骆驼乳清蛋白对热应激所致大鼠肝氧化损伤的保护作用

本试验旨在研究骆驼乳清蛋白（CWP）对热应激（HS）大鼠肝损伤、氧化应激及肝功能的保护作用。选取6周龄SD大鼠36只,适应性饲养2周后随机分为6组：正常对照组饲喂基础日粮;CWP对照组添加400 mg·kg^-1的CWP;HS组除饲喂基础日粮外,每天进行2 h HS处理,连续8 d;3个CWP干预组分别于每次HS前灌服100、200和400 mg·kg^-1的CWP。试验结束后,取大鼠肝组织,HE染色观察肝组织病理学变化,同时检测肝功能生物标志物、氧化应激标志物水平及抗氧化酶活性。结果表明：CWP降低了HS大鼠血清谷草转氨酶（AST）和谷丙转氨酶（ALT）活性,400 mg·kg^-1的CWP干预效果最好（P<0.01）;400 mg·kg^-1CWP干预组有效降低了HS诱导的大鼠肝组织病理学改变; CWP降低了大鼠肝活性氧（ROS）及丙二醛（MDA）水平,增加了超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH-Px）活性及谷胱甘肽（GSH）水平,400 mg·kg^-1的CWP干预效果最好（P<0.01）。结果提示,CWP干预能够以剂量依赖性方式降低大鼠肝氧化应激,增加抗氧化系统防御能力,从而缓解HS所致大鼠肝损伤。     

Protection of farmed camels (Camelus Dromedarius): Welfare problems and legislative perspective

In the last years animal welfare has assumed an increasing interest in our society, influencing legislation to enact many provisions aimed at the protection of animals. Along with increased consumer awareness of the need to maintain ethically acceptable conditions of raised animals, scientists too have begun to investigate the conditions of animal welfare, the tools for its evaluation and for its improvement. Although there are many advances in knowledge, much remains to be investigated concerning many species considered "minor", that is, camels and dromedaries. Dromedaries, recently, have attracted the interest of some breeders following the results of studies concerning the nutritional and therapeutic properties of their products ‐ milk in particular ‐ that make them ideal for some particular categories of consumers, such as diabetics, obesity sufferers, lactose‐intolerant subjects, menopausal women and so on. Considering their use in dairy husbandry, dromedaries are reared under intensive and/or semi‐intensive systems with the resulting emergence of specific needs, which should be fulfilled in order to have appropriate welfare. This paper's purpose is to give practical elements in order to find out dromedary welfare standards, promoting a comprehensive set of regulations on welfare, care and protection of this animal.     

Relevance of Dried Meat Product （Kundi）, an Intermediate Moisture Meat,for Food Security

The main goal of food security is for individual to be able to obtain adequate and quality food needed at all times, and to utilise the food to meet the body needs. Kundi is a relish Intermediate Moisture Meat （dried meat） product, produced in the Northern part of Nigeria. 2-3 years old male Camelus dromedarius and White Fulani animals were used for this study, 2 kg of semimembranous muscles from each animal were used, and trimmed of all external fat, connective tissues and bones. Meat samples were cut in sizeable pieces of weight ranges 70-90 grams of 6-8 cm and kept in the refrigerator for 24 hours. Fresh meat cuts were boiled for 20 minutes at 100 ~C, seasoned and oven dried for 3 hours at 170 ~C. This study assessed the proximate composition of fresh and dried meat （Kundi） products and their sensory evaluation in a completely randomized design. The results showed that fresh camel meat had 74.55% and 21.96% significantly （P 〈 0.05） higher in moisture and protein content than 72.69% and 18.96% of fresh beef, while fresh beef had higher （P 〈 0.05） ether extract （6.34%） than fresh camel meat （2.39%）. Laboratory Prepared Beef Kundi （LPBK） had the highest （P 〈 0.05） value in moisture content 35.09% followed by Laboratory Prepared Camel ＇Kundi＇ （LPCK） with 33.21% while Commercial Kundi （CK） had the least. Protein obtained was inversely proportionally to moisture content. Ether extract for LPCK and LPBK were statistically similar （P 〉 0.05） and both were lower （P 〈 0.05） than value obtained for CK. The results obtained for sensory evaluation showed that the panelist rated seasoned Kundi to have the highest significant （P 〈 0.05） valve for tenderness, flavour, colour, juiciness, texture and acceptability with values 6.50, 5.30, 6.50, 6.53, 6.30 and 7.00 respectively. Study showed that dry meat products （Kundi） is of high nutritional qualities, and could be of a great relevance; to food security.     

家养双峰驼SRY基因的序列分析与拷贝数研究

为探究中国家养双峰驼SRY基因的序列以及群体间拷贝数变异,对7个中国家养双峰驼群体共94个个体的SRY基因部分序列进行测序,分析SRY基因序列与其他物种的相似性;并且以CYP2A基因为内参基因,运用实时荧光定量PCR技术检测SRY基因的拷贝数。结果表明,中国家养双峰驼与人、马、猪、羊及普通牛的SRY基因相似度分别为67.7%~68.1%、68.5%~69.4%、73.8%~74.4%、73.8%~74.8%、74.1%~74.8%。在家养双峰驼SRY基因的多个拷贝中,除1个拷贝外,其余拷贝的HMG序列与羊驼、人、马、猪、绵羊及普通牛的HMG序列相比,相似度分别为97.3%、84.2%、85.6%、86.3%、87.0%、85.6%。SRY基因在中国家养双峰驼Y染色体上以多拷贝形式存在,变幅为1~9个拷贝。表明中国家养双峰驼SRY基因的HMG序列是高度保守的,而且该基因属于多拷贝基因。     

一株骆驼源大肠杆菌的分离鉴定与药敏试验

新疆某骆驼园发生多例驼羔腹泻,临床表现为严重水样腹泻,患病驼羔体温升高,可高达40℃以上;表现为腹泻、脱水、败血症、心力衰竭,迅速死亡。剖检病死驼羔,有肠系膜淋巴结肿大,小肠黏膜充血。无菌采取病死驼羔的肺脏、肠等组织进行微生物学诊断,对分离菌进行革兰氏染色镜检、小鼠致病性试验和16S rDNA基因鉴定明确分离菌为大肠杆菌。后进行药敏试验,结果显示该分离大肠杆菌对替米考星、头孢曲松钠、氟苯尼考敏感。     

新疆双峰驼纳米抗体、重链抗体和常规抗体的热稳定性比较

【目的】 比较纳米抗体、HCAbs与常规抗体之间在温度稳定性方面的差异,为研究HCAb的功能特性和骆驼适应极端环境的免疫特点提供参考。【方法】 从溶菌酶、蒜氨酸酶和丹毒丝菌表面抗原A 3种抗原免疫的新疆双峰驼血清中,采用Protein A和Protein G亲和色谱纯化IgG1、IgG2和IgG3 3种亚型抗体,并在大肠杆菌BL21(DE3)菌中表达和纯化3种纳米抗体。采用ELISA方法,测定经过22~90℃一系列热处理后、平衡至室温的各抗体与相应抗原的结合活性,以剩余抗原结合活性的百分比作为衡量抗体温度稳定性的参数。【结果】 3种抗原免疫后均能在骆驼激发明显的抗体反应。采用Protein A/G亲和色谱,从血清中纯化获得分子量分别为50 KD+25 KD、 46 KD和43 KD的IgG1、IgG2和IgG3亚型抗体。骆驼IgG2和IgG3重链抗体比IgG1具有更高的温度稳定性,其中IgG3表现出比IgG2对温度更高耐受的趋势。从细菌表达纯化的3种纳米抗体都表现出比重链抗体和常规抗体更高的温度稳定性。【结论】 抗体结构形式和分子大小可能显著影响了双峰驼抗体对温度的耐受性。     

双峰驼TLR5抗体制备及其在淋巴结中的表达

【目的】制备兔抗双峰驼Toll样受体5（TLR5）的多克隆抗体并鉴定其活性。【方法】选择双峰驼TLR5基因序列,经软件分析选择其编码区的膜外区1~499氨基酸位为主要抗原位,再选取对应的核苷酸序列进行基因合成,并与pET-28a（+）载体连接构建重组质粒pET28a-TLR5,对其进行BamH Ⅰ和Xho Ⅰ双酶切鉴定。将pET28a-TLR5转染大肠杆菌原核表达系统,用IPTG进行诱导表达,对IPTG浓度（0.1,0.2,0.4,0.6,0.8,1.0 mol/L）和诱导时间（2,3,4,5,6,7,8,9 h）进行优化。将表达的蛋白经亲和层析法纯化后免疫新西兰白兔以制备多克隆抗体,采用ELISA法检测多抗效价,用Western blot检测和免疫组化验证评价多克隆抗体的特异性。【结果】成功构建了重组质粒pET28a-TLR5,经IPTG诱导后获得了重组蛋白（大小约为57 ku）,且以包涵体的形式表达。IPTG最佳诱导表达条件为：IPTG浓度为1 mol/L,诱导时间为6 h。TLR5多抗血清效价为1∶128 000。Western blot鉴定表明,TLR5多抗血清能特异性识别目的蛋白;免疫组化结果表明,TLR5多抗能很好地识别成年双峰驼十二指肠肠系膜淋巴中的TLR5阳性细胞。【结论】成功制备出特异性良好的兔抗双峰驼TLR5多克隆抗体,能够用于TLR5的检测。     

骆驼福斯盘尾丝虫病组织病理学观察

为更有效地对骆驼盘尾丝虫病进行防治,利用流行病学调查和常规病理学观察的方法,对骆驼盘尾丝虫病进行研究。结果表明:内蒙古阿拉善左旗骆驼盘尾丝虫病平均感染率为85.4%,感染强度为1~15峰-1,虫体形成的结节分布在骆驼的颈项韧带处;共收集到209个结节,它们的形态和质地各不相同,软的结节内形成纤维组织囊腔,硬的结节内形成钙化灶;骆驼年龄段不同,结节的分布呈不同变化;病理组织学观察显示,囊腔周围有各种炎性细胞侵润,囊腔内有不同状态的雌虫虫体,骆驼盘尾丝虫病表现为骆驼的寄生虫性肉芽肿。     

新疆双峰驼C3d基因cDNA的克隆与序列分析

【目的】克隆骆驼补体C3d基因,分析序列特征,为其佐剂效应研究提供依据。【方法】利用Trizol试剂,从骆驼肝脏组织提取总RNA,并反转录获得cDNA。设计C3d特异引物,应用PCR技术扩增C3d序列,构建到pMD18-T载体。将重组质粒转入感受态菌株并双酶切鉴定。使用ClustW,DNAstar, Swiss-Model软件对构建的重组C3d基因序列进行同源性比对,建立系统进化树并对其二级和三级结构进行预测和分析。【结果】采用骆驼C3d特异引物,PCR扩增获得大小为909 bp的骆驼补体C3d基因。构建至T载体并转入大肠杆菌,获得阳性转化克隆。克隆的新疆双峰驼C3d序列与骆驼科C3d基因之间的同源性在97%以上,与牛和猪C3d基因同源性为88%,与兔和人的C3d基因同源性为83%。对骆驼C3d蛋白采用Swiss-Model软件进行序列分析,预测该蛋白质的二级结构主要由α-螺旋和β-折叠组成,该蛋白可能具有良好的免疫原性和免疫结合活性。【结论】C3d基因在进化中较为保守,在骆驼免疫中可以采用亲缘关系较近的其他动物来源的C3d作为分子佐剂从而增强骆驼免疫效果。     

骆驼斯氏副柔线虫病间接ELISA检测方法的建立

为建立骆驼斯氏副柔线虫病间接ELISA （iELISA）诊断方法,本试验对骆驼斯氏副柔线虫半胱氨酸蛋白酶CPR基因进行重组表达,将获取的重组蛋白（rCPR）进行纯化和Western blotting检测,然后以纯化好的重组蛋白为抗原,通过棋盘滴定试验优化了抗原包被浓度、包被条件、抗体稀释度、酶标二抗稀释度、封闭液和封闭时间等,建立了骆驼斯氏副柔线虫病iELISA诊断方法,并对建立的iELISA检测方法进行了重复性、敏感性、特异性试验和临床检测。结果显示,抗原最佳包被浓度为8 μg/孔,血清最佳稀释度为1：50,酶标二抗最佳稀释度为1：5 000,最佳包被条件为4℃包被过夜,最佳封闭条件为3% BSA封闭2 h。临界值为0.235,待检血清D_{450 nm}值>0.235则确定为阳性。重复性试验中变异系数均<10%,重复性较好;用该方法检测阳性血清的敏感性为96.3%;此方法仅与骆驼斯氏副柔线虫病阳性血清发生特异性反应,与感染了其他寄生虫的阳性血清无交叉反应,特异性为100%;对140份临床血清进行检测,阳性率为86.4%。综上可知,本试验成功建立了一种快速有效诊断骆驼斯氏副柔线虫病的iELISA方法。