口蹄疫重组鸡痘病毒活载体疫苗在豚鼠、猪体内的毒性、分布及抗体消长规律

利用临床观察、病理解剖、PCR、RT-PCR、ELISA、中和抗体检测等方法,对口蹄疫(FMD)重组鸡痘病毒(FPV)在豚鼠、仔猪体内的毒性、分布以及抗体消长规律进行研究。结果表明,FMD重组鸡痘病毒免疫的动物在整个试验期间,未表现出明显的临床症状和不良反应;病理组织切片检测无明显的组织学变化;PCR、RT-PCR检测证明,豚鼠、猪免疫FMD重组鸡痘病毒后,在心脏、肝脏、脾脏、肺脏、肾脏、肌肉、脑、肠系膜淋巴结内检测到FPVDNA和FMDV DNA,且在大部分组织能存在3 d左右;FMD重组鸡痘病毒均可诱导免疫动物产生较高水平的抗FMDV特异性抗体和中和抗体,验证了所构建重组FPV的生物安全性及良好的免疫原性,为其他哺乳动物实验提供了必要的基础数据。     

稀释液中添加谷氨酰胺对绒山羊冷冻精液品质的影响

在绒山羊冷冻精液稀释液中分别添加0mmol/L、2.5mmol/L、5.0mmol/L、7.5mmol/L和10.0mmol/L五个梯度的谷氨酰胺,以确定冷冻效果最好的谷氨酰胺添加浓度,优化绒山羊冷冻稀释液配方。结果表明:添加5.0mmol/L谷氨酰胺组精子的活率、顶体完整率和生存指数均极显著或显著地高于其他试验组(P0.01,P0.05),而畸形率极显著或显著低于其他试验组(P0.01,P0.05);在脂质过氧化反应和抗氧化酶活性方面,添加5.0 mmol/L组MDA浓度极显著低于其他试验组(P0.01),而CAT活性极显著高于其他试验组(P0.01)。总之,在稀释液中添加5.0mmol/L谷氨酰胺,在冷冻—解冻过程中能起到很好的保护精子的作用。     

谷子RNA干扰相关酶类基因家族的鉴定与分析

Dicers酶类、Argonautes蛋白以及RNA依赖的RNA聚合酶(RNA-dependent RNA polymerases,RDRs)是RNA干扰(RNA interference,RNAi)机制中重要的核心蛋白,但在谷子(Setaria italica)中尚无系统报道.为了研究谷子中与RNA干扰相关酶类基因的特征,本研究对谷子的RNA干扰相关酶类基因家族进行了蛋白质理化性质、亚细胞定位预测、蛋白质保守基序、基因保守结构域、基因家族成员间系统发育关系以及组织特异性表达谱分析.研究共发现24个与谷子RNA干扰相关酶类基因,包括7个DCL(Dicers),13个AGO(Argonautes),4个RDRs.系统进化树分析表明,这些家族被分为3个进化支.同一家族基因成员具有共同的保守结构域.虽然大多数基因可同时在不同发育时期的叶、茎和穗中表达,但其在各时期的穗和茎中的表达量最高.本研究为详细探讨这些基因在谷子生殖和生长发育中的表观遗传修饰作用提供了理论依据.     

Phylogenetic analysis of S1 gene of infectious bronchitis virus isolates from China

Between 2006 and 2009, seven strains of infectious bronchitis (IB) virus (IBV) were isolated from vaccinated chicken flocks on different chicken farms in China. The pathogenic characters of seven IBV strains were assessed. Each of the seven strains was infective to the test chickens and could induce an immune response. The results from chicken embryo cross-neutralization assays showed that these strains were antigenically distinct from classic IBV strains of H120, M41, Conn, and Gray. Compared to H120 vaccine strain, point mutation, short insertion, and deletion occurred at many positions in the S1 protein of the seven strains. Five of the seven strains had the motif (HRRRR), which was identical to that of the epidemic IBV strains in China. Two new motifs (HRLRR and RRIRR) emerged in the isolated strains. The homology of the nucleotide and amino acid sequences of the S1 gene among the seven isolates was 81.7%-99.7% and 79.0%-99.4%, respectively. These seven strains were also genetically different from the vaccine strains and non-China IBV strains but closely related to large numbers of Chinese strains. The seven isolates and 36 reference IBV strains were clustered into six distinct groups (I-VI). The seven strains were categorized into groups I, II, and III, forming a big phylogenetic branch, which is closely related to Chinese IBVs, whereas the vaccine strains belonging to group VI are genetically distant from groups I, II, and III. The results from this study indicate that different IBV strains cocirculate in the chicken population in China.     

猪流行性腹泻病毒山西分离株S基因遗传变异分析

为分析山西地区猪流行性腹泻病毒（PEDV）的遗传变异情况,试验利用RT-PCR方法对2014-2015年山西省疑似猪流行性腹泻的阳性病料进行克隆和测序,获得4个S基因片段,并对其基因序列和推导的氨基酸序列与国内外毒株进行比对分析。序列分析结果显示,4株PEDV山西分离株的S基因与CV777 vaccine相比,在170~171 bp之间插入12个核苷酸,在401~402、454~455 bp之间均插入3个核苷酸,在461~468 bp之间缺失6个核苷酸。4株PEDV山西分离株S基因之间核苷酸和氨基酸同源性分别为99.2%~99.8%和98.6%~99.7%,与2011-2015年中国流行毒株、CV777 vaccine、attenuated DR13、CV777的核苷酸同源性分别95.0%~98.5%、93.2%~93.6%、93.2%~93.7%、93.7%~94.4%,氨基酸同源性分别为96.2%~98.9%、91.9%~92.6%、92.1%~92.9%、92.9%~94.0%。遗传进化树分析结果表明,PEDV S基因分为3个群,4株PEDV山西分离株属于第一群,与2010年以后国内流行毒株（除AH-M、SQ2014）的亲缘关系较近,与2010年以前中国流行毒株、2个日本株、7个韩国株、2个疫苗株的亲缘关系较远。研究结果提示山西省流行的PEDV发生较明显的变异,需研发新的疫苗来控制PEDV的暴发。     

紫茎泽兰浸提液对牧草种子发芽和幼苗生长的影响

紫茎泽兰被我国列为16种有害外侵物种之首,在西南地区大规模侵入草场、农田、森林,很有必要了解其化感效应,研发无害化处理与资源化利用相结合的技术。试验以广泛分布于四川省凉山州的白三叶、黑麦草和紫花苜蓿为材料,比较了新鲜(extract of fresh E. adenophorum, EFA)和腐熟紫茎泽兰的浸提液(extract of composed E. adenophorum, ECA)对牧草种子萌发和幼苗生长的影响,以明确这种外侵植物对草场植被的危害作用和腐熟处理效果。随EFA浸种和培养浓度的提高,不同程度地抑制牧草种子发芽和幼苗生长。当浓度达到100 mg/L EFA时,牧草根毛消失,根尖向上卷曲,离开EFA,根尖发黑,甚至死亡。相反,用铜绿假单胞菌(Pseudomonas putita sp.)和高温纤维菌(Clostridium thermocellum sp.)组成的混合菌剂腐熟紫茎泽兰后,ECA提高牧草种子发芽率、发芽指数和活力指数,并促进幼苗生长,其最大增幅达到21.11%(种子发芽率),24.12%(苗高)和22.48%(生物量)。此外,用100 mg/L EFA浸种和培养牧草幼苗,抑制胚乳中的淀粉、蛋白质和肌醇磷酸盐水解,显著降低游离氨基酸、可溶性糖、可溶性磷,以及幼苗中的叶绿素、根系活力及硝酸还原酶活性,ECA则相反。这可能是EFA抑制(或ECA促进)牧草种子发芽和幼苗生长的重要生理原因之一。因此,EFA含有对牧草有害化感的物质,抑制其种子发芽和幼苗生长;腐熟紫茎泽兰可降解毒素,刺激牧草种子发芽,促进幼苗生长,实现紫茎泽兰的无害化处理与资源化利用,为当地农业和畜牧业生产提供大量的优质有机肥源。     

云南省六个水稻产区稻瘟病菌三个无毒基因的组成及其致病型

为明确水稻抗性基因Piz-t、Pib和Pii的有效性,利用无毒基因AvrPiz-t、AvrPib和Avr-Pii的特异性引物对自云南省6个水稻产区采集并分离获得的348株稻瘟病菌Magnaporthe oryzae菌株进行PCR扩增检测,并测定其对仅含Piz-t、Pib和Pii基因的水稻抗性单基因系IRBLzt-T、IRBLb-B和IRBLi-F5品种的致病性,明确这3个无毒基因在云南省水稻产区组成及分布。结果表明,在348株稻瘟病菌菌株中,分别有51.7%、46.8%和15.8%的菌株含有无毒基因AvrPiz-t、AvrPib和Avr-Pii,GT8、GT2、GT5、GT6、GT1、GT3、GT4和GT7基因型菌株检测频率分别为24.7%、21.8%、21.0%、16.7%、4.9%、4.0%、3.4%和3.4%;分别有4.9%、29.2%、41.1%和24.7%的菌株含有3、2、1和0个无毒基因;云南省稻瘟病菌群体总多样性指数水平较高,为2.81,其中滇中水稻产区的最高,为2.97;在348株稻瘟病菌菌株中,分别有89.1%、63.2%和38.5%的菌株对单基因系IRBLzt-T、...     

No effect of exogenous melatonin on development of cryopreserved metaphase II oocytes in mouse

Background

This study was conducted to investigate effect of exogenous melatonin on the development of mouse mature oocytes after cryopreservation.

Results

First, mouse metaphase II (MII) oocytes were vitrified in the open-pulled straws (OPS). After warming, they were cultured for 1 h in M₂ medium containing melatonin at different concentrations (0, 10⁻⁹, 10⁻⁷, 10⁻⁵, 10⁻³ mol/L). Then the oocytes were used to detect reactive oxygen species (ROS) and glutathione (GSH) levels (fluorescence microscopy), and the developmental potential after parthenogenetic activation. The experimental results showed that the ROS level and cleavage rate in 10⁻³ mol/L melatonin group was significantly lower than that in melatonin-free group (control). The GSH levels and blastocyst rates in all melatonin-treated groups were similar to that in control. Based on the above results, we detected the expression of gene Hsp90aa1, Hsf1, Hspa1b, Nrf2 and Bcl-x1 with qRT-PCR in oocytes treated with 10⁻⁷, or 10⁻³ mol/L melatonin and untreated control. After warming and culture for 1 h, the oocytes showed higher Hsp90aa1 expression in 10⁻⁷ mol/L melatonin-treated group than in the control (P < 0.05); the Hsf1, Hsp90aa1 and Bcl-x1 expression were significantly decreased in 10⁻³ mol/L melatonin-treated group when compared to the control. Based on the above results and previous research, we detected the development of vitrified-warmed oocytes treated with either 10⁻⁷ or 0 mol/L melatonin by in vitro fertilization. No difference was observed between them.

Conclusions

Our results indicate that the supplementation of melatonin (10⁻⁹ to 10⁻³ mol/L) in culture medium and incubation for 1 h did not improve the subsequent developmental potential of vitrified-warmed mouse MII oocytes, even if there were alteration in gene expression.     

入境肉种鸡检出衣原体阳性及其处理

目的随着加入世界贸易组织,我国海关检疫压力增大,为了维护国家主权和国际声誉,加强出入境检验检疫势在必行。衣原体病是一种重要的人畜共患病,也给养禽业带来巨大的经济损失,必须给予足够的重视,鹦鹉热衣原体对多种动物宿主有广泛的寄生性引起不同动物的多症候群疾病,是世界动物卫生组织动物名录疾病。方法对入境的一批肉种鸡进行抽样检验,按照中华人民共和国出入境检验检疫行业标准用间接血凝检测鹦鹉热衣原体病,做出试验报告。结果经检测出现阳性,肉鸡携带有衣原体病毒。结论根据中华人民共和国进出境动植物检疫法第二章进境检疫第十六条检出二类传染病、寄生虫病的动物,退回或者扑杀,同群其他动物在隔离场或者其他指定地点隔离观察。     

机器人足球比赛防守与进攻队形转换的研究

随着机器人足球的发展,机器人防守和进攻的队形对于比赛显得尤为重要。为了加强机器人足球比赛中的防守和进攻,文章通过分析足球在不同区域时防守队形的选择、防守与进攻的队形转换、以及在极端情况下剩余机器人的有效利用等几个问题,研究和设计机器人防守和进攻队形转换的算法和策略。机器人队形的选择采用人性化的方法,大大提高了队形的形成速度,在极端状态下也做出相应的策略来加强进攻。通过实验表明,该策略增加了防守和进攻的效率,提高了进球的几率。