香港巨牡蛎和长牡蛎幼虫及稚贝的表型性状

为了评估香港巨牡蛎和长牡蛎在北方沿海的早期表型性状,于2010年7月,以2009年6月在青岛繁育的两种牡蛎为材料,在大连研究了温度(Mt:(22±1.0)℃及Ht:(28±1.0)℃)、盐度(S₂₀:20±1.0及S₃₀:30±1.0)及中间育成环境(ID:室内及OD:室外)对两种牡蛎幼虫及稚贝表型性状的影响。结果表明:香港巨牡蛎壳宽显著大于长牡蛎(P<0.05),壳高及怀卵量显著小于长牡蛎(P<0.05),壳长、鲜重及壳重两者间无显著差异(P>0.05)。在温度和盐度相同情况下,长牡蛎卵径、受精率、孵化率及D形幼虫均大于香港巨牡蛎;香港巨牡蛎幼虫浮游前期生长较慢,而后快于长牡蛎。两种牡蛎幼虫存活能力在15日龄时高温组>中温组;相同温度下,香港巨牡蛎中盐组>高盐组,长牡蛎高盐组>中盐组。幼虫变态期间,较低的温度延迟了变态时间,降低了变态率,使得两种幼虫变态规格大型化。温度是影响幼虫生长、存活、变态的最主要因素,其次为盐度,交互作用几乎尚未起到作用。中间育成阶段,室外比室内培育效果更好,且香港巨牡蛎稚贝壳高在60日龄以后显著大于长牡蛎(P<0.05),环境是影响稚贝生长的最主要因素;无论室内还是室外两种牡蛎稚贝的存活率均在90%以上,且各实验组间无显著差异(P>0.05)。     

盐度对长牡蛎和近江牡蛎及其杂交稚贝生长和存活的影响

2011年8月以长牡蛎自繁组GG(Crassostrea gigas♀×C.gigas♂),近江牡蛎自繁组AA(C.ariakensis♀×C.ariakensis♂)、正交组GA(Crassostrea gigas♀×C.ariakensis♂)、反交组AG(C.ariakensis♀×C.gigas♂)为实验材料,开展了稚贝对盐度的适应性研究。结果发现长牡蛎的最适生存盐度为15—35,最适生长盐度为25—35;近江牡蛎的最适生存盐度为10—25,最适生长盐度为20—25;GA的最适生存盐度为15—30,最适生长盐度为15—30,AG的最适生存盐度为20—30,最适生长盐度为20—25。GG对低盐度敏感,AA对高盐度敏感,AG具有高盐度存活的杂种优势,在盐度30时,中亲杂种优势HG×A为13.32,单亲杂种优势HGA和HAG分别为1.89和27.88,在盐度40时,HAG上升到400,GA和AG都不具有生长优势。杂种稚贝对盐度适应介于双亲之间,且表现出一定程度的父系遗传特点。     

香港牡蛎Foxl2基因克隆分析与表达模式研究

本研究旨在对香港牡蛎Foxl2基因进行克隆、序列分析及建立其在香港牡蛎组织中的表达模式谱。以GenBank数据库中已上传的太平洋牡蛎Foxl2基因序列为模板,设计特异性引物扩增获得香港牡蛎Foxl2基因;应用生物信息学方法,系统分析和预测香港牡蛎Foxl2基因的遗传进化、蛋白质的理化性质和结构特征;并应用QRT-PCR技术对Foxl2在香港牡蛎组织中的表达进行了差异分析。结果表明:香港牡蛎Foxl2基因编码区全长1 104 bp,共编码氨基酸367个。多重序列比较分析显示香港牡蛎Foxl2核苷酸序列相似性与太平洋牡蛎、美洲牡蛎、大珠母贝、栉孔扇贝、虾夷扇贝、皱纹盘鲍相应序列的相似性分别为97%、83%、71%、76%、75%、75%,系统进化树分析显示在不同物种以及进化的过程中Foxl2基因具有较高的保守性。Foxl2蛋白质信息学分析结果表明该蛋白呈碱性,N端无信号肽存在,位于胞核中,含有FH等结构。定量分析结果显示:Foxl2在香港牡蛎3种组织中有表达,其中表达量最高的是性腺,其次是唇瓣,鳃中相对表达量最低。本研究成功地克隆了香港牡蛎Foxl2基因,并研究了其在香港牡蛎8种组织中的表达变化规律,为今后阐明其在香港牡蛎性腺发育进程中的功能提供了理论依据。     

硇洲岛牡蛎相关细菌抗菌筛选及系统发育分析

以8个敏感菌株作为指示菌,采用管碟法对分离自湛江硇洲岛（20°52′N～20°56′N,110°33′E～110°38′E）潮汐带香港巨牡蛎(Crassostrea hongkongensis)中的72株细菌的发酵液进行抗菌筛选,并对阳性菌株进行基因组DNA提取、16S rRNA基因PCR扩增和序列测定,继而进行系统发育分析。抗菌实验结果表明,受试菌株中有23株菌的发酵产物具有抗菌活性（阳性率31.9%）,其中有5个菌株（JSM 111024、JSM 111027、JSM 111029、JSM 111076、JSM 111083）具有较强的抗菌活性。基于16S rRNA基因序列的系统发育分析表明,这23株菌具有较高的类群多样性和物种多样性,属于3个系统发育群/门(Alpha Proteobacteria、Gamma Proteobacteria、Bacteroidetes)中的8个科(Aeromonadaceae、Flavobacteriaceae、Halomonadaceae、Idiomarinaceae、Phyllobacteriaceae、Pseudoalteromonadaceae、Shewanellaceae、Xanthomonadaceae)的8个属(Idiomarina、Halomonas、Myroides、Nitratireductor、Oceanimonas、Pseudoalteromonas、Shewanella、Wohlfahrtiimonas),可分为11个物种。优势类群为Gamma Proteobacteria亚门（14株）,其中优势属为Oceanimonas属（6株）;第二大类群为Bacteroidetes门（7株）,都属于Flavobacteriaceae科的Myroides属。具有较强抗菌活性的5个菌株中,有4个菌株（JSM 111024、JSM 111027、JSM 111029、JSM 111083）属于Alpha Proteobacteria 亚门Phyllobacteriaceae科〖WTBX〗Oceanimonas属,而菌株JSM 111076属于Gamma Proteobacteria 亚门Aeromonadaceae科Nitratireductor属。     

温度对巨膜长蝽生长发育与繁殖的影响

【目的】近年来巨膜长蝽Jakowleffia setulosa (Jakovlev)由稳定种群的荒漠昆虫上升为暴发性发生并迁移至农区危害的农业害虫。由于缺乏巨膜长蝽生物学基础研究,给监测预警及综合防治带来许多困难。本研究旨在探明温度对巨膜长蝽发育和繁殖的影响。【方法】本实验以白茎盐生草Halogeton arachnoideus Moq.为寄主植物,分别在7个恒温（13,18,23,28,33,35和37℃）下研究了温度对巨膜长蝽各虫态平均发育历期、发育速率、成活率、产卵量和种群参数的影响。【结果】在13~37℃范围内,各虫态的发育历期随温度的升高而缩短,13℃完成一个世代发育需要82.63 d,37℃只需要14.61 d。各虫态的发育速率（V）与温度（T）的关系均符合多项式回归模型,且极显著相关。采用直接最优法分析,巨膜长蝽世代的发育起点温度为8.30℃,有效积温为555.77日·度,在宁夏一年可发生2代。巨膜长蝽世代存活率均在33℃时最高,为35.44%。28～37℃之间单雌产卵量较多,达到10粒以上,通过方程拟合得到理论上雌虫产卵最适温度为34.59℃,产卵量最高可达到16.73粒/雌;种群趋势指数在13℃为零,18～23℃小于1,种群呈负增长;28～37℃大于1,且33℃时最高,为2.77;净增殖率、内禀增长率及周限增长率均在37℃时最高,分别为7.24, 0.3912和1.4787;种群世代周期以13℃时最长,为45.8936 d,37℃时最短,为5.0608。【结论】巨膜长蝽在28～35℃下存活率、增长指数和繁殖力较高,说明该温度范围是巨膜长蝽生长发育和繁殖的适宜温度。本研究为有效开展巨膜长蝽的监测预报及综合防控提供了科学依据。     

长牡蛎精子超低温冷冻后超微结构损伤研究

采用程序降温仪分步降温冷冻保存长牡蛎（Crassostrea gigas）精液,并用扫描电镜、透射电镜研究了精子的超微结构损伤。超低温冷冻保存后长牡蛎精子的运动率、受精率及孵化率与鲜精无显著差异。鲜精中84.5%的精子形态结构正常,冻精中73%的精子形态结构正常。形态结构正常的精子表现为顶体、质膜、线粒体与鞭毛结构完整、染色质形状规则,顶体、线粒体及中心粒结构正常,鞭毛形态完整、微管结构清晰;形态结构异常的精子表现为顶体脱落、解体,精子头部质膜膨胀、破裂、染色质肿胀、破裂、解体,线粒体移位、脱落、膨胀,嵴退化或消失,鞭毛弯折、断裂,微管解聚。结果显示,以10% DMSO为抗冻保护剂,HBSS溶液为稀释液,1:4的稀释比例,添加海藻糖,采用分步降温法冷冻保存,对长牡蛎精子具有较好的抗冻保护作用,合适的冻存方法可以有效的保护太平洋牡蛎精子冷冻过程中结构损伤。研究有助于长牡蛎种质资源的收集保存及应用。     

香港长洲岛野生植物物种多样性与植被的研究

长洲岛共有野生维管植物237种,隶属于73科184属.植被类型以次生常绿阔叶林为主,其次为灌丛群落.植物区系以热带亚热带植物地理成分占优势,热带性质明显.该岛的植物区系与邻近岛屿植物区系具有极大的相似性,在香港植物区系组成中占有一定地位.由于人为活动和外来植物的影响,其植物多样性及植被的保护应引起注意.     

温度、盐度及交互作用对僧帽牡蛎(Saccostrea cucullata)呼吸排泄的影响

采用实验生态学方法研究了温度(13、18、23、28、31、34℃)和盐度(18、23、28、33)对僧帽牡蛎(Saccostrea cueullata)耗氧率和排氨率的影响,并分析了二者交互作用对呼吸排泄的影响。结果表明,不同温度条件下,僧帽牡蛎的耗氧率变化范围为0.43-3.15mg g~(-1)h~(-1),排氨率的变化范围为72.62-1245.77μg g~(-1)h~(-1),方差分析显示,温度和盐度对僧帽牡蛎耗氧率(F=60.440,P0.01)和排氨率(F=61.083,P0.01)的影响均极显著;二者的交互作用对僧帽牡蛎耗氧率(F=4.907,P0.01)和排氨率(F=11.412,P0.01)影响也极显著,耗氧率(Y)、排氨率(Y)与温度(T)和盐度(S)之间的二元线性回归方程为y=a+bT+cS+dT×S。在一定温度范围内(13-31℃)僧帽牡蛎单位体重耗氧率(Y)与温度(T)呈正相关符合指数方程Y=ae~(bT),其中a的取值范围为0.1378-0.2117,b的取值范围为0.0757-0.0912;单位体重排氨率也与温度呈正相关,符合上述指数方程,其中a的取值范围为20.364-30.634,b的取值范围为0.1004-0.1341。不同温度和盐度条件下,僧帽牡蛎呼吸排泄的氧氮比在2.20-7.78之间,平均值为3.81。     

巨膜长蝽的形态特征和生物学特性

【目的】近年来巨膜长蝽Jakowleffia setulosa(Jakovlev)由以往稳定种群的荒漠昆虫上升为暴发性发生并迁移至农区危害的农业害虫。由于缺乏对巨膜长蝽基础生物学的相关研究,对其开展监测预警和农业防治存在困难。开展巨膜长蝽个体生物学研究,填补研究空白对于有效监测预报和防控提供可靠的依据,同时能为下一步分析巨膜长蝽成灾规律提供基础数据。【方法】本文通过室内观察和野外调查相结合,系统阐明该种形态特征、生活史、习性和行为等生物学特性。【结果】巨膜长蝽在宁夏中部干旱带一年发生2代,5月中旬和10月中下旬是两个发生高峰期,6月中旬至8月中旬以成虫进入滞育状态,11月下旬以成虫越冬;雌成虫的平均寿命为(32.14±2.34)d,雄成虫的平均寿命为(28.00±3.13)d,雌雄性比为1︰1.9,有多次交尾多次产卵的习性,每头雌虫抱卵量10粒左右,平均产卵量10~15粒左右;食性杂,群居危害,喜食白茎盐生草(Halogeton arachnoideus)等沙生草本植物的种子。【结论】该虫具有以成虫形态越冬和滞育、迁移性强等生物学特性。在生态环境发生较大变动后,能够经历逆境后迅速恢复种群并迁移危害。宁夏中南部地区种植结构调整,极大改变了该地区的荒漠生态环境,促使巨膜长蝽在经过夏季高温滞育恢复种群后无法获得足够野生寄主的情况下,大规模迁入农田系统危害。     

硇洲岛潮汐带牡蛎相关可培养细菌多样性

[目的]了解南海硇洲岛潮汐带香港巨牡蛎(Crassostrea hongkongensis)相关可培养细菌的多样性.[方法]应用纯培养法分离样品中的细菌(含放线菌),采用基于16S rRNA基因序列的系统发育分析方法研究这些分离菌株的生物多样性.[结果]用补充0-25％(W/V) NaC1的MA、MH和NA培养基从样品中分离到102株细菌,在形态观察和部分生理生化实验结果的基础上去冗余,选取74个代表性菌株进行基于16S rRNA基因序列的系统发育分析.结果表明,这些菌株归为38个物种,属于4个大的系统发育类群(Gamma-Proteobacteria,Alpha-Proteobacteria,Bacteroidetes,Firmicutes)的16个科、18个属.大多数菌株属于Gamma-Proteobacteria亚门(45株,60.8％),其余依次是Firmicutes门(12株,16.2％)、Bacteroidetes门(11株,14.9％)和Alpha-Proteobacteria亚门(6株,8.1％).大多数菌株与其系统发育关系最密切的有效发表种典型菌株之间(16S rRNA基因序列相似性为94.3％-99.8％)存在一定的遗传差异,其中JSM 111069可能代表Carnobacteriaceae科的潜在新属;菌株JSM 111039和JSM 111085可能分别代表Bacillus属的两个新物种,菌株JSM 111020、JSM 111072和JSM 111090可能分别代表Pseudoalteromonas、Proteus和Idiomarina属的新物种.[结论]南海硇洲岛潮汐带牡蛎中存在较为丰富的原核生物多样性,并潜藏着一定数量的微生物新类群(物种).     

Individual relationship between aneuploidy of gill cells and growth rate in the cupped oysters Crassostrea angulata, C. gigas and their reciprocal hybrids

The Portuguese oyster, Crassostrea angulata, is taxonomically close to the Pacific oyster, C. gigas, but there are clear genetic and phenotypic differences between these taxa. Among those differences, the faster growth of C. gigas compared with C. angulata has often been observed in the field. Crosses between C. angulata and C. gigas were performed to investigate the relationship between growth variation and somatic aneuploidy at the individual level in the two taxa and their reciprocal hybrids. The different progenies were reared in Ria Formosa (Portugal) under standard farming conditions. Growth rate and survival were significantly higher in C. gigas than in C. angulata, and the hybrids showed intermediate performances. Significant differences were also observed in the proportion of aneuploid cells (PAC) and of missing chromosomes (PMC) between the two taxa, C. angulata showing the highest values. Intermediate values of PAC and PMC were observed in the hybrids, supporting additive genetic bases of these parameters. Our results also confirm the negative correlation between somatic aneuploidy and growth rate at the individual level, as previously reported in C. gigas.     

Microsatellite analysis of gynogenetic families in the Pacific oyster, Crassostrea gigas

Five families of gynogenetic diploid Pacific oyster (Crassostrea gigas) were induced by inhibiting the second polar body in meiotic cell division of eggs fertilized with UV-irradiated sperm. Segregation patterns of eight microsatellite loci were investigated in the gynogenetic diploid offspring; the proportion of heterozygous progeny was used to estimate microsatellite-centromere (M-C) distances. Mendelian inheritance was confirmed for the eight loci by examining the genotypic segregation in the control crosses. Three of the eight microsatellite loci showed the existence of null alleles in four control crosses. All gynogenetic offspring only possessed the alleles of the mother, indicating 100% success level for the five families. The M-C recombination frequency estimates ranged from 0.62 to 0.77 (0.72 mean), comparable to those in the oyster based on allozyme markers and suggesting that meiotic gynogenesis does not appear to be a very efficient inbreeding method in the oyster. Recombination frequencies observed were often higher than the theoretical maximum of 0.67, indicating the existence of positive interference after a single chiasma formation in some chromosomes. Information on the positions of centromeres in relation to the microsatellite loci will represent a contribution toward assembly of genetic maps in C. gigas.     

Molecular and histological identification of Marteilioides infection in Suminoe Oyster Crassostrea ariakensis, Manila Clam Ruditapes philippinarum and Pacific Oyster Crassostrea gigas on the south coast of Korea

The oyster ovarian parasite Marteilioides chungmuensis has been reported from Korea and Japan, damaging the oyster industries. Recently, Marteilioides-like organisms have been identified in other commercially important marine bivalves. In this study, we surveyed Marteilioides infection in the Manila clam Ruditapes philippinarum, Suminoe oyster Crassostrea ariakensis, and Pacific oyster Crassostrea gigas, using histology and Marteilioides-specific small subunit (SSU) rDNA PCR. The SSU rDNA sequence of M. chungmuensis (1716 bp) isolated from C. gigas in Tongyoung bay was 99.9% similar to that of M. chungmuensis reported in Japan. Inclusions of multi-nucleated bodies in the oocytes, typical of Marteilioides infection, were identified for the first time in Suminoe oysters. The SSU rDNA sequence of a Marteilioides-like organism isolated from Suminoe oysters was 99.9% similar to that of M. chungmuensis. Marteilioides sp. was also observed from 7 Manila clams of 1840 individuals examined, and the DNA sequences of which were 98.2% similar to the known sequence of M. chungmuensis. Unlike Marteilioides infection of Pacific oysters, no remarkable pathological symptoms, such as large multiple lumps on the mantle, were observed in infected Suminoe oysters or Manila clams. Distribution of the infected Manila clams, Suminoe oysters and Pacific oysters was limited to small bays on the south coast, suggesting that the southern coast is the enzootic area of Marteilioides infection.     

Interspecific hybridization in oysters: Restriction Enzyme Digestion Chromosome Banding confirms Crassostrea angulata × Crassostrea gigas F1 hybrids

The taxonomic status of the two commercially important cupped oysters, Crassostrea angulata, the Portuguese oyster (Lamarck, 1819) and Crassostrea gigas, the Japanese oyster (Thunberg, 1793) has long been in question. The recent observation of the hybridization between C. gigas and C. angulata and the production of fertile F1s led us to search for cytogenetic evidence of both parental genomes in the interspecific hybrids. The cytogenetic characterization of the hybrids was performed by the use of restriction endonuclease treatments. This technique has recently shown the potential for individual chromosome identification by banding in oysters. Chromosomes of C. gigas, C. angulata and their hybrids were treated with two different restriction enzymes (ApaI and HaeIII), stained with Giemsa, and examined for banding patterns. These chromosome markers allowed the parental haploid sets to be identified in the hybrids. The analysis of the banded karyotypes of the interspecific hybrids showed that for each chromosome pair, one of the homologues presented a banding pattern consistent with that of C. gigas and the other homologue presented a banding pattern consistent with that of C. angulata. These cytogenetic results substantiate the reported interspecific hybridization between C. gigas and C. angulata. In view of these results and taking into account the present expansion of C. gigas aquaculture in southern Europe, the question of the need for preservation of pure C. angulata stocks should be raised as only a few populations remain in the south of Spain and Portugal. Recently, changes in the genetic composition of populations in southern Portugal have indeed been observed, showing that human activities have created contact zones between the two taxa while no natural sympatric zones exist in Europe.     

Trophic interactions between two introduced suspension-feeders, Crepidula fornicata and Crassostrea gigas, are influenced by seasonal effects and qualitative selection capacity

The effects of season and qualitative selection capacity on trophic relationships between two sympatric invasive suspension-feeders, Crepidula fornicata and Crassostrea gigas, were investigated in Bourgneuf Bay (France) from January 2003 to June 2004. Carbon and nitrogen stable isotopic deviations, δ¹³C and δ¹⁵N, of common Atlantic slippersnails and Pacific oysters were analysed relative to isotopic composition and availability of end-members.Slippersnail deviations were less variable over the sampling period compared with those of oysters. Significant differences between δ¹³C and δ¹⁵N of C. fornicata and C. gigas were found from winter to early summer, and linked to major isotopic changes in oysters. We identified three distinct seasonal periods: January to March when oysters were ¹⁵N-enriched compared to slippersnails and to themselves at other times of the year, April to June-July when oysters showed a ¹⁵N-depletion and a more marked ¹³C-depletion compared to slippersnails and to themselves at other times of the year, and July-August to December when both species presented similar carbon and nitrogen deviations. Species-specific differences in qualitative selection capability may explain these seasonal differences in isotopic deviations. Whereas the isotopic composition of the indiscriminate suspension-feeding slippersnails reflects the composition of the seston throughout the year, the oyster is capable of qualitative selection. The oyster isotopic compositions are consistent with a facultative activation of selection mechanisms under conditions of qualitative and quantitative food limitation, notably the preferential ingestion and assimilation of the dominant organic source in the suspended pool.We conclude that C. fornicata and C. gigas are trophic competitors only in winter and spring at this site, where detrital end-members are major POM components. These results underscore (1) the importance of long-term (annual) studies in the evaluation of potential trophic competition, and (2) the necessity to include the qualitative selection capacities of suspension-feeders in future interpretations of trophic relationships in marine coastal ecosystems.     

Comparative cryopreservation study of trochophore larvae from two species of bivalves: Pacific oyster (Crassostrea gigas) and Blue mussel (Mytilus galloprovincialis)

Oysters and mussels are among the most farmed species in aquaculture industry around the world. The aim of this study was to test the toxicity of cryoprotective agents to trochophore larvae from two different species of bivalves and develop an improved cryopreservation protocol to ensure greater efficiency in the development of cryopreserved trochophores (14 h old oyster larvae and 20 h old mussel larvae) to normal D-larvae for future developments of hatchery spat production. The cryopreservation protocol producing the best results for oyster trochophores (60.0 ± 6.7% normal D-larvae) was obtained by holding at 0 °C for 5 min then cooling at 1 °C min⁻¹ to −10 °C and holding for 5 min before cooling at 0.5 °C to −35 °C, holding 5 min and then plunging into liquid nitrogen (LN), using 10% ethylene glycol. For mussel experiments, no significant differences were found when cooling at 0.5 °C min⁻¹ or at 1 °C min⁻¹ for CPA combinations with 10% ethylene glycol and at 0.5 °C min⁻¹. Using these combinations, around half of trochophores were able to develop to normal D-larvae post-thawing (48.9 ± 7.6% normal D-larvae).     

Responses of diploid and triploid Pacific oysters Crassostrea gigas to Vibrio infection in relation to their reproductive status

Several Vibrio species are known to be pathogenic to the Pacific oyster Crassostrea gigas. Survival varies according to pathogen exposure and high mortality events usually occur in summer during gametogenesis. In order to study the effects of gametogenetic status and ploidy (a factor known to affect reproduction allocation in oysters) on vibriosis survival, we conducted two successive experiments. Our results demonstrate that a common bath challenge with pathogenic Vibriosplendidus and Vibrio aestuarianus on a mixture of mature, spawning and non-mature oysters can lead to significant mortality. Previous bath challenges, which were done using only non-mature oysters, had not produced mortality. Immunohistochemical analyses showed the affinity of Vibrio for gonadic tissues, highlighting the importance of sexual maturity for vibriosis infection processes in oysters. Mortality rate results showed poor repeatability between tanks, however, in this bath challenge. We then tested a standardized and repeatable injection protocol using two different doses of the same combination of two Vibrio species on related diploid and triploid oysters at four different times over a year. Statistical analyses of mortality kinetics over a 6-day period after injection revealed that active gametogenesis periods correspond to higher susceptibility to vibriosis and that there is a significant interaction of this seasonal effect with ploidy. However, no significant advantage of triploidy was observed. Triploid oysters even showed lower survival than diploid counterparts in winter. Results are discussed in relation to differing energy allocation patterns between diploid and triploid Pacific oysters.     

Phenotypic and genetic consequences of size selection at the larval stage in the Pacific oyster (Crassostrea gigas)

The life histories of oysters in the genus Crassostrea, like those of most marine bivalves, are typified by high fecundity and low survival in nature. Rearing conditions in hatcheries however ensure optimized density, diet, and temperature. Hatcheries are becoming increasingly important for the production of juveniles in aquaculture, and their culture practices often include culling of slow growing larvae to reduce and synchronize the time taken to reach settlement. Because previous studies have found substantial genetic variation for early life developmental traits in Crassostrea gigas, these culling practices are likely to cause highly different selective pressures in hatcheries from those in the natural environment. We studied the phenotypic and genetic impact of such culling practices in a factorial cross between 10 males and 3 females subjected to progressive culling of the smallest 50% of larvae, compared with a non-culled control. Measurements were made on larval growth, survival, time taken to attain pediveliger stage and settlement success. Culling had a larger effect on the variance of these larval traits than on their means. The larvae in culled cultures were approximately 10% larger than those in controls, whereas the coefficient of variation was reduced by 30-40%. Culling also reduced the mean time to settlement by 12% and its variance by 55%. Using a multiplexed set of microsatellite markers to trace parentage, we also estimated the variance in reproductive success in a controlled experiment to quantify the consequences of intensive hatchery rearing practices. We also focused on changes in effective population size and genetic structure over time (and developmental stages). Our results show a loss of genetic diversity following removal of the smallest larvae by culling, as well as temporally varying genetic structure of the larval population. This supports the existence of genetic variability in early life developmental traits in C. gigas. Culling in hatcheries, like size-related selective pressures in the wild, are likely to have a significant genetic impact, through their effects on the timing of settlement.