Metal concentration changes in growing Pacific oysters,Crassostrea gigas,cultivated in Tasmania,Australia

Pacific oyster spat (Crassostrea gigas Thunberg) transferred from the Tamar Estuary to two growing areas in southern Tasmania were monitored for their metal contents over one growing season (1974–1975). Oysters at Pipeclay Lagoon were grown with stick and tray culture while those at Dart Island were cultured with the longline technique. Metal content of the oysters increased with time and the trend was similar to the weight growth curves. Mean dry weights of oysters increased from 0.07 to 1.19 g at Pipeclay Lagoon and from 0.25 to 1.47 g at Dart Island. Metal contents (g) increased at each site, respectively: Fe 57 to 326; 91 to 446; Zn 259 to 6 555; 755 to 5 335; Cd 1.5 to 13.3; 1.9 to 16.3; Cu 26 to 142; 9 to 116; Pb 1.9 to 11.9; 0.6 to 3.8. Concentration curves generally showed a downward trend with time. The relationships of metal concentrations with weight did not differ from sample to sample at a site nor did they differ at one site compared with the other. The only exception was lead, which showed no relationship of concentration with weight at Pipeclay Lagoon and a negative one at Dart Island. It is postulated that higher winter concentrations of metals in the oysters were linked with greater solubility of metal ions in lower salinity water.     

Gametic incompatibility and genetic divergence of Pacific and Kumamoto oysters,Crassostrea gigas and C. sikamea

The biological and taxonomic separation of the Pacific oyster Crassostrea gigas (Thunberg, 1793) from the Kumamoto oyster C. sikamea (Amemiya, 1928) is affirmed by three concordant lines of evidence: (1) fixed differences in 2% of a mtDNA sequence coding for large subunit rRNA; (2) a genetic distance of 0.440 based on 19 allozyme loci, including 5 diagnostic loci (Aat-1, Idh-1, Idh-2, Mpi-1, Mdh-2); (3) one-way gametic incompatibility resulting in partial reproductive isolation in interspecific crosses. C. gigas sperm x C. sikamea egg fertilizations form viable hybrid offspring, but C. sikamea sperm do not fertilize C. gigas eggs. Divergence between these two species is mediated by differing peaks in the periods for gamete release and by one-way sperm/egg incompatibility. Two attempts to recover C. sikamea from its place of origin in southern Japan have yielded only individuals with the mitochondrial haplotype that characterizes C. gigas. We thus identify a crucial need for careful screening, management, and conservation of the cultivated populations of C. sikamea on the US west coast.     

Cadmium kinetics in oysters — a comparative study of Crassostrea gigas and Ostrea edulis

The accumulation of cadmium was investigated in two species of oysters [Crassostrea gigas (L.) and Ostrea edulis (L.)] from the same environment and in oysters of the same species (O. edulis) from two different environments (contaminated and uncontaminated), under controlled laboratory conditions (33‰ salinity, 10°C, 100 μg Cd l^-1) for up to 111 d in 1982. C. gigas accumulated cadmium twice as fast as O. edulis (1.07 vs 0.52 μg Cd g^-1 wet wt d^-1). Furthermore, O. edulis from an uncontaminated environment accumulated cadmium faster than O. edulis from a metal-contaminated environment (0.52 vs 0.34 μg Cd g^-1 wet wt d^-1). There was no effect of cadmium exposure on total soft-tissue copper and zinc concentrations. Investigation of cytosolic metal-binding using Sephadex G-75 gel-permeation chromatography indicated that binding to very low molecular weight ligands (MW<1000) accounted for>70% of the cytosolic zinc in all oysters and>40% of the cytosolic cadmium in all oysters except O. edulis from Conwy at 83 d. In copper-contaminated oysters, excess copper was also associated with very low molecular weight ligands. Intermediate molecular weight cadmium/copper-binding proteins (similar to metallothionein in molecular weight) were observed in the cytosol and were shown to differ between species in terms of their behavior on Sephadex G-75. Finally, the distribution of accumulated cytosolic cadmium in O. edulis from the contaminated environment was shown to have a unique distribution, i.e., there was no cadmium associated with high molecular weight cytosolic macromolecules. The data indicate that both genetic and environmental factors influence cadmium accumulation in oysters.     

Competence and delay of metamorphosis in the Pacific oysterCrassostrea gigas

The temporal relationship between the onset of behavioral and morphogenetic competence was determined inCrassostrea gigas (Thunberg) larvae by exposure to appropriate chemical inducers during development. Larvae exhibited settlement behavior in response to L-3,4-dihydroxyphenylalanine (L-DOPA) before becoming competent to metamorphose in response to epinephrine. For larvae past the onset of competence, the apparent stimulus-threshold decreased and they were increasingly likely to metamorphose subsequent to induced settlement behavior. In the absence of chemical stimulation, cultured oyster larvae were able to delay metamorphosisand maintain competence for at least 30 d. Competence was correlated with, but not dependent upon, larval size and eyespot development. A mechanistic model of oyster settlement and metamorphosis is proposed which incorporates these new data.Contribution # 136 from the Center of Biotechnology, Marine Biotechnology Institute, University of Maryland     

Elevated CO2 alters larval proteome and its phosphorylation status in the commercial oyster, Crassostrea hongkongensis

Ocean acidification (OA) is beginning to have noticeable negative impact on calcification rate, shell structure and physiological energy budgeting of several marine organisms; these alter the growth of many economically important shellfish including oysters. Early life stages of oysters may be particularly vulnerable to OA-driven low pH conditions because their shell is made up of the highly soluble form of calcium carbonate (CaCO₃) mineral, aragonite. Our long-term CO₂ perturbation experiment showed that larval shell growth rate of the oyster species Crassostrea hongkongensis was significantly reduced at pH < 7.9 compared to the control (8.2). To gain new insights into the underlying mechanisms of low-pH-induced delays in larval growth, we have examined the effect of pH on the protein expression pattern, including protein phosphorylation status at the pediveliger larval stage. Using two-dimensional electrophoresis and mass spectrometry, we demonstrated that the larval proteome was significantly altered by the two low pH treatments (7.9 and 7.6) compared to the control pH (8.2). Generally, the number of expressed proteins and their phosphorylation level decreased with low pH. Proteins involved in larval energy metabolism and calcification appeared to be down-regulated in response to low pH, whereas cell motility and production of cytoskeletal proteins were increased. This study on larval growth coupled with proteome change is the first step toward the search for novel Protein Expression Signatures indicative of low pH, which may help in understanding the mechanisms involved in low pH tolerance.     

Depuration of twelve trace metals in tissues of the oysters Crassostrea gigas and C. virginica

The depuration of 12 trace metals in the mantle, gill, digestive gland, and kidney of Crassostrea gigas and C. virginica was investigated under natural field conditions; oysters from a relatively contaminated environment (Redwood Creek in south San Francisco Bay) were transplanted to a relatively clean environment (Tomales Bay). In the transplanted oysters, the digestive gland and kidney depurated Cd, Cu, Hg, Ag, and Zn more readily than the mantle and gill. Other trace metals As, Fe, Mn, Ni and Se showed varying depuration patterns. The results for Cr and Pb were inconclusive, since initial concentrations were too low to follow any losses. Interspecific differences in trace metal depuration were observed. Biological half-lives for most trace metals were on the order of 23 to 60 d for C. gigas and on the order of 70 to 180 d for C. virginica.     

Preparation and evaluation of protein microcapsules for a marine suspension-feeder,the Pacific oyster Crassostrea gigas

Little is known of the dietary requirements of marine suspension-feeders, mainly due to difficulties in preparing and delivering microparticulate diets to animals living in an aquatic environment. The food particles must be stable in seawater and digestible by the consumer. A laboratory method for preparing cross-linked, protein-walled capsules is described in this study as a means of delivering dietary protein to marine suspension-feeders such as the oyster Crassostrea gigas. Leakage of ¹⁴C-labelled protein from capsules suspended in seawater for 24 h was reduced to less than 5% by the use of the trifunctional cross-linking agent 1,3,5-benzenetricarbonyl trichloride in wall formation. Invitro experiments indicated that capsules were susceptible to digestion by proteases and by style extracts of oysters. Oysters fed encapsulated ¹⁴C-labelled protein absorbed ¹⁴C with an efficiency of 40.2±5.0% (±1 SE). Biochemical fractionation of capsule-fed oysters indicated breakdown of ingested ¹⁴C-protein into material that was soluble in 7% trichloroacetic acid. Cross-linked, protein-walled capsules can be used successfully to deliver dietary protein to the oyster C. gigas.     

Gill development, functional and evolutionary implications in the Pacific oyster Crassostrea gigas (Bivalvia: Ostreidae)

Development of the Crassostrea gigas gill was studied in order to better understand the feeding biology of early life stages, identify potentially critical developmental stages which may influence rearing success or recruitment to wild populations, and shed light on the evolution of the basic bivalve gill types. Larvae and juveniles were reared in an experimental hatchery, and larger specimens were obtained from a commercial hatchery. Specimens were relaxed, fixed, dried, and observed using scanning electron microscopy (SEM). The right and left gills developed symmetrically, via a “cavitation–extension” process from the gill buds. The inner demibranchs developed first (V-stage, 0.29–2.70 mm), in a sequential postero-anterior series of homorhabdic filaments. The outer demibranchs developed later (W-stage, from 2.70 mm), also as homorhabdic filaments, synchronously along the gill axis. The principal filaments (PF) developed from the progressive fusion of three ordinary filaments (OF), at a size of 7.50 mm, and the consequent plication was accentuated by the formation of extensive tissue junctions. Effective filament number (number of descending and ascending filaments) showed a marked discontinuity at the transition from the V- to the W- stage of the gill. Filament ciliation showed several important changes: establishment of OF ciliation in the homorhabdic condition (2.70 mm), ciliary de-differentiation of the PF in the heterorhabdic condition (7.50 mm), and establishment of a latero-frontal cirri length gradient from the plical crest to the PF base. Reversal of direction of ciliary beat is also necessary prior to adult functioning of the PF. Three major transitions were identified in C. gigas gill development, each potentially important in rearing success or wild population recruitment: (1) transition from velum to gill at settlement, (2) transition from a V- to a W-shaped gill (2.70 mm), and (3) transition from the homorhabdic to the heterorhabdic condition (7.50 mm). Complete gill development was much more prolonged than in species previously studied. The major ontogenetic differences between the C. gigas heterorhabdic pseudolamellibranch gill and the pectinid heterorhabdic filibranch gill suggest that the heterorhabdic condition evolved independently in these two bivalve families.     

Comparative vulnerability to predators,and induced defense responses,of eastern oysters Crassostrea virginica and non-native Crassostrea ariakensis oysters in Chesapeake Bay

Management agencies are considering introducing the Suminoe oyster Crassostrea ariakensis into Chesapeake Bay, USA. It is unknown if the growth of feral populations of this non-native oyster would be regulated by the same predators that once controlled the abundance of the native eastern oyster C. virginica. In laboratory studies, we compared the relative susceptibility of juvenile diploids (shell height < 25 mm) of both oyster species to invertebrate predators of eastern oyster juveniles. Predators included four species of mud crabs [Rhithropanopeus harrisii (carapace width 7–11 mm), Eurypanopeus depressus (6–21 mm), Dyspanopeus sayi (8–20 mm), and Panopeus herbstii (9–29 mm)], the blue crab Callinectes sapidus (35–65 mm), and two sizes of polyclad flatworms (Stylochus ellipticus and possibly Euplana gracilis; planar area ≲5 mm² and ∼14 to 88 mm²). All four species of mud crab and the blue crab preyed significantly (ANOVA, P ≤ 0.05) more on C. ariakensis than on C. virginica, but predation by flatworms of both sizes did not differ significantly between oyster species. The greater susceptibility of C. ariakensis to crab predation was likely due to its shell compression strength being 64% lower than that of C. virginica (P = 0.005). To test for predator-induced enhancement of shell strength, we held oysters of both species for 54 days in the presence of, but protected from, C. sapidus and R. harrisii. Crabs were fed congeneric oysters twice weekly within each aquarium. Compared to controls, shell strength of C. virginica exposed to R. harrisii increased significantly (P < 0.043), as did shell strength of both oyster species exposed to C. sapidus (P < 0.01). Despite the changes in shell strength by both oyster species in the presence of C. sapidus, the shell of C. ariakensis remained 57% weaker than C. virginica. We conclude that, because C. ariakensis exposed to predators continued to have a weaker shell relative to C. virginica, the natural suite of crab and flatworm predators in Chesapeake Bay will likely serve to control the abundance of feral C. ariakensis. We caution that the situation in the natural environment may be sufficiently different in some locations that C. ariakensis may be able to compensate for its greater vulnerability to crab predation and hence become a nuisance species.     

Growth patterns in oysters, Crassostrea virginica, from different estuaries

We analyzed a data set collected over 15 yr, containing growth data from strains of eastern oysters, Crassostrea virginica (Gmelin, 1791), initiated from parent populations in Long Island Sound, Delaware Bay, and lower Chesapeake Bay. The long-term growth data proved to be a powerful tool for examining patterns of growth differentiation among separated populations of C. virginica. The oyster strains had been grown in a common environment in lower Delaware Bay for up to seven generations. We found that the oyster strains with origins in Long Island Sound were significantly larger over several generations than oyster strains from Delaware Bay and Chesapeake Bay. Chesapeake Bay oyster strains were larger than Delaware Bay oyster strains at 1.5 yr old, but Delaware Bay oysters were larger at 2.5 yr. Year-to-year variation in environmental conditions had a strong significant effect on absolute oyster size and the relative sizes of the oyster strains. Persistent differences between oyster strains from different origins over several generations support a hypothesis that these estuarine populations have experienced long-term genetically-based population differentiation. This result is consistent with hypotheses of population differentiation of oysters based on observations of local reproductive timing. Received: 12 August 1997 / Accepted: 26 May 1998     

Comparing the effect of elevated pCO2 and temperature on the fertilization and early development of two species of oysters

This study compared the synergistic effects of elevated pCO₂ and temperature on the early life history stages of two ecologically and economically important oysters: the Sydney rock oyster, Saccostrea glomerata and the Pacific oyster, Crassostrea gigas. Gametes, embryos, larvae and spat were exposed to four pCO₂ (375, 600, 750, 1,000 μatm) and four temperature (18, 22, 26, 30°C) levels. At elevated pCO₂ and suboptimal temperatures, there was a reduction in the fertilization success of gametes, a reduction in the development of embryos and size of larvae and spat and an increase in abnormal morphology of larvae. These effects varied between species and fertilization treatments with S. glomerata having greater sensitivity than C. gigas. In the absence of adaptation, C. gigas may become the more dominant species along the south-eastern coast of Australia, recruiting into estuaries currently dominated by the native S. glomerata.     

Metabolism of palmitic,linoleic, and linolenic acids in adult oysters,Crassostrea virginica

This study investigated incorporation and metabolism of saturated [(1-¹⁴C) 16:0] and unsaturated [(1-¹⁴C) 18:26 and (1-¹⁴C) 18:33] fatty acids in adult eastern oysters,Crassostrea virginica Gmelin (spawned from parents obtained in 1986 from Mobjack Bay, Virginia, USA), and the influence of temperature on these processes. InC. virginica, incorporation of injected palmitic (16:0) and linolenic (18:33) acids was increased when oysters which had been grown in warm water (22 to 23°C) were transfered to cold water (5 to 7°C) for 8 to 18 d. Incorporation of linoleic acid (18:26) was unchanged under these conditions. The changes in concentration may have been linked to depression of metabolism in these oysters, in particular that of 16:0, which was reduced by 90%. Oxidation of incorporated fatty acids was much higher in warm than in cold water. Cold-temperature conditioning ofC. virginica altered the distribution of fatty acids among the neutral and polar lipid fractions. Long-term exposure to cold water increased the proportion of fatty acids in the polar fraction, which may be related to maintenance of membrane fluidity. Short-term exposure to cold water had the opposite effect, which may be due to increased energy requirements as the oyster adapts to new conditions. Reutilization of¹⁴C-acyl groups demonstrated de novo synthesis of 16:0 and 18:0 fatty acids. Only limited elongation and no desaturation of the administered fatty acids was observed.     

Persistence of atrazine impact on aneuploidy in Pacific oysters,<Emphasis Type="Italic"> Crassostrea gigas</Emphasis>

Widespread use of the herbicide atrazine has incited much research on its toxicity in aquatic systems, where it is routinely detected due to runoff from cultivated fields. Moreover, the determination of the genotoxic effect of such pollutants in the marine environment has become a major requirement for ecosystem protection. In the Pacific oyster, Crassostrea gigas, hypodiploid aneuploid cells have regularly been reported. There is a negative correlation between this phenomenon and growth, as well as evidence for a genetic basis. A positive relationship between atrazine and aneuploidy has previously been demonstrated in C. gigas adults and juveniles. To evaluate the persistence of this impact, our study examined the offspring of the same adult population previously treated with different atrazine doses (10 µg l^–1, representing a peak value found in a polluted environment and 100 µg l^–1), and a seawater control. We observed that these offspring exhibited significantly higher aneuploidy levels when their parents had been exposed to atrazine (14.9–16.9% in comparison with the control where the levels ranged from 11.4% to 12.8%). In addition, the present study examined the aneuploidy level of a sample of juveniles, previously exposed for 3.5 months to the same doses of atrazine, then transferred to non-polluted conditions for an additional period of 2.5 months; this aneuploidy level remained significantly different between the treatments applied. These results demonstrate the persistence of an atrazine impact on Pacific oyster aneuploidy in time, within and between generations, indicating that this widely used compound may represent an important factor causing at least medium-term damage to genetic material.Communicated by S.A. Poulet, Roscoff     

Contrasting pattern of mitochondrial population diversity between an estuarine bivalve, the Kumamoto oyster Crassostrea sikamea, and the closely related Pacific oyster C. gigas

The Kumamoto oyster (Crassostrea sikamea) shows a spatially restricted distribution, favoring estuarine tideland environment. On the other hand, the Pacific oyster (C. gigas) has a broader range of habitat. The present study compared the mitochondrial population structure between the two closely related species. For accurate species identification of oysters sampled from Japanese and East Asian continental coasts, we performed sequencing analysis of the mitochondrial DNA (mtDNA) and PCR-RFLP assay of the first internal transcribed spacer of nuclear rRNA genes. Then, we estimated the extent of population differentiation within each of C. sikamea and C. gigas based on the mtDNA data. Few haplotypes were shared among the sites of sampling in C. sikamea, which contrasted with an extensive haplotype sharing among C. gigas samples. We discuss the mechanisms of elevated population differentiation observed in C. sikamea in light of the ecology and the ancient ocean geography around the present-day habitats.     

The swimming kinematics of larval Atlantic cod, Gadus morhua L., are resilient to elevated seawater pCO2

Kinematics of swimming behavior of larval Atlantic cod, aged 12 and 27 days post-hatch (dph) and cultured under three pCO₂ conditions (control-370, medium-1800, and high-4200 μatm) from March to May 2010, were extracted from swim path recordings obtained using silhouette video photography. The swim paths were analyzed for swim duration, distance and speed, stop duration, and horizontal and vertical turn angles to determine whether elevated seawater pCO₂—at beyond near-future ocean acidification levels—affects the swimming kinematics of Atlantic cod larvae. There were no significant differences in most of the variables tested: the swimming kinematics of Atlantic cod larvae at 12 and 27 dph were highly resilient to extremely elevated pCO₂ levels. Nonetheless, cod larvae cultured at the highest pCO₂ concentration displayed vertical turn angles that were more restricted (median turn angle, 15°) than larvae in the control (19°) and medium (19°) treatments at 12 dph (but not at 27 dph). Significant reduction in the stop duration of cod larvae from the high treatment (median stop duration, 0.28 s) was also observed compared to the larvae from the control group (0.32 s) at 27 dph (but not at 12 dph). The functional and ecological significance of these subtle differences are unclear and, therefore, require further investigation in order to determine whether they are ecologically relevant or spurious.     

Effects of elevated pCO2 and the effect of parent acclimation on development in the tropical Pacific sea urchin Echinometra mathaei

Effects of acclimation to projected near-future ocean acidification (OA) conditions on physiology, reproduction and development were investigated in the tropical sea urchin Echinometra mathaei. Following 6 weeks in control or one of the three elevated pCO₂ (pH_NIST 7.5–8.1; pCO₂ ~485–1,770 μatm) conditions, adult urchins exhibited a slight decline of growth in low pH treatments and moderately reduced respiration at intermediate levels. At 7 weeks, gametes from adults were used to produce larvae that were reared in their respective parental treatments. To assess whether larvae from acclimated parents are more resilient to elevated pCO₂ than those not acclimated, larvae from control animals were also reared in the elevated pCO₂ treatments. There was no difference in female ‘spawnability’ and oocyte size between treatments, but male spawning ability was reduced in increased pCO₂ conditions. In elevated pCO₂ treatments, the percentage of normal larvae and larval size decreased in the progeny of control- and elevated pCO₂-acclimated parents, and arm asymmetry increased. Thus, acclimation of the parents did not make the progeny more resilient or sensitive to OA effects. Negative effects of increased pCO₂ on reproduction and development may impact on recruitment and population maintenance of this species.     

Feeding current characteristics of three morphologically different bivalve suspension feeders, Crassostrea gigas, Mytilus edulis and Cerastoderma edule, in relation to food competition

Introduced Pacific oysters (Crassostrea gigas) have shown rapid expansion in the Oosterschelde estuary, while stocks of native bivalves declined slightly or remained stable. This indicates that they might have an advantage over native bivalve filter feeders. Hence, at the scale of individual bivalves, we studied whether this advantage occurs in optimizing food intake over native bivalves. We investigated feeding current characteristics, in which potential differences may ultimately lead to a differential food intake. We compared feeding currents of the invasive epibenthic non-siphonate Pacific oyster to those of two native bivalve suspension feeders: the epibenthic siphonate blue mussel Mytilus edulis and the endobenthic siphonate common cockle Cerastoderma edule. Inhalant flow fields were studied empirically using digital particle image velocimetry and particle tracking velocimetry. Exhalant jet speeds were modelled for a range of exhalant-aperture cross-sectional areas as determined in the laboratory and a range of filtration rates derived from literature. Significant differences were found in inhalant and exhalant current velocities and properties of the inhalant flow field (acceleration and distance of influence). At comparable body weight, inhalant current velocities were lower in C. gigas than in the other species. Modelled exhalant jets were higher in C. gigas, but oriented horizontally instead of vertically as in the other species. Despite these significant differences and apparent morphological differences between the three species, absolute differences in feeding current characteristics were small and are not expected to lead to significant differences in feeding efficiency.     

Geographic variation in statolith trace elements of the Humboldt squid, Dosidicus gigas, in high seas of Eastern Pacific Ocean

We applied the solution-based inductively coupled plasma mass spectrometry to quantify trace elements in statoliths of Humboldt squid, Dosidicus gigas, collected from the high seas off Chile, Peru, and Costa Rica by Chinese squid jigging vessels during 2007–2009. All squid samples were aged and their spawning dates were back-calculated based on daily increments in statoliths. The most abundant trace elements in the whole statolith were Ca and Sr followed by other elements in the order of Fe, Mg, Zn, Ba, Cu, Mn, Ni, Al, Cr, Co, and U. Significant differences in Mn and Sr were found among samples from the three regions. Sr, Ni, Mn, and Co contributed significantly to the discrimination among the regions, with Co responsible for explaining most of the variation, followed by Ni, Mn, and Sr. Squid from the high seas off Costa Rica could be separated from those off Peru and Chile mostly due to the differences in Ni, Sr, and Co, while samples off Peru and Chile could be distinguished mainly because of differences in Mn and Co. Discriminant function analysis suggested that the overall cross-validated classification rate was 85.6 % with samples off Chile having the highest correct identification rates and samples off Costa Rica having the highest false classification rates. Significant positive relationships were found between sea surface temperature (SST) and Cr/Ca, Mn/Ca, and U/Ca, and there was a negative relationship between SST and Cu/Ca, Sr/Ca. This study suggests that the spatial difference in trace elements of statolith can be used to separate geographic populations of D. gigas and that elements having significant relationships with SST can be considered as natural indicators of ambient temperature.     

The false limpet Siphonaria gigas, a simultaneous hermaphrodite, lives in pairs in rock fissures on the Pacific coast of Panama

The pulmonate limpet Siphonaria gigas, a simultaneous hermaphrodite, lives in the mid- to upper-intertidal zone on rocky shores in the tropical Eastern Pacific. Samples along five transects taken in June–July, 2004, on Culebra Point (8º54′N to 79º31′W), Republic of Panama, showed that 71 % of the population occurred in fissures, a significant preference for this habitat. Of 200 adults in 27 fissures, 150 lived side by side in pairs with their shells touching or nearly so, a significant deviation from the number of pairs expected given a random arrangement. Pair frequency did not increase with limpet density suggesting pairing was not an incidental consequence of crowding. Pair living was unknown in the genus Siphonaria, and is very uncommon among simultaneous hermaphrodites. Reproductive synchrony and restrictions on movement due to predation and environmental stress may limit opportunities for encountering and mating with multiple partners favoring pair living in S. gigas.