Gonadal cycle of the dwarf oyster Ostreola stentina from the south of the Gulf of Hammamet on the eastern coast of Tunisia

The reproductive cycle of the dwarf oyster Ostreola stentina (Payraudeau 1826) at Monastir in the south of the Gulf of Hammamet on the east coast of Tunisia was studied using condition indices and histological techniques. The species was shown to be a protandric hermaphrodite. Whereas males dominated the small shell length categories (12.45–34.99 mm), females (21.00–47.16 mm) and hermaphroditic individuals (30.00–57.95 mm) become more predominant with increasing size, suggesting that individuals may undergo protandric development. The overall male to female ratio was 1.0:1.7. Gametogenic development begins in February–March for females and in September for males, and spawning occurs during April and July for females and December and July for males. There was a clear relationship between the gonadal condition index and gametogenic stage for both sexes. Values increased from zero in their early stage of development and peaked during ripe and spawning stages, before decreasing in the spent stage. However, the meat condition index did not show any statistical differences between gametogenic stages.     

长牡蛎(Crassostrea gigas)黑色壳表面微生物多样性的研究

为探究自然海区与室内养殖环境对软体动物壳色及壳表面细菌群落多样性的影响，以黑色壳长牡蛎（Crassostrea gigas）为研究对象，在自然海区与室内环境分别暂养30 d后，对长牡蛎的壳色及壳表面细菌群落的多样性和差异细菌功能进行分析。结果表明，室内养殖的黑壳长牡蛎出现较为明显的壳褪色现象，其黑色素含量显著下降，远低于自然海区养殖的黑壳长牡蛎。16S rRNA基因测序结果显示，变形杆菌门（Proteobacteria）、拟杆菌门（Bacteroidetes）、蓝细菌（Cyanobacteria）为长牡蛎壳表面的主要优势菌，其中变形杆菌的丰度超过50%。在门水平上，放线菌（Actinobacteria）、俭菌超门（Parcubateria）和Bacterial rice cluster 1（BRC1）在室内、外养殖长牡蛎之间存在较为显著的差异（P<0.05），而具有降解作用的放线菌和俭菌超门细菌在室内养殖的黑壳长牡蛎壳表面丰度较高。在属水平上，两组之间存在显著差异的细菌共计67种。该研究揭示了不同生境条件下，黑壳长牡蛎壳表面的微生物群落物种组成及差异，为解析长牡蛎壳色的形成机制提供了新的分析视角。     

Genetic variability of the Lessepsian migrant mussel Brachidontes pharaonis (Bivalvia: Mytilidae) in Tunisia

The present study used two mitochondrial markers (16S rRNA and COI) to assess the genetic diversity of a newly founded Lessepsian migrant mussel, Brachidontes pharaonis, in Tunisian waters. The species appears to be restricted to only one population in Rades Harbour, in the northern part of the country. Phylogenetic analyses revealed the monophyly of B. pharaonis in Tunisia. Both molecular markers revealed high genetic variability of the B. pharaonis population. Haplotype networks and demographic analyses confirmed the recent expansion events within this population. Multiple human-mediated introduction events involving several founder populations and intensive population growth rates are probably the main causes of the high polymorphism observed within this invasive mollusc.     

长牡蛎二倍体与三倍体壳型性状对体质量的影响分析

为了研究长牡蛎(Crassostrea gigas)二倍体和三倍体壳体性状对体质量的影响,随机选取养殖20月龄的二倍体和三倍体个体各75个,分别测量它们的壳体性状(壳高、壳长和壳宽)和体质量(全湿重和软体重),并进行多元回归和通径分析。结果显示,二倍体和三倍体的软体重变异系数都最高,绝大多数性状间的相关性达到了显著水平(P<0.05),全湿重和软体重的相关性较高分别为0.866和0.675;多元回归分析结果显示,壳长、壳高和壳宽对二倍体长牡蛎全湿重的影响相当,其中壳高最大,直接作用系数为0.474,对软体重影响最大的是全湿重,直接作用系数高达0.780;壳宽对三倍体长牡蛎全湿重的影响最大,直接作用系数为0.471,壳高和湿重对软体重的直接作用系数分别为0.348和0.367;对长牡蛎二倍体和三倍体全湿重和软体重分别进行回归,结果显示其r²值均小于0.85,说明存在对质量性状影响较大的其他因素。本研究为不同倍性长牡蛎的比较研究提供了数据支持。     

10个菲律宾蛤仔野生群体的遗传多样性研究

对我国沿海地区10个菲律宾蛤仔野生群体线粒体16S r RNA和COI基因部分序列进行测序,分别得到了长度为473bp和632bp的片段。结果表明,16S r RNA 193条序列A+T平均含量为66.6%,共检测到21个变异位点,193个个体具有22种单倍型;COI基因183条序列A+T平均含量为64.8%,共检测到126个变异位点,183个个体具有67种单倍型。基于群体间遗传距离利用Mega5.1软件构建10个群体的NJ树,聚类结果表明,大连群体和荣成群体聚为一支,其余8个群体聚为一支。AMOVA分析表明,大连群体和荣成群体间分化不显著,而荣成、大连群体与其余8个群体间的分化达到极显著水平(P0.01),说明我国沿海的菲律宾蛤仔野生群体存在一定的遗传分化。     

Significant genetic differentiation among meroplanktonic barrel jellyfish Rhizostoma pulmo (Cnidaria: Scyphozoa) in the Mediterranean Sea

Molecular data have shown that jellyfishes are more geographically restricted and evolutionarily divergent than previously thought. We examined genetic variation and divergence within the meroplanktonic barrel jellyfish Rhizostoma pulmo in the Mediterranean Sea; specific sampling areas were the northern Adriatic, western Mediterranean and Tunisian coast. A total of 19 sampling sites and 68 sequences of the mtDNA cytochrome c oxidase subunit I (COI) gene were used. Of the 68 COI sequences, 45 were newly collected specimens which originated from nine sampling sites along the Tunisian coast. A total of 24 haplotypes were obtained and the specimens sampled were characterised by relatively high levels of haplotype diversity (h = 0.866) and low levels of nucleotide diversity (π = 0.004). Haplotype network analysis showed the presence of three distinct phylogenetic lineages (populations), with separate geographic ranges in the northern Adriatic, western Mediterranean and Tunisian coast. The observed genetic differentiation between these three lineages was supported by the presence of significant genetic differentiation between the 19 populations (F_ST = 0.757, p < 0.001). The high level of genetic differentiation detected in the barrel jellyfish investigated could be attributed to either intrinsic and/or extrinsic barriers to genetic exchange between different populations that may have adapted to different environmental conditions.     

Source regions for recruitment of Calanus finmarchicus to Georges Bank: evidence from molecular population genetic analysis of mtDNA

Molecular population genetic analysis has provided evidence that the copepod, Calanus finmarchicus, of the Labrador Current, Gulf of St Lawrence, Scotian Shelf, Gulf of Maine, and Georges Bank constitute a single, interbreeding population. The DNA sequence of a 350 base pair portion of the mitochondrial large subunit (16S) ribosomal RNA (rRNA) gene was determined for a total of 72 individuals collected in 1992, and 110 individuals collected in 1993 from these regions. There was significant heterogeneity in haplotype frequencies among the samples collected in 1992, but this heterogeneity did not resolve into regional patterns. The only regional differences seen were between pooled samples of the western N. Atlantic and those of the Norwegian Sea. There were no significant differences in haplotype frequencies among the samples collected in 1993, and fewer haplotypes were observed in these samples. Intraspecific molecular variation was typical of other marine species: there were 29 haplotypes among the 182 individuals sequenced. The frequency distribution of the haplotypes was highly skewed: 128 individuals shared one haplotype and 19 individuals were unique. There were 24 variable sites among the 350 bases sequenced; estimated nucleotide diversity was 0.0042. The genetic character of C. finmarchicus populations in the western N. Atlantic was stable over time in that three of the haplotypes (including the most abundant) occurred in both 1992 and 1993. However, haplotype frequencies differed significantly between the two years. The lack of regional structure in the 1992 samples and the genetic homogeneity of samples collected in 1993 across the domain from the Labrador Current to the Gulf of St Lawrence to Georges Bank and the Gulf of Maine indicated that there is significant gene flow across this region. The persistent genetic pattern suggests that the Gulf of St Lawrence may be an important source region for recruitment of C. finmarchicus to Georges Bank. Determination of zooplankton dispersal patterns within high gene flow species will provide information that may not be determined by conventional oceanographic analyses.     

单体牡蛎诱导变态的研究

为了得到壳型规则、大小均一的单体牡蛎,作者以葡萄牙牡蛎(Crassostrea angulata)和长牡蛎(Crassostrea gigas)眼点幼虫为材料,从肾上腺素(EPI)浓度梯度、处理时间梯度和眼点幼虫密度梯度3个因素诱导产生不固着变态的单体牡蛎。结果表明,葡萄牙牡蛎用EPI处理24 h的最适浓度为5×10–4 mol/L,不固着变态率为72.8%,在该最适浓度下,最佳处理时间为12 h,不固着变态率为82.7%。长牡蛎用EPI处理6 h的最适浓度为5×10–5 mol/L,不固着变态率为53.2%,在该最适浓度下,最佳处理时间为8h,不固着变态率为56.8%,眼点幼虫密度在80个/m L以下EPI处理效果没有显著性差异。对长牡蛎幼虫进行后续生长测定,结果显示EPI处理组幼虫的壳长、壳高和存活率要高于对照组,表明EPI可能促进牡蛎幼虫变态长出次生壳并提高其生存能力。     

Differential recruitment of introduced Pacific oysters and native mussels at the North Sea coast: coexistence possible?

Pacific oysters (Crassostrea gigas Thunberg 1793) have been introduced into the Wadden Sea (North Sea), where they settle on native mussel beds (Mytilus edulis L.), which represent the only extensive insular hard substrata in this soft-sediment environment. As abundances of C. gigas rose, some mussel beds became increasingly overgrown with oysters, whereas others did not. Field experiments revealed that recruitment of C. gigas was higher in the lower intertidal than in the upper subtidal zone, that it was higher on conspecifics than on mussels, and that it was not affected by barnacle epigrowth except when settling on mussels. Mussel recruitment is known from inter- and subtidal zones. It occurred equally on oyster and mussel shells but showed a clear preference for barnacle epigrowth over clean shells. Assuming that settlement and recruitment are key processes for species abundances on the North Sea coast, it is predicted that the positive feedback in oyster settlement will lead to rapid reef formation of this invader at the expense of mussel beds. Mussels, however, may escape competitive exclusion by settling between or on the larger oysters especially when barnacles are abundant. Experimental patches with mussels were more often covered by fucoid algae (Fucus vesiculosus forma mytili Nienburg) than patches with oysters, and oyster recruitment was poor underneath such algal canopies. Thus, fucoids may provide the native mussels with a refuge from the invading oysters and the two bivalves may coexist, provided food is not limiting.     

基于线粒体16S rRNA基因序列对3种珍珠贝的鉴定

传统的形态鉴定主要依赖于分类者的经验和水平,容易产生分类错误。为提高物种鉴定的准确性,作者采用形态学与线粒体16S rRNA基因序列分析相结合的方式对广西北海涠洲岛采获的3种珍珠贝进行了鉴定。研究结果表明,基于形态学可将3种珍珠贝分别鉴定为企鹅珍珠贝(Pteria penguin)、宽珍珠贝(P.loveni)和中国珍珠贝(P.chinensis),但基于16S r RNA基因序列,形态学鉴定为P.loveni的标本应为P.lata。P.loveni和P.lata是两个完全独立的种,并非为同物异名。珍珠贝属16S rRNA基因序列的种间遗传距离明显大于种内遗传距离,适宜作为该属种类鉴定的分子标记。在基于16S r RNA基因序列构建的系统树中,相同种类的不同个体均形成单系群,并获得很高的支持率。分布于中国沿海的珍珠贝属种类应包括P.lata,是否有P.loveni的分布尚需进一步证实。     

Modelling growth and reproduction of the Pacific oyster Crassostrea gigas: Advances in the oyster-DEB model through application to a coastal pond

A bio-energetic model, based on the DEB theory exists for the Pacific oyster Crassostrea gigas. Pouvreau et al. [Pouvreau, S., Bourles, Y., Lefebvre, S., Gangnery, A., Alunno-Bruscia, M., 2006. Application of a dynamic energy budget model to the Pacific oyster, C. gigas, reared under various environmental conditions. J. Sea Res. 56, 156–167.] successfully applied this model to oysters reared in three environments with no tide and low turbidity, using chlorophyll a concentration as food quantifier. However, the robustness of the oyster-DEB model needs to be validated in varying environments where different food quantifiers reflect the food available for oysters, as is the case in estuaries and most coastal ecosystems. We therefore tested the oyster-DEB model on C. gigas reared in an Atlantic coastal pond from January 2006 to January 2007. The model relies on two forcing variables: seawater temperature and food density monitored through various food quantifiers. Based on the high temperature range measured in this oyster pond (3–30 °C), new boundary values of the temperature tolerance range were estimated both for ingestion and respiration rates. Several food quantifiers were then tested to select the most suitable for explaining the observed growth and reproduction of C. gigas reared in an oyster pond. These were: particulate organic matter and carbon, chlorophyll a concentration and phytoplankton enumeration (expressed in cell number per litre or in cumulative cell biovolume). We conclude that when phytoplankton enumeration was used as food quantifier, the new version of oyster-DEB model presented here reproduced the growth and reproduction of C. gigas very accurately. The next step will be to validate the model under contrasting coastal environmental conditions so as to confirm the accuracy of phytoplankton enumeration as a way of representing the available food that sustains oyster growth.     

基于线粒体COI和16S rRNA基因研究3个地理群体黑龙江河蓝蛤的遗传多样性

采用通用引物对辽宁盘锦、辽宁大连、山东日照的3个地理群体黑龙江河蓝蛤(Potamocorbula amurensis)COI和16S r RNA序列进行扩增、测序分析,得到30条658bp的COI基因部分序列和27条450 bp的16S r RNA基因部分序列。其中COI和16S r RNA基因部分序列T、C、A、G和A+T的平均含量分别为45.4%和32.0%、13.5%和13.3%、20.7%和29.3%、20.4%和25.3%、66.1%和61.3%,AT含量高于GC含量,这与其他软体动物门动物的COI和16S r RNA的观测结果相近。COI和16S r RNA分别检测到了24个单倍型、43个核苷酸多态位点和9个单倍型、19个核苷酸多态性位点。AMOVA分析表明,3个群体间COI和16S r RNA部分基因总遗传分化系数分别为Fst=0.0090(P0.001)和Fst=0.0674(P0.001),群体内遗传分化远大于群体间、群体内存在较高的遗传分化。用NJ法构建分子进化树,3个地理群体的黑龙江河蓝蛤聚为一个族群,有少数个体和其他群体的个体聚在一起。     

利用排水称重法测定、分析不同海区牡蛎的贝壳密度

不规则物体体积测量的传统方法有排液法、称重法和排气法等,不仅操作繁琐,而且准确度及精确度低。本文首次提出了排水称重法,利用该方法能够更加简单、快速、准确地测量出物体的体积和质量。用此法对不同海区26个采样点的1100个牡蛎贝壳密度进行了测定分析,结果表明:大部分牡蛎的左壳密度显著小于右壳密度(P0.05);对于不同海区间的牡蛎,贝壳密度存在极显著的差异(P0.001);分布海区相近的牡蛎间,壳的密度也趋于相近;受环境压力相对较大的潮间带上部的牡蛎,贝壳密度也相对较大。本研究为剖析牡蛎贝壳形成机制以及温度、盐度和海洋酸化等环境因素对牡蛎贝壳形成的影响等研究提供了实验数据和参考。     

大银鱼和小齿日本银鱼线粒体细胞色素b和16S rRNA基因部分序列分析

对大银鱼和小齿日本银鱼的线粒体细胞色素b和16S rRNA基因片段进行了扩增和序列测定，分析比较了2种间的序列差异。大银鱼2个个体间细胞色素b基因序列无差异，小齿日本银鱼3个个体的序列出现了2种单倍型；大银鱼同小齿日本银鱼细胞色素b序列(单倍型SB-1)之间存在86个碱基差异(差异率21％)，变异较大。大银鱼、小齿日本银鱼的16S rRNA基因片段种内个体间无序列差异，两种间存在21个碱基的差异(差异率5％)，有1个碱基的插入／缺失。由此可见，2种银鱼间线粒体细胞色素b基因的进化速率较快，约为16S rRNA基因片段的4倍。根据细胞色素b序列数据。推算出2种银鱼大概在中新世晚期发生分化。     

Polybrominated diphenyl ethers and their methoxylated analogs in mullet (Mugil cephalus) and sea bass (Dicentrarchus labrax) from Bizerte Lagoon, Tunisia

Concentrations of ten polybrominated diphenyl ethers (PBDEs) and eight methoxylated polybrominated diphenyl ethers (MeO-PBDEs) in mullet (Mugil cephalus) and sea bass (Dicentrarchus labrax) collected from the Bizerte Lagoon and the Mediterranean Sea were investigated. To the best of our knowledge, this is the first report of these compounds in marine fishes from Tunisia. The PBDE mean concentrations in fish from Bizerte Lagoon were 45.3 and 96.2 ng g(-1) lw respectively in mullet and sea bass, while the concentrations of these compounds in mullet and sea bass from Mediterranean Sea were 7.80 and 27.9 ng g(-1) lw respectively. MeO-PBDE concentrations in mullet and sea bass from Bizerte Lagoon ranged from 6.46 to 286 ng g(-1) lw and from 49.4 to 798 ng g(-1) lw respectively, while the concentrations of these compounds in mullet and sea bass from Mediterranean Sea ranged from 190 to 401 ng g(-1) lw and from 353 to 578 ng g(-1) lw respectively. The total PBDEs and total MeO-PBDEs concentration in fish from Bizerte Lagoon were similar or slightly lower than those reported for other species from other locations around the world.     

Reductions in TBT concentrations in UK estuaries following legislation in 1986 and 1987

In 1986 the Government of the United Kingdom (UK) introduced legislation to control the sale of tribytltin (TBT)-based paints. From 1986 to 1989 monitoring was undertaken, whereby samples of water, sediment, oysters (Crassostrea gigas) and mussels (Mytilus edulis) were collected from six estuaries and water samples from five marinas/harbours, and analysed for TBT. In 1989 the concentrations of TBT in water, oysters and mussels were generally only one-third to one-quarter of those observed in 1986. The most marked decrease in concentrations of TBT in oysters and mussels occurred in 1987–1988, following the 1987 ban on the use of TBT-based paints on small boats and mariculture equipment. As the concentrations of TBT in oyster tissues had decreased there has been improvement in oyster growth in terms of both meat production and growth of thin (i.e. normally shaped) shells. Oysters with a normal shell shape can now be grown in five of the six estuaries which were monitored. In contrast to the reduction in concentrations of TBT noted in water and bivalves, changes in concentrations of TBT in sediments showed no clear trend from 1986 to 1989.     

马氏珠母贝黄壳色家系的生长、壳色分离及总抗氧化能力比较

以马氏珠母贝(Pinctada fucata martensii)黄壳色作为选育指标构建了黄壳家系,对其4个生长性状和黄壳色性状分离比进行了统计分析,并测定了黄壳家系中黄壳、褐壳色个体不同组织中的总抗氧化能力(TAC)。结果发现:黄壳色性状能遗传并发生分离,其分离不完全符合孟德尔遗传分离定律;褐壳色亲本不会产生黄壳色子代,推测褐壳色是纯合子;第一代黄壳家系5、7月龄贝仅壳宽值显著小于褐壳家系(P0.05),壳高、壳长、总质量差异不显著, 11月龄贝仅壳长和壳宽值显著小于褐壳家系, 14、20月龄贝的壳高、壳长、壳宽、总质量这4个生长性状均极显著小于褐壳家系(P0.01);在第一代6个黄壳家系内,黄壳、褐壳色个体的4个生长性状差异不显著;黄壳色贝在足、心、性腺中的TAC显著低于褐壳色贝(P0.05);在鳃、闭壳肌、消化腺组织中TAC高于褐壳色贝,但差异不显著。     

脊尾白虾(Exopalaemon carinicauda)3 个野生群体mtDNA 16S rRNA 序列差异及长臂虾科系统进化关系

采用PCR扩增和序列测定等技术,对脊尾白虾3个野生群体共计58个个体的线粒体16SrRNA基因片段进行初步研究。结果表明,在获得的长度为509bp核苷酸序列中,共检测到7个变异位点、8种单倍型,除象山湾群体外其它两个群体遗传多样性水平都较低。AMOVA分析表明,象山湾与莱州湾群体有显著的遗传差异,其余群体间的遗传差异不显著。另外,将本研究所得序列与GenBank中长臂虾科11个种的16SrRNA基因片段序列进行比较后,发现其聚类关系与传统分类略有不同,并依据16SrRNA序列变异百分比推测了长臂虾科12个种的大致分化时间。     

Comparison and history of Polydora websteri and P. haswelli (Polychaeta: Spionidae) as mud-blister worms in New Zealand shellfish

Shells of commercially valued bivalves in New Zealand, Crassostrea gigas, Perna canaliculus and Pecten novaezelandiae, are damaged by blister-causing Polydora polychaete species known to be close in morphology to the widely recorded oyster pest Polydora websteri Hartman. Recent New Zealand occurrences are here confirmed to relate to two species, P. websteri, and a second similar species, Polydora haswelli Blake & Kudenov, a new record for New Zealand, previously known only from Australia; the two species are described and compared. The worms have limited distributions, with P. websteri confirmed only for Pacific oysters (C. gigas) in northern New Zealand, although prior reports indicate it may also occur on scallops and have reached the northern South Island. Polydora haswelli has been found only in northern New Zealand, occurring on subtidal mussels and scallops and native oysters (Perna canaliculus, Pecten novaezelandiae, Ostrea chilensis), as well as co-existing with intertidal P. websteri on Pacific oysters. The worms are not present in Foveaux Strait O. chilensis beds, a major source of past oyster exports to Australia. The history of mud-blister worm outbreaks in Australasia is examined. While trans-Tasman exports of live oysters from New Zealand were commonplace during the nineteenth century, there is no evidence that mud-blister worms were present in New Zealand then. The earliest reports only date from the early 1970s and only from northern New Zealand, whereas a century earlier in the 1870s at least one of these pest worms had become widespread along eastern Australian coasts.     

Effects of epibionts on Sabellaria alveolata (L.) biogenic reefs and their associated fauna in the Bay of Mont Saint-Michel

Biogenic structures built by ecosystem engineers such as corals, bivalves, polychaetes, and sea grasses provide habitat for benthic vertebrates and invertebrates. The polychaete Sabellaria alveolata is an important foundation species whose reef structure adds topographic complexity and high levels of biodiversity to the otherwise low-relief, low diversity, soft-bottom environments in the Bay of Mont Saint-Michel, France, where the largest such reef formations in Europe are found. In this bay, reefs are being increasingly colonised by oysters (Crassostrea gigas) from local aquaculture operations and by green algae (Ulva sp.) due to the increasing inputs of nitrates from terrestrial origin. The purpose of this study was to investigate the possible impacts of epibiotic oysters and green algae on the S. alveolata population and reef community structure in the Bay of Mont Saint-Michel, France. Univariate and multivariate comparisons of macrofauna were conducted for five reef types: controls (no epibionts), low oyster density, high oyster density, green algae, and oyster and green algae. Results showed that all the three reef types with oysters had significantly higher species richness and diversity values than control and algae-only reef types. Pairwise ANOSIM and SIMPER comparisons of controls versus the four reef types with epibionts revealed that all three of the reef types with oysters were significantly different from controls, but there was no significant difference between controls and algae-only reef types. A striking feature of the reef comparisons is that no single species in this species-rich system contributed more than 8.86% to the dissimilarity between the reef types. Thus, k-dominance curves for species abundances were not effective in revealing differences among the reef types. Our results demonstrate that recent anthropogenic inputs of oysters affect the reef species assemblage more strongly than algal epibionts. In addition, epibionts, especially green algae, alter S. alveolata population structure, causing a reduction in new recruits that over the long run may cause significant damage to the reef structure itself. These results are a first step towards understanding anthropogenic threats to S. alveolata reefs and may be useful in the development of strategies for their protection and management.