Biochemical and fatty acid dynamics during larval development in the razor clam Ensis arcuatus (Bivalvia: Pharidae)

This article reports on the changes in gross biochemical and fatty acid composition of the razor clam Ensis arcuatus larvae throughout development. Protein was the largest component of dried larval tissues. The energy required for embryogenesis in E. arcuatus oocytes was obtained from stored proteins and carbohydrates, while total lipids increased significantly. Lipids and carbohydrates have an important role as energy contributors from day 1 to day 8. During larval development the strategy was to indistinctly store energy reserves (protein, lipid and carbohydrate) to surpass metamorphosis. During embryonic development there was a gain in fatty acids of neutral and polar lipids. A depletion of fatty acids in neutral lipids was observed from day 1 to day 8 in E. arcuatus larvae, mainly due to the decrease in 16:0 and EPA. NMID fatty acids were present in amounts similar to those of some PUFAs in polar lipids, especially 22:2NMID.     

Effects of stocking density on intermediate culture of the razor clam Ensis arcuatus (Pharidae: Bivalvia)

Commercial production of most bivalve species involves a phase of intermediate cultivation during which juveniles are grown under protected conditions until they can be transferred to the final grow‐out location. Consequently, the aim of this study was investigating the effect of density on growth and survival in the intermediate culture of the razor clam Ensis arcuatus in raft. Two series of experiments were performed, using in the first experiment two holding systems: 5‐L plastic bottles and PVC cylinders, both perforated, covered with a 1 mm mesh net to prevent razor clam escape and a 10‐cm layer of coarse grained sand (300–1200 μm grain diameter). In this trial, two stocking densities were tested: 0.15 and 0.30 kg m^?2. In the second one‐two densities were essayed (0.62 and 1.24 kg m^?2) in the PCV cylinders. In the first experiment after 27 of trial higher growth in cylinders at low density was observed. At the end of this trial all juveniles died in the 5‐L plastic bottles and higher growth and survival were recorded in the cylinders at low density. In the second experiment, significant differences in growth and survival were found on day 70, with the best results at low density. Five‐L plastic bottles must be rejected as holding system for the intermediate culture of E. arcuatus and the PVC cylinders at low densities could be a good alternative. At the end of the experiment razor clams reached the adequate size for their transfer into the grow‐out system.     

Genetic variation of the razor clam Ensis siliqua (Jeffreys, 1875) along the European coast based on random amplified polymorphic DNA markers

Ensis siliqua is regarded as an increasingly valuable fishery resource with potential for commercial aquaculture in many European countries. The genetic variation of this razor clam was analysed by randomly amplified polymorphic DNA (RAPD) in six populations from Spain, Portugal and Ireland. Out of the 40 primers tested, five were chosen to assess genetic variation. A total of 61 RAPD loci were developed ranging in size from 400 to 2000 bp. The percentages of polymorphic loci, the allele effective number and the genetic diversity were comparable among populations, and demonstrated a high level of genetic variability. The values of Nei's genetic distance were small among the Spanish and Portuguese populations (0.051–0.065), and high between these and the Irish populations. Cluster and principal coordinate analyses supported these findings. A mantel test performed between geographic and genetic distance matrices showed a significant correlation (r=0.84, P<0.05), suggesting an isolation by distance process.     

Identification of razor clams <Emphasis Type="Italic">Ensis arcuatus</Emphasis> and <Emphasis Type="Italic">Ensis siliqua</Emphasis> by PCR-RFLP analysis of ITS1 region

ABSTRACT:   Ensis arcuatus and Ensis siliqua are the most economically important species of razor clams in the European Union. Due to similarities between their shell morphology, and the differing retail value, these species are often misidentified. Therefore, it is necessary to develop an appropriate protocol to allow accurate differentiation between these species of razor clam in order to protect consumer rights, avoid commercial fraud (whether intentional or unintentional), and to enforce labeling and safety regulations. With the aim of developing a rapid and reliable method of differentiation, individuals of E. arcuatus and of E. siliqua were examined by polymerase chain reaction restriction fragment polymorphism (PCR–RFLP) using the internal transcribed spacer region 1 (ITS1). A species-specific restriction endonuclease pattern was found with the enzyme Ksp I for both species, allowing their exact identification. Thus, this work provides a simple, reliable and rapid protocol for accurate discrimination between E. arcuatus and E. siliqua , which proves useful for traceability and enabling the enforcement of labeling regulations.     

Larval and post‐larval growth,spat production and off‐bottom cultivation of the smooth venus clam Chionista fluctifraga

Harvesting practices of the clam Chionista fluctifraga show a decline in commercial size and densities, but no strategies have been developed to maintain clam beds. Aquaculture represents an alternative for preserving this resource. Adult clams from commercial grounds were used as broodstock. Conditioning, induction of spawning, cultivation of larvae, settlement of eyed larvae and nursing of postlarvae were performed in the hatchery for producing spat. Larvae and postlarvae were used to measure increase in shell height and data were fitted to exponential growth models. Spat were placed in floating trays and maintained in off‐bottom cultivation for 9 months. Samples of clams and tissues were collected monthly to measure absolute growth, shell height increase and a condition index. Larvae, postlarvae and juveniles showed exponential growth patterns. Mean shell height increased about 0.030 mm day^?1 during larval and post‐larval stages and 0.049 mm day^?1 during field cultivation. Pediveligers (height 215 ± 83 μm) entered metamorphosis at days 9–13 after fertilization, and postlarvae reached 3011.7 ± 325.5 μm (height) at day 60. After field cultivation, survival was about 95%; juvenile shell height was 20.6 ± 2.2 mm, and total weight was 5.3 ± 0.7 g. Growth rates were superior to natural conditions and the condition index was high throughout the study. Our results show that spat of C. fluctifraga can be produced in the hatchery, and that field production can be maintained in off‐bottom trays until reaching commercial size. Aquaculture activities for this species need to be established and evaluated.     

Vibrios in hatchery cultures of the razor clam,Solen marginatus (Pulteney)

Hatchery culture of the razor clam, Solen marginatus (Pulteney), has recently been developed in Galicia (NW Spain). However, recurrent episodes of mortalities of larval and post‐larval cultures have been recorded during the course of various studies. The disease signs were similar to those described for other bivalve species in outbreaks caused by bacteria of the genus Vibrio. In this article, we present the results of microbiological monitoring of two batches of razor clams with different survival rates. All fermentative isolates were identified as members of the Splendidus clade within the genus Vibrio. Some of these isolates, identified as Vibrio splendidus‐like, were clearly associated with the batch suffering mortalities, indicating their possible role as pathogens. Similar strains were found in the broodstock, suggesting vertical transmission of these bacteria. This is the first study of the microbiota associated with hatchery culture of S. marginatus, and the results will provide useful information for the optimization of a protocol for hatchery culture of this bivalve species.     

Larval rearing and spat production of edible oyster Crassostrea gryphoides (Schlotheim)

The edible oyster Crassostrea gryphoides was induced to spawn in the laboratory by thermal stimulation, and the larvae were reared to spat. The larvae grow through the straight hinge (70×60.5 μm), early umbo (96.25×87.5 μm), late umbo (215×152.5 μm), eyed stage (290×222.5 μm) and pediveliger (308.25×254.17 μm) in pelagic phase before metamorphosing to spat, and these stages are described. These stages were reached at 20 h, on the fifth, 11th, 15th, 19th and 21st day respectively. Spat settlement percentage was found to be 1.8%.     

鳞砗磲的人工繁育和早期发生

基于2015年的预备性试验,于2016年3—6月在三亚开展了鳞砗磲人工繁育技术研究。结果显示,采用五羟色胺催产剂可以有效促使鳞砗磲排放配子,精卵比例为50∶1~100∶1时受精率、孵化率较高,孵化密度控制在15~20个/m L较适宜,经过36 h孵化,获得初孵D形幼虫;优选的D形幼虫经过5 d的微充气培养,即发育至足面盘幼虫,进入附着变态期。利用净水采苗法促使幼虫完成变态,经过7~10 d,幼虫出现鳃、次生壳,并建立完善的虫黄藻系统,即完成变态、形成稚贝。利用一定浓度虫黄藻浸泡足面盘幼虫2 h以内,可以有效地提高变态率。采用微流水+微充气模式和2000~3000 lx光照(白天)进行稚贝培养,稚贝利用体内虫黄藻提供的营养就可以继续完成贝壳生长、器官发育。经过48 d和65 d的培育,分别成功获得壳长×壳高=(1174.0±146.5)μm×(1208.0±135.3)μm的稚贝约3万个,壳长×壳高=(1750.2±224.1)μm×(1816.5±226.5)μm的幼贝约0.5万个。本研究首次在国内人工繁育成功,获得了鳞砗磲苗种,为鳞砗磲人工大规模繁育、增殖放流、移植保育等工作奠定了基础。     

Environmental impact of razor clam harvesting using salt in Ria Formosa lagoon (Southern Portugal) and subsequent recovery of associated benthic communities

• 1. Razor clams are found in different habitats ranging from sheltered systems (e.g. lagoons and estuaries) to open coasts. They are distributed worldwide and comprise a small number of species of high economic value. Depending on the specific habitat and species exploited, different mechanical and hand‐harvesting techniques are employed. While the environmental effects resulting from mechanized methods have been described by several authors, the impacts caused by traditional hand‐harvesting methods remain unknown. Therefore, a study was undertaken in Ria Formosa lagoon (South of Portugal) addressing the environmental effects resulting from harvesting Solen marginatus with salt.
• 2. No significant impact on the sediment was found; the main effect was an increase in salinity after covering the area with salt, which decreased rapidly with the flood tide and after a few hours had returned to pre‐harvesting levels.
• 3. No effects on benthic communities were observed, with similar fluctuation patterns recorded in control and experimental areas, the observed differences being attributed to the natural variability of benthic populations.
• 4. Based on the results obtained in the present study, a razor clam fishery using salt in intertidal areas can be considered environmentally ‘friendly’.

Copyright © 2008 John Wiley & Sons, Ltd.     

Captive spawning and larval rearing of California yellowtail (Seriola lalandi)

A captive population of California yellowtail (Seriola lalandi) was used to document spawning patterns, including measures of egg production, population fecundity and egg and larval quality from 2007 to 2010. Spawned eggs were also used to document larval development and to develop rearing techniques for aquaculture in the region. Broodstock growth and condition factor were best when feeding rations were maintained at 10–15% body weight week^?1 during the warm summer months. A winter ration based on satiation feeding was typically 4% body weight week^?1. During the 4‐year study period, the only broodstock health issue was an infestation by the parasitic gill fluke Zeuxapta seriolae, which was readily treated. Spawning occurred naturally in the 140 m³ tank when the ambient water temperature reached 16°C and ended when the temperature exceeded 22°C. Egg production reached a maximum in 2010 when 43 spawn events were recorded from a pool of nine females yielding 36.8 million eggs in total. The average female size at this time was 20 kg, which equated to a total annual population fecundity of approximately 226 000 eggs kg^?1 female year^?1. Larval rearing trials yielded survival rates as high as 5.8% from egg to 50 days post‐hatch (dph). Successful larval culture methods included the addition of algae paste for green water culture, rotifers (20 rotifers mL^?1) at 2 dph and Artemia (5 Artemia mL^?1) at 6 dph. Larvae were transferred from the incubation tank at 10 dph to a shallower tank with 33% greater surface area to accommodate the larvae's strong orientation to surface waters. This research represents the first documentation of successful spawning and larval rearing for S. lalandi in the eastern Pacific.     

The larval rearing of the marine ornamental crab, Mithraculus forceps (A. Milne Edwards, 1875) (Decapoda: Brachyura: Majidae)

The goal of this study is to develop a larviculture protocol for Mithraculus forceps, a popular marine aquarium species. Different temperatures (25±0.5°C and 28±0.5°C), stocking densities (10, 20, 40 and 80 larvae L^?1), prey densities (newly hatched Artemia of 1, 4, 7 and 12 nauplii mL^?1) and metamorphosis to crab conditions (Systems A and B) were tested. The best survivorship and faster development were obtained when the larvae were reared at a density of 40 larvae L^?1 for 7 days post hatching (DPH) in System A, at 28°C and fed with 7 mL^?1 of newly hatched Artemia nauplii. After 7 DPH all the megalopa were moved to System B and the same temperature and prey density were maintained. At the end of the experiment, 12 DPH, survivorship of 74.1±4.8% was obtained.     

Effects of low dissolved oxygen on the digging behaviour and metabolism of the hard clam (Meretrix lusoria)

The dissolved oxygen (DO) concentration that induces the onset of anaerobic metabolism in hard clams was found to be 1.11 mg O₂ L^?1, at which time, the concentration of succinate in the body fluid was 4.4 μmol mL^?1. When the DO concentration was <1.11 mg O₂ L^?1, the burial depth was significantly reduced, and succinate significantly accumulated in the body fluid. After 24 h of anoxic exposure, succinate had accumulated in the gills and equal amounts of succinate and alanine had accumulated in the foot. This indicates that carbohydrates in the gills and amino acids in the foot contribute to anaerobic energy production. The accumulation rates of succinate and propionate in the body fluid were the highest compared with those in other tissues, while no accumulation of alanine in the body fluid was found. The recovery rates of succinate in the body fluid and alanine in the foot were the highest compared with those in other tissues. The results of this study suggest that the DO concentration in the bottom water of clam ponds should be maintained at ≥1.11 mg O₂ L^?1, and the anoxia‐tolerant ability of hard clams can be assessed by the contents of carbohydrates.     

Reproductive cycle,collection and early growth of Aulacomya ater,Molina 1782 (Bivalvia: Mytilidae) in northern Chile

We evaluated the reproductive cycle of Aulacomya ater at Caleta Punta Arenas (Antofagasta, Chile), finding two important spawning periods (August–September and October–November) associated with declining water temperatures. Mytilid collectors were installed at Caleta Punta Arenas and Caleta Errázuriz (250 km further south) in November and then extracted in consecutive time periods. The results showed settlements of A. ater along with Choromytilus chorus (Molina) and Semimytilus algosus (Gould). An analysis of the spat size structure from each period revealed different micro‐cohorts for each species; the micro‐cohorts of C. chorus were predominant. Daily growth rates were estimated using the average sizes of the micro‐cohorts on two consecutive dates. Although the daily growth rates did not vary significantly between sites for a given species, these rates were lowest for A. ater and highest for S. algosus. The abundance and greater growth of C. chorus caused interspecific competition with A. ater that was detrimental to the survival of the latter. We hypothesize the existence of a metapopulation of A. ater in northern Chile that would explain the settlement of four micro‐cohorts on collectors after 28 days at Caleta Punta Arenas, and the settlement recorded after 80 days at Caleta Errázuriz.     

Underestimation of primary productivity on continental shelves: evidence from maximum size of extant surfclam (Spisula solidissima) populations

Atlantic surfclams (Spisula solidissima), among the largest extant non‐symbiotic clam species in the world, live in dense aggregations along the Middle Atlantic Bight (MAB) continental shelf. The food resources that support these populations are poorly understood. An individual‐based model that simulates the growth of post‐settlement surfclams was used to investigate the quantity of food needed to maintain existing surfclam populations along the MAB continental shelf. Food inputs to the model were based on measured near‐bottom water‐column chlorophyll concentrations. Simulations showed that these water‐column food sources supported only 65% of the observed body mass of a standard large surfclam (160‐mm shell length). Additional simulations using benthic food sources to supplement water‐column food sources by 20% generated surfclams that grew to observed size and biomass and exhibited spawn timing consistent with the known surfclam spawning season. The simulation results suggest that measured water‐column chlorophyll concentrations may underestimate the food available to the continental shelf benthos. Large continental shelf bivalves are an essential resource for fisheries and higher trophic level consumers. Understanding available and utilized food resources is important for predicting long‐term impacts of climate change on benthic secondary production and fishery yield on the continental shelf.     

Abnormal mortalities of the carpet shell clam Ruditapes decussatus (Linnaeus 1756) in natural bed populations: a practical approach

In situ controlled systems based on PVC cages surrounded by a net mesh and buried in sand were designed to determine the causes of important mortalities in producing locations of carpet shell clam, Ruditapes decussatus, in two important production areas of Galicia (NW Spain). The pathogenic status of the clam populations (focusing mainly on Perkinsus sp. and clam haplosporidian) as well as changes in water temperature and salinity were recorded. Sampling was conducted during a short period of time when high mortalities were present in the two locations and in a 1‐year study when no important mortalities were registered. We found that the environmental conditions and not the presence of pathogens seemed to be related to mortalities in the study. Our results suggest that mortality rates, especially in natural beds, can emerge from a complex synergy of different factors. Moreover, we propose in situ controlled systems based on PVC cages as a useful method to control infaunal bivalve mortalities in natural beds.     

Application of enzyme-linked immunosorbent assay (ELISA) for the study of reproduction in the Manila clam Ruditapes philippinarum (Mollusca: Bivalvia): II. Impacts of Perkinsus olseni on clam reproduction

We investigated the effects of infection by the protozoan Perkinsus olseni on the reproduction of female Manila clams, Ruditapes philippinarum, from a population in Gomso Bay, Korea. The reproductive effort of the clams was assessed by ELISA using a clam egg-specific antibody and was expressed as a weight-based gonadosomatic index (GSI). The number of Perkinsus infecting each clam was estimated from the gills using Ray's fluid thioglycollate medium (RFTM) along with a NaOH digestion assay. We found that reproductive effort was negatively correlated with the intensity of the Perkinsus infection: more heavily infected clams produced fewer eggs during the spawning period from May to August. Frequency of spawning was also negatively correlated with the level of Perkinsus infection; heavily infected clams (HIC) exhibited a single spawning pulse in late July, whereas lightly infected clams (LIC) showed three spawning peaks in mid-May, late July, and late August. Egg production of HIC was only 30–75% of LIC during spawning. The level of total protein in LIC was also higher year round than in HIC. In conclusion, our investigation demonstrates that a high level of Perkinsus infection affects spawning frequency and reduces egg production, which may have long-term impacts on clam recruitment and population growth.     

Growth,mortality and reproduction of the transplanted Manila clam (Ruditapes philippinarum Adams & Reeve 1850) in Jiaozhou Bay

Samples of Manila clam (Ruditapes philippinarum Adams & Reeve 1850) were collected from May 2004 to April 2005 monthly, and plankton net trawling of planktonic larvae and bottom sediment sampling surveys were further conducted from May to October 2006 in Jiaozhou Bay. Based on the data collected, growth, mortality and reproduction of the transplanted Manila clam and the environmental effects were examined. The results showed that the enhanced clams grew well and showed a growth trend similar to the local wild ones. The main growth periods lasted from April to September, with the water temperature being the main factor affecting the growth, which was the same as that of the wild clams. There were also two reproduction cycles for the farmed Manila clams each year in Jiaozhou Bay and the main breeding period was from May to June. The phenomenon of delayed metamorphosis was quite common through larval development. The farmed clams could spawn when they reached sexual maturity, but they could not perform effective recruitment as many planktonic larvae died during metamorphosis and settlement. A preliminary study indicated that sediment perturbation and marine environment pollution were the main factors causing the death of larvae in the development process.