Nematocyst complements of nudibranchs in the genus <Emphasis Type="Italic">Flabellina</Emphasis> in the Gulf of Maine and the effect of diet manipulations on the cnidom of <Emphasis Type="Italic">Flabellina verrucosa</Emphasis>

Aeolid nudibranchs maintain functional nematocysts, which are sequestered from the nudibranchs’ cnidarian prey and provide protection against predators. Some species exhibit extensive variation in incorporated nematocysts, while others maintain a limited number of types. This study examines the apparent diversity in uptake and patterns of nematocyst incorporation among related species. Nematocyst complements were described for four Gulf of Maine nudibranch species in the genus Flabellina exhibiting a variety of feeding strategies and prey specificities. Diet manipulations were performed to examine the response to changing nematocyst availability using a generalist consumer, Flabellina verrucosa, to assess nematocyst uptake based on diet. The flabellinid species examined exhibited significant differences in nematocyst incorporation, reflecting differences in their specificity as predators and nematocyst types available in their natural prey. The nematocyst complement of F. verrucosa was the most variable and differed among collection regions. When diet was manipulated, nematocyst uptake depended on the prey the nudibranchs consumed, but when offered a variety of prey, F. verrucosa selectively preferred nematocysts from scyphistomae. The observed variation in nematocyst uptake among species and regions probably relates to environmental disparities among populations.     

Genetic differentiation between morphotypes of the marine sponge Suberites ficus (Demospongiae: Hadromerida)

Sponges of three morphotypes of Suberites ficus (Johnston, 1842) were collected during February and March 1985 off the south-west of the Isle of Man, and were compared by using spicule size distributions and genetic allele frequencies of isozyme loci. The populations did not show any significant differences of spicule size or type, but could be easily differentiated into three separate species based on isozyme patterns. Samples of pale orange S. ficus growing on gastropod shells inhabited by hermit crabs (Pagurus spp.) were reproductively isolated from the redorange and the pale yellow colour morphs encrusting the bivalve Chlamys opercularis. These latter two colour morphs were genetically similar, but significant differences were observed at two of the 19 gene loci assayed. All the sponges studied were sympatric, and therefore the genetic differences, indicating reproductive isolation, are strong evidence for separate gene pools and, hence, that they are different species. The genetic identity between the two colour morphs of S. ficus on C. opercularis shells was 0.977, whilst between each of these and S. ficus on hermit crabs it was about 0.65. In all three species genetic variability was high, with mean expected and observed heterozygosity values per locus ranging from 0.17 to 0.36.     

Variation in the genetic composition of coral (Pocillopora damicornis and Acropora palifera) populations from different reef habitats

The genetic structure of populations of the corals Pocillopora damicornis and Acropora palifera was examined in three habitats at One Tree Island during March and April 1993, using electrophoretically detectable variation at six allozyme loci. There were significant genetic differences among populations of P. damicornis within each of the reef crest, lagoon and microatoll habitats. The level of differentiation among populations was similar in each of the habitats. Differences between populations of P. damicornis from lagoon and microatolls were no greater than that within habitats, but genetic differentiation of these from crest populations was much higher. There was no difference in the genetic composition of A. palifera populations within or between the lagoon and microatolls, the only habitats where this species was found. Both coral species had observed:expected (G _O:G_E) genotypic diversity rations >0.80, indicating predominantly sexual reproduction. These data, the high genotype diversity and general conformance of genotype frequencies to those expected under conditions of Hardy-Weinberg, suggested panmixis at each site. The high degree of sexual reproduction in the P. damicornis populations is unusual for a species where asexual reproduction has been the dominant mode of reproduction reported to date. Gene flow in both species was considerable between the lagoon and the closed microatolls. The genetic differences between populations of P. damicornis in these habitats and the reef crest may reflect the relative isolation of all populations within the closed One Tree Lagoon from those outside. However, local currents appear to offer effective means of dispersal between the habitats, suggesting that the genetic differences result from natural selection in the different environments within One Tree Lagoon and the reef crest.     

Genetic evidence for the occurrence of a cryptic species with the littoral nemerteans Lineus ruber and L. viridis (Nemertea: Anopla)

Lineus ruber and L. viridis are common intertidal heteronemerteans found on the coasts of northern Europe and the east coast of North America. The two species resemble each other morphologically and were synonymised for many years prior to being separated on the basis of larval development. In this study, specimens of L. ruber and L. viridis were collected from sites along the west and southwest coast of Britain, northern France and the east coast of North America. The external morphology and allele frequencies of isozymes of the two species were compared from all sites. The external morphology of L. ruber and L. viridis was similar but they could generally be separated by colour. Allele frequencies for up to 13 enzyme loci between sympatric populations of L. ruber and L. viridis indicated that these two species are genetically very different (Nei's genetic identity=0.090 to 0.083). Allele-frequency data also indicated the presence of a third genetic type occurring sympatrically with populations of both L. ruber and L. viridis on the coasts of Britain and France but not on the coast of North America. Fixed differences in allele frequencies between populations of the third genetic type and sympatric populations of L. ruber and L. viridis were observed across multiple loci. Genetic identity between the third genetic type and sympatric populations of L. ruber and L. viridis were extremely low (Nei's genetic identity =<0.078). Such large genetic differences between populations indicate a barrier to gene flow and reproductive isolation. The aberrant type, which exhibits sufficient morphological variation to prevent individuals being distinguished from those of both L. ruber and L. viridis, therefore represents a separate species. The low genetic identities found in intrageneric comparisons of species found in this study have been found in other studies on nemerteans. They may indicate systematic problems within these groups or other phenomena such as morphological stasis.     

Enzyme activities and allozyme polymorphism in two genetic types (or sibling species) of the genus Marenzelleria (Polychaeta: Spionidae) in Europe

Two populations of Marenzelleria spp. from the North Sea and the Baltic Sea that had already been characterized by allozyme electrophoresis and found genetically different in 1994 were again examined in 1997. The genetic identity (I = 1.0) examined in 1994 and 1997 shows that the structures of the populations of genetic Type I in the North Sea (Ringkobing Fjord) and genetic Type II in the Baltic Sea (Darss–Zingst Bodden chain) are stable. Three diagnostic gene loci that were determined additionally (citrate synthase, one locus; alanine aminotransferase, two loci) support the hypothesis of the existence of two sibling species of the genus Marenzelleria in Europe. In addition, the activities of ten electrophoretically separated enzyme systems were measured in the worms immediately after removal from their habitat and after they had been kept in the laboratory for 2 months. Under both conditions four enzymes showed significant differences between genetic types. The higher activity of the alanine aminotransferase of the North Sea worms (Type I) is discussed in the context of the different preferred salinities of the two Marenzelleria sibling species. Received: 21 January 1999 / Accepted: 13 July 1999     

The Mytilus edulis species complex in southwest England: effects of hybridization and introgression upon interlocus associations and morphometric variation

Loglinear analysis of electrophoretic data from two hybrid Mytilus edulis x galloprovincialis populations in southwest England revealed non-significant associations between genotypes at four allozyme loci, each of which is partially diagnostic for differences between the two taxa. Significant non-random genotypic associations within the context of the non-significant model involved all four assayed loci equally, consistent with their occurrence in a relatively tight linkage group. Multivariate analyses were used to examine electrophoretic variation from the two hybrid populations, and morphometric variation in the hybrid populations and in four allopatric (two M. edulis and two M. galloprovincialis) populations from western Europe While the number of hybrid mussels is high at both sites (22% at Croyde, 53% at Whitsand) the two taxa have largely maintained the genetic differences which exist between them in allopatry. However, morphological differences between the taxa have been eroded for mussels within the hybrid zone, whereas these differences are quite pronounced for mussels from allopatric populations. It is proposed that each taxon within the genus maintains its genetic identity, despite high dispersal potential, widespread hybridization, and high levels of introgression, as a result of adaptation to different environments. The worlwide occurrence of all four Mytilus hybrid zones at ecotones between recognized biogeographical provinces which are characterized by differences in temperature and salinity is consistent with such an interpretation.     

Genetic structure and allozyme variation of sea bass (Dicentrarchus labrax and D. punctatus) in the Mediterranean Sea

This paper reports data on 28 allozyme loci in wild and artificially reared sea bass (Dicentrarchus labrax) samples, originating from either coastal lagoon or marine sites in the Mediterranean Sea. F _ST analysis (θ estimator) indicated strong genetic structuring among populations; around 34% of the overall genetic variation is due to interpopulation variation. Pairwise θ estimates showed that, on average, the degree of genetic structuring was much higher between marine populations than between samples from lagoons. Six polymorphic loci showed differences in allele frequencies between marine and lagoon samples. Multivariate analyses of individual allozymic profiles and of allele frequencies suggested that different arrays of genotypes prevail in lagoons compared to marine samples, particularly at those loci that, on the basis of previous acclimation experiments, had been implicated in adaptation to freshwater. On the other hand, variation at “neutral” allozyme loci reflects to a greater extent the geographic location of populations. Allozyme differentiation was also studied in a D. labrax population from the Portuguese coast. Average genetic distance between this population and the Mediterranean populations was quite high (Nei's D = 0.236) and calls into question the taxonomic status of the Portuguese population. Finally, genetic relationships between D. labrax and D. punctatus were evaluated. Average Nei's D was 0.648, revealing high genetic differentiation between the two species, even for two sympatric populations of these species in Egypt; thus gene flow was not indicated between species. Received: 24 October 1996 / Accepted: 27 November 1996     

A biochemical genetic study of population structure in queen scallop (Chlamys opercularis) stocks in the Northern Irish Sea

Electrophoresis was used to survey a number of enzymes for loci of potential value as stock markers in Chlamys opercularis (L.). Two loci, Pgi and Pgd, were then studied in larger samples from each of two localities around the Isle of Man. One Pgi allele was found to be restricted to a particular year-class in both populations. This was confirmed by subsequent studies of three further populations, in each of which the same allele occurred only in the one year-class. Otherwise, no significant differences in allele frequencies were found between year-classes or between populations in any of the samples used. From the results it is concluded that there is temporal variation in the sources of recruitment of C. opercularis stocks around the Isle of Man. Possible origins of larvae contributing to these stocks are discussed in the light of what is known of current circulation patterns in the northern Irish Sea.     

Allozyme electrophoresis demonstrates that the scleractinian coral Montipora digitata is two species

Allozyme electrophoresis at five loci was used to clarify the species status of two morphs of the scleractinian coral Montipora digitata (Dana, 1846), using specimens collected from three locations off the Queensland coast between October 1991 and April 1993. The two morphs, occurring sympatrically, were distinguished by one fixed gene difference at Locus LT-2 and frequency differences at three other loci, indicating reproductive isolation between them. Populations of both morphs were generally in Hardy-Weinberg equilibrium, suggesting that sexual reproduction does occur. Despite sampling designed to reduce the chance of collecting clonemates, the genetic diversity ratio (G _O:G_E) indicated that asexual reproduction also occurs. This is in agreement with the known modes of reproduction for this species. There was significant genetic differentiation between populations of one of the morphs of M. digitata. This could have been influenced by greater degrees of asexual reproduction in this morph, selection within sites, or variation in larval survival and recruitment patterns. Relatively low F _st (differentiation among populations) values found for a broadcast spawning species during this study are in accordance with the hypothesis that brooded planulae are adapted for rapid settlement, whereas planulae produced by broadcast spawners are adapted for widespread dispersal.     

Genetic evidence for the cryptic species pair, <Emphasis Type="Italic">Lottia digitalis </Emphasis>and <Emphasis Type="Italic">Lottia austrodigitalis</Emphasis> and microhabitat partitioning in sympatry

It has been proposed that the common West Coast limpet, Lottia digitalis, is actually the northern counterpart of a cryptic species duo including, Lottia austrodigitalis. Allele frequency differences between southern and northern populations at two polymorphic enzyme loci provided the basis for this claim. Due to lack of further evidence, L. austrodigitalis is still largely unrecognized in the literature. Seven additional enzyme loci were examined from populations in proposed zones of allopatry and sympatry to determine the existence of L. austrodigitalis as a sibling species to L. digitalis. Significant allele frequency differences were found at five enzyme loci between populations in Laguna Beach, southern California, and Bodega Bay, northern California; strongly supporting the existence of separate species. Both species exhibit two microhabitat morphotypes, a gooseneck barnacle morph in the mid-intertidal zone and a rock morph in the high-intertidal zone. In sympatry, L. austrodigitalis was more abundant higher in the intertidal on rocks, whereas L. digitalis was more abundant lower in the intertidal on barnacles. This finding supports earlier claims of microhabitat partitioning in this sibling species pair. In addition to this finding, the transition zone between the species was found to have shifted substantially northward in only two decades, from Monterey Peninsula, CA to near Pigeon Point, CA, where L. digitalis previously dominated.     

Microsatellite data reveal fine genetic structure in male Guiana dolphins (<Emphasis Type="Italic">Sotalia guianesis)</Emphasis> in two geographically close embayments at south-eastern coast of Brazil

A large macrogeographic differentiation has been observed among Sotalia guianensis populations along the South American coast. However, no genetic structure has been detected so far in closely distributed populations of this species, even though it has been observed in other cetaceans. Here, we examined the fine scale population structure for the largest populations of S. guianensis inhabiting Sepetiba and Paraty embayments at the south-eastern coast of Rio de Janeiro, Brazil. Analysis of mitochondrial DNA (mtDNA) control region sequences failed to detect variability among sequences. Conversely, evidence of significant male population structure was found on the basis of ten nuclear microsatellite loci. Surprisingly, the microsatellite markers were able to distinguish between individuals from the two embayments located 60 km apart. The results suggest that differences in habitat type and behavioral specializations are likely to explain the patterns of genetic structure. These findings should provide baselines for the management of communities exposed to increasing human-driven habitat loss.     

Genetic structure of populations of two species of Chthamalus (Crustacea: Cirripedia) in the north-east Atlantic and Mediterranean

Protein electrophoresis on starch gels was used to investigate population genetic structure of the barnacles Chthamalus montagui Southward and C. stellatus (Poli) over their north-east Atlantic and Mediterranean ranges. In each species, a single locus exhibited marked differentiation of allele frequencies between Atlantic and Mediterranean localities; in C. stellatus, genetic differentiation between the two basins had not previously been noted. In both species, mean heterozygosity per locus appeared higher in the Mediterranean samples than in the Atlantic, and Mediterranean populations had more alleles at the loci studied. Possible explanations for the differentiation between the Atlantic and the Mediterranean populations are discussed. Received: 30 May 1996 / Accepted: 17 September 1996     

Genetic differentiation in two sibling species of the brackish-water polychaete Hediste japonica complex (Nereididae)

Genetic divergence among ten populations of small- and large-egg forms of the brackish-water polychaete Hediste japonica complex was investigated on 14 isozyme loci by electrophoretic analysis. The two forms were distinguishable by complete allele substitutions at five loci, resulting in high genetic differentiation (Nei's D: 0.533 to 0.662). No genetic evidence of hybridization between the two forms was detected in sympatric populations in three rivers. These results indicate that the two forms are reproductively isolated, clearly showing that the two forms are distinct species. The genetic differentiation among populations was higher in the large-egg form (D: 0.005 to 0.111, G _ST: 0.435) than that in the small-egg form (D: 0.000 to 0.001, G _ST: 0.020). This genetic difference between the two forms seems to be attributable to a difference in their life histories. The average expected heterozygosity was low in populations of both the large-egg form (0.005 to 0.068) and the small-egg form (0.014 to 0.038) in comparison with other marine invertebrates. Received: 11 April 1997 / Accepted: 8 September 1997     

Allozyme electrophoretic analysis of the Hediste limnicola — H. diversicolor — H. japonica species complex (Polychaeta: Nereididae)

Three morphologically similar species of the polychaete genus Hediste [H. limnicola (Johnson, 1901), H. diversicolor (O. F. Müller, 1776), and H. japonica (Izuka, 1908)] have been described, respectively, from the west coast of North America, Europe, and Japan. Although these taxa are geographically isolated by oceans, the morphological criteria by which they can be separated are sufficiently variable as to make identification difficult. We have compared these three taxa biochemically using ten allozyme loci and have found that they are genetically distinct and constitute valid species. H. japonica (collected in June 1990 from Kyushu, Japan) appears more closely related to H. limnicola (collected in July 1989 from two sites in California and two in Oregon, USA) than it is to H. diversicolor (collected in October-November 1989 from the Weser estuary and the Jadebusen in Germany). H. limnicola has been recognized previously as a self-fertilizing hermaphrodite, but it is quite polymorphic in the four populations examined and we suggest that some cross-fertilization must occur in the field. Marked allele frequency differences between the two H. diversicolor populations examined, which were separated by only about 30 km, suggested that there was restricted gene flow between populations, or that there had been strong short-term selection.     

Spatial variation in dimethylsulfoniopropionate (DMSP) production in <Emphasis Type="Italic">Ulva lactuca</Emphasis> (Chlorophyta) from the Northeast Pacific

Although dimethylsulfoniopropionate (DMSP) has a variety of functions in marine macroalgae including that of a cryoprotectant, an osmolyte, a way to remove excess sulfur and energy, an antioxidant, and an allelopathic precursor, the latter two functions are believed to be the most important in Ulva lactuca L. (=U. fenestrata) in intertidal populations on the coast of Washington state, USA. The present study found significant variation in DMSP concentrations among U. lactuca collected in May 2005 from six sites ranging from 47°54.45′N (Possession Point, Whidbey Island, WA, USA) to 48°30.55′N (Shannon Point Beach, Anacortes, WA, USA), and also among individuals within sites, and among tissues (basal tissues near the holdfast, middle of the blades, and tips). Concentrations ranged from 37 to 224 μmol g⁻¹ fresh mass (FM). In several 10-day experiments between July 2001 and August 2004 with U. lactuca collected from several places on the coast of Washington, the effects of nutrient level (DIN), light intensity and wavelength, and grazing by the herbivorous gastropod Lacuna vincta, were examined. None of these manipulations resulted in DMSP concentrations that differed significantly from controls, and variance in DMSP concentrations within each experiment was very low. Although DMSP concentrations in U. lactuca may be affected by factors not tested in these experiments, it is also possible that the observed spatial differences reflect constitutive genotypic or phenotypic differences among geographically separated U. lactuca populations or among cryptic Ulva species. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Mitochondrial DNA variation and population genetic structure of the blue crab<Emphasis Type="Italic"> Callinectes sapidus</Emphasis> in the eastern United States

The genetic population structure of red grouper, Epinephelus morio (Valenciennes), and scamp, Mycteroperca phenax Jordan and Swain, from the southeastern U.S. Atlantic coast and the Gulf of Mexico was examined using nuclear microsatellite DNA markers in order to test the null hypothesis of panmixia throughout this range. Physical and biological data indicate that relatively isolated populations of these fish exist. Genetic variation was assessed at four microsatellite loci in red grouper and six loci in scamp. The fish were collected on different dates between 1991 and 2001. The microsatellite loci were highly polymorphic, with an average expected heterozygosity of 0.75 in red grouper and 0.68 in scamp. Heterozygote deficiencies (significant deviations from Hardy–Weinberg equilibrium, HWE) were found at two of four loci in all red grouper samples except the eastern Gulf of Mexico, and for all red grouper combined. In contrast, all loci conformed to HWE in the separate scamp samples. Minimal genetic differences distinguished southeastern U.S. Atlantic or Mexican red grouper from other localities, and no indication of genetic differentiation was observed in scamp. This large-scale genetic homogeneity may be attributed to ongoing gene flow and/or historical contact between present-day populations. For management purposes, genetic homogeneity does not necessarily imply a single stock. Because larval dispersal may be sufficient to homogenize gene frequencies but not to replenish depleted stocks, other data must be considered in the management of these species.Electronic Supplementary Material Supplementary material is available in the online version of this article at .Communicated by J.P. Grassle, New Brunswick     

Genetic divergence and phylogenetic inferences in five species of Mugilidae (Pisces: Perciformes)

Allozyme electrophoresis was used to compare the genetic divergence of global populations of Mugil cephalus Linnaeus, 1758 and two congeneric [M.␣curema Cuvier & Valenciennes, 1836; M. gyrans (Jordan & Gilbert, 1885)] and two more distantly related [Liza ramada (Risso, 1826); Xenomugil thoburni (Jordan & Starks, 1896)] species on the basis of 18 enzyme loci. The amount of genetic divergence among the species examined is in agreement with their present systematic status, the differences being larger among non-congeneric species than among species of the same genus. Intraspecific genetic distances in M. cephalus (average Nei's D= 0.154), although higher than those reported for conspecific populations of fish, appear to be small when compared to the interspecific values among mugilid species (0.821 ≤ Nei's D ≤ 1.744). Phylogenetic trees obtained by genetic distance methods and discrete character parsimony analysis were of similar topology, except for the relationships within the genus Mugil and for the arrangement of M. cephalus populations. Received: 7 April 1997 / Accepted: 4 February 1998     

Mode of larval development and genetic population structure inNodilittorina africana knysnaensis (Prosobranchia: Littorinidae)

We examined electrophoretic variation in the supralittoral gastropod,Nodilittorina africana knysnaensis, along the coast of South Africa in 1989. There was little allozyme variation in nine samples collected over 1 500 km for three highly polymorphic loci,Est, Gpi, andPgm, and Wahlund's effect was absent in the pooled sample. A gene diversity analysis showed that 98.1% of total genic diversity, on average, was contained within populations, and that only 1.9% was due to allozyme frequency differences among populations. Since the mode of larval development has not yet been described for this species, we suggest that the lack of strong allozyme frequency differences over long distances indicates thatN. a. knysnaensis has a planktonic mode of larval development.     

Meiofaunal cryptic species revealed by confocal microscopy: the case of Xenotrichula intermedia (Gastrotricha)

The phylum Gastrotricha includes about 750 species of meiobenthic marine and freshwater species that are often widely distributed. The microscopic size, short life cycle, low motility of adults, and the absence of larval stages normally required for dispersal raise doubts about the putative cosmopolitan distribution of many of gastrotrich species. The phenomenon of cosmopolitanism is acknowledged for all major meiobenthic taxa (“the meiofauna paradox”) and can be explained, at least in part, with the existence of sibling species, so far identified primarily by molecular analysis. In this paper, we report the discovery of sibling species in the marine chaetonotidan Xenotrichula intermedia using confocal laser scanning microscopy (CLSM). A total of 40 specimens collected from two geographically separate populations, the Mediterranean (Adriatic Sea) and the Arabian Gulf (Kuwait), were investigated. Fifteen specimens of each population were studied in vivo with a contrast interference microscope (DIC) in order to obtain the main morphometric parameters; ten other animals (five in each population) were fixed and marked with fluorescent phalloidin for the observation of their muscular systems under CLSM. The metrics and meristic data of the two populations fall within the range of measures recognized for the species. Pairwise comparisons (t-test) in general did not reveal statistically significant differences between the traits of specimens belonging to the two populations; moreover, multivariate analyses (cluster- and MDS analysis) were unable to separate clearly the Mediterranean from the Arabian specimens. In contrast, an examination of their muscular systems revealed clear dissimilarities between the two geographic groups. In particular, while the Italian specimens possessed incomplete circular bands and dorsoventrally orientated muscles that are partially inserted into the basal lamina of the cuticle, in both splanchnic- and somatic positions, their Arabian counterparts showed partial dorsoventral bands and complete circular muscles that surround the entire body of the animal in the same corresponding positions. Additional differences in the position of helicoidal bands (present in a more anterior location in the Italian specimens) are noted between Mediterranean and Arabian specimens. Since the investigated animals share a similar interstitial habitat, a working hypothesis connects the differences in the musculature to possible differences in the reproductive modality of specimens between the two populations.     

Microsatellite DNA markers and morphometrics reveal a complex population structure in a merobenthic octopus species (<Emphasis Type="Italic">Octopus maorum</Emphasis>) in south-east Australia and New Zealand

Five polymorphic microsatellite loci were developed and then used to assess the population genetic structure of a commercially harvested merobenthic octopus species (Octopus maorum) in south-east Australian and New Zealand (NZ) waters. Beak and stylet morphometrics were also used to assess population differentiation in conjunction with the genetic data. Genetic variation across all loci and all sampled populations was very high (mean number alleles = 15, mean expected heterozygosity = 0.85). Microsatellites revealed significant genetic structuring (overall F _ST = 0.024, p < 0.001), which did not fit an isolation-by-distance model of population differentiation. Divergence was observed between Australian and NZ populations, between South Australia and north-east Tasmania, and between two relatively proximate Tasmanian sites. South Australian and southern Tasmanian populations were genetically homogeneous, indicating a level of connectivity on a scale of 1,500 km. Morphometric data also indicated significant differences between Australian and NZ populations. The patterns of population structuring identified can be explained largely in relation to regional oceanographic features.