茄子核基因组中。     

天蚕素A-马盖宁杂合肽对小鼠小肠黏膜结构、小肠黏膜免疫功能和肠道菌群的影响

本试验旨在研究天蚕素A-马盖宁杂合肽对小鼠小肠黏膜结构、黏膜免疫功能和肠道菌群的影响。选取18只健康且体重相近的BALB/c小鼠随机分成3组:对照组(生理盐水0.75 m L/d灌胃)、低剂量杂合肽组(0.26 mg/m L的杂合肽0.75 m L/d灌胃)、高剂量杂合肽组(0.52 mg/m L的杂合肽0.75 m L/d灌胃)。试验期为6周。结果表明:1)2个杂合肽组十二指肠绒毛长度均显著大于对照组(P<0.05),各段小肠的隐窝深度均显著低于对照组(P<0.05),各段小肠绒毛长度/隐窝深度均显著高于对照组(P<0.05)。2)与对照组相比,2个杂合肽组各段小肠黏膜内免疫球蛋白A阳性表达水平均显著升高(P<0.05)。3)2个杂合肽组小肠黏膜内白细胞介素(IL)-2、干扰素-γ(IFN-γ)及IL-4含量均显著高于对照组(P<0.05),而IFN-γ/IL-4各组之间差异不显著(P>0.05)。4)2个杂合肽组盲肠内容物中的大肠杆菌数量均显著低于对照组(P<0.05),双歧杆菌与乳酸杆菌数量均显著高于对照组(P<0.05)。由此得出,天蚕素A-马盖宁杂合肽灌胃能改善机体小肠黏膜结构;可促进免疫球蛋白A的表达来提高小肠黏膜免疫防御功能;可促进IL-2及IFN-γ的分泌来提高肠道细胞免疫水平,促进IL-4的分泌以提高肠道体液免疫水平并能够保持辅助性T细胞(Th)1/Th2的平衡状态;可有效降低肠道致病菌大肠杆菌的数量并显著增加肠道益生菌双歧杆菌和乳酸杆菌的数量。     

天蚕素抗菌肽对肉兔生产性能和免疫器官指数的影响

试验旨在研究天蚕素抗菌肽对新西兰兔生产性能和免疫器官指数的影响。选择160只35日龄断奶的新西兰肉兔,随机分为5个处理,每个处理4个重复,每个重复8只兔。对照组饲喂基础饲粮,试验组分别在基础饲粮基础上添加30 mg/kg喹乙醇、150、200 mg/kg和250 mg/kg天蚕素抗菌肽,试验期为8周。结果表明,天蚕素抗菌肽可一定程度提高肉兔的日增重,显著降低料重比(P<0.05),对肉兔的采食量无显著影响(P>0.05);添加天蚕素抗菌肽有增加肉兔脾脏和胸腺重量的趋势,添加200 mg/kg和250 mg/kg天蚕素抗菌肽可显著提高肉兔脾脏指数和胸腺指数(P<0.05)。由此可见,在肉兔饲粮中添加天蚕素抗菌肽,可提高肉兔生产性能,促进免疫器官发育,提高免疫器官指数。     

天蚕素AD和蛙Buforin Ⅱ融合蛋白在大肠杆菌中的表达

本试验采用改良二步全基因合成法人工合成了含大肠杆菌偏爱密码子的天蚕素AD和蛙Buforin Ⅱ成熟片段的融合基因,并克隆至pMD18-T中,经测序确认正确后,与pET-Trx连接,构建了表达载体pET-Trx-CAD-Buforin Ⅱ,并在大肠杆菌BL21(DE3)中诱导表达。表达产物占菌体总蛋白的35%,经羟胺裂解后对测试菌具有较好的杀菌效果,为进一步的开发与应用打下了基础。