Feeding behavior of newly settled winter flounder (Pseudopleuronectes americanus) on calanoid copepods

Field and laboratory investigations were conducted to examine feeding by newly settled winter flounder (Pseudopleuronectes americanus) on two co-occurring calanoid copepods, Eurytemora affinis and Acartia hudsonica. During the spring, these prey are present when winter flounder initiate their demersal lifestyle in estuaries of the northeastern United States. Epibenthic zooplankton were collected concurrently with winter flounder in the Navesink River estuary, NJ, in May 1998 and 1999. Although both calanoid species were in the estuary during the 2-year survey, E. affinis was consumed nearly to the exclusion of A. hudsonica by newly settled winter flounder. Annually, E. affinis and A. hudsonica had similar size distributions in field collections, indicating that species choice was not size selective. However, when preying on E. affinis, winter flounder preferred the larger sized organisms. In single species laboratory experiments, E. affinis and A. hudsonica were consumed equally by newly settled winter flounder (19-23 mm TL), but there were more strikes made toward E. affinis. Despite the lower catch efficiency, E. affinis was selected over A. hudsonica when the prey species were offered together in equal numbers. The selection for E. affinis over A. hudsonica by newly settled winter flounder may be the result of behavioral and/or morphological differences in the prey species.     

Differential amplification of antifreeze protein genes in the pleuronectinae

Summary The organization of antifreeze protein (AFP) genes in the yellowtail flounder was investigated by Southern blotting and the characterization of clones from a genomic library. This flounder, like the closely related winter flounder, has a set of 10–12 linked but irregularly spaced AFP genes. However, it lacks the tandemly amplified set of 20 such genes that are present in the winter flounder. DNA sequence analysis of a tandemly repeated gene from winter flounder showed that it can code for one of the two most abundant AFP components in the serum. Consistent with this higher AFP gene dosage, the peak serum AFP level in midwinter was 9 mg/ml in the winter flounder and only 4 mg/ml in the yellowtail flounder. A recent amplification of the AFP gene in the winter flounder lineage might be responsible for the higher serum AFP levels in this fish. This increase in gene dosage might have helped the winter flounder colonize the ice-laden, shallow-water niche that it currently occupies along the east coast of North America. Genomic Southern blotting of two other righteye flounders, the smooth flounder and the American plaice, illustrates another example of a differential amplification of AFP genes that correlates with a species' exposure to ice.     

Distribution and ontogeny of digestive enzymes in larval yellowtail and winter flounder

The distribution of digestive enzymes was studied using histoenzymological methods in yellowtail and winter flounder larvae reared on three different diets: live food, weaned at day 15, or starved. Alkaline phosphatase, dipeptidyl peptidase IV, aminopeptidase M and esterase were present at 3 days post-hatch and became differentially distributed coinciding with morphological development. For larvae fed a live diet, activity of these enzymes was present in the intestine of both species and rectum of yellowtail flounder. Alkaline phosphatase was also present in the post-oesophageal swelling (stomach anlage) of winter flounder. Activity of all enzymes was absent in starved winter flounder larvae and a decrease in aminopeptidase M and esterase activity occurred in starved yellowtail flounder larvae. Acid phosphatase was not identified in either species. The eVect of weaning on enzymatic activity was not evaluated fully as the larvae did not survive long enough after the introduction of the artificial diet to complete experimentation.     

Size-related shifts in the habitat associations of young-of-the-year winter flounder (Pseudopleuronectes americanus): field observations and laboratory experiments with sediments and prey

Field surveys and laboratory studies were used to determine the role of substrata in habitat selection by young-of-the year winter flounder. A synoptic field survey of winter flounder and sediments in the Navesink River-Sandy Hook Bay estuarine system in New Jersey demonstrated that winter flounder distribution was related to sediment grain size. Analysis using a generalized additive model indicated that the probability of capturing 10-49 mm SL winter flounder was high on sediments with a mean grain diameter of /=40 mm SL) preferred coarse-grained sediments. Burying ability increased with size and all flounders avoided sediments that prevented burial. Subsequent laboratory experiments revealed that the presence of live prey (Mya arenaria) can over-ride sediment choice by winter flounder (50-68 mm SL) indicating the complexity of interrelated factors in habitat choice.     

Predator-prey relations between age-1+ summer flounder (Paralichthys dentatus, Linnaeus) and age-0 winter flounder (Pseudopleuronectes americanus, Walbaum): predator diets, prey selection, and effects of sediments and macrophytes

Laboratory experiments and weekly trammel net surveys in the Navesink River, New Jersey (USA) were used to examine the predator-prey interaction between age-1+ summer flounder (Paralichthys dentatus) and age-0 winter flounder (Pseudopleuronectes americanus). Winter flounder (24-67 mm TL) were the dominant piscine prey of summer flounder (n=95, 252-648 mm TL) collected in trammel nets. We observed a temporal shift in summer flounder diets from sand shrimp (Crangon septemspinosa) and winter flounder, dominant during June and early July, to blue crabs (Callinectes sapidus) and other fishes (primarily Atlantic silversides, Menidia menidia and Atlantic menhaden, Brevortia tyrannus) later in the summer. Variations in prey selection appeared to be related to changes in the spatial distribution of predators and spatio-temporal variation in prey availability. In laboratory experiments, summer flounder (271-345 mm total length, TL) preferred demersal winter flounder to a pelagic fish (Atlantic silversides) and a benthic invertebrate (sand shrimp) prey, and the vulnerability of winter flounder increased with increasing prey body size from 20 to 90 mm TL. Experiments testing habitat effects showed that mortality of winter flounder in three different size classes (20-29, 40-49, 60-69 mm TL) was not influenced by sediment grain sizes permitting differential burial of the prey. However, vegetation enhanced survival, with fish suffering lower mortality in eelgrass (Zostera marina, 15+/-0.04%) than in sea lettuce (Ulva lactuca, 38+/-0.04%) or bare sand (70+/-0.07%) when the macrophytes were planted to produce similar leaf surface areas (5000 cm(2) m(-2)). Prey vulnerability appeared to be related to the role of vision in the predator's attack strategy and prey activity levels.     

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.     

Factors affecting the post‐release survival of cultured juvenile Pseudopleuronectes americanus

Laboratory experiments were performed with cultured and wild juvenile winter flounder Pseudopleuronectes americanus to evaluate differences in behaviour and adaptation affecting post‐release vulnerability to predation. Studies revealed that the cryptic abilities of cultured winter flounder increased over time. Sediment‐naïve, cultured fish required a minimum of 2 days to improve their burying skills and at least 90 days for colour adaptation to match the sediment. Cultured winter flounder selected sediments consisting of small grains and colours matching their own pigment. Cultured winter flounder, regardless of their colour, were significantly more vulnerable to predation by birds. Additionally, cultured fish reacted differently than wild winter flounder when exposed to cues from a potential predator.     

Predatory impact of the green crab (Carcinus maenas Linnaeus) on post-settlement winter flounder (Pseudopleuronectes americanus Walbaum) as revealed by immunological dietary analysis

Predation on flatfish during the early juvenile stage is an important factor regulating year-class strength and recruitment. In this study, immunological dietary analysis was performed on green crabs (Carcinus maenas) collected from the Niantic River, Connecticut, in an effort to evaluate the predatory impact of this species on post-settlement winter flounder (Pseudopleuronectes americanus). Through the use of species-specific antiserum, winter flounder proteins were identified in 4.8% of the green crab stomachs analyzed (n = 313, size range = 14-74 mm carapace width, CW), revealing that crabs ≥ 29 mm CW are predators of post-settlement winter flounder in natural populations. The most significant factor underlying the predator-prey interaction was the relative size relationship between species, such that the incidence of winter flounder remains in the stomach contents of green crabs was positively correlated with predator-to-prey size ratio. Results from dietary analysis were incorporated into a deterministic model to estimate the average daily instantaneous mortality and cumulative mortality of winter flounder owing to green crab predation. Accordingly, green crabs may account for 0.4% to 7.7% (mean = 2.2%) of the daily mortality of winter flounder and consume 1.1% to 32.3% (mean = 10.2%) of the flounder year-class. Model simulations further indicate that variations in green crab abundance and size-structure account for the greatest variability in winter flounder mortality. Relative to other macro-crustacean predators, however, predation by green crabs has a minimal effect on winter flounder survival, due in large part to the low densities of these crabs in temperate estuaries.     

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.     

A natural variant of type I antifreeze protein with four ice-binding repeats is a particularly potent antifreeze.

A 4.3-kDa variant of Type I antifreeze protein (AFP9) was purified from winter flounder serum by size exclusion chromatography and reversed-phase HPLC. By the criteria of mass, amino acid composition, and N-terminal sequences of tryptic peptides, this variant is the posttranslationally modified product of the previously characterized AFP gene 21a. It has 52 amino acids and contains four 11-amino acid repeats, one more than the major serum AFP components. The larger protein is completely alpha-helical at 0 degree C, with a melting temperature of 18 degrees C. It is considerably more active as an antifreeze than the three-repeat winter flounder AFP and the four-repeat yellowtail flounder AFP, both on a molar and a mg/mL basis. Several structural features of the four-repeat winter flounder AFP, including its larger size, additional ice-binding residues, and differences in ice-binding motifs might contribute to its greater activity. Its abundance in flounder serum, together with its potency as an antifreeze, suggest that AFP9 makes a significant contribution to the overall freezing point depression of the host.     

Isolation, characterization and seasonal variations in the concentration of N-epsilon-(gamma-glutamyl)-lysine isodipeptide in the blood plasma of the winter flounder (Pseudopleuronectes americanus).

A ninhydrin-positive compound was isolated from the plasma of the winter flounder (Pseudopleuronectes americanus) and identified as the isodipeptide N-epsilon-(gamma-glutamyl)-lysine. Wide seasonal variations in plasma concentration of this compound are found in the male flounder, whereas it is present only in trace amounts in the plasma of the female flounder throughout the year. In the male flounder the plasma concentration of N-epsilon-(gamma-glutamyl)-lysine rises from January to a peak in May and June. It is during these latter months that the flounder spawn. After spawning, the plasma concentration of the isodipeptide decreases to low basal values in October. These changes in plasma concentration of N-epsilon-(gamma-glutamyl)-lysine may be related to changes in testes weight, spermatogenesis and spawning activity of the winter flounder.     

Epidermal characteristics of reproductive subsets in an inshore population of winter flounder, Pseudopleuronectes americanus

The winter flounder, Pseudopleuronectes americanus (Walbaum), population off Newfoundland can include a 'non-reproductive post-mature' subset at any time together with 'reproductive' and 'immature' subsets. Skin samples were taken from male and female specimens from each subset in the winter, after reproductive flounder have normally undergone a pre-winter phase of gonadal recrudescence, and also during the spring prespawning period and in the summer following spawning. Results for immature flounder demonstrate that fish size influences epidermal thickness in this species. The epidermis of reproductive flounders is thick, representing a secondary sexual characteristic, during gonadal recrudescence. A thick epidermis is most pronounced on the blind-side, particularly in prespawning males. Although the epidermis of non-reproductive post-mature fish was thinner than in prespawning males and females, it was thicker than in immature flounders. Also, the epidermal thickness of non-reproductive post-mature individuals can still display some seasonal change although gonadal recrudescence has not been sustained. There is also a relationship between condition factor and epidermal thickness which is manifested particularly clearly in immature and non-reproductive post-mature flounder.     

Comparison of routine oxygen consumption rates of three species of pleuronectids at three temperatures

The oxygen consumption rates of three species of pleuronectids, the yellowtail flounder, Pleuronectes ferrugineus (Storer), the winter flounder, Pleuronectes americanus (Walbaum), and the American plaice, Hippoglossoides platessoides (Fabricius), were examined under simulated, land-based, aquaculture conditions. Routine oxygen consumption (ROC) rates for groups of each species were measured simultaneously using single-pass, flow-through respirometry. This study was conducted over three seasons at temperatures from 2°C to 14°C. An analysis of variance identified a significant interaction between temperature and species on the oxygen consumption rates of these flounder. The analysis indicated that at each temperature, ROC rates were significantly different among the three species (P < 0.05). A subsequent test of each species'ROC rate across the three temperatures indicated that both the yellowtail flounder and the winter flounder had significantly different ROC rates at each temperature experiment (P < 0.05). The ROC of yellowtail and winter flounder responded similarly to changes in experimental temperatures.     

Developmental fate of the yolk protein lipovitellin in embryos and larvae of winter flounder, Pleuronectes americanus.

The developmental fate of the vitellogenin-derived yolk protein, lipovitellin (Lv), was investigated in winter flounder embryos and yolk-sac larvae. Since Lv is present as only one major polypeptide in ovulated winter flounder eggs, unlike the multiple yolk polypeptides found in the mature eggs of most teleosts, this system is presented as a simpler model of yolk protein structure and utilization during teleostean development. Winter flounder Lv is cleaved during embryogenesis from a 94 kD polypeptide at fertilization to 67 kD and 26 kD polypeptides at hatching. The rate of this proteolytic processing is slow during early embryonic development, but enters a more rapid phase between days 8 and 12 post-fertilization in embryos reared at 4-5 degrees C, and approaches 50% completion at day 10. Lv processing is essentially complete 3 days before hatching; nevertheless, major degradation of the Lv peptide by the developing winter flounder does not occur until after hatching. The Stokes radius of Lv changes only moderately following processing, from 4.50 nm in unfertilized eggs to 4.19 nm in late embryos and newly hatched larvae, whereas the processed Lv retains its heat stability relative to other yolk polypeptides. Nearly 50% of its lipid content, however, is released from the Lv particle during embryogenesis, concomitant with cleavage of the Lv 94 kD polypeptide. Lv processing may thus render a portion of the yolk protein-associated lipid more accessible to the developing embryo, whereas other yolk components are retained for later use by the winter flounder larva. Alternately, removal of lipid may lead to proteolytic vulnerability of the Lv polypeptide. In either case, only a portion of the lipid moiety of the Lv particle appears to play a significant nutritive role for the embryo, whereas its protein component is reserved for larval use. J. Exp. Zool. 284:686-695, 1999.     

Effect of sediment,salinity, and velocity on the behavior of juvenile winter flounder (<Emphasis Type="Italic">Pseudopleuronectes americanus</Emphasis>)

Winter flounder (Pseudopleuronectes americanus) is a benthic flatfish that is economically important for recreational and commercial fishing in North America. In the last twenty years, the species has undergone a drastic decline, mainly due to anthropic influence. The goal of this study was to gain knowledge on habitat preferences and behavior of juvenile winter flounder to improve the management of natural stocks and optimize release sites of juveniles produced for stock enhancement. Three abiotic factors (sediment, current, and salinity) potentially influencing the distribution of flatfish species were tested in a recircurlating flume with juvenile winter flounder. Time budgets of observed behaviors including swimming, orientation, and burying capacity were analyzed. Sediment texture was the only factor that significantly influenced the burying behavior of winter flounder juveniles; shear velocity, salinity, and sediment had no effect on the orientation of juveniles.     

Ontogenetic variation in habitat associations for four flatfish species in the Gulf of Maine-Georges Bank region

Multivariate ordination techniques were used to examine how size classes of four flatfish species, American plaice Hippoglossoides platessoides , winter flounder Pseudopleuronectes americanus , yellowtail flounder Limanda ferruginea and fourspot flounder Paralichthys oblongus , are related to bottom depth, bottom temperature, substratum grain size and temporal factors using a 35 year time series from autumn and spring in the Gulf of Maine-Georges Bank region. Depth had the most explanatory value during both seasons in most cases, with fish size generally increasing with depth. One exception was yellowtail flounder in the spring for which a temporal factor explained the most variance, reflecting an increase in size over the time series due to changes in fishing pressure. Temperature was secondarily important for yellowtail flounder in the autumn and for fourspot flounder in both seasons. Substratum type was secondarily important for winter and yellowtail flounders in the spring with smaller fish associated with larger substratum types. Seasonal associations with depth, temperature and substratum are related to seasonal spawning migrations, thermal preferences and other ecological factors.     

Localization of vitellogenin-related protein in the developing oocytes of winter flounder (Pleuronectes americanus) by protein A-gold immunocytochemistry

Vitellogenin-related proteins were localized within the oocytes of a marine teleost, the winter flounder, using protein A-gold immunocytochemical electron microscopy. A homologous vitellogenin antisera was used along with 15-nm protein-A gold on thin sections of oocytes. Localization was significantly confined to organelles described as yolk spheres within the oocyte cytoplasm. This technique provides specific identification of yolk-containing organelles within the developing winter flounder oocyte.     

Estuarine and habitat-related differences in growth rates of young-of-the-year winter flounder (Pseudopleuronectes americanus) and tautog (Tautoga onitis) in three northeastern US estuaries

Instantaneous growth rates of young-of-the-year winter flounder Pseudopleuronectes americanus (Walbaum) (12.0-60.4 mm standard length, SL) and tautog Tautoga onitis (Linnaeus) (21.4-73.8 mm total length, TL) from three estuarine systems in New Jersey (Great Bay-Little Egg Harbor and Navesink River) and Connecticut (Hammonasset River) were used in an attempt to assess the relative quality of selected nominal habitats. A series of short-term field caging experiments were conducted during 1994 and 1995 in: macroalgae (primarily, Ulva lactuca), eelgrass (Zostera marina), unvegetated areas adjacent to macroalgae and eelgrass and tidal creeks in Spartina dominated marsh. Growth rates varied with habitat, estuary and year. Comparisons across nominal habitats within and among estuaries did not show any one habitat with consistently higher growth, and growth was relatively independent of whether a habitat was vegetated or adjacent to vegetation. The growth rates of winter flounder and tautog from the Hammonasset River were not different among habitats in either year of the study. In the Great Bay-Little Egg Harbor, both winter flounder and tautog had higher growth rates in macroalgae with growth in eelgrass varying significantly between years. Conversely, in the Navesink River both species had higher growth rates in eelgrass. Environmental changes associated with temperature and dissolved oxygen appeared to influence growth rates. Winter flounder growth rate and survival was depressed in tidal marsh creeks in the three estuaries and in vegetated macroalgae habitats in the Navesink River where dissolved oxygen levels were often very low (<2 mgl(-1)) for extended periods. In summary, the growth rates of the young-of-the-year of these two species varied temporally and were dependent on the interaction of both the specific estuary and habitat in which the experiments took place. Further, habitat quality, as defined by relative growth rate, was difficult to evaluate because it can be variable and nominal habitat designations are often not sufficient to define the boundaries of a species habitat requirements.