噁喹酸在凡纳滨对虾体内药动学和对弧菌的体外药效

采用高效液相色谱法,研究了复方噁喹酸粉药饵投喂在凡纳滨对虾体内药动学和组织中消除规律,同时检测了噁喹酸对对虾源弧菌的最小抑菌浓度(MIC),建立了药动/药效(PK/PD)关系,提出了用药方案和休药期建议。结果显示,复方噁喹酸粉拌饵投喂给药,噁喹酸给药剂量为30 mg/kg(体质量),凡纳滨对虾血浆噁喹酸浓度—时间关系曲线均符合一级吸收二室开放动力学模型。血淋巴中噁喹酸达峰浓度(Cmax)、达峰时间(Tmax)、曲线下面积(AUC0-24)和消除半衰期(t1/2z)分别为14.70 mg/L、2 h、244.6 mg/(L·h)和18.56 h;肌肉、肝胰腺和鳃的峰浓度(Cmax)分别为4.11、17.20和7.01 mg/kg,消除半衰期(t1/2z)分别为10.71、12.31和16.75 h。噁喹酸对132株弧菌的MIC主要分布在0.15~1.25μg/m L,MIC50和MIC90分别为0.62和1.25μg/m L。PK/PD相互关系参数Cmax/MIC90和AUC0-24/MIC90分别为11.76和195.7。研究表明,噁喹酸以30 mg/(kg体质量)剂量药饵给药,凡纳滨对虾能很好地吸收噁喹酸,可以有效地防治弧菌引起的细菌性疾病。     

噁喹酸和氟甲喹在鳗鱼体内的药代动力学研究

研究了单次血管注射喹酸(Oxolinicacid)和氟甲喹(Flumequine)后,在鳗鱼(Muraenesoxcinereus)体内的代谢规律及残留情况。采用高效液相色谱-质谱联用(HPLC-MS)法进行喹酸和氟甲喹的检测,检测低限(LOD)1ng/ml;定量限(LOQ)5ng/ml。喹酸和氟甲喹的代谢过程均可以用一级代谢二室模型来描述,喹酸的分布半衰期:0·18h,消除半衰期:14·84h,总体清除率0·07L/kg·h;氟甲喹的分布半衰期:0·19h,消除半衰期:9·38h,总体清除率0·07L/kg·h。两种药物在鳗鱼体内都有很强的“首过效应”,易在胆汁中积累;给药72h后,喹酸和氟甲喹在可食用组织肌肉中的残留量分别为36·1ng/g和21·8ng/g,低于目前国外要求的最高残留量50ng/g和75ng/g。     

Multiple dose pharmacokinetic study of oxolinic acid in cod, Gadus morhua L.

The plasma, muscle and liver distribution and elimination of the antibacterial agent oxolinic acid were studied after multiple oral (p.o.) administration of 10 or 20 mg kg⁻¹ day⁻¹ to cod (Gadus morhua) for 6 days. The fish, held in seawater at 6 and 12°C and weighing 150–250 g were sampled 24 h following last medication. The concentrations in plasma and tissues were clearly dosage and temperature dependent. The distribution from plasma to muscle (muscle/plasma ratio) was higher than that from a single dose study and independent of temperature and dosage. The distribution from plasma to liver (liver/plasma ratio) was lower than the muscle/plasma ratio and according to this study dependent of the administered dosage but independent of temperature. The elimination of oxolinic acid from plasma, muscle and liver was considerably faster following multiple administration compared to a single administration.     

Dietary folic acid requirement determined for grass shrimp, Penaeus monodon

A feeding trial was conducted to determine the dietary folic acid requirement of juvenile grass shrimp, Penaeus monodon. Purified basal diets were formulated using vitamin-free casein as the protein source. Graded levels (0, 1, 2, 5, 10, 15, 30, and 60 mg folic acid/kg diet) of folic acid were added to the basal diet, resulting in eight dietary treatments in the experiment. Each diet was fed to three replicate groups of shrimp initially averaging 0.79±0.01 g for 8 weeks. Shrimp fed diets supplemented with 2 mg folic acid/kg diet had greater (P<0.05) weight gain and hepatopancreatic folic acid concentrations than shrimp fed the unsupplemented control diet. Feed efficiency (FE) was highest in shrimp fed the diets supplemented with 2 mg folic acid/kg diet, followed by the groups fed 1 mg/kg, then the control group. Hepatosomatic index (HSI) was highest in shrimp fed the control diet, followed by 1 mg folic acid/kg and lowest in shrimp fed diets supplemented with 2 mg folic acid/kg. Weight gain percentage, hepatopancreatic folic acid concentration and hepatosomatic index of the shrimp analyzed by broken-line regression indicated that the adequate dietary folic acid concentration in growing P. monodon is 1.9–2.1 mg/kg diet.     

Development of hemolymph prostaglandins assay systems and their concentration variations during ovarian maturation in the kuruma prawn, Penaeus japonicus

Hemolymph prostaglandin (PGF₂ and PGE₂) measurement systems for the kuruma prawn Penaeus japonicus were developed and validated. In these systems, both PGF₂ and PGE₂ in hemolymph obtained from a single prawn were measured. PGs were extracted through reverse-phase C18 cartridge column, separated for isomers of each PG by high-performance liquid chromatography (HPLC), and quantified with a radioimmunoassay (RIA). Detectable range was 6.5–200 and 26–400 pg/tube for the PGF₂ and PGE₂ assays, respectively. Serial dilution of extracted and separated hemolymph PGs (RIA sample) produced a curve parallel to the standard curves in both assays. These assays were able to measure PGs in the RIA samples spiked with the authentic standard. Overall, results indicate that the presently established assay systems can accurately measure hemolymph PGs in female kuruma prawns.

When hemolymph PGF₂ and PGE₂ levels were determined and plotted against the gonado-somatic index (GSI), no correlation was found in either PGs. In terms of ovarian developmental stages, however, the concentration of PGF₂ significantly increased from 66.7 pg/ml at stage I (previtellogenic stage) to 303 pg/ml at stage II (primary vitellogenic stage) and decreased gradually thereafter. The concentration of PGE₂ also increased markedly (4037.7 pg/ml) at stage II and decreased at stage III (secondary vitellogenesis stage). These results indicate that PGs vary in concentration during ovarian development in kuruma prawns.     

Pharmacokinetics of oxolinic acid in black tiger shrimp, Penaeus monodon Fabricius, and the effect of cooking on residues

This study examined the pharmacokinetics and bioavailability of oxolinic acid (OA) in black tiger shrimp Penaeus monodon Fabricius, in brackish water (salinity 10 g L^?1) at 28–29°C, after intra‐sinus (10 mg kg^?1) and oral (50 mg kg^?1) administration and also investigated the net changes of OA residues in the shrimp after cooking (boiling, baking and frying). The haemolymph concentrations of OA after intra‐sinus dosing were best described by a two‐compartment open model. The distribution and elimination half‐lives were 0.84 and 17.7 h respectively. The apparent volume of distribution at a steady state and the total body clearance were estimated to be 2061 mL kg^?1 and 90.1 mL kg^?1 h^?1 respectively. The bioavailability of OA after an oral administration was 7.9%. The peak haemolymph concentration, the time to peak haemolymph concentration and the elimination half‐life after oral administration were 4.20 μg mL^?1, 4 h and 19.8 h respectively. Oxolinic acid muscle and shell levels increased to a maximum (muscle 1.76 μg g^?1 and shell 8.17 μg g^?1) at 4 h post administration and then decreased with the elimination half‐life value of 20.2 and 21.9 h respectively. Residual OA in muscle and shell was reduced by 20–30% by each cooking procedure examined.     

Efficacy, tissue distribution, and residue depletion of oxytetracycline in WS-RLP infected California red abalone Haliotis rufescens

Oxytetracycline (OTC) was recently demonstrated to be an effective therapeutant for California red abalone (Haliotis rufescens) infected with the agent of withering syndrome (WS), a Rickettsia-like prokaryote (WS-RLP). This study examined the influence of temperature on the efficacy and pharmacokinetics of oral OTC treatments on WS-RLP infected red abalone, H. rufescens. Medication was administered in an artificial diet containing 1.85% active OTC at a rate of 103.4 mg/kg abalone for 10, 20 and 30 days at both 13.4 °C and 17.3 °C. Drug residue concentrations, WS-RLP burden and WS-associated pathological changes within foot muscle and digestive gland were measured 3, 17, 23, 42, 63, 81, 102, 122, and 160 days after OTC treatment. In all samples the highest concentrations of OTC were detected on the first sample day. Furthermore, all sampling periods showed significantly less OTC in foot muscle samples relative to corresponding digestive gland samples (p < 0.001). While drug depuration occurred rapidly from foot muscle, unusually high concentrations and long depuration times were measured within the digestive gland. The pharmacokinetic trends in both the foot muscle and digestive gland at 13.4 °C were adequately described by a one-compartment model, while the digestive gland depuration models within the 17.3 °C treatments conformed to a two-compartment model. At both temperatures treatment duration gave rise to significant differences in OTC accumulation and depletion (p < 0.001). Despite this, terminal elimination rate constants were similar. Estimated half-lives were similar among all treatments for digestive gland (23.8–27.5 d) and all treatments for the foot muscle at 13.4 °C (10.7–13.4 d; calculated only for the 13.4 °C treatments). At both temperatures all three durations of drug treatment led to significant reductions in WS-RLP prevalence, foot muscle atrophy, and mortality. WS-RLP re-challenge trials initiated 44, 88, and 122 days following the 10-day treatment at 17.3 °C showed reduced susceptibility to re-infection for up to 88 days after treatment, suggesting that digestive gland OTC residues confer long-term resistance to pathogen. Atomic absorption spectroscopy conducted on both foot muscle and digestive gland found significantly higher concentrations of iron, zinc, and manganese in the digestive gland, offering an explanation behind the underlying mechanism of OTC retention for extended periods within this tissue.     

水产品中噁喹酸残留量的检测技术研究

采用高效液相色谱荧光检测器研究了牙鲆、罗非鱼和对虾肌肉组织中噁喹酸残留的检测技术。样品前处理方法为：以无水乙酸乙酯作为提取液,样品经均质、离心后,将提取液旋转蒸发浓缩,用流动相溶解定容,正己烷除去脂肪。高效液相色谱条件为：色谱柱采用反相C18柱,荧光检测器激发波长为325nm,发射波长为369nm,流动相为0．02mol／L磷酸＋乙腈＋四氢呋喃（69：16：15）,流速为0．8ml／min。噁喹酸标准曲线线性范围在10～200ng／ml。不同浓度水平下,噁喹酸的回收率为79％～93％,变异系数小于10％,检测限为10μg／kg。     

The effects of ozone and probiotics on the survival of black tiger shrimp (Penaeus monodon)

Total ozone production (TOP) from an ozonator, residual ozone concentration (ROC) in water, and the effects of ozone with or without probiotic supplemented feeds on bacterial growth, and shrimp (Penaeus monodon) survival were investigated. Minimal effective ROC to inhibit 3 log units of Vibrio harveyi D331 for 6 h and 2 log units of Bacillus S11 for 9 h was 0.38 g O₃/l of ROC from 5-min ozonation. Shrimp postlarvae exposed to 0.34–0.50 mg O₃/l ROC (8-h ozonation) caused loss of balance, immobility and destruction of gill lamellar epithelium. In vivo treatment of juvenile P. monodon reared on probiotic feed for 1 month revealed that 0.35 mg O₃/l ROC (30-min ozonation) effectively inhibited 3 log units of V. harveyi D331 for 24 h. At this ROC dosage, there was no effect either on shrimp or on intestinal probiotics (Bacillus S11). Shrimp survival from probiotic treatment, coupling with ozonation, increased significantly (P≤0.05) compared with controls.     

日本囊对虾卵母细胞发育过程中组织蛋白酶C比活性测定

组织蛋白酶C是一种溶酶样半胱氨酸蛋白酶,近年我们报道了组织蛋白酶C的表达与日本囊对虾(Marsupenaeus japonicus) 卵母细胞最后成熟阶段密切相关,本研究对卵母细胞发育主要不同时相特别是卵母细胞最后生理成熟过程进行了连续观察,测定了组织蛋白酶C在卵母细胞发育过程中的比活性。结果表明,日本囊对虾卵母细胞发育可划分为9个时相,即卵原细胞时相、核仁时相、周边核仁时相、初级卵黄发生时相、次级卵黄发生时相、皮质棒早期时相、皮质棒中期时相、皮质棒晚期时相、排卵期时相,生发泡的破裂是在排卵之前就已经发生;免疫印迹实验结果显示组织蛋白酶C蛋白只在卵母细胞最后成熟阶段即皮质棒发生时期表达,但酶活性测定结果表明组织蛋白酶C比活力在即皮质棒发生前后没有变化,酶活性较低,以上数据提示组织蛋白酶C可能与受精时皮质棒的释放以及受精卵外胶膜的形成有关。     

Structure and function of antimicrobial peptide penaeidin-5 from the black tiger shrimp Penaeus monodon

The gene for penaeidin-5, an antimicrobial peptide comprising 55 amino acids, was isolated from the hemocyte of black tiger shrimp (Penaeus monodon). RT-PCR expression tests revealed that penaeidin-5 was produced in hemocytes, gills, the intestine and muscle. Western blot analysis confirmed the panaeidin-5 was aboundantin hemocytes, the intestine and hemolymph. Immunohistochemistry revealedpenaeidin-5 in the cuticle and gills that are considered primary defense barriers. The deduced amino acid sequence of penaeidin-5 included a proline-rich N-terminal domain and a carboxyl-domain that contained six cysteine residues. Circular dichrosim analysis revealed an -helix in its secondary structure and the predicted 3D structure indicated two-disulfide bridges in the -helix. Based on the sequence of penaeidin-5 peptide cDNA, synthetic penaeidin-5 was prepared to carry out functional tests. The synthetic peptide had efficient bacteriostatic and bactericidal activity against Aerococcus viridans, and also inhibited the growth of two filamentous fungi, Fusarium pisi and Fusarium oxysporum. To measure penaeidin-5 in vivo, black tiger shrimp were challenged with Vibrio alginolyticus and A. viridans. At 3 h post-challenge, penaeidin-5 was induced and bacterial numbers decreased significantly by 12 h and 24 h.     

Pharmacokinetics and tissue distribution of oxytetracycline in sea bass, Dicentrarchus labrax, at two water temperatures

A pharmacokinetic study of oxytetracycline (OTC) following an intravascular administration (40 mg/kg) was carried out in sea bass, Dicentrarchus labrax (110 g), at 13.5 and 22 °C water temperature. Blood, muscle and liver samples were taken at 1, 2, 4, 8, 16, 32, 64 and 128 h post-injection. The plasma data were conformed to a two-compartment model. The kinetic profile of the drug was found to be temperature dependent. The absorption half-life (t_1/2) of OTC was 0.98 and 0.192 h at 13.5 and 22 °C, respectively, whereas the elimination half-time (t_1/2β) of the drug was 69 h at 13.5 °C and 9.65 h at 22 °C. The apparent volume of distribution of the drug at steady state [V_d(ss)] was 5.62 l/kg at 13.5 °C and 2.59 l/kg at 22 °C. The mean residence time (MRT) of OTC was found to be 37.7 h at 22 °C and 71 h at 13.5 °C. The total clearance of the drug (CL_T) was calculated to be 73.5 and 68.7 ml/kg/h at 13.5 and 22 °C, respectively.

Liver levels indicated higher OTC values than respective muscle levels at all time points and for both temperatures. The elimination of OTC from tissues tested was faster at the high temperature, whereas the drug was eliminated faster from liver compared to muscle when comparisons are made at the same temperature.     

Dopamine modulates the physiological response of the tiger shrimp Penaeus monodon

Levels of glucose, lactate, pO₂, pCO₂, HCO₃⁻, TCO₂, Na⁺, K⁺, Cl⁻, protein, and oxyhemocyanin in the hemolymph and its osmolality and pH were measured when tiger shrimp, Penaeus monodon (13.5 ± 1.5 g body weight), were individually injected with saline or dopamine at 10^− 8, 10^− 7, or 10^− 6 mol shrimp^− 1. Results showed that hemolymph glucose, lactate, pCO₂, HCO₃⁻, and TCO₂ values increased from 2 to 4 h; hemolymph osmolality, Na⁺, and total protein had increased at 2 h; and hemolymph K⁺ decreased from 2 to 8 h after the dopamine injection. All physiological parameters returned to the control values 4–16 h after receiving dopamine. The dopamine injection also significantly decreased the oxyhemocyanin/protein ratio of P. monodon which occurred at 2 h, resulting from an elevation of hemolymph protein and a slight decrease of oxyhemocyanin. These results suggest that stress-inducing dopamine caused a transient period of modulation of energy metabolism, osmoregulation, respiration, and the acid–base balance in P. monodon in adapting to this environmental stress.     

噁喹酸在欧洲鳗鲡体内的药代动力学及残留研究

在(25±1)℃水温条件下,分别采用口灌和浸浴的给药方式,以20 mg/kg的单剂量混饲口灌及5 mg/L的剂量浸浴18 h给予噁喹酸后,用高效液相色谱法测定血浆和肌肉中的药物浓度,研究了不同给药方式下噁喹酸在欧洲鳗鲡(Anguilla anguilla)体内的药代动力学特征.结果表明:欧洲鳗鲡口灌给药后,其血浆和肌肉...     

A new and practical application of JH antagonist KK-42 to promoting growth of shrimp Penaeus schmitti

The shrimps (Penaeus schmitti), 0.6–0.8 cm long, were soaked for 1 min in juvenile hormone antagonist KK-42 solution at a concentration of 1.95 × 10^− 4 mol/L, and then cultured in normal way. The body weight of animals rose with increasing culture time in all groups, but was higher in KK-42 treatment. The increase rate of body weight of treatment group was 167% and 68% higher than control group at 30th day and 60th day, respectively; the body weight rose by 42.5% at 90th day and 22.1% at 120th day. Histological observations showed that the number of acini of mandibular organ increased gradually as animals grew, and the diameter of acinus-forming cells was statistically bigger at 120th day than at other times. After KK-42 treatment, both the number of acini and the diameter of gland cells decreased in the entire experiment observed. The results indicated the inhibition of anti-juvenile hormone KK-42 on the development of mandibular organ in P. schmitti.     

Pharmacokinetics of oxolinic acid in gilthead sea bream, Sparus aurata L.

This is the first study on the pharmacokinetic parameters of oxolinic acid (OA) in gilthead sea bream, Sparus aurata L. The kinetic profile of OA was studied after a single intravascular injection (20 mg kg⁻¹) in 100 g fish at 20 °C. The distribution half-life ( t _1/2α) and the elimination half-life ( t _1/2β) of the drug were found to be short (0.51 and 12.60 h, respectively). The drug penetration from the plasma to the tissues was adequate as the apparent volume of distribution of the drug at steady-state ( V _d(ss)) was found to be 2.11 L kg⁻¹. The mean residence time ( MRT ) of OA was short (14.25 h) and the total clearance rate ( Cl _T) of the drug was low (0.15 L kg⁻¹ h⁻¹). The bioavailability ( F %) of OA following oral administration (30 mg kg⁻¹) was also low (14%). Maximum values were observed for muscle at 0.5 h after injection, with levels declining as with subsequent sampling. At the first two time points (0.5 and 1 h) plasma levels of OA were higher than muscle, however, the reverse was evident for subsequent samples. Following oral administration, highest muscle levels were found at 16 h and, with the exception of the 24-h sampling, muscle OA concentrations were higher than plasma at all time points. The fast elimination of OA suggests short withdrawal times with reference to human consumption of treated fish.     

基于线粒体Cytb基因探讨我国日本囊对虾4个地理群体的遗传结构及种群分化

为了分析日本囊对虾两种表型差异类型的系统发育关系,实验系统比较了两种类型线粒体全基因组的基因结构、遗传距离和密码子偏好性等特征。结果显示,两种类型线粒体基因组的基因重叠区、基因间隔和密码子等方面均存在差异。两种类型的线粒体蛋白编码基因核苷酸序列的相似度为91.10%~95.00%,氨基酸序列的相似度为96.15%~100%。两种类型的A+T富集区的同源性只有82.60%,分歧度为15.40%,呈现高度遗传分化。日本囊对虾两种类型蛋白编码基因的核苷酸序列分歧度大于明对虾属和叉肢螯虾属,但氨基酸序列的分歧度小于后者。研究发现,基于cytb基因的种间遗传距离大于种内遗传距离的10倍;基于cox1基因的种间遗传距离分别为类型Ⅰ和类型Ⅱ的种内遗传距离的14.6倍和5.2倍。两种类型的nd1、nd4和nd5基因的Ka/Ks值大于1,表明受到正向选择。基于20种隶属9属的对虾线粒体基因组蛋白编码序列构建系统发育树,结果显示能有效区分各属对虾,其中日本囊对虾的两种类型首先聚类,再与宽沟对虾聚类。研究表明,日本囊对虾的两种表型差异类型基本达到物种水平,有必要进一步评估更多的性状差异,以提高两种囊对虾的特异性养殖技术。     

斑节对虾幼虾不同家系的耐低盐能力比较

为了选育斑节对虾耐低盐新品系,以11个不同品系亲本的斑节对虾家系作为材料,使用0、0.5以及1三个盐度对其进行急性胁迫实验。结果显示,盐度1对斑节对虾的存活率影响不大,各家系都能良好适应,所以不适合作为评估家系耐低盐能力的胁迫浓度。盐度0.5与盐度0对斑节对虾的胁迫作用较强,各家系最终全部死亡,半致死时间的存活率在各家系间存在显著差异,而且盐度0与盐度0.5胁迫的结果具有一定一致性,所以盐度0与盐度0.5都可以作为评估斑节对虾家系耐低盐能力的胁迫浓度。研究表明,J(♀61619×♂61548非×泰)、I(♀61173×♂61615泰×非)、H(♀61116×♂61625泰×非)和K(♀61689×♂61504非×泰)4个家系的耐低盐能力较强,可以作为候选家系应用于耐低盐家系的选育工作。     

运用脂肪酸标志法分析刺参食物来源的季节变化

运用脂肪酸标志法分析了刺参的食物组成及季节变化。刺参饵料中含有硅藻、褐藻、多种异养细菌、大型绿藻、鞭毛藻或原生动物等,其中硅藻、褐藻和细菌在全年的食物贡献比较大,各种饵料来源比例具有显著的季节变化。实验期间,刺参体壁的硅藻脂肪酸标志22:5(n-3)相对含量很高(7.24%～14.45%),且16:1(n-7)/16:0比值全年在0.73～1.82之间(平均1.10),表现出典型的硅藻脂肪酸特征,表明硅藻是刺参主要的食物来源。褐藻脂肪酸标志20:4(n-6)在刺参体壁脂肪酸组成中相对含量较高(4.88%～8.16%),且在秋冬季节达到较高水平,表明秋冬季节褐藻类对刺参的食物贡献可能较大。噬纤维菌—黄杆菌类的脂肪酸标志奇数碳及支链脂肪酸(Odd&brFAs,5.31%～8.29%)和变形细菌的脂肪酸标志[18:1(n-7),5.85%～6.86%]相对含量比较高,表明细菌在全年都是刺参重要的食物来源。主成分分析发现,1月份刺参的主要食物来源是硅藻、鞭毛藻或原生动物、褐藻及细菌;3月份硅藻、鞭毛藻或原生动物、大型绿藻的食物贡献较大;6月份大型绿藻在刺参的食物来源中占较大比重;7月份细菌和大型绿藻的食物贡献较大,细...