产气荚膜梭菌B型C58-1株β1毒素基因的克隆表达

利用PCR技术,从B型产气荚膜梭菌中国标准株C58-1株扩增出β1毒素基因,连接pMD18-T 载体筛选阳性克隆,然后用限制性核酸内切酶BamHⅠ和SalⅠ对其进行酶切,回收927 bp的β1毒素基因片段,将其定向克隆到载体pET-32a中,获得重组质粒pETβ927。将pETβ927转化至受体菌BL21(DE3)中,其表达产物经His-Trap FF预装柱纯化、SDS-PAGE 检测目的蛋白大小和分布及Western blotting检测其反应原性。结果表明,完整的β1毒素基因大小为1 011 bp,与GenBank发表的B型和C型产气荚膜梭菌β1毒素蛋白序列同源性达99.4%以上;SDS-PAGE结果显示重组目的蛋白在大肠杆菌中成功表达,融合蛋白大小为 54 ku,在超声波裂解上清和包涵体中均有分布,但以包涵体为主。Western blotting检测结果显示表达的重组蛋白可与特异性血清抗体发生免疫反应,表明β1毒素蛋白具有较好的反应原性。     

扶正解毒散及其超微粉体外抗鸡传染性法氏囊病病毒药效比较

采用鸡胚法和细胞法2种体外抗病毒试验比较扶正解毒超微粉(FZSM)和扶正解毒散(FZ)的抗鸡传染性法氏囊病病毒(IBDV)效果。鸡胚法:在FZSM和FZ 2种煎煮液对鸡胚安全浓度范围内分别选取3个浓度,采用先加煎煮液后接种病毒和先接种病毒后加煎煮液2种方式,接种10日龄SPF鸡胚,测定2种煎煮液对IBDV ELD50的影响。细胞法:在FZSM和FZ 2种煎煮液对鸡胚成纤维细胞(CEF)安全浓度范围内分别选取3个浓度,采用先加煎煮液后接种病毒和先接种病毒后加煎煮液2种方式,加入到长成单层的CEF中继续培养,观察并统计各孔的细胞病变(CPE)。结果显示,FZSM和FZ 2种煎煮液均可以降低IBDV的病毒滴度和CPE抑制率,高浓度效果优于低浓度;同浓度FZSM效果优于FZ;预防作用效果优于治疗作用效果。结果表明FZSM和FZ均具有体外抗IBDV的作用,且FZSM效果优于FZ。     

不同紫花苜蓿品种在河西走廊盐碱地的适应性研究

通过田间试验测定不同紫花苜蓿品种的株高、分枝数、茎叶比、生物量、出苗率和越冬率,评价16个紫花苜蓿品种在河西走廊盐碱地的适应性,遴选出适宜河西走廊盐碱地种植的紫花苜蓿品种。结果表明:参试的16个紫花苜蓿品种,骑士T、SR4030和巨能的出苗率较高,甘农3号的出苗率最低;16个紫花苜蓿品种间株高差异不显著(P0.05);BR4010、WL343HQ、阿迪娜、亮苜2号和WL363HQ的分枝数显著大于标靶和金皇后的分枝数;岩石和德宝的叶茎比显著高于金皇后和骑士3的叶茎比;德宝、亮苜2号和骑士T的产量大于其他品种;亮苜2号和骑士3的越冬率显著高于甘农3号和SR4030的越冬率;阿迪娜和WL343HQ的倒伏率最小,骑士T和标靶的倒伏率最大。以株高、分枝数、茎叶比、生物量、出苗率和越冬率为变量因子,通过聚类分析将16个紫花苜蓿品种聚为6类,综合评判结果认为,亮苜2号、骑士T、德宝适宜种植于河西走廊盐碱地。     

尕海湿地CH4、CO2和N2O通量特征初步研究

2011年7月-2012年7月,采用静态箱-气相色谱法同步研究了尕海4种典型湿地类型的CH₄、CO₂和N₂O通量及其与温度因子的关系,并估算了其全球变暖潜势值(GWP)。结果表明,尕海湿地的CH₄、CO₂和N₂O通量具有明显的空间变化特征,CH₄、CO₂和N₂O通量最小值分别为亚高山草甸(-0.014±0.126) mg/(m²·h),沼泽湿地(137.17±284.51) mg/(m²·h)和高山湿地(-0.008±0.022) mg/(m²·h),而最大值分别为沼泽湿地(0.498±0.682) mg/(m²·h),高山湿地(497.81±473.09) mg/(m²·h)和草本泥炭地(0.094±0.117) mg/(m²·h);同时CH₄、CO₂通量有明显的时间变化特征,通量最大值分别出现在2011年的7-10月和2012年的5-7月,而后降低并维持相对稳定的变化趋势;5 cm地温、气温、地表温度及箱内温度与4种类型湿地CO₂通量呈极显著正相关关系(P<0.01),与高山湿地CH₄通量均存在显著正相关关系(P<0.05),与其他3种湿地类型CH₄通量的相关性均较差,但与4种湿地类型N₂O通量无显著相关性;尕海草本泥炭地、沼泽湿地、高山湿地和亚高山草甸4种类型湿地的温室效应贡献潜力依次为35.311,13.520,34.816和30.236 t CO₂/(hm² ·a), 沼泽湿地能够显著降低温室效应。     

Variants of the recently discovered avian gyrovirus 2 are detected in Southern Brazil and The Netherlands

A genome of a virus preliminarily named avian gyrovirus 2 (AGV2), a close relative to chicken anemia virus, was recently discovered in a chicken in the state of Rio Grande do Sul, Southern Brazil. To study the occurrence of AGV2 in Rio Grande do Sul and the neighboring state Santa Catarina, a number of adult chickens (n=108 and n=48, respectively) were tested for the presence of AGV2 DNA. An AGV2-specific PCR was developed, optimized and used to analyze DNA extracted from clinical samples. AGV2 DNA was detected in 98/108 (90.7%) of samples collected in the state of Rio Grande do Sul and 29/48 (60.4%) of the samples collected in the state of Santa Catarina. In order to check whether AGV2 DNA would be detected in samples from a geographically distant region, DNA from brain samples of 21 diseased chickens from the Netherlands were tested independently, by the same method. In such specimens, 9/21 (42.9%) brain tissue samples were found to contain AVG2 DNA. Sequence analysis of some of the PCR products demonstrated that the amplified AGV2 sequences could vary up to 15.8% and could preliminarily be divided in three groups. This indicated the occurrence of variants of AGV2, which may reflect differences in geographical origin and/or in biological properties. The data presented here provides evidence that AGV2 seems fairly distributed in chickens in Southern Brazil and that AGV2 also circulates in the Netherlands. Besides, circulating viruses display genetic variants whose significance should be further examined, particularly to determine whether AGV2 would play any role in chicken diseases.     

不同生理时期梅花鹿血液GSH-Px含量测定及其纯化

研究不同生理时期梅花鹿全血中GSH-Px含量变化及其纯化技术,为开发鹿血抗衰老资源奠定理论基础.用DINB法测定梅花鹿的溶血液、血细胞内容物、血浆和血细胞细胞膜中GSH-Px含量并计算全血GSH-Px含量;利用10％～100％饱和度的(NH4)2SO4盐析和柱层析技术纯化GSH-Px;用SDS-聚丙烯酰胺凝胶电泳测定其亚基相对分子质量.试验结果表明:全血GSH-Px含量以生茸期最高为(1 973.07士25.43)U·mL-1,与配种期的(1 727.74士12.46)U·mL-1差异极显著(P＜0.01),与生茸前期的(1 961.83士16.54)U·mL-1差异不显著(P＞0.05);GSH-Px的初始比活力为24.9 U·mg-1,经纯化后得到GSH-Px 0.37 mg· mL-1,最终比活力1 347.7U·mg-1,纯化倍数为54.1倍,回收率25.00％;鹿血GSH-Px亚基的相对分子质量为17.2 ku.结果提示生茸期鹿血的GSH-Px含量最高,开发利用价值最大,GSH-Px含量与生理特性相关;鹿血GSH-Px盐析条件为40％～80％饱和度的(NH4)2SO4,此时所得GSH-Px纯化倍数较高.     

CoCl_2对酸胁迫下多花黑麦草种子萌发及幼苗抗性的影响

本研究以多花黑麦草（Lolium multiflorum）为试验材料,采用不同浓度CoCl2溶液浸种的处理方法,就CoCl2对酸胁迫下多花黑麦草种子萌发、幼苗生长及抗逆性的影响进行了探讨。酸胁迫过程中多花黑麦草种子的发芽势、发芽率、发芽指数、活力指数、叶绿素含量和超氧化物歧化酶（SOD）活性随着处理液酸度的增强显著下降,根长抑制指数、丙二醛（MDA）含量、 O-2·产生速率和H2O2含量明显增大,过氧化物酶（POD）活性先增加后下降,细胞膜稳定指数（MSI）下降。0.1 mmol·L-1 CoCl2明显促进酸胁迫下多花黑麦草种子萌发,提高幼苗叶绿素含量、芽长、地上部干质量和SOD活性,降低MDA含量和POD活性。从整体上看,0.1 mmol·L-1 CoCl2对多花黑麦草种子萌发及幼苗生长有促进作用,并提高了多花黑麦草的抗酸胁迫能力。     

宁夏典型草原土壤微生物特征对不同管理方式的响应

草地土壤微生物是土壤养分循环的推动力,可灵敏反映草地生态环境变化,常被用来评价草地土壤质量。本研究以宁夏典型草原为对象,研究了封育、放牧和水平沟改良管理方式下土壤微生物特征及其与土壤有机质的相关性。结果表明,试验区0~40 cm土层土壤微生物数量表现为细菌>放线菌>真菌;细菌和放线菌数量随封育年限增加呈增加趋势,真菌以封育5年草地最高;微生物总数量和微生物量碳、氮含量均表现为随封育年限增加而增加,放牧草地最低;水平沟改良草地微生物特征未表现出明显的优越性;土壤微生物数量表现为5~15 cm土层最高,25~40 cm土层最低;土壤微生物数量与有机质含量之间存在极显著正相关关系。     

Serologic survey for Leptospira spp. in captive neotropical felids in Foz do Igua?u, Paraná, Brazil

Leptospirosis is a bacterial zoonosis of worldwide distribution and is endemic in tropical countries, where rodents and other wild mammals are abundant and may act as reservoirs. Leptospirosis has become a concern in captive wild animals, due mostly to their exposure to contaminated urine or environment. Although domestic cats (Felis catus) have been reported refractory to leptospirosis, serology and disease in captive wild felids is still unclear. In this study 57 adult, clinically healthy felids, including 1 Geoffroy's cat (Leopardus geoffroyi), 3 jaguarundis (Puma yagouaroundi), 17 margays (Leopardus wiedii), 22 little spotted cats (Leopardus tigrinus), and 14 ocelots (Leopardus pardalis) kept in captivity at the Sanctuary at the Itaipu Binacional hydroelectric power plant (Bela Vista Biological Sanctuary), Foz do Iguacu City, Paraná State, Brazil, were serologically surveyed for the presence of antibodies against 28 serovars of Leptospira spp. by microagglutination test (MAT). Two animals (3.5%) were seropositive: one male ocelot to the serovar Cynopteri (titer 100) and one female margay to Autumnalis (100) and Butembo (200). The captive-born, 5-yr-old ocelot had been solitary housed in an individual cage. The approximately 21-yr-old wild-caught margay was also kept individually. None of the tested animals showed signs ofleptospirosis. During a study conducted 4 yr previously in the same facility, this particular margay also tested positive for the same two serovars, among others. The present study indicates that the felids tested for Leptospira spp. by MAT were exposed to serovars, but did not demonstrate clinical signs of disease. Comparison with a previous study suggests that serovar titers may vary over time and that leptospirosis dynamics remains unclear in wild felids.