原位杂交技术及其在水产养殖动物病毒性疾病诊断中的应用

本文介绍了原位杂交技术的发展概况、基本原理和应用价值,并概述了原位杂交技术在水产养殖动物(包括鱼、虾、贝类)病毒性疾病的检测、定位和体内表达模式等方面的应用。此外,还对该技术的应用前景进行了展望。     

云南干热河谷旱坡地南洋樱植物篱水土保持效益研究

对云南元谋干热河谷旱坡地水土流失和土壤退化严重的地区,引进豆科速生树种南洋樱,以30 cm×40 cm株行距、品字形双行等高栽植快速形成植物围篱。结果表明,南洋樱植物篱在元谋干热河谷旱坡地生长良好,并且在种植3年间,治理区径流发生次数明显减少,年径流模数减少了29.13×104m3/km2,减少率76.90%;土壤年退化减少12 986.8 t/km2,减少率76.02%。研究区降水量与径流量间者的相关系数由建植篱前的显著相关变为不显著;有机质和钾流失减少量高达3 111.64 kg/hm2和3276.57 kg/hm2。南洋樱植物篱在控制旱坡地水土流失、减少土壤侵蚀和改善土壤养分等效益显著。     

黄山市生态旅游存在的问题与可持续发展对策

分析了黄山市生态旅游的资源优势,探讨了生态旅游资源开发中所存在的问题,从法制建设、科学规划、人才建设等方面提出了相关的建议及对策。     

Genomic differentiation with isolation by distance along a latitudinal gradient in the spotted-leg treefrog Polypedates megacephalus

The patterns of isolation by distance (IBD) entailing increased genetic differentiation among populations have aroused extensive concerns for evolutionary biologists. Although the IBD may act on spatial processes contributing to the genetic differentiation among populations in anuran species, the factors shaping the IBD of frogs among populations in natural systems are largely unknown. Here, we studied the genetic differentiation among six populations with 24 individuals of the spotted-leg treefrog along a latitudinal gradient (1860.31 km) based on 1020 single nucleotide polymorphisms from restriction site-associated DNA sequencing. The results showed that the genetic diversity differed significantly among populations and that the insular populations had higher genetic diversity than the mainland populations. Furthermore, we also found a significant genetic differentiation among populations (F_ST = 0.277) and no sign of inbreeding (F_UNI = −0.145). The IBD was detected for all populations, and a higher degree of the IBD was indicated when controlling for the effects of the isolation between Hainan and mainland populations caused by the Qiongzhou Strait. Our findings suggest that the form of the Qiongzhou Strait plays a key role in shaping the genetic diversity and population differentiation in treefrogs.     

牛支原体间接免疫酶标组织化学检测方法的建立

为建立检测牛支原体的间接免疫组织化学(简称免疫组化)方法,分别以鸡抗牛支原体(Mycoplasma bovis)IgY、羊抗鸡IgG-HRP为一抗和二抗,对牛支原体菌体涂片和牛支原体阳性肺脏组织切片进行免疫组化染色,并对染色条件进行优化,确定组织触片及组织切片的最佳免疫组化条件。在最佳条件下对病死犊牛病料切片进行免疫组化检测,并以细菌分离培养和PCR方法进行验证。结果显示,免疫组化法阳性标本与分离培养和PCR结果相符,而阴性对照和替代试验均为阴性。结果表明本试验建立的间接免疫组化方法可用于检测临床病例组织样本及培养物中的牛支原体,具有特异性和可靠性,为探究该病原在机体内的定位、动态分布规律及致病机理提供了手段。     

猪附红细胞体和弓形体病继发猪瘟的诊断与防制

2004年2月至3月，湖北剂州某市八宝、中洲、老城、胡家岗和荆闸等乡镇自然村多家个体养猪专业户在断奶仔猪中爆发一种以体温高、贫血、黄疸、精神沉郁，先便秘后拉稀为特征的传染病。医治不见好转，先后死亡450多头。于3月中旬和下旬分2批将病猪送到我院进行诊断，经剖检、实验室检验和动物试验，最后确诊为猪附红细胞体和弓形体病继发猪瘟，采用超量免疫及综合防制措施，使本病得到了有效的控制。     

脾酪氨酸激酶参与酿酒酵母甘露聚糖诱导绵羊瘤胃上皮细胞β-防御素-1(SBD-1)的表达

为了探索脾酪氨酸激酶(Syk)是否参与酿酒酵母甘露聚糖(S.c M)诱导绵羊瘤胃上皮细胞(ORECs)β-防御素-1(SBD-1)表达的过程,首先利用免疫组化、RT-PCR和免疫荧光等方法检测Syk在ORECs内的表达情况;然后采用qPCR和Western blot方法检测S.c M刺激ORECs后Syk的表达变化,同时用Western blot方法检测Syk的磷酸化水平;接着用3条Syk特异性siRNAs(#1、#2和#3)转染ORECs 24 h后,qPCR检测Syk mRNA的表达变化,筛选出干扰效果最佳的Syk siRNA;最后用效果最佳的siRNA和Syk特异性抑制剂R406分别处理ORECs后,采用qPCR和ELISA检测SBD-1的表达变化,以确定Syk在S.c M诱导SBD-1表达过程中的作用。结果显示:Syk在ORECs内表达;且S.c M刺激ORECs后Syk的mRNA和蛋白表达水平显著高于未刺激组(P<0.01或P<0.05),S.c M刺激ORECs不同时间(5、15、30、45和60 min)均能使Syk发生磷酸化,且刺激15 min后磷酸化水平达到最大(P<0.01);此外,Syk的3条特异性siRNAs转染ORECs后Syk的表达均降低,且Syk siRNA#2的抑制效果最明显(P<0.01);同时Syk siRNA#2和R406均能极显著降低S.c M诱导ORECs SBD-1的表达(P<0.01)。上述结果表明,Syk参与S.c M诱导ORECs SBD-1的表达。     

Characterization of a new gene (SLC10) with a spliced leader from Taenia solium

An unknown gene, SLC10, was cloned by spliced leader-based polymerase chain reaction from Taenia solium. The full length of SLC10 was found to be 635 bp, encoding an 18.223 kDa protein. ELISA results showed that none of 70 normal and 75 cysticercosis sera samples reacted with purified recombinant SLC10 protein. Using an immunohistochemical method, it was revealed that the native SLC10 protein distributed extensively in inner cyst walls but not in the scolex in Cysticercus cellulosae. Together with predicted results, it is suggested that the SLC10 protein is a non-secretory structural protein, which is not involved in induction of the host's immune reactions against infection at least at the larval stage.     

俄罗斯引进鲁梅克斯新品种在黑龙江省的推广种植

黑龙江省北部全年低温时间较长,西部地区土地盐碱化严重,是制约黑龙江农牧业发展的两大主要因素。为了筛选出与黑龙江省气候和土壤类型相适应的牧草,本研究对从俄罗斯引进的鲁梅克斯新品种‘ЩавельЧемпион’在黑龙江省不同积温带的5个具有代表性的种植点阿城、肇州、逊克、嫩江和呼玛的农艺性状和营养价值进行研究。结果表明,该品种在肇州的生长状况明显差于其它地区,而在嫩江的生长状况明显优于其它地区;产量则表现为阿城、逊克和嫩江较高,产量均在150t·hm-2以上,肇州和呼玛略低,但产量也均在120t·hm-2以上。从营养数据可以看出,阿城种植区的粗蛋白及粗脂肪含量显著高于其它地区(P0.05),其中粗蛋白为18.41%,粗脂肪为3.82%;肇州种植区的粗纤维含量显著低于其它地区,含量为10.75%,而粗灰分含量显著高于(P0.05)其它各种植区,达到14.85%;嫩江种植区钙含量显著高于其它地区;肇州种植区的干物质含量显著高于其它地区。结果表明,引进新品种‘ЩавельЧемпион’适应性强,能够在黑龙江省不同区域不同土壤上种植并推广。     

Ensiling characteristics of total mixed ration prepared with local feed resources in Mozambique and their effects on nutrition value and milk production in Jersey dairy cattle

To effectively utilize total mixed ration (TMR) prepared with locally available feed resources, we studied the nutritional value and milk production in Jersey dairy cattle fed a local general diet (LGD) and fermented TMR in Mozambique. Ten head of Jersey dairy cattle with 337 ± 19.8 kg body weight, aged 3–4 years in mid location were used in this study. The LGD diet was designed following the general feeding method of local smallholding farms; it contained native grass, Napier grass, wheat bran, and mineral–vitamin mix. Fermented TMR was prepared using Napier grass, corn bran, wheat bran, formula feed, and mineral–vitamin mix. Fermented TMR was preserved as good quality, with a relatively low pH and high lactic acid content. Compared to LGD, fermented TMR significantly improved the dry matter intake and digestibility in dairy cattle. Milk yield was significantly higher in dairy cattle fed with TMR than in those fed with LGD, by 3.75 L/d; milk quality was not significantly different between treatments. The results confirmed that LGD had a low dry matter intake and milk yield, and fermented TMR prepared with local feed resources can attain good quality and improve milk yield in dairy cattle in Mozambique.