密码子优化的甲型H1N1流感病毒HA基因DNA疫苗在小鼠体内免疫保护效力研究

为了提高甲型H1N1流感病毒HA基因DNA疫苗的免疫原性,本研究将流感病毒rPan09(HA和NA来自A/California/04/09,其余6个片段来自PR8的重组病毒)的HA基因通过人工合成的方法将其密码子优化为哺乳动物体内偏嗜性密码子opti-HA,同未优化的rPan09的HA基因分别与真核表达载体pCAGGS连接构成重组质粒pCA-optiHA和pCA-HA转染293T细胞,48 h后采用间接免疫荧光的方法检测HA基因的体外表达情况,结果显示重组质粒pCA-optiHA的蛋白表达量明显高于pCA-HA。为评价这两种重组质粒的免疫及保护效力,选取6~8周龄雌性BALB/c小鼠,将质粒pCA-HA、pCA-optiHA以100μg/只的剂量,进行后腿肌肉多点注射,同时设立空载体pCAGGS对照,共免疫3次,每次间隔2周。结果表明DNA疫苗pCA-optiHA可显著提高小鼠的体液免疫和细胞免疫应答水平。三免2周后用107.45EID50的rPan09采用滴鼻方式进行攻毒,用Real-time PCR及制作肺组织石蜡切片检测DNA疫苗的保护效力。结果表明,pCA-optiHA免疫组的保护效力明显高于pCA-HA免疫组。本研究为进一步研究和设计有效的甲型流感病毒DNA疫苗奠定了基础。     

伊犁绢蒿荒漠草地围封后的土壤动态

本研究对伊犁绢蒿(Seriphidium transiliense)荒漠草地的土壤理化性质及光谱进行测定,探讨围封对土壤理化性质的影响和草地光谱的响应。结果表明,土壤含水量在各土层随植物生长期呈递减趋势,5月与9月土壤含水量随围封年限的延长总体上呈增加趋势,并对群落光谱和土壤含水量间关系进行建模,R2=0.63;土壤有机质随封育年限增加呈升-降-升的趋势,围栏内外有机质差值在5月随封育年限的增加而增加,封育7年、10年草地有机质差值,植物生长期内呈线性递减。通过室内土壤光谱及野外裸地光谱对土壤有机质进行建模,均取得较好效果,R2分别为0.64与0.38。研究表明,草地土壤含水量及有机质对于草地恢复状况较为敏感,可用于该类草地封育状况的监测。     

日粮补充苹果渣或有机矿对鹅生产性能、蛋品质及繁殖性能的影响

本研究旨在评估日粮单独添加苹果渣或补充苹果渣和有机微量元素对鹅生产、繁殖性能及蛋品质的影响.试验将产蛋期的360只鹅随机分为3组,每组120只(30只/重复),对照组饲喂基础日粮,处理组分别饲喂苹果渣日粮和苹果渣+有机微量元素日粮,试验持续14周.结果:苹果渣补充或不补充有机矿较对照组显著提高了孵化率和仔鹅存活率(P<...     

The effect of genotype,choice feeding,and season on organically reared broilers fed diets devoid of synthetic methionine

In response to the impending ban on synthetic methionine in organic poultry diets, researchers have focused on finding alternative strategies to supply this amino acid. The objectives of this study were to assess performance and carcass characteristics of broilers fed diets devoid of synthetic methionine by using 1) a slow-growing and a fast-growing genotype, 2) choice-feeding management (supplying grain and a complementary premix in 2 separate feeders), and 3) pasture access and seasonal variation. Inclusion of fish meal and high percentages of soybean meal enabled the specific genotype methionine requirement to be met. All diets were certified organic. The experiment was conducted during the grower and finisher phases in 4 different seasons: late fall, spring, summer, and early fall. Pasture access was assessed either by housing broilers on the West Virginia University Organic farm and giving them outdoor access, or by housing broilers on the West Virginia University Animal Sciences farm and giving them no outdoor access. Fast-growing genotypes were superior in performance and carcass characteristics compared with slow-growing genotypes, and choice-feeding management did not improve performance or carcass characteristics. Pasture access tended to have no effect on slow-growing broilers and decreased the performance of fast-growing broilers. Performance was decreased in late fall, likely because of cold ambient temperatures.     

Effects of dietary additives (potassium diformate/organic acids) as well as influences of grinding intensity (coarse/fine) of diets for weaned piglets experimentally infected with Salmonella Derby or Escherichia coli

The aim of this study was to examine whether and to what extent the addition of potassium diformate (pdf) or free organic acids (fpa) to the diet and the grinding intensity might affect the course of infection and the passage of orally applied Salmonella and Escherichia coli in pigs. Experiments were carried out using 80 reared piglets allotted to four groups. Pigs were fed pelleted diets ad libitum (except during a 15 h feed‐withholding‐period before infection). The control diet contained finely ground cereals (2 mm screen). To two test diets (also finely ground) 1.2% pdf, 0.9% organic acids (75% formic and 25% propionic acid, fpa) respectively were added. The fourth diet (without acids) was based on coarsely ground cereals (6‐mm screen). After experimental infection alternately with S. Derby or E. coli, the course of infection was examined (rectal swab technique). Pigs were sacrificed 4–5 h after a further oral application of ～10⁹–10¹⁰ CFU S. Derby or E. coli to determine the counts of Salmonella or E. coli in chyme (classical culture methods). Adding pdf or fpa to the diet led to reduced Salmonella shedding and resulted in significantly lower counts of Salmonella and E. coli in the stomach content indicating an improved efficacy of the stomach barrier. In the distal parts of the digestive tract, the effect was less obvious concerning counts of E. coli, whereas counts of Salmonella were reduced markedly as well. The diet based on coarsely ground cereals failed to demonstrate positive effects concerning infection and passage of orally applied bacteria as well, but this diet was also pelleted and showed unintentionally, comparable amounts of fine particles. Results obtained in this study allow the recommendation of using pdf or organic acids as additives when dietary measures against Salmonella or E. coli in pigs are required.     

载牛冠状病毒N蛋白壳聚糖微球的免疫效果评价

为了对壳聚糖分子作为佐剂的缓释效果进行免疫学评价,本研究合成了牛冠状病毒DB2毒株N蛋白基因的3'端第487位～1287位碱基,用大肠杆菌对其进行了高效表达,通过SDS-PAGE电泳分离、电洗脱纯化该牛冠状病毒N蛋白(BCV N).以戊二醛为交联剂,采用乳化交联的方法制备空白壳聚糖微球.利用吸附法制备载BCVN蛋白的壳聚糖微球,通过肌肉注射途径免疫BALB/c小鼠,用间接ELISA方法检测免疫鼠血清中的抗体水平,由此评价壳聚糖微球对BCVN的缓释作用和佐剂效应.结果表明,壳聚糖微球的形态较好、表面光滑、分散性较好,平均粒径为6.50±1.77μm,电势分布在36 mV,吸附BCV N蛋白的壳聚糖微球在体内所产生的免疫效果明显优于唯N蛋白组,说明载BCV N蛋白的壳聚糖微球在体内具有一定的缓释效果,使N蛋白在体内缓慢释放而长时间诱导抗体的产生,该研究结果为以壳聚糖微球作为疫苗佐剂积累了实验数据.     

鸡源株与人源株H9N2流感病毒血凝素受体结合位点氨基酸比较与分析

H19N2病毒可以感染多种禽类和哺乳动物，包括人类。流感病毒血凝素受体结合位点的氨基酸可以影响受体结合特性。为了解鸡源株与人源株H19N2病毒受体结合部氨基酸的差异，作者对二者进行了比较与分析，结果表明，鸡源株与人源株血凝素受体结合部位氨基酸的第137、183、190、226位点存在差异，鸡源株在这些位点分别为K、N、AorVorT、LorQ而人源株则分别为R、H、E、L。虽然尚不清楚这些位点的改变对病毒与细胞亲和力及宿主范围的影响，但由于H9N2在我国养鸡业的普遍存在，加强H9N2病毒的分子流行病学监测有重要意义。     

In vivo and in vitro efficacy of zoledronate for treating oral squamous cell carcinoma in cats

BACKGROUND: Feline oral squamous cell carcinoma (OSCC) may cause painful bone destruction. Given the local invasiveness and rapid clinical progression of OSCC, conventional therapies are often palliative. In human cancer patients, zoledronate exerts anticancer effects by inhibiting tumor-induced angiogenesis and malignant osteolysis. HYPOTHESIS: Zoledronate will exert in vitro and in vivo anti-angiogenic and antiresorptive effects in feline OSCC. ANIMALS: Eight cats with OSCC were prospectively treated with zoledronate and conventional treatment modalities. METHODS: In vitro, zoledronate's effects in modulating soluble vascular endothelial growth factor (VEGF) secretion and receptor activator of nuclear factor kappaB (NF-kappaB) ligand (RANKL) expression were investigated in a feline OSCC cell line (SCCF1). In vivo, basal serum C-telopeptide (CTx) concentrations were compared among normal and OSCC-bearing cats, and the biologic effects of zoledronate administration in cats with naturally occurring OSCC were quantified by serially assessing circulating serum VEGF and CTx concentrations. RESULTS: In vitro, zoledronate concentrations greater than 3 microM reduce soluble VEGF secretion in the SCCF1 cell line. The expression of RANKL in the SCCF1 cell line was also modulated by zoledronate, with low concentrations (3 microM) decreasing but higher concentrations (30 microM) increasing RANKL expression in comparison with untreated cells. In vivo, cats with bone-invasive OSCC had greater serum CTx concentrations in comparison with geriatric, healthy controls. Treatment with zoledronate rapidly decreased circulating serum VEGF and CTx concentrations in cats with spontaneously occurring OSCC. CONCLUSIONS AND CLINICAL IMPORTANCE: Zoledronate exerts in vitro and in vivo effects that may favor the slowing of tumor growth and pathologic bone turnover associated with OSCC.