Analysis of bacterial communities in <Emphasis Type="Italic">Nannochloropsis</Emphasis> sp. cultures used for larval fish production

ABSTRACT:   Phytoplankton used in fish hatcheries is mass-cultured in the open air and usually contains large numbers of bacteria. In commercial fish production, the phytoplankton cultures are usually added into the larval rearing tanks; however, the numbers and types of bacteria introduced into the rearing tanks simultaneously are unknown. In this study, the bacterial community structures in Nannochloropsis sp. cultures were analyzed by using fluorescence in situ hybridization (FISH). A direct viable count (DVC)-FISH analysis was also performed as DVC is useful for the detection of actively growing cells. Total numbers of bacteria in Nannochloropsis sp. cultures ranged from 7.72 × 10⁵−2.39 × 10⁶ cells/mL. High proportions of the total bacteria (31.6–53.6%) in the Nannochloropsis sp. cultures showed growth potential. DVC-FISH analysis revealed that α-proteobacteria and the Cytophaga – Flavobacterium cluster were abundant in the bacterial community of actively growing cells. Thus, the high growth potentials of the distinct bacterial communities in Nannochloropsis sp. culture must influence the bacterial communities in larval rearing tanks.     

Effect of stocking density and feeding regime on larval growth,survival, and larval development of Japanese flounder,Paralichthys olivaceus,using live feeds

This research examined the effect of initial stocking density and feeding regime on larval growth and survival of Japanese flounder, Paralichthys olivaceus. Larval rearing trials were conducted in nine 50‐L tanks with different initial stocking densities combined with different feed rations (20 larvae/L with standard feed ration [LD], 80 larvae/L with standard feed ration [HD], and 80 larvae/L with four times the standard feed ration [HD+]). Larvae were stocked on 0 days posthatch (DPH) following hatching of the fertilized embryos. Larval total length (TL), survival rates, and final densities were observed on larval settlement (32 DPH) to evaluate larval rearing performance. At 32 DPH, there were no significant differences (p > .05) in TL or survival rates between the LD (46.5 ± 17.0%) and HD+ (40.3 ± 9.4%). The TL and survival rate of HD (23.1 ± 3.5%) were significantly lower than that of LD and HD+ (p < .05). However, the larval density of HD was significantly higher than that of LD (p < .05). HD+ achieved the best larvae production (32.27 ± 7.51 larvae/L), supported by sufficient food source, high water exchange, and proper water quality management (routine siphoning, surface skimming). The larval‐rearing protocols and larval development from hatching to metamorphosis is described in detail, with corresponding photographs taken during the experiment.     

Yields of great scallop, Pecten maximus, larvae in a commercial flow-through rearing system in Norway

Survival, growth and yield of competent great scallop (Pecten maximus) larvae were investigated during a full production season in a commercial hatchery in western Norway. Broodstock were collected from natural scallop beds and 12 groups were induced to spawn during the period December 2002 to July 2003. Larvae were reared on a large scale in 36 flow-through tanks (3500 l) at 17±1 °C and continuously fed a mixture of five algal species produced in an indoor continuous-flow system. Large variations in larval performance between spawning groups and tanks were observed, but the results were as good as earlier results using the batch system and prophylactic addition of chloramphenicol. Growth from days 3–24 averaged 4.8 μm day⁻¹±0.8 (sd) and survival 22.4%±21.8 (sd). Mean yield of day 3 larvae was 7.1%±10.0 (sd) and 26.6%±25.9 (sd) for those surviving to day 24. Yield was significantly correlated to larval survival. Larval success was related to initial larval density, algal concentration and season. It was found that the best production regime had an initial larval density lower than 6 ml⁻¹ and algal concentration of less than 12 μl⁻¹ regardless of season. Seventeen tanks met these criteria and produced a mean yield of 0.5 larvae ml⁻¹ to settlement. Flow-through systems are currently regarded as the only feasible method for viable hatchery production of P. maximus larvae in Norway.     

The effects of dietary lipid sources and lecithin on the production of giant freshwater prawn <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis> larvae in the Mekong Delta region of Vietnam

The giant freshwater prawn, Macrobrachium rosenbergii, is cultured widely in the Mekong Delta region of Vietnam but it is often difficult or expensive for hatchery operators to purchase commercial diets used as a feeding supplement to Artemia nauplii. Therefore, in the present study, the effects of lipid sources and lecithin on the growth and survival rate of M. rosenbergii larvae were examined in order to develop suitable hand-prepared larval diets for seed production of M. rosenbergii in this area. Six egg custard diets consisting of various ratios of lipid (originating from soybean oil and squid oil) and lecithin were used for rearing Macrobrachium rosenbergii larvae. Treatments in which larvae were fed diets containing squid oil exhibited the highest body length and survival rates (7.14–7.43 mm and 51.1–68.1%, respectively), and differed significantly from other treatments (P<0.05). Use of dietary soybean oil yielded the lowest body length and survival rates (6.29–6.75 mm and 22.0–48.7%), respectively). The supplementation of dietary lecithin did not increase final body weight but did improve larval survival rates. The n-3 HUFA content of prawns fed dietary squid oil was higher than those of animals provided with other diets. These results indicated that the most appropriate diet for rearing M. rosenbergii larvae is the diet containing 3% squid oil and 1.5% lecithin.     

Larval Age at Stocking,Growth, and Survival During Fingerling Production of the Endangered Sun Catfish,Horabagrus brachysoma

An experiment was conducted to ascertain the initial rearing period required for Horabagrus brachysoma larvae before releasing them into the nursery tank for fingerling production. Hatchlings were reared for 7, 14, or 21 days in larval rearing tanks before stocking to nursery tanks. The 7-day-old larvae were significantly smaller at the end of the 2-week nursery rearing, but over the course of nursing, the 7-day-old larvae caught up to the 14- and 21-day larvae. The specific growth rate (SGR) of 7-day larvae was highest among the three treatments. However, mortality of 7-day larvae was the highest among the groups. Hence, it appears important to rear the larvae at least 2 weeks before stocking for fingerling production so as to increase survival rate.     

Rearing performance of Clarias batrachus larvae: Effect of age at stocking on growth and survival during fingerling production

An experiment was conducted to evaluate the rearing performance of different larval stages for optimum growth and survival during fingerling production of Clarias batrachus with an aim to reduce the rearing time of the catfish larvae in a hatchery system. The larvae were reared for 5, 10 and 15 days before stocking them in the nursery tanks. No significant (P < 0.05) difference could be found in length increment at the end of first week of rearing. But the total length was reduced during rest three weeks in five days old larvae compared to that of other two higher age groups. In 5 days old larvae the reduced weight (P < 0.05) was observed during the entire rearing period. However, the specific growth rate (SGR) did not vary among the groups. The survival rate as well as total biomass was decreased while stocking five days old larvae for rearing, compared to that of other two age groups.     

Ceramic clay reduces the load of organic matter and bacteria in marine fish larval culture tanks

Ceramic clay has been increasingly used to improve contrast and prey detection in tanks for rearing of fish larvae. In contrast to live microalgae or algae pastes, clay increases turbidity without contributing to the organic matter load. In addition, clay may aggregate and sediment organic matter and bacteria, facilitating its removal. Marine larvae are sensitive to infections by opportunistic bacteria. Fish, algae, and live feed increase the microbial carrying capacity of the rearing water which allow exponential growth of bacteria and favor fast-growing opportunists. Reducing substrate levels by replacing microalgae with clay may reduce bacteria proliferation and benefit larvae. We compared the effects of three rearing regimes including live Isochrysis galbana, Nannochloropsis oculata paste, and ceramic clay on the bacterial community, concentration of organic matter, and growth and survival of Atlantic cod larvae (Gadus morhua L.). The application of clay resulted in reduced substrate levels for bacteria in the rearing water compared to the addition of live algae or algae paste. To some extent, clay aggregated and transported organic matter to the bottom of the larval fish tanks, where it could be effectively removed. Fish tanks receiving clay showed a lower abundance of bacteria in the water than tanks added algae paste or live algae. Fish tanks with algae paste showed a higher abundance of bacteria and a higher share of cultivable bacteria and TCBS counts than the other two treatments. Tanks with live algae showed low relative abundances of opportunistic bacteria and TCBS counts in both water and rotifers. Cod larvae in tanks with clay or live algae initiated exponential growth earlier than larvae in tanks with algae paste. Larvae in tanks receiving clay had significantly higher dry weight than larvae in tanks receiving algae paste at day 5 and 20 post hatching. The survival of larvae in the tanks added clay was variable. Two of the three tanks with clay had significantly higher larval survival than the tanks with live algae or algae paste. However, one tank with clay underwent 100% mortality. It is not possible to conclude whether this was related to the use of clay or an incidental development of a harmful microbial community in this tank. The effects of clay addition on larval performance should be studied further. Clay addition appears to be an easy way to reduce bacterial load during early first feeding of marine larvae without compromising the beneficial effects of turbidity.     

Development of hatchery technology for the silver pomfret Pampus argenteus (Euphrasen): effect of microalgal species on larval survival

The silver pomfret Pampus argenteus (Euphrasen) is a new candidate for aquaculture and there is not much information available on its larval rearing. Investigations carried out using microalgae alone in the culture system for the initial feeding of the silver pomfret larvae showed that Chlorella, Isochrysis and Nannochloropsis without rotifers are not conducive to the survival of newly hatched larvae. At 6 days after hatching (DAH), a maximum survival of 3% (1.8 ± 1.69%) was observed with Isochrysis followed by Nannochloropsis (0.35 ± 0.21%) and Chlorella (0.25 ± 0.21%). All control larvae died at 6 DAH without microalgae. Further investigations using the above microalgae with rotifers and a mixture of these same microalgae with rotifers showed that significantly higher (P < 0.05) survival could be achieved in the mixture of microalgae with rotifers in the culture system. At 12 DAH, the larval survival was 9.73 ± 1.39% in mixed species of algae compared with that of Isochrysis (6.93 ± 1.86%), Nannochloropsis (6.83 ± 0.61%), Chlorella (5.93 ± 2.76%) and seawater without microalgae or the control (0.73 ± 0.31%). The first incidence of feeding on rotifers at 4 DAH was significantly higher (P < 0.05) in all treatments with microalgae than that of the control. The incidence of feeding in mixed species of algae at 4 DAH (60.0 ± 0.00%) and in Isochrysis (55.0 ± 35.36%) was significantly higher (P < 0.05) than that of Chlorella (40.0 ± 0.00%) and the control (25.0 ± 7.07%). Prey consumption of individual larvae increased significantly (P < 0.01) at 8 DAH compared with that at 4 DAH. During this period, predation on rotifers by larvae was significantly higher (P < 0.05) in mixed species of algae (12.85 ± 5.73 rotifers larva^?1) than that of the control (6.75 ± 1.20 rotifers larva^?1). The fatty acid composition of rotifers used during this investigation shows that significantly higher (P < 0.05) ω3 HUFA was present in rotifers treated with mixed algae plus commercial enrichment media ‘Super Selco’ and ‘DHA Protein Selco’. Rearing of silver pomfret larvae up to the juvenile stage using mixed species of microalgae in the hatchery has been discussed. During 38 days of the larval rearing period, it was possible to achieve 3.6–4.2% larval survival with a mean of 3.9 ± 0.42%, which was considerably higher than in previous attempts (survival up to 1.5%).     

Larval survival,development and growth in the horsehair crab,Erimacrus isenbeckii,cultured under different photoperiod conditions

Larval sinking causes larval mass mortality during seed production in the horsehair crab, Erimacrus isenbeckii. Under normal light conditions, horsehair crab larvae generally show negative phototactic behaviour and sink in their rearing seawater. It has been proposed that culturing larvae in the dark may prevent larval sinking. Herein, we examined the effect of photoperiods on horsehair crab larval survival and development to facilitate the development of larval rearing techniques that prevent sinking. Newly hatched larvae were reared with Artemia to the first crab stage in 2‐L beakers under five photoperiods: 0L:24D, 6L:18D, 12L:12D, 18L:6D and 24L:0D. Larvae survived and molted to the first crab stage under all tested photoperiod conditions. The survival rate improved with increasing light period, whereas the developmental period for each larval stage decreased with increasing light period. Longer light periods increased the carapace length at the first crab stage. Our results suggest that larvae could be cultured to the first crab stage in large‐scale tanks under constant darkness. However, significantly improved larval performance under longer photoperiodic conditions indicates a need for developing alternative culture techniques to control larval behaviour in the seed production tank.     

Application of prospective probionts at early stages of Atlantic cod (Gadus morhua L.) rearing

This work aimed at validating the use of two prospective probionts (Arthrobacter sp. and Enterococcus sp.) at early stages of cod (Gadus morhua L.) rearing. Ova at late post‐fertilized stage and larvae during their first 4 weeks of life were bathed with both probionts, isolated previously from the cod‐rearing environment. This treatment was compared with groups fed rotifers supplemented with a commercial probiotic (Remus^®) and those untreated. Microbiological analyses (total viable counts, presumptive Vibrio and lactic acid bacteria) were performed in rearing systems and larval survival, growth and development were assessed. Larval development was evaluated by proteolytic activity of larval lysates and immunological analysis of important proteins: apolipoprotein A‐I, haemoglobin, C‐reactive protein, C3 and cod serum proteins. Bacterial bathing led to a significantly higher larval weight, length and culturable microbial load in larval gastrointestinal (GI) tract when compared with the control and Remus groups. Development occurred earlier in bathed larvae. However, their survival was negatively affected compared with the control group, but was significantly higher than for the Remus group. The non‐pathogenicity of both probionts was demonstrated by intraperitoneal injection of 13 g cod juveniles. The results suggest that Arthrobacter and Enterococcus probionts affected the larval GI microbiota and contributed to growth, development and digestion, either directly or indirectly.