牙鲆血清免疫球蛋白的分离纯化及部分特性分析

运用Sephacryl S-200凝胶层析和HiTrap rProtein ASepharose亲和层析2种方法对牙鲆(Paralichthys olivaceus)血清免疫球蛋白进行分离纯化,结果表明,牙鲆免疫球蛋白分布于33%~50%的硫酸铵饱和溶液中,其中45%的分离效果最好。凝胶层析和亲和层析样品均出现2个蛋白峰,用还原SDS-PAGE检测确定牙鲆免疫球蛋白存在于第2个蛋白峰中。牙鲆免疫球蛋白重链分子量约为75.4 kD,轻链分子量约为29.9 kD和28.2 kD,推测牙鲆血清免疫球蛋白的分子量为836 kD。制备了兔抗牙鲆免疫球蛋白多克隆抗体,免疫双扩散法检测多克隆抗体效价为1∶32,免疫斑点法检测多克隆抗体效价至少为1∶1 600。运用免疫印迹法(Western-bloting)检测了兔抗牙鲆免疫球蛋白多克隆抗体的特异性,实验证明该抗体与牙鲆全血清中免疫球蛋白重链、轻链反应均成阳性。     

鲤胆汁与血清中免疫球蛋白的比较研究

以正常生活状态下的非免疫鲤为材料，采用盐析，高速离心，柱层析等方法，从鲤胆汁与血清中分离提取了免疫球蛋白，并对它们的理化性质和免疫原性进行了比较研究。结果表明：鲤胆汁中存在的免疫球蛋白的某些理化性质同血清中免疫球蛋白的相似，并且二者具有相同的免疫原性。由此推断，鲤胆汁中免疫球蛋白的来源可能与血清中免疫球蛋白具有一定的同源性。     

黄鳝皮肤黏液抗菌肽的分离纯化及其部分特性研究

以黄鳝皮肤黏液为材料,通过Sephadex G-50凝胶层析和DEAE Sepharose Fast Flow离子交换层析,分离纯化了具有抗菌和镇痛活性的多肽MAS-Ⅳ-4,用Tricine-SDS-PAGE测得其分子量为10.4 kD,用显微熔点仪测得其熔点为208～211℃.用抑菌圈法测定其抑菌活性,结果表明,MAS-Ⅳ-4对嗜水气单胞菌、金黄色葡萄球菌、枯草芽孢杆菌有较强的抑菌效果.此外,每只注射0.75 ml以上的MAS-Ⅳ-4样品液能显著提高小白鼠的痛阈值.MAS-Ⅳ-4还具有微弱的溶血活性和丝氨酸蛋白酶抑制活性.     

黄鳍棘鲷血清IgM的纯化及兔抗血清的制备

分别采用Sepharose-6B凝胶过滤层析、Phenyl Sepharose FF疏水作用层析和rProtein A Sepharose亲和层析技术纯化黄鳍棘鲷（Acanthopagrus latus）血清IgM，并将这3种层析纯化方法进行了比较。纯化产物进行SDS—PAGE，电泳结果扫描后经Band Scan5．0软件分析显示，单独使用Sepharose-6B凝胶过滤层析或Phenyl Sepharose FF疏水作用层析得到的IgM纯度只有56％左右；二者联合使用纯度可以达到69％，而rProtein A Sepharose亲和层析一步就可以达到89％的纯度；纯化得到的黄鳍棘鲷血清IgM的重链和轻链分子量分别为73．6kD和26．5kD。本研究还以rProtein A Sepharose亲和层析纯化的黄鳍棘鲷血清IgM为抗原，制备其兔抗血清，采用Western Blot和双向琼脂扩散检测抗血清免疫学活性，并用间接ELISA检测其效价达到1：25600，为进一步开展黄鳍棘鲷免疫学方面的研究奠定了基础。     

固始鸡血清免疫球蛋白G的纯化及分子量的测定

采用硫酸铵分级沉淀法、葡聚糖凝胶Sephadex-25层析脱盐、DEAE-纤维素纯化等方法获得固始鸡血清免疫球蛋白G,再用SDS-聚丙烯酰胺凝胶电泳法进行分离,发现固始鸡血清免疫球蛋白显示两条蛋白谱带,它们分别是固始鸡血清免疫球蛋白G的重链(H)和轻链(L),与已知分子量的标准参照蛋白比较,测得其分子量分别约为72000kD、30000kD。     

奥尼罗非鱼血清免疫球蛋白的纯化及分子量的初步分析

通过饱和硫酸铵盐析结合 Sephadex G-200柱层析的方法,纯化制备了健康非免疫状态下奥尼罗非鱼的免疫球蛋白,进行变性还原条件下的聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹试验对血清的免疫球蛋白进行初步分析,发现SDS-PAGE电泳条件下血清重链分子量为85 kD,轻链分子量为30 kD.如果罗非鱼血清免疫球蛋白在自然状态与其他硬骨鱼类一样也为四聚体,那么其总分子量的理论值应为920 kD.     

温度对牙鲆皮肤黏液抗体产生的影响

在9℃、15℃、21℃和26℃4种不同水温下,用淋巴囊肿病毒(LCDV)灭活疫苗腹腔注射牙鲆,应用间接酶联免疫吸附试验(ELISA)研究了其皮肤黏液中特异性抗体水平的变化,以分析温度对牙鲆皮肤黏液抗体产生的影响.ELISA结果表明,9℃和15℃水温下牙鲆黏液OD值分别在注射LCDV后第9周和第7周达到峰值(9℃:OD=0.179; 15℃:OD=0.233); 21℃水温下OD值上升最快,5周达到峰值(0.316); 26℃水温下OD值较21℃无显著差异,也于5周达到峰值(0.295).采用硫酸铵分步盐析等技术粗提不同温度下OD值最高时的牙鲆皮肤黏液中的免疫球蛋白(Ig),SDS-PAGE检测发现,各温度组黏液蛋白中均含有72 kD和26 kD蛋白条带.Western blotting结果显示,抗牙鲆血清Ig重链的单克隆抗体只与黏液蛋白中72 kD条带发生反应,确定为牙鲆皮肤黏液Ig重链.综上结果表明,牙鲆在最适生活温度(21℃)下,抗体应答强度最大.牙鲆粗提黏液蛋白中Ig的初步确定,为探索牙鲆黏液免疫机制提供了材料.     

黄鳝血清白蛋白的纯化及抗体制备

通过阴离子交换层析和凝胶过滤层析,从黄鳝血清中得到其血清白蛋白,命名为MAA-serum,分子量为67kDa,为单链非糖蛋白,其N末端氨基酸序列为GHKKW。用0.5mg/只MAA-serum作为免疫源免疫大白兔,经3次免疫后,抽取大白兔血液,分离血清,饱和硫酸氨沉淀,获得抗MAA-serum的多克隆抗体。经ELISA方法测定该抗体与黄鳝血清白蛋白的免疫反应,抗体滴度为1∶100000。经初步研究发现黄鳝皮肤中也有大量血清白蛋白分布,该抗体为进一步研究MAA-serum在皮肤中的分布及生物学功能打下基础。     

不同规格黄鳝消化酶活性的比较

以相同环境条件下饲养2个月的黄鳝(Monopterus albus)为试验对象,以酶学方法研究3种规格[(143.81±36.27)g、(76.63 ±14.43)g、(31.35 ±5.82)g]黄鳝的肝脏、前肠、后肠蛋白酶、脂肪酶和淀粉酶的活性以及消化酶(蛋白酶、脂肪酶和淀粉酶)在肝脏、前肠、后肠中的活性分布.结果表明,黄鳝蛋白酶活性从高到低依次为前肠＞后肠＞肝脏；淀粉酶活性从高到低依次为后肠＞前肠＞肝脏；脂肪酶活性从高到低依次为后肠＞前肠＞肝脏；随着黄鳝体重的增加,肠道蛋白酶活性逐渐升高,而肝脏蛋白酶活性变化不明显；后肠、肝脏淀粉酶活性随着黄鳝体重的增加而升高,而前肠淀粉酶活性降低；黄鳝脂肪酶活性有随着规格变大先升高、后降低的趋势,以中规格黄鳝消化组织的脂肪酶活性最高.     

黄鳝内脏铁型超氧化物歧化酶的纯化

经丙酮分级沉淀,DEAE-琼脂糖离子交换层析和Sephacry1S-200凝胶过滤,从黄鳝内脏中分离纯化获得铁超氧化物歧化酶(Fe-SOD),并对其部分性质进行分析鉴定。获得该酶的比活力为1500U mg,提纯倍数为368 5,回收率为24 7%。该酶最大紫外吸收波长为280nm。聚丙烯酰胺凝胶电泳和等电点聚焦电泳结果表明纯化酶蛋白呈一条带,测得该酶分子量约为87kD,亚基分子量约为14 5kD;等电点为pH7 1。     

Achieving sustainable aquaculture: Historical and current perspectives and future needs and challenges

Important operational changes that have gradually been assimilated and new approaches that are developing as part of the movement toward sustainable intensive aquaculture production systems are presented via historical, current, and future perspectives. Improved environmental and economic sustainability based on increased efficiency of production continues to be realized. As a result, aquaculture continues to reduce its carbon footprint through reduced greenhouse gas emissions. Reduced use of freshwater and land resources per unit of production, improved feed management practices as well as increased knowledge of nutrient requirements, effective feed ingredients and additives, domestication of species, and new farming practices are now being applied or evaluated. Successful expansion into culture of marine species, both off and on shore, offers the potential of substantial increases in sustainable intensive aquaculture production combined with integrative efforts to increase efficiency will principally contribute to satisfying the increasing global demand for protein and food security needs.     

Fish assemblage associations and thresholds with existing and projected oil and gas development

Energy development threatens fish and wildlife resources worldwide. This study used constrained ordinations to show fish assemblage structure associated with oil and gas well densities in the Colorado River Basin, Wyoming, but well densities explained only 6.4% of assemblage structure when compared to other factors. Threshold Indicator Taxonomic ANalysis showed significant negative threshold responses by some species to small levels of development (<0.15 wells km^?2), whereas positive thresholds were less distinct. Some native and imperilled species could be disproportionately affected if future oil and gas development proceeds in a manner that will impact aquatic resources. Although existing development has not substantially influenced regional fish assemblage structure, it appears to affect a subset of species. Understanding assemblage‐level responses to development can help land managers determine appropriate development levels, prioritise areas for monitoring associated with future development and identify where land protection measures may be needed to offset potential risks.     

Gar biology and culture: status and prospects

Many lepisosteid populations in North America have declined and many are now threatened as a consequence of habitat loss and alteration and commercial and sport overfishing. Over the last two decades, morphological, histological and molecular studies allowed distinguishing between different phases of development and the nutritional condition of larvae. Ontogeny of the digestive enzymes of gar larvae indicated the possibility to feed them artificial feeds since early developmental stages. An in vitro digestibility system to test different feed ingredients has been used. Important characteristics of artificial diets were identified through different feeding experiments. Endocrinological studies showed the feasibility of altering larval development and the digestive capacity of larvae. Cloning of gar growth hormone opened new avenues to enhance growth in the gars. Plasmatic vitellogenin was isolated and purified, to develop a competitive enzyme‐linked immunosorbent assay, which allowed the straightforward separation of males from females to establish appropriate proportions for reproduction and also was used to evaluate hormonal protocols to induce gonad recrudescence and spawning. This review analyzes the biology, ecology and physiology of different gar species as a basis for their domestication, mass production of larvae for repopulation experiments and for the culture of commercial‐size gar.     

Esophagoscopy and gastroscopy in dogs and cats. Technics and indications

A report is presented of esophagoscopy and gastroscopy in cats and dogs using flexible endoscopes. Following consideration of the basic equipment, an account is given of patient preparation and anesthesia, topography, investigation technique and normal findings. Diagnostic potential, indications and risks involved are discussed.     

加快城乡统筹，推进我国农垦体制创新与经济发展的研究与思考

“十一五”以来，我国农垦加快体制机制改革、结构调整和经济增长方式的转变的步伐，参与市场竞争，努力提升企业和产品竞争力，经济社会事业发展较快，对国家的贡献提升，职工收入、生活质量明显提高和改善，示范引领作用不断增强。本文针对农垦体制机制创新进展滞缓，战略定位与发展方向不明，经济增长方式转变和结构调整缺乏资本、人才、技术的支撑，没有处理好改革、发展、稳定、民生的关系，投资匮乏，发展后劲不足，人才短缺和劳动力素质低下的现状，提出在未来农垦体制机制创新中要特别关注改革方向、目标设定与评估，改革要在现行法律法规的框架内运作，改革时机的选择，处理好改革发展稳定与民生的关系，调整好内部利益分配关系，自觉融入区域经济，与地方协同发展等问题。为了促进农垦的发展，建议国家加大对农垦体制机制创新、现代农业建设、国有农场土地资源的保护、社会保障体系的完善等财政和政策的支持力度。     

添加植酸酶日粮不同磷水平和钙磷比对生长肥育猪血液生化指标和骨骼性能的影响

本试验利用饲养试验方法,研究玉米-豆粕型日粮中添加植酸酶时不同磷水平和钙磷比对生长肥育猪血清指标和骨骼性能的影响。试验选用162头20kg左右的杜长大三元杂交猪进行2×3(磷水平×钙磷比)因子饲养试验,试验分2个阶段进行,前期为生长期体重在20-60kg,后期为肥育期体重为为60-100kg。结果表明:添加植酸酶日粮磷水平对血清磷含量和碱性磷酸酶的活性影响显著(P<0.05);钙磷比对血清钙和血清磷含量的影响显著(P<0.05);添加植酸酶日粮磷水平和钙磷比对生长肥育猪骨骼性能的影响不显著(P>0.05)。在添加植酸酶条件下,降低日粮磷水平不影响猪的正常生长发育。     

温度与盐度对老年低额蚤和蚤状蚤生长及生殖的影响_

温度与盐度对老年低额蚤和蚤状蚤生长及生殖的影响王丹丽，李明云，杨天勇，胡章（宁波大学，３１５０１０）关键词老年低额　，蚤状　，温度，盐度，生长，生殖ＥＦＦＥＣＴＳＯＦＴＥＭＰＥＲＡＴＵＲＥＡＮＤＳＡＬＩＮＩＴＹＯＮＧＲＯＷＴＨＡＮＤＲＥＰＲＯＤＵＣＴ...     

奶牛酮病发病机理研究进展

结合国内外目前研究状况较为详细的综述了奶牛酮病的病因与发病机理,并分析了奶牛酮病发生的生化机制。旨在对临床实践中很好的认识诊断和群体监控奶牛酮病提供较为系统的理论依据,保证奶牛群的高产和稳产,从而创造良好的经济效益。     

川渝地区鸡皮刺螨的流行与综合防治

鸡皮刺螨是鸡的一种体外寄生的节肢动物,流行十分广泛,对养殖业发展有严重影响。为了解鸡皮刺螨的发病情况,对川渝两地的126个鸡场进行流行病学调查,感染率高达71%。同时,介绍了鸡皮刺螨的生活史、危害及防治措施,是防制鸡皮刺螨不可多得的资料。     

Composition and diversity of fish and fish catches in closures and open-access fisheries of Kenya

Abstract  Catch composition, relative abundance and diversity of fish catches in open access and three old fisheries closures were compared and contrasted with previous ecological studies. There was less variation in catch community composition among the fishing grounds than the closures, suggesting that fishing has homogenised catch composition. The trap survey found that some parrotfish [ Leptoscarus vaigiensis (Quoy &Gaimard), Calotomus carolinus (Valenciennes) and Scarus ghobban Forsskål] were relatively more common and that some important predators of macro-invertebrates [ Balistapus undulatus (Mungo Park) and Cheilinus chlorourus (Bloch)] were less common in the fishing grounds than closures. Unexpectedly, and in contrast to visual census results, cumulative number of species in catch surveys was higher in open access than closures sites. This may result from fishers covering more area and habitat or a reduction in the catch of competitively subordinate and rare species by aggressive, early-caught fish that can dominate bait. Comparisons of ecological visual census surveys and fisheries-dependent methods indicated that small differences in catch composition can reflect larger ecological differences and that baiting methods can underestimate biodiversity. Ecological impacts of fishing and large-scale changes in marine ecosystems must be considerable given the many fisheries-dependent assessments report modest changes.