Sperm production and quality in European eel (Anguilla anguilla) in relation to hormonal treatment

Aquaculture production relies on controlled management of gametogenesis, especially in species where assisted reproduction is needed for obtaining gametes in captivity. The present study used human chorionic gonadotropin (hCG) treatments to induce and sustain spermatogenesis in European eel (Anguilla anguilla). The aim was to evaluate effects of strip-spawning timing (12 vs. 24 hr) after weekly administration of hCG and the necessity of a primer dose (in addition to weekly hormonal treatment) prior to strip spawning (primer vs. no-primer) on sperm quality parameters. Sperm parameters included milt production (weight), density and sperm kinematics at Week 9, 11 and 13 after onset of treatment. Spermiation commenced in 11.5% of males in Week 5 and by Week 9, and all males produced milt. Male weight, milt production, sperm density and spermatocrit did not differ among hormonal treatments during the experimental period. Overall, male weight decreased from 106.3 to 93.0 g, milt weight increased from 3.5 to 5.4 g, sperm density counts decreased from 11.7 × 10⁹ to 10.5 × 10⁹ cells/ml, and spermatocrit decreased from 46.5% to 40.5%. Furthermore, spermatocrit was positively related to haemocytometer counts (R² = .86, p < .001), providing a reliable indicator of sperm density. Differences in sperm kinematics were observed depending on strip-spawning timing after hormonal injection (12 vs. 24 hr) but with no consistent pattern. These sperm quality parameters also did not consistently differ between the no-primer and primer treatments. Considering that each male may be stripped 4–5 times over the 2–3 months spawning season, omitting the primer would reduce animal handling, material costs and labour intensity, while sustaining high-quality sperm production.     

Molecular cloning and characterization analysis of immunoglobulin M heavy chain gene in European eel (Anguilla anguilla)

In this study, the immunoglobulin M heavy chain gene of European eel (Anguilla anguilla) was cloned and analyzed. The full-length cDNA of the IgM heavy chain gene (GenBank accession no. EF062515) has 2089 nucleotides encoding a putative protein of 581 amino acids. The IgM heavy chain was composed of leader peptide (L), variable domain (VH), CH1, CH2, Hinge, CH3, CH4, and C-terminus and two novel continuous putative N-glycosylation sites were found close to the second cysteine of CH3 in A. anguilla-H1 and A. anguilla-H2. The deduced amino acid sequence of the European eel IgM heavy chain constant region shared similarities to that of the Ladyfish (Elops saurus), Atlantic salmon (Salmo salar), rainbow trout (Oncorhynchus mykiss), Grass carp (Ctenopharingodon idella), Common carp (Cyprinus carpio), Channel catfish (Ictalurus punctatus), and the orange-spotted grouper (Epinephelus coioides) with the identity of 46.1%, 39.7%, 38.9%, 32.4%, 32.3%, 31.7%, and 30.7%, respectively. The highest level of IgM gene expression was observed in the kidney, followed by the spleen, gills, liver, muscle and heart in the apparently healthy European eels.     

饲料中添加糖代谢调节因子对欧洲鳗鲡生长的影响

通过在欧洲鳗鲡(Anguilla anguilla L.)幼鱼饲料中添加糖代谢调节因子(喜福得),在室内循环水族箱中饲养初始体重为41.07±0.37g的欧洲鳗鲡56d,研究喜福得对其生长性能的影响。结果表明,饲料中添加0.1%的喜福得能促进欧洲鳗鲡的生长,试验组饲料的幼鳗增重率、饲料效率及蛋白效率与对照组相比分别提高了18.4%、11.2%和11.48%。     

Pharmacokinetic and depletion studies of sarafloxacin after oral administration to eel (Anguilla anguilla).

The pharmacokinetics of sarafloxacin applied by oral gavage at a dose of 15 mg/kg b.w. was studied in eel (Anguilla anguilla) at water temperature of 24 degrees C. Sarafloxacin levels were determined using high performance liquid chromatography with a quantitation limit of 0.07 microg/ml or gram. The time to peak plasma concentration, Tmax, was 12 hr and peak concentration, Cmax, was 2.64 microg/ml. The absorption rate constant (k(a)) was 0.23 hr(-1) (r=0.996). The drug disposition curve after Tmax was fitted to a two-compartment open model. The distribution rate constant (alpha) was 0.085 hr(-1) (r=0.972), and the half-life (t(1,2alpha)) was 8.15 hr. The elimination rate constant (beta) was 0.023 hr(-1) (r=0.909), and the half-life (t(1/2beta)) was 30.13 hr. The estimated area under the curve, AUC, was 56.7 microg.hr/ml. The peak concentrations of drug in liver, kidney, muscle, and skin were 13.39 (12 hr), 5.53 (12 hr), 1.82 (24 hr), and 0.78 microg/g (40 hr), respectively. The time for sarafloxacin mean levels to fall below detectable limits in the plasma, muscle, and skin were 7 days but for the liver and kidney were 14 days.     

Single-dose pharmacokinetics of flumequine in the eel (Anguilla anguilla) after intravascular, oral and bath administration

Knowledge of the pharmacokinetic properties of drugs to combat bacterial infections in the European eel (Anguilla anguilla) is limited. One antimicrobial agent likely to be effective is flumequine. The aim of this study was to investigate the pharmacokinetic properties of flumequine in European eels in fresh water. Flumequine was administered to eels (Anguilla anguilla) intravenously (i.v.) and orally (p.o.) at a dose of 10 mg/kg body weight, and as a bath treatment at a dose of 10 mg/L water for 2 h. The study was performed in fresh water with a temperature of 23 + 0.3 degrees C, pH 7.15. Identical experimental designs were used. Two additional bath treatments were also performed, one in which the pH in the water was lowered by approximately 1 unit to 6.07 (dose: 10 mg/L) and one at a dose of 40 mg/L for 2 h in a full-scale treatment. Following i.v. administration, the volume of distribution at steady state was 3.4 L/kg. Total body clearance was 0.012 L/h per kg and the elimination half-life (t1/2lambda z) was calculated to be 314 h. Mean residence time was 283 h. Following oral administration, the t1/2lambda z was 208 h. Maximal plasma concentration (Cmax) was 9.3 mg/L, at 7 h after administration (Cmax). The oral bioavailability (F) was calculated to be 85%. Following bath administration in 10 mg/L for 2 h, maximal plasma concentration was 2.1 mg/L, observed immediately after the end of the bath. The 'bioavailability' in eel following a 2-h bath treatment was 19.8%. Reducing the pH in the bath to 6.07 produced a maximal plasma concentration of 5.5 mg/L, observed immediately after the end of the bath. The 'bioavailability' was increased to 41% by the lowering of the pH. A similar effect was observed in a full-scale treatment (1 kg eels/L water). The CO2 produced by the eel lowered the pH and increased 'bioavailability' to 35%.     

动物精液品质评定方法的研究进展

人工授精和体外受精技术的广泛应用,使精液保存技术得到了快速发展。精子的质量和功能状态直接影响其受精能力。文章综合阐述了近几年动物精液品质评定的方法:精子质膜完整性检测、线粒体活性的测定、受精能力的检测和精液中酶活性的测定。     

欧鳗(Anguilla anguilla)“狂游症”病调查报告

本文在国内首次较系统报告了欧鳗“狂游症”病的流行病学、病理组织学、诊断和防治方法，同时在国内外首次报告了侵袭欧鳗脑部的病毒是该病的致病原，这对今后深入研究起到很大的作用。     

动物精液质量检测方法研究进展

家畜精液质量的检测对于人工授精技术和体外受精技术的应用具有重要意义。目前,在人类生殖临床和畜牧生产中,评定精液质量的方法大多是几个指标联合检测,使得精液质量评价的结果更加科学和准确。文章综述了近年来家畜精液质量检测的新技术、新方法和新进展,以便较确切的反映精子的质量与受精能力。     

新型欧洲鳗鲡抗菌肽elecilin在毕赤酵母中的表达

前期采用差减杂交的方法筛选到新型欧洲鳗鲡抗菌肽elecilin,其基因全长为477 bp,编码159个氨基酸,理论分子量约17.4 kDa。为了获得具活性的大量表达的抗菌肽,本研究利用毕赤酵母表达了elecilin。首先将elecilin基因克隆至酵母分泌表达载体pPICZaA,构建了重组分泌表达质粒pPICZaA-elecilin,电击转化毕赤酵母X-33。经Zeocin筛选和PCR鉴定,获得重组酵母菌,通过甲醇诱导,SDS-PAGE和Western blot鉴定,证实诱导后的培养基中存在与预期分子量大小相一致的蛋白,能被His-tag单克隆抗体识别,表明利用酵母成功表达了欧洲鳗鲡抗菌肽elecilin。这为进一步研究该抗菌肽的功能及作为饲料添加剂的应用奠定了基础。     

The in vitro effect of bovine lactoferrin on the activity of organ leukocytes in rainbow trout (Oncorhynchus mykiss), European eel (Anguilla anguilla) and wels catfish (Silurus glanis)

Lactoferrin (LF) is a glycoprotein found in milk, neutrophil granules, secretions and selected organs of mammals. Lactoferrin exhibits antibacterial, antiviral, fungicidal, immunoregulatory and other functions. Although fish are devoid of this protein and its cell receptors, LF effect on the immune mechanisms of fish has been demonstrated. The objective of this study was to investigate the effect of bovine lactoferrin, applied in vitro, on the activity of head kidney and spleen leukocytes in three freshwater fish species: rainbow trout (Oncorhynchus mykiss), European eel (Anguilla anguilla) and wels catfish (Silurus glanis). The obtained results validate LF beneficial effect on the respiratory burst of phagocytes in rainbow trout and wels catfish despite the fact that the potential killing activity against Aeromonas hydrophila was not stimulated in any of the studied species. Bovine lactoferrin enhanced the proliferation of T-lymphocytes in rainbow trout and European eel, as well as of B-lymphocytes in rainbow trout.     

四种活体染色法评定家禽精子形态的研究

精子形态学检测是预测精子功能最有价值的指标之一,形态异常精子越多受精率越低。用4种伊红-苯胺黑活体染色法(Blom、Bakst、Morisson、Jaskowski)评价三黄鸡和贵妃鸡的精子形态,结果表明:染色方法显著影响精子形态,但不影响精子活率,染液对精子自然形态的保持至关重要。Blom染色法导致精子头部膨胀,畸形精子显著增多(P≤0.01),Bakst、Morisson和Jaskowski染色法均可作为评定家禽精子形态的实用方法,其中Jaskowski染色方法最简便经济,值得在家禽生产中推广应用。     

微生态制剂在欧鳗饲料中的应用

福建省福州开发区高龙饲料有限公司在福建三明明溪县华盛特种养殖场进行添加微生态制剂试验 ,微生态制剂选用益菌多 (由北京桑普提供 ) ,益菌多主要含有乳酸杆菌 ,粪链球菌和芽孢杆菌 ,它们是动物消化道正常微生物菌群 ,活菌数≥ 10 9ｃｆｕ/ｇ。1　材料与方法1.1　材料与条件试验池为正方形 ,水泥砖混结构 ,上方履盖遮阳网的增氧池 ,养殖品种为欧洲鳗鲡 ,用水是山溪水 ,水温为 2 4～ 2 6℃ ,一组为对照组 ,另一组为试验组 ,对照组用高龙幼鳗饲料 ,幼鳗饲料营养指标见表 1;试验组在对照组饲料中额外补加 0 .2 %的益菌多。试验从 2 0 0 2年 7…     

Effect of Sperm Cryopreservation on the European Eel Sperm Viability and Spermatozoa Morphology

The main objective of the present work was to study the effect of cryopreservation of European eel sperm both on the sperm viability and the spermatozoa head morphology. Spermatozoa morphology was evaluated with computer-assisted morphology analysis after collection in fresh samples, after adding the freezing medium containing dimethyl sulfoxide as cryoprotectant and, finally, after the cryopreservation process and thawing. Cell viability was assessed, in both fresh and thawed samples, by Hoechst 33258 staining. Computer-assisted sperm analysis (CASA) was used to determine the percentage of motile cells and to measure motility parameters in sperm samples. A significant decrease of head perimeter (12.56%) and area (17.90%) was detected from spermatozoa in fresh to thawed samples, indicating that cells do not recover the original size after the cryopreservation process. CASA was used to measure the percentage of motile cells (51.9%) and spermatozoa motility parameters such as curvilinear, straight line and angular path velocities, as well as beating cross frequency. This technique was employed in the fresh sperm samples but proteins present at the freezing medium (L-alpha-phosphatidylcholine) made impossible to use this last technique in thawed samples. When sperm viability was assessed by Hoechst staining, a significant decrease of approximately 15% (73.10 vs 58.26%) of alive spermatozoa was registered from fresh to thawed samples. The percentage of motile cells measured by CASA in fresh samples (51.9%) was lower than the percentage of alive cells determined by Hoechst stainning, suggesting the existence of different batches of spermatozoa in different stages of development, even during the eight to tenth weeks of treatment, when the highest sperm quality was found.     

欧洲鳗对几种蛋白质饲料的休外消化研究

利用体外消化法研究了欧洲鳗粗酶液对鱼粉、发酵豆粕、菜籽粕、玉米蛋白粉、啤酒酵母、蝇蛆粉、膨化大豆、豆粕、花生粕、红虫粉等10种单一蛋白源以及另外5种复合蛋白源酶解7 h的体外消化率以及酶解液中游离氨基酸的生成量,以期筛选出最有可能替代鱼粉的蛋白原料.结果表明:(1)最有望部分或全部取代鱼粉的植物性蛋白源为啤酒酵母、发酵豆粕和花生粕,其干物质和粗蛋白质体外消化率分别为62.75%、55.15%、49.45%和78.50%、70.70%、85.70%;动物性蛋白源中红虫粉和蝇蛆粉的干物质体外消化率与鱼粉差别不大,粗蛋白质消化率红虫粉＞蝇蛆粉＞鱼粉.(2)在0～7h,各种蛋白源酶解氨基酸生成量基本呈增加趋势,对植物性蛋白源来说,啤酒酵母氨基酸生成量明显高于其他植物性蛋白源,其次是发酵豆粕、玉米蛋白粉和花生粕,较低的为菜籽粕、膨化大豆、豆粕.对于动物性蛋白源来说,红虫粉体外消化生成氨基酸的量明显高于鱼粉和蝇蛆粉.这一结果说明红虫粉和啤酒酵母是优质的蛋白源,很容易被鳗鱼消化吸收利用.(3)对于混合蛋白源混1、混2、混3、混4、混5来说,其干物质和粗蛋白质的体外消化率分别为:混1＞混2＞混5＞混4＞混3;混2＞混1＞混5＞混4＞混3,而原料中粗蛋白含量为混3＞混4＞混5,其鱼粉添加量分别为80%、70%、60%,混1、混2均不含鱼粉,这说明鳗鱼对几种植物性蛋白源的粗蛋白体外消化率均高于鱼粉,这也与之前单一蛋白源的粗蛋白体外消化率结果一致.     

几种草食动物日粮植物组成研究技术和方法的比较

目前，用于评价草食动物日粮的植物组成的方法大致可分为７种：直接观察法、牧草食物法、瘤胃内容物法、瘘管法、粪便分析法、显微技术和红外及近红外反射光谱技术。     

常用哺乳动物精子质量检测方法

根据精子的特性 ,利用光学显微镜、荧光显微镜、流式细胞记数仪等仪器 ,结合常规染色技术或荧光探针技术 ,通过检测精子染色质的状态、运动能力、质膜的完整性、顶体的状态、线粒体的活性、获能、顶体反应以及与卵子的结合能力等指标来评价精子的功能状态 ,以便准确预测精子的受精能力。