我国牛源金黄色葡萄球菌耐药现状及药敏检测方法探讨

为调查我国牛源金黄色葡萄球菌的临床耐药现状,对2009年以来新疆、浙江、山东、内蒙古和上海五省区不同地方分离的120株金黄色葡萄球菌用纸片扩散法进行临床药敏试验。结果显示,新疆分离株对红霉素、克林霉素、青霉素、复方新诺明、多西环素、四环素耐药,22%的菌株对头孢西丁耐药,43%的对氯霉素耐药,64%的对环丙沙星耐药,58%的对庆大霉素耐药;其它4地区分离株耐药情况更为严重,除头孢西丁以外,对其它9种抗生素均耐药。对菌株的多重耐药情况分析发现,新疆分离株中对5种以上抗生素耐药的占86.1%,对10种抗生素完全耐药的占17.4%;而内地分离株的数据为100%和38.2%。此外,在试验中鉴定出33株耐甲氧西林金黄色葡萄球菌,新疆19株,占新疆菌株的22.1%,内地四地区14株,占内地分离株的41.2%。结合2005~2009年间发表的相关耐药数据的分析结果,研究显示,我国牛源金黄色葡萄球菌呈现多重耐药,且出现了耐甲氧西林金黄色葡萄球菌,并在不同地区分离株中占有一定的比例;研究还发现,新疆分离株与浙江、山东、内蒙古和上海四地分离株的耐药情况有较大差异。此外,在整理药敏试验资料时,发现在金黄色葡萄球菌药敏试验中,药物的选择没有遵循选药规则,试验操作不规范和试验结果的报告不规范等问题,对此提出一些探讨性建议,供试验人员参考。     

氨基脲人工抗原的合成及其多克隆抗体的制备

目的合成氨基脲人工抗原并制备其多克隆抗体。方法以氨基脲(SEM)和对醛基苯甲酸(CP)为原料合成半抗原(CP-SEM),并采用碳化二亚胺法和混合酸酐法将其分别与牛血清白蛋白(BSA)和鸡卵清蛋白(OVA)偶联,合成氨基脲人工抗原。以氨基脲人工抗原(CPSEM-BSA)为免疫原免疫Balb/c小鼠,利用间接ELISA法测定其抗体效价。结果经薄层层析、红外光谱和元素分析等方法鉴定合成半抗原即为CP-SEM。紫外扫描、聚丙烯酰胺凝胶电泳结果表明人工抗原合成成功。免疫获得抗氨基脲的多克隆抗体,其抗体效价为1:64000。结论氨基脲人工抗原合成成功,并获得抗氨基脲的多克隆抗体。     

The Productive Performance and the Correlation Analysis Between Hatchability and Egg Quality in Jiangxi Green-eggshell Layers

The purpose of this study was to provide the reference to Jiangxi Green-eggshell breeding through the analysis of laying performance, hatchability and egg quality in the first generation of Jiangxi Green-eggshell layers at 300-days old. 306 Jiangxi Green-eggshell layers and 80 eggs were selected to determine the laying performance, hatchability and egg quality among families, and the correlation between the hatchability and egg quality were analyzed also. The results were as fellows:The average body weight of these hens was 1 596.99 g, the average egg weight was 50.57 g, the average egg-laying number was 14.53, the average fertility rate was 93.69%, the average dead fertile egg rate was 5.55%, the average healthy chick rate was 84.3%, the average dead embryo egg rate was 13.9%, the average egg shape index was 1.28, the average Haugh unit was 71.77, and the average eggshell strength was 4.44 kg/cm². The healthy chick rate and the egg weight had significant correlation,as well as the eggshell color and the egg shape index (P < 0.05). The healthy chick rate had extremely significantly correlation with the dead fertile egg rate(P < 0.01), as well as the eggshell strength and the eggshell thickness (P < 0.01). Generally, the first generation of Jiangxi Green-eggshell layers had good egg quality, higher laying performance and hatchability, but they still need to be further selected.     

Effects of Photoperiod on Stem Cells Activation in Cashmere Goat Hair Follicles

In order to extend the anagen of cashmere goat hair follicles and increase the production of cashmere,this study was performed with artificially shorten the daylight time among Arbas White cashmere goats. Skin tissue sections from cashmere goats were collected to compare the morphologic changes between artificial daylight and natural daylight,and immunohistochemical method was used to study the hair follicle cell proliferation and important protein expression in related signaling pathways. The results showed that strong cell proliferation occurred in cashmere goat hair follicle cells during artificial daylight,plenty of cytokeratin 15 (K15) positive signals were distributed in the outer root sheath,β-catenin protein was actively expressed in hair matrix and root sheath, indicating that the hair follicles were in the anagen growth phase;Meanwhile,cashmere goat hair follicles under natural daylight were in telogen with weak signals. Above all prove that short photoperiod played an important role in promoting hair follicle growth,the artificial short photoperiod could change hair follicle growth cycle and make hair follicles earlier enter to the anagen growth phase,causing a variety of typical gene expressions during hair follicle growth.     

刺五加提取物抗仔猪断奶应激的效用

【目的】研究刺五加提取物(ASE)的抗仔猪断奶应激作用。【方法】生长试验选用96头体重相近的21日龄断奶仔猪,按体重和性别随机分为3个处理,每个处理4个重复,分别饲喂基础日粮、添加ASE日粮和添加抗生素日粮。试验期21d,观察仔猪生长性能,测定血清生化参数及相关激素水平的变化。消化试验选用6头仔公猪(同上),按2×3×3复拉丁方设计,饲喂上述3种日粮,测定营养物质的消化率。【结果】与基础日粮组相比,ASE可显著提高日增重、显著降低料肉比;提高日粮钙、总能和磷的表观消化率;提高断奶后血清低密度脂蛋白、胆固醇和葡萄糖的含量,降低血清尿素氮含量并提高血清碱性磷酸酶活性;显著提高血清游离甲状腺素T3和T4、生长激素及胰岛素样生长因子-1水平和T3/T4。【结论】ASE可调节激素分泌,提高断奶仔猪消化代谢水平,从而减缓断奶应激,促进仔猪生长。     

生物秸秆对肉鸡表观代谢能的影响及替代玉米适宜比例的研究

本试验旨在研究肉鸡对生物秸秆的表观代谢能及其替代玉米的适宜比例.生物秸秆代谢能的测定试验选择24只42日龄健康的爱拔益加(AA)肉鸡,随机分为3个处理,每个处理8只鸡.处理1饲喂含8％生物秸秆的基础饲粮,处理2饲喂以30％的生物秸秆代替基础饲粮中能量饲料和蛋白质饲料的饲粮,处理3采用强饲法进行测定.生物秸秆代替玉米的试验选择150只42日龄健康的AA肉鸡,随机分为5个处理,每个处理5个重复,每个重复6只鸡.对照组饲喂基础饲粮,试验组分别在基础饲粮中用4％、8％、12％生物秸秆代替玉米(代谢能按照差量法计算结果12.46 MJ/kg计)及8％生物秸秆代替玉米(代谢能按照强饲法计算结果5.30 MJ/kg计,添加豆油,使其代谢能水平与对照组相同).结果表明:1)基础饲粮的代谢能为12.63 MJ/kg,采用差量法推算出肉鸡对生物秸秆的表观代谢能为12.46 MJ/kg,而采用套算法和强饲法测定生物秸秆的表观代谢能分别为4.38和5.30 MJ/kg.2)与对照组相比,4％和8％的生物秸秆代替玉米组中性洗涤纤维和酸性洗涤纤维的代谢率显著升高(P＜0.05),而12％的生物秸秆代替玉米组表观能量的代谢率显著降低(P＜0.05).由此得出,套算法和强饲法低估了生物秸秆的表观代谢能,而差量法更能准确反映生物秸秆的表观代谢能;在肉鸡饲粮中生物秸秆可以替代4％ ～8％的玉米.     

牛病毒性腹泻病毒单克隆抗体捕获ELISA方法的建立

以纯化的牛病毒性腹泻病毒(BVDV)作为抗原免疫BALB/C系小鼠,通过细胞融合和克隆筛选出稳定分泌BVDV抗体的杂交瘤细胞株,将杂交瘤细胞接种于BALB/C系小鼠的腹腔内诱生腹水,腹水单抗与BVDV均呈阳性反应.将5A9、2G3和5C2(分别结合不同的抗原结合位点)诱生的BVDV单抗纯化后等量混合包被酶标板,与鸡抗BVDV IgY(检测抗体)联合应用,建立BVDV抗原捕捉ELISA(AC-ELISA)方法.采用方阵滴定法确定单抗、鸡抗BVDV IgY的最适工作浓度,用阴性组织样品作为判定检测结果的D490临界值,最后以建立的AC-ELISA检测临床粪样棉拭子27份,结果表明:该方法敏感性、特异性高,与全血病毒分离结果的符合率迭86.36%;与全血和粪样RT-PCR检测结果的符合率分别为89.47%和94.74%.阴性对照RT-PCR、病毒分离和AC-ELISA检测均为阴性.     

日粮不同蛋白质水平对三元肥育猪生产性能和胴体品质的影响

本文旨在研究不同日粮蛋白质水平对三元肥育猪生产性能、血液参数、胴体性状和肉质性状的影响.试验选取36头杜长大三元杂交猪,体重约为(47±4.2)kg,随机分为3个处理,每个处理12个重复,各处理分别饲喂低(12.9%)、中(15.4%)、高(18.7%)3个蛋白质水平日粮.试验第46天后,分别检测血液生化指标、激素水平、胴体品质和肉质性状.结果显示,3个日粮蛋白质水平对日增重、采食量、料重比及胴体品质和肉质性状均无影响(P>0.05);血清碱性磷酸酶、低密度脂蛋白、甘油三酯、胆固醇、谷丙转氨酶、谷草转氨酶、血清尿素氮以及生长激素均以中蛋白质水平组最高;随着日粮蛋白质水平的增加,肥育猪血清胰高血糖素有降低的趋势(P=0.089).以上结果表明,日粮蛋白质水平降低到12.9%并不影响肥育猪生长性能、胴体品质和肉质性状.     

牛病毒性腹泻病毒新疆分离株E2基因的克隆及序列分析

应用RT-PCR方法从牛病毒性腹泻病毒（BVDV）新疆石河子分离株、玛纳斯不同年份两个分离株扩增获得的3株BVDV的E2基因序列,分别命名Shihezi148、Manasi和MNS2（GenBank登录号：EU159699、EU159703和EU169937）。序列分析结果表明3株E2基因长度分别为1121bp、1122bp和1122bp,分别编码373、374和374个氨基酸残基。核酸序列同源性和系统发生分析表明,3株病毒均属于BVDV基因1型（BVDV-1）,Shihezi 148株与澳大利亚Bega株亲缘关系最近,同处于BVDV-1c基因亚型群,其E2氨基酸同源性为90.11%。Manasi株和MNS2株E2基因核酸序列仅有4个碱基的差别,它们与Changchun 184和匈牙利VEDEVAC株亲缘关系最近,位于BVDV-1b基因亚型群。Manasi和MNS2株与Changchun 184株E2氨基酸同源性分别高达98.66%和98.93%。     

大米糖浆替代乳糖对早期断奶仔猪生长性能及胰岛素样生长因子轴的影响

本文旨在研究大米糖浆取代乳糖后对早期断奶仔猪生产性能及胰岛素样生长因子轴的影响.选用7 d断奶的二元杂交仔猪(长白×大白)32头,随机分为2个处理组,每组设16个重复,每个重复1只.日粮成分除乳糖水平外(乳糖组和大米糖浆组分别为43.5%和0),其他成分均一致.试验期14 d,在试验第7天、第14天和第21天对试验猪进行空腹8 h称重,计算试验期日增重及日耗料,并分别于试验第7天和第14天每组随机抽取5头,颈静脉采血,屠宰速取肝脏和背最长肌组织.结果表明,大米糖浆取代乳糖后对仔猪的日增重及肉料比均没有显著影响(P>0.05),14 d大米糖浆组血清GH水平与乳糖组差异不显著(P>0.05),21 d大米糖浆组血清GH水平极显著低于乳糖组(P<0.01);但大米糖浆组14 d和21 d血清IGF-Ⅰ的水平,及肝脏和背最长肌IGF-Ⅰ mRNA相对表达量与乳糖组差异均不显著(P>0.05).综合分析认为,大米糖浆替代乳糖后对早期断奶仔猪的生长性能和胰岛素样生长因子轴没有影响.