Identification of Mytilus spp. and Pecten maximus in Irish Waters by Standard PCR of the 18S rDNA Gene and Multiplex PCR of the 16S rDNA Gene

Two molecular protocols for the identification of mussel and scallop have been developed using specific primers targeting the mitochondrial 16S ribosomal DNA gene and the nuclear 18S ribosomal DNA gene. Primers for the mitochondrial 16S ribosomal DNA gene in multiplex polymerase chain reaction (PCR) protocols yielded diagnostic DNA fragments for the mussels Mytilus edulis, Mytilus galloprovincialis, and the hybrid Mytilus edulis/galloprovincialis (335 bp), the king scallop Pecten maximus (382 bp) and the black scallop Mimachlamys varia (398 bp). DNA from the queen scallop Aequipecten opercularis showed no consistent PCR amplification of the 16S rDNA gene. Primers for the nuclear 18S rDNA gene in standard PCR protocols yielded similar-sized, diagnostic DNA fragments (approx. 190 bp) for the mussels Mytilus edulis, Mytilus galloprovincialis, and the hybrid Mytilus edulis/galloprovincialis, the king scallop Pecten maximus, the black scallop Mimachlamys varia, and the queen scallop Aequipecten opercularis. Both protocols have been tested with Mytilus spp., P. maximus, and 6 other bivalve species from a wide range of locations in Irish and European waters. Cross reaction of the specific primers with DNA template from any of the 6 other bivalve species was not observed. Rapid DNA extraction using FTA Card technology and the16S rDNA primers allowed for the detection of at least 10 mussel larvae in a subsample of natural plankton.     

Maerl grounds provide both refuge and high growth potential for juvenile queen scallops (Aequipecten opercularis L.)

Human damage to biogenic substrata such as maerl has been receiving increasing attention recently. Maerl forms highly biodiverse and heterogeneous habitats composed of loose-lying coralline red algae, which fulfil nursery area prerequisites for queen scallops (Aequipecten opercularis) and other invertebrates. The benefits obtained by queen scallops utilising maerl were poorly understood, so we used both laboratory predation and field tethering experiments to investigate the refuge and growth potential provided by pristine live maerl (PLM) grounds over other common substrata. In aquaria, more juvenile queen scallops (<35 mm shell height) survived on PLM than on gravel substrata in the presence of the crab Carcinus maenas or the starfish Asterias rubens. Field tethering experiments indicated similar survivorship of juvenile queen scallops on PLM than gravel; additionally, their growth rates were similar on both substrata. PLM allows scallops to seek refuge from predators and position themselves to optimise their food supply. Other bivalve refugia have been shown to provide poor food supply as a consequence of their high heterogeneity, yet maerl grounds provide a ‘win-win’ scallop nursery area coupling refuge availability with high food supply.     

Margolisiella islandica sp. nov. (Apicomplexa: Eimeridae) infecting Iceland scallop Chlamys islandica (Müller, 1776) in Icelandic waters

Wild Iceland scallops Chlamys islandica from an Icelandic bay were examined for parasites. Queen scallops Aequipecten opercularis from the Faroe Islands and king scallops Pecten maximus and queen scallops from Scottish waters were also examined. Observations revealed heavy infections of eimeriorine parasites in 95–100% of C. islandica but not the other scallop species. All life stages in the apicomplexan reproduction phases, i.e. merogony, gametogony and sporogony, were present. Trophozoites and meronts were common within endothelial cells of the heart’s auricle and two generations of free merozoites were frequently seen in great numbers in the haemolymph. Gamonts at various developmental stages were also abundant, most frequently free in the haemolymph. Macrogamonts were much more numerous than microgamonts. Oocysts were exclusively in the haemolymph; live mature oocysts contained numerous (>500) densely packed pairs of sporozoites forming sporocysts.Analysis of the 18S ribosomal DNA revealed that the parasite from C. islandica is most similar (97.7% identity) to an unidentified apicomplexan isolated from the haemolymph of the giant clam, Tridacna crocea, from Japan. Phylogenetic analyses showed that the novel sequence consistently grouped with the Tridacna sequence which formed a robust sister clade to the rhytidocystid group.We propose the name Margolisiella islandica sp. nov., referring to both type host and type locality.     

Attachment of the juvenile queen scallop (Aequipecten opercularis (L.)) to maerl in mesocosm conditions; juvenile habitat selection

Substratum un-acclimated juvenile queen scallops (Aequipecten opercularis) (<18 mm and 18-30 mm shell height) were released in equal numbers onto pristine live maerl (PLM), impacted dead maerl (IDM), gravel and sand in choice chambers. Their habitat selection was monitored over a 4-day period in control and predator treatments (utilising Asterias rubens L. and Carcinus maenas (L.)). Microhabitat use of PLM by juvenile queen scallops and the presence of cues in live maerl were also investigated.In control and predator treatments juvenile queen scallops were observed to attach preferentially to PLM than IDM, gravel or sand. Juvenile queen scallops were observed to maintain a more exposed attachment site in the absence of predators but sought refuge within and between maerl nodules in the presence of both predators. Smaller queen scallops (<18 mm shell height) were more efficient at utilising maerl thalli as a refuge. Juvenile A. opercularis showed hierarchical cue responses mediated by predator presence, i.e. responding favourably to a factor associated with live maerl presence irrespective of heterogeneity in the absence of predators but favourably to higher maerl heterogeneity in their presence. If they also preferentially attach to PLM in the field, at some sites where PLM grounds cover large areas, they may thus be considered to constitute ‘nursery areas’. Habitat attachment preference appears to be predetermined and not a result of localised predator avoidance; however, habitat usage changes in the presence of predators. Maerl beds have been shown to be easily damaged by scallop dredging in Scotland and if such nursery areas are being destroyed extensively in the field, this could damage recruitment to localised adult populations.     

Environmental variability at the phosphoglucose isomerase locus in the genus Chlamys

Two geographically and reproductively isolated populations each of Chlamys opercularis and of Chlamys varia are compared with respect to polymorphism at the phosphoglucose isomerase (PGI) locus. One population of C. varia is further sub-divided into sub-littoral and inter-tidal samples. From the results it is inferred that PGI is a dimeric molecule, controlled by five alleles in C. varia and by at least four alleles in C. opercularis. acting at a single locus. The alleles recognised for C. opercularis have identical electrophoretic mobility as four of those seen in C. varia. The two populations of C. varia are remarkably similar, as were their habitats. However, the sub-littoral sample does show a lesser degree of homozygosity than the inter-tidal sample of the same population. The two populations of C. opercularis differ significantly and represent two separate and identifiable ‘races’. C. varia exhibits a much greater degree of genetic variability than the deep sub-littoral species, C. opercularis indicating that a more stable environment regulates a more monomorphic species.     

Interspecific comparison of Cd bioaccumulation in European Pectinidae (Chlamys varia and Pecten maximus)

The uptake and loss kinetics of Cd were determined in two species of scallops from the European coasts, the variegated scallop Chlamys varia and the king scallop Pecten maximus, following exposures via seawater, phytoplankton and sediment using highly sensitive radiotracer techniques (¹⁰⁹Cd). Results indicate that, for seawater and dietary pathways, C. varia displays higher bioaccumulation capacities in terms of uptake rate from water and fraction absorbed from ingested food (assimilation efficiency) than Pecten maximus. Regarding sediment exposure, P. maximus displayed low steady-state Cd transfer factor (TF_SS < 1); however, once incorporated, a very large part of Cd transferred from sediment (92%) was strongly retained within P. maximus tissues.Both species showed a high retention capacity for Cd (biological half-life, T_b1/2 > 4 months), suggesting efficient mechanisms of detoxification and storage in both species. The digestive gland was found to be the main storage organ of Cd in the two scallops regardless of the exposure pathway. However, Cd was stored differently within this organ according to the species considered: 40% of the total Cd was found in the soluble cellular fraction in C. varia whereas this soluble fraction reached 80% for P. maximus. This suggests that the two species displayed different Cd detoxification/storage mechanisms.Finally, the present study has determined the relative contribution of the different exposure pathways to global Cd bioaccumulation for the two scallop species. Results clearly show that for both species, food constitutes the major accumulation pathway, contributing for > 99% and 84% of the global Cd bioaccumulation in C. varia and P. maximus, respectively. This work confirms the previous assumption, derived from a bibliographic overview, that dietary pathway plays a prevalent role in metal bioaccumulation in Pectinidae.     

Karyotype and Chromosomal Location of 18S–28S and 5S Ribosomal DNA in the Scallops Pecten maximus and Mimachlamys varia (Bivalvia: Pectinidae)

This work describes the karyotype and chromosomal location of the ribosomal DNA (rDNA) of Pecten maximus and Mimachlamys varia, two commercial scallop species from Europe. According to the chromosome centromeric index values found, the karyotype of P. maximus is composed of 1 metacentric, 2 metacentric–submetacentric, 1 telocentric–subtelocentric and 15 telocentric pairs, and that of M. varia of 4 metacentric, 2 subtelocentric–submetacentric, 9 subtelocentric, 3 subtelocentric–telocentric and 1 telocentric–subtelocentric pairs. In P. maximus, 18S-28S rDNA was located by FISH on a metacentric–submetacentric pair, and in M. varia on a subtelocentric–submetacentric pair using both silver staining and FISH. PCR amplification of the 5S rDNA unit yielded a single product of about 460 bp (P. maximus) and 450 bp (M. varia), that used as probe revealed a 5S rDNA site on a telocentric pair in P. maximus and a subtelocentric pair in M. varia. Two-color FISH or sequential silver staining of 5S rDNA-FISH-metaphases corroborated that the two gene families are located on different chromosomes in both species. A comparative analysis of the data allowed the inference of karyotypic relationships within scallops.     

Characterization of 12 single nucleotide polymorphisms in weathervane scallop

We describe 27 single nucleotide polymorphisms (SNPs) in a commercially important bivalve, the weathervane scallop (Patinopecten caurinus), identified using a targeted‐gene approach. We further characterize 12 of these using 5′‐nuclease and allele‐specific PCR assays. Polymorphisms were identified in both mitochondrial and nuclear genes. These are the first SNPs developed for delineating population structure in the weathervane scallop and will provide a useful complement to currently available genetic markers.     

Seasonal swimming behaviour in the queen scallop (Aequipecten opercularis) and its effect on dredge fisheries

Modified dredges were used on Aequipecten opercularis (queen scallop) fishing grounds off the Isle of Man in the north Irish Sea to determine seasonal variability in swimming behaviour in queen scallops and its effect on dredge fisheries. Scallops, which evaded dredge capture by swimming up into the water column, were captured by a specially designed net deployed above the dredge gear. The gear was used over a 20-month period and the number and size distribution of queen scallops captured in both the net and dredges were recorded. A subsample of captured scallops was maintained in running seawater tanks and used to assess two components of swimming behaviour, the time taken to respond to stimuli and the number of valve adductions carried out until exhaustion. There was considerable seasonal variability in the proportion of queen scallops that avoided dredge capture by swimming. Very low numbers of queen scallops were found in the net during the winter and spring. Net capture increased in June, and was high throughout the summer and autumn when up to 42% of scallops greater than 55 mm in height, captured in dredges, swam over the dredge mouths. There was a strong positive correlation between seawater temperature and proportion of captured scallops found in the net. Swimming experiments in the laboratory indicated that the observed seasonal variability in net capture was probably a result of changes in the time taken to respond to stimuli, rather than changes in the ability to perform a large number of valve adductions. There was no relationship between reproductive state and the proportion of actively swimming scallops. Analyses of size distributions indicated limited swimming activity in the largest scallops but no effect of epifaunal load was found on the ability of scallops to avoid dredge capture.     

Development of an Argopecten-Specific 18S rRNA Targeted Genetic Probe

Comparison of 18S ribosomal RNA gene sequences between diverse bivalve species, including eight scallop species, allowed the design of an 18S rRNA targeted oligonucleotide probe (BS-1364) that was specific for scallops belonging to the genus Argopecten (bay and calico scallops). The high sequence similarity of the 18S rRNA gene between Argopecten irradians and Argopecten gibbus (98.8%) prevented the design of an A. irradians species-specific probe. Hybridization studies using amplified 18S rDNA from a diverse collection of bivalve species demonstrated that the specificity of the digoxygenin-labeled probe was consistent with the predicted specificity indicated by sequence comparison. Hybridization studies using laboratory-spawned bay scallop veligers indicated that a single veliger could be detected by probe hybridization in a blot format, and that probe hybridization signal was proportional (r ²= .99) to the abundance of veligers. Methods for rRNA extraction and blotting were developed that allowed bay scallop veligers to be specifically and quantitatively identified in natural plankton samples. Preliminary studies conducted in Tampa Bay, Florida, suggest that introduced scallops can successfully spawn and produce veligers under in situ conditions. The Argopecten-specific probe and methods developed in this study provide the means to study the production and fate of bay scallop larvae in nature and provide evidence that scallops introduced into Tampa Bay have the potential for successful reproduction and enhancement of scallop stocks. Received January 25, 1999; accepted May 7, 1999.     

Polymorphic microsatellite loci isolated from the great scallop,Pecten maximus (Bivalvia: Pectinidae)

We isolated and characterized nine polymorphic microsatellite loci from the great scallop (Pecten maximus) as part of an attempt to better understand the population–genetic structure of this species. Levels of genetic diversity were assessed in 32 scallops collected from the coastal waters of Port Erin Bay, Isle of Man. The number of alleles ranged from two to 29 and observed and expected heterozygosities varied between 0.031 to 0.906 and 0.062 to 0.973, respectively. Two loci showed significant (P < 0.05) heterozygote deficits, likely because of null (nonamplifying) alleles; one pair of loci was in linkage disequilibrium.     

Development of Microsatellite Markers for Japanese Scallop (Mizuhopecten yessoensis) and Their Application to a Population Genetic Study

Abstract The Japanese scallop (Mizuhopecten yessoensis) is one of the main fishery products in Japan, but with the expansion of culture operations of the Japanese scallop, various problems have been encountered including high mortality, poor growth, poor seed production, and so on. Moreover, there is concern that many years of cultivation may have affected the genetic structure of the scallop population. To approach these problems and concerns, we developed microsatellite markers as a molecular tool for population genetic studies. By using 4 microsatellite markers as well as a mitochondrial marker, we investigated the genetic structure of samples from the islands of Hokkaido (14 populations) and Honshu (Tohoku, 3 populations) in Japan, and south Primorye (4 populations) in Russia. All the populations sampled had high genetic diversity (average expected heterozygosity, 0.7011 to 0.7622; haplotype diversity, 0.6090 to 0.8848), and almost all showed a tendency of homozygote excess, which was significant in 2 populations. Hierarchical analysis of molecular variance tests based on the microsatellite and mitochondrial markers indicated that the 3 geographic regions were genetically divergent from one another, with little evidence of divergence within regions. Homogeneity in allele frequency distributions between natural and cultured scallops and allele frequency stability over a period of 2 decades indicated that the culturing operations have probably not had a substantial effect on the genetic structure of the populations.     

Habitat Characteristics Predicting Distribution and Abundance Patterns of Scallops in D’Entrecasteaux Channel,Tasmania

Habitat characteristics greatly influence the patterns of distribution and abundance in scallops, providing structure for the settlement of spat and influencing predation risk and rates of survival. Establishing scallop-habitat relationships is relevant to understanding the ecological processes that regulate scallop populations and to managing critical habitats. This information is particularly relevant for the D’Entrecasteaux Channel, south-eastern Tasmania (147.335 W, 43.220 S), a region that has supported significant but highly variable scallop production over many years, including protracted periods of stock collapse. Three species of scallops are present in the region; the commercial scallop Pecten fumatus, the queen scallop Equichlamys bifrons, and the doughboy scallop Mimachlamys asperrima. We used dive surveys and Generalized Additive Modelling to examine the relationship between the distribution and abundance patterns of each species and associated habitat characteristics. The aggregated distribution of each species could be predicted as a function of sediment type and species-specific habitat structural components. While P. fumatus was strongly associated with finer sediments and E. bifrons with coarse grain sediments, M. asperrima had a less selective association, possibly related to its ability to attach on a wide range of substrates. Other habitat characteristics explaining P. fumatus abundance were depth, Asterias amurensis abundance, shell and macroalgae cover. Equichlamys bifrons was strongly associated with macroalgae and seagrass cover, whereas M. asperrima abundance was greatly explained by sponge cover. The models define a set of relationships from which plausible hypotheses can be developed. We propose that these relationships are mediated by predation pressure as well as the specific behavioural characteristics of each species. The findings also highlight the specific habitat characteristics that are relevant for spatial management and habitat restoration plans.     

Effects of sponge and barnacle encrustation on survival of the scallop <Emphasis Type="Italic">Chlamys hastata</Emphasis>

The scallop Chlamys hastata frequently carries epibionts such as sponges and barnacles on its shells. Although the scallop-sponge relationship has been characterized as a mutualism, little is known about the scallop-barnacle relationship. This study investigated the effects of sponge and barnacle encrustation on the ability of C. hastata to avoid predation by the sea star Pycnopodia helianthoides. In feeding trials, P. helianthoides caught and consumed significantly more barnacle-encrusted scallops (7.7 ± 0.8 out of 20 scallops) than scallops encrusted by either of the sponges Myxilla incrustans (4.1 ± 0.9) or Mycale adhaerens (3.0 ± 0.5). Epibiont-free scallops (5.7 ± 0.5) formed an intermediate treatment between barnacle-encrusted and sponge-encrusted scallops. Possible mechanisms by which the sponges protected the scallops were investigated in two ways: two feeding trials were videotaped to allow qualitative analysis of sea star and scallop behavior and sea star feeding responses to scallop and sponge homogenates were determined to investigate if sea stars accept scallops and sponges as prey. Sea stars displayed positive feeding responses to scallop puree 97.5% ± 1.6 of the time while only displaying positive responses to Mycale adhaerens homogenate 4.4% ± 2.0 of the time and to Myxilla incrustans homogenate 4.4% ± 2.9 of the time. The videotaped feeding trials indicated that interference with tube feet adhesion by the sponge deterred predation. Observations of both sea stars that were videotaped showed that neither avoided trying to capture sponge-encrusted scallops, and at no time was a captured scallop willingly released by the sea stars. Thus, it appears that sponges provide tactile-mechanical protection and possibly chemical or tactile camouflage in this predator/prey relationship. Finally, the effects of sponge encrustation on barnacle settlement were determined. Field experiments showed that barnacle larvae settled more frequently on epibiont-free scallops than on those with either of the two sponges, potentially protecting the scallops from an epibiont that increases the scallop’s susceptibility to predation. Handling editor: K. Martens     

Oceanographic variation influences spatial genomic structure in the sea scallop,Placopecten magellanicus

Environmental factors can influence diversity and population structure in marine species and accurate understanding of this influence can both improve fisheries management and help predict responses to environmental change. We used 7163 SNPs derived from restriction site‐associated DNA sequencing genotyped in 245 individuals of the economically important sea scallop, Placopecten magellanicus, to evaluate the correlations between oceanographic variation and a previously identified latitudinal genomic cline. Sea scallops span a broad latitudinal area (>10 degrees), and we hypothesized that climatic variation significantly drives clinal trends in allele frequency. Using a large environmental dataset, including temperature, salinity, chlorophyll a, and nutrient concentrations, we identified a suite of SNPs (285–621, depending on analysis and environmental dataset) potentially under selection through correlations with environmental variation. Principal components analysis of different outlier SNPs and environmental datasets revealed similar northern and southern clusters, with significant associations between the first axes of each (R²_adj = .66–.79). Multivariate redundancy analysis of outlier SNPs and the environmental principal components indicated that environmental factors explained more than 32% of the variance. Similarly, multiple linear regressions and random‐forest analysis identified winter average and minimum ocean temperatures as significant parameters in the link between genetic and environmental variation. This work indicates that oceanographic variation is associated with the observed genomic cline in this species and that seasonal periods of extreme cold may restrict gene flow along a latitudinal gradient in this marine benthic bivalve. Incorporating this finding into management may improve accuracy of management strategies and future predictions.     

A short note on heavy cunjevoi growth observed on commercial scallops (Pecten fumatus)

ABSTRACT

Commercial scallops (Pecten fumatus) were collected from Bass Strait, Australia from 41 dredge tows. Of these, four dredges undertaken in February 2016 at 46 m depth returned scallops that were covered by ascidians of the Pyura stolonifera species complex, commonly known as cunjevoi. There were no obvious signs of immediate ill health in the scallops, with meat being assessed as normal quality and all scallops requiring force to separate the shells. Ascidian-encrusted scallops were significantly smaller, and previous tows in the same location ten and eight months earlier returned far fewer scallops with clean shells and no signs of ascidians. This suggests that both both scallop and ascidian recruitment and growth occurred during the period between sampling events. Future research combining laboratory experiments and field observations is recommended to understand this relationship and its potential impacts on scallop populations.     

Genetic structure of Micromeria (Lamiaceae) in Tenerife,the imprint of geological history and hybridization on within‐island diversification

Geological history of oceanic islands can have a profound effect on the evolutionary history of insular flora, especially in complex islands such as Tenerife in the Canary Islands. Tenerife results from the secondary connection of three paleo‐islands by a central volcano, and other geological events that further shaped it. This geological history has been shown to influence the phylogenetic history of several taxa, including genus Micromeria (Lamiaceae). Screening 15 microsatellite markers in 289 individuals representing the eight species of Micromeria present in Tenerife, this study aims to assess the genetic diversity and structure of these species and its relation with the geological events on the island. In addition, we evaluate the extent of hybridization among species and discuss its influence on the speciation process. We found that the species restricted to the paleo‐islands present lower levels of genetic diversity but the highest levels of genetic differentiation suggesting that their ranges might have contracted over time. The two most widespread species in the island, M. hyssopifolia and M. varia, present the highest genetic diversity levels and a genetic structure that seems correlated with the geological composition of the island. Samples from M. hyssopifolia from the oldest paleo‐island, Adeje, appear as distinct while samples from M. varia segregate into two main clusters corresponding to the paleo‐islands of Anaga and Teno. Evidence of hybridization and intraspecific migration between species was found. We argue that species boundaries would be retained despite hybridization in response to the habitat's specific conditions causing postzygotic isolation and preserving morphological differentiation.     

Damage to the eyes of the bivalve Chlamys opercularis caused by the ciliate Licnophora auerbachii

The majority of specimens of the scallop Chlamys opercularis found in Strangford Lough, County Down, Ireland, are infected with a rare and exotic ciliate Licnophora auerbachii. The ciliate is found predominantly on the eyes of the scallop which (together with numerous tentacles) are borne by the middle marginal folds of the mantle. The ciliates are found attached to both the clear cornea covering the lens and also the surrounding pigmented iris, their large food vacuoles often containing black consistently sized granules, particles derived from the pigmented epithelial cells of the scallop eye. These cells are abraded from the surface of the iris and become detached due to the action of the basal disc of L. auerbachii during its clamping and locomotory activities on the eye surface. Scanning electron microscope observations show that the foot region of the ciliate exerts both a suction and abrasive action on the eye and leaves a distinct “footprint” on the eye surface in the form of a saucer-shaped depression. In many of the heavily infected scallops there was evidence of damage to the eyes with consequent loss of pigment and in some instances surface distortion of the eye itself.     

Development and Linkage Analysis of 104 New Microsatellite Markers for Bay Scallop (Argopecten irradians)

For genetic analysis and linkage mapping of bay scallop (Argopecten irradians), a set of 120 novel simple sequence repeat markers were developed from microsatellite-enriched libraries and expressed sequence tags. An inter-subspecies hybrid bay scallop family (CC5) of 46 progeny was analyzed as the reference population to confirm polymorphism and test the segregation patterns of these loci. A total of 104 microsatellite markers were polymorphic in the reference family, among which 36 in female, 28 in male, and 40 in both parents, respectively. Linkage analysis allowed mapping these markers to 15 linkage groups, which is close to the haploid chromosome number of bay scallop (n = 16). Analysis of the 40 markers segregating in both parents showed a higher recombination rate in the female parent, with the average of female-to-male recombination ratio of 1.09:1 between linked pairs of markers. When null alleles were considered, there were 17 loci showing segregation distortion at the 5% significance level using the chi-square test. The microsatellite markers developed in this study provide a useful resource for future linkage mapping and quantitative loci analysis in A. irradians.