猪、犬寄生虫的检验诊断方法

在现代养猪、犬的生产中,寄生虫病是造成饲料报酬低,影响养猪、犬经济效益的一个重要因素。但在饲料中寄生虫的危害没有引起养殖户的足够重视,给养殖户造成严重的经济损失。1寄生虫分体内寄生虫和体外寄生虫两大类1.1体内寄生虫体内寄生虫主要有蛔虫、弓形体病、囊虫病、旋毛虫病、肺丝虫病等,这几种体内寄生虫对猪、犬机体危害较大,成虫与猪、犬争夺营养成分,移行虫体破坏猪、犬的肠壁、肝脏和肺脏的组织结构和生理机能，造成Et增重减少，抗病力下降，怀孕母猪胎儿发育不良，甚至造成隐性流产，新生仔猪小和窝产仔猪数少等。     

动物寄生虫病的分类、诊断与防治

1分类寄生虫是暂时或永久寄居于另一种生物的体表或体内，夺取被寄居者的营养物质，并给被寄居者造成不同程度危害的动物。寄生虫种类繁多，根据其与宿主的关系，可分为专性寄生虫、兼性寄生虫、体内寄生虫、体外寄生虫和机会致病寄生虫。     

响应面法优化仙茅多糖酶解工艺及体外免疫活性

采用葡聚糖酶对仙茅多糖XMP-1进行酶解,并利用响应面法对酶解工艺进行合理优化。在单因素试验的基础上,采用星点设计模拟葡聚糖酶酶添加量、酶解温度、时间、pH值等4个因素对还原糖生成量综合影响的模型,得到最佳工艺条件,并验证模型的可靠性。通过小鼠巨噬细胞(RAW264.7)分析XMP-1 1～12 h酶解得到的12批产物的体外免疫活性。结果表明,最佳酶解工艺为酶添加量为56%,温度为52℃,酶解时间为12 h,pH值为6.0,此时还原糖生成量为0.201 0 mg/mL。优化结果可靠,与预测值的相对误差小于5%。不同时间段酶解产物对细胞无明显毒害作用,均能诱导RAW264.7细胞活化,释放TNF-α和NO,其中酶解5 h的效果最好。该研究可为仙茅多糖酶降解片段的制备提供理论基础,也可为仙茅多糖在食品领域进一步开发和应用提供科学依据。     

亚洲棉EST-SNP的挖掘及其在陆地棉中的验证

【目的】随着不同棉种序列数据库的逐步完善以及高通量测序技术的发展,棉花单核苷酸多态性(Single nucleotide polymorphism,SNP)标记开发可利用的公共数据资源逐步增加。【方法】本研究基于陆地棉祖先基因组的现代种亚洲棉表达序列标签(Expressed sequence tag,EST)数据库,利用CAP3对亚洲棉EST数据库进行拼接。拼接获得7 187个重叠群(Contig),再利用Quality SNP软件进行SNP位点分析。【结果】在807条含有4条以上EST序列的Contig中查找到2 690个SNP位点。通过筛选次要等位基因频率大于30%的位点,获得953个可靠度较高的候选SNP,通过电子筛选,最终获得可用于陆地棉分析的SNP 149个,利用位点特异性聚合酶链式反应以及酶切扩增多态序列验证了EST-SNP的准确性。【结论】本研究证实基于亚洲棉EST数据库挖掘用于陆地棉研究的EST-SNP切实可行,并有望将EST-SNP用于陆地棉遗传图谱构建、重要性状的基因定位以及分子标记辅助育种。     

脂滴在Beagle犬卵母细胞体外成熟中的研究

犬排出的卵母细胞处于GV期,其体外成熟率在20%左右,尚未获得突破,阻碍了犬体外保种及基因修饰疾病模型创制研究。犬卵母细胞含有大量脂滴,关于脂滴与卵母细胞成熟的研究鲜有报道。综述了脂滴与犬卵母细胞体外成熟的关系,尝试寻找研究犬卵母细胞体外成熟新的突破口。     

Effects of Adding Mannan Oligosaccharides to Diets with Different Concentrate to Forage Ratios on Ruminal Fermentation of Sheep in vitro

The test was aimed at determining the effects of adding MOS to diets with different concentrate to forage ratios on ruminal fermentation of sheep in vitro.The 4×6 two-factor experimental design was chosen.There were 4 diets with different concentrate to forage ratios(20:80, 30:70, 40:60 and 50:50) and added to 6 dosages MOS(0, 0.4%, 0.8%, 1.2%, 1.6% and 2.0%), respectively.The method of gas production in vitro was used to measure the total gas and CH₄ production, IVDMD, IVCPD and IVOMD after cultured for 3, 6, 9, 12 and 24 h.The results showed that the gas(except 12 and 24 h) and CH₄ production(except 9 h) were not impacted by the two factors(P>0.05).The IVDMD, IVCPD and IVOMD were significantly affected by concentrate to forage ratios(P<0.05), while the IVCPD at 24 h was significantly affected by MOS(P<0.05).Meanwhile, all the indexes were not influenced by the interaction of two factors(P>0.05).The IVDMD and IVOMD increased with the rising of concentrate approximately, while the IVCPD decreased with the rising of MOS roughly.The biggest associative effect was gained in 30:70 concentrate to forage diet when adding 1.2% MOS.     

Effect of Human Menopausal Gonadotropin,17β-estradiol and Epidermal Growth Factor on the Quality of in vitro Maturation Bovine Oocytes

For optimizing in vitro maturation system of bovine oocytes,we firstly examined the influence of four different hormonal regimes(FSH+LH,HMG,FSH+LH+E₂ and HMG+E₂) on oocyte maturation rates.Then we studied the effects of epidermal growth factor (EGF) in the above defined medium on bovine oocyte maturation,in vitro development and quality of parthenogenetic embryos.The cell apoptotic index of parthenogenetic blastocysts was detected by TUNEL.No significant difference was observed in maturation rates in four groups supplemented with different hormones.However,human menopausal gonadotropin (HMG) provided steady maturation results in replicates.Maturation of oocytes was promoted by supplementation with 17β-estradiol (E₂).Combination of HMG and E₂ gave rise to steady and efficient mature results.The presence of EGF at 30 ng/mL concentration significantly increased maturation rate and blastocyst rate and reduced apoptotic cells in parthenogenetic blastocysts.Therefore,the optimal oocyte maturation solution could be supplemented with 0.075 IU/mL HMG,1 μg/mL E₂ and 30 ng/mL EGF.     

MCM-41型介孔纳米材料对氧氟沙星的负载及体外释放

虽然氧氟沙星抗菌谱广,抗菌作用强,在人医及兽医临床上被广泛地应用,但是其生物半衰期短,给临床用药带来不便。为了开发长效氧氟沙星制剂,本研究以高比表面积、比孔容的MCM-41型介孔二氧化硅纳米材料为载体,制得氧氟沙星纳米药物OFX@MSN。并分别通过扫描电镜、透射电镜,氮气吸附-脱附、傅里叶红外等手段对载药前后的纳米粒结构进行了结构确证,证明氧氟沙星载入到介孔二氧化硅的孔洞中形成平均直径约为100nm的分散球状粒子。体外试验显示氧氟沙星的释放时间可以持续3d之久,从而有效维持平稳的血药治疗浓度,减小注射次数。     

植物乳杆菌NJAU-01体外抗氧化活性的研究

为更全面和客观地评价植物乳杆菌NJAU-01的体外抗氧化活性,以商品化鼠李糖乳杆菌LGG为阳性对照,植物乳杆菌JMZ-12为阴性对照,对其体外自由基清除能力进行比较;采用电化学快速检测技术进一步评价乳酸菌缓解H_2O_2诱导巨噬细胞RAW264.7氧化损伤的能力。结果表明:10~9 CFU·mL~(-1) NJAU-01无细胞提取物的DPPH自由基清除率、羟自由基清除率以及超氧阴离子自由基清除率和还原能力分别为24.20%、29.03%、18.21%和40.13μmol·L~(-1)L-半胱氨酸,其自由基清除能力与阳性对照LGG相当。电化学结果显示NJAU-01无细胞提取物孵育后RAW264.7抗氧化能力最强,且胞内活性氧水平最低。这一研究提示, NJAU-01无细胞提取物是该菌株抗氧化能力最强的活性组分,可作为进一步解析乳酸菌抗氧化机制的研究基础。