Étude de bactéries associées aux élevages de larves de bivalves marinsStudy of bacteria associated with the rearing of larvae of marine bivalves

Eighty-one bacterial strains associated with the rearing of marine bivalve larvae have been isolated from different origins: unicellular algal cultures; filtered sea water; unhealthy larvae; Mytilus edulis spawn, and contaminating strains from the laboratory atmosphere.The ecological, biochemical and nutritional characteristics and the sensitivity to antibiotics of these strains have been studied with the aid of 115 tests. This population appears to be similar to comparable populations studied by other authors.However, complementary studies taking into account the quantitative and qualitative variations of each source of bacterial contamination have been shown to be necessary, and are now in progress. When complete, more efficient bacterial control of the breeding of marine bivalve larvae will be possible.     

Recirculation nursery systems for bivalves

In order to increase production of bivalves in hatcheries and nurseries, the development of new technology and its integration into commercial bivalve hatcheries is important. Recirculation aquaculture systems (RASs) have several advantages: high densities of the species can be cultured resulting in a cost-effective production system; optimal temperature maximizes production and allows rapid turnover of the product; stable water quality improves growth rate and minimizes stress and potential loss by diseases. Pilot RAS systems were developed for seed rearing of oysters (Crassostrea gigas), scallops (Pecten maximus), mussels (Mytilus edulis) and clams (Ruditapes decussatus). Optimal feed addition and waste matrix were determined. Based on this, system flow rates were designed. Seed growth in the pilot RAS systems was compared at different renewal rates and with growth in flow-through systems (FTS). All four species can be reared in RAS and showed similar growth in RAS and in FTS or in RAS with a higher renewal rate. RAS can keep O₂, nitrogen and pH within the desired range. Temperature was generally higher in RAS than in FTS, probably due to heat induced by the pump circulating the water. The supply of sufficient amount of food in combination with a desire to reduce the renewal rate calls for use of concentrated feed in RAS.     

The effect of live feeds bathed with the red seaweed <Emphasis Type="Italic">Asparagopsis armata</Emphasis> on the survival,growth and physiology status of <Emphasis Type="Italic">Sparus aurata</Emphasis> larvae

Larval rearing is affected by a wide range of microorganisms that thrive in larviculture systems. Some seaweed species have metabolites capable of reducing the bacterial load. However, no studies have yet tested whether including seaweed metabolites on larval rearing systems has any effects on the larvae development. This work assessed the development of Sparus aurata larvae fed preys treated with an Asparagopsis armata product. Live prey, Brachionus spp. and Artemia sp., were immersed in a solution containing 0.5% of a commercial extract of A. armata (Ysaline 100, YSA) for 30 min, before being fed to seabream larvae (n = 4 each). In the control, the live feed was immersed in clear water. Larval parameters such as growth, survival, digestive capacity (structural-histology and functional-enzymatic activity), stress level (cortisol content), non-specific immune response (lysozyme activity), anti-bacterial activity (disc-diffusion assay) and microbiota quantification (fish larvae gut and rearing water) were monitored. Fish larvae digestive capacity, stress level and non-specific immune response were not affected by the use of YSA. The number of Vibrionaceae was significantly reduced both in water and larval gut when using YSA. Growth was enhanced for YSA treatment, but higher mortality was also observed, especially until 10 days after hatching (DAH). The mortality peak observed at 8 DAH for both treatments, but higher for YSA, indicates larval higher susceptibility at this development stage, suggesting that lower concentrations of YSA should be used until 10 DAH. The application of YSA after 10 DAH onwards promotes a safer rearing environment.     

Nutrition and feeding research in the spotted rose snapper (<Emphasis Type="Italic">Lutjanus guttatus</Emphasis>) and bullseye puffer (<Emphasis Type="Italic">Sphoeroides annulatus</Emphasis>), new species for marine aquaculture

The spotted rose snapper (Lutjanus guttatus) and bullseye puffer (Sphoeroides annulatus) are fish species from the tropical Eastern Pacific for which controlled production of larvae and juveniles has been accomplished in recent years. Diverse topics relating to their biology and aquaculture production are currently under study, in particular the nutrition and feeding aspects required to formulate practical feeds and rearing protocols. Improvements in larval growth and survival are possible by feeding live food organisms with natural or enhanced essential fatty acids content and highly digestible artificial microdiets. The ontogeny of the digestive tract and the expression and activity of digestive enzymes have been described for S. annulatus larvae. The effect of various protein and lipid levels on growth and feed utilization has been studied in juvenile and on-growing fish. Both species have carnivorous feeding habits and require high levels of protein in their diets, from 40% to 45% (dry weight) in spotted rose snapper and above 50% in bullseye puffer, with the younger stages requiring the highest protein levels. Encouraging results have been obtained in feeding experiments with different sources of dietary protein from animal and plant origin to evaluate their suitability as feed ingredients in practical diets. Optimization of fish culture practices through feeding management has also been investigated. Trials with various fish densities and feeding frequencies in intensive culture systems are providing information to improve feed utilization and growth in on-growing fish. Further research is underway to evaluate factors in broodstock nutrition which have an impact on egg and larval quality, and into the use of various commercially available oil sources in on-growing diets. In this paper, the results on nutrition and feeding research with both species are reviewed and research needs to support their commercial production in the region are discussed.     

Rearing river lamprey <Emphasis Type="Italic">Lampetra fluviatilis</Emphasis> (L.) larvae under controlled conditions as a tool for restitution of endangered populations

A method for actively protecting river lamprey and increasing the number of river lamprey populations is presented as protocol of rearing stocking material. The objective of the study was to accommodate lamprey larvae during the most critical life period under controlled conditions until they reach a sufficient size, which will increase their chances to survive in the natural environment. The eggs obtained from wild breeders were incubated in Weiss’s jars at the water temperature of 12 °C. The larvae were reared in tanks set up in closed recirculating aquaculture systems. During the rearing period, the water temperature was 20 °C and the density of larvae was 150 individuals per 1 dm² of bottom. In the initial rearing phase, the main focus was on the type of feeds and the nourishment method. It was determined that the most suitable feed type was Artemia nauplii combined with Hikari dry food. Additionally, the type of substrate was tested under controlled conditions. In this case, the optimum one was composed of sand of the grain size between 0.3 and 0.5 mm. The experiment proved that it is possible to successfully conduct river lamprey larvae rearing under controlled conditions. The growth rates are comparable to or better than those achieved under natural conditions. After 30 days of the rearing period, the lamprey larvae were 25 mm long. The results create a possibility to develop a technology for the restitution of river lamprey and other lamprey species, which are endangered throughout the world.     

Remarkably high survival rates under dim light conditions in sutchi catfish <Emphasis Type="Italic">Pangasianodon hypophthalmus</Emphasis> larvae

The sutchi catfish Pangasianodon hypophthalmus is an important species for aquaculture in Southeast Asia. However, larvae typically have low survival rates due to their predilection for cannibalism during rearing. This study investigated larval feeding behavior and rearing conditions. The former experiments were performed to elucidate the role of sensory organs in feeding, and the latter experiments examined the effect of dim light on larval rearing and survival rates. Neither lighting conditions (light and dark) nor blocking of free neuromasts by streptomycin had significant effects on feeding behavior. Therefore, the feeding behavior of sutchi catfish larvae most likely depends on chemosensing rather than visual sensing. In the later experiments, larval rearing at 0.1 l× yielded significantly higher survival rates than other lighting conditions (0.00, 1, 10, 100 l×). Survival in the 0.1 l× group was almost three times higher than in the 100 l× group. Moreover, larvae reared under 0.1 l× showed steady growth. Therefore, it is concluded that 0.1 l× is the optimum light intensity for sutchi catfish larval rearing.     

Efficiency of stagnant water larviculture using disinfected amictic eggs of the rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> in Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

There is a need to develop low-cost methods for larviculture that emphasize disease prevention. We evaluated stagnant water larviculture in flounder Paralichthys olivaceus using rotifer amictic eggs treated with glutaraldehyde. Growth and survival of the larvae were similar to in larvae reared by standard running water larviculture with daily feeding of rotifers (control). In the experimental group, the n-3 highly unsaturated fatty acid content of rotifers in the rearing water was higher than in the enriched rotifers fed to the control group. We also observed a similar pattern for the fatty acid composition of flounder larvae. The viable bacterial counts associated with rotifers in stagnant water were lower than those associated with enriched rotifers, and there were fewer bacterial genera. However, the viable bacterial count was the same or higher in the experimental rearing water compared with that in the control group. The α-Proteobacteria and Cytophaga–Flavobacterium–Bacteroidetes group dominated the bacterial community structure after larval hatching. To our knowledge, this is the first time that stagnant water larviculture, using only disinfected amictic eggs, has been successfully demonstrated on a mass production scale. This method appears to offer benefits including improved nutrition, more favorable bacterial communities, and lower cost.     

Estimation of flow in a rearing tank of marine fish larvae by simplified numerical computation—a case of two-dimensional flow

Marine fish larvae are fragile against physical stress. However, few studies have been conducted to evaluate the flow field in a rearing tank, which is assumed to provide a high degree of physical stress to marine fish larvae. The flow field in a rearing tank (volume of 1 m³) is generated by aerators, which are commonly used to provide oxygen.

This paper is a report on the estimation of stationary flow in the rearing tank of marine fish larvae. The larvae are seven band grouper larvae of Epinephelus septemfasciatus, which have a very low survival rate immediately after the hatching of eggs. The experiments of rearing of seven band grouper larvae were carried out using rearing tanks with four aeration rates (1000, 200, 50 ml/min, and no aeration). The effects of aeration on the survival and floating death of seven band grouper larvae were examined. The experiments confirmed that the mass mortality of seven band grouper larvae depends on the flow rate in the rearing tank. Aeration at 200 ml/min resulted in the highest survival and growth rates of grouper larvae.

Larvae-rearing experiments provided evidence that the flow rates of the rearing tanks are very important design aspects of rearing tanks. The estimation of flow in a rearing tank for an aerating rate of 200 ml/min was carried out by numerical calculation. The computation was simplified by a two-dimensional flow based on experimental results. The calculated flow in the rearing tank was compared with the experimental one. The calculation of the stationary flow in the rearing tank showed good qualitative and quantitative agreement with the experimental results. The numerical estimation of the flow in a rearing tank of marine fish larvae was confirmed to be effective and satisfactory for the design of a tank that would provide optimum performance.     

Mortality control of scallop larvae in the hatchery

Larval mortalities occurring in molluscan hatcheries have often been associated with bacterial contamination, and more specifically with vibrios. Although batches of oyster and clam larvae have been routinely reared in the hatchery of Argenton (North Brittany, France) without antibiotics, high larval mortalities have been recorded with the great scallop, Pecten maximus, under similar conditions. For this species, an addition of antibiotics was found necessary and chloramphenicol was used at a concentration of 8 mg l^–1. However, this chemical has now been banned in Europe, making either substitution products or an improvement in the rearing procedures essential. Studies carried out have shown that neither a decrease in larval density (to 1 larva ml^–1) nor an increase in seawater change frequency (to one per day) had any positive effects. Furthermore, elective substances such as sugars were not suitable and the use of another antibiotic, erythromycin, led to inconsistent results. The only positive effects were obtained with lower levels of chloramphenicol, which does not resolve the problem. Because no alternative solutions have as yet been found, further research needs to be undertaken.     

Effect of salinity on occurrence of notochord deformities in Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> larvae

One of the difficulties to rear Anguilla japonica larvae is the frequent occurrence of notochord deformities. We tested the effect of salinity on the occurrence of the deformities, because we have been using 50 % diluted seawater (50 % SW) for glass eel production, on the basis of the fact that intermediate salinity saves energy due to lower cost for osmoregulation and contributes higher survival and growth rates. We reared 6-day-old larvae in 50 and 100 % SW for 85 days and observed their morphology. The occurrence rate of deformed larvae, including kyphosis and scoliosis, was significantly higher in 50 % SW (35.8 %) than in 100 % SW (25.4 %), while survival rate was significantly higher in 50 % SW (69.8 %) than in 100 % SW (32.3 %) and growth in 50 % SW (mean body depth: 7.9 ± 5.3 mm) was better than in 100 % SW (6.8 ± 4.6 mm). We speculate that the most of severely deformed larvae could not survive in the tougher condition of 100 % SW, showing the lower occurrence of deformed larvae. Eventually, the yield of normal larvae after 85 days rearing was 1.9-fold higher in 50 % SW than in 100 % SW, implying that the advantage of 50 % SW for rearing eel larvae outweighs the risk of deformities.     

Horizontal and vertical variations in survival rates of juvenile bivalves,Anadara kagoshimensis (Tokunaga, 1906), Ruditapes philippinarum (Adams & Reeve, 1850) and Atrina sp., experimentally caged in the inner area of the Sea of Ariake,Japan

International Aquatic Research - In the inner area of the Sea of Ariake, rearing experiments were conducted to examine how the survival rates of juveniles of three bivalve species (Anadara...     

Protected areas and native freshwater bivalves are not in the same place in south-east Brazil

1. Bivalves are important components of freshwater ecosystems; however, they are also one of the most threatened animal groups, especially members of the order Unionida. The main threats to freshwater bivalves are habitat modification and invasive species. Protected areas are a common way to minimize impacts and preserve native species, but they are rarely designated with a focus on freshwater invertebrates.
2. The main goal of this article was to describe the distribution of freshwater bivalves in the state of Rio de Janeiro, and the relationship between these species and bioclimatic areas, land use and protected areas.
3. Five native and two invasive bivalve species have been reported in Rio de Janeiro. They occur mainly in bioclimatic areas related to the Paraíba do Sul River and Campista Lowlands. The few records of bivalves inside protected areas are in areas of sustainable use that offer a lower level of protection, and frequently in sympatry with invasive bivalves.
4. Thus, the established protected areas in Rio de Janeiro are not adequately effective for freshwater bivalve conservation, and some species remain under threat even within them. Freshwater bivalves thus remain largely unprotected in the state.
5. The development of new protected areas and management plans should consider other faunal groups that are usually ignored, such as freshwater bivalves, to achieve more inclusive and effective protection.

     

Bacterial nutrition of great scallop larvae

Bacterial nutrition of great scallop, Pecten moximus, larvae was investigated using the radioactive tracer technique. The bacterial labelling was studied initially to obtain a high and stable specific radioactivity (¹⁴C) of bacterial cells. A higher bacterial specific ¹⁴C activity was obtained when the tracer (amino acid) was introduced in the culture medium at the beginning of the exponential growth phase. After a 12 h labelling period in a rich nutritive medium, the bacteria were depurated in seawater for 5 h (chase) to prevent further ¹⁴C excretion and then added to larval rearing vessels. The larval labelling was followed for 12 h and then larvae were placed in new vessels without radioactive bacteria. The depuration of larvae was followed for 3 days. Data obtained on ingestion and assimilation efficiency show that bivalve larvae are able to ingest and digest bacteria.     

Influence of stocking density and type of feed on the rearing of crucian carp, <Emphasis Type="Italic">Carassius carassius</Emphasis> (L.), larvae under controlled conditions

Two experiments were conducted into the rearing of crucian carp larvae under controlled conditions in an experimental closed water system. In both cases, the rearing lasted 21 days. The first experiment concerned the initial stocking density (from 50 to 600 individuals per l), whilst the second one studied the first food offered (two types of Artemia nauplii, decapsulated Artemia cysts and three types of commercial feeds) which were applied at the moment of exogenous feeding commencement. The best results were obtained using 50 larvae per l, whilst there were no significant differences within the range 200–600 larvae per l. The application of dry feed had a negative influence on the survival and other parameters. The highest survival rate was found in the groups fed with freshly hatched Artemia naupli. The results obtained indicate that the rearing of crucian carp larvae may be successfully conducted in very high stocking densities. However, the very high sensitivity of crucian carp larvae to the type of food offered during the initial days of life should be taken into account. The data presented in this article could be very useful in crucian carp larviculture.     

Two monoclonal antibodies for the recognition of Mytilus spp. larvae: studies on cultured larvae and tests on plankton samples

Monoclonal antibodies (mAbs) were generated against 2-day-old mussel larvae in an attempt to develop a rapid and rigorous method for the identification of mussel larvae in field plankton samples. Previously, we have shown that two of these mAbs recognised Galician Mytilus galloprovincialis obtained from monospecific cultures, but did not recognise the larvae of other bivalve species present in that area. To assess the possibility of using these mAbs in routine assays for measuring the abundance of mussel larvae in plankton, studies on cultured mussel larvae, at different stages of development, and tests on bivalve larvae from plankton samples were carried out. Initially, to see whether the two mAbs also recognise other mussel larval stages, they were tested against mussel larvae of different ages obtained from monospecific cultures. The results indicate that both antibodies stain all the stages tested, even 1-month-old postlarvae. In addition, we also demonstrate that these mAbs also recognise other forms of Mytilus. Both antibodies bind to M. galloprovincialis larvae from the Mediterranean Sea and M. edulis larvae. Finally, and more significantly, studies on field plankton samples were performed to confirm if both mAbs are really mussel-specific, and do not cross-react with larvae of any other bivalve species existing in the plankton. The results presented here clearly indicate that our two monoclonal antibodies specifically recognise the mussel larvae in field plankton samples from different geographical regions, but not the larvae of any other bivalve species. Thus, these monoclonal antibodies could be used for routine monitoring of mussel larvae in plankton samples from different sources.     

贝类幼虫面盘纤毛脱落病的起因及防治

为研究文蛤育苗过程中幼虫病害及其主要致病菌,实验通过构建细菌16S rDNA克隆文库、病原菌分离纯化、人工感染和药敏实验等方法对育苗场发病的浮游期文蛤壳顶幼虫样品进行了系统分析。结果显示,发病的文蛤壳顶幼虫细菌群落多样性低,地中海弧菌占比高达75%以上,推断其可能为引发此次幼虫发病的主要致病菌。从发病幼虫的匀浆组织中分离获得该优势菌株,测序及系统发育鉴定为地中海弧菌。人工感染实验确定了其致病性,菌株US2-01在1.0 × 10⁶ CFU/mL 的菌液浓度下浸泡感染文蛤浮游幼虫,96 h累计死亡率为84%。药敏实验表明,地中海弧菌菌株US2-01在12种抗生素的测试中对常用的青霉素、氨苄西林、红霉素等抗生素具有一定的耐药性,对四环素和多西环素中度敏感,对头孢他啶、庆大霉素、卡那霉素等其余7种抗生素呈现高度敏感。本研究首次报道了地中海弧菌为文蛤浮游期幼虫致病的一种潜在病原,研究结果可为文蛤幼虫疾病研究及贝类苗种培育过程中的病害防控提供科学依据。     

Maintenance and culture of living benthic molluscs from high Antarctic shelf areas

Abstract. Since October 1986 living benthic molluscs from the eastern Weddell Sea, Antarctica (Fig. 1) have been kept alive in aquaria. Special equipment and methods have been used to reduce disturbance and laboratory effects during the long-time rearing. Autecological investigations were carried out on behaviour, reproduction biology and growth of several polyplacophoran, gastropod and bivalve species.     

Effects of hydrodynamic factors on Pecten maximus larval development

Hatchery production of great scallop, Pecten maximus, remains unpredictable, notably due to poor larval survival. Large‐scale flow‐through systems up to 3500 L have been developed to avoid the use of antibiotics in static systems. Alternatively, small‐scale flow‐through systems have been successfully applied for oysters but they proved to be unsuitable to rear scallop larvae. By focusing on physical factors presumed to limit P. maximus larval development, this study aimed to optimize great scallop larvae rearing parameters under controlled conditions. First, the influence of aeration on larval performances, energetic metabolism and antioxidant defences were studied both in static and flow‐through systems. Aeration depressed larval food intake, regardless of the intensities of flow tested (100 ml/min, 155 ml/min and 270 ml/min). On the other hand, antioxidant enzyme activities remained constant or decreased, suggesting that antioxidant defences were inactivated. The increase in citrate synthase activity suggested an increase in metabolic rate possibly due to a turbulent stressful environment. All larvae exposed to such turbulence died before reaching metamorphosis, whereas those reared without aeration survived well (≈ 95%). The effects of water renewal were thereafter studied in 50‐L flow‐through flat‐bottomed tanks. No differences in survival (20.4 ± 0.5%), growth (3.8 ± 0.2 μm/d), competence (5.6 ± 0.2%), energetic metabolism level and antioxidant enzyme activities were observed when comparing 12.5 and 25 L/hr water renewal. Whereas air bubbling leads to detrimental effects, flow‐through in small flat‐bottomed tanks appears to be a suitable technique for scallop larvae rearing.     

Intensive rearing system for fish larvae research: I. Marine fish larval rearing system

Larvae nutrition and in general larvae culture is considered to be the ‘bottle neck’ for marine finfish culture. Fish larvae rearing experiments investigating nutritional factors or rearing protocols are carried out in various systems, from small beakers to very large commercial tanks, making it difficult to compare data across systems.

A continuous supply of live or dry feeds and a controlled environment, i.e. temperature, filtration, photoperiod, oxygen and pH, are essential for any experimental or commercial system. These environmental factors are best controlled automatically in order to minimize variations between tanks. However, only a few automatic systems have been developed for marine finfish hatcheries.

An experimental larval rearing system was developed to reduce variability amongst tanks (due to manual feeding and other parameters) and enhance control of environmental parameters while reducing the workload. The system includes 24 conical tanks with the option of either an up-welling or bottom draining flow through water delivery system. The inlet water passes through a gas exchange column that saturates the water with dissolved oxygen and stabilizes the pH. The system was originally designed for nutritional experiments using formulated feeds. The use of an up-welling water inlet method extends the suspension time of inert particles in the water column and also helps to suspend very small or passive swimming larvae. However, when the system is used to grow-on larvae or juvenile fish it can easily be switched to bottom draining to provide self-cleaning water dynamics for high organic loads.

A unique outlet filter was developed that eases the daily routine of replacing screens when enriched live food is used. This filter can be exchanged with a screened standpipe and outlet surface skimmer when the bottom draining flow characteristics are engaged.

The system is fully controlled by a single programmable logic controller (PLC). The PLC controls the light intensity, photoperiod, dimming time, live food and algae pumping intervals, substantially reducing labor requirements.     

Larval rearing of calico scallops, Argopecten gibbus, in a flow-through system

A flow-through (FT) culture system is described for calico scallop, Argopecten gibbus, larvae. Its performance was assessed by larval survival rate, shell growth, settlement rate and post-larval shell growth for the duration of larval life (13 days). Comparisons were made with larvae reared in standard static system (S). Effect of increased larval density on FT performance was also investigated. With comparable larval densities, survival rate of Day 2 larvae to pediveliger stage was similar in both larval rearing systems. Shell growth for FT-reared larvae was comparable or significantly higher than in the static system (P < 0.01). Settlement rate of pediveligers was comparable for both systems, averaging 30.7%, and no significant difference was seen in shell growth of FT- and static-reared pediveligers. Increased initial larval density did not affect survival rate in FT, but did negatively affect larval shell growth, settlement rate and post-larval shell growth yielding lower growth and minimal settlement rate (10.9 ± 2.8%) compared to the static system. This FT system was successful as larval rearing system, optimising space allocation in the hatchery, reducing labour, and eliminating the use of antibiotics. Optimising initial larval density within the system needs to be investigated in association with food ration.