牦牛源正呼肠孤病毒2型的检测和分离鉴定

旨在调查川西北牦牛哺乳动物正呼肠孤病毒（MRV）的感染情况并分离病毒。采用RT-PCR方法，对采自川西北15个牧场的72份牦牛腹泻粪便样本和其中5个牧场的15份腹泻牦牛血清样本进行MRV检测，阳性样本进一步用分型PCR确定其血清型。结果显示，粪便样本中MRV检出率为20.83%（15/72），血清2型的比例为60%（9/15）；血清样本中MRV检出率为40%（6/15），血清2型的比例为83.33%（5/6）；未检测到其他血清型。成功地从腹泻粪便中分离到1株MRV血清2型毒株（TCID₅₀为4×10^-8.56·mL^-1），并获得长度为23 587 bp的分离株全基因组，该分离株与中国猪源毒株的遗传关系最近；与GenBank中所有的MRV S1基因相比，该分离株有4个独特的氨基酸突变。本研究从牦牛中检测到MRV，并分离到1株牛源MRV血清2型毒株，为进一步研究MRV血清2型生物学特性奠定了基础。     

玫烟色棒束孢耐热菌株的筛选及对麦长管蚜的致病力测定

采用48℃恒温水浴热胁迫法对玫烟色棒束孢7个菌株进行了耐热性测定。结果表明,热胁迫45 min后,不同菌株的残存活孢率差异显著,其中IF-502、IF-904和IF-9606菌株耐热性较强,残存活孢率均达50%以上;IF-7606菌株耐热性最差,残存活孢率仅为28.70%;IF-14、IF-511和IF-609菌株耐热性居中,残存活孢率34.72%～36.00%。对耐热性强的IF-502、IF-904和IF-9606三个菌株对麦长管蚜若蚜的致病力测定表明, IF-904菌株在1×10~7孢子/mL时对麦长管蚜的致病力最高,校正死亡率达88.20%,致死中时最短,为2.67 d。     

油茶内生拮抗细菌的筛选、鉴定及防效

为筛选出对油茶炭疽菌有拮抗效果的生防菌株，以油茶健康叶为对象，平板对峙法筛选，利用形态和生理生化特征以及16S rDNA基因和gyrA基因序列对菌株进行鉴定，测定其抑菌活性，并将该菌株与枯草芽胞杆菌Y13复配进行林间防效试验。结果表明，对油茶炭疽病菌拮抗作用效果较强的内生菌株为HBMC—B05；菌落为乳白色，不透明，边缘不整齐，中间有凸起；分子鉴定结果显示菌株HBMC—B05为贝莱斯芽胞杆菌Bacillus velezensis，其对果生炭疽菌Collectotrichum fructicola，抑菌率可达到81.31%，对另外3种油茶炭疽病病原菌抑菌率可达到70%以上，对其他4种致病菌也有抑制作用；复合菌剂对油茶病害的田间防治效果达到59%以上。该菌株的抑菌具有广谱性，可为油茶病害的生物防治扩充菌种资源，具有广泛的应用前景。     

Ecological and landscape effects on genetic distance in an assemblage of headwater fishes

Environmental divergence along hierarchically structured longitudinal gradients may constitute barriers to gene flow in river networks for headwater specialised species. While known, this phenomenon has not been well studied, especially with regard to degree of headwater specialisation. We examined six headwater species that differ in habitat specialisation to assess whether patterns of differentiation vary according to geographic or environmental distance. We also identified regional environmental or anthropogenically induced fragmentation effects by comparing within‐drainage patterns of genetic distance across replicate watersheds. We used a comparative modelling framework to determine whether isolation by distance or isolation by resistance of large river habitats was a better predictor of genetic distance across species. The influence of reservoir presence and regional network characteristics that may influence the hydrology and size of large river habitats were also assessed. Resistance effects from large rivers were closely related to headwater specialisation, with increased specialisation leading to increased resistance and loss of drainagewide population connectivity. These results affirm that dendritic networks naturally fragment headwater specialised species. Further isolation from anthropogenic fragmentation was detected in two of the six drainages, indicating interactions with system‐specific conditions. Landscape variables related to the hydrology of large rivers also affected genetic distance in predicted ways, supporting the importance of large rivers in genetically structuring headwater species in drainage networks.     

Differences in the effectiveness of reproduction among ten breeding strains of the common carp Cyprinus carpio L. after stimulation of ovulation with carp pituitary homogenate or [(D‐Ala6, Pro9NEt) mGnRH‐a + metoclopramide] (Ovopel)

This study focused on the reproduction effectiveness of 10 breeding strains of common carp (Polish strains 2, 3, 6; Hungarian strains 0, W, 7; Lithuanian strain B; French strain F; Israeli strain D and Yugoslavian strain J) after stimulation of ovulation with carp pituitary homogenate or Ovopel. The percentage of females with recorded ovulation became higher after Ovopel treatment in as many as eight breeding strains (2, 3, 6, 0, W, B, F, D). The interaction between the spawning agent and the breeding strain was statistically significant (P ≤ 0.05) for the mass of spawn and non‐significant for traits determining the quality of eggs. Among the 10 breeding strains, in 4 (3, 7, B, J) the least‐squares means (LSM) for the weight of egg (g) were higher for fish stimulated with Ovopel. Among the strains tested, in seven (3, W, F, J, 2, 7, B), the LSM for the percentage of living embryos after 36 h incubation were higher for fish treated with Ovopel. The evaluation of reproduction effects based on the values of such parameters as the percentage of ovulating females, weight of eggs (g) and the percentage of live embryos (36 h) showed that after Ovopel treatment the poorer effects were obtained only for strains 6, 0 and D.     

不同发酵条件对荔枝产香菌发酵茶品质的影响

经过长时间的工艺优化,国内不同地区的黑茶风味已趋于稳定,为了进一步改善发酵茶品质,生产更具特色的发酵茶,利用课题组前期已筛选出的一株适用于茶叶发酵的优良产香菌ZYF3单菌发酵青砖茶原料(一芽四五叶)。在实验室条件下,以灭菌后青砖茶为发酵原料,探究该菌株在不同接菌量、不同茶坯含水量、不同发酵温度和不同发酵时间下对发酵茶感官品质和理化品质的影响。结果表明:在不同发酵条件下,产香菌ZFY3发酵茶品质差异显著,当接种量为1.2%～1.3%(mL/g)、茶坯含水率为45%、发酵温度为32℃、发酵时间为20 d时,发酵茶综合品质最佳。不同发酵条件能明显改善荔枝产香菌发酵茶品质,并且促进发酵茶特殊风味的形成。     

安徽省猪伪狂犬病毒的分离鉴定及其主要毒力基因分子特征

猪伪狂犬病是由猪伪狂犬病毒(porcine pseudorabies virus,PRV)引起的一种急性、热性传染病。自2011年底以来,PRV发生了变异,传统的PRV弱毒疫苗已不能对PRV变异株提供完全保护,这给我国PR防控带来了巨大挑战。为了解安徽省PRV流行特征及其主要毒力基因遗传变异情况。利用PCR技术、细胞接种试验、电子显微镜观察、间接免疫荧光试验及兔体接种试验等方法,对安徽省临诊病例中疑似PRV感染的病猪进行病原检测及PRV分离鉴定,并通过设计6对特异性引物对PRV分离株主要毒力基因(gE、gI、TK、gB、gC、gD)进行克隆及测序分析。2016—2018年安徽省临诊病例中共分离鉴定15株PRV;PRV分离株主要毒力基因序列均与2011年后国内PRV变异株同源性较高;与2011年前国内PRV经典株序列比对,PRV分离株gE、gB、gC及gD基因存在多个位点的一致性替换、插入或缺失,且gE、gC基因多位点突变位于其重要的抗原表位区。本研究分离的15株PRV均为变异株,变异株已成为安徽省主要的流行毒株。15株PRV分离株的gI、TK基因序列较为保守,而gE、gC蛋白抗原表位区域氨基酸的突变可能导致其毒力及抗原性发生改变。部分PRV分离株与邻近地区PRV序列同源性均为100%,可能与频繁跨省调运生猪、跨区引种等原因有关。     

金刺梨黑斑病病原菌的分离及鉴定

金刺梨黑斑病主要于开花期危害幼嫩的果梗,果梗受害后产生黑色干腐病斑,导致整个花序从果梗处脱落。对黑斑病病原菌进行了分离和鉴定,通过形态学特性、致病性测定、rDNA-ITS序列分析,初步确定该病主要由拟茎点霉属(Phomopsis Sacc.)的Phomopsis.capsici真菌引起。     

2株毒死蜱降解菌的分离鉴定及其混合降解特性研究

从生产毒死蜱农药厂采集的活性污泥中分离筛选得到2株降解效率较高的毒死蜱降解菌,命名为D1、D3,根据表型特征、生理生化特性和16 S rRNA基因序列相似性分析,将其鉴定为苍白杆菌属(Ochrobactrum sp.)和副球菌属(Paracoccus sp.)细菌。2株菌以最佳配比(1∶1.25)混合施用时,与单菌相比,毒死蜱降解效率可提高12%~26%;以混合菌株为研究对象,发现其对毒死蜱最适降解温度为30℃,最适降解pH值为7.0,最适NaCl浓度为0.5%;混合菌株施入土壤后,可保持较高的定殖能力和降解效率。     

Protective efficacy of an H5/H7 trivalent inactivated vaccine produced from Re-11, Re-12, and H7-Re2 strains against challenge with different H5 and H7 viruses in chickens

We developed an H5/H7 trivalent inactivated vaccine by using Re-11, Re-12, and H7-Re2 vaccine seed viruses, which were generated by reverse genetics and derived their HA genes from A/duck/Guizhou/S4184/2017(H5 N6)(DK/GZ/S4184/17)(a clade 2.3.4.4 d virus), A/chicken/Liaoning/SD007/2017(H5 N1)(CK/LN/SD007/17)(a clade 2.3.2.1 d virus), and A/chicken/Guangxi/SD098/2017(H7 N9)(CK/GX/SD098/17), respectively. The protective efficacy of this novel vaccine and that of the recently used H5/H7 bivalent inactivated vaccine against different H5 and H7 N9 viruses was evaluated in chickens. We found that the H5/H7 bivalent vaccine provided solid protection against the H7 N9 virus CK/GX/SD098/17, but only 50–60% protection against different H5 viruses. In contrast, the novel H5/H7 trivalent vaccine provided complete protection against the H5 and H7 viruses tested. Our study underscores the importance of timely updating of vaccines for avian influenza control.