Effect of parental origin on early life history traits of European eel

Establishment of European eel (Anguilla anguilla) hatchery production will rely on selectively bred individuals that produce progeny with the best traits in successive generations. As such, this study used a quantitative genetic breeding design, between four females and nine males (four wild‐caught and five cultured), to investigate the effect of paternal origin (wild‐caught vs. cultured) and quantify the relative importance of parental effects, including genetic compatibility, on early life history (ELH) performance traits (i.e. fertilization success, embryonic survival at 32 hr post‐fertilization, hatch success and larval deformities at 2 days post‐hatch) of European eel. Wild‐caught males had higher (56%) spermatocrit values than cultured males (45%), while fertilization success, embryonic survival, hatch success and larval deformities were not significantly impacted by paternal origin. This demonstrates that short‐term domestication of male eels does not negatively affect offspring quality and enables the consideration of cultured male broodstock in future breeding programmes. Moreover, paternity significantly explained 9.5% of the variability in embryonic survival, providing further evidence that paternal effects need to be taken into consideration in assisted reproduction protocols. Furthermore, maternity significantly explained 54.8% of the variation for fertilization success, 61.7% for embryonic survival, 88.1% for hatching success and 62.8% for larval deformities, validating that maternity is a major factor influencing these “critical” ELH traits. At last, the parental interaction explained 12.8% of the variation for fertilization success, 8.3% for embryonic survival, 4.5% for hatch success and 20.5% for larval deformities. Thus, we conclude that eggs of one female can develop more successfully when crossed with a compatible male, highlighting the importance of mate choice for successful propagation of high‐quality offspring. Together, this knowledge will improve early offspring performance, leading to future breeding programmes for this critically endangered and economically important species.     

超低温冷冻对日本鳗鲡精子酶活性的影响

研究超低温冷冻保存(-196℃)对日本鳗鲡精子内总ATP酶、肌酸激酶(CK)、琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽还原酶(GR)等酶活性的影响。运用试剂盒分别测定了冷冻前后日本鳗鲡精子内酶活性的变化。结果表明,经过超低温冷冻保存后,日本鳗鲡精子的活力下降,精子内GR活性显著升高(P<0.05),酶活性从冻前的358.52±45.65 U/L上升到646.30±70.30 U/L;其它几种酶的活性均显著下降(P<0.05),总ATP酶、CK和SDH的活性分别从冻前的3.14±0.61 U/ml、17.10±3.51 U/ml和32±5.94 U/ml下降到1.83±0.43 U/ml、7.33±1.74 U/ml和21±1.41 U/ml,LDH、SOD和CAT活性分别从冻前的2 266.67±313.25 U/L、220.47±32.94 U/ml和48.51±5.94U/ml下降到1 195.91±198.51 U/L、84.16±22.11 U/ml和21.8±4.14 U/ml。超低温冷冻对日本鳗鲡精子酶活性和精子活力均有较大影响。     

鳗鲡肠道中嗜水气单胞菌的拮抗菌筛选

从鳗鲡肠道中分离到643株细菌,用点种法筛选出65株病原性嗜水气单胞菌的拮抗菌,研究拮抗效果最好的14株拮抗菌的抗菌谱、生长曲线以及与病原性嗜水气单胞菌的共凝集作用.研究结果显示：NAC琼脂培养基筛选的2株拮抗菌（Na3-38和Nb3-15）对7株指示菌均有较强的拮抗作用,Aa3-22和Mb3-7对7株指示菌中的5株有拮抗效果;除Na3-38和Nb3-15以外,其余12株的生长系数均大于嗜水气单胞菌;11株拮抗菌（Na3-38、Nb3-15、Ab2-55、Ab2-77、Ab1-3、Aa3-22、Aa3-67、Mb1-4、Mb2-10、Mb3-7和Mb3-27）和嗜水气单胞菌的共凝集率大于10%.这表明：Aa3-22和Mb3-7在抗菌谱、生长曲线以及与病原性嗜水气单胞菌的共凝集作用都适合作为益生菌.     

日本鳗鲡消化道肥大细胞组织化学的研究

研究日本鳗鲡（Anguilla Japonica）消化道组织中的肥大细胞组织化学性质,观察肥大细胞胞浆颗粒的异质性。采用改良甲苯胺蓝（MTB）、阿利新蓝-沙黄O（AB／SO）、甲基绿-派洛宁（MG—P）、和天青Ⅱ-伊红-瑞氏混合液等4种组化染色法。用阿利新蓝-沙黄O染色法显示日本鳗鲡消化道的肥大细胞效果较好;甲基绿-派洛宁（MG—P）、和天青Ⅱ-伊红-瑞氏混合液染色效果一般,被染的肥大细胞数量很少;改良甲苯胺蓝甚至观察不到肥大细胞。日本鳗鲡肥大细胞数量不多,主要分布于食管、胃和肠道等部位黏膜层的黏膜上皮下方、固有层结缔组织,少量分布在黏膜下层结缔组织,并且有沿血管周围分布的特点;肥大细胞体积小,且多呈圆形、椭圆形、梭形及不规则形。AB／SO染色法效果最好,适用于日本鳗鲡肥大细胞的鉴定;改良甲苯胺蓝染色法不适合于日本鳗鲡肥大细胞的鉴定。     

注射含HCG和CPE的W／O／W复乳诱导日本鳗鲡性腺发育成熟的研究

用含HCG及鲤脑垂体（PG）匀浆液提取物（CPE）的W／O／W复乳，对塘养雌鳗进行注射催熟实验，雌鳗平均GSI达35．18％，最高达56．5％，其催熟效果较常规注射方法（平均GSI 27．9％，最高50．4％）好，用含LHRH－A，HCG，CPE的复乳和DOM生理盐水混悬剂对部分达到性成熟的鳗鲡进行催产，用W／O／W复乳诱导成熟的鳗鲡，排卵率为75％，用常规方法催熟的鳗鲡排卵率为55％，注射复乳制剂后，鳗鲡血清GtH含量可较长时间的保持较高水平，血清GtH的变化较常规注射方法缓和，注射乳剂还可使鳗鲡脑垂体GtH含量升高。     

人工催熟日本鳗鲡精子的显微和超微结构

对经人工催产获得的日本鳗鲡（Anguilla japonica）精子的显微和超微结构进行了观察。光镜下观察,日本鳗鲡精子有两种形态结构,一种精子的细胞核为圆形或近 似圆形,这精子较小,细胞核长径为(2.57±0.62) μm,短径为(2.11±0.59) μm,鞭毛长度为(38.35±7.71) μm;另外一种精子的细胞核为“眉形”或“新月形”,精子较大,细胞核长径为(7.66±1.09) μm,短径为(2.54±0.46) μm,鞭毛长度为(38.35±9.02) μm。透射电镜观察结果显示：圆形精子头部的顶端无顶体,植入窝位于细胞核底端的中间,由细胞核向内凹陷而成,呈一沟状,其走向与精子的长轴平行。中心粒复合体位于植入窝内。细胞核的下端有2～3个线粒体。基体的头端呈筒状,由电子致密物构成。基体的尾端分裂成9束。尾部从袖套腔中伸出且细长,轴丝始端与基体的尾端相连,微管多呈电子致密状,轴丝为典型的“9+2”结构。圆形精子为日本鳗鲡正常成熟的精子。“眉形”或“新月形”精子头部细胞核弯曲,在弯曲处有一圆形的球状物,球状物内含有线粒体和中心粒复合体,轴丝为“9+0”结构。这种精子可能存在发育缺陷或没有达到完全正常成熟。     

欧洲鳗鲡血清免疫球蛋白纯化及其结构分析

应用亲和层析技术提纯欧洲鳗鲡免疫球蛋白（Ig）,并对欧鳗Ig的结构和抗原性进行分析。层析结果表明：Ig蛋白呈现一个锐形曲线,蛋白峰与抗原活性峰高度重叠。SDSPAGE分析表明：欧鳗Ig重链分子量为68 kD,轻链有3条,分子量分别为21 kD、23 kD和26 kD。凝胶电泳结果显示：在非变性非还原条件下,欧鳗Ig有790 kD和350 kD 2条蛋白带,在变性非还原条件下有790 kD、593 kD和350 kD 3条蛋白带。Western-blotting试验证实：兔抗欧鳗Ig能识别欧鳗Ig的多种不同聚合体和Ig的重链,但不能识别Ig的轻链。结论：欧鳗Ig在自然条件下可能以四聚体和二聚体的形式存在,这与其它硬骨鱼的Ig形式有差异。欧鳗Ig链间二硫键不健全,在SDS作用下可解聚产生多种不同分子量的聚合体,首次揭示欧鳗Ig的轻链有3种异型。     

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.     

欧洲鳗鲡三类苗养殖技术初探

本试验应用膨化饲料养殖欧洲鳗鲡（Anguilla anguilla）三类苗。经过110d的养殖,两个试验组的平均体重分别达到16．13g和15．39g,相对增重率分别为151．1％和142．4％。日均增重量分别为88．3mg和82．2mg。平行养殖试验结果显示存活率分别为97．7％和98．5％,饲料转化率分别为59．1％和58．4％。整个养殖过程基本不换水,完全不用任何药物。