用卡片PCR法检测卡耶他环孢子虫

使用FTA卡片法提取卡耶他环孢子虫DNA,利用特异性引物组F1E/R2B、CC719/CRP999进行巢式PCR,得到1条298bp的特异条带。并以柔嫩艾美尔球虫、贝氏隐孢子虫等作为对照,结果显示,除卡耶他环孢子虫外,均无特异性条带。用卡耶他环孢子虫卵囊进行巢式PCR敏感性试验,结果显示敏感度最高达1.0×100个/mL。     

浓缩大豆磷脂对肉仔鸡生产性能的影响

本试验采用单因子随机化设计,将225只艾维菌肉仔鸡随机分为5个处理组,每个处理组设3个重复,每个重复15只鸡,以玉米-豆粕型日粮为基础日粮,饲喂处理1设为空白对照组;处理2添加猪油,设为猪油对照组;处理3、4、5分别添加低、中、高水平的浓缩大豆磷脂,为试验组.在0～3周龄猪油以2%,浓缩大豆磷脂分别以2%、3%、4%;在4～6周龄猪油以3%,浓缩大豆磷脂分别以3%、4%、5%添加到基础日粮中以替代相同比例的玉米,蛋白质水平维持不变.研究添加猪油和不同水平的浓缩大豆磷脂对肉仔鸡生长性能的影响,并对其作用机理进行了初步探讨.结果表明:在0～3周龄时,中、高水平试验组极显著地提高了采食量(P<0.01);在4～6周龄时,中水平试验组具有较大的采食量,但高水平试验组采食量有所下降;从全期来看,猪油对采食量影响不大;在0～3周龄,中、高水平试验组极显著地提高了增重(P<0.01),而且在此阶段添加油脂显著提高了饲料转化率(P<0.05);在4～6周龄时,添加油脂极显著提高了增重和饲料转化率(P<0.01);从全期来看,添加油脂可以提高增重与饲料转化率,且在0～3周龄时浓缩大豆磷脂优于猪油.     

Application of PCR-HRM in the study of relationship between leptin gene promoter polymorphisms and liver cirrhosis

AIM: To analyze the feasibility of PCR-high-resolution melting curve analysis (HRM) for detecting the site mutations of C2549 and G2548 in leptin gene promoter from the patients with liver cirrhosis, and to explore the relevance between mutant genotypes and physiological and biochemical indexes in liver cirrhosis patients. METHODS: Compared with the method of PCR-restriction fragment length polymorphism (RFLP), the present research used the method of PCR-HRM to analyze the site mutations of C2549 and G2548 in leptin gene promoter in control group (n=100) and liver cirrhosis group (n=100). The physiological and biochemical indexes of the patients were also detected and compared. RESULTS: Leptin gene promoter polymorphism was detected using PCR-HRM with effectiveness, high flux and accuracy. Preliminary results showed that the main mutation of the patients with liver cirrhosis was in C2549 site, but not found in G2548 site. Leptin, free leptin index (FLI), fasting insulin (FINS) and insulin resistance index estimated by homeostatic model assessment (HOMA-IR) in liver cirrhosis group were higher than those in control group. Insulin sensitivity index (ISI) and soluble leptin receptor (sOB-R) in liver cirrhosis group were lower than those in control group with significant difference except leptin level. Meanwhile, FLI showed positively correlated with FINS and HOMA-IR (r=0.45, r=0.53, P<0.05), and negatively with ISI (r=-0.34, P<0.05). In the patients with liver cirrhosis, C2549A heterozygous mutation was predominant. The indexes of HOMI-IR, leptin, sOB-R and FLI of C2549A homozygotes and heterozygotes were higher than those of the wildtypes, which showed significant difference except leptin and sOB-R levels (P<0.05). CONCLUSION: PCR-HRM can be more accurate for identifying leptin promoter polymorphism. The increase in the frequency of C2549A mutation may be closely related with liver cirrhosis. Existence of hyperinsulinemia and insulin resistance may be correlated with leptin level in the patients with liver cirrhosis.     

生防菌XC-1的筛选、鉴定及其对马铃薯黑痣病的防效研究

从马铃薯根际土壤中分离筛选得到对立枯丝核菌有较强拮抗作用的菌株XC-1,对其进行鉴定及生防特性分析,并评价其对马铃薯黑痣病的生防效果.形态学、16S rDNA和gyrB基因序列分析以及生理生化检测结果表明,XC-1菌株为枯草芽孢杆菌(Bacillus subtilis).XC-1菌株具有产纤维素酶、蛋白酶及嗜铁素和HC...     

大黄鱼冷藏过程中的鲜度变化

研究了大黄鱼(Pseudosciaena crocea)贮藏于0℃(冰藏)、3℃、7℃和10℃过程中,在感官、化学和微生物品质方面的变化特性,并对其货架期进行分析,探讨了菌落总数、嗜冷菌、假单胞菌、产H2S菌、TVBN和TMA与感官评价的一致程度.结果表明,大黄鱼是海水鱼中鲜度下降较慢的种类,在O℃、3℃、7℃、10℃的贮藏过程中,大黄鱼的高品质期分别为330 h、208 h、114 h和87 h,货架期分别为523 h、368 h、197 h和150 h.各温度高品质期终点和货架期终点时菌落总数(CFU)/g的对数平均值分别为6.10±0.30和6.67±0.35,产H2S菌数(CFU/g)的对数平均值分别为5.82±0.33和6.37±0.22,TVBN均值分别为(14.77±0.5)mg/(100 g)和(28.02±1.19)mg/(100 g);TMA均值分别为(2.55+0.62)mg/(100 g)和(9.84±1.28)mg/(100 g),各温度下高品质期终点和货架期终点时各指标均值均无显著差异(P>0.05),表明菌落总数、产H2S菌数和TVBN值作为大黄鱼低温贮藏的鲜度指标与感官鲜度评价有较好的一致性.     

两种有机硅助剂对四氯虫酰胺防治菜青虫的增效作用

为明确两种有机硅助剂Silwet 408和Greenwet 7618对四氯虫酰胺防治菜青虫的增效作用,本文采用喷雾法测定四氯虫酰胺以推荐剂量、50%推荐用量分别与Silwet 408和Greenwet 7618的3 000倍稀释液混用对菜青虫的田间防效。结果表明,四氯虫酰胺以50%推荐用量分别与Silwet 408和Greenwet 7618混用,药后3 d对菜青虫的防效均与单独使用推荐剂量的防效无显著差异;药后7 d,50%推荐用量与Greenwet 7618混用对菜青虫的防效与推荐剂量的四氯虫酰胺仍无显著差异,而50%推荐用量与Silwet 408混用的防效显著降低;在药后14 d,所有处理对菜青虫无防治效果。因此,建议在生产上推广50%推荐用量的四氯虫酰胺与Greenwet 7618的3 000倍液混用防治菜青虫。     

滇西南菜区小菜蛾发生规律及抗药性现状

为掌握滇西南菜区小菜蛾的发生规律和抗药性现状, 2013-2017年进行了田间系统调查幼虫、性信息素诱集成虫和室内毒力测定。结果表明:滇西南菜区小菜蛾田间种群每年早春存在种群激增现象, 3-5月份出现春季高峰期, 6月后种群数量保持较低水平。抗性测定表明, 该地区小菜蛾对丁醚脲、Bt、氯虫苯甲酰胺、虫螨腈、茚虫威均为低水平抗性, 对氟啶脲和多杀霉素为中等水平抗性; 对阿维菌素为中到高抗,对高效氯氰菊酯为高抗水平。建议该区域1-5月放置性信息素诱杀成虫, 3-4月用丁醚脲、Bt制剂、氯虫苯甲酰胺、虫螨腈、茚虫威等进行化学防治, 6-12月小菜蛾的发生虫量在经济阈值下, 可不防治。     

Cloning and Tissue Expression Analysis of PDK4 and FGF10 Genes in Guizhou Congjiang Xiang Pig

The objective of this study was to clone PDK4 and FGF10 genes, and investigate the expression level of PDK4 and FGF10 genes mRNA in different tissues of Large White pig and Congjiang Xiang pig. The PDK4 and FGF10 genes were cloned by RT-PCR and analyzed by bioinformatics, the relative expression of PDK4 and FGF10 genes were detected by Real-time PCR. The results showed that the coding region of PDK4 gene was 1 224 bp, encoding 407 amino acids; The coding region of FGF10 gene was 636 bp and encoded 211 amino acids. The homologies of nucleotide sequences of PDK4 gene with sheep, horse and human were 93%, 92% and 91%,respectively. The homologies of nucleotide sequences of FGF10 gene with sheep, cattle, human and mouse were 94%,93%, 93% and 90%, respectively. The phylogenetic tree of PDK4 gene showed that the genetic relationship of Congjiang Xiang pig, cattle and sheep were very close, the phylogenetic tree of FGF10 gene indicated that the genetic relationship of Congjiang Xiang pig, cattle, sheep, human and macaque were very close, but the genetic relationship of Congjiang Xiang pig, rat and chicken were far away. Real-time PCR results showed that, in different tissues of Congjiang Xiang pig,PDK4 gene expression in kidney tissue was higher than other tissues, with a higher expression in stomach and adipose as well,FGF10 gene expression in stomach tissue was higher than other tissues, with a higher expression in kidney and adipose as well, but both of PDK4 and FGF10 genes expression were the lowest in longissimus dorsi. In different tissues of Large White pig, both of PDK4 and FGF10 genes were expressed the highest in adipose than other tissues, PDK4 gene expression in longissimus dorsi was the lowest, while the FGF10 gene expression the lowest in heart. This study successfully cloned the PDK4 and FGF10 genes of Large White pig and Congjiang Xiang pig,and detected the relative expression of PDK4 and FGF10 genes in different tissues of Large White pig and Congjiang Xiang pig, and also provided scientific basis for further study on regulation of PDK4 and FGF10 genes on lipid metabolism and deposition.     

苹果砧木M26试管苗不定根发育过程中蛋白质的变化

为进一步了解木本植物不定根发生、发育的机理,对苹果砧木M26试管苗不定根的发生、发育过程进行解剖观察,利用十二烷基磺酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)对M26试管苗在接入生根培养基前后,不同时期表达的蛋白进行了研究。结果发现:在不定根原基形成和发育的阶段性过程中,茎基部蛋白质的种类和数量出现明显差异。生根培养后,在继代苗茎基部表达的5条蛋白条带减弱或消失,其分子量分别为26.8 kDa、37kDa、40.3 kDa、43 kDa和66 kDa;明显表达了4条新的蛋白条带,其分子量分别为26 kDa、27.7 kDa、32.5 kDa、和45 kDa,同时有4条蛋白条带(38.5 kDa、42 kDa、53.2 kDa和55.5 kDa)其表达丰度提高。多种蛋白条带的消失和出现,可作为M26试管苗不定根产生的生化标志。在不定根原基逐步形成发育的同时,有3条蛋白条带的表达丰度也逐步提高,可作为不定根形成过程表达的特异标记蛋白带。