高效液相色谱法测定猪血浆及组织中喹赛多及其脱二氧代谢物

建立了测定猪血浆及肝脏、肾脏、肌肉等组织中的喹赛多及其代谢物脱二氧喹赛多的高效液相色谱(HPLC)法.血浆样用甲醇沉淀蛋白后离心取上清液进样测定.组织样先用乙腈匀浆,用正己烷脱脂后作HPLC检测.色谱柱为ODS C18柱;流动相血浆样测定为乙腈水(2080),组织样测定为甲醇水(4258),流速为1.0 mL/min;紫外检测波长305 nm.药物工作液质量浓度范围为0.005～0.500 mg/L时,血浆样品及组织样品测定条件下药物质量浓度与响应值均具良好线性关系,相关系数＞0.999.血浆中药物质量浓度为0.02、0.10、0.50 mg/L时,喹赛多及脱二氧喹赛多的回收率均大于70%,组织中药物含量为0.05、0.20、1.00μg/g时,肌肉样品的回收率均大于70%,肝脏、肾脏样品则为50%～80%.本试验条件下,喹赛多及其脱二氧代谢物的最低检出质量浓度,血浆样品分别为0.01、0.02mg/L,组织样品2种检测物均为0.025μg/g.测定了工作液3种质量浓度0.01、0.05、0.25 mg/L的仪器精密度,日内相对偏差＜8.0%,日间相对偏差＜17.0%.     

Mutant-prevention concentrations of enrofloxacin for Escherichia coli isolates from chickens

OBJECTIVE: To investigate the development of enrofloxacin resistance among Escherichia coli isolates obtained from chickens by determining mutant-prevention concentrations (MPCs) and sequence the quinolone resistance-determining regions (QRDRs) of gyrA and parC genes in selected isolates. SAMPLE POPULATION: 15 chicken-derived E coli isolates. PROCEDURES: For all isolates, MPC and minimal inhibition concentration (MIC) of enrofloxacin were determined. The MPCs and maximum serum drug concentrations attained with enrofloxacin doses recommended for treatment of E coli infections in chickens were compared. Mutation frequencies and QRDR sequence changes in gyrA and parC were also determined. RESULTS: In 2 of 15 E coli strains, MPCs were low (0.016 and 0.062 microg/mL), MPC:MIC ratios were 2 and 4, and the GyrA and ParC proteins had no mutations. In 9 susceptible isolates with a GyrA point mutation, MPCs ranged from 2 to 16 microg/mL. For isolates with double mutations in GyrA and a single mutation in ParC, MPCs were > 32 microg/mL (several fold greater than the maximal plasma concentration of enrofloxacin in chickens); mutation frequencies were also much lower, compared with frequencies for single-mutation isolates. CONCLUSIONS AND CLINICAL RELEVANCE: For E coli infections of chickens, MPC appears to be useful for determining enrofloxacin-dosing strategies. The high MPC:MIC ratio may result in enrofloxacin-treatment failure in chickens infected with some wild-type gyrA E coli isolates despite the isolates' enrofloxacin susceptibility (MICs 0.125 to 1 microg/mL). For infections involving isolates with high MPCs, especially those containing mutations in gyrA and parC genes, treatment with combinations of antimicrobials should be adopted.     

对奶牛胎衣不下的认识和处理

本文结合本场的生产实际，讨论了母牛胎盘的特点，胎衣不下的症状与诊断；着重讨论了本病的发病率及其影响因素，并将病因分析、归纳成一个示意图；最后综述了多项治疗和预防胎衣不下的措施，如灌服羊水，注射激素，用维生素A、D、E和亚硒酸钠，灌服中药，手术剥离以及电刺激等。     

蚕桑业主的兴起给城郊农村发展蚕业的启示

翠屏区地处宜宾市的经济文化中心地区,农村经济以近郊蔬菜、远郊水果为主.蚕桑有一席之地,但有日渐萎缩趋势.蚕业发展的经验和教训告诉我们,在这样的环境中发展蚕桑,需要探索引进切合实际的新路子.实践表明,"组合养蚕",用重点突破、以点带动发展的的方式让蚕业在远郊农村形成一村一品、一社一业的格局,是一条有希望的路子."业主或准业主"模式就是一种很好的"组合养蚕"方式.     

Identification of polymorphisms and association analysis with meat quality traits in the porcine KIAA1717 and HUMMLC2B genes

Skeletal muscle genes are potential candidates for production and meat quality. Screening a subtracted cDNA library constructed with mRNA obtained from longissimus dorsi muscles of F1 hybrids Landrace × Yorkshire and their female parents Yorkshire, we isolated two partial sequences coding for the H3-K4-specific methyltransferase (KIAA1717) and skeletal muscle myosin regulatory light chain (HUMMLC2B) genes. Database search revealed KIAA1717 and HUMMLC2B encoded proteins with SET domain and EF-hand calcium binding motif, respectively. In the present work we identified their partial polymorphisms and two SNPs, one (C1354T) at the 3′ untranslated region (UTR) of KIAA1717 and one (A345G) at the SINE (PRE-1) element of HUMMLC2B, both created/disrupted a restriction site for endonuclease Msp I. The selected pigs were genotyped at the KIAA1717 C1354T and HUMMLC2B A345G sites by means of a PCR-RFLP protocol. Significant associations were observed for the KIAA1717 C1354T polymorphic site with meat marbling (longissimus doris (p < 0.05), biceps femoris (p < 0.01)) and intramuscular fat (p < 0.01). HUMMLC2B A345G were significantly associated with meat pH (longissimus doris (p < 0.05), biceps femoris (p < 0.01)), drip loss (p < 0.01), water holding capacity (p < 0.01) and meat color value (longissimus doris (p < 0.01), biceps femoris (p < 0.05)). Further studies are needed to confirm these preliminary results.     

Use of comparative proteomics to identify the effects of creatine pyruvate on lipid and protein metabolism in broiler chickens

Four hundred male chickens were selected to study the effects of pyruvate (Pyr), creatine pyruvate (CrPyr) and creatine (Cr) on the expression of hepatic mitochondrial and cytoplasm proteins associated with lipid and protein metabolism. Mitochondrial purification was accomplished using the two-step differential centrifugation and density gradient method, and the activities of organelle-specific marker enzymes were determined to assess the purity of the mitochondria. Proteins were extracted and fractionated by two-dimensional electrophoresis and the differential protein spots were assessed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. CrPyr reduced fatty acid accumulation by down-regulating adipose differentiation-related protein, inhibited ATP synthase expression, and reduced cholesteryl ester transfer protein (CETP) expression, thus reducing the levels of high density lipoprotein and triglycerol (TG) levels (thereby lowering fat and cholesterol deposition). CrPyr increased the expression of eukaryotic translation initiation factor (eIF) 2B, calreticulin (CRT) and eIF3a, thus promoting protein synthesis. CrPyr up-regulated the expression of fatty acid-binding proteins, CETP and apolipoprotein A-IV in cytoplasmic extracts, and these proteins accelerated the decomposition of fatty acids and TG, thus reducing fat deposition. In conclusion, CrPyr plays an important role in lipolysis and protein synthesis, and this effect was more pronounced than was the effect of Pyr and Cr.     

犬瘟热、犬细小病毒胶体金二联检测卡研制

本研究通过细胞融合、间接ELISA方法筛选出4株可稳定分泌犬瘟热病毒(canine distemper virus,CDV)和犬细小病毒(canine parvovirus,CPV)单克隆抗体的杂交瘤细胞株,以此为基础研制了一种可同时检测CDV和CPV的胶体金二联检测卡,并对其特异性、灵敏度及准确性进行了测试。结果显示：4株杂交瘤细胞可稳定传代,分泌的单克隆抗体纯度高,效价均在1:320以上。制备的胶体金二联检测卡特异性强,只与CDV和CPV产生特异性条带,而不与其他犬类病毒反应;灵敏度高,最低检测限可达到10²拷贝/μL;准确性好,与荧光定量PCR结果的符合率高于88.0%,与市面上单一病毒检测卡的符合率高于97.2%。结果表明,本研究制备的CDV、CPV胶体金二联检测卡特异性强、灵敏度高、准确性好,兼具操作简便、肉眼可判读、结果易保存和无需特殊仪器设备等优势,可用于两种疾病的临床快速诊断和大规模检测工作。     

Evaluation of the colonization capabilities of Salmonella Enteritidis in Quails using an RT-PCR approach

We used a real-time PCR assay and indirect fluorescent antibody (IFA) assay to detect genomic DNA of Salmonella Enteritidis in the internal organs of quails after an oral challenge. The results showed that S. Enteritidis was detected in all the samples at different time points. This study will assist a future understanding of the pathogenesis of S. Enteritidis.     

蒽酮-硫酸比色法检测多糖条件的优化

本研究通过单因素实验和L9(33)正交实验优化蒽酮-硫酸比色法检测多糖含量的条件,研究反应温度、反应时间、蒽酮-硫酸液的体积对吸光度值的影响。结果表明,蒽酮-硫酸比色法检测的糖含量的最优条件为蒽酮-硫酸2 mL,于100℃反应10 min。验证实验结果表明,总标糖获得的标准曲线方程A=0.005W-0.0162,r=0.9998,优于葡萄糖标准曲线方程。在620 nm下测定,能得到较高的吸光度值,重现性好。     

不同生长阶段群体数量特征对产草量作用的分析

在相同的光，温，水，肥条件下，同时对营养生长期的高羊茅和生殖生长期的多年生黑麦草人工草地产草量和群体数量特征的定量关系进行研究。在营养生长期，参数的变异程度小于生殖生长期。营养生长期各因子对产草量直接作用的程度排序这：叶面积指数〉草丛高度〉分蘖密度，生殖生长期为；草丛高度〉叶面积指数〉分蘖密度。