Effect of sulphide in Pacific white shrimp Penaeus vannamei under varying oxygen and pH levels

Sulphide is one of the known environmental stressors, which potentially binds to cytochrome C oxidase (COX), a key enzyme in the electron transport chain, thereby blocking oxygen transport and ATP production. To ascertain the toxic effects of sulphide on Pacific white shrimp, Penaeus vannamei, two distinct exposure experiments were carried out with varying concentration of sulphide (0–1 mg/L), dissolved oxygen (normoxia and hypoxia) and pH values (8.2, 6 and 5). The activity of enzymes viz COX, superoxide dismutase (SOD), phenoloxidase and lactate accumulation was investigated. Outer membrane integrity and COX monomer separation were also done with the isolated crude mitochondrial preparations. Results indicated a significant reduction (p ≤ .05) in COX enzyme activity in sulphide exposed treatments when compared to control. The reduction was more intense when pH levels were reduced under hypoxia condition. Lactate accumulation as a result of anaerobic metabolism was found to be higher in hypoxic treatments. No significant difference (p ≥ .05) was observed in superoxide dismutase activity between the treatments, whereas phenoloxidase activity significantly decreased at higher concentration sulphide. Separation of mitochondrial proteins resulted in the identification of ~205 kDa of COX monomer, and significant damage was found in outer membrane integrity under hypoxia and pH treatments. From this study, it is evident that at a given concentration, sulphide is toxic to P. vannamei, and in association with hypoxia and low pH, they further intensify sulphide toxicity. Our results indicated that sulphide toxicity should not be considered as a single factor, rather it should be a considered as combination of factors.     

Impact of different beer yeasts on wheat dough and bread quality parameters

In order to investigate the impact of different yeast strains from the species Saccharomyces cerevisiae on the dough and bread quality parameters, wheat flour was fermented using different beer yeasts. The results show that beer yeast strains could be included in the baking process since S. cerevisiae T-58 and S. cerevisiae s-23 provided adequate gas production and dough formation with superior structural properties like extensibility and stickiness to S. cerevisiae baker's yeast. The resulting breads show the highest specific volume with the highest slice area and the highest number of cells and the lowest hardness over time. The different yeasts had also an impact on the crust colour due to their abilities to ferment different sugars and on shelf life due to the production of a range of different metabolic by-products. According to this study it was possible to produce higher quality bread by using yeast coming from the brewing industry, instead of bread containing standard baker's yeast.     

恩诺沙星诱导水生细菌产生耐药性最低浓度选择

渔药多以拌料或直接投入水体的方式给药,药物经动物排泄最终进入土壤和表层水体中,造成药物在表层水体等环境中残留,药物在环境中残留将诱导环境细菌产生耐药性,而且耐药性可能通过食物链、环境或动物和人直接接触进行传递。为了解恩诺沙星在池塘水体中残留对水生细菌的影响,采用琼脂稀释法对水生细菌进行了抗菌药物敏感性试验。结果表明,恩诺沙星诱导水生细菌产生耐药性的最低浓度选择为8μg／mL。     

复方恩诺沙星可溶性粉对畜禽主要致病菌的体外抑菌效果

采用纸片扩散法(K-B法)测定复方恩诺沙星可溶性粉对畜禽主要致病菌的敏感性,采用试管倍比稀释法测定复方恩诺沙星可溶性粉对畜禽主要致病菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)。结果表明,病原性大肠埃希菌、绿脓杆菌、金黄色葡萄球菌、鸡巴氏杆菌、猪巴氏杆菌和链球菌对复方恩诺沙星可溶性粉敏感,复方恩诺沙星可溶性粉对畜禽主要致病菌的MIC为0.0279μg/mL~3.575μg/mL,MBC为0.055μg/mL~7.11μg/mL。     

Development of a real‐time PCR assay for detection and quantification of Streptococcus iniae using the lactate permease gene

The aim of this study is the development and evaluation of a rapid and accurate quantitative PCR (qPCR)‐based protocol for detection of zoonotic pathogen Streptococcus iniae in bacterial cultures and tissues of diseased fish. For this purpose, the lactate permease‐encoding (lldY) gene was selected as a target for the design of S. iniae‐specific primers based on comparative genomic analysis using 45 sequences retrieved from NCBI genome database. Specificity and applicability of these primers were tested using 115 bacterial strains and fish tissues infected with S. iniae. Sensitivity, reproducibility and efficiency of qPCR assay were also determined. The developed qPCR assay showed 100% specificity with pure bacterial cultures or DNA extracted from S. iniae or tissues of fish infected with the bacterium. The method has high sensitivity with a detection limit of 1.12 × 10¹ amplicon copies per assay (equivalent to 2 × 10^–9 ng/µl) using bacterial DNA and of 1.44 × 10¹ gene copies in tissues of fish infected with S. iniae. In conclusion, this qPCR protocol provides an accurate and sensitive alternative for the identification of S. iniae and its detection on fish tissues that can be implemented as a routine tool in microbiological laboratories.     

低浓度恩诺沙星对离体恒化器模型中人体肠道菌群的影响

 【目的】应用离体恒化器模型评价低浓度恩诺沙星对人体肠道菌群的影响,为制定微生物学日允许摄入量（ADI）提供依据。【方法】四套接种人体肠道菌群的离体恒化器模型中连续添加0、0.2、2及20 mg•L-1恩诺沙星8 d,采用细菌培养计数和PCR-DGGE方法研究细菌数量、耐药性、菌群结构和定植抗力的变化。【结果】2及20 mg•L-1恩诺沙星可抑制肠道总厌氧菌、总需氧菌和兼性厌氧菌、大肠杆菌、肠球菌的生长（P＜0.05或P＜0.01）;0.2、2及20 mg•L-1恩诺沙星均使总需氧和兼性厌氧菌、大肠杆菌耐药百分率增加（P＜0.05或P＜0.01）, 停药7 d后,总需氧和兼性厌氧菌、大肠杆菌耐药百分率呈下降趋势,但与对照组相比仍处于较高水平（P＜0.01）。20 mg•L-1恩诺沙星对肠道菌群结构的影响较大,使PCR-DGGE图谱发生明显变化,条带数、菌群多态性和相似性参数降低（P＜0.05或P＜0.01）;2和20 mg•L-1恩诺沙星可使肠道菌群对外源沙门氏菌的定植抗力明显下降。【结论】0.2 mg•L-1恩诺沙星（相当于3.67μg•kg-1体重）可对离体恒化器模型中的人体肠道菌群耐药性产生显著影响,说明中国现行规定的恩诺沙星和环丙沙星的ADI（6.2μg•kg-1体重）可能会对人体肠道菌群产生影响,主要为选择出耐药需氧和兼性厌氧菌。     

恩诺沙星及其代谢产物在奶牛乳中的消除规律

 【目的】研究乳房炎奶牛单剂量乳房灌注1%恩诺沙星（50 ml/头）后乳中药物浓度的变化规律。【方法】采用HPLC法测定乳中恩诺沙星及其代谢产物环丙沙星的浓度,用统计矩原理处理药物浓度—时间数据。【结果】在给药区恩诺沙星和代谢产物环丙沙星均于给药后2ｈ左右乳中浓度达到最高值,分别为（13.16±3.10）μg?ml-1和（2.79±0.94）μg?ml-1,然后逐渐下降;12 ｈ后检测不到恩诺沙星,24 ｈ已检测不到环丙沙星。对于非给药区,恩诺沙星给药后1ｈ左右乳中浓度达到最高值（0.19±0.02）μg?ml-1,然后逐渐下降;6 ｈ后检测不到恩诺沙星,而代谢产物环丙沙星于给药后4 ｈ乳药浓度可达最高峰（0.51±0.07）μg?ml-1,24 ｈ已检测不到乳药浓度。给药区代谢物环丙沙星的消除半衰期较恩诺沙星长,平均为（1.67±0.20）h,平均滞留时间（MRT）亦长。在本试验条件下,建议临床休药期不少于3 d。【结论】恩诺沙星吸收较迅速 ,代谢较快、清除较快 ,表明恩诺沙星与环丙沙星在乳中不会形成残留。     

基于P-糖蛋白基因表达评价尼罗罗非鱼体内恩诺沙星代谢“首过效应”

为从药物酶的角度建立一种客观评价鱼类"首过效应"的方法,利用荧光定量PCR法测定尼罗罗非鱼(Oreochomis niloticus Linn)肝、肾组织中P-糖蛋白(P-gp)基因表达量,分析了单剂量(40 mg.kg 1)口服给药恩诺沙星(enrofloxacin,ENR)后,尼罗罗非鱼肠道、肝组织中mRNA水平的相对表达量与ENR血药浓度的时实相关性。实验结果显示:在尼罗罗非鱼肠道、肝组织中,P-gp基因在分子量127 bp处出现了与预期大小相符的特异性扩增片段。对尼罗罗非鱼口灌给药ENR后,ENR能迅速通过肠道进入血浆,其在肠道、肝和血浆中的消除速度较快,其药物时量曲线关系符合一级吸收的二室开放动力学模型。当血浆中ENR浓度达到最高达峰时(1 h),实验组肠道和肝中P-gp基因的相对表达量相对于对照组均表现出显著性差异(P<0.05);当肠道中ENR浓度达到最高峰时(2 h),实验组肠道P-gp基因的相对表达量则表现出极显著差异(P<0.01);当肝中ENR浓度达到最高峰时(2 h),实验组肝P-gp基因的相对表达量与对照组相比则表现出显著性差异(P<0.05)。该结果证实了鱼类P-gp基因参与药物代谢过程,提供了一种从分子水平揭示水产动物体内药物代谢规律的思路。     

不同产地人参中4种脱氢酶活力比较

为了给人参的培育和优选提供理论依据,采用中性缓冲液提取粗酶液,应用分光光度法对15个不同产地的人参中苹果酸脱氢酶(Malate Dehydrogenase MDH),乳酸脱氢酶(Lactate Dehydrogenase, LDH),乙醇脱氢酶(Alcohol Dehydrogenase, ADH),葡萄糖-6-磷酸脱氢酶(Glucose-6-phosphate Dehydrogenase, G6PDH）4种脱氢酶活力进行比较。结果表明不同产地人参脱氢酶活力差别明显,同一产地4种脱氢酶活力趋势基本相同。其中安图县万宝镇的人参样品的MDH、LDH、G6PDH 3种酶活力均是最高值,分别为124.58U/(g?Fw)、129.88 U/(g?Fw)、109.84U/(g?Fw);黑龙江2个产地的4种酶活力普遍比较低。运用SPSS13.0软件对15批样品进行系统聚类分析,将样品分为4类。MDH、LDH、ADH、G6PDH的活力可以作为人参培育和优选提供理论依据。