副溶血性弧菌vtrB基因敲除菌株的构建及其在抵抗胆汁中的作用

构建副溶血性弧菌转录调节因子VtrB （V.parahaemolyticus T3SS2 regulator B）敲除菌株和回补菌株,探讨VtrB在副溶血性弧菌抵抗胆汁中的作用。根据GenBank数据库中副溶血性弧菌vtrB基因上下游序列设计引物,通过融合PCR将vtrB基因上下游同源臂融合并克隆入自杀质粒pDS132中,将重组自杀质粒转入大肠杆菌S17-1 λpir中,再接合转移至副溶血性弧菌中进行同源重组,通过10%蔗糖筛选获得vtrB基因突变株ΔvtrB。将pBAD33-vtrB重组质粒电转入ΔvtrB突变株中获得回补菌株C-ΔvtrB。研究副溶血性弧菌野生型、突变株和回补株在2%脱氧胆酸盐（DOC）处理后的存活率。基于同源重组原理,经vtrB基因的PCR扩增和转录水平检测结果表明,成功获得副溶血性弧菌vtrB基因缺失突变株ΔvtrB和回补株C-ΔvtrB。与野生型和回补菌株C-ΔvtrB相比,ΔvtrB突变株在2% DOC处理20、40和60 min后的存活率分别为17%、4%和1%,vtrB基因的失活使得副溶血性弧菌对胆汁抵抗能力极显著降低（P<0.001）,表现出存活能力的缺陷。由此表明,转录调节因子VtrB有助于副溶血性弧菌对胆汁的抵抗。     

动物福利视野下我国野生动物保护法的革新——从“活熊取胆”事件谈起

动物福利是指给予动物满足其康乐的外部条件.从动物福利的角度检视,我国的野生动物保护法存在对虐待、伤害动物等行为规制的缺失、野生动物栖息地保护不足等问题,“活熊取胆事件”即是最好的例证.建议我国的野生动物保护法借鉴日本、韩国、德国等国家的做法,吸纳动物福利保护的理念,构建多元化的野生动物福利保护制度,严格规制虐待野生动物的行为,加强对野生动物及生物多样性的保护.     

Cloning of Bile Salt Hydrolase Gene and Its Expression in Lactic Acid Bacteria

According to the sequence of the bile salt hydrolase (BSH) gene of Bifidobacterium and the restriction enzyme cutting sites of expression vector pNZ8148, primers were designed and the bile salt hydrolase (BSH) gene was gotten from Bacillus bifidus ATCC 29521 by PCR. BSH gene was inserted into lactic acid bacteria expression vector pNZ8148 to construct the recombinant pNZ8148-BSH. The recombinant pNZ8148-BSH was transferred into lactic acid bacteria NZ9000 with electrotransformation method. And the recombina...     

胆汁酸添加剂对草鱼抗氧化能力的影响

选取体长、体质量相近的草(Ctenopharyngodon idellus)240尾.随机分为4个组.每组3个重复,每个重复20尾.在Ⅰ、Ⅱ、Ⅲ、Ⅳ组的日粮中分别添加0、75、150、300 ms/ks胆汁酸制剂,试验期60 d.在30 d和60 d时,分别检测了血清和肝脏的超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、谷胱甘肽还原酶(GR).结果显示,在草鱼日粮中添加胆汁酸制剂,可以提高草鱼血清和肝脏的SOD、GSH-Px、GR活力,其中,以150 mg/kg组的水平最高,但各组间均无显著差异;另外,血清和肝脏的GR活力没有随胆汁酸添加量的增加而呈现规律性变化,但试验组的GR活力均高于对照组.以上结果表明,在饲粮中添加适宜剂量的胆汁酸制剂,能增强草鱼血清和肝脏的抗氧化能力,有可能改善鱼体生长状况.     

Sensitivity and specificity of fasting ammonia and serum bile acids in the diagnosis of portosystemic shunts in dogs and cats

Background: Portosystemic shunt (PSS) is the most common cause of hepatic encephalopathy in dogs and cats. Fasting ammonia and serum bile acids (SBA) are used to diagnose PSS, but their true sensitivity and specificity have not been fully evaluated, especially in cats. Objectives: The purpose of this study was to determine the diagnostic accuracy of fasting ammonia and SBA concentrations in the diagnosis of PSS in dogs and cats and to compare diagnostic accuracy between species. Methods: A retrospective analysis of data from 373 dogs and 85 cats presented to the clinic from 1996 to 2006 was carried out. Based on clinical, laboratory, and imaging findings, animals were grouped as having PSS, parenchymal hepatic disease, or extrahepatic disease. The sensitivity and specificity of ammonia and SBA concentrations for the diagnosis of PSS were calculated and receiver‐operating characteristic analysis was used to optimize cut‐offs. Results: Using the upper limit of laboratory reference intervals (ammonia, 59 μmol/L; SBA, 20 μmol/L), the sensitivity and specificity of ammonia was 85% and 86% in dogs, and 83% and 76% in cats, respectively. The sensitivity and specificity of SBA was 93% and 67% in dogs, and 100% and 71% in cats, respectively. Using optimal cut‐off points for ammonia (dogs, 57 μmol/L; cats, 94 μmol/L) the sensitivity and specificity was 91% and 84% in dogs and 83% and 86% in cats, respectively. Using optimal cut‐off points for SBA (dogs, 58 μmol/L; cats, 34 μmol/L) the sensitivity and specificity was 78% and 87% in dogs and 100% and 84% in cats. Conclusion: Increased fasting ammonia and SBA concentrations are accurate indicators of PSS. An improvement in diagnostic accuracy can be achieved by using defined optimal cut‐off points for the selective diagnosis of PSS.     

16S rDNA文库法分析草鱼肝胰脏和胆汁细菌群落组成

利用16S rDNA文库随机测序法分析了草鱼肝胰脏和胆汁细菌菌落组成,结果表明,在肝胰脏中共鉴定出19种共生细菌,分别隶属于气单胞菌属、假单胞菌属、摩根菌属、不动杆菌属、弧菌属和变形杆菌属。气单胞菌属和假单胞菌属为优势属,气单胞菌属的丰度为36.17%,假单胞菌属为31.91%。其中,气单胞菌属的GZ16和假单胞菌属的GZ9菌株丰度最高,均为12.77%。在胆汁中共鉴定出10种共生细菌,分别隶属于气单胞菌属、假单胞菌属、摩根菌属和变形杆菌属。气单胞菌属和假单胞菌属为优势属,气单胞菌属的丰度为35%,假单胞菌属为27.5%。其中,变形杆菌属的DN1菌株丰度最高,为25%。肝胰脏中只有10.5%的细菌群落与胆汁的细菌群落相似,胆汁中只有20%的细菌群落与肝胰脏相似,表明草鱼肝胰脏和胆汁有各自特有的共生细菌群落。本研究结果,为进一步探讨肝胰脏和胆汁共生细菌的生理功能,以及细菌与宿主草鱼之间的协同进化提供了基础。     

Dried green leaf powders of Jew's mellow (Corchorus), persimmon (Diosphyros kaki) and sweet potato (Ipomoea batatas poir) lower hepatic cholesterol concentration and increase fecal bile acid excretion in rats fed a cholesterol-free diet

A cholesterol-free diet containing dried powder of Jew's mellow leaves, persimmon leaves or sweet potato leaves respectively at 5% level as dietary fiber was fed to male Sprague-Dawley rats for about one month. The experiment was conducted twice except for sweet potato leaves. In the groups fed the diet mixed with powders of any of the three different dried green leaves, the hepatic cholesterol concentration significantly decreased. Such lowering was not observed in serum cholesterol concentration compared with the control (cellulose) group. A significant increase in fecal weight was observed in all the groups fed the green leaf samples. All the dried green leaves increased fecal excretion of bile acids per gram or per day compared with the control group in both experiments, but only the dried Jew's mellow leaves showed an increased excretion of neutral sterols. These results suggest that lowering of hepatic cholesterol by powdered green leaves is not necessarily due to the same factor, but to the increased fecal excretion of bile acids due to inhibited enterohepatic circulation in animals given these samples.     

胆汁酸对齐口裂腹鱼幼鱼脂肪沉积、脂肪代谢酶活性及相关基因表达的影响

为探讨胆汁酸对齐口裂腹鱼幼鱼生长、脂肪沉积、脂肪代谢及相关基因表达的影响;实验以360尾初始体质量为(12.74±0.14) g的健康齐口裂腹鱼幼鱼为对象,随机分为4组,每组设3个重复组,每个重复组放养30尾,分别投喂胆汁酸含量为0、75、150和300mg/kg的4种饲料,养殖时间为70 d。结果显示,随着胆汁酸含量的升高,齐口裂腹鱼的增重率(WGR)呈先升高后降低的变化趋势,且在胆汁酸含量为150 mg/kg时达到最大,为226.63%;其肠脂肪酶(LPS)、肝酯酶(HL)、脂蛋白脂酶(LPL)和总脂酶(TL)活性均呈先升高后趋于稳定的变化趋势;而脂肪酸合成酶(FAS)活性则呈相反的变化趋势;齐口裂腹鱼肝脏LPL mRNA表达量呈先上调后趋于稳定的变化趋势,FAS mRNA表达量呈先下调后趋于稳定的变化趋势;肝脏、肌肉及全鱼中粗脂肪含量均随胆汁酸含量的升高呈先降低后趋于稳定的变化趋势,胆汁酸含量对实验鱼的成活率及脂肪沉积效率无显著性影响。研究表明,本实验条件下,添加适量的胆汁酸可有效上调齐口裂腹鱼LPL mRNA的表达量,下调FAS mRNA的表达量,增强脂肪代谢酶活性,促进对饲料...     

高脂饲料中胆汁酸水平对齐口裂腹鱼肠道组织结构及脂肪代谢酶活性的影响

为探索高脂饲料中添加胆汁酸对齐口裂腹鱼幼鱼肠道组织结构及几种脂肪代谢酶活性的影响,以360尾初始体质量为(12.74±0.14)g的健康齐口裂腹鱼幼鱼为实验对象,随机分为4组,每组3个重复,每个重复30尾实验鱼,分别投喂添加了0、75、150和300mg/kg胆汁酸的4种实验饲料,养殖时间为70 d。结果显示,胆汁酸浓度从0增大到75 mg/kg时,齐口裂腹鱼前肠和中肠管壁厚度及肠绒毛高度显著升高,前肠皱襞宽度显著降低。而当胆汁酸浓度再进一步增大时,实验鱼前肠和中肠管壁厚度及肠绒毛高度则升高不明显,其皱襞宽度也降低不显著。未添加胆汁酸的实验组中齐口裂腹鱼前肠和中肠纹状缘畸形或脱落现象严重,随着胆汁酸添加量的增加,逐渐减少至消失。齐口裂腹鱼肠道中脂肪酶(LPS)及肝胰脏中脂蛋白酯酶(LPL)、肝酯酶(HL)和总酯酶(TL)活性均随胆汁酸添加量的增加而呈先升高后趋于稳定的变化趋势。且LPS、LPL活性均在胆汁酸添加量为300 mg/kg时最强(分别为2881.17 U/g和14.43 U/mg prot);HL、TL活性则在胆汁酸添加量为150 mg/kg时活性最强(分别为43.70和58.03 U/mg prot)。但LPS、LPL、HL和TL活性在150与300mg/kg组均无显著差异。研究表明,在饲料中添加75~150 mg/kg的外源胆汁酸能提高齐口裂腹鱼幼鱼脂肪代谢酶的活性,促进对饲料脂肪的代谢和利用,还能改善齐口裂腹鱼幼鱼肠道结构,保护肠道健康。     

大豆黄酮、脱脂乳对猪源芽孢乳杆菌S1耐受酸和胆盐的影响

通过体外实验研究了大豆黄酮和脱脂乳对芽孢乳杆菌S1在不同pH值和不同浓度胆盐下存活能力的影响。结果表明, 与对照组相比, 添加 10mg·mL-1的脱脂乳显著提高了S1在pH分别为 2 6、3 7条件下的存活数量, 并且也能显著提高S1在 1、2和 3mg·mL-1胆盐浓度条件下的存活数量, 说明脱脂乳对S1有显著的保护作用。添加 50mg·L-1的大豆黄酮也显著提高S1耐胆盐的能力, 但对S1耐受酸能力有负效应。