Population genetics of the American oyster Crassostrea virginica along the Atlantic coast and the Gulf of Mexico

An examination by protein-gel electrophoresis of 19 different geographical populations of the American oyster Crassostrea virginica (Gmelin) was conducted along the Atlantic coast and the Gulf of Mexico. Estimates were made of levels of genetic variation and similarity among the populations based on 32 structural loci. The percentage of loci polymorphic ranged from 46.9 to 65.6% along the Atlantic coast while the estimate ranged from 54.8% to 68.8% on the Gulf of Mexico. The percentage of loci heterozygous ranged from 18.6 to 23.6% along the Atlantic coast and from 20.0 to 25.4% in the Gulf of Mexico. The genetic similarities between all contiguous populations from Cape Cod, Massachusetts to Corpus Christi, Texas were estimated as 99%, while the same estimate between Corpus Christi and Brownsville, Texas was computed as 93%, indicating a major transition in genetic structure for the Brownsville population of the Laguna Madre. The study revealed that the migration of planktonic oyster larvae is predominantly in a westerly direction along the Gulf of Mexico and that gene flow appears to be disrupted in the region of the Laguna Madre. Evidence has been presented for single gene selection at the Lap-2 and Pgi loci in the form of macrogeographical clines in allele frequencies with changing environmental conditions.     

Sequence and atomic-force microscopy analysis of a matrix protein from the shell of the oyster Crassostrea virginica

Atomic-force microscopy of the major class of soluble matrix protein from the oyster Crassostrea virginica revealed that this protein forms a ring structure, perhaps rendering the NH₂-terminus unavailable to Edman degradation. Cleavage of these proteins using mild acid hydrolysis and hydroxylamine produced linear peptides that were able to be sequenced. Peptides consisted of a domain of polyserine-phosphoserine and runs of aspartic acid of 4 to 7 residues, with several other amino acids present. Hydrophobic domains were also isolated. Although phylogenetically distant, oyster shell-matrix protein is similar to the phosphophoryn isolated from dentin and, to a lesser extent, proteins isolated from verteorate bone.     

Dietary protein levels and growth of the oyster Crassostrea virginica

The diatom Thalassiosira pseudonana in exponential phase or stationary phase and grown under 3 light regimes was fed to 6 groups of juvenile oysters, Crassostrea virginica Gmelin, for a 10-week period. Oysters fed diets from the stationary phase grew more rapidly and had a higher glycogen content than those fed diets from the exponential phase. The stationary phase, cool white-light diet produced the greatest increase in dry weight, shell height and glycogen content of the 6 diets examined. Oysters grew more rapidly when fed diets richer in carbohydrates than proteins.     

Accumulation and depuration of mercury in the American oyster Crassostrea virginica

To study the kinetics of mercury uptake in oysters, adult Crassostrea virginica (Gmelin) were held in seawater containing 10 g mercury/l (ppb) or 100 g mercury/l (ppb), added in the form of mercuric acetate, for 60 days. Mercury concentration in tissues was determined by analysis of individually homogenized oyster meats, using wet digestion and flameless absorption spectrophotometry. After 45 days, average mercury tissue concentration was 140,000 g mercury/kg tissue (ppb) and 28,000 g mercury/kg tissue (ppb) in the 100 ppb and 10 ppb experimental groups, respectively. After this time, concentrations dropped sharply, probably due to spawning. Clearance of mercury from tissue was studied by exposing treated adults to estuarine water (with no additions) for 30 days (100 ppb group) and 160 days (10 ppb group). Tissue concentrations in the 100 ppb mercury environment group declined from 115,000 to 65,000 ppb, and those of the 10 ppb group declined from 18,000 to 15,000 ppb, in 18 days; there-after, no further decline occurred in either group. Oysters accumulated mercury 1,400 times and 2,800 times above the environmental concentrations of 100 and 10 ppb mercury, respectively. Total self-purification was not achieved over a 6 month cleansing period.     

Ultrastructure of the gonad and gametogenesis in the eastern oyster,Crassostrea virginica. I. Ovary and oogenesis

The ultrastructural features of the ovary and oogenesis are described in the eastern oyster,Crassostrea virginica (Gmelin, 1791). The ovary is a diffuse organ consisting of highly branching acini in which oocytes develop. The acini are surrounded by a matrix of vesicular connective tissue (VCT tells) which serves a nutrient storage function. Each acinus is bathed by fluid within a surrounding connective tissue compartment, the hemocoel, which likely serves as a means of transporting nutrients to the oocytes. Oocytes begin growth while positioned near to the inner acinus wall. As differentiation proceeds and they enter the late stages of vitellogenesis, they become stalle-shaped and project into the acinus lumen. Follicle tells are closely associated with oocytes during the early and middle stages of vitellogenesis but they are largely confined to the basal, stalked region of late-stage oocytes. Vitellogenesis occurs through a process of autosynthesis, involving the combined activity of the Golgi complex and rough endoplasmic reticulum, and heterosynthesis in which extraovarian precursors are incorporated into oocytes via receptor-mediated endocytosis involving the basal surface of the oocytes. It is suggested that the follicle tells play some important role during oogenesis but probably are not the major source of yolk precursors. The VCT celas are probably the main source of nutrients for vitellogenesis.     

Ultrastructure of the gonad and gametogenesis in the eastern oyster,Crassostrea virginica. II. Testis and spermatogenesis

The pelagic harpacticoid copepod, Macrosetella gracilis (A. Scott), is found in association with colonies of the nitrogen-fixing (diazotrophic), bloomforming cyanobacterium Trichodesmium spp. in tropical and subtropical waters. M. gracilis is one of the few direct grazers of these often toxic cyanobacteria. Experiments investigating NH ₊ ⁴ regeneration by M. gracilis were conducted in the Caribbean in September 1992 and the Coral Sea, Australia in November 1994. Rates of M. gracilis ingestion of Trichodesmium thiebautii labelled with ¹⁵N₂ measured in the eastern Caribbean indicated that M. gracilis could consume 33 to 45% of total T. thiebautii colony N d^-1 and >100% of new N fixed d^-1. We also measured the release of NH ₊ ⁴ by M. gracilis feeding on T. thiebautii, as well as by non-feeding copepods, using ¹⁵N isotope dilution methods. In non-feeding copepods, rates of NH ₊ ⁴ release increased as numbers of copepods were increased as both copepod numbers and food availability increased. In the presence of T. thiebautii colonies, M. gracilis had an average rate of NH ₊ ⁴ regeneration of 7.7±1.5 nmol N copepod^-1 h^-1 (±SE), which was significantly higher than when food was absent (1.9±0.7 nmol N copepod^-1 h^-1). Rates of M. gracilis excretion were relatively high based on excretion: ingestion ratios, which could be due to having a high-N food source readily available, to sloppy-feeding effects, or as a response to toxins in the cyanobacterium. Incubations of M. gracilis with and without T. erythraeum resulted in significant increases in [NH ₊ ⁴ ] as a function of copepod density only. Ammonium leakage from the cyanobacterium and/or microheterotroph associates was relatively low. M. gracilis, through excretion and possible mechanical breakage of cells while grazing, appears to provide a direct link between atmospherically derived new nitrogen and regenerated NH ₊ ⁴ in the oligotrophic systems where Trichodesmium spp. are abundant.     

Interaction of genotype and salinity in larvae of the oyster Crassostrea virginica

Adult Crassostrea virginica were obtained from 4 populations and spawned in the laboratory. The larvae from the within-population crosses and the hybrid crosses were raised at 4 salinities. There were no significant differences in survival of the larvae between the populations. However, one set of hybrids did show overdominance in survival. There were genetic differences between the populations in growth rate, but the expression of the differences depended upon the salinity, i.e, there was significant genotype-environment interaction. There were expressions of nonadditive genetic effects in the hybrid crosses, but the direction and magnitude was dependent upon the salinity. There was as much difference between populations from the same estuary as there was between populations from geographically isolated populations.     

Genetics of larvae and spat growth rate in the oyster Crassostrea virginica

The heritability of oyster (Crassostrea virginica) larval growth rate was estimated to be in the range of 0.25 to 0.50 and a significant part of this genetic variation is of the additive type. Larval growth rate and spat growth rate were found to be highly correlated. These results suggest that a selection program for faster growing larvae and spat would be successful.     

Inducible defenses in the eastern oyster Crassostrea virginica Gmelin in response to the presence of the predatory oyster drill Urosalpinx cinerea Say in Long Island Sound

The response of the eastern oyster C. virginica to the presence of the oyster drill Urosalpinx cinerea was examined from July to September 2011. Several aspects of oyster growth were measured, including wet weight, shell weight, and dorsal shell area for oysters collected near Groton, Connecticut (41.32036 N, −72.06330 W). Wet weight and shell weight growth were significantly higher in the presence of the predator U. cinerea, while tissue weight showed no difference from the control. The control group showed more shell area growth and a much lower ratio of shell weight growth to shell area growth. Differences in shell weight to area ratio indicated that C. virginica dramatically shifted from lateral shell growth to shell thickening in the presence of U. cinerea. This inducible defense has not been previously shown for C. virginica and could play an important role in the predator–prey interaction between these two species.     

Accumulation,release and retention of petroleum hydrocarbons by the oyster Crassostrea virginica

Two Crassostrea virginica populations, differing in fat content, were experimentally exposed to a complex petroleum-hydrocarbon fraction. The hydrocarbons in this mixture were accumulated by both groups of oysters, and their lipid content, as well as the concentration of hydrocarbon in the water, were found to affect the rate and extent of accumulation. Hydrocarbons accumulated were rapidly, although incompletely, discharged when the oysters were transferred to an uncontaminated system. Amounts of hydrocarbons discharged and amounts retained after discharge are probably related to the level of contamination. The data can be interpreted as indicating that equilibration and the occurrence of multiple compartments where hydrocarbons can reside are factors involved in the uptake and retention of nonbiogenic hydrocarbons by oysters. The petroleum hydrocarbons contained in the oysters differed from the contaminating oil by displaying a greater aromatic content. In addition, gas-liquid chromatograms of aliphatic fractions of the hydrocarbons in the oysters rapidly showed a degraded appearance; the possibility that the oysters themselves are modifying the oil cannot be excluded.Contribution No. 3098 from the Woods Hole Oceanographic Institution.     

Effects of the dinoflagellates Karlodinium veneficum and Prorocentrum minimum on early life history stages of the eastern oyster (Crassostrea virginica)

The bloom-forming dinoflagellates Prorocentrum minimum and Karlodinium veneficum can have detrimental effects on some marine life, including shellfish, but little is known about their effects on early life history stages of bivalves. In the Chesapeake Bay region, blooms of these dinoflagellates overlap with the spawning season of the eastern oyster, Crassostrea virginica. In laboratory experiments, we compared the effects of P. minimum and K. veneficum on the survival and development of embryos and larvae of the eastern oyster. At 10⁴ cells ml⁻¹, P. minimum did not have a negative effect on embryos and larvae in 2-day exposures. The yield of D-hinge larvae was equal to or greater than in control treatments. At 2 × 10⁴ cells ml⁻¹ (approximately equal biomass to the P. minimum treatment) K. veneficum caused significant mortality to oyster embryos within 1 day and almost no embryos developed into D-hinge larvae. This effect was not alleviated by the provision of an alternate food source (Isochrysis sp.). Significant mortality was observed when larvae were exposed to K. veneficum at concentrations of 10⁴ cells ml⁻¹ (approximately 5 ng ml⁻¹ of karlotoxin). The K. veneficum cultures used in these experiments were relatively low in toxin content, more toxic strains could be expected to cause mortality at lower cell concentrations. Survival and maturation of embryos and larvae may be reduced when spawns of the eastern oyster coincide with high bloom densities of K. veneficum.     

The toxicity of heavy metals to embryos of the American oyster Crassostrea virginica

The acute toxicity of 11 heavy metals to embryos of the American oyster Crassostrea virginica was studied and the concentrations at which 50% of the embryos did not develop were determined. The most toxic metals and their LC₅₀ values were mercury (0.0056 ppm), silver (0.0058 ppm), copper (0.103 ppm) and zinc (0.31 ppm). Those metals that were not as toxic and their LC₅₀ values were nickel (1.18 ppm), lead (2.45 ppm) and cadmium (3.80 ppm). Those metals that were relatively non-toxic and their LC₅₀ values were arsenic (7.5 ppm), chromium (10.3 ppm) and manganese (16.0 ppm). Aluminum was non-toxic at 7.5 ppm, the highest concentration tested.     

Uptake of Kepone from sediment suspensions and subsequent loss by the oyster Crassostrea virginica

Marine Biology - Oysters in laboratory trays received sediment suspensions prepared with sediments contaminated with Kepone** from the James River, Virginia, USA. Oysters in trays were also exposed...     

Effects of the polychlorinated biphenyl Aroclor® 1254 on the American oyster Crassostrea virginica

Young oysters (Crassostrea virginica) were continuously exposed to Aroclor^® 1254, a polychlorinated biphenyl (PCB), in flowing, unfiltered seawater. Growth rate (height and in-water weight) was significantly reduced (=0.05) in oysters exposed to 5 g/l (ppb) for 24 weeks. Growth rate was not affected in oysters exposed to 1 ppb for 30 weeks. Mortality was not significant in exposed and control groups. In oysters exposed to 5 ppb, greatest PCB residue (whole body) was 425 mg/kg (ppm), 85,000x the concentration in the water, and less than 0.3 ppm was retained after 28 weeks depuration in PCB-free water. In oysters exposed to 1 ppb, greatest residue was 101 ppm, 101,000x the concentration in the water, and less than 0.2 ppm was retained after 12 weeks depuration. Examination of oysters exposed to 5 ppb of this PCB for pathogenesis revealed atrophy of digestive diverticular epithelium and degeneration of vesicular connective tissues concomitant with leukocytic infiltration, but tissue recovery seemed excellent after 12 weeks depuration.Registered trademark, Monsanto Company, St. Louis, Missouri, USA. Mention of commercial products or trade names does not constitute endorsement by the Environmental Protection Agency.Contribution No. 146, Gulf Breeze Laboratory.     

Importance of refractory plant material to the carbon budget of the oyster Crassostrea virginica

The ability of the oyster Crassostrea virginica (Gmelin) to filter, ingest and assimilate ¹⁴C-labeled Spartina alterniflora as a carbon source was investigated under laboratory conditions. The oyster assimilated crude-fiber carbon extracted from S. alterniflora with an efficiency of approximately 3%. Enteric bacteria did not enhance this process. The annual average (April 1984 to November 1985) of crude fiber in the Choptank River sub-estuary of the Chesapeake Bay, Maryland, USA, from which the oysters were collected, was 15.7 g l^-1 (range 4.3 to 34.3 g l^-1). The potential food value of crude fiber to oysters in this system was estimated to be less than 1% of their carbon demand. However, the potential contribution of crude fiber to the carbon requirements of other oyster populations, such as those in southeastern USA, may be as great as 20%, due to higher crudefiber concentrations in the seston.     

The ecological energies of growth,respiration and assimilation in the intertidal American oyster Crassostrea virginica

Seasonal variations in the growth, respiration and assimilation of the intertidal oyster Crassostrea virginica (Gmelin) of different sizes were determined. The instantaneous growth rates for intertidal oysters decreased with increasing size and with lower temperatures. Q₁₀ values computed from instantaneous growth rates were approximately 2 during the warm growing season, but were higher in the colder months. Oxygen consumption increased with temperature and body size. A model was developed to predict oxygen consumption at any environmental temperature from 10° to 30°C for oysters ranging in weight from 0.1 to 100.0 g. Q₁₀ values computed from oxygen-consumption rates decreased with increasing temperature and increasing body size. Intertidal oysters utilize a large proportion of their assimilated energy in growth.Supported by a Belle W. Baruch Fellowship in Marine Ecology.     

Factors affecting the accumulation and removal of mercury from tissues of the American oyster Crassostrea virginica

Adult oysters, Crassostrea virginica (Gmelin) were held in seawater containing 10 or 100 ppb mercury in the form of mercuric acetate for 45 days. Mercury concentration in tissues was determined by analysis of individually homogenized oyster meats using wet digestion and flameless absorption spectrophotometry. After 45 days, average mercury tissue concentration was 91,600 and 12,100 ppb in the 100 and 10 ppb mercury groups, respectively. A slight decline in mercury residues in the 100 ppb group during the accumulation period was attributed to spawning. Clearance of mercury from tissues was studied in a constant temperature regime (25°C±2C^o) for 25 days and in a declining temperature regime (25° to 5°C) for 80 days by exposing treated adults to estuarine water with no mercury added. The biological half-life of mercuric acetate was 16.8 and 9.3 days in the 25°C temperature regime, and 35.4 and 19.9 days in the declining temperature regime, for the 10 and 100 ppb groups, respectively. Smaller oysters (0 to 7 g) consistently accumulated more mercury per gram wet weight than larger oysters (7 to 20 g) in populations exposed to 10 and 100 ppb mercury.     

Winter quiescence and spring awakening of the Eastern oyster Crassostrea virginica at its northernmost distribution limit

To test the hypothesis that oysters, Crassostrea virginica, from the northernmost part of the species range in the Gulf of St. Lawrence (48°N) open their valves at lower temperatures than those reported for more southern oysters, Hall element sensors were used to monitor their gaping behaviour. These observations were made in a flow-through system and the temperature, salinity and relative fluorescence of unfiltered seawater were monitored. Photoperiod was controlled (15?h dark:9?h light) and light levels were measured but not closely controlled. Gaping behaviour was followed from February to June 2010 (113?days) and from April to May 2011 (34?days) and was classified into three successive phases: quiescent, awakening and active. Although valves were either closed or slightly open during the quiescent phase (maximum gape angle?=?0.49°, SE?=?0.04), they abruptly opened to maximum angles of about 5.88° (SE?=?0.29) during the awakening phase. Moreover, there was noticeable synchrony amongst individuals, since approximately one-half of the monitored population awoke within a 6.6-h period in both study years. Correlative analyses identified temperature as a factor influencing valve movement, and oysters awakened when temperatures were 0.2–4.0?°C (mean?=?2.2, SE?=?0.2). Oysters exerted their maximal gape angle as soon as temperatures reached 2.8–6.6?°C (mean?=?4.8, SE?=?0.2). During the active phase, valves remained open 68.6?% (2010) and 79.7?% (2011) of the time. An unexpected result was the observation of a diurnal rhythm in valve openness whereby the openness was greatest near the end of the afternoon and least in the early morning.     

Rate of excretion of ammonia by the hard clam Mercenaria mercenaria and the American oyster Crassostrea virginica

Two species of bivalve molluscs, Mercenaria mercenaria Linné and Crassostrea virginica Gmelin, were maintained in an identical laboratory environment and fed the same diet for 1 month prior to measuring the quantity of ammonia which they excreted per gram dry weight. Data derived from juveniles and adults of these species were fitted to a log-log equation. M. mercenaria excreted more ammonia per gram body weight than C. virginica, and considerably more scatter is evident in the fit of the data from M. mercenaria to the log-log relationship. Experiments showed that neither temperature fluctuation during the experiments nor decomposition of organic nitrogen in the test water account for the scatter of points. Behavioral differences between the two species of organisms may explain these differences.This work is the result of research sponsored by NOAA Office of Sea Grant, Department of Commerce, under Grant No. 04-3-158-30.     

Effects of tidal fluctuations of salinity on pericardial fluid composition of the American oyster Crassostrea virginica

Crassostrea virginica Gmelin were subjected to simulated tidal fluctuations of salinity, and the subsequent effects on osmotic and ionic composition of the pericardial fluid, body water and valve movements were investigated. Ambient salinity fluctuation patterns of 20-10-20, 15-10-15 and 10-5-10 were simulated during 24.8-h periods. An additional 10-5-10 S experiment was performed using a dilution water approximating the ionic composition of Mississippi River water with regard to Mg⁺⁺, Ca⁺⁺ and SO ₄ ⁼ , instead of deionized water. Finally the effects of a 2-week diurnal fluctuation pattern between 20 and 10 S were investigated with respect to pericardial fluid composition. Pericardial fluid osmolality, concentrations of Cl^-, Na⁺, Mg⁺⁺, K⁺, Ca⁺⁺ and ninhydrin-positive substances (NPS) were analyzed periodically throughout all experiments. Pericardial fluid osmolality was slightly hyperosmotic as ambient water salinity decreased during a cycle, and then became slightly hyposmotic as ambient salinity increased. In the 2-week experiment, pericardial fluid osmolality tracked ambient seawater closely through Day 5, but became more intermediate between high and low seawater values as the experiment progressed. Similar patterns during fluctuations of salinity were observed for Na⁺, Cl^-, Mg⁺⁺ and Ca⁺⁺. Pericardial fluid K⁺ levels did not track ambient seawater as closely as did other ions. The ionic composition of dilution water had little effect on the osmotic or ionic response of the oyster's pericardial fluid. Pericardial fluid NPS level varied inversely with salinity during the 20-10-20 cycle. During the longterm fluctuation experiment, NPS values gradually decreased over the 2-week period compared to constant salinity control values. Percent body water also varied inversely with ambient salinity. Solute movement accounted for most of the change in pericardial fluid osmolality during the simulated cycles with water movement responsible for 1 to 11%. Water movement contributed more to the change of pericardial fluid osmolality during the decreasing salinity phase than the increasing phase of a given cycle. During 20-10-20 S cycles, oyster valves remained open 56% of the time (n=23). In contrast, when salinity was abruptly changed from 20 to 10 within 5 min, valve closure occurred in 4.8±0.3 min (n=20). Valves did not reopen for 19.3±1.2 h (n=15).