Sequence and Phylogenetic Analysis of rDNA ITS of Three Eimeria Species in Rabbit

In order to study whether the internal transcribed spacers (ITS) sequence could be used as a molecular marker for the species identification of rabbit coccidian, the rDNA ITS of Eimeria intestinalis, Eimeria flavescens and Eimeria magna were amplified by polymerase chain reaction (PCR), and were cloned into pGEM-T Easy vector subsequently. The positive recombinant plasmids were identified by PCR and then sequenced. By sequence comparison and comparative analysis with the relative sequences of rabbit Eimeria spp. available in GenBank, the results showed that the lengths of Eimeria intestinalis, Eimeria flavescens and Eimeria magna were 1065, 1009 and 1047 bp, respectively, and the sequence homologies with the same species sequences were 99.2%, 99.0% and 94.5%, respectively, while were 55.3% to 82.1% compared with corresponding sequences of other different species sequences. The phylogenetic analysis using software Mega 5.0 showed that all rabbit coccidia clustered together in a clade, which was divided into two sister lineages, corresponding to the presence or absence of oocyst residuum. The result demonstrated ITS could be used as a molecular marker for the species identification of rabbit coccidia.     

抗菌肽与抗生素的体外抗菌效果比较

应用由 Hancock实验室改进的微量肉汤稀释法测定了 cecropin P1、cecropin A、magainin 2、defensin 1、bactenecin、lactoferricin和 indolicidin等 7种抗菌肽和盐酸金霉素、去甲万古霉素、土霉素、强力霉素等 4种抗生素的体外抗菌活性 ,并且使用薄层琼脂糖孔穴扩散法比较了抗菌肽中 cecropin P1和 cecropin A与土霉素和呋喃唑酮对大肠杆菌 K88的抑菌效果。结果发现 ,抗菌肽与抗生素一样对几种革兰氏阳性菌和阴性菌都有不同程度的抗菌效果。其中 cecropin A、cecropin P1和 defensin1对大肠杆菌的 2个菌株 ATCC2 5 92 2和 K88的抗菌活性高于抗生素 ,defensin1是各种抗菌肽中对金黄色葡萄球菌抗菌效果最好的。抗生素对试验用金黄色葡萄球菌的抑制活性总体上要好于抗菌肽。与抗生素相比 ,cecropin A、cecropin P1和 indolicidin对猪霍乱沙门氏菌和鼠伤寒沙门氏菌具有更好的抗菌效果。另外 ,抗菌肽的抑菌圈边缘十分清晰 ,而抗生素的整个抑菌圈都比较模糊且界线不明。因此 ,从抗菌效果方面考虑 ,抗菌肽可以代替抗生素用于疾病的预防和治疗     

唇形科植物提取物对单胃动物的抗氧化作用及其机制

动物在患病、应激或特殊生理条件下,机体内会大量产生自由基,自由基与机体内的脂质、ＤＮＡ等产生反应,可对动物造成氧化损伤。唇形科植物提取物中的抗氧化物主要活性成分是酚类化合物,它和抗坏血酸及类胡萝卜素能够阻止自由基引发的氧化伤害。本文阐述了唇形科植物提取物对猪、鸡体外和体内抗氧化作用的实例与抗氧化功能,分析了其抗氧化作用机制。     

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。