禽流感病毒H5HA、H7HA、H9HA亚型夹心ELISA方法的建立

将禽流感病毒(AIV)H5、H7、H9血凝素亚型标准血清中的IgG作为包被抗体,H5、H7、H9血凝素亚型单克隆抗体作为第二抗体,建立了双抗体夹心ELISA(AC-ELISA)方法,并优化了反应条件.结果表明:单抗和标准血清最适工作质量浓度分别为5、10 mg/L,标准阳性血清于4℃包被12 h,二抗作用条件为37℃作用1 h,封闭条件为37℃作用1 h,酶标抗体工作条件为1∶5 000稀释后,37℃作用1 h,底物作用条件为37℃作用10 min.阴阳性抗原临界值为0.274～0.332.本方法具有很高的特异性和敏感性.     

秋丰白玉异品系比较试验

近年来,蚕品种秋丰×白玉有不少改良品系,为摸清秋丰×白玉各品系之间的异同,根据省农业厅蚕种公司的部署,在德清县进行了秋丰×白玉各原种品系及其一代杂交种的比较试验,现将试验结果报告如下。     

Ontogeny of testicular inhibin and activin in ducks: An immunocytochemical analysis

The ontogeny of testicular inhibin/activin in ducks was investigated. Testicular localization of three inhibin/activin subunits (α, β_A and β_B) was determined in embryonic and newly hatched ducks from 12 days of incubation to 1 day of age, in immature ducks and in adult ducks. In the duck embryonic testis, positive α‐subunit immunostaining was first detected in the Leydig cells and Sertoli cells on day 15 of incubation, whereas β_A‐subunit and β_B‐subunit immunostaining were found in Sertoli cells and primary germ cells on day 18 of incubation. In 1 month old ducks, intense staining of α‐subunit was present in the seminiferous epithelium consistent with localization in Sertoli cells and primary germ cells, and the immunostaining of the β_A‐ and β_B‐subunit was also present in Sertoli cells and primary germ cells. Specific immunostaining with inhibin/activin α‐, β_A‐ and β_B‐subunits antisera occurred in Sertoli cells in the adult duck testes. In conclusion, it was shown that, in the duck testis, the majority of α‐, β_A‐ and β_B‐subunits are colocalized in Sertoli cells with a certain degree of staining in germ cells and the α‐subunit is present in Leydig cells of embryonic testes before day 18 of incubation. These results indicate that Sertoli cells and possibly germ cells in the embryonic testes of late stage of incubation and newly hatched ducks, immature ducks and mature ducks may produce bioactive inhibin dimers, inhibin A and inhibin B, as a possible regulator of follicle‐stimulating hormone secretion. Free inhibin/activin subunits and their dimers may also play an autocrine/paracrine role in the development of the testis and spermatogenesis. Furthermore, early onset of the α‐subunit in duck testes indicates that it may have an autocrine/paracrine effect on steroid hormones, which is important for sex differentiation.     

鸭禽流感免疫抗体血凝检测及其应用

鸭是禽流感传播链中的一个重要环节,能否有效防控鸭禽流感的关键之一是使用科学的检测方法.目前,是参照禽流感血凝(HA)和血凝抑制(HI)试验标准(GB/T18936-2003)[1],但是,鸭体内含有非特异性抑制因子,血清中的唾液酸残基会模拟红细胞受体,同红细胞受体竞争与流感病毒血凝素的结合,因而会导致非特异性抑制结果(假阳性)的出现,用鸡红细胞悬液检测水禽禽流感血清抗体时,常常存在非特异性凝集现象,出现前带和后带现象,有时能严重影响结果的判定.为此,本文采用不同种家禽的红细胞悬液做重复试验,探讨水禽血清的处理方法及不同种的家禽红细胞悬液对水禽禽流感血清抗体检测的影响, 在临床应用中,可为鸭禽流感免疫技术提供参考依据.     

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。     

泛素激活酶抑制剂对猪精子获能状态、膜重构及体外受精的影响

【目的】评估泛素激活酶(E1)对猪精子获能、膜重构和体外受精的影响。【方法】选取5头长白猪采集精液,以硫醇酯法评估泛素激活酶(ubiquitin activating enzyme, E1)抑制剂(TAK-243)对精子中泛素(ubiquitin, Ub)与E1结合的影响;将精子分为鲜精组(Fresh)、DMSO组(DMSO)、获能组(Capacitated)及2.4、4.8和7.2 nmol/L TAK-243组,鲜精组用2 mL PBS重悬精子沉淀;获能组用2 mL获能液重悬精子沉淀;DMSO组及2.4、4.8和7.2 nmol/L TAK-243组分别用2 mL含0.07%DMSO及2.4、4.8和7.2 nmol/L TAK-243的获能液重悬精子沉淀,使用微生物动(静)态图像检测系统检测精子的动力学参数;以Western blotting法检测精子的酪氨酸磷酸化水平;Zn²⁺荧光探针检测精子的Zn²⁺含量;花生凝集素染色检测精子的顶体膜重构;免疫荧光法检测顶体表面精子黏附蛋白(AQN-1、AWN)的表达;体外受精法检测TAK-243...     

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.     

猪伪狂犬病基因缺失疫苗的制备、安全性、免疫原性、保存期测定及区域试验

为了提供有效的伪狂犬病疫苗，用鸡胚成纤维细胞扩大培养了PrV HB-98突变株（TK^-/gG^-/LacZ^＋），研制了伪狂犬病基因缺失疫苗，并对该疫苗经肌肉接种、经口等免疫途径的最小免疫剂量进行了测定，同时也对4批疫苗的安全性、效力、免疫期和保存期进行了检测；同时将4批疫苗用于免疫23个猪场的母猪、新生仔猪和育肥猪进行区域试验。测定结果表明，疫苗经上述两种途径接种对不同阶段猪的最小免疫剂量均为10^5.0 TCID50；10倍免疫剂量的疫苗对初生仔猪、15日龄仔猪和妊娠母猪是安全的，免疫猪能抵抗强毒的攻击；疫苗在4℃和-20℃下分别可保存6个月和12个月。对伪狂犬病毒抗体阴性的70日龄商品猪和种猪的免疫期为6个月。田间试验表明，4批猪伪狂犬病基因缺失疫苗安全有效，并可用于仔猪发病时的紧急接种。为猪伪狂犬病基因工程疫苗的制备与应用提供了有力的依据。     

三江源地区高寒草地不同退化程度土壤特征研究

为了解区域尺度上不同退化程度高寒草地的土壤特征,在对三江源地区进行多样点采样和室内分析的基础上,研究草地退化对高寒草地不同土层的土壤物理和化学等特征的影响。结果表明:随着退化程度加剧,不仅草地群落组成出现明显变化,地上/地下生物量显著下降,而且土壤各理化性质也受到不同程度的影响。土壤容重随草地退化程度的加剧和土壤深度的增加而增大;土壤有机碳随草地退化程度的加剧而显著下降,尤其是0～10cm土层;草地退化加剧导致土壤总氮和硝态氮含量呈逐渐下降趋势,且对0～10cm土层影响更为显著;土壤铵态氮含量在草地轻度退化阶段最高,且受草地退化影响不显著。相关性分析表明,不同土层的土壤有机碳和总氮与生物量之间均呈显著正相关,多元逐步回归分析也显示土壤有机碳和总氮含量与绝大部分草地演替阶段的生物量显著相关,表明土壤有机碳和总氮可作为衡量三江源地区草地退化程度的重要土壤性状指标。人工草地的建植可明显增加地上/地下生物量,但短期内对土壤的改良效果并不显著,极度退化草地的土壤恢复将是一个缓慢的过程。