液相色谱-串联质谱法测定猪血浆和尿液中8种同化激素的残留

为建立猪血浆和尿液中8种同化激素的液相色谱-串联质谱(LC-MS/MS)检测方法,样品酶解后,用乙酸乙酯超声提取,经C18固相萃取小柱净化,进行LC-MS/MS检测。结果表明,采用基质添加标准曲线消除基质干扰,8种同化激素在1.5μg/L~100μg/L内,相关系数均大于0.99,线性关系良好。除炔诺酮定量限为1.5μg/L,其他7种药物定量限均为1μg/L。在2、10、50μg/L添加水平下,药物的回收率为71.5%~92.3%,日内相对标准偏差(RSD)为2.2%~14.6%,日间RSD为3.2%~12.7%。本方法灵敏度高,适用于同化激素类药物残留的检测分析。     

The Effect of Gender on Slaughtering Performance and Meat Quality in Sunite Sheep

The purpose of this experiment was to study the effect of gender on the slaughter performance and meat quality of Sunite sheep.Twelve 6-month-old Sunite rams and ewes at the same body condition were selected from the grazing herds of the Sunite Zuoqi Improvement Station Breeding Farm to determine their slaughter performance and meat quality.The slaughter test results showed that the bone quality and bone-meat ratio of the rams were significantly higher than that of the ewes (P<0.05),and the net meat quality and net meat rate of the ewes were significantly higher than that of the rams (P<0.05).The results of meat quality traits showed that the drip loss of ewes were significantly lower than that of rams (P<0.05),and the other indicators were not significantly different (P>0.05).The results of conventional nutrient composition of muscles showed that the intramuscular moisture content of the rams were significantly higher than that of the ewes (P<0.05),and the intramuscular fat content of the ewes were significantly higher than that of the rams (P<0.05),while the other indicators were not significantly different (P>0.05).The results of amino acid determination showed that the essential amino acid ratio in muscle of sunite ewes were significantly higher than that of rams (P<0.05),but there was no significant difference between the rams and ewes that formed the precursor amino acid content of meat flavor (P>0.05).Fatty acid measurement results showed that the saturated fatty acid ratio in the subcutaneous fat of rams was significantly lower than that of ewes (P<0.05).There was no significant difference in the ratio of saturated fatty acids in muscle fat and tail fat (P>0.05);The ratio of monounsaturated fatty acids in ewes tail fat was significantly higher than that of rams (P<0.05).The ratio of monounsaturated fatty acids in subcutaneous fat and intermuscular fat was not significantly different between rams and ewes (P>0.05).The ratio of polyunsaturated fatty acids in intermuscular fat of rams was significantly higher than that of ewes (P<0.05),and there was no significant difference in the content of polyunsaturated fatty acids in subcutaneous fat and tail fat (P>0.05).In summary,it showed that gender had a certain influence on the slaughter performance and meat quality of 6-month-old Sunite sheep.     

Prokaryotic Expression of Brucella BAB Gene and Establishment of iELISA

In this study,the genome of Brucella Rev.1 strain was used as a template to amplify the BAB gene sequence,clone it into the prokaryotic expression vector pET-30a(+),obtain the recombinant plasmid pET-30a-BAB,and perform prokaryotic expression on the recombinant plasmid.The indirect ELISA method was constructed by using the expression products detected by Western blotting.The results showed that the BAB gene was successfully cloned and expressed in this experiment,and the purified expression product was analyzed by SDS-PAGE.This study obtained a relatively pure recombinant BAB protein;Western blotting test showed that the expressed protein could react specifically with Brucella sheep positive serum and had good reactogenicity;Using the recombinant BAB protein as the coating antigen,an indirect ELISA method for detecting BAB antibodies was established and optimized.The best determined coating conditions were as follows BAB protein coating amount was 0.25 μg/mL,serum dilution was 1:400;blocking solution was 3% pig-derived gelatin;secondary antibody dilution was 1:6 000;color development time was 10 min.The established method was used to detect 40 clinical sheep serums,and the cut-off value was calculated to be 0.607.That was,when the serum tested had P/N ≥ 1.5 and D_{450 nm} ≥ 0.607,it was judged as positive,when D_{450 nm} ≤ 0.561,it was judged as negative,and when 0.607<D_{450 nm}<0.561,it was judged as a suspect value,and retest was required.Compared with the Huhong plate test and the test tube agglutination test,the positive coincidence rate was 100%,the negative coincidence rate was 71.88%,and the total coincidence rate was 77.5%.     

非淀粉多糖酶对奥尼罗非鱼生长及抗菌力的影响

本文旨在研究非淀粉多糖酶对奥尼罗非鱼的生长及其抗菌力的影响。试验选用体重约60 g的奥尼罗非鱼(Oreochromis niloticus×O.aureus)560尾,随机分成4个处理组,每组7个重复,每重复20尾奥尼罗非鱼,分别投喂添加不同梯度的液体酶制剂(0、0.1、0.2和0.4 mL/kg的饲料)。结果表明:(1)非淀粉多糖酶对罗非鱼的特定生长率、饲料转化率等生产性能指标均有显著升高(P<0.05);(2)添加非淀粉多糖酶0.2 mL/kg和0.4 mL/kg显著提高了罗非鱼白细胞的吞噬活性(P<0.05)和血清的杀菌能力(P<0.01),试验组各组织溶菌酶活力也显著增加(P<0.05)。     

固始鸡、罗曼蛋雏鸡和艾维茵肉仔鸡生长、养分沉积、肉质特性的比较研究

通过研究固始鸡、罗曼蛋雏鸡和艾维茵肉仔鸡生长、养分沉积、肉质特性的差异,探讨品种与生长、代谢和肉质间的关系。选用1日龄雄性固始鸡、艾维茵肉仔鸡、罗曼蛋雏鸡各120只,按体重随机分为6个重复,每重复20只,在相同日粮条件下进行8周的生长试验,并进行屠宰试验和肉质测定。结果表明:(1)3种鸡在体重、日增重、采食量、饲料转化率间存在极显著差异(P<0.01),固始鸡低于罗曼蛋雏鸡,罗曼蛋雏鸡低于艾维茵肉仔鸡;(2)固始鸡和罗曼蛋雏鸡的代谢能和粗蛋白质沉积效率显著低于艾维茵肉仔鸡(P<0.05);(3)固始鸡和罗曼蛋雏鸡的肌肉滴水损失、肌纤维直径、羟脯氨酸含量显著低于艾维茵肉仔鸡(P<0.05),固始鸡胸肌和腿肌中超氧化物歧化酶活性则极显著高于蛋鸡(P<0.01),蛋鸡极显著高于肉鸡(P<0.01)。与快速生长的鸡品种相比,慢速生长的固始鸡具有较低的代谢能和粗蛋白质沉积效率,具有较低的肌肉滴水损失、羟脯氨酸含量及较细的肌纤维,并具有较高的肌肉抗氧化水平。     

藏鸡和隐性白羽鸡感染柔嫩艾美耳球虫后免疫相关基因的表达变化分析

试验旨在从免疫遗传学的角度初步探讨藏鸡（TC）和隐性白羽鸡（RWC）对柔嫩艾美耳球虫（Eimeria tenella）易感性差异的分子机制,分别用1×10⁵个柔嫩艾美耳球虫孢子化卵囊感染对球虫具有抗性的藏鸡和易感的隐性白羽鸡。应用实时荧光定量PCR检测藏鸡和隐性白羽鸡感染前0 d和感染后第2、4、6和8天脾脏、盲肠、胸腺、法氏囊中γ-干扰素（IFN-γ）、白细胞介素-2（IL-2）、IL-16、Toll样受体（TLR3）和TLR15免疫相关基因的转录水平变化。结果显示,藏鸡脾脏IFN-γ、IL-2、IL-16及TLR3、TLR15免疫相关基因转录水平于感染后第4和8天明显上调,隐性白羽鸡则无明显变化。藏鸡盲肠IFN-γ转录水平在感染后第2天显著上调（P<0.05）,TLR3在感染后第4天起显著上调（P<0.05）,其余免疫相关基因变化幅度不大;隐性白羽鸡盲肠IFN-γ转录水平在感染后第8天显著上调（P<0.05）,IL-2在感染后第2天起显著上调（P<0.05）,IL-16在感染后第6天起显著上调（P<0.05）,TLR3在感染后第2和8天显著上调（P<0.05）,TLR15变化幅度不大。各免疫相关基因在2个品种鸡胸腺和法氏囊中均出现上调或下调,但除藏鸡法氏囊TLR3和TLR15转录水平变化幅度相对较大外,其余免疫相关基因与感染前相比变化幅度不大。以上结果显示,球虫感染主要导致藏鸡和隐性白羽鸡脾脏和盲肠中的各免疫相关基因出现显著变化,表明宿主的遗传背景在一定程度上可影响球虫感染的免疫应答。     

球虫消杀剂（Oo-stat）的配方优化研究

为获得对鸡球虫卵囊有良好抑杀作用的球虫消杀剂最佳配方,将3种不同配方的球虫消杀剂配成1:20、1:30、1:40、1:50、1:60、1:80、1:100、1:150、1:200、1:300及1:400.分别对柔嫩艾美耳球虫未孢子化卵囊体外相互作用6h、12h和24h之后,培养观察其孢子化率.结果表明,配方一、配方二除1:300和1:400处理6h组及1:400处理12h组与对照组差异不显著外,其余各组均显著低于生理盐水对照组(P<0.05);1:20、1:30、1:40、1:50、1:60及1:80的球虫消杀剂溶液分别处理卵囊6h、12h和24h后孢子化率均低于5%;配方三1:20、1:30、1:40、1:50、1:60及1:80的球虫消杀剂溶液分别处理卵囊12h和24h后孢子化率均低于5%,但是当处理时间为6h时,其孢子化率较高.研究表明配方二具有优良的抑杀鸡球虫卵囊的作用.     

甘肃中部地区禾谷镰孢的变异研究

为掌握禾谷镰孢在甘肃中部地区的分布及变异情况,从根部表现有坏死和叶鞘发褐的小麦幼苗的不同部位、小麦地土壤及玉米籽粒、玉米茎秆上分离禾谷镰孢,并以形态学为基础,参照Nelson分类系统进行鉴定。结果表明,在分离到的43个镰刀菌菌株中,有14个菌株经鉴定为禾谷镰孢,均从玉米茎秆上分离到,小麦根部、小麦叶鞘、小麦地土壤、玉米籽粒中分离到的镰刀菌中未见禾谷镰孢。将禾谷镰孢在特定条件下培养后,发现14个禾谷镰孢菌株产生子囊壳的数量不同,为2～90个。在以Fg16为引物的PCR反应中,随机选取的11个禾谷镰孢菌株都产生0.41kb的PCR产物,而6个对照菌株都产生0.50kb的片段,证明引物Fg16可以区分禾谷镰孢菌株群体的遗传多态性。以Tri13为引物的PCR反应显示,11个禾谷镰孢菌株以及3个中国对照菌株都产生脱氧雪腐镰刀菌烯醇(DON)毒素,而3个澳大利亚对照菌株产生雪腐镰刀菌烯醇(NIV)毒素。     

用RAPD标记分析部分柞蚕二化性品种资源的遗传多样性

采用RAPD标记技术,对36份柞蚕(Antheraea pernyi)二化性品种资源的遗传多样性进行分析。PCR扩增及琼脂糖凝胶电泳结果表明:利用筛选出的19个随机引物在供试材料中共检测到123个信息位点,其中多态性位点114个,多态性比率92.68%;每一个引物可检测出3～9个位点,其多态性比率不尽相同,平均扩增出6.47条带;各品种具有不同数量的特异性带型。供试品种资源的Nei氏遗传多样性指数(h)为0.324 3±0.151 5,Shannon信息指数(I)为0.487 1±0.197 5,表明长期同地保存的二化性柞蚕品种资源同样具有丰富的遗传多样性;供试品种的Nei氏遗传一致性范围在0.495 9～0.837 4之间,达0.6以上的占96.59%,表明多数品种资源的相似程度较高。36份品种资源分别以辽宁省蚕业科学研究所育成种为主和以国内外引进种为主聚为两大类群。     

鸭疫里默氏菌病和大肠杆菌病多重PCR诊断方法的建立

参考GenBank中鸭疫里默氏菌和大肠杆菌的外膜蛋白A(OmpA)基因序列,应用PrimerPremier5.0软件在二者高度保守区设计了2对引物,建立了适合鸭疫里默氏菌和大肠杆菌的快速检测的多重PCR检测方法。以该方法对已分离并保存的鸭疫里默氏菌和大肠杆菌进行PCR扩增,分别扩增出与试验设计相符的670、408bp的特异性DNA片段。将扩增所得的DNA片段进行克隆测序,测序结果表明分别为鸭疫里默氏菌和大肠杆菌OmpA基因序列。该方法对鸭疫里默氏菌和大肠杆菌的检测下限分别为4×104CFU/mL和3×104CFU/mL。表明所建立的PCR方法具有特异、快速和敏感的特点,可用于诊断鸭疫里默氏菌、大肠杆菌以及两者的混合感染。