用地高辛标记的PCR－ELISA技术快速检测转基因鱼

取含有小鼠重金属螯合蛋白（mMT)基因启动子及人生长激素（hGH)基因的鱼样品，以Qiagene核酸纯化技术对鱼组织DNA进行纯化。常规PCR检测显示，mMT启动基因和hGH基因的PCR产物分别为240bp和130bp，与设计相符。PCR产物的核酸测序结果证实其扩增产物具特异性。地高辛-PCR（Dig-PCR)标记反应显示，2个基因均产生1条Dig标记的特异产物带。Dig-PCR-ELISA检测敏感性试验显示，对mMT启动子和hGH基因的检测敏感性可达10^-3，与常规PCR结合琼脂糖胶电泳检测方法相比，检测敏感性可提高至100-1000倍。试验证明，针对mMT和hGH基因所建立的Dig-PCR-ELISA技术特异可靠。     

Particle clearance and selection in three species of juvenile scallops

Juvenile scallops (<2 mm shell height) of three species (Placopecten magellanicus, Patinopecten yessoensis, Argopecten irradians) were fed mixed, unialgal cultures. Scallops were fed a total of six algal clones simultaneously and clearance rates were monitored using flow cytometric techniques. In another experiment, scallops were presented with natural assemblages of particulate matter as a food source. Data are presented on differences in clearance rates for the individual algal species as well as size-related differences of algal clones, and uptake of chlorophyll vs. non-chlorophyll cells, both within and between scallop species. Significant differences in clearance rates of individual algal species have been found within and between scallop species. Particle selection does not appear to be based upon size alone and is apparently based on other characteristics of the algae as well. The results demonstrate pre-ingestive sorting.     

Evaluation of the nutritive value of Leucaena leucocephala leaf meal, inoculated with fish intestinal bacteria Bacillus subtilis and Bacillus circulans in formulated diets for rohu, Labeo rohita (Hamilton) fingerlings

Nine isonitrogenous (35% crude protein approximately) and isocaloric (18.37 kJ g^?1) experimental diets (RLL20–BCFL40) were formulated with either raw or treated (inoculated with fish intestinal bacteria) Leucaena leucocephala leaf meal at 20%, 30% and 40% levels replacing other ingredients partially from a fish meal based reference diet (RD). Two specific strains of fish intestinal bacteria, Bacillus subtilis (isolated from Cyprinus carpio) and B. circulans (isolated from Oreochromis mossambicus) having extracellular cellulolytic and amylolytic activities, were used to inoculate Leucaena leaf meal for 15 days at 37°C. The crude fibre, cellulose and hemicellulose contents and the antinutritional factors, tannin, phytic acid and mimosine in the leaf meal decreased due to inoculation. However, free amino acids and fatty acids increased in the treated leaf meal. The response of rohu, Labeo rohita, fingerlings fed the experimental diets for 80 days was compared with fish fed a RD. Both the inclusion level and type of Leucaena leaf meal in diets significantly affected the growth performance of rohu. Fish fed diets containing inoculated Leucaena leaf meal performed better in comparison with those with the RD. On the basis of growth response, feed conversion ratio, protein efficiency ratio and apparent net protein utilization, diet formulated with 30%Leucaena leaf meal inoculated with B. circulans resulted in the best performance of rohu fingerlings followed by diet with 40%B. subtilis inoculated Leucaena leaf meal. The apparent protein digestibility (APD) was better in fish fed diets containing B. circulans inoculated leaf meal. An increasing level of raw Leucaena leaf meal was associated with a decrease in the carcass protein content of rohu fingerlings. The activity of α‐amylase increased with the increasing level of treated leaf meal in diets. Cellulase activity increased with increasing level of inclusion of raw leaf meal, and was comparatively lower in fish fed diets with treated leaf meal. Activities of protease and lipase were higher in fish fed the RD. The results showed that it is possible to incorporate Leucaena leaf meal inoculated with enzyme‐producing fish intestinal bacteria in carp diets up to 40% level of inclusion.     

Comparison of dot blot and PCR diagnostic techniques for detection of white spot syndrome virus in different tissues of Penaeus monodon

Comparison of PCR and dot blot diagnostic techniques for detection of white spot syndrome virus (WSSV) was made on different tissues of infected Penaeus monodon including eye stalk, eye stalk with eye, gills, cuticle, pleopod, periopods, uropods and telson. Dot blots of crude DNA extracted from infected tissue samples showed positive reactions with all the samples; however, the sensitivity of the dot blot was reduced with the purification of DNA samples extracted from pleopod, telson and uropod. PCR was found to be more sensitive when compared to dot blot. Both crude DNA and purified DNA samples extracted from all the tissues except for eye stalk with eye showed single step nested PCR positive reaction. The amplification of all or either of the three bands of 941 bp, 525 bp and 204 bp size varied with the tissues analysed. The severity of infection assessed by PCR amplification was found to be maximum in cuticle and telson followed by gill. Other tissues such as eye stalk, pleopod, periopods and uropod were observed to have mild infection. The maximum intensity of the PCR product was for the smallest amplified product of 204 bp followed by 525 bp and the weakest intensity was observed for the 941 bp size. The limitation of PCR due to inhibiting factors present in tissues could be overcome with the use of dot blot which gave positive reaction from the DNA extracted from eye stalk including the eye but yielded no amplification by PCR.     

Identification of Mycobacterium spp. isolated from snakehead, Channa striata (Fowler), and Siamese fighting fish, Betta splendens (Regan), using polymerase chain reaction–reverse cross blot hybridization (PCR–RCBH)

A variety of methods have been used to identify Mycobacterium spp. isolated from snakehead and Siamese fighting fish, including biochemistry, mycolic acid profiles and antibody-based methods. However, these methods are unable to differentiate between different species of Mycobacterium . Polymerase chain reaction (PCR) followed by reverse cross blot hybridization (RCBH) was adapted in this study to speciate aquatic mycobacteria. The method was highly specific for Mycobacterium spp. and identified the bacteria to species level with a detection limit of 100 fg DNA, equivalent to 20 mycobacteria. Twenty-nine isolates previously collected and cultured from Siamese fighting fish (10 isolates) and snakehead (19 isolates) during outbreaks of mycobacteriosis were analysed using PCR–RCBH. Six of the Siamese fighting fish isolates and nine of the snakehead isolates were identified as Mycobacterium fortuitum , while the remainder were classified as M. marinum . Notably, two isolates recovered from snakehead and Siamese fighting fish, previously identified as M. poriferae and M. piscicida , respectively, were confirmed to be M. fortuitum .     

荧光实时定量PCR技术及其在水产养殖研究中的应用

荧光实时定量PCR技术是近年来快速发展起来的一门新技术,它是核酸探针技术、荧光共振能量传递技术(Fluorescence Resonance Energy Transfer,FRET)和PCR技术的有机结合。与常规PCR相比,它具有特异性更强、有效解决PCR污染问题、自动化程度高、能较准确定量等特点。本文综述了荧光实时定量PCR技术的基本原理及几种广泛应用的荧光化学方法,并概述了荧光实时定量PCR技术在水产养殖研究领域的应用现状,并对荧光实时定量PCR技术的应用前景进行了展望。     

基于VB.NET和SQL Server肉牛场信息管理系统的研发

为了克服肉牛品种选育和饲养管理中存在操作不规范、数据记录不全、数据保存不完整等问题,提高肉牛养殖场信息化管理水平,提高企业的经济效益.本文根据肉牛饲养和牛场管理的需要,以Visual Studi0 2008为系统开发平台,利用VB.NET面向对象编程语言和SQL Server 2005大型关系数据库及Crystal Reports（水晶报表）技术进行系统开发.构建了基于VB.NET和SQL Server的肉牛场信息管理系统,系统包括文件、牛群管理、生产管理、育种管理、库存管理、人事管理、统计分析、系统维护和帮助九个主要功能模块.该系统实现了肉牛生产的系统化和牛场管理的信息化,促进肉牛品种选育的进程,降低企业的管理成本,有效地提高了肉牛养殖的效率.     

华亭县肉牛养殖业发展现状及对策建议

华亭县自古以来就是黄牛的主产区之一,又是秦川牛、早胜牛的边缘产区,养牛已成了当地农户发展致富的主导产业。本文在调查研究的基础上,对华亭县肉牛养殖业发展中存在的问题进行了综合分析,基础母牛保护力度不大、资金短缺、技术匮乏、基础设施落后,产业加工链短是制约华亭县肉牛养殖业发展壮大的主要瓶颈,针对以上问题,提出了一些可行的对策建议,以促进该县进一步加快肉牛改良、扩繁和规模化发展,降低养殖户养殖成本,提高经济效益。     

临泽县肉牛产业现状的调查与思考

肉牛产业是临泽县的主导产业。近年来,我县立足区位优势和资源优势,以建设全省牛羊产业大县和实施"张掖百万头肉牛基地工程"为契机,着力做大做强奶肉牛产业。通过政策扶持、科技助推、典型带动等措施大力推进标准化生产,实现农民增收。本文对临泽县肉牛发展的有利因素、存在问题进行分析,提出了肉牛发展对策和建议。     

大通养牛业发展现状及思路

近年来,大通县按照县委、县政府制定的实施意见、发展思路和规划,把畜牧业作为"牧业增效、农民增收"为奋斗目标。调整畜牧业内部结构,优化区域布局,逐步建立和发展奶牛、肉牛养殖生产基地。进行专业化生产,规模化经营,科学化饲养,转变传统养殖观念,逐步树立现代科学养殖观念,大力发展奶牛、肉牛等一批优势主导产业,逐步建立奶牛、肉牛养殖生产基地,推动全县畜牧业的快速发展。本文对大通县养牛业发展现状进行了详细分析,并对今后的发展思路进行了探讨。