Water balance in eggs of striped bass (Morone saxatilis)

Measurements of embryonic volume revealed regulation capability before hatch. Water diffusion permeability, determined as tritiated water efflux, was found to be low, but within the range for fish eggs. The perivitelline fluid is readily interchangeable with the external medium due to a high chorion permeability to both water and salt.     

An in vitro assay to measure phagocytosis in striped bass hybrids (Morone saxatilis x Morone chrysops)

An in vitro phagocytosis assay was developed for hybrid striped bass (Morone saxatilis x Morone chrysops), using cells collected from the peritoneal cavity of this fish. The findings indicated that: (1) 10 days following a single intraperitoneal injection (1 ml) of Freund's incomplete adjuvant (FIA) was an appropriate time for collecting suitable working concentrations (5.3+/-4.0 x 10(7) cells ml(-1)) of peritoneal phagocytes (83.7+/-1.5% macrophages) from these hybrids held at 23 degrees C; (2) these cells phagocytosed latex beads (polystyrene microspheres 3.12 microm in diameter) after 30 min of in vitro incubation at room temperature (25+/-1 degrees C). The phagocytic ability and phagocytic capacity in a washed adherent layer exposure system were 67.2+/-2.76% and 4.14+/-0.35 beads phagocyte(-1), respectively. These results strongly suggest that a simple methodology, including baseline data serving as guidelines, is now available for conducting in vitro phagocytosis assays in this hybrid.     

Blood oxygen transport in stressed striped bass (Morone saxatilis): role of beta-adrenergic responses

Summary Strenuous 5-min exercise resulted in a 0.3 unit drop in the dorsal aortic pH of striped bass. The acidosis was metabolic: the blood lactate concentration increased during the exercise, whereas blood CO₂ tension decreased. Dorsal aortic oxygen content was maintained despite the acidosis. This was a result of increased blood O₂ tension, haemoglobin concentration and red cell volume, decreased cellular nucleoside triphosphate (NTP) concentration, and decreased proton gradient across the red cell membrane. When the fish were treated with the beta-antagonist, propranolol, before the exercise, the arterial oxygen content decreased significantly in the stress. The mean cellular haemoglobin concentration and cellular NTP concentration increased slightly, and the proton gradient across the red cell membrane decreased less than in control exercise. These results show that the beta-adrenergic responses of striped bass red cells play an important role in maintaining the arterial O₂ content in stress.Abbreviation NTP nucleoside triphosphates     

Measurement of larval striped bass (Morone saxatilis) net avoidance using evasion radius estimation to improve estimates of abundance and mortality

Net avoidance rate increases as a function of larval striped bass size.This causes under-estimation of abundance and overestimation of mortalityrate. We modeled net avoidance by assuming that fish avoid the net byswimming a radial distance at a right angle to the net axis. This distance,the evasion radius, was estimated by comparing the calculated densities ofstriped bass larvae from a series of paired tows involving a large and asmall net. Iteration and solution models were used to estimate the evasionradius for each millimeter size group of fish in order to estimate theactual density in the environment. Avoidance of the nets increased withfish length. The ratio of actual density in the environment to the measureddensity in the small net was used to adjust abundances measured in ourichthyoplankton surveys. After adjusting for net avoidance, mortality ratesof striped bass larvae from the Sacramento-San Joaquin Estuary were reducedby 10% compared to the unadjusted rates.     

Tissue essential fatty acid composition and competitive response to dietary manipulations in white bass (Morone chrysops), striped bass (M. saxatilis) and hybrid striped bass (M. chrysopsxM. saxatilis)

The effects of wide changes in dietary levels of docosahexaenoic (DHA) or arachidonic (ArA) acids on growth, survival and fatty acid composition in body tissues of Morone larvae were examined. White bass (WB, Morone chrysops), striped bass (SB, Morone saxatilis) and sunshine hybrid bass (HSB, M. chrysopsxM. saxatilis) larvae (day 24-46) were fed Artemia nauplii enriched with algal sources of varying proportions of DHA and ArA (from 0 to over 20% of total fatty acids). WB larvae fed DHA-deficient Artemia diet retarded over 50% of their potential growth, however, increasing dietary DHA/ArA ratios were associated with a significant growth improvement. The highest proportion of polyunsaturated fatty acids was found in WB neural tissue (approx. 50% of total fatty acids), while HSB neural tissue contained the highest proportion of saturated fatty acids (approx. 35% of total fatty acids). Within the neural tissues of all Morone larvae, both DHA and ArA were generally the most dominant as well as the most responding fatty acids to dietary manipulations (except in WB fed DHA or ArA deficient diets). HSB neural tissue was particularly efficient in retaining a significant amount of DHA in the face of dietary deficiency. However, WB neural tissue was the most responsive to dietary increase in DHA, accumulating a significantly higher amount of DHA (P<0.05) than SB or HSB. Results demonstrate significant differences in fatty acid composition and growth responsiveness to dietary manipulations between Morone larvae species and within specific tissues. WB weight gain and neural tissue composition was affected most by dietary changes in both DHA and ArA whereas SB and HSB tissue compositions were generally less affected by dietary manipulations.     

Effect of salinity on expression of branchial ion transporters in striped bass (Morone saxatilis)

The time course of osmoregulatory adjustments and expressional changes of three key ion transporters in the gill were investigated in the striped bass during salinity acclimations. In three experiments, fish were transferred from fresh water (FW) to seawater (SW), from SW to FW, and from 15-ppt brackish water (BW) to either FW or SW, respectively. Each transfer induced minor deflections in serum [Na+] and muscle water content, both being corrected rapidly (24 hr). Transfer from FW to SW increased gill Na+,K+-ATPase activity and Na+,K+,2Cl- co-transporter expression after 3 days. Abundance of Na+,K+-ATPase alpha-subunit mRNA and protein was unchanged. Changes in Na+,K+,2Cl- co-transporter protein were preceded by increased mRNA expression after 24 hr. Expression of V-type H+-ATPase mRNA decreased after 3 days. Transfer from SW to FW induced no change in expression of gill Na+,K+-ATPase. However, Na+,K+,2Cl- co-transporter mRNA and protein levels decreased after 24 hr and 7 days, respectively. Expression of H+-ATPase mRNA increased in response to FW after 7 days. In BW fish transferred to FW and SW, gill Na+,K+-ATPase activity was stimulated by both challenges, suggesting both a hyper- and a hypo-osmoregulatory response of the enzyme. Acclimation of striped bass to SW occurs on a rapid time scale. This seems partly to rely on the relative high abundance of gill Na+,K+-ATPase and Na+,K+,2Cl- co-transporter in FW fish. In a separate study, we found a smaller response to SW in expression of these ion transport proteins in striped bass when compared with the less euryhaline brown trout. In both FW and SW, NEM-sensitive gill H+-ATPase activity was negligible in striped bass and approximately 10-fold higher in brown trout. This suggests that in striped bass Na+-uptake in FW may rely more on a relatively high abundance/activity of Na+,K+-ATPase compared to trout, where H+-ATPase is critical for establishing a thermodynamically favorable gradient for Na+-uptake.     

Intestinal uptake of lipovitellin from brine shrimp (Artemia franciscana) by larval inland silversides (Menidia beryllina) and striped bass (Morone saxatilis)

Intestinal uptake of lipovitellin (LV) from brine shrimp (Artemia franciscana) in larval inland silversides (Menidia beryllina) and striped bass (Morone saxatilis) was described using immunocytochemistry. Polyclonal antisera were raised against two subunits of LV (LV68 and LV190). When tested by immunocytochemistry, anti-LV68 showed cross-reactivity with some of the pancreatic cells especially in inland silversides. Therefore anti-LV190 was used to localize immunoreactive LV. Inland silversides at 14 days after hatching were fed Artemia nauplii and then sampled 4, 8, 12 hr after feeding. Similar experiments were carried out by using striped bass at 5 days and 15 days of age. They were sampled at 2, 4, 8, and 12 hr after feeding. Anterior enterocytes showed no evidence of uptake; however, the brush border of the cells of inland silversides reacted with the antiserum. Posterior enterocytes took up the LV and/or, possibly, their immunoreactive breakdown products. The pattern of uptake included accumulation in supranuclear vacuoles and digestion in supranuclear vacuoles, as suggested by the decay of the immunoreactivity over time. Thus, the posterior intestine of these larval fishes is the site of uptake and digestion of LV, an important nutritive component in the food of many larval fishes; this supports earlier findings using non-nutritive marker proteins.     

Effects of glycine and alanine on short-term storage and cryopreservation of striped bass (Morone saxatilis) spermatozoa

Three experiments were designed to examine the effects of the amino acids glycine and alanine on short-term storage and cryopreservation of striped bass spermatozoa. In the first experiment, the effect of glycine on post-equilibration motility was evaluated. In the presence of 2.5 or 5.0% Me(2)SO, glycine treatments (25, 50, and 75 mM) yielded higher (P<0.05) post-equilibration motility at all equilibration times examined compared to the control. There was no difference (P>0.05) among these three glycine treatments. In the second experiment, glycine and alanine at concentrations of 25, 50, 75, or 100mM were evaluated for post-thaw motility in the presence of 2.5 or 5% Me(2)SO. When compared to the control, both the glycine and alanine treatments showed positive effects on post-thaw motility at all concentrations tested. The highest (P<0.05) post-thaw motility was achieved with 50mM glycine or 75 mM alanine using 5% Me(2)SO. No interaction (P>0.05) between Me(2)SO and glycine or alanine was observed, indicating that the effect of glycine or alanine was independent of the concentrations of Me(2)SO. In the third experiment, glycine was evaluated for sperm motility, after short-term refrigerated storage and after cryopreservation of the same refrigerated semen. Sperm motility decreased after 24h of refrigerated storage in 50mM glycine treatment and the control, when compared to fresh sperm motility. However, 50mM glycine treatment yielded higher (P<0.01) sperm motility after both 24 and 48 h of storage as well as higher (P<0.01) post-thaw motility when compared to the control. An average of 30+/-2.9% and 16+/-2.4% post-thaw motility was achieved with the 50mM glycine treatment after 24 and 48 h of refrigerated semen, respectively.     

Effects of sublethal concentrations of zinc and benzene on striped bass, Morone saxatilis (Walbaum), infected with larval Anisakis nematodes

Young of the year striped bass, Morone saxatilis , were exposed to sublethal concentrations of zinc and benzene and infected with larval Anisakis nematodes to determine the effects of these stressors on various physiological parameters. Results indicated that both pollutants and parasites significantly decreased haematocrit values. Parasites alone increased the liver somatic index, decreased haematocrits and, in the initial phase of the experiment, lowered antibody titres. Pollutants alone did not appear to significantly affect antibody titres, nor was there any difference in zinc and benzene uptake in the livers of infected and non-infected fish.     

Allometric scaling of maximal enzyme activities in the axial musculature of striped bass, Morone saxatilis (Walbaum)

It had been suggested that the activity of anaerobic enzymes in the white muscle of fish increases exponentially with body size to meet the increasing hydrodynamic costs of burst swimming. We tested whether this relationship holds across a very large size range of striped bass, spanning a nearly 3,000-fold range in body mass. We examined the scaling of marker enzymes of anaerobic (lactate dehydrogenase and pyruvate kinase) and aerobic (citrate synthase and malate dehydrogenase) metabolism in the red and white locomotor muscles. In white muscle, we found positive scaling of anaerobic enzymes only in smaller fishes. Positive scaling of anaerobic enzymes was not found among the samples that included fishes >1,000 g despite having a sufficiently large sample size to detect such scaling. The absence of positive scaling in the white muscles of large bass suggests that they are unable to generate sufficient power to sustain relative burst swimming performance. Enzymes from aerobic pathways had activities that were mass independent in both red and white muscle. Red and white muscles were metabolically distinct except among the smallest fishes. Among young of the year, the anaerobic capacity of red muscle approached that of white muscle and also showed positive scaling. This unusual pattern suggests that red muscle might augment white muscle during burst swimming and add to the total power generated by these small fish. Maximizing burst swimming performance may be critical for small fishes vulnerable to predation but unimportant for large fishes.     

Dietary oligonucleotides from yeast RNA influence immune responses and resistance of hybrid striped bass (Morone chrysops x Morone saxatilis) to Streptococcus iniae infection

Three feeding trials were conducted to evaluate potential immunomodulatory effects of nucleotides in the diet of hybrid striped bass. A basal diet was formulated from menhaden fish meal to contain 40% crude protein and 10% lipid. An oligonucleotide product (Ascogen P) from brewer's yeast was added to the basal formulation at the manufacturer's recommended rate of 0.5% to produce the experimental diet. Each diet was fed to four replicate groups of juvenile hybrid striped bass for seven or eight weeks in two separate trials. After Trials 1 and 2, a Streptococcus iniae bath challenge was executed to test the effects of diet on disease resistance. No significant difference in growth performance was observed between fish fed the basal and experimental diets. Body composition of whole fish, hematocrit and serum lysozyme levels were observed to be within normal ranges and not influenced by dietary nucleotides. Neutrophil oxidative radical production of fish fed the nucleotide-supplemented diet was significantly (P=0.011) higher than in fish fed the basal diet. Significantly (P<0.05) enhanced survival after exposure to S. iniae also was generally observed in fish fed the nucleotide-supplemented diet. In addition, fish fed the nucleotide-supplemented diet tended to have a higher antibody response based on microtitration agglutination; however, the difference was not statistically significant because of high variation between individual fish. Long-term (16 weeks) administration of oligonucleotides in Trial 3 failed to show enhancement of immune responses between treatments. It is concluded that dietary oligonucleotides positively influenced immune responses and resistance of juvenile hybrid striped bass to S. iniae infection.     

Lesion induction by the plerocercoid Lacistorhynchus tenuis (Cestoda) and wound healing in the striped bass Morone saxatilis (Walbaum)

San Francisco Bay-Delta striped bass, Morone saxatilis (Walbaum), form open lesions in response to a plerocercoid infection of Lacistorhynchus tenuis (Van Beneden, 1858) (Cestoda: Trypanorhyncha). Laboratory infection experiments showed that striped bass can be infected with the plerocercoids by ingesting infected copepods. Histological sections indicated that a cellular host response was mounted early in the infection period, and that despite the leucocytic infiltration the parasites continued to develop. However, at 3 months post-infection some of the plerocercoids began to degenerate, and lesions formed at this time and 14 months post-infection. Open lesions in adult striped bass collected from the field took 2 months to heal and were detectable for at least 22 months. Regeneration of the muscle tissue did not occur although the wound completely healed externally.     

Effects of dimethyl sulfoxide and glycine on cryopreservation induced damage of plasma membranes and mitochondria to striped bass (Morone saxatilis) sperm

Intact plasma membrane and functional mitochondria are important attributes for the fertilization capacity of fish sperm. In the present study, dimethyl sulfoxide (Me(2)SO) and glycine were investigated in an effort to improve plasma membrane integrity and mitochondrial function in cryopreserved striped bass (Morone saxatilis) sperm. Prior to freezing, no concentration of Me(2)SO (2.5, 5 or 10%) was found to affect (P>0.05) the integrity of plasma membranes after sperm were exposed for 10 min. However, mitochondrial function decreased (P>0.05) with increasing Me(2)SO concentration. Both fluorescent staining and microscopic examination of the ultrastructure of post-thaw plasma membranes indicated that with increasing Me(2)SO concentration, plasma membranes were better protected, and 10% Me(2)SO had the highest percentage of sperm with plasma membranes intact. However, sperm mitochondrial function decreased (P>0.05) with increasing Me(2)SO concentration. The inverse relationship between plasma membrane integrity and mitochondrial function, given the Me(2)SO concentration, suggests that care must be taken to select Me(2)SO concentration that will maximize the protection of both plasma membranes and mitochondrial function. The addition of glycine to the cryomedia increased (P<0.05) the percentage of sperm with post-thaw functional mitochondria and ATP content. However glycine did not provide (P<0.05) protection to post-thaw plasma membrane integrity. The highest percentage of sperm with both intact plasma membranes and functional mitochondria was obtained with 7.5% Me(2)SO and 75 mM glycine.     

Evaluation of DNA pooling for the estimation of microsatellite allele frequencies: a case study using striped bass (Morone saxatilis)

Using striped bass (Morone saxatilis) and six multiplexed microsatellite markers, we evaluated procedures for estimating allele frequencies by pooling DNA from multiple individuals, a method suggested as cost-effective relative to individual genotyping. Using moment-based estimators, we estimated allele frequencies in experimental DNA pools and found that the three primary laboratory steps, DNA quantitation and pooling, PCR amplification, and electrophoresis, accounted for 23, 48, and 29%, respectively, of the technical variance of estimates in pools containing DNA from 2-24 individuals. Exact allele-frequency estimates could be made for pools of sizes 2-8, depending on the locus, by using an integer-valued estimator. Larger pools of size 12 and 24 tended to yield biased estimates; however, replicates of these estimates detected allele frequency differences among pools with different allelic compositions. We also derive an unbiased estimator of Hardy-Weinberg disequilibrium coefficients that uses multiple DNA pools and analyze the cost-efficiency of DNA pooling. DNA pooling yields the most potential cost savings when a large number of loci are employed using a large number of individuals, a situation becoming increasingly common as microsatellite loci are developed in increasing numbers of taxa.     

Reconciling nuclear microsatellite and mitochondrial marker estimates of population structure: breeding population structure of Chesapeake Bay striped bass (Morone saxatilis)

Comparative analyses of nuclear and organelle genetic markers may help delineate evolutionarily significant units or management units, although population differentiation estimates from multiple genomes can also conflict. Striped bass (Morone saxatilis) are long-lived, highly migratory anadromous fish recently recovered from a severe decline in population size. Previous studies with protein, nuclear DNA and mitochondrial DNA (mtDNA) markers produced discordant results, and it remains uncertain if the multiple tributaries within Chesapeake Bay constitute distinct management units. Here, 196 young-of-the-year (YOY) striped bass were sampled from Maryland's Choptank, Potomac and Nanticoke Rivers and the north end of Chesapeake Bay in 1999 and from Virginia's Mataponi and Rappahannock Rivers in 2001. A total of 10 microsatellite loci exhibited between two and 27 alleles per locus with observed heterozygosities between 0.255 and 0.893. The 10-locus estimate of R(ST) among the six tributaries was -0.0065 (95% confidence interval -0.0198 to 0.0018). All R(ST) and all but one theta estimates for pairs of populations were not significantly different from zero. Reanalysis of Chesapeake Bay striped bass mtDNA data from two previous studies estimated population differentiation between theta=-0.002 and 0.160, values generally similar to mtDNA population differentiation predicted from microsatellite R(ST) after adjusting for reduced effective population size and uniparental inheritance in organelle genomes. Based on mtDNA differentiation, breeding sex ratios or gene flow may have been slightly male biased in some years. The results reconcile conflicting past studies based on different types of genetic markers, supporting a single Chesapeake Bay management unit encompassing a panmictic striped bass breeding population.     

Ontogeny of follicle-stimulating hormone and luteinizing hormone gene expression during pubertal development in the female striped bass, Morone saxatilis (Teleostei)

Pubertal development in teleost fish is characterized by gonadal growth that is directly stimulated by the pituitary gonadotropins, FSH and LH. We used a quantitative ribonuclease protection assay to provide, for the first time, the developmental profiles of the alpha-, betaFSH-, and betaLH-subunit gene expression in a seasonal breeding fish, the female striped bass (3-yr study, n = 207). Two-year-old females were sexually immature, although a transient rise in all gonadotropin subunit mRNAs was measured in the pituitary. Pubertal ovarian development occurred in 65% of 3-yr-old females, characterized by the appearance of lipid droplets within the oocytes. This reproductive phase, termed pubertal development, was associated with a 34-fold increase in the mRNA levels of betaFSH and a rise in the pituitary concentration of LH. The first sexual maturation took place in 4-yr-old females and coincided with a 218-fold increase in the mRNA levels of betaFSH. During this time period, the mRNA levels of the alpha and betaLH subunits increased by 11- and 8-fold, respectively. At the final stages of vitellogenic growth, mRNA levels of betaFSH declined to basal levels, whereas the mRNA levels of the alpha and betaLH subunits remained elevated. Throughout the study, pituitary LH concentration was positively correlated to the mRNA levels of betaLH, but plasma levels of LH remained low and unchanged (0.4-0.8 ng/ml) despite increasing levels of pituitary LH concentration, suggesting a regulated secretion pathway. Taken together, the data show that the profiles of betaFSH and betaLH mRNAs appear to follow an annual rhythm that is associated with developmental events in the growing oocytes. In particular, increasing levels of betaFSH mRNA appear to underlie the first sexual maturity in the female striped bass.