Analysis of sinking death using video images of the swimming performance of Pacific Bluefin tuna (Thunnus orientalis) larvae

In this study, we aimed to clarify the mechanism of sinking death during the larval stage of Pacific Bluefin tuna Thunnus orientalis by investigating the effects of swimming performance on sinking death, using a behavioral approach. Swimming performance was examined 3–9 days after hatching (DAH) under day and night light conditions in cuboid experimental tanks. Swimming behavior variables such as swimming speed and swimming angle were measured under both light conditions. Larvae in the daytime experiment and larvae with inflated swim bladders at night were distributed on the surface layer of the water column. In contrast, larvae with uninflated swim bladders at night were frequently observed swimming vertically or sinking to the bottom of the tank. Larvae with inflated swim bladders at night were always distributed beneath the surface until the next morning (survival rates were 100 %). However, larvae with uninflated swim bladders at night swam upward repeatedly and later sank to the bottom of the tank (survival rates were 60 % and 38 % at 5 and 9 DAH, respectively). Larvae with uninflated swim bladders were not always able to maintain their swimming depth by swimming until the next morning. Additionally, their swimming speed and vertical swimming frequency (ratio) depended on the illumination and swim bladder conditions. Our findings show that larvae with uninflated swim bladders at night were associated with a higher risk of sinking death. The swimming energy capacity of Pacific Bluefin tuna larvae, which indicates the total amount of the energy that enables individuals to swim throughout the night without feeding, was found to be linked to sinking death.     

Effect of water surface condition on survival,growth and swim bladder inflation of yellowfin tuna,Thunnus albacares (Temminck and Schlegel), larvae

Early‐stage mortality due to surface water tension‐related death and due to sinking to the tank bottom was investigated for yellowfin tuna, Thunnus albacares (YFT), larvae. Different aeration rates and rearing water surface conditions were examined to evaluate the effect on larval survival, swim bladder inflation and growth. The percentage survival of yolk sac larvae was significantly higher when the rearing water surface was covered with fish oil at aeration rates of 0 and 50 mL min^?1. The highest mortality occurred at the highest aeration rate of 250 mL min^?1 regardless of surface water condition. A second experiment was conducted twice under different water surface conditions: the water surface was covered by fish oil (FO), skimmed of fish oil (SS), and was not treated (NC). The percentage survival was not significantly different between treatments after 7 days of feeding. In contrast to the survival, the proportion of larvae with inflated swim bladders was significantly higher for the NC and SS groups than that of the FO group. Results of these experiments indicate that the addition of oil to the rearing water surface without its removal interferes with the initial swim bladder inflation in YFT larvae. These results also indicate that YFT larvae need to obtain (gulp) air at the water surface for initial swim bladder inflation, and success of initial swim bladder inflation may be crucial for their survival.     

Ontogeny of swim bladder inflation and caudal fin aspect ratio with reference to vertical distribution in Pacific bluefin tuna Thunnus orientalis larvae

It is necessary to understand the processes involved in sinking death in Pacific bluefin tuna Thunnus orientalis aquaculture in order to develop methods to prevent or minimize this problem. We observed the nighttime vertical distribution of Pacific bluefin tuna in the water column on 2–9 DAH and the morphological characteristics of the larvae, in order to clarify the processes involved in sinking death. A cuboid tank (height 300 cm) was used to measure vertical distribution. The number of larvae was counted in each of 4 regions in the observation tank: upper layer (water depth 0–100 cm), middle layer (100–200 cm), lower layer (200–300 cm), and bottom area. The distribution of larvae in these regions at 4 days after hatching was polarized to the upper layer and bottom area. Individuals with inflated swim bladders were observed in the upper layer 3 days after hatching. No larvae with inflated swim bladders were observed in the bottom area on any day after hatching. Total body length and caudal fin aspect ratio of larvae with both inflated and un-inflated swim bladders were greater in the upper layer than those of larvae in the bottom area. Larvae with un-inflated swim bladders that failed to develop sufficiently for swimming sank to the tank bottom and died. Swim bladder development and caudal fin swimming ability are strongly related to sinking death.     

Mortality,growth and swim bladder stress syndrome of sea bass (Dicentrarchus labrax) larvae under varied environmental conditions

Responses to variations in light, temperature and salinity by sea bass larvae are described here. Massive mortality of larvae has been common in Mediterranean culture efforts. This mortality, associated with swim bladder malfunction, has been analysed regarding its association with individual stresses.Optimum temperatures for sea bass growth are clearly unsuited to larval rearing, while brackish sea water (25‰) just as clearly improves growth and survival. Light and nutritional regimes as well as water quality are other sources of environmental stress that may induce the swim bladder stress syndrome (SBSS) and reduce growth and survival. SBSS should not be confused with “gas bubble” disease and its most effective treatment depends on an understanding of larval environmental requirements.Larval growth was enhanced by both decreased salinity and increased temperature, but not by increased light. SBSS and mortality were avoided only when larvae were held at ambient temperature (12.5°C), natural light, and reduced salinity (25‰). Larvae dying with distended swim bladders also had other symptoms of environmental stress—spinal abnormalities, calculi in the urinary bladders, copious mucous and opaque (edemous) tissues. Elevations of temperature and light, as well as ambient sea water (36‰) are environmental stressors and induce SBSS.     

Influence of initial swimbladder inflation failure on survival of Pacific bluefin tuna,Thunnus orientalis (Temminck and Schlegel), larvae

This study investigated the influence of initial swimbladder inflation (ISI) failure on survival of Pacific bluefin tuna (PBT), Thunnus orientalis, larvae to prevent mass mortality due to sinking death. In Experiment 1, swimbladder inflation frequency and survival within an ISI promoted (PS) group, for which surface film of rearing water was removed, and a group without ISI promotion (NPS) were compared in 20 and 30 kL tanks. The PS group demonstrated significantly higher swimbladder inflation frequency and increased survival than the NPS group within 20 kL tanks at 9 days post hatching (dph). Similar tendencies were observed within 30 kL tanks. In Experiment 2, larval distribution and swimbladder inflation frequency in the night‐time were examined through larval sampling within the upper and middle layers of the water column and tank bottom within 1.6 and 30 kL tanks at 5 dph. Larvae at tank bottom had higher distributional density and significantly lower swimbladder inflation frequency than those distributed in the upper and middle layers within 1.6 kL tanks. Similar tendencies were observed within 30 kL tanks. Results of this study suggest the improvement of larval survival due to prevention of sinking death through ISI promotion in mass‐scale PBT larviculture.     

Initial swim bladder inflation in the larvae of Tilapia mossambica (Peters) and Morone saxatilis (Walbaum)

Tilapia and striped bass larvae inflate their swim bladders on the 7th–9th and 5th–7th days after hatching, respectively. The primordial bladder of Tilapia has no pneumatic duct and larvae do not gulp atmospheric gas for the initial swim bladder inflation. Prominent columnar epithelium is characteristics of the bladder of Tilapia prior to inflation. Hypoxic conditions inhibit inflation and result in degeneration of the columnar epithelium and the irreversible malfunction of the swim bladder. The swim bladder of the striped bass has a glandular epithelium and a pneumatic duct prior to inflation. The mode of the initial inflation and functional role of the pneumatic duct remain unclear, although strong aeration and turbulent water in the rearing containers were observed to be main factors enhancing normal inflation of the larval swim bladder.     

Swim bladder inflation and survival of Mugil cephalus to 50 days

Excessive inflation of the swim bladder causes immobility and mortality of 12-day old larvae of Mugil cephalus and other marine fish. Increased aeration or mechanical agitation of the water in rearing tanks maintained the larvae below the water surface, and the swim bladder formed and functioned normally. Survival was dramatically increased. Two experimental production tanks, stocked initially with 15 000 and 20 000 larvae, at a density of 5.25 and 7.0/l, respectively, yielded 33.5% and 16.9% juveniles on day 50. Only an outbreak of ciliated parasites prevented both tanks from having survival figures of over 30%.     

Characterization of swim bladder non-inflation (SBN) in angelfish, Pterophyllum scalare (Schultz), and the effect of exposure to methylene blue

Failure to inflate the swim bladder is regarded a major obstacle in the rearing of many fish species. We present a study of swim bladder non-inflation (SBN) in angelfish, Pterophyllum scalare. A normal developing primordial swim bladder was first discernable at the end of the first day post-hatch (p.h.) as a cluster of epithelial cells with a central lumen, surrounded by presumably mesenchymal cells. Initial inflation occurred on the fourth day p.h. Prior to inflation the swim bladder epithelium consisted of an outer squamous and inner columnar layer. Cells of the inner layer were filled at their basal region with an amorphous material, which disappeared upon inflation. A pneumatic duct was absent, and larvae presented no need to reach the water surface for inflation, suggesting that angelfish are pure physoclists. A model for the role of the amorphous material in normal initial inflation is proposed. Abnormal swim bladders were apparent from the fourth day p.h., and methylene blue (MB) at a concentration of 5 ppm significantly increased the prevalence of SBN. Histologically, abnormal swim bladders in larvae hatched in 5 ppm MB could not be distinguished from those in fish raised under routine conditions (0.5 ppm MB). We suggest that MB may have a teratogenic effect in angelfish.     

Combined Effects of Diet and Salinity on European Sea Bass Larvae Dicentrarchus labrax

Effects of diet, salinity, and consumption of endogenous reserves on the growth, survival and swim bladder inflation of European sea-bass larvae were studied. Primary inflation of the swim bladder occurs in 7-day-old larvae. Inflation is synchronized with the absorption of endogenous reserves. The kinetics of the consumption of these reserves varies with the diets tested, and is the slowest at higher salinity (28.7 and 37.7‰). Best results were obtained with larvae fed on Artemia nauplii. Larvae raised at lower salinity (19.7 and 10.7‰) had the highest proportion of inflated swim bladders. Larvae raised at 28.7‰ salinity had the best survival rate, while those raised at 38.7‰ had the best growth rate. Larval mortality was associated with inappropriate functioning of the swim bladder. Larvae exposed to decreasing salinity during rearing showed very high survival rates and normal swim bladder inflation.     

Dynamics of yolk sac and oil droplet utilization and behavioural aspects of swim bladder inflation in burbot, Lota lota L., larvae during the first days of life, under laboratory conditions

The study concerns investigation of early larval development of burbot, Lota lota. As part of a two-year study, the first 15 days (until the end of yolk sac resorption) of burbot larvae development under controlled conditions were examined. The aim of the study was to observe the moment of swim bladder inflation and the behavioural aspects of this process, as well as the analysis of yolk and oil droplet resorption and the beginning of exogenous nutrition. It was observed that larvae began to inflate their swim bladder on the 3rd day post-hatch. On 5 DPH, none of the larvae without an inflated bladder was able to swim up the distance separating it (10 cm) from the water surface. Since 9 DPH, 50 % of larvae started exogenous feeding, and the absence of yolk was observed on 13 DPH and oil droplet on 14 DPH, while on 15 DPH, 100 % of feeding larvae were observed. The presented results indicate that the analyzed period is critical for burbot, and it is characterized by a high mortality rate (over 60 %). Additionally, the results suggest that, under controlled conditions, the latest moment when burbot larvae should be given exogenous food is 9–10 DPH and until the moment of the swim bladder inflation, larvae should be kept in tanks with a low water depth (up to 10 cm). The data presented in this study could have a significant influence on the efficiency of larvae rearing, both for aquaculture and for restocking purposes.     

Swimbladder inflation associated with body density change and larval survival in southern bluefin tuna Thunnus maccoyii

High mortality of southern bluefin tuna (SBT) Thunnus maccoyii larvae in captivity is a major problem hindering culture of this species. The relationships between body density of SBT larvae, swimbladder inflation and survival were investigated in this study. Swimbladder inflation and changes in volume have a direct effect on body density in larval SBT. Swimbladder inflation was first observed at 3 days post hatch, and larvae with successful swimbladder inflation were able to maintain their body density within a narrow range (?ρ = 0.0006 g cm^?3). Although swimbladder volume increased with larval growth, it could not compensate for the increase in body density and did not prevent nocturnal sinking. The increase in body density was greater for larvae that did not inflate their swimbladder. Low percentages of swimbladder inflation (27.5 ± 3.5 %) coupled with negative body buoyancy of SBT might contribute to mortality as larvae sank in the dark phase and made contact with the tank bottom. Management strategies that maintain larvae within the water column and enhance swimbladder inflation are required to improve survival of SBT larvae reared in hatcheries.     

Lobster larval transport in the southern Gulf of St. Lawrence

A bio‐physical semi‐Langrangian model was developed to follow the drift and abundance of lobster larvae from hatching to settlement as post‐larvae. The geographic domain encompasses Northumberland Strait and the areas surrounding Prince Edward Island in the southern Gulf of St. Lawrence. The model was run for ten larval seasons, 1983–87 and 1997–2001, representing two periods of egg production. The model was forced with tides, winds, heat fluxes, and marine and freshwater fluxes at open boundaries. Biological inputs were location, date, and density of larvae at hatching, development rates, a time window and a minimum bottom temperature required for successful settlement, and two scenarios of daily mortality. Net drift was west to east but stronger on the north than the south side of Prince Edward Island. The hatch was greater in the second 5‐yr period, but the spatial patterns of settlement were similar. For both mortality scenarios, the same five of the 24 larval source areas were important in providing settlers. Horizontal shear of larval distribution indicates fishing communities were dependent on the hatch from fishing grounds of multiple upstream communities. Variation in daily and annual post‐larval settlement was greatest north of Prince Edward Island. From the 24 source areas there was an eightfold range in the fraction of larvae surviving to settlement, with advection into deep water an important cause of mortality. Four to five possible fishery management areas were identified using multidimensional scaling to group sink areas having shared source areas. We hypothesize leaky discontinuity in connectivity between these groups.     

Behavioural responses to visual environment in early stage Atlantic cod Gadus morhua L. larvae

Marine fish larvae exhibit species‐specific sensitivity to light, which changes during ontogeny. Our aim was to explore how visual environment, characterized by light intensity, microalgae addition and wall colour, affects spatial distribution of Atlantic cod larvae. We applied two different small‐scale factorial designs, each of them as a series of independent short‐term trials at fixed larval ages from 5 to 35 days post hatch (DPH). Positive phototaxis increased in larvae from 5 to ≈24 DPH, followed by reduced response at later ages. Effects from the experimental factors on larval behaviour changed with age, and significant interactions between factors were present. Algae significantly affected larvae only at 5 DPH, increasing phototactic response. White wall colour induced distribution of larvae close to the tank walls at all ages. The experimental approach demonstrated clear effects from visual environment on larval distribution, and revealed both relative importance of individual factors, interactive effects and temporal changes due to larval ontogeny. Our results suggest that by fine‐tuning the visual environment in tanks, the larval distribution may be managed to improve overall rearing conditions through a more even distribution with reduced closeness to tank walls.     

鳔器官的发育对人工培育鱼苗的影响

论述了鳔器官的发育对海水鱼类人工培育苗种的影响，仔鱼期鳔的形成和充气是影响闭鳔海水鱼类人工育苗成效的重要因素之一，鳔发育异常一是导致仔稚鱼死亡，二是导致脊柱前凸。在人工养殖条件下，影响海洋闭鳔鱼类鳔器官充气的因子有营养因子和环境因子，充气量，水表面的油膜，温度，盐度和光照等环境因子影响仔鱼水表面吞饮空气，影响鳔的充气率，仔鱼摄食EPA和DHA含量低的饵料后，活力差，仔鱼到达水表面吞饮空气的能力下降，鳔的开腔率降低，此外仔鱼体内甲状腺素的含量可能对仔鱼鳔的开腔率产生影响。     

Promotion of initial swimbladder inflation in Pacific bluefin tuna,Thunnus orientalis (Temminck and Schlegel), larvae

Initial swimbladder inflation (ISI) of Pacific bluefin tuna (PBT), Thunnus orientalis, larvae was studied to increase the survival of cultured larvae. Experiment 1 was conducted to explore promotion and inhibition of ISI under different water surface conditions; including the use of surface skimmer to remove autogenous surface substances (SS), covering the water surface with liquid‐paraffin‐layer (LP) and oil film (OF), and a control (non‐treatment, NT). Significantly higher inflation frequency was observed in SS (62.2%) than NT (11.9%), LP (2.7%) and OF (3.9%). This indicates that ISI in PBT larvae can be promoted by removal of surface substances on rearing water which inhibit larval air gulping. Experiment 2 aimed to elucidate proper day of larval age to start skimming for promoting ISI with four different periods of oil film removal: from 3 to 8 (SF3D), 4 to 8 (SF4D), 5 to 8 (SF5D), 6 to 8 (SF6D) days‐post‐hatch (dph). Significant improvement in ISI frequency was observed in SF3D (80.2%) but the frequency was very poor in SF4D, SF5D, and SF6D (17.8–7.5%). This implies the need of oil film removal without missing a narrow window, 1 day of 3 dph, to promote ISI in practical PBT larviculture.     

The effect of light intensity on performance of larval pike-perch (Sander lucioperca)

In this work performance parameters of larval pike-perch (Sander lucioperca) reared under four different light intensities (100, 500, 1000 and 2500 lx) until 21 days post hatch (dph) were investigated. As performance parameters change in length and weight, swim bladder inflation, feed consumption, natural mortality, stress induced mortality and RNA-DNA ratio were measured. Aim was to investigate the influence of light intensity on pike-perch performance during the first three weeks of larval rearing. Significant differences were found in natural and stress induced mortality as well as in weight growth. No single light exposure level combined optimal performance of all tested performance parameters. Highest light intensity of 2500 lx showed good weight growth but an increase in stress induced mortality. Bright light of 500 and 1000 lx intensity was found to improve growth and stress mortality whereas dim light conditions of 100 lx showed significantly lower natural mortality. Thus data suggested that most favorable illumination during larval pike-perch rearing comprise a tradeoff between optimal natural mortality under dim light conditions (100 lx) or optimal larval growth and stress resistance under bright light conditions (500 and 1000 lx). It is shown that high light intensities during larval rearing can be beneficial for pike-perch rearing if offspring supply is not limited.     

Does larval mortality influence population dynamics? An analysis of North Sea herring (Clupea harengus) time series

The daily mortality rates of North Sea herring early‐stage larvae are found to vary over decades. Larval abundance data were used with a spatio‐temporal oceanographic model to reconstruct temperature histories of the observed larvae. The histories were used in conjunction with a temperature‐based growth model to estimate larval age. Mean daily mortality rates were then estimated for the four spawning components (Downs, Banks, Buchan and Orkney/Shetland) using the vertical life table approach, which considers instantaneous abundances across all ages rather than following distinct cohorts. All spawning components, but especially Downs (in the south), exhibited a steady rise in mortality associated with increasing population size. In addition, the three northern components shared a distinct trend in mortality that was significantly correlated with ambient water temperatures experienced by the larvae during the respective time periods after hatching. This trend was also significantly negatively correlated with the residuals of the whole stock‐recruitment relationship. These findings were generally robust to assumptions about growth and hatch length of larvae. The compensatory increase in productivity in the late 1980s and poor recruitment since 2000 coincide with changes in the mortality of larvae younger than 30 days post hatch and covary with larval density and temperature. Thus we suggest that the mortality of early‐stage larvae does impact on the population dynamics in North Sea herring in its current productivity regime, implying a critical period in the determination of year class strength.     

EPIZOOTIOLOGIC STUDY OF MID-CYCLE DISEASE OF LARVAL Macrobrachium rosenbergii

Mid-cycle disease (MCD) of larval Macrobrachium rosenbergii has reduced survival of larval cultures at the state facility from a normal 50–70% of the population at the post-larval stage, to 5–10%. Observations revealed a characteristic mortality pattern, where a peak in daily mortality occurred near the middle of the rearing cycle. Epizootiologic studies were initiated using existing facilities, to aid in characterization of MCD and elucidate possible causes. Experiments were undertaken in duplicate culture tanks, each separated into 4 equal volume compartments by nitex screen. Water, algae, and some feed could pass between compartments; prawn larvae could not. “Reference” larvae were introduced into a compartment of each tank on the day of hatch. “Test” larvae populations of different ages and sources were introduced into the remaining compartments at various times after the initiation of the experiment. Dead larvae in the bottom sediment were counted daily and mortality curves generated. Length and developmental stage of live and dead animals in each compartment were assessed daily. Results indicated that a “toxic period” of unknown etiology began on the 10th day of culture and probably extended for the duration of the rearing cycle. “Test” populations introduced during the “toxic period” displayed a mortality peak within 4–6 days. Larvae at age 15 days (stage 6–7) or older appeared refractory to the “toxicity.” MCD apparently reduced growth rates (length) but did not appear to selectively kill larger, faster growing, susceptible individuals. Problematically large portions of animals lost from populations were not accounted for. Studies on MCD will continue.     

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.     

Relationship between oxygen consumption,growth and survival of larval fish

This work aimed to evaluate the relationship between morphological development, oxygen consumption and reduced mortality in larval fish. We measured the resting metabolic rate (RM), specific metabolic rate (SMR) and the change in the total length during the larval stage of four fish species. Resting metabolism decreased from hatching to mouth opening, and then increased after mouth opening. The changes in the SMR were variable during the larval stage. After hatching, there was no increase in SMR in yolk‐sac larvae. However, SMR increased between mouth opening and the onset of notochord flexion and then decreased during notochord flexion before finally stabilizing. We observed two peaks in mortality during the larval period of all species: between mouth opening to the onset of notochord flexion and from the completion of notochord flexion to the juvenile stage based on the per cent mortality and the number of dead fish collected from the bottom of the rearing tank. Interestingly, the changes in SMR coincided with these periods of mortality. We hypothesize that larvae require more energy during these periods of larval development and are thus more susceptible to mortality when energy is insufficient. Thus, it is important to supply enough nutrition to larvae in during early development to prevent mass larval mortality.