Altered behaviour and reduced survival of juvenile olive flounder, Paralichthys olivaceus, infected by an invasive monogenean, Neoheterobothrium hirame

Neoheterobothriumhirame is a blood feeding monogenean of olive flounder Paralichthys olivaceus. The parasite was first reported in the mid-1990s from the Sea of Japan and became epidemic within cultured and wild flounder populations after several years. Infected fish often suffer from severe anaemia and thus the parasite is thought to have played an important role in the recent depletion of flounder populations in some areas of Japan. However, the causal mechanism underlying the parasite epidemic and decreases in host populations is unclear because apparently N. hirame infection is not fatal to the host. Here, we tested the hypothesis that N. hirame indirectly reduces the survival of wild juvenile flounder by altering their behaviour and making them more susceptible to predation. We conducted a series of experiments to compare behaviours and predation susceptibility between experimentally infected juvenile P. olivaceus and uninfected fish. Results showed that N. hirame infection increases the activity level, alters diel activity and has negative effects on burrowing performance and swimming endurance. When juvenile flounder cohabitated with predators, the survival rate of infected juveniles was approximately 25% less than that of uninfected fish. We believe this is the first empirical evidence linking N. hirame infection to death of the host through predation. Consequences of N. hirame-induced behavioural change for the survival of juvenile flounder in the wild are discussed. We conclude that recent outbreaks of N. hirame are likely to have been a key factor in the decline of flounder populations in Japan.     

Use of parasite and genetic markers in delineating populations of winter flounder from the central and south‐west Scotian Shelf and north‐east Gulf of Maine

Discriminant function analyses of infection parameters of parasitic helminths revealed that abundances of seven helminth species contributed significantly to the delineation of four host populations of winter flounder Pleuronectes americanus from the central and south‐west Scotian Shelf and the north‐east Gulf of Maine (NAFO subdivision 4WX‐5Z). These were adult digeneans, Derogenes varicus , Genolinea laticauda , Steganoderma formosum and Steringophorus furciger , metacercariae of the digenean, Stephanostomum baccatum , and larval nematodes, Anisakis simplex and Hysterothylacium aduncum . The correct classification rate was 84% overall, with Georges Bank (5Z) and Sable Island Bank (4W) winter flounder being the most accurately classified samples at 98 and 88%, respectively. Winter flounder from south‐west Nova Scotia (4X), an inshore sample from St Marys Bay and offshore fish from Browns Bank, had the lowest rates of correct classification (76 and 71%, respectively) due, primarily, to cross‐misclassification between the two samples. Winter pairwise comparisons of four microsatellite markers identified significant genetic differences between all populations sampled with the Georges Bank population being the most genetically distinct overall, and St Marys Bay and Browns Bank fish being the least dissimilar.     

Predator avoidance behaviour in wild and hatchery‐reared brown trout: the role of experience and domestication

Juvenile brown trout Salmo trutta from natural populations reacted to the presence of piscivorous brown trout by increasing the use of refuges. In contrast, second‐generation hatchery fish and the offspring of wild fish raised under hatchery conditions were insensitive to predation risk. The diel pattern of activity also differed between wild and hatchery brown trout. Second‐generation hatchery fish were predominantly active during daytime regardless of risk levels. Wild fish, however, showed a shift towards nocturnal activity in the presence of predators. These findings emphasize the potential role of domestication in weakening behavioural defences. They support the idea that the behavioural divergence between wild and domesticated individuals can arise from a process of direct or indirect selection on reduced responsiveness to predation risk, or as a lack of previous experience with predators.     

The cryptic responses of hatchery-reared sole to a natural sand substratum

In laboratory experiments, both reared and wild sole Solea solea selected a sand substratum in preference to a hard substratum. Reared sole with no previous experience of sand buried quickly when placed on sand. Light motivated burial, and the motivation of reared sole to bury was as strong as that of wild sole. The burial efficiency (the proportion of the ocular side covered by sand after a single burial attempt) of reared sole was lower than that of wild sole, but increased to that of wild sole after a period of 12 days maintenance on sand. Motivation to bury is therefore innate, but efficiency is affected by experience. The reactive distance of reared sole to a standardized predation threat was the same as that of wild sole and was shorter when buried (6 cm) than when not buried (15 cm). Burial therefore affected the response to a predation threat, indicating that burial is a cryptic behaviour. Munsell colour charts were used to determine the time required for reared sole to match the skin colour 'tone' of wild sole after placement on sand. Adaptation of colour value (lightness) took 4–7 days, but adaptation of chroma (intensity of colour) and hue took 33 and 69 days respectively. It is therefore recommended that flatfishes reared for stock enhancement exercises are conditioned to sand prior to release due to the relatively long time required for crypsis to improve through colour adaptation and burying.     

Field evaluation by RNA/DNA ratios on post-release nutritional status of released and wild Japanese flounder Paralichthys olivaceus juveniles

A large-scale juvenile Japanese flounder (Paralichthys olivaceus) release-recapture experiment was undertaken to find the optimal release season by evaluating the nutritional status of released fish at different seasons during which food abundance was significantly different. Forty thousand fish were released at depths of 1.5 m for early-release (May 29, 1997) and 2 m for late-release (July 2, 1997) (42.1±3.5 and 40.9±4.2 mm body length, respectively) in an experimental field, Wakasa Bay, the Sea of Japan. Samples were taken, after the releases, at Wada beach at intervals of 1, 2, 3, 6, 10, 16 and 30 days after release (DAR), including pre-surveys before each release. Released fish recaptured from the two different release groups totaled 764; 467 from the early-release group (ER) and 297 from the late-release group (LR). A total of 1956 wild flounder juveniles were simultaneously collected (1041 ER, 915 LR). ER fish were subject to higher food availability and were exposed to less pressure from predation by smaller wild juvenile flounder. RNA/DNA ratios in ER juveniles were significantly higher than those of LR fish during all samples. Especially, RNA/DNA ratios in ER juveniles were higher than in wild juveniles from 3 to 50 DAR. In the LR group, the nutritional status of juveniles was relatively low in shallower water. These findings corresponded well with feeding incidence examined by coworkers. Mass release of hatchery-reared juveniles apparently reduced RNA/DNA ratio of the wild juveniles right after releasing. The present study showed that earlier release of hatchery-reared juvenile Japanese flounder with higher RNA/DNA ratio could increase the possibilities of survival right after release in the nursery ground, and that RNA/DNA ratio appeared to be a good tool in evaluating nutritional status of released juveniles as well as wild juveniles in Japanese flounder.     

Shallow water predation risk for a juvenile flatfish (winter flounder; Pseudopleuronectes americanus, Walbaum) in a northwest Atlantic estuary

Many small fish, including several juvenile Atlantic flatfish, are most abundant in shallow areas presumable because these habitats enhance survivorship and/or growth. In this study, we investigated size-dependent depth distributions and the role of shallow habitats as predator refuges for age-0 winter flounder (Pseudopleuronectes americanus) in a northwest Atlantic estuarine nursery. Analysis of trawl surveys performed during the larval settlement period throughout the Navesink River and Sandy Hook Bay, New Jersey, showed that as fish increased in size, depth of occurrence gradually decreased, so that individuals >35 mm standard length (SL) were concentrated in habitats ∼1 m deep. Tethering in structurally simple and adjacent shallow and deep habitats showed that predation risk for flounder (30-50 mm SL) was low in shallow water (<1 m) and increased rapidly with depth. Summer flounder (Paralychthys dentatus), which were more abundant in trammel nets in deep habitats and included winter flounder in their diets, appeared to be important consumers of tethered fish. Our results indicate that following larval settlement, winter flounder emigrate from or suffer high mortality in deeper water to become concentrated in shallow habitats that can serve as predator refuges even when they lack complex physical structures. These results highlight the potential for functional habitat loss when natural and/or anthropogenic factors make shallow habitats unavailable to young fish.     

Puberty and recrudescence in cultured male yellowtail flounder: plasma androgens and spermatogenesis

Two year classes of cultured male yellowtail flounder Limanda ferruginea were followed in order to examine puberty and recrudescence at the gonadal level, and to assess the impacts of culture conditions on male reproduction. Cultured males demonstrated a propensity for maturation as 1 year‐old fish. Pubertal activation of testicular steroid production was associated with the appearance of primary spermatocytes, but a single case of a premeiotic endocrine activation was detected. The onset of puberty occurred during September and October, when adults were post‐spawned or in early recrudescence. Once initiated, pubertal development proceeded in an uninterrupted progression to full maturity by June at 22 months of age. Spermatogenetic activity during the autumn led to a peak in the gonado‐somatic index and total testicular mass by December. Milt could be first expressed from some pubertal males at this time, well in advance of the May to August spawning period in captivity. Meiotic activity could be absent from the testes by the spring, but residual areas of meiotic activity were found in pubertal males of one year class which may indicate a slower spermatogenetic rate. Androgen levels peaked during prespawning and early spawning; 11‐ketotestosterone (11‐KT) was the dominant androgen throughout spermatogenesis. Recrudescing 2 year‐old males showed greater testicular development and androgen production. Spermatogenetic patterns were similar to those seen in pubertal males, with the distinction that recrudescing males had a more discernable mitotic phase prior to meiotic activity; mitotic and meiotic activity overlapped considerably during early puberty. Testicular development well in advance of spawning may confer an evolutionary advantage to males of this cold‐water marine flatfish.     

The genetic stock structure of larval and juvenile winter flounder larvae in Connecticut waters of eastern Long Island Sound and estimations of larval entrainment

Variability at six microsatellite loci was examined among 536 winter flounder Pseudopleuronectes americanus larvae collected from three locations in eastern Long Island Sound shown to be nursery areas for the species. Substantial genetic differences were seen among the putative source populations and thus appeared to be geographically based. These differences were used to characterize the most likely sources of winter flounder larvae entrained at the Millstone Power Station as well as recruitment to juvenile winter flounder collected in the Niantic River. Samples were classified to the most likely geographical source population both by a conditional maximum likelihood method and by a multi‐layer feed‐forward neural net trained on the differences in microsatellite allele frequencies. The classification of samples by both methods is compared and discussed in the context of winter flounder management.     

Physiological and behavioural differences of hatchery and wild‐reared steelhead Oncorhynchus mykiss smolts of the same genetic origin

In many parts of the world release of hatchery‐reared smolts has long been used to mitigate for the deleterious effects of habitat loss and overfishing on salmonid populations. Of increasing concern is whether this may cause harm by spreading non‐native stocks and potentially releasing incompetent smolts. The objective of this study was to determine if smolt physiology and behavior of juveniles produced from a recently founded native broodstock differ from their wild (naturally‐reared) counterparts. In the fall of 2002 and 2003 juvenile wild steelhead were captured, PIT tagged, and returned ( n  = 1360 in 2002 and n  = 2708 in 2003) to Abernathy Creek. In winter of 2003 and 2004 hatchery‐reared fish were PIT tagged and later released ( n  = 1100 in 2003 and n  = 1400 in 2004) into Abernathy Creek. Gill biopsies were collected from wild and hatchery fish throughout the rearing and out‐migration season. The timing and speed of outmigration was assessed using two stationary PIT tag antennas (92–97% efficient). Hatchery migrants in 2003 were larger, had significantly lower gill Na⁺, K⁺‐ATPase activities, and migrated slower than wild fish. Results from the 2004 migratory season will also be presented. This study shows that hatchery rearing can result in smolts which are physiologically and behaviourally different from genetically similar wild fish. Whether these differences are critical enough to affect the rate of adult returns will be determined in future years.     

A New PCR-Based Method Shows That Blue Crabs (Callinectes sapidus (Rathbun)) Consume Winter Flounder (Pseudopleuronectes americanus (Walbaum))

Winter flounder (Pseudopleuronectes americanus) once supported robust commercial and recreational fisheries in the New York (USA) region, but since the 1990s populations have been in decline. Available data show that settlement of young-of-the-year winter flounder has not declined as sharply as adult abundance, suggesting that juveniles are experiencing higher mortality following settlement. The recent increase of blue crab (Callinectes sapidus) abundance in the New York region raises the possibility that new sources of predation may be contributing to juvenile winter flounder mortality. To investigate this possibility we developed and validated a method to specifically detect winter flounder mitochondrial control region DNA sequences in the gut contents of blue crabs. A survey of 55 crabs collected from Shinnecock Bay (along the south shore of Long Island, New York) in July, August, and September of 2011 showed that 12 of 42 blue crabs (28.6%) from which PCR-amplifiable DNA was recovered had consumed winter flounder in the wild, empirically supporting the trophic link between these species that has been widely speculated to exist. This technique overcomes difficulties with visual identification of the often unrecognizable gut contents of decapod crustaceans, and modifications of this approach offer valuable tools to more broadly address their feeding habits on a wide variety of species.     

Digestive capacity, growth and social stress in newly-metamorphosed Japanese flounder (Paralichthys olivaceus)

Hierarchies readily appear when rearing flounders through metamorphosis in space-limited conditions. In this experiment, subordinate fish were stressed, as suggested by their elevated cortisol level compared to dominant fish. Subordinate fish, although of smaller size than the dominant fish, showed no suppressed digestive capacity. By separating the two hierarchical groups into different tanks at low density, the cortisol level of the subordinate fish substantially decreased. The removal of the stressor also resulted in an increase in digestive function and improved coloration. Also, compensatory growth in length but not in weight was observed in the subordinate group, suggesting that the subordinate fish devoted their energy to increase their length. These results imply that the climax of flounder metamorphosis (i.e. the settlement stage) is a highly sensitive period, where social interactions may induce high levels of stress. However, Japanese flounder early juveniles prove to have a high recovery capacity from stress and show to allocate energy preferentially to growth in length. This seems to be an adaptation to diminish the probability of death by predation.     

Disruption of the melatonin rhythm in wild and farmed fish: a consequence of prolonged thyroxine administration and calcium depletion

Recent studies suggest that the pineal gland and its major product melatonin (N‐acetyl‐5‐methoxytryptamine: Mel) play a role in integration of various neural and endocrine functions. Observations indicate that Mel as a signal of photoperiod regulates a number of biological phenomena, including reproduction, day/night activity and many other physiological events associated with daily or seasonal rhythms in vertebrates, including fish. Mel has also been found to be a highly effective preventive antioxidant and free radical scavenger, protecting organism from oxidative damage. In all species examined, plasma melatonin concentration shows a diurnal rhythm, with the highest levels during the night. Our studies are focused on the cues affecting Mel synthesis capacity in wild and farmed fish. The Mel rhythm in fish can be disrupted by fluctuations in surrounding calcium concentration or by prolonged thyroxine (T4) exposure. Physiological depletion of plasma ionized calcium may limit the capacity of Mel night production in two fish species, farmed rainbow trout and wild flounder. Prolonged (2 week) exposure to high level of exogenous thyroxine (T4) results in reduced night‐time plasma Mel levels, and may thus inhibit the melatonin‐related time‐keeping system in juvenile salmon. Disruption of the daily Mel rhythm implies a reduced ability of organisms to respond properly to environmental signals, and may be a useful indicator of disturbance in physiological functions.     

Heart rate responses to predation risk in Salmo trutta are affected by the rearing environment

Both wild‐ and hatchery‐reared brown trout Salmo trutta , 18 months of age and of the same genetic origin, responded with increased heart rates (tachycardia) to a simulated predator attack on 2 consecutive days. Brown trout reared in the hatchery showed a more rapidly induced tachycardia compared with wild‐reared fish at day 1, but not day 2. During an undisturbed period several hours after attacks, hatchery‐reared brown trout maintained higher heart rates compared to wild‐reared fish on both days. Behavioural responses to the attack were very low for all fish, although hatchery‐reared fish tended to be more active than wild fish after the attack day 2. The observed differences may have had a genetic background caused by different selection regimes in the hatchery‐ and wild‐rearing environments, or could have been due to different phenotypic responses in the two environments.     

The relationship between dominance behaviour and growth as a function of habitat stability and community complexity: a test using salmon and trout

Predation shortly after release is the main source of mortality among hatchery‐reared fish used to restore or enhance endangered salmonid populations. We found, that hatchery‐reared salmonid young originating from endangered stocks have weak innate responses to their natural fish predators. The ability to avoid predation in fish can be improved through social learning from experienced to naïve individuals. Huge benefits would be achieved, if social learning processes could be successfully applied on a large scale to enhance viability of hatchery fish prior to release into the wild. By using model predators together with chemical cues from real predators we tested if social learning could be used to train hatchery‐reared salmonid young to avoid fish predators. As there are clear differences in social behaviour among the salmonid species, we first examined whether these differences affect the probability and efficiency of learning anti‐predator skills from trained demonstrators. We compared anti‐predator responses of observers (fish trained by using experienced fish as demonstrators) with those of control fish, which had been 'trained' by untrained naïve conspecifics. We also examined how the efficiency of social learning depends on the ratio of experienced to naïve fish involved in social transmission trials. The results of these experiments will give guidelines how social learning could be utilized in developing hatchery scale training protocols.     

Domestication, comparative biology and interactions of wild and cultured fish: convenor's synthesis

Aquaculture is expanding rapidly and many fish species are brought into cultivation, entering a process of domestication with consequences for their morphology, physiology, ecology and evolution. In some species the abundance of cultured populations matches or exceeds that of wild stocks, and interactions between cultured and wild fish can pose significant conservation challenges. At the same time, captive breeding and re‐introduction play an important role in the conservation of some of the world's most endangered fishes. Drawing on contributions from the FSBI Symposium and the wider literature, we synthesize current knowledge of the process and extend of fish domestication, interactions between cultured and wild fish, and the use of cultured fish in fisheries enhancement and restoration. We provide a perspective on the role of biological issues within the wider context of aquaculture development and aquatic conservation biology, and conclude with a discussion of promising avenues for further research.     

Winter energy allocation and deficit of juvenile walleye pollock <Emphasis Type="Italic">Theragra chalcogramma</Emphasis> in the Doto area,northern Japan

Seasonal energy allocation and deficits of marine juvenile fishes have considerable effects on their survival. To explore the winter survival mechanism of marine fishes with low lipid reserves in their early life, juvenile walleye pollock Theragra chalcogramma were collected along the continental shelf of northern Japan over a 2-year period, and energy allocation and deficit patterns were compared between wild and laboratory-starved fish. Contrary to expectations, wild fish generally continued to accumulate protein mass and concurrently tended to reduce lipid mass from late autumn through winter. The most plausible explanation for the continuous structural growth is that juvenile pollock give priority to reducing mortality risk from size-selective predators under quasi-prey-limited conditions. Exceptionally, inshore small fish reduced both constituents during a winter. The inshore fish consumed 2.5 times more lipid energy than protein energy in November–December, but protein was more important than lipids as a source of energy in December–January and in February–March. However, dependence upon protein reserves was lower for the wild fish than for the laboratory-starved fish, suggesting milder nutritional stress of the wild fish than that observed in the starvation experiment. Moreover, the lipid contents of mortalities in the starvation experiment were mostly <1%, whereas few wild fish had such lipid contents in the field. These results suggest that juvenile pollock are able to avoid both starvation and predation by accumulating protein reserves.     

Stock and parental effects on embryonic and early larval development of winter flounder Pseudopleuronectes americanus (Walbaum)

A hierarchical breeding design was used to determine if winter flounder Pseudopleuronectes americanus embryos and yolk-sac larvae sired by Georges Bank males developed and grew larger than fish sired by Passamaquoddy Bay males, and to examine parental contributions to variations in fertilization success, time to 50% hatch, hatch success and larval morphological development. Significant stock effects were detected for time to hatch and larval development. Eggs fertilized by Passamaquoddy Bay males reached 50% hatch significantly earlier than eggs fertilized by Georges Bank males. Larvae sired by Georges Bank males were significantly larger during larval development for four of the six traits measured at 12 days post-hatch: head depth, jaw length, myotome height and body area. Embryo and larval development were strongly influenced by maternal contributions; there were significant maternal variance components for the majority of the variables measured. Paternal variance components were significant for fertilization success, time to hatch, larval jaw length and larval head depth, however, they acted principally through parental interactions. This information has important implications for the long-term sustainable development of winter flounder for aquaculture purposes as well as for understanding winter flounder genetic variation in the wild.     

Density-dependent predation and implications for stock enhancement with Japanese flounder

In laboratory predation trials kinsen-gani crab Matuta lunaris rarely preyed on hatchery-reared (HR) Japanese flounder Paralichthys olivaceus , even at extremely elevated prey densities ( c . 90 fish m⁻²), but fed readily on sacrificed Japanese founder. This suggests that under similar conditions in the wild predator-prey interactions between kinsen-gani and juvenile Japanese flounder are likely to be weak. Larger Japanese flounder readily cannibalized smaller conspecifics, suggesting that within-release group cannibalism may contribute considerably to post-release mortality of HR Japanese flounder when sufficient size variation exists within groups of released fish. Kinsen-gani presence had no effect on within-release group cannibalism rates of large Japanese flounder on small Japanese flounder. Within-release group cannibalism rates were significantly affected by densities of smaller (prey) fish, implying that the number of fish released (and thus post-release densities) in stock enhancement programmes may significantly affect post-release mortality rates. The probability of within-release group cannibalism increased rapidly as the cannibal-to-prey size ratio exceeded 1.6, suggesting that minimizing size variation of released HR individuals will reduce post-release cannibalism rates. These results have implications for Japanese flounder stock enhancement release strategies, and may be applicable to other finfish stock enhancement programmes.     

Interactions between endangered wild and hatchery salmonids: can the pitfalls of artificial propagation be avoided in small coastal streams?

Hatchery and wild juvenile populations of steelhead Oncorhynchus mykiss and coho salmon Oncorhynchus kisutch , in a small coastal watershed in central California, were sampled throughout the year in a stream and at a hatchery. Both species grew faster in captivity than in the wild. Hatchery fish of both species had elevated gill Na⁺, K⁺‐ATPase activity, and thus were ready to enter sea water when planted during the wild fish migration. Downstream migrant trapping and stream surveys indicated that hatchery smolts went to sea soon after planting, consequently avoiding the effects of competition and predation that commonly occur when hatchery‐bred juveniles are released. Adult steelhead were also sampled throughout the watershed. The return of hatchery steelhead was highly synchronized with that of wild steelhead, indicating that hatchery propagation had no adverse effects on the timing of the run. A disproportionate number of hatchery steelhead returned to the tributary where the hatchery was located, despite being planted throughout the watershed. Hatchery steelhead did not differ in mean age or size from wild steelhead. Observations of spawning indicated that hatchery and wild steelhead interbreed. Competition for mates or spawning substratum was rarely observed between hatchery and wild steelhead. Many of the problems commonly associated with artificial propagation can be avoided in small coastal watersheds when wild broodstock are used and fish are released as smolts.