家蚕细胞色素P450基因CYP9A22的克隆及在中肠和脂肪体的诱导转录

细胞色素P450对昆虫的生长发育、环境适应性以及抗药性具有重要作用。为了探讨氯氰菊酯对家蚕(Bombyxmori)细胞色素P450基因转录的诱导作用,采用RT-PCR技术和cDNA末端快速扩增(RACE)策略,从家蚕5龄幼虫克隆到细胞色素P450基因CYP9A22(GenBank登录号:EF535804)。CYP9A22基因的cDNA编码区长1 596 bp,编码531个氨基酸,推定的蛋白质分子质量为61 kD,等电点为7.71。将该基因的cDNA和家蚕基因组序列比对,有9个内含子;与已知的CYP9家族的棉铃虫(Helicoverpa armigera)CYP9A14基因的相应氨基酸同源性达到62.3%。用半定量RT-PCR方法分析氯氰菊酯诱导家蚕CYP9A22表达水平,结果表明:CYP9A22的mRNA转录表达具有组织特异性,在中肠的表达量高于脂肪体;氯氰菊酯诱导家蚕24 h,在其脂肪体的表达量是未诱导家蚕脂肪体的3.1倍,初步推测CYP9A22的过量表达可能与抗药性有一定关系。     

茧层量差异家蚕品种后部丝腺细胞和血液组织的蛋白质表达谱分析

为了研究家蚕茧层量性状的分子调控机制,采用蛋白质双向电泳和图像分析技术,分析了遗传背景基本相同,而结茧性状和茧层量具有明显差异的正常茧、薄皮茧、裸蛹3个家蚕品种的丝腺细胞和血液组织的蛋白质表达与茧层量的关系。结果表明:裸蛹品种的丝腺细胞蛋白斑点与正常茧品种的蛋白斑点的匹配率仅在65%左右,且存在35%左右的特异蛋白斑点,暗示裸蛹性状可能是主效基因控制的微效多基因遗传;3个品种后部丝腺细胞特异蛋白斑点、匹配蛋白斑点按等电点划分的分布规律具有明显差异,而血液组织相应的分布规律非常相似,暗示丝腺细胞中的蛋白及其基因对家蚕茧层量性状的作用要大于血液组织中的蛋白及其基因;正常茧、薄皮茧和裸蛹3个品种后部丝腺细胞中特异蛋白斑点及上调和下调蛋白斑点中,可能分别存在着与家蚕高茧层量性状、结薄皮茧性状以及不结茧性状有关的蛋白质。     

Effect of Fagopyrum dibotrys (D.Don) Hara on Growth Performance,Immune Function and Intestinal Structure of Broilers

This experiment was conducted to investigate the effects of Fagopyrum dibotrys (D.Don) Hara on growth performance,immune function and intestinal structure of broilers. A total of 270 one-day-old healthy Tiejiaoma broilers were chosen and randomly divided into 3 groups with 6 replicates per group and 15 broilers per replicate.Control group was fed with the basal diet,group Ⅱ was fed with basal diet adding 8% flavomycin 60 mg/kg +15% aureomycin 270 mg/kg and group Ⅲ was fed with basal diet adding 1% Fagopyrum dibotrys (D.Don) Hara. The trial period lasted for 21 days. The results showed as follow:① At 21 days of age,the average daily feed intake of group Ⅲ was significantly lower than that of groups Ⅰ and Ⅱ (P<0.05).However,there was no significant difference in the average daily gain and F/G among the three groups (P>0.05).②At 21 days of age,the thymus and bursa of Fabricius indexes of groups Ⅱ and Ⅲ were significantly higher than that of group Ⅰ (P<0.05);But the spleen index were not significantly different among the three groups (P>0.05).③ At 21 days of age,the content of IgA and IgM in serum of groups Ⅲ were significantly higher than that of group Ⅰ (P<0.05),and the content of IgG in serum of groups Ⅱ and Ⅲ were significantly higher than that of group Ⅰ (P<0.05).④At 21 days of age,the content of IL-2,TNF-α and sIgA in intestinal mucosa of groups Ⅱ and Ⅲ were significantly higher than that of group Ⅰ (P<0.05),and the content of IL-6 in intestinal mucosa of group Ⅲ were significantly higher than that of groups Ⅰ and Ⅱ (P<0.05).⑤At 21 days of age,the villus height of jejunum in groups Ⅱ and Ⅲ were significantly higher than that of group Ⅰ (P<0.05),the villus height/crypt depth of jejunum in group Ⅱ was significantly higher than that of groups Ⅰ and Ⅲ (P<0.05). However,the crypt depth was not significantly different among the three groups (P>0.05).In conclusion,that Fagopyrum dibotrys (D.Don) Hara could not affect the growth performance of broilers,but it could improve the development of immune organs,increase the secretion of serum immunoglobulin, promote the immune function of intestinal mucosa and intestinal morphology of jejunum.     

母鸡饲粮添加肌苷酸对种蛋品质、1～21日龄子代肉鸡生长性能和血清生化指标的影响

本试验旨在研究母鸡饲粮中添加不同水平肌苷酸(IMP)对种蛋品质、1~21日龄子代肉鸡生长性能和血清生化指标的影响。选取20周龄健康爱拔益加(Arbor Acres)肉鸡480只,随机分为对照组(基础饲粮)、Ⅰ组(基础饲粮+0.2%肌苷酸)、Ⅱ组(基础饲粮+0.5%肌苷酸)和Ⅲ组(基础饲粮+1.0%肌苷酸)。出壳后的子代肉鸡随机选取480只,根据其母鸡饲粮中肌苷酸添加情况相应分为对照组、Ⅰ组、Ⅱ组和Ⅲ组,每组6个重复,每个重复20只鸡。试验期21 d。结果表明:与对照组相比,各试验组32周龄末母鸡的种蛋品质指标、1~21日龄子代肉鸡的生长性能指标和血清蛋白质代谢及酶活性相关指标均无显著差异(P0.05)。与对照组相比,Ⅲ组21日龄子代肉鸡的血清高密度脂蛋白胆固醇(HLD-C)含量显著升高(P0.05),各试验组的血清总胆固醇(T-CHO)、甘油三酯(TG)和低密度脂蛋白胆固醇(LDL-C)含量无显著差异(P0.05)。与对照组相比,Ⅱ组和Ⅲ组21日龄子代肉鸡的血清三碘甲状腺原氨酸(T_3)含量显著升高(P0.05),Ⅱ组的血清甲状腺素(T4)和胰岛素样生长因子-1(IGF-1)含量显著升高(P0.05),各试验组的血清生长激素(GH)含量无显著差异(P0.05)。与对照组相比,Ⅲ组21日龄子代肉鸡的血清免疫球蛋白M(Ig M)、白细胞介素2(IL-2)和白细胞介素6(IL-6)含量有上升趋势(P0.05)。以上结果表明,母鸡饲粮添加肌苷酸增强了子代肉鸡体内的脂质代谢,提高了子代肉鸡血清生长与免疫相关激素水平和免疫因子含量;综合种蛋品质、生长性能及血清生化指标,母鸡饲粮添加0.5%~1.0%肌苷酸效果最佳。     

基于体外细胞模型的霉菌毒素毒性评价

霉菌毒素是由多种真菌产生的次级代谢产物,广泛存在于食品和饲料中。霉菌毒素污染的粮食和饲料会给畜牧业生产和畜产品质量安全带来极大隐患。研究霉菌毒素致毒机理,为今后研究其对动物及人的影响开展更深入更全面的研究提供理论依据。细胞模型作为一种常用的体外试验方法广泛用于毒理学研究中。本文简述了黄曲霉毒素B_1(AFB_1)、赭曲霉毒素A(OTA)、脱氧雪腐镰刀菌烯醇(DON)、玉米赤霉烯酮(ZEA)和展青霉素(PAT)的一般特性,并综述了利用细胞模型进行AFB_1、OTA、DON、ZEA和PAT毒性、联合毒性及致毒机理研究的进展。     

Advances on Detection Methods for Residues of Nitrofurans in Food

Nitrofurans are broad-spectrum antibiotics which have been widely used in food animals to the prevention and treatment of gastrointestinal infections.Due to abuse of drugs by criminals,animals could produce drug-resistant strains,which reduces the role of antibiotics.Meanwhile,excessive drugs residues in food has carcinogenic,teratogenic and mutagenic effects to the human body.In this paper,the damage and the detection methods of the residues of the nitrofurans in food are surveyed.The main detection methods include high performance liquid chromatography,liquid chromatography-mass spectrometry,liquid chromatography tandem mass spectrometry,enzyme-linked immunoassay,colloidal gold immunoassay and fluorescence immunity analysis method.The research status of various detection methods are summarized and the suggestions for further research are put forward.It is expected to provide reference for future research.     

基于《耕地质量等级》的靖远县耕地质量等级评价

基于《耕地质量等级》标准,按照黄土高原区耕地质量等级评定指标体系,利用县域耕地资源管理信息系统建立耕地质量等级信息系统数据库,运用空间分析、层次分析、模糊数学和综合指数等方法对靖远县耕地质量等级进行综合评价.结果表明:靖远县耕地受地貌、地形和水文地质条件影响等级高低出现了明显的区域分布规律,从北到南,依次为十等地到三等地,面积占比分别为1.51％、23.51％、41.55％、15.92％、10.31％、3.87％、2.34％、0.99％.耕地质量平均等级为7.64等,耕地主要利用类型由旱地和砂田逐渐向菜地和水田过渡.全县耕地土壤有机质、有效磷、速效钾平均含量为11.41 g/kg、16.55 mg/kg、178.03 mg/kg,均属于中等水平.全氮为0.70 g/kg,属于低等水平.该评价较准确地反映了靖远县耕地质量等级现状,可对耕地质量的监测和保护提供了参考依据.     

肯塔基沙门菌对氟喹诺酮和碳青霉烯类药物耐药性研究进展

肯塔基沙门菌（Salmonella Kentucky）是继鼠伤寒沙门菌和肠炎沙门菌后又一与人类肠道疾病相关的沙门菌血清型,因其获得和传播耐药性的能力较强而逐渐引起关注。该血清型沙门菌按照多位点序列分型分得的众多亚型中,ST198血清亚型具有易获得耐药突变的基因组岛的特性,21世纪以来不断进化,除获得常见抗菌药物耐药基因外,还相继获得对氟喹诺酮类和碳青霉烯类药物的耐药性并扩散全球,造成临床上难以治疗的病例而引起世界卫生组织的关注。作者整理了肯塔基沙门菌的进化历程,着重阐述肯塔基沙门菌关于氟喹诺酮类及碳青霉烯类药物的耐药情况及耐药机制,以期为中国沙门菌的实验室防控监测系统提供理论依据。     

猪RUVBL2真核表达载体的构建及其在CHO细胞中的表达

RUVBL2(RuvB-like 2)蛋白是进化上高度保守AAA+(ATPases Associated With Diverse Cellular Activities,AAA)家族成员之一,CHO(Chinese Hamster Ovary)细胞被广泛地用于表达重组DNA蛋白。为探究猪睾丸组织的RUVBL2基因是否能够在CHO细胞中表达,本实验以猪睾丸组织的cDNA为模板,PCR扩增RUVBL2目的基因,并将其克隆至pIRES2-EGFP(Mammalian Expression Vectors pIRES2-EGFP)载体上,进一步转化到DH5α中,再进行PCR、酶切及测序鉴定;将重组质粒转染到CHO细胞中,再进行荧光、RT-PCR、Western blot检测。结果显示:PCR、酶切及测序都证实了RUVBL2基因正确地插入到了载体质粒pIRES2-EGFP的多克隆位点;荧光、RT-PCR、Western blot也证实了RUVBL2基因在CHO细胞中的成功表达。本实验成功构建了猪的pIRES2-EGFP-RUVBL2真核表达载体,并证实了猪睾丸组织中的RUVBL2基因能够在CHO细胞中表达。     

江西地方品种水牛产肉性能及肉品质分析研究

对18～24月龄鄱阳湖、峡江和信丰山地3个江西地方品种水牛各3头,进行肉用性能和牛肉主要营养养分的测定分析。结果表明,水牛肌肉肉色符合标准,且无PSE肉、DFD肉产生,符合优质肉牛标准;鄱阳湖水牛的胴体重、净肉重、胴体产肉率、Fe含量、Zn含量较高,显著高于信丰山地水牛（P<0.05）;鄱阳湖水牛18种氨基酸含量除酪氨酸外均高于其它2个地方品种,其中天门冬氨酸、谷氨酸、丝氨酸、脯氨酸、丙氨酸、亮氨酸及胱氨酸含量显著高于信丰山地水牛（P<0.05）,鲜味氨基酸、甜鲜味氨基酸含量显著高于信丰山地水牛（P<0.05）;牛肉脂肪酸中花生酸含量在品种间存在显著差异（P<0.05）,而其它各种脂肪酸含量、硬脂肪酸总含量和不饱和脂肪总含量在品种间均无显著差异（P>0.05）。