近江牡蛎和长牡蛎的配子兼容性及合子育性

为了评估长牡蛎(GG)和近江牡蛎(AA)的配子兼容性和合子育性，本实验探究了盐度(16、20、24、28和32)、温度(18、21、24、27和30 ℃)和精子浓度(10⁰、10¹、10²、10³和10⁴个/μL)对受精率、胚胎畸形率和孵化率的影响。结果显示，随盐度的增加，GG组的受精率和孵化率逐渐上升，而AA组和AG组的受精率和孵化率先上升后下降；GA组和AG组分别在盐度28和24时具有较优的配子兼容性和合子育性，受精率、畸形率、孵化率分别为7.45%±5.05%、2.00%±0.90%、63.60%±9.88%和14.26%±9.26%、1.74%±0.93%、66.16%±9.43%。随温度上升，AA、GA和GG组的受精率和孵化率先升高后下降，而畸形率先下降后上升；GA组和AG组均在27 ℃时表现出较优的配子兼容性和合子育性，此时GA组和AG组的受精率、畸形率、孵化率分别为7.95%±4.04%、1.79%±1.04%、57.11%±9.95%和14.70%±7.27%、1.66%±0.85%、67.25%±15.19%。随精子浓度的增加，GG组的受精率逐渐上升，孵化率逐渐下降，GA组和AG组的受精率逐渐上升；GA组和AG组均在10⁴个/μL时表现出较优的配子兼容性和合子育性，此时二者的受精率、畸形率、孵化率分别为9.09%±7.53%、1.59%±0.48%、67.97%±19.96%和14.30%±6.04%、1.06%±0.68%、67.33%±12.65%。研究表明，温度、盐度和精子浓度对长牡蛎和近江牡蛎种间配子兼容性影响较小，但对合子育性影响较大。     

香港巨牡蛎与近江牡蛎杂交及回交子代早期生长发育比较

为评估香港巨牡蛎与近江牡蛎杂交及回交育种的可行性,本文以香港巨牡蛎(Crassostrea hongkongensis)与近江牡蛎(Crassostrea ariakensis)为材料,比较了两种牡蛎杂交与回交子代的受精率、孵化率、幼虫及稚贝的生长和存活率。结果表明：香港巨牡蛎卵子可以与近江牡蛎精子受精,受精率为47.33±2.52%,但近江牡蛎卵子与香港巨牡蛎精子不受精。在回交实验中,香港巨牡蛎(H♀)与近江牡蛎(A♂)的杂交牡蛎 (HA)与香港巨牡蛎(HH)或近江牡蛎(AA)都可以正常交配,但各回交组受精率明显低于两牡蛎自交组(P＜0.05)。受精后,HA与AA回交组的孵化率最低,分别为20.33±2.08%(♀A×♂HA),55.67±2.52%(♀HA×♂A),其它各实验组孵化率均大于70%。幼虫期, HA幼虫存活率明显低于两牡蛎自交组HH和AA(P＜0.05)；HA幼虫生长快于AA, 但慢于HH；♀A×♂HA回交组和HA自繁组的幼虫存活率为0。稚贝期,HA稚贝存活率与HH、AA自交组存活率差异不显著(P>0.05)。HA稚贝生长快于HH, 但慢于AA。另外,研究发现♀H×♂HA回交组幼虫和稚贝的生长与HA差异不显著(P>0.05),优于其它回交组；♀H×♂HA回交组幼虫的存活率与HA差异不显著(P>0.05),其稚贝的存活率虽低于HA,但优于其它回交组。本研究为牡蛎种间杂交及回交育种奠定了一定的理论基础。     

温度对长牡蛎和葡萄牙牡蛎及其杂交稚贝生长和存活的影响

为比较不同温度下长牡蛎(Crassostrea gigas)和葡萄牙牡蛎(C. angulata)杂交稚贝的生长与存活差异, 以正交组GA(长牡蛎♀×葡萄牙牡蛎♂)、反交组AG(葡萄牙牡蛎♀×长牡蛎♂)、自交组GG(长牡蛎♀×长牡蛎♂)和AA(葡萄牙牡蛎♀×葡萄牙牡蛎♂)的稚贝为材料, 开展了温度适应性研究。结果表明, GA和AG的亚致死温度为37.50 ℃, GG和AA的亚致死温度为37.00 ℃; GA致死温度为44.00 ℃, AG、GG和AA的致死温度为43.50 ℃; LT₅₀由高到低依次为GA (42.47 ℃)、AG (41.94 ℃)、AA (41.63 ℃)、GG (41.55 ℃)。多重比较表明, GA和AG的最适生存温度为16~28 ℃, GG为20~28 ℃, AA为24~28 ℃; GA、AG和GG的最适生长温度为20~28 ℃, AA为24~28 ℃。所有温度下杂交稚贝均表现出生长与存活优势。其中, 生长中亲杂种优势H和单亲优势H_AG在20 ℃时最高, 分别为35.1和38.8, 单亲优势H_GA在32 ℃最高, 为43.4; 16 ℃时存活中亲杂种优势H为28.4, 单亲优势H_GA和H_AG分别为21.5和36.4。32 ℃时中亲杂种优势H为42.2, 单亲优势H_GA和H_AG分别为72.7和20.0。研究表明, 杂交稚贝耐热性大于双亲, 具有显著的生长和存活优势, GA的优势更加明显。     

PI/Rh123双染和流式细胞术作用下不同盐度对葡萄牙牡蛎精子质量及受精的影响

以葡萄牙牡蛎精子为对象,采用碘化丙啶(PI)/罗丹明123(Rh123)双染与流式细胞术(FCM),研究了不同盐度胁迫(10、15、20、25、30、35)对精子质量的影响,并对受胁迫精子进行授精实验。结果显示,随着盐度升高,精子的存活率、运动时间和活力先升高后降低;葡萄牙牡蛎精子的适宜盐度为20～35,活力较高的盐度范围为25～35;通过双染和FCM检测了精子质膜完整性和线粒体活性,发现盐度胁迫先对精子质膜造成损伤,后对线粒体活性造成伤害;低盐胁迫(10、15)对精子损伤严重,胁迫15 min时质膜受损比例高达67.26%±2.35%。人工授精结果显示,低盐胁迫下精子受精率和卵裂率显著降低,且卵裂阶段出现畸形及分裂停止等现象,表明精子质量不仅严重影响了受精过程,还对卵裂造成一定影响。本实验不仅探究了PI/Rh123双染和FCM检测牡蛎精子质膜完整性和线粒体活性的可行性,还结合显微观察全面评价了精子质量,为牡蛎精子质量的研究提供了理论基础。     

浙江省健跳港蛎江滩牡蛎多样性和资源量的时空变化

调查了浙江省三门县健跳港蛎江滩牡蛎礁中牡蛎的种类、资源量和幼体补充量，并分析了牡蛎的遗传多样性及资源量和幼体补充量的时空变化特征。结果显示，蛎江滩牡蛎礁中分布有6种牡蛎，即近江牡蛎(Crassostrea ariakensis)、熊本牡蛎(C.sikamea)、福建牡蛎(C.angulata)、香港牡蛎(C.hongkongensis)、猫爪牡蛎(Talonostrea talonata)和长牡蛎(C.gigas)。在牡蛎群落组成中，熊本牡蛎、香港牡蛎和近江牡蛎的占比分别为92.92%、5.00%和2.08%,其他3种牡蛎仅在定性调查中发现。夏初和秋季牡蛎平均密度分别为(97±40)个·m^-2和(1 770±467)个·m^-2;随着潮间带高程的降低，熊本牡蛎密度逐渐降低，而近江牡蛎和香港牡蛎的密度逐步升高。牡蛎幼体附着与补充开始于6月初、结束于10月初左右，补充量呈现显著的时空变化特征；熊本牡蛎在整个礁区均有较高的补充量，近江牡蛎和香港牡蛎则在中、低礁区补充量较高。结果表明，蛎江滩牡蛎礁中的牡蛎群落具有较高的物种多样性。未来需进一步研究泥沙淤...     

低渗诱导太平洋牡蛎三倍体以及与其他诱导方法的比较

通过低渗方法抑制受精卵第二极体(PB2)的释放,诱导太平洋牡蛎(Crassostrea gigas)三倍体.在水温为23～25℃的条件下,分别采用不同低渗海水(盐度分别为4、6、8、10、12、14、16),在不同处理时机,即第1个第一极体(PB1)出现、30%的PB1出现、40%的PB1出现、50%的PB1出现以及第1个第二极体(PB2)出现时,对太平洋牡蛎受精卵进行10 min、15 min、20 min、25 min持续处理.处理后,于正常盐度海水中孵化,收集D形幼虫,通过流式细胞仪进行倍性测定,确定最佳诱导条件.结果表明,当太平洋牡蛎受精卵PB1出现40%时,在盐度为8的低渗海水中,持续处理15 min,得到最高三倍体诱导率为(89.16±1.39)%.与对照组相比,低渗诱导获得的三倍体幼虫表现出明显的生长优势.将低渗方法同高温(32℃)、低温(2℃)和6-DMAP(450μmo1/L)处理等多倍体诱导方法进行比较,结果显示,低渗组的卵裂率和孵化率显著高于高温、低温和6-DMAP诱导的太平洋牡蛎三倍体(P＜0.05).6-DMAP诱导的三倍体率虽略高于低渗诱导,但差异不显著(P＞0.05).通过综合评价指数(Ie)比较,认为低渗诱导多倍体的方法比高温、低温和6-DMAP诱导具有明显的优势.     

盐度骤降对近江牡蛎和长牡蛎能量收支的影响

采用实验生态学方法，研究了盐度骤降(盐度分别为10、20，自然海水为对照组)对近江牡蛎(Crassostrea ariakensis)和长牡蛎(Crassostrea gigas)生理代谢的影响。结果显示，盐度骤降对长牡蛎和近江牡蛎的耗氧率(OR)、排氨率(NR)、排粪率(FER)均有显著影响(P<0.05)，且2种牡蛎对盐度骤降的响应存在差异。长牡蛎的耗氧率、排氨率、摄食率(IR)和排粪率均在自然海水中最高，低盐对长牡蛎产生不同程度的胁迫。而近江牡蛎在盐度为20时的上述指标显著高于其他盐度处理组。通过测定不同规格牡蛎的代谢率，建立2种牡蛎在不同盐度下的代谢率与规格(湿重)的回归方程，该方程符合幂指数：Y=aWb。通过归一化处理，比较湿重在20 g时，2种牡蛎的能量分配差异。盐度为20时，近江牡蛎的呼吸代谢较强(呼吸能占16.1%)，而长牡蛎在自然海水中呼吸代谢较强(呼吸能占7.8%)，2种牡蛎的呼吸代谢均在盐度为10时较低，其中，长牡蛎呼吸能仅占0.7%。研究表明，盐度骤降对长牡蛎的影响大于近江牡蛎，小幅度的盐度骤降对近江牡蛎的能量代谢有一定的促进作用。本研究从能量代谢角度，探讨了近江牡蛎和长牡蛎对盐度骤变的响应，可为牡蛎礁修复工作和养殖生产过程中的品种选择提供参考。     

温度和盐度急性胁迫对岩牡蛎存活及免疫指标的影响

为探究岩牡蛎(Crassostreanippona)对不同温度和盐度急性变化的适应性,本研究通过将岩牡蛎从暂养环境(27℃,盐度32)直接转移至5个温度梯度(15℃、19℃、23℃、31℃、35℃)和6个盐度梯度(16、20、24、28、36、40),研究了急性胁迫对岩牡蛎超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、碱性磷酸酶(AKP)、溶菌酶(LSZ)活性、丙二醛(MDA)含量及存活的影响。温度实验结果显示,15～31℃组岩牡蛎存活率均高于71.11%,35℃组存活率最低(34.44%),而15℃组免疫指标活性均受到明显抑制, MDA含量在144 h时达到最大值(46.93 nmol/mg prot);温度19℃、23℃、31℃组SOD、CAT、AKP和LSZ活性及MDA含量受应激显著上升后均逐渐恢复至对照水平。盐度实验结果表明, 40组盐度变化对岩牡蛎存活抑制明显,显著低于其他处理组(P0.05),适宜岩牡蛎存活的盐度变化为20～36组;盐度16组SOD、CAT、AKP和LSZ活性144 h时均显著低于对照组(P0.05), MDA含量与胁迫时间成正相关最终达到最大值(45.76 nmol/mg prot)。研究表明,温度和盐度突变均会对岩牡蛎5项免疫指标和存活率产生显著影响,岩牡蛎对温度19～31℃组和盐度20～36组突变具有较强的适应能力。     

瓦氏黄颡鱼精子的生理特性及其超低温冷冻保存的初步研究

对瓦氏黄颡鱼(Pelteobagrus vachelli)精子在不同盐度和pH下的精子活力进行观察,同时研究了精子在4种不同稀释液与2种不同浓度抗冻剂组成的保存液中的超低温冷冻保存,并开展了冻精的授精实验。结果表明,瓦氏黄颡鱼精子浓度为(2.035±0.179)×1012cell·mL-1,在盐度为5.8、pH为7.17时,精子的活力都高达95%。以A液作为稀释液、10%甲醇作为抗冻剂时,冷冻保存精子效果最好,解冻后精子活力为(81.7±0.9)%。用解冻后的精子进行人工授精,获得的受精率为(88.4±2.1)%,孵化率为(74.0±0.8)%;而鲜精受精率为(91.0±0.8)%,孵化率(82±1.6)%,冻精与鲜精均无显著性差异。人工授精实验证明了解冻后的精子能正常用于该鱼的人工繁殖。     

熊本牡蛎多嵴和无嵴品系F1生长性状的群体选育

为了改良熊本牡蛎(Crassostrea sikamea)生长性能,以多嵴品系和无嵴品系熊本牡蛎为材料,以壳高为指标,按照10%留种率,采用截头法进行了两个品系的群体选育研究,结果表明:选择反应及现实遗传力随着个体增大而增加,且多嵴品系选择反应略高于无嵴品系。对于多嵴品系而言,幼虫期、稚贝期、养成的选择反应及现实遗传分别为0.33±0.04、0.19±0.02,0.46±0.03、0.26±0.02,0.63±0.11、0.36±0.07;无嵴品系的分别为0.30±0.04、0.17±0.02,0.43±0.03、0.25±0.02,0.58±0.10、0.33±0.06。经过1周年的养成,多嵴品系和无嵴品系的壳高遗传改进量分别为8.41%、7.71%。由此可见,两个品系的现实遗传力相对较高,说明存在着一定程度的遗传变异。研究为熊本牡蛎遗传改良及新品系培育提供了理论与实践基础。     

长牡蛎3种壳色家系间杂交子代生长和存活比较

为了利用杂种优势培育长牡蛎优良新品种,实验以3种不同壳色长牡蛎家系(白色/W、黑色/B、紫色/P)为材料,采用3×3完全双列杂交法,建立了3个自交组合和6个正反杂交组合,分析了各实验组幼虫期和养成期的生长、存活以及杂交子代的杂种优势。结果表明,浮游幼虫期,杂交组PB表现出显著的生长优势;与自交组相比,各杂交组均有着较高的幼虫存活率;在幼虫存活率方面,所有杂交组均有较高的杂种优势率。在养成期,紫壳色自交组的壳高显著大于白壳色和黑壳色自交组;6个杂交组中,PB的壳高生长最快,BP次之,PW、WP的生长最慢;各杂交组与自交组的成活率差异均不显著;杂交组BP及其反交组PB的壳高、壳长、总重和存活率的杂种优势率分别介于3.71%~15.27%、-2.00%~13.10%、11.23%~41.56%、-2.77%~9.83%,其他4个杂交组在整个养成阶段没有表现出杂种优势。     

Summer mortality of hatchery-produced Pacific oyster spat (Crassostrea gigas). I. Estimation of genetic parameters for survival and growth

The multidisciplinary project “MOREST” aims to improve our understanding of causes of summer mortality in Crassostrea gigas juveniles in France and to reduce its impact on oyster production. As part of the MOREST project, 43 full-sib families nested within 17 half-sib families were produced, planted out and tested in 3 sites during summer 2001 to assess to what extent genetic variability exists for this trait. A strong genetic basis was found for survival as narrow-sense heritability estimates ranged from 0.47 to 1.08, with higher values in sites where summer mortality was higher. Genetic correlations across sites were positive and very high for survival, indicating no genotype by environment interaction. In contrast, lower genetic variation was observed for growth in all sites. Finally, genetic correlations between growth and survival were low, in all sites. Selective breeding in a single site should therefore be an efficient means of improving survival in oysters less than one year old along the French Atlantic and Channel coastlines with only very limited effects on growth. As yield mostly depends on survival and growth, this approach should significantly improve harvestable yield. Possible reasons why a high genetic variance for survival appears to be maintained in wild populations are discussed.     

Reproductive cycle and biochemical composition of the Zhe oyster Crassostrea plicatula Gmelin in an eastern coastal bay of China

The reproductive cycle and biochemical composition of the oyster Crassostrea plicatula in Rushan Bay, an eastern coastal bay of China, were investigated between March 2004 and February 2005 in relation to environmental factors (temperature, salinity and chlorophyll a). Histological analysis, combined with oocyte examination and measurements of protein, glycogen and lipid levels and RNA/DNA ratio from gonad, adductor muscle and mantle tissue of each sex were performed. The gametogenic cycle of C. plicatula comprised two phases: a resting phase (November-February) and reproductively active phase, including ripeness and spawning, during the rest of year. Gametogenesis in spring and summer was associated with an increase in protein and lipid contents in the ovaries, and took place at the expense of reserves (mainly glycogen) accumulated previously during winter. The RNA/DNA ratio is a good indicator of sexual maturity in the ovary; the increasing RNA/DNA ratio in the ovary appears to show the rising synthetic activity of vitellin within the ovary. Spawning occurred at the moment when water temperature and chlorophyll a levels were highest, and salinity lowest throughout the year. High variation in glycogen contents during storage and gametogenic development suggests that carbohydrates are the main respiratory substrate in this species. The results demonstrated that C. plicatula may be considered a conservative species in gametogenic pattern.     

Direct and indirect responses to selection on individual body weight in the Pacific oyster (Crassostrea gigas)

Three experiments were performed to examine the heritability of body weight among adult Pacific oysters (Crassostrea gigas) evaluated in Yaquina Bay, Oregon, USA, and to determine if selection on individual body weight could result in changes in offspring survival and yield. The first two experiments utilized midparent-offspring regressions to estimate the heritability (h²) of adult oyster body weight and the coheritability (h_x,y) between adult midparent body weight and offspring performance, including juvenile average body weight, survival and yield as well as adult survival and yield. In Experiment 1 both parents and offspring were evaluated in an “upriver” environment in Yaquina Bay. In Experiment 2 parents were evaluated in a “downriver” environment, while offspring were evaluated in an “upriver” environment. Experiment 3 contrasted average body weight, survival, and yield of offspring (evaluated upriver) derived from three large sires and three small sires mated to the same five females that were chosen at random (all parents evaluated downriver). In Spring 2002, 12 full-sib families from Experiment 1, 19 families from Experiment 2, and 26 families from Experiment 3 were stocked into lantern nets and suspended in Yaquina Bay. Measurements of yield (kg tier^− 1), average body weight (g), and survival (%) were recorded after one and two growing seasons in the field. Heritability estimates for adult body weight at harvest ranged from 0.313 (± 0.083) in Experiment 1 to 0.003 (± 0.073) in Experiment 2. In Experiment 3, average body weight did not differ between offspring derived from large sires and offspring derived from small sires (P = 0.47). Significant negative coheritability estimates were observed between adult midparent body weight and offspring survival in both Experiment 1 and Experiment 2. Significant negative coheritability estimates between adult midparent body weight and offspring yield were observed in Experiment 2 but not in Experiment 1. In Experiment 3, offspring derived from large sires had significantly lower survival and yield than offspring derived from small sires. These results show adult oyster body weight to be heritable but also subject to site-specific adaptation such that selection in the downriver Yaquina Bay environment was ineffective at changing average body weight in the upriver environment. Negative coheritability estimates between performance traits indicate that adult oyster body weight may be a poor indirect measure of oyster yield potential, and that selection solely for increased body weight could lead to a decrease in offspring yield.     

Setting of a procedure for experimental incubation of Pacific oyster (Crassostrea gigas) embryos

Standardized experimental protocols designed to study Pacific oyster (Crassostrea gigas) gamete quality have not been previously published. Gamete sampling variations and confounding effect of interacting factors result in large variations between replicates, decreasing the effects of the studied factors. This work aims at defining a standardized procedure for incubation of oyster embryos designed for experimental purposes.In a first phase, four experiments were developed to improve embryo sampling and handling. They showed that a minimum of 50 embryos must be counted to decrease the variation between counting replicates. For reliable results, sampling must be carried out from 0 to 7 min after a careful agitation of seawater containing embryos (salinity: 35.6‰). Compared to control and a 30 cm one, the D-larval yield of embryos submitted to a 1 m fall was significantly reduced, showing the limited effect of mechanical disturbance on Pacific oyster oocytes survival. Then, maintaining 2.2 million embryos in 250 ml seawater for a two hours period resulted in a decrease of D-larval yield which could not be explained by a decrease of O₂ content and pH of seawater.The second phase of this work included a set of five experiments, defining experimental incubation conditions. A higher larval yield was observed using 1.8 l beakers and 150 l tanks compared to 0.3 ml microtiter plates and 1 l fish egg incubators. Higher larval yields were recorded when embryo density ranged from 5 to 100 ml^− 1, compared to values between 500 and 2000 ml^− 1. Compared to controls (no antibiotic or no presence of light), no changes of D-larval yield were observed by adding 20 ppm chloramphenicol or by maintaining embryos in total darkness. Then, a significant decrease of pH and O₂ content was observed during the incubation period. However, these changes could not be considered as limiting for Pacific oyster embryo development.In conclusion, experimental incubation conditions have been defined in this study: 30 embryos ml^− 1 incubated in 1.8 l beakers without antibiotic and regardless of light intensity, when not higher than 500 lux. The mean coefficients of variation observed between tank replicates ranged from 13.1 to 16.5%. The standardized incubation procedure described in this work will help to study quality variations of Pacific oyster gametes.     

Effects of mass and position of artificial fouling added to the upper valve of the mangrove oyster Crassostrea rhizophorae on its growth and survival

We ran an experiment on mangrove oysters Crassostrea rhizophorae to evaluate the effects of adding different masses of artificial fouling to the upper valve, either to the umbo region or the ventral edge of the shell. Growth and survival were quantified after a 30 d period in suspended culture in the La Restinga Lagoon, Venezuela. The artificial fouling was cement weighing 0.5, 1, 2 or 3 fold the mass of the upper valve. No fouling was added to a control group. Fouling mass, but not the position of the artificial fouling, affected growth in shell length. However, only the heaviest fouling (3 times the mass of the upper valve) had a significant effect. In contrast, there was no affect of either fouling mass or position on tissue growth. Finally, our data indicated that mortality could be affected by the position where we added artificial fouling (greater mortality when fouling was added to the ventral edge of the shell), but not by fouling mass. Our study indicates it is unlikely that the levels of natural fouling that develop on oysters in suspended culture would be sufficient to affect either growth or survival.     

Susceptibility by amberjack (Seriola dumerili), yellowtail (S. quinqueradiata) and Japanese flounder (Paralichthys olivaceus) to Neobenedenia girellae (Monogenea) infection and their acquired protection

This study investigated: 1) susceptibility differences to infection by Neobenedenia girellae (Capsalidae) between amberjack Seriola dumerili (Carangidae), yellowtail S. quinqueradiata and Japanese flounder Paralichthys olivaceus (Paralichthyidae); 2) growth and egg production of N. girellae on each fish species; 3) acquired protection of each fish species against this parasite. The number of N. girellae on S. dumerili was significantly higher than on S. quinqueradiata and P. olivaceus when these fishes were exposed to oncomiracidia in the same aquarium. Neobenedenia girellae growth on S. dumerili was fastest and, thus the number of eggs laid by parasites on S. dumerili was greater than on the other two species. Seriola dumerili and P. olivaceus, which were previously infected with N. girellae and treated by freshwater bath, acquired partial protection against re-infection by N. girellae. The relative re-infection of three S. dumerili individuals out of eleven individuals was markedly low compared with the initial infection, and the relative initial infection and re-infection on two P. olivaceus out of eleven individuals was markedly low. The results of this study could be useful to control N. girellae infections when cultivating S. dumerili, S. quinqueradiata and P. olivaceus.     

An insulin-like system involved in the control of Pacific oyster Crassostrea gigas reproduction: hrIGF-1 effect on germinal cell proliferation and maturation associated with expression of an homologous insulin receptor-related receptor

The putative involvement of insulin-like peptides in the control of the reproduction of the Pacific oyster Crassostrea gigas was investigated using different approaches. In conjunction with a monthly histological analysis of the oyster reproductive cycle, in vitro biological effects of the human recombinant IGF-1 (hrIGF-1) on dissociated germinal cells were mesured over 1 year using [³H]-thymidine and [¹⁴C]-amino acid mixture as tracers for DNA and protein synthesis. DNA synthesis was stimulated by hrIGF-1 in November (114 ± 11% for 10^− 7M), December (46 ± 6% for 10^− 7 M) and January, which was identified as the highest gonial mitosis period. A clear dose-effect was observed in January with a maximum activation of 68 ± 7% for 10^− 12 M. Germinal cell protein synthesis was also stimulated in March (20 ± 1% for 10^− 10 M), April (41 ± 5% for 10^− 13 M), May (25 ± 4% for 10^− 13 M), and by almost all of hrIGF-1 doses in June (21.5 ± 2% for 10^− 13 M) and July (34 ± 1% for 10^− 13 M). This suggests the involvement of insulin-like substances in gonadal tubule rebuilding (December), as well as in the development of germinal cells (March, April), and in the summer maturation of gametes (May, June, July). These insulin-like effects conform with the expression pattern of the recently identified C. gigas insulin receptor-related receptor (CIR): It appeared highly expressed in the gonadal area during gonial mitosis phase, but also in maturating oocytes, suggesting the involvement of an insulin-like system in gonial proliferation and maturation. Moreover, CIR showed differential expression during embryogenesis and larval developmental stages. The expression of maternal CIR during the embryonic and early larval development, followed by the increasing zygotic CIR expression from D larvae to 11-day-old veliger larvae, then a decrease until metamorphosis, also suggest that insulin-like peptide is involved in organogenesis.     

太平洋鳕RAG1和IgM基因荧光定量PCR方法的建立及初步应用

为研究太平洋鳕发育早期特异免疫系统形成的机制,通过RAG1和IgM基因的转录水平衡量特异免疫系统的发育特点.根据GenBank中RAG1和IgM的序列信息,分别设计1对特异引物,从太平洋鳕头肾中扩增得到RAG1和IgM的基因片段.将所获基因片段分别插入到克隆载体pMD18-T中,从而构建太平洋鳕RAG1和IgM基因的质粒标准品.建立并优化太平洋鳕RAG1和IgM基因绝对荧光定量PCR方法.为进一步验证该方法的可靠性,分别利用绝对定量和相对定量检验目的基因在太平洋鳕早期发育过程不同组织内的表达差异.以优化后的绝对荧光定量PCR方法检测不同发育时期太平洋鳕RAG1和IgM的表达情况.结果显示,RAG1的回归方程为y=-3.266x+33.77,回归系数R2=0.996;IgM的回归方程为y=-3.119x +27.61,回归系数R2 =0.998.绝对定量和相对定量结果在基因转录趋势上显现出一致性,即RAG1基因在胸腺和头肾中表达,且在胸腺中的表达量显著高于头肾中的表达量,在肝脏和脾脏中无表达;IgM基因在胸腺、头肾、肝脏和脾脏中均有表达,其中脾脏中表达量最高,其次是头肾.RAG1基因在太平洋鳕发育早期的表达水平很低,到61日龄(days posthatching,dph)至95 dph表达量显著提高;IgM基因在早期表达水平同样很低,到33 dph至61 dph才有明显表达,在95 dph时表达量显著提高.研究表明,本实验方法可靠,特异性较强,可成功对目标基因转录水平进行检测.     

Evaluation of three seaweeds Gracilaria bursa-pastoris, Ulva rigida and Gracilaria cornea as dietary ingredients in European sea bass (Dicentrarchus labrax) juveniles

The aim of this study was to evaluate the inclusion of three seaweeds Gracilaria bursa-pastoris (GP), Ulva rigida (UR) and Gracilaria cornea (GC) as dietary ingredients on the performance, nutrient utilisation and body composition of European sea bass juveniles. Six experimental diets were formulated to replace 5% (GP-5, UR-5, and GC-5 Diets) and 10% (GP-10, UR-10 and GC-10 Diets) fish protein hydrolysate (CPSP) by each of the three seaweeds. A control diet was used, without inclusion of any seaweed. Diets were fed to duplicate groups of 25 European sea bass (Dicentrarchus labrax) juveniles (IBW = 4.7 g) for 10 weeks. Growth performance was only significantly reduced (P < 0.05) in fish fed the GC-10 diet, whereas the feed conversion ratio increased significantly in those fish. The apparent digestibility coefficients of dry matter and lipid were significantly lower in fish fed diet GC-10 relative to those fed the control diet. Carcass composition was similar among treatments, although fish fed GC-10 exhibited significantly higher ash content.The results obtained in this study suggest that the inclusion of G. bursa-pastoris (GP) and U. rigida (UR), up to 10%, can be considered as very interesting ingredients in diets for sea bass juveniles, as no negative consequences on growth performance, nutrient utilization or body composition were observed. On the other hand, the inclusion of G. cornea (GC) should be limited to 5% of the diet.