抗禽流感病毒H5和H9亚型血凝素特异性单克隆抗体的研制及应用

以禽流感题H5和H9亚型病毒分别免疫Balh／c小鼠，取其脾细胞与SP2／0的骨髓瘤细胞融合，用血凝抑制试验(HI）检测细胞培养上清，结果获得了6株特异性单克隆抗体，其中抗禽流感题亚型病毒血凝素特异性单克隆抗体细胞株3株，分别命名为4B6、4A3、3H1；抗H9亚型病毒血凝素单克隆抗体细胞株3株，命名为6E6、6B6和5B4。这些单克隆抗体小鼠腹水HI效价为2^13-15，细胞培养上清抗体HI效价为2^7-8。研究结果表明，所有这些单克隆抗体仅与试验的相应题或ID亚型病毒株发生特异性反应，而不能与鸡新城疫病毒、鹅新城疫病毒、鹅腺病毒和鸡产蛋下降综禽征病毒(EDS76)等反应。实验室检测结果证明，应用这些单克隆抗体能在24h内迅速检测出相应的禽流感病毒。所有这些单克隆抗体将在禽流感的预警预报工作中发挥重要作用。     

Cryptic species and population genetic structure of Plasmopara viticola in São Paulo State,Brazil

Downy mildew (Plasmopara viticola) is one of the most important diseases in grape-growing areas worldwide, including Brazil. To examine pathogen population biology and structure, P. viticola was sampled during the 2015/16 growing season from 516 lesions on nine grape cultivars in 11 locations in subtropical areas of São Paulo State, Brazil. For identification of cryptic species, a subsample of 130 isolates was subjected to cleaved amplified polymorphic sequence (CAPS) analysis, and for 91 of these isolates the ITS1 region was sequenced. These analyses suggest that the population of P. viticola in São Paulo State consists of a single cryptic species, P. viticola clade aestivalis. Seven microsatellite markers were used to determine the genetic structure of all 516 P. viticola isolates, identifying 23 alleles and 55 multilocus genotypes (MLGs). Among these MLGs, 34.5% were clonal and represented 93% of the isolates sampled. Four dominant genotypes were present in at least five different locations, corresponding to 65.7% of the isolates sampled. Genotypic diversity (Ĝ = 0.21–0.89) and clonal fraction (0.58–0.96) varied among locations (populations). Most populations showed significant deviation from Hardy–Weinberg expectations; in addition, excess of heterozygosity was verified for many loci. However, principal coordinate analysis revealed no clusters among locations and no significant isolation by distance was found, suggesting high levels of migration. The results indicate that downy mildew epidemics result from multiple clonal infections caused by a few genotypes of P. viticola, and reproduction of P. viticola in São Paulo State is predominantly asexual.     

2014年-2016年广西地区猪繁殖与呼吸综合征病毒Nsp2和ORF5基因的遗传多样性分析

为了解广西地区猪繁殖与呼吸综合征病毒(PRRSV)流行毒株的遗传进化情况,对2014年-2016年来自广西各地的部分PRRSV阳性病料进行Nsp2和ORF5基因的扩增和测序分析.结果获得34个Nsp2基因序列和45个ORF5基因序列,均属于美洲型毒株.Nsp2基因间核苷酸序列的同源性为91.8％~100％,与PRRSV美洲型毒株VR-2332、CH-1a、JXA1及NADC30株核苷酸序列的同源性分别为81.3％~84.3％、88.9％~92.1％、94.3％~99.3％和73.5％~75.1％,而与PRRSV欧洲型毒株LV株核苷酸序列的同源性为51.5％~53.2％.ORF5基因间核苷酸序列的同源性为82.8％~100％,与PRRSV美洲型毒株VR-2332、CH-1a、JXA1及NADC30株核苷酸序列的同源性分别为83.7％~99.5％、85％~95％、83.8％~99.7％和83.2％~86.4％,而与PRRSV欧洲型毒株LV株核苷酸序列的同源性为62.4％~64.5％.基于Nsp2和ORF5基因推导的氨基酸序列绘制的遗传进化树中,广西地区的毒株主要分布在以JXA1为代表的Ⅳ亚群.表明当前广西PRRSV流行毒株以JXA1株为代表的高致病性美洲型毒株为主,各毒株Nsp2和ORF5基因序列存在一定的差异,尚未发现欧洲型毒株和美洲型NADC30类毒株.     

Genetic diversity of livestock-associated MRSA isolates obtained from piglets from farrowing until slaughter age on four farrow-to-finish farms

During a previous longitudinal study, performed on four farrow-to-finish farms (A to D), samples were taken from twelve sows, their offspring, and the environment on various occasions over six months to study the MRSA presence. During the present study, a selection of the obtained MRSA isolates were typed by multiple-locus variable-number tandem-repeat analysis (MLVA), Pulsed Field Gel Electrophoresis (PFGE), spa typing, and SCCmec typing to study the genetic diversity of LA-MRSA isolates and to determine possible MRSA sources for pig(let)s. PFGE, spa typing, and SCCmec typing revealed the presence of one or few dominant genotype(s) per farm. In contrast, 212 MLVA types were detected on the four farms, forming one cluster on farm A, three on farm B, four on farm C and two on farm D. The genotype, found on farm A was unique for this farm. Farms B, C and D shared one cluster. In general, MLVA types from these clusters were isolated from piglets, sows, and the environment on various sampling events. Piglets carried MLVA types both related and unrelated to their mother sows’ MLVA types at farrowing and onwards. In conclusion, molecular typing revealed that within a farm one or a few dominant strain(s) are widespread. Potential MRSA sources for piglets were mother sows, the environment and other piglets.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0089-4) contains supplementary material, which is available to authorized users.     

马鹿的起源进化与遗传多样性研究进展

马鹿是非常重要的物种资源,由于栖息地的流失和人为干扰进行近亲繁殖等导致野生马鹿数量急剧减少,而家养马鹿多经过改良,因此马鹿的纯种数量锐减。对马鹿进行分子遗传学研究不仅可以加深人们对马鹿起源和物种形成的认识,还能帮助开展遗传多样性保护研究。随着高通量测序技术、分子生物学和生物信息学的迅速发展,马鹿的起源进化研究已发展到全基因组水平,并取得了一定的成果。马鹿的起源进化研究从最初对体态外貌和染色体核型的研究逐渐发展到对DNA序列与生理指标的研究。文章回顾了近年来国内外对马鹿起源进化和遗传多样性方面的研究,从起源时间、起源地和迁徙路线等方面揭示了马鹿的演化历史,介绍了父系、母系和常染色体研究方面分析了马鹿遗传多样性选取的不同分子标记,为进一步揭示马鹿种群的遗传变异、分化情况、迁徙路线和系统发育关系等提供基础信息,同时为马鹿遗传资源的利用和保护以及马鹿产业的良性发展提供重要的借鉴与参考。     

H5亚型禽流感DNA疫苗质粒pCAGGoptiHA5对高致病力禽流感病毒攻击的免疫保护

对H5亚型禽流感DNA疫苗质粒pCAGGoptiHA5的免疫效果进行了研究。pCAGGoptiHA5分别以100和10μg剂量一次或两次免疫3周龄SPF鸡，首次免疫后4周以同样剂量和途径进行第二次免疫，一次免疫后4周、两次免疫后2周分别用100LD50的HPAIV A／Goose／GuangDong／1／96（H5N1）鼻腔途径进行攻击，观察发病与死亡情况，分别于攻毒后第3、5、7天采集喉头及泄殖腔拭子进行病毒分离、滴定检测排毒情况，同时检测免疫后、攻毒前及攻毒后血清HI抗体、NT抗体以及AGP抗体的动态变化。结果，100μg pCAGGoptiHA5一次免疫、100μg pCAGGoptiHA5两次免疫以及10μg pCAGGoptiHA5两次免疫均可对免疫鸡形成100％完全保护（不发病、不致死、不排毒），10μg剂量pCAGGoptiHA5一次免疫可对免疫鸡形成100％的保护（不发病、不致死），结果表明，pCAGGoptiHA5作为疫苗效果良好、成本低廉，有望成为预防H5亚型高致病力禽流感的高效、安全新型基因工程疫苗。     

毛乌素沙地南缘生态过渡带植被物种多样性的研究

对毛乌素沙地南缘生态过渡带流动半流动沙丘、固定半固定沙地、盐化丘间低地和缓坡丘陵梁地4种不同生境的植被物种组成及多样性进行研究。结果表明:随着沙化程度的加重,从缓坡丘陵梁地,经盐化丘间低地、固定半固定沙地到流动半流动沙丘,多年生草本植物逐渐减少,一年生草本植物及沙生灌木、半灌木逐渐增加;物种丰富度指数、多样性指数及均匀度指数均下降。     

Development and characterization of monoclonal antibodies to subgroup A avian leukosis virus

Avian leukosis virus subgroup A (ALV‐A) is a retrovirus which infects egg‐type chickens and is the main pathogen of lymphoid leukosis (LL) and myeloid leukosis (ML). In order to greatly enhance the diagnosis and treatment of clinical avian leukemia, two monoclonal antibodies (MAbs) to ALV‐A were developed by fusion between SP2/0 and spleen cells from mice immunized with expressed ALV‐A env‐gp85 protein. Using immunofluorescence assay (IFA), two MAbs reacted with ALV‐A, but not with subgroups B and J of ALV. Western blot tests showed that molecular weight of ALV‐A envelope glycoprotein recognized by MAbs was about 53 kD. Isotyping test revealed that two MAbs (A5C1 and A4C8) were IgG1 isotypes. These MAbs can be used for diagnosis and epidemiology of ALV‐A.     

荒漠草原自然恢复中植物群落组成及物种多样性

摘要：通过调查封育3、4、5、6、7年及未封育荒漠草原群落的特征,研究了不同封育年限下草原植物群落组成及生物多样性的变化,以期探明封育对荒漠草原群落恢复的影响。结果表明,封育改变了草原植物群落组分,表现为随封育年限延长,多年生草本植物,特别是短花针茅(Stipa breviliora)等一些地带性物种比例增加;封育草原的Shannon Wiener指数、Pielou指数、Simpson指数均比未封育草地高,封育5年草原的多样性指数最大。     

云南水牛mtDNA D-loop遗传多样性研究

[目的] 探究云南3个水牛品种(德宏水牛、滇东南水牛、盐津水牛)的mtDNA D-loop区的遗传多样性与母系起源。[方法]采用PCR扩增、测序及生物信息学方法。[结果] 本研究共分析了215条云南水牛mtDNA D-loop全序列,检测到107个多态位点,定义了86种单倍型。结果表明,云南3个水牛品种的mtDNA遗传多样性非常丰富,其中,滇东南水牛的遗传多样性最高(Hd: 0.946±0.017,Pi: 0.0162±0.0018),盐津水牛的遗传多样性最低(Hd: 0.805±0.063,Pi: 0.0141±0.0031)。系统发育分析结果表明,在3个云南水牛品种中检测到2个主要支系(A和B支系及其亚支系)及一个稀有支系C,其中A支系为主要支系(79.07%),且经历了强烈的群体扩张。[结论] 云南3个水牛品种具有丰富的mtDNA遗传多样性,主要有A与B两个母系起源。