Broodstock conditioning of the Portuguese oyster (Crassostrea angulata,Lamarck, 1819): influence of different diets

The Portuguese oyster Crassostrea angulata shows great potential in oyster farming. The conservation of pure populations of this species is important for production diversification and biodiversity preservation. In this way, the zootechnological development for seed hatchery production is extremely important. Broodstock conditioning is a key step in the process of rearing bivalves in a hatchery. Many factors regulate the reproductive cycle, being food one of the most important ones. To evaluate the effect of different diets on C. angulata reproductive performance, broodstock were conditioned with different food regimes formulated fundamentally by flagellates (Diet 1 – Pavlova lutheri and Isochrysis galbana clone T‐ISO; Diet 2 – P. lutheri, T‐ISO and Skeletonema costatum) and constituted fundamentally by diatoms (Diet 3 – S. costatum and Chaetoceros calcitrans; Diet 4 – P. lutheri, S. costatum and C. calcitrans). During conditioning, samples of oysters were collected to evaluate condition index, gonadal development and biochemical composition. At the end of the conditioning period, oysters were induced to spawn to evaluate reproductive output (fecundity, fertilization rate and D‐larvae development). The diets had an impact on the gametogenesis process, energy storage and reproductive output performance, being the best results those obtained in broodstock fed with the diatoms‐predominant diets. However, those fed with diets majority flagellates had an unsuccessful performance. Holistic approaches incorporating all results in this study reveal and reinforce the idea that the diatom species used presented the nutritional requirements to C. angulata broodstock, being essential in the conditioning phase.     

A comparative study of the survival,growth and gonad development of the diploid and triploid Hong Kong oyster,Crassostrea hongkongensis (Lam & Morton 2003)

The Hong Kong oyster, Crassostrea hongkongensis is an economically important and widely cultivated oyster species in southern China. Aiming to improve the aquaculture potentials of C. hongkongensis, a triploid organism was generated by inhibiting the second polar body formation in the fertilized eggs, using 0.5 mg L^?1 cytochalasin B. Triploids exhibited significantly higher growth and were larger than the diploids throughout the life cycle. Much of the triploid growth advantage can be attributed to resource reallocation to structural development due to gonadal sterility. Most of the triploid oysters exhibited polymorphic sterility with atrophic gonads and abnormal gametogenesis. Only few ‘dysfunctional’ sperms were produced by the triploid males, thus ruling out reproductive and genetic contamination of natural oyster populations from triploid cultivations. This advantage in addition to the superior growth of triploids makes it a potent alternative in Hong Kong oyster aquaculture industry. Triploid survival was mostly found to be similar to that of diploids except for day 360, when a significant triploid advantage in terms of survival was noted. Furthermore, we found that growth and survival of Hong Kong oyster is significantly higher at the Zhuhai site in comparison to the Beihai site. Our results indicate a strong environmental effect in Hong Kong oyster cultivation and points to the fact that Zhuhai site was more suitable for oyster farming than the Beihai site. Overall, this study provides valuable information to the aquaculture industry of southern China.     

Survival and growth of reciprocal crosses between two stocks of the Hong Kong oyster Crassostrea hongkongensis (Lam & Morton, 2003) in southern China

The Hong Kong oyster, Crassostrea hongkongensis, is one of the most economically important and well‐known oyster species in southern China. To explore the possibility of improving the performance of the Hong Kong oyster, complete diallel crosses between two oyster stocks, the Zhuhai stock (Z: fast growth line F4) and the Maowei Sea stock (M: resistant line F3), were conducted using pooled gametes. Three replicates, each consisting of two pure stocks (ZZ and MM) and two reciprocal crosses (ZM and MZ), were successfully generated. High fertilization rate and hatching level were observed among all the experimental groups, suggesting that there was no sperm–egg recognition barrier between geographic populations. Reciprocal crosses had higher survival rate compared to the pure stock crosses and the rate of survival increased with progeny growth. Growth heterosis became obvious both in larval and adult stages, and was primarily influenced by the egg origin and mating strategy at the larval stage. Also, the phenotypic traits of all progeny differed amongst the culture sites, suggesting a significant environmental effect. The Zhuhai site was more suitable for oyster aquaculture than the Maowei Sea site. Thus, our results demonstrated that crossbreeding between Zhuhai and Maowei Sea stocks of Hong Kong oyster to produce superior heterosis represents a promising means to improve the fishery yield of this species in southern China.     

Reproductive cycle of the rock oyster,Striostrea prismatica (Gray, 1825) from two locations on the southern coast of Ecuador

The reproductive cycle of the rock oyster Striostrea prismatica was determined at two fishing areas, General Villamil and Ayangue (located on the southern coast of Ecuador), between May 2012 and April 2013. Monthly sampling campaigns were performed at both locations. The tissues were histologically examined to determine gonadal index (GI), oocyte development, follicular area coverage and sex ratio. Surface seawater temperature, salinity and chlorophyll a concentration were measured during samplings. Our results show a similar annual reproductive pattern at both locations. The GI reached maximum values during the summer. Oysters reached highest ripeness in January and February, while spawning occurred in February–March. Gametogenesis was linked to a consistently increasing follicular area (from 1.3 ± 0.7% to 85.8 ± 7.8%) and associated to surface seawater temperature. Spawning coincided with warm water temperature fluctuations and a seawater salinity decrease. No correlation was found with Chlorophyll a concentration. The sex ratio of sampled populations was 1:1, suggesting that oysters sizing more than 10 cm in shell length present a stable sex proportion in the population. The diameter of mature oocytes was significantly reduced (32.7%) during histologically preparations in comparison to fresh oocytes. Our study provides useful information of environmental factors that may control the observed gametogenesis and spawning activity of S. prismatica.     

长牡蛎壳金选育系F4与普通养殖群体摄食和呼吸的比较研究

为了从生理学角度探讨长牡蛎壳金选育系的快速生长机制,本研究以两种规格长牡蛎(Crassostrea gigas)第4代壳金选育系(简称金1和金2)和两种普通养殖群体(简称对照1和对照2)为材料,比较了不同温度(16℃、20℃、24℃、28℃、32℃)和盐度(15、20、25、30、35)条件下各实验组的滤水率(FR)和耗氧率(OCR)。结果显示:(1)摄食实验中,4个组长牡蛎的滤水率均随温度或盐度增加先上升后下降,在温度28℃时出现最大值;对照1的滤水率在盐度25时达到最大值,其他3组滤水率在盐度30时达到最大值;方差分析结果显示,温度、群体,盐度、群体对长牡蛎的滤水率均有显著性影响,长牡蛎壳金选育系的滤水率均显著大于普通养殖群体(P0.05)。(2)呼吸实验中,较大规格长牡蛎壳金选育系的耗氧率受温度或盐度变化的影响更小。方差分析结果显示,温度、群体,盐度、群体均对长牡蛎耗氧率有显著性影响。实验温度范围内,金1的耗氧率显著小于对照1,但金2的耗氧率显著大于对照2(P0.05)。实验盐度范围内,壳金选育系的耗氧率均显著大于普通群体(P0.05)。(3)同一壳色群体中,个体越大,单位软体部干重滤水率和耗氧率越低。研究表明,较大规格长牡蛎壳金选育系F4更能适应外部环境的变化,该选育系的快速生长可能是由其较高滤食行为导致的。     

Rearing equipment on the quality aspects and growth performance of Crassostrea gigas (Thunberg, 1793): Implications for marketability

Shell characteristics and consequent marketability of the oyster Crassostrea gigas are influenced by several factors, and most important of which are the rearing methods and gears. In this study, we investigated the effects of two different gears on the quality aspect (shell shape, weight meat percentage, backward shell shape, epibionts and blisters) and growth rate (weight and length) of C. gigas by adopting the innovative Ortac Oyster Farming System, during the field grow‐out phase of production. Aspect and growth of oysters farmed in Ortac and floating bags were compared. After 14 weeks, the aspect of the animals was influenced by the gear, resulting in a lower quality in the Ortac (i.e. worse shell shape, lower weight meat percentage, higher percentage of backwards and blisters) than in the floating bag, while the incidence of epibionts was less in the Ortac. Regarding the growth performance, total wet weight gained in the Ortac was lower (~28.3 g) than in the floating bag (~32.4 g; p < .05). These results highlight the efficiency of the floating bag as oyster rearing system in the Mediterranean basin and suggest the need to alter the Ortac gear to these environmental conditions.     

Effect of different dietary microalgae combinations on growth and survival of black‐lip pearl oyster (Pinctada margaritifera) larvae and the feasibility of replacing microalgae with a dietary lipid emulsion

A 10‐day experiment was performed to examine different mono, binary and ternary dietary combinations on survival and growth of D‐shaped and umbone black‐lip pearl oyster, Pinctada margaritifera, larvae. The three tropical microalgae species were the flagellate Isochrysis galbana clone T. Iso (CS‐177) and diatoms Chaetoceros calcitrans (CS‐178) and Chaetoceros muelleri (CS‐176) which were fed to D‐shaped and umbone larvae at a density of 7000 and 14 000 cells mL^?1, respectively. A second experiment was performed to investigate the feasibility of replacing T. Iso with a lipid emulsion for both D‐shaped and umbone larvae for 10 and 12 days, respectively. The treatments included only T. Iso, unfed and lipid emulsion to substitute T. Iso at levels of 10% (LIP10), 30% (LIP30) and 100% (LIP100). In the first experiment, results showed that a monospecific diet of T. Iso led to significantly higher (P < 0.05) survival and growth of D‐shaped larvae than all the other treatments. Meanwhile, D‐shaped larval survival was significantly lower when only fed C. calcitrans as well as growth for those fed C. calcitrans or in combination with C. muelleri. However, for umbone larvae, survival and growth were significantly higher when fed a binary combination of T. Iso and C. muelleri or the ternary combination of T. Iso, C. muelleri and C. calcitrans compared with all other treatments. For the second experiment, results showed that with increasing lipid emulsion replacement, survival of both D‐shaped and umbone larvae significantly decreased (P < 0.05); however, the LIP100 treatment was not significantly different (P > 0.05) from the unfed treatment. For D‐shaped larvae, no significant growth difference was detected (P > 0.05) between the T. Iso and LIP10 fed treatments while for umbone larvae, the T. Iso, LIP10 and LIP30 were not significantly different (P > 005). These results indicate that microalgae combinations appear more necessary for later staged P. margaritifera larvae. In addition, the use of a lipid emulsion appeared to provide some nutrition to the larvae, although more research should be conducted to improve the use of such replacements.     

Establishment of a continuous cell line from female half‐smooth tongue sole (Cynoglossus semilaevis)

A new cell line (TSHC) derived from heart tissues was established from female half‐smooth tongue sole (Cynoglossus semilaevis), an economically important marine fish species in China. The cell line had been subcultured for more than 30 times over a period of 200 days. The cell line was optimally maintained at 24°C in minimum essential medium (MEM) medium containing foetal bovine serum (FBS), 2‐mercaptoethanol (2‐Me), sodium pyruvate, basic fibroblast growth factor (bFGF) and antibiotics. The TSHC cells were mostly composed of fibroblast‐like cells. Chromosome analysis revealed that the TSHC cell line had a normal diploid karyotype with 2n = 42, containing the heterogametic W chromosome. The TSHC cell line was susceptible to infection by flounder Lymphocystis disease virus (LCDV). Although an atypical cytopathic effect and only few of virus particles in the cytoplasm was observed, it provides a research material on the cell–pathogen interaction research about the viral infection of non‐host species.     

Development and characterization of a fibroblastic‐like cell line from caudal fin of the red‐line torpedo,Puntius denisonii (Day) (Teleostei: Cyprinidae)

A fibroblastic‐like cell line was established from the ornamental fish, red‐line torpedo (Puntius denisonii). The red‐line torpedo fin (RTF) cell line is being maintained in Leibovitz's L‐15 medium supplemented with 10% fetal bovine serum (FBS) for over 1 year at 28 °C on a continuous basis in normal atmosphere. The growth rate of RTF cells increased as the FBS proportion increased from 5% to 20% at 28 °C with optimum growth at the concentrations of 10% FBS. The morphology of RTF cell was predominantly fibroblastic like. Propagation of these cell lines was serum dependent, with a low plating efficiency (<15%). Karyotyping analysis of RTF cells at the 25th passage indicated that the modal chromosome number was 2n=50. The cell line was cryopreserved in liquid nitrogen at ?196 °C and could be recovered from storage after 6 months with good cell viability. Polymerase chain reaction amplification of a fragment of two mitochondrial genes, 16S rRNA and CO1, confirmed the identity of these cell lines with those reported from this animal species, confirming that the cell lines originated from P. denisonii. The bacterial extracellular products from Vibrio cholerae MTCC3904 and Aeromonas hydrophila were found to be toxic to RTF. The cell lines were not susceptible to viral nervous necrosis virus, a marine fish virus.     

Morphological aspects and histological effects of the attachment organ of Parabrachiella sp. (Copepoda: Lernaeopodidae) on the grey mullet,Mugil liza Valenciennes

The genus Parabrachiella Wilson, 1915 (Lernaeopodidae) is represented by copepods that are highly adapted to a parasitic way of life. In Argentina, only P. insidiosa var. lageniformis Heller, 1865, P. chevreuxii Van Beneden, 1891 and P. spinicephala Ringuelet, 1945 have been cited, but none of these have been reported on mugilids. Recently, other species of this genus were found attached to the nasal cavities of juvenile grey mullets, Mugil liza Valenciennes, from Samborombón bay, Buenos Aires province. In this study, the prevalence and mean intensity of the Parabrachiella sp. on grey mullet is investigated. In addition, the damage the parasite imposes on its hosts is examined through evaluation of histological sections and immunostaining for proliferative cell nuclear antigen (PCNA). The morphology of the parasite's bulla is described from light and scanning electron micrographs.     

壳白长牡蛎基因型与环境互作(G×E)效应分析

为探索壳白长牡蛎品系的壳色性状和生长性状的基因型与环境互作(G×E)效应,利用巢氏设计构建全同胞家系,每个家系分成两组分别在乳山和荣成海域进行养殖。利用线性混合模型和REML法分析11月龄壳白长牡蛎生长性状和壳色性状的遗传力及G×E效应。采用最佳线性无偏预测法(BLUP法)估计壳高和L~*两个性状的育种值,并通过加权获得综合育种值来筛选优良家系。结果显示,乳山组和荣成组的壳白长牡蛎生长和壳色性状的遗传力不同,分别为(0.14±0.08)～(0.62±0.18)和(0.01±0.03)～(0.78±0.19),可能存在尺度效应。以不同环境为固定效应,综合两个环境计算出的生长和壳色性状的遗传力为(0.02±0.02)～(0.51±0.09),然而由于部分全同胞家系缺失和模型不收敛的原因,估计模型中未包括母本/共同环境效应和显性效应,上述遗传力估计值偏高。本研究中生长和壳色性状在两个环境间的遗传相关为(–0.47±0.40)～(0.75±0.18),均小于0.8,表明壳白长牡蛎品系的生长和壳色性状都具有明显的重排效应,壳白长牡蛎品系其选育需要针对不同的养殖环境培育不同适应性的选育家系。综合育种值排名前20的个体其家系来源比例表明,家系G1和G21对于乳山海域表现出特殊的适应性,而家系G4、G22和G5对荣成海域环境具有特适性,家系G2则对两个环境具有普适性。研究为壳白长牡蛎品系的良种选育提供了重要的参考依据。     

Growth variations within a farm of mussel (Mytilus galloprovincialis) held near fish cages: importance for the implementation of integrated aquaculture

Fish farming releases extensive amounts of particulate organic waste that can be exploited by bivalves in integrated culture. We tested if mussels Mytilus galloprovincialis cultured at two depths (1 and 6 m) in a raft, moored 170 m from a fish farm had greater growth than bivalves held 550 m from the fish cages. Mussel growth was monitored monthly, covering the second phase of the culture, from thinning‐out to harvest (March to November 2011). We also studied if fish solid and dissolved nutrients increased the organic content of the seston and chlorophyll‐a levels near the fish cages through weekly samples. Results showed no differences in seston, chlorophyll and physiochemical characteristics of the water among rafts. Maximum growth and Condition Index (CI) occurred during spring–summer (April–August), when mussels had access to greater food quality and quantity. Mussels cultivated close to the cages showed similar shell length, weight and CI compared with mussels distant from the fish farm. Average shell length, meat dry weight and CI at harvest were 76.31 mm, 2.51 g and 23%. Bivalves cultured distant from the fish cages displayed 26% higher biomass than the other raft at the end of the experiment. Differences in biomass were explained by the significantly higher recruitment of mussel seed observed at the raft distant from the fish cages from June to November. The lack of a significant enhancement in growth of the bivalves cultured next to finfish is discussed.