An RNA:DNA-based growth model for young-of-the-year winter flounder Pseudopleuronectes americanus (Walbaum)

A laboratory calibration experiment was conducted to determine the relationship between nucleic acid-based variables and growth rate in young-of-the-year winter flounder Pseudopleuronectes americanus . Three temperatures and three feeding levels were used to produce a variety of growth rates. Nucleic acid analyses were conducted on white muscle tissue using an ultraviolet absorption assay. RNA concentration (μg mg⁻¹ wet tissue mass) and the ratio of RNA:DNA ( R _RD) were positively correlated with a mass-based instantaneous growth coefficient ( G _M) ( r = 0·42 and 0·72, respectively). Fifty-one per cent of the variability in growth rate was explained by the simple linear regression G _M=−0·02615 + 0·00848 R _RD ( P < 0·001). This model can be used to estimate recent growth rates for early juvenile winter flounder (27–52 mm standard length) at temperatures ranging from 11 to 24° C.     

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.     

Individual variability in the epidermis of prespawning inshore winter flounder, Pseudopleuronectes americanus

Prespawning winter flounder, Pseudopleuronectes americanus Walbaum, manifest epidermal thickening as a secondary sexual characteristic, but with considerable individual variability. The correlation between epidermal thickness and fish size in post-spawned flounder is less apparent in prespawning flounders due to factors associated with sexual maturation. Thus, epidermal thickness in individual prespawning flounder displays significant correlation with gonadosomatic index and. in females, with the degree of final maturation of vitellogenic oocytes. However, both epidermal thickness and gonadosomatic index of individual flounders also display significant correlation with condition factor. It is proposed that body size and condition factor represent important primary physiological determinants influencing individual variability in seasonal epidermal thickness, and that factors associated with sexual maturation represent a secondary physiological determinant.     

Seasonal prevalence of the microsporidan, Glugea stephani (Hagenmuller), in winter flounder, Pseudopleuronectes americanus (Walbaum), from the New York-New Jersey Lower Bay Complex

During a 32 month period 26 monthly collections of winter flounder were conducted from various locations in the New York-New Jersey Lower Bay Complex. A total of 3125 flounder were captured and examined for the presence of the microsporidan, Glugea stephani. Of the total number of fish collected, 260 (8.32%) were infected with the protozoan. At least one G. stephani infected flounder was captured and identified each month, which indicates that the infection is present on a year-round basis. The monthly infection prevalence ranged from 0.63 to 25%. Increased parasitism corresponded with elevated water temperatures. Fish size was not a statistically significant factor for infection.     

Ion transport across the isolated intestinal mucosa of the winter flounder,Pseudopleuronectes americanus

The isolated intestinal mucosa of the flounder, Pseudopleuronectes americanus, when bathed in a 20 mM HCO3-Ringer's solution bubbled with 1% CO2 in O2, generated a serosa-negative PD and, when short-circuited, absorbed Cl at almost 3 times the rate of Na. Reducing HCO3 to 5 mM decreased the net Cl flux by more than 60%. The following results suggest that, despite the PD, Na and Cl transport processes are nonelectrically coupled: replacing all Na with choline abolished both the PD and net Cl flux; replacing all Cl with SO4 and mannitol abolished the PD and the net Na flux; and adding ouabain (to 0.5 mM) abolished the PD and the net Cl flux. Nearly all of the unidirectional serosa-to-mucosa Cl flux (JClsm) seemed to be paracellular since it varied with PD and Cl concentration in a manner consistent with simple diffusion. JClsm was only about one-fourth of JNasm, suggesting that the paracellular pathway is highly cation-selective. The data can be explained by the following model: (i) Na and Cl uptake across the brush border are coupled 1 : 1; Na is pumped into the lateral space and Cl follows passively, elevating the salt concentration there; (ii) the tight junction is permeable to Na but relatively impermeable to Cl; and (iii) resistance to Na diffusion is greater in the lateral space (considered in its entirety) than in the tight junction. If these assumptions are correct, the serosa-negative transmural PD is due mainly to a salt diffusion potential across the tight junction and, under short-circuit condition, most of the Na pumped into the lateral space diffuses back into the luminal solution, whereas most of the Cl enters the serosal solution. Morphological features of the epithelium support this interpretation: the cells are unusually long (60 micrometer); there is little distension of the apical 12 micrometer of the lateral space during active fluid absorption; and distension distal to this region is intermittently constricted by desmosomes.     

Thermoregulatory behavior and diel activity of yearling winter flounder,Pseudopleuronectes americanus (Walbaum)

Synopsis Sixteen yearling winter flounder, tested individually for 3-day periods in electronic shuttleboxes, voluntarily occupied an 8–27°C range of temperature, with a modal final thermal preferendum of 18.5°C (mean 18.7°C, median 19°C, midpoint 17.5°C, s.d. 1.9°C, S _k + 0.33). The locomotor activity pattern of the yearling fish in the laboratory was markedly nocturnal, with mean hourly nocturnal activity exceeding mean hourly diurnal activity by a factor of 3.4. Maximum activity occurred at 0300 EST, minimum activity at 1400 EST. While activity generally increased with temperature, a local activity minimum occurred at 18.7°C, coinciding with the 24-hour mean final thermal preferendum. Comparison of these laboratory data with previously published field data suggest that behavioral responses to temperature and light play significant roles in determining age- and size-specific differences in seasonal depth stratification and onshore-offshore distributions in this species.     

Post-fertilization changes in the chorion of winter flounder, Pseudopleuronectes americanus Walbaum, eggs observed with scanning electron microscopy

Little has been reported on the fine structure of the outer membrane of fish eggs during and after fertilization. When observed in the scanning electron microscope, the unfertilized egg of the winter flounder, Pseudopleuronectes americanus , is characterized by a crisscross pattern of depressions. These depressions radiate in all directions across the membrane surface creating a wrinkled appearance. After fertilization, the surface of the chorion becomes regular with a smoother appearance. The pores of the unfertilized egg are flush with the chorion surface, but become thickened and elevated after fertilization. While the chorion of the unfertilized egg is also smooth and uniformly textured, the chorion of the fertilized egg appears granular by first cleavage of the blastodisc. Although no apparent change occurs in the distance between pores after fertilization, statistically significant decreases in pore diameter occur 5 min after fertilization. These results are compared to those on egg membranes of other species of fish and invertebrates.     

The binding of zinc to the soluble proteins of intestinal mucosa in winter flounder (Pseudopleuronectes americanus).

1. The binding of Zn2+ to soluble proteins of intestinal mucosa of winter flounder was examined using an equilibrium dialysis technique. 2. There appeared to be more than one binding system present for Zn2+ in the mucosal cytosol. 3. It required four times the normal endogenous Zn2+ level found in the mucosal cytosol to saturate the highest affinity (K1 = 2.42 x 10(7] binding system. 4. Of 10 metals tested Cu2+ was the only one which interfered with Zn2+ binding to the mucosal cytosol proteins. 5. It is postulated that binding proteins in the mucosal cytosol of winter flounder may play a role in the transport of Zn2+.     

Taxonomy and host-specificity of Gyrodactylus aideni n. sp. and G. pleuronecti (Monogenea: Gyrodactylidae) from Pseudopleuronectes americanus (Walbaum) in Passamaquoddy Bay, New Brunswick, Canada

Wild and cultured winter flounder Pseudopleuronectes americanus (Walbaum) from Passamaquoddy Bay were surveyed for species of Gyrodactylus Nordmann, 1832. Two species were found: G. pleuronecti Cone, 1981 and G. aideni n. sp, both members of Malmberg’s ‘groenlandicus group’. Although the hard parts in the haptor are very similar in the two species, hamuli of G. aideni are consistently shorter than those of G. pleuronecti. The two species differed by 35 base pairs in the ITS 1, 5.8 and ITS 2 region. A BLAST search identified a variety of species of Gyrodactylus from marine fishes in the Atlantic Ocean as closest matches, indicating the ‘groenlandicus group’ is part of a major marine lineage within Gyrodactylus (sensu lato) that has successfully radiated among coastal percid, pleuronectid, cottid and anarhichadid fishes. Exposure experiments suggested that winter flounder is the primary host of both species of parasites and that three other pleuronectid species in the bay may potentially serve only as occasional transport hosts.     

Epidermal condition in post-spawned winter flounder, Pseudopleuronectes americanus (Walbaum), maintained in the laboratory and after exposure to crude petroleum

Skin samples were taken, during the post-spawning period, from Pseudopleuronectes americanus maintained in both stationary and in running-water systems, and after exposure to Venezuelan crude oil in each system. Comparisons with skin taken freshly from an inshore population revealed that dissociation of epidermal tissue, epidermal thinning and reduced epidermal melanophore frequency normally associated with the postspawning period were not so apparent after laboratory confinement and were even less pronounced after exposure to crude oil. In post-spawned, laboratory maintained flounders, there was also correlation between epidermal thickness and the weight of the fish. Indications from the stationary system suggest that stress associated with environmental conditions may evoke mucigenesis in the superficial epidermal cells as distinct from the goblet cells.     

The effect of androgen treatment on the epidermis of post-spawned winter flounder, Pseudopleuronectes americanus (Walbaum)

The effect of androgen treatment on the thin epidermis of post-spawned winter flounder, Pseudopleuronectes americanus (Walbaum), freshly collected from an inshore population was investigated. Over the duration of the experiment, control flounders displayed epidermal thickening and increased goblet cell frequencies associated with captivity in this species. Treatment of males with 11-ketotestosterone induced significantly more differential thickening of the blindside epidermis. Compared with the controls testosterone treatment did not induce additional thickening of male epidermis, but it did increase differential thickening on the blind side of females.     

Ion transport across the isolated intestinal mucosa of the winter flounder,Pseudopleuronectes americanus: II. Effects of cyclic AMP

Summary Addition of cyclic AMP and theophylline to the intestinal mucosa of the winter flounder,Pseudopleuronectes americanus decreased short-circuit current and net Na and Cl absorption and increased total conductance and the serosa-to-mucosa unidirectional Cl flux (J _sm ^Cl ). The last two changes were independent of the original rate of NaCl absorption and persisted even when net absorption of Na and Cl had been abolished by ouabain. Voltageclamp experiments revealed that the increment inJ _sm ^Emphasis>/Cl produced by cyclic AMP is PD-insensitive and therefore not due to an increase in the Cl conductance of the paracellular shunt. Cyclic AMP appears, therefore, both to inhibit net NaCl absorption and to increase the Cl permeability and total conductance of the intestinal epithelial cells; its failure to stimulate secretion (in contrast to its action on mammalian intestine) may be related to the absence of crypts in flounder intestinal epithelium.     

Seasonal patterns of winter flounder Pseudopleuronectes americanus abundance and reproductive condition on the New York Bight continental shelf

To resolve varied and sometimes conflicting accounts of spawning and habitat characteristics for winter flounder Pseudopleuronectes americanus, seasonal patterns in abundance and reproductive condition were investigated in the New York Bight, near the southern edge of their current reproductive range. Fish were collected from trawl surveys on the inner continental shelf from October 2006 to October 2007. Pseudopleuronectes americanus were most abundant during January and April surveys, were rarely collected in August, with intermediate abundances in June and October. Measurements of fish condition [hepato‐somatic index (I_H), condition factor (K) and the per cent dry mass of muscle tissue (%M_D)] and reproductive condition [gonado‐somatic index (I_G)] were determined to evaluate seasonal changes in energy accumulation and depletion and reproduction. Males and females had similar patterns in body and reproductive condition, although the magnitude of change was greater for females. I_H values were highest during spring and early summer, suggesting increased feeding following spawning. K and %M_D increased through spring and summer then declined in the autumn and winter concurrent with gonadal development. Gonads began developing in the autumn, and in January, I_G values approached spawning levels, with many spent individuals collected in spring. Within these general patterns, however, there was a large degree of variability among individuals, and a few mature non‐reproductive (‘skipped spawning’) females were observed. In the period after spawning, increased energy intake, indicated by increased I_H, may influence reproductive output since this energy is gradually transferred to the muscle and used for gonadal development in the forthcoming year. The occurrence of ripening individuals on the inner continental shelf in January suggests that these fish either rapidly move into estuaries to spawn by February–March or they remain on the inner shelf to spawn, or some combination of these. Future studies should evaluate these possibilities, as both estuarine and inner shelf habitats are potentially affected by activities such as dredging, sand dredging and wind energy development.     

Epithelial cell proliferation in the intestine of the winter flounder,Pseudopleuronectes americanus

Summary To study epithelial cell proliferation in the North American flounder (Pseudopleuronectes americanus), fed and fasted fish received intravenous injections of ³H-thymidine and were killed 11/2 to 2 h later. Radioautographs of proximal, middle, and distal intestinal segments revealed proliferating epithelial cells at all levels of intestinal folds including the crest although labelled nuclei were most abundant in the epithelial cells on the lower half of folds and between folds. Mature appearing goblet cells with labelled nuclei were observed at all levels of the folds. The mean labelling index was greater in the epithelium of fed than fasted flounder. In fed flounder the mean labelling index was greatest in the proximal segment and least in the distal segment; no substantive differences in mean labelling indices were observed in the various segments of intestine from fasted fish. Electron microscopy revealed no major structural differences among epithelial cells along the base of folds compared to cells near the crest of folds. These findings indicate that 1) epithelial cell proliferation occurs at all levels of the folds of flounder intestine and is not compartmentalized to the base of the folds and interfold epithelium as reported in other teleosts, and 2) epithelial cell proliferation in the flounder intestine varies with feeding status.Supported be research grants AM 17537 and RR 05764 from the National Institutes of Health, Bethesda, Maryland and grant DEB7826821 AO1 from the National Science Foundation, Washington, D.C.The authors are grateful to Dr. Michael Field for stimulating discussions and suggestions and for providing facilities for collecting material from fish     

Adaptation of renal function to hypotonic medium in the winter flounder (Pseudopleuronectes americanus)

Summary The kidneys of winter flounders transferred to hypotonic medium were investigated for glomerular and tubular handling of fluid and electrolytes and for the urinary excretion of proteins. Media were sea water (925 mosm·kg^–1) and brackish water (70 mosm·kg^–1).In sea water, the urine was hypertonic to the plasma in 7 fish of this study. Urine flow rate was correlated with the GFR. After adaptation to brackish water a delay of 1 to 3 days was observed until the kidneys switched from fluid retention to the excretion of large amounts of dilute urine. GFR and urine flow rate were increased from 0.61±0.08 to 1.58±0.29 ml·h^–1·kg^–1 and from 0.14±0.02 to 0.68±0.08 ml·h^–1·kg^–1, respectively . With increased filtered load the tubular reabsorption of fluid decreased from 74±2.4% to 45±11.2%. The excretion rates of sodium and potassium were increased due to decreased fractional sodium and potassium reabsorption. The urinary excretion of divalent cations, however, was reduced because the net tubular reabsorption of calcium was increased and the net secretion of magnesium was diminished.Both the urinary total protein concentration and the protein pattern showed no significant change, but the rate of protein excretion was increased from 0.21±0.04 to 0.60±0.05 mg·h^–1·kg^–1. The comparison of protein patterns obtained from urine and serum samples revealed that high molecular weight (HMW) proteins prevail in the serum whereas low molecular weight (LMW) proteins dominate in the urine. The diminished quantity of the HMW-protein fraction in the urine thus may reflect size selectivity of the glomerular filtration barrier for serum proteins also in the winter flounder.Abbreviations BW brackish water - SW sea water - GFR glomerular filtration rate - HMW heigh molecular weight - LMW low molecular weight     

A comparative histological and histochemical study of the post-gastric alimentary canal from three species of pleuronectid, the Atlantic halibut, the yellowtail flounder and the winter flounder

The histology and mucus histochemistry of the pleuronectid post-gastric alimentary canal was examined using light and electron microscopy. Distinct differences in goblet cell mucus histochemistry were observed between species, with the two closest taxonomic species, the winter flounder and the yellowtail flounder showing the most diversity and the halibut showing regional variation. Numbers of goblet cells within post-gastric regions did not differ significantly between species, but were significantly different between regions within species increasing toward the rectum. The post-gastric region was divisible into two areas based upon the ultrastructural features of lipid digestion and absorption in the intestine and pyloric caeca, and of exogenous protein in the rectum. The combination of species-specific histochemical differences in mucus and general histological and ultrastructural differences within the post-gastric regions between these species suggest a correlation between lumenal environmental conditions/histology and natural prey preference.