庆阳市采用塑料大棚发酵畜禽粪污模式初探

为了提高粪污发酵速度、缩短发酵时间,减少环境对粪污的承载和消纳能力,笔者通过采用塑料大棚模式发酵粪污,以提高粪污内部温度,添加一定的微生物菌剂,采用高温好氧堆积发酵,在西北气候条件下,使小型养殖场(合作社)粪污发酵比常温堆积发酵时间缩短了40～100 d,使粪污尽快得到还田利用,减少了环境对粪污的承载力。     

13种蒲公英属植物核型似近系数聚类分析

为探究蒲公英属植物亲缘关系,本研究对13种蒲公英属（Taraxacum. F. H. Wigg.）植物进行了核型似近系数聚类分析,并计算了物种间遗传距离。结果表明：当核型似近系数接近0.6960时,13种蒲公英可以聚为4大类群;当核型似近系数接近0.4641时,13种蒲公英共属一类。异苞蒲公英与狭戟片蒲公英的亲缘关系最近,其似近系数最大,为0.9969,核型进化距离最小,为0.0031;多裂蒲公英和亚洲蒲公英的亲缘关系最远,似近系数为0.3343,核型进化距离为1.0959。本研究结果为蒲公英属植物的亲缘关系辨析、种质鉴定及物种进化分析提供依据。     

猪细小病毒NS基因原核表达与多克隆抗体制备

旨在制备抗猪细小病毒(PPV)非结构蛋白共有氨基酸的NS多肽多克隆抗体。根据GenBank(MK993540)公布的PPV基因组序列,克隆其非结构蛋白NS1、NS2共有基因序列(NS基因),并进行生物信息学分析。进而将NS基因克隆至原核表达载体pET-32a,构建重组质粒pET-32a-NS,转化至大肠埃希菌Rosetta(DE3)中进行诱导表达,利用镍柱亲和层析技术纯化表达的重组多肽,用重组多肽免疫Balb/c小鼠制备多克隆抗体。结果显示,PPV杨凌株NS基因长258bp,编码86个氨基酸的多肽,是具有亲水性的非跨膜NS多肽,具有大量的B细胞线性表位。NS多肽在37℃、0.8mol/L IPTG条件下,诱导6h有大量的可溶性表达。免疫印迹试验结果显示,该多肽具有较好的抗原性。用纯化NS多肽免疫小鼠后获得鼠抗NS多肽的血清抗体效价为1∶12800。用制备的NS多克隆抗体检测纯化的NS重组多肽及真核表达的NS1蛋白,均能检测出相应特异性条带,为进一步研究NS蛋白在PPV致病过程中的作用奠定了基础。     

裸果木种群遗传多样性特点及与地理气候因子关联研究

本文利用ISSR标记技术,选取15对引物,对分布于西北荒漠区12个裸果木种群的遗传多样性以及与地理气候因子的相关性进行了分析。结果表明：群体内多态性位点百分率（PPL）为88.81%,种群水平为69.93%;Nei’s基因多样性指数（He）变化范围为0.2356~0.2697,Shannom信息指数（I）变化范围为0.3509~0.4057,具较高的遗传多样性水平。基因分化系数（Gst）为0.2790,种群间遗传分化较小,种群内存有丰富的遗传变异。经Mantel检验,种群遗传距离与地理距离有相关性（R²=0.2144）。利用NTSYSpc-2.1软件进行UPGMA法聚类,各种群样品间的遗传相似系数在0.81～0.99之间,按照遗传距离远近,裸果木12个种群可分为4大类群。经相关性分析,种群遗传多样性与纬度、海拔存在正相关（P<0.05）与年均温存在负相关（P<0.05）。     

羊粪浸提液对燕麦种子萌发特性的影响

为研究羊粪对燕麦（Avena sativa）种子萌发特性的影响,以不同浓度羊粪浸提液（0、25%、50%、75%、100%）处理的7个品种（‘燕王’、‘牧王’、‘爱沃’、‘枪手’、‘魅力’、‘领袖’和‘贝勒’）的燕麦种子进行萌发试验。结果表明：100%羊粪浸提液显著抑制枪手的发芽率（P<0.05）,其他6个品种燕麦种子发芽率没有受羊粪浸提液的影响;羊粪浸提液显著降低‘牧王’、‘魅力’、‘领袖’和‘贝勒’的发芽势（P<0.05）,且随着浸提液浓度的升高,种子发芽势逐渐降低;随着浸提液浓度的升高,各品种燕麦种子的发芽指数均有逐渐降低的趋势;100%浓度浸提液显著降低种子活力指数（P<0.05）,而50%浸提液能显著提高‘燕王’、‘牧王’、‘领袖’和‘贝勒’的活力指数（P<0.05）。     

Effects of Mixed Caulis Lonicerae japonicae Extracts on Fracture Healing and Anti-inflammation

This experiment was aimed to study the effects of mixed Caulis Lonicerae japonicae extracts on fracture healing and inflammation. Rabbit right radius fracture model was induced by dental drill in order to study the medicine's effect on fracture healing. Moreover,the contents of Ca,P and alkaline phosphatase in serum were measured to explore the influences of mixed Caulis Lonicerae japonicae extracts on serum biochemical indexes of fractured rabbits.RAW264.7 cell was induced by LPS in order to research the effect of medicine on inflammation. The concentrations of TNF-α,IL-1β,IL-6 and NO were measured by ELISA. The results showed that the rabbit fracture model was successfully prepared by dental drill, and the extracts of Caulis Lonicerae japonicae could increase the levels of Ca and alkaline phosphatase and reduce the P content in the serum of fractured rabbits. The TNF-α,IL-1β and NO levels in RAW264.7 cell induced by LPS were significantly inhibited by the extracts with concentrations ranging from 50 to 200 μg/mL (P<0.05). However, the extracts had no significant effect on the level of IL-6 (P>0.05). These results demonstrated that the extracts of mixed Caulis Lonicerae japonicae could promote the fracture healing and possessed good resistance inflammation effect.     

Study on Expression of Bcl-2 and PCNA Genes in Testicular Tissue of Chickens,Quails and Chicken-quail Hybrids

In order to investigate the molecular mechanism of stunted growth testis of chicken (♂) and quail (♀) hybrids. 84 testicular tissue samples of New Roman cocks, male Korean quails and chicken-quail hybrids at different developmental stages were collected,the mRNA expression of Bcl-2 and PCNA genes in testicular tissue of cocks, quails and chicken-quail hybrids at different growth stages were detected using Real-time PCR. The results showed that the Bcl-2 and PCNA genes mRNA expression patterns in testes of chickens and quails at different growth stages were similar.The Bcl-2 gene was fluctuated in a whole, and decreased to the lowest value, then recovered rapidly and remained at a high level. The PCNA gene had a very significant peak value.Compared with the chicken and quail,the expression of Bcl-2 and PCNA genes mRNA in hybrids had no obvious change, which indicated that the pattern of the Bcl-2 gene regulation apoptosis in testis was different from chicken and quail at the hybrid testicular development process, and there was no obvious cell appreciation process.It was suggested that the molecular influencing factors of testicular dysplasia of chicken and quail hybrids were related to the abnormal expression of Bcl-2 and PCNA genes mRNA in testes.The results provided an important reference for the further study of the mechanism of testicular dysplasia between chicken and quail interspecific hybrids.     

Isolation,Identification and Genetic Evolution Analysis of Mink Serratia marcescens

In order to determine drug resistance, pathogenicity and genetic of the pathogenic bacteria in mink of Heilongjiang province, three strains were isolated from dead minks and identified. Based on colony morphology, microscopic characteristics, biochemical test and 16S rRNA sequencing. The antibiotic susceptibility to 24 antibiotics were detected by using standard K-B paper method, and virulence assay, phylogenetic analysis were carried out at the same time. The result showed that the isolates were highly tolerance to antibacterials, could cause 100% death of mice in experimental group and were closely related to strains from activated sludge and volcanic deposits. It showed that Serratia marcescens infection of mink existed in Heilongjiang province. Strains resistance phenomenon was serious and had a strong pathogenicity, and might be originated from the environment. It was the first time that the pathogenic Serratia marcescens isolated from the mink was reported. The results provided information for the detection, identification, diagnosis, treatment and prevention of related diseases.     

Isolation,Identification and Phylogenetic Analysis of Important Functional Genes of Porcine Pseudorabies Virus LC Strain

To find out the reason of the reproductive failure in pregnant sows in a hoggery in Guangdong, the mixture with brain, lymph nodes and lungs of farm abortion stillbirth were identified by PCR, virus isolation and culture, tissue culture infective dose (TCID₅₀) assay, homology and phylogenetic analysis of the important functional genes (gB, gC, gD and gE) and animal test. The results showed that the mixture was proved to be porcine pseudorabies virus (PRV) positive samples. The typical cytopathogenic effect was induced in the third passage of Vero cell and the titer of the fifth passage was 10^-6.8/0.1 mL. The sequence analysis and phylogenetic relationship of gB, gC, gD and gE genes showed that it was a Chinese PRV variant, which was named as LC strain. The typical pseudorabies clinical and pathological symptoms were presented in 12-week-old piglets inoculated with LC strain. The results demonstrated that a local pseudorabies virus had been isolated, suggesting that the Bartha-K61 vaccine was not fully effective for controlling the current epidemic of pseudorabies in China.     

产箱环境对母貂母性行为和仔貂生长发育的影响

试验探讨了产箱环境对母貂母性行为和仔貂存活、生长、发育的影响。4种新型产箱类型有A、AB、AC、AD,B型为传统产箱。通过母貂的做窝行为和对5日龄仔貂的取回试验判断产箱环境对母貂母性行为的影响,通过出生重、出生个数、仔貂死亡率、平均体重、体表温度来判断产箱环境对仔貂存活、生长的影响。结果表明,仔貂的出生重和出生个数不受产箱环境的影响;AC型产箱能够增强母貂的母性,提高1~14日龄仔貂的存活率和平均体重。新型产箱在保温性能和产仔保活方面都优于传统产箱。