Cryptic speciation in a high gene flow scenario in the oviparous marine sponge Chondrosia reniformis

Sponge systematics has been traditionally based on the study of the skeleton (spicules and spongin fibres). However, sponges of the genus Chondrosia are devoid of those skeletal features, making it difficult to distinguish between different species in the genus. Chondrosia reniformis Nardo, 1847, the type species of the genus, was described from the Mediterranean Sea. The lack of distinguishing morphological features may have been responsible for the widespread assignment of specimens of the genus to this species; as a result C. reniformis is considered to be a cosmopolitan species. In this work, populations of C. reniformis from the western Mediterranean (France) and the West Atlantic (Bermuda and Brazil) were analysed using allozyme electrophoresis for 13 enzyme loci. Levels of mean heterozygosity were high (Bermuda and Brazil H=0.27 and W Mediterranean H=0.12), as is often observed in sponge species. Gene identities observed between West Atlantic and Mediterranean populations were low (I=0.40-0.52, typical values for congeneric species), including the presence of four diagnostic loci. This level of divergence clearly shows that they are not conspecific. Hence, a worldwide or cosmopolitan distribution of C. reniformis would seem improbable. However, the West Atlantic samples (Bermuda and Brazil) were genetically similar (gene identity, I=0.88-0.95) over a distance of 8,000 km. This is the first report of genetic homogeneity in a sponge species over such a large geographical distance.     

Global phylogeography of<Emphasis Type="Italic"> Cassiopea</Emphasis> (Scyphozoa: Rhizostomeae): molecular evidence for cryptic species and multiple invasions of the Hawaiian Islands

The upside-down jellyfish Cassiopea is a globally distributed, semi-sessile, planktonically dispersed scyphomedusa. Cassiopea occurs in shallow, tropical inshore marine waters on sandy mudflats and is generally associated with mangrove-dominated habitats. Controversy over the taxonomy of upside-down jellyfishes precedes their introduction to the Hawaiian Islands during the Second World War, and persists today. Here we address the global phylogeography and molecular systematics of the three currently recognized species: Cassiopea andromeda, C. frondosa, and C. xamachana. Mitochondrial cytochrome c oxidase I (COI) sequences from Australia, Bermuda, Fiji, the Florida Keys, the Hawaiian Islands, Indonesia, Palau, Panama, Papua New Guinea, and the Red Sea were analyzed. Highly divergent COI haplotypes within the putative species C. andromeda (23.4% Kimura 2-parameter molecular divergence), and shared haplotypes among populations of two separate putative species, C. andromeda and C. xamachana from different ocean basins, suggest multiple anthropogenic introductions and systematic confusion. Two deeply divergent Oahu haplotypes (20.3%) from morphologically similar, geographically separate invasive populations indicate long-term (14–40 million years ago) reproductive isolation of phylogenetically distinct source populations and cryptic species. Data support at least two independent introductions to the Hawaiian Islands, one from the Indo-Pacific, another from the western Atlantic/Red Sea. Molecular phylogenetic results support six species: (1) C. frondosa, western Atlantic (2) C. andromeda, Red Sea/western Atlantic/Hawaiian Islands (3) C. ornata, Indonesia/Palau/Fiji (4) Cassiopea sp. 1, eastern Australia (5) Cassiopea sp. 2, Papua New Guinea and (6) Cassiopea sp. 3, Papua New Guinea/Hawaiian Islands.Communicated by P.W. Sammarco, Chauvin     

Genetic divergence between east and west Atlantic populations of<Emphasis Type="Italic"> Actinia</Emphasis> spp. sea anemones (Cnidaria: Actiniidae)

The sea anemone Actinia equina was considered a highly variable species with a wide geographical distribution, but molecular systematic studies have shown that this wide distribution may be the result of the lumping of cryptic species. In this work enzyme electrophoresis was used to analyse the genetic variability of A. equina from the Atlantic coasts of Europe and Africa, as well as the relationships between those populations and other species of the genus. Samples of A. equina from the United Kingdom and France were compared with supposedly conspecific populations from South Africa and a recently described species from Madeira, Actinia nigropunctata. The South African and Madeiran populations were genetically very divergent from each other (genetic identity, I=0.15), as well as from the A. equina population from the United Kingdom and from the other species studied (I between 0.23 and 0.78), indicating that the African population of A. equina belongs to a different species. High genetic similarities were found between west Atlantic A. bermudensis and east Atlantic A. sali, in spite of the great geographical distance between them, and between the European A. equina and A. prasina. The results confirm the presence of the new species A. nigropunctata from Madeira and suggest a new species within the A. equina species group, from South Africa.Communicated by O. Kinne, Oldendorf/Luhe     

Spatial and behavioral interactions between a native and introduced salamander species

Behavioral interactions with native species may influence the invasiveness of introduced species. The salamanders Plethodon glutinosus and P. jordani in the eastern United States share many life history traits and demonstrate complex interspecific interactions that range geographically from competitive exclusion to sympatry. P. jordani was introduced to Mountain Lake Biological Station, Virginia, USA, between the years 1935 and 1945. We tested whether competition for space may influence the invasion of P. jordani into native P. glutinosus habitat by utilizing data from natural distributions, a field experiment, and controlled laboratory experiments. No environmental variables differed where P. glutinosus and P. jordani were collected in the field at the site of P. jordani introduction. In the field experiment, P. glutinosus was more fully exposed during foraging bouts in cages shared with heterospecifics as opposed to ones shared with conspecific salamanders. Condition (mass relative to body length) of salamanders at the end of the 3 months did not differ between conspecific and heterospecific treatments. In the laboratory, P. glutinosus most often attained the single burrow in the arena, but residency status had no effect. Species cohabited the burrow 50% of the time. Pair-wise encounters in the laboratory indicated that both species spend less than 20% of the time in aggressive behaviors as juveniles. Adults showed no behavior interpreted by us as aggression during pair-wise encounters. Received: 19 December 1999 / Accepted: 18 March 2000     

Genetic structure and distribution ofPhotololigo spp. in Australia

Four species ofPhotololigo were identified using allozymes from the northern waters of Australia between the North West Shelf (114°00E) and Brisbane (153°00E). Two of these species fitted the gross morphology ofP. edulis documented from Japan and two taxa shared the distinguishing features ofP. chinensis. F-statistic analyses revealed no major population genetic structuring in any of the species over the geographical range sampled and all populations were in Hardy-Weinberg equilibrium. Given that the distributions of the species appear to be correlated with depth, at least during the austral summer, this panmixia is considered to reflect extensive gene flow through longshore movement of these squid. It is suggested that depth constraints, or factors associated with depth, act as effective barriers to gene flow and, therefore, provide mechanisms for allopatric speciation in this genus. The data from northern Australia indicate that the current view thatLoligo species have broad distributions may need revision and that a number of widespread taxa (such asP. edulis andP. chinensis) are likely to comprise a series of hitherto unrecognised allopatric sibling species. Heterozygosity levels were low for all species exceptPhotololigo sp. 1, which had a restricted deep-water distribution. These data are concordant with the habitat specialist-generalist model, which is discussed.Contribution No. 654 from the Australian Institute of Marine Science     

Influence of sponges on invertebrate recruitment: A field test of allelopathy

We manipulated live sponges in Belize, Central America, Big Pine Key, Florida (USA), and Indian River lagoon, Florida (USA) in summer/autumn, 1988. At each location, live sponges of three species were placed within 0.5 cm of ceramic tiles. Tiles with synthetic sponges positioned in the same manner and tiles with no sponges served as controls. Of 26 recruiting species analyzed, only one (Sponge sp. 6 — Indian River) was inhibited by living sponges. Four species (Perophora regina — Belize;Aiptasia pallida — Big Pine Key; andCrassostrea virginica andAscidia nigra — Indian River) recruited in greater numbers in the presence of sponges, suggesting that some larvae may be attracted rather than repelled by sponge allelochemicals. Allelopathic effects were less important than small-scale flow effects and patchy larval supply in determining recruitment patterns on surfaces adjacent to sponges.     

Population differentiation and taxonomic status of the exploited limpet Patella candei in the Macaronesian islands (Azores,Madeira, Canaries)

There has been considerable confusion in the taxonomy of limpets of the North East Atlantic Ocean and Mediterranean Sea, particularly those from the Macaronesian islands. The present study compared populations of the intertidal limpet Patella candei d'Orbigny from the Azores, Madeira and the Canaries with those of P. caerulea Linnaeus and P. depressa Pennant from the European and African continental coasts. No major differences in radular morphology were detected between the three species. However, electrophoretic analysis of 15 enzyme loci gave overall genetic identity (I) values of 0.5 between the three species, indicating that they cannot be regarded as conspecific as previously thought, and suggesting that P. candei is endemic to the Macaronesian islands. Comparisons of P. candei within these islands showed that, although populations did not differ with respect to radular morphology and soft-body parts, populations from the Azores were distinct from those in Madeira and the Canaries in shell shape and gene frequencies. Individuals from the Azores had, no average, taller shells and longer radulae, while those in Madeira and the Canaries had a shallow, depressed and stellate shell form. This was interpreted as being due to the wider habitat distribution of the species in the Azores compared to Madeira and the Canaries. Electrophoretic results showed that P. candei from the Azores differed from P. candei in Madeira and the Canaries by almost 40% of the loci investigated (I=0.660), suggesting that the former is a separate endemic species. An I value of 0.969 between populations in Madeira and the Canaries was typical of conspecific populations.     

Population divergence in the sinistral whelks of North America,with special reference to the east Florida ecotone

This study evaluated models of species relationships among sinistral whelks in the genus Busycon in the Atlantic and Gulf of Mexico. Gene frequencies at eight polymorphic allozyme loci, shell morphology, anatomy, and partial DNA sequences for the cytochrome c oxidase I (COI) mitochondrial gene were examined in eight populations, ranging from New Jersey to the Yucatan peninsula, and from the dextrally coiled sister taxon Busycon carica (Gmelin, 1791). Whelks were collected in 1997 and 1998. The maximum COI sequence divergence recorded among 32 sinistral individuals was 1.96%, which together with the absence of any gross or qualitative morphological differences, suggested all eight populations should be considered conspecific. High levels of divergence between the allopatric western Atlantic and Gulf of Mexico populations, as revealed by fixed or nearly fixed differences at several allozyme-encoding loci were interpreted as evidence that the east Florida ecotone constitutes a significant barrier to gene flow. Size trimming also revealed several significant quantitative differences in shell and radular morphology between the three pooled Atlantic populations and five pooled Gulf populations. The Yucatan sample was the most distinctive conchologically, with heavy spines and tumid ridges, possibly related to stone crab predation. Based on the evidence all left-handed whelks of North America should be referred to the oldest available nomen, Busycon perversum (Linné, 1758), with three subspecies, B. perversum perversum along the Yucatan peninsula, B. perversum sinistrum (Hollister, 1958) in the northern and eastern Gulf of Mexico, and B. perversum laeostomum (Kent, 1982) in the Atlantic.Communicated by J.P. Grassle, New Brunswick     

Colonization of Europe by two American genetic types or species of the genus Marenzelleria (Polychaeta: Spionidae). An electrophoretic analysis of allozymes

Allozyme electrophoresis was conducted in an attempt to identify the origin ofMarenzelleria sp. found in the North Sea and Baltic Sea. The analysis covered eight enzymes with ten loti from nine populations found on the North American Atlantic toast, these populations in the North Sea and five populations in the Baltic. The North Sea spionids correspond to the Type IMarenzelleria from North American coastal waters between Barnstable Harbor (Massachusetts) and Cape Henlopen (Delaware). Nei's genetic distance between these North American populations and those from the North Sea wasD = 0.010 to 0.020. TheMarenzelleria sp. found in the Baltic Sea very probably stems from North American populations of Type II found from the region of Chesapeake Bay (Trippe Bay) south to the Currituck Sound (North Carolina). The genetic distance between these North American populations and the Baltic populations isD = 0.000 to 0.001. The invaders appear to have lost little of their genetic variation while colonizing the North and Baltic Seas. Probably, both colonizing events tan be attributed to large numbers of individuals reaching Europe simultaneously on one or more occasions. In addition, aMarenzelleria Type III was found by electrophoresis among specimens from Currituck Sound (North Carolina), rohere it is sympatric withMarenzelleria Type II. Salinity is discussed as an important factor for the establishment ofMarenzelleria Type I in the North Sea and Type II in the Baltic Sea.     

Distribution of overwintering calanoid copepod eggs in sea-bed sediments around southern Britain

Eggs of the calanoid copepodsLabidocera wollastoni (Lubbock) andCentropages hamatus (Lilljeborg) and others, which could not be reliably assigned to species, were identified from sediment samples taken during three cruises in December 1984, January 1986, and February 1987 in the southern North Sea, the English Channel, the Irish Sea and the Celtic Sea. Eggs were most abundant in sediment samples taken between 20 and 80 m depth, with bottom stress less than 10 dyn cm^–2, and near to the positions of tidal fronts. It is suggested that sedimentation, resuspension and sediment transport contribute to the causes of these distributions, which have been found to be consistent with the distributions of species which are known to produce diapause eggs as an overwintering strategy. Nauplii ofL. wollastoni, C. hamatus, C. typicus (Krøyer)Temora longicornis (Müller) andAcartia sp. hatched from incubated sediment samples. The last noted hatchings ofL. wollastoni, Centropages spp. andT. longicornis all occurred more than 1 yr after the start of the incubation experiments.     

Are meiofaunal species cosmopolitan? Morphological and molecular analysis of Xenotrichula intermedia (Gastrotricha: Chaetonotida)

Many meiofaunal species are reported to be cosmopolitan, but due to uncertainties of identification, the affiliation of specimens from geographically distant areas to the same species-taxon is problematic. In this study, we examined morphological and molecular variation in samples of Xenotrichula intermedia Remane (Gastrotricha: Chaetonotida) from the Mediterranean Sea, the northwestern Atlantic and the northern Gulf of Mexico. Univariate analysis of 16 morphological traits was unable to detect differences among populations, except for the length of the pharynx, which was significantly shorter in the Gulf of Mexico specimens. Canonical discriminant analysis separated the Gulf of Mexico specimens from the other two populations, with pharynx length contributing about half of the total discrimination. Molecular analysis based on restriction-fragment length polymorphisms (RFLPs) in a 710-base pair polymerase chain-reaction (PCR) produet representing roughly half of the mitochondrial cytochrome oxidase I (COI) gene detected four haplotypes: one each from the Mediterranean and the Gulf of Mexico populations and two coexisting within the Atlantic population. The estimated nucleotide-sequence divergence calculated for each pairwise combination of haplotypes (based on the proportion of shared fragments) ranged from 5.3 to 11.5%. The high genetic divergence and the inability to clearly separate populations based on morphology suggest that individuals characterized by different haplotypes are genetically isolated sibling species.     

Genetic structure of populations of two closely related brittle stars with contrasting sexual and asexual life histories,with observations on the genetic structure of a second asexual species

Ophiocoma pumila Lütken andOphiocomella ophiactoides (H. L. Clark) are morphologically similar brittle stars with contrasting life histories, the former obligately sexual, the latter fissiparous (capable of both sexual reproduction and asexual proliferation by binary fission). Electrophoretic analysis of five polymorphic enzymes was used to assess the genetic consequences of these differing life histories and provide a genetic perspective on the taxonomic relationship between the two species. Genotypic diversity ofOphiocoma pumila collected at Discovery Bay, Jamaica, in 1985 conformed to expectations for a sexually reproducing population. In contrast, genotypic diversity ofOphiocomella ophiactoides at this site was significantly lower than expected for a sexually reproducing population, due largely to the predominance of clonal proliferation over larval recruitment. Large variation in clonal composition over a short (50 m) distance emphasized the very localized scale of clonal mixing in this species. Allozymic data are indicative of a close sibling species relationship betweenOphiocoma pumila andOphiocomella ophiactoides which suggests that the present generic separation of the two species should be re-examined. Electrophoretic analysis was also used to examine the genetic structure of sponge- and alga-dwelling populations of a second fissiparous brittle star,Ophiactis savignyi (Müller & Troschel), which was also collected at Jamaica in 1985. Striking differences in the allelic composition of sponge- and alga-dwellingO. savignyi were observed. Genotypic diversity ofO. savignyi in sponges was very low, each sponge being dominated by a single genotype. Genotypic diversity ofO. savignyi in algae was higher, although still significantly lower than expectations for a sexually reproducing population. In the light of the highly clonal composition of fissiparous brittle-star populations, the adaptive significance of clonal growth may be related to an increase in the overall fitness of dispersed clones (genets), compared to individuals of strictly sexual counterparts, through greater genotypespecific biomass and, hence, fecundity.Contribution No. 476 of the Discovery Bay Marine Laboratory of the University of the West Indies     

Video-monitored predation by Caribbean reef fishes on an array of mangrove and reef sponges

Although predation by fishes is thought to structure benthic invertebrate communities on coral reefs, evidence to support this claim has been difficult to obtain. We deployed an array of eight sponge species on Conch Reef (16 m depth) off Key Largo, Florida, USA, and used a remote video-camera to record fish activity near the array continuously during five daylight periods (6 h for 1 d, at least 11.5 h for 4 d) and one night period (11 h). Of the eight sponge species, four were from adjacent reefs (Agelas wiedenmayeri, Geodia neptuni, Aplysina fistularis, and Pseudaxinella lunaecharta), and four were from a nearby mangrove habitat (Chondrosia collectrix, Geodia gibberosa, Halichondria sp., andTedania ignis). Each species of reef sponge was chosen to match the corresponding mangrove species in form and color (black, brown, yellow, and red, respectively). Predation events only occurred during daylight hours. Tallies of the number of times fishes bit sponges revealed intense feeding by the expected species of sponge-eating fishes, such as the angelfishHolacanthus bermudensis, H. tricolor, andPomacanthus arcuatus, the cowfishLactophrys quadricornis, and the filefishCantherhines pullus, but surprisingly also by the parrotfishSparisoma aurofrenatum andS. chrysopterum. Of 35 301 bites recorded, 50.8% were taken by angelfish, 34.8% by parrotfish, and 13.7% by trunkfish and filefish. Mangrove sponges were preferred by all reef fishes; 96% of bites were taken from mangrove species, with angelfish preferringChondrosia collectrix and parrotfish preferringGeodia gibberosa. Fishes often bit the same sponge repetitively, and frequently consumed entire samples within 30 min of their deployment. Sponge color did not influence fish feeding. Two of the four mangrove sponge-species deployed on the array were also found living in cryptic habitats on adjacent reefs and were rapidly consumed by fishes when exposed. Our results demonstrate the importance of fish predation in controlling the distribution of sponges on Caribbean reefs.     

Pheromones inBlattella germanica andBlaberus craniifer (Blaberoidea): Glandular source,morphology and analyses of pheromonally released behaviours

Females ofBlattella germanica andBlaberus craniifer produce a volatile sex pheromone attractive at a distance for conspecific males. During the emission of the pheromone, females ofB. craniifer adopt a typical calling posture; we never observe such a stance inB. germanica females. For both species, the glandular structures responsible for pheromonal production are located on the female pygidium. InB. germanica, the thickness of the glandular epithelium is clearly correlated with the attractiveness of females, being maximum for 13 day old females. The functions of the pygidial glands are discussed in both species.     

Allozyme and morphological evidence for a newly introduced species of Aurelia in San Francisco Bay,California

The jellyfish Aurelia aurita (Linnaeus, 1758) is usually considered to be a cosmopolitan species. Aurelia sp. medusae observed at Foster City, San Francisco Bay, California, USA, since 1988 are morphologically distinct from Aurelia sp. collected 200 km away in Monterey Bay, but are morphologically similar to aquarium-cultured Aurelia sp. from Japan. They differ consistently in radial canal morphology. In allozyme electrophoresis, significant differences at 12 of 14 polymorphic loci strongly suggest that Aurelia sp. from Foster City and Tokyo Bay belong to one species, while Aurelia sp. from Monterey Bay and Vancouver Island belong to a second species. We propose that Aurelia sp. at Foster City is a recent introduction, possibly from Japan via ships' ballast water. The identities and taxonomic affinities of the two Aurelia defined in this study, and their relationships with the Linnaen A. aurita described from the North Atlantic, will require genetic and morphological study of the currently recognized species A. aurita and A. limbata (Brandt, 1838) from several zoogeographical provinces.     

Evolution of coral reef fish<Emphasis Type="Italic"> Thalassoma</Emphasis> spp. (Labridae). 2. Evolution of the eastern Atlantic species

The genetic relationships within and among congeneric species of marine fish from the Atlantic and the Mediterranean are poorly known. Relationships among all five species of the wrasse genus Thalassoma present in the Atlantic and the Mediterranean were examined using sequence data from the mitochondrial control region. Sampling was focused on the mid-Atlantic T. sanctaehelenae (Valenciennes, 1839) and T. ascensionis (Quoy & Gaimard, 1834), the eastern Atlantic T. newtoni (Osório, 1891) from Sao Tome, and the eastern Atlantic/Mediterranean T. pavo (Linnaeus, 1758). Two western Atlantic species T. bifasciatum (Bloch, 1791) from the Caribbean and T. noronhanum (Boulenger, 1890) from Brazil served as outgroups. Tissues from a total of 132 individuals were sequenced. T. newtoni from Sao Tome preferentially grouped with the central Atlantic T. sanctaehelenae and T. ascensionis. T. pavo exhibits two distinct coloration patterns, one in the Cape Verde Islands and one in the eastern Atlantic Islands and Mediterranean. However, no genetic discontinuities between the Cape Verde Islands and the remaining samples or between Atlantic and Mediterranean individuals were found. Within Mediterranean populations of T. pavo, our data suggested the presence of a genetic break between eastern and western regions.Communicated by J.P. Grassle, New Brunswick     

Late Pleistocene dispersal of Indian-Pacific sardine populations in an ancient lineage of the genusSardinops

Temperate sardines fall into two related monotypic genera,Sardina andSardinops. Sardina exists as a cluster of subpopulations in the northeastern Atlantic and Mediterranean, andSardinops encompasses five geographically-isolated regional populations: (1) South Africa-Namibia, (2) Australia-New Zealand, (3) Chile-Peru, (4) Mexico-California and (5) Japan-Russia. We surveyed electrophoretic variability in the products of 34 protein encoding loci inSardina (N=26) and the five Indian-Pacific populations ofSardinops (N=222), collected from 1983 to 1991. Nei's genetic distances () between samples ofSardina andSardinops averaged 1.04 and are typical of distances between species of related genera.s between the regional forms ofSardinops were <-0.011, indicating thatSardinops consists of a single species with widely-scattered subpopulations. Assuming a molecular clock calibrated by the rise of the Panama Isthmus and the opening of the Bering Strait, these genetic distances correspond to times since divergence of <200 000 yr. AlthoughSardinops populations showed a significant degree of allele-frequency heterogeneity (F _ST, a measure of population differentiation, averaged 0.085 over 8 polymorphic loci), the distribution of genetic distances and tests of allele-frequency heterogeneity could not distinguished between hypotheses of north-south antitropical or east-west oceanic dispersal. Low levels of gene diversity inSardinops and mutation-drift disequilibria are consistent with a strong reduction in population size before the Late Pleistocene dispersal to the corners of the Indian-Pacific Oceans of an ancestralSardinops population.     

Reproductive isolation among morphotypes of the Atlantic seastar species<Emphasis Type="Italic"> Zoroaster fulgens</Emphasis> (Asteroidea: Echinodermata)

Zoroaster fulgens is a slope-dwelling seastar species that is distributed throughout the Atlantic Ocean. Studies into the population structure and systematics of marine animals have increasingly found that species with a reported cosmopolitan distribution are, in fact, collections of closely related cryptic or sibling species. In the Porcupine Seabight, three morphotypes of Z. fulgens can be found that have a distribution that is stratified by depth. This study investigates the genetic divergence between these morphotypes using sections of the cytochrome oxidase 1 (COI) and 16S regions of the mitochondrial genome. Bathymetrically separated morphotypes of Z. fulgens are reproductively isolated over distances of approximately 1 km while gene flow occurs among morphotypes, along isobaths, over distances of approximately 900 km. Reproductive isolation on the continental slope may have occurred as a result of selection exerted by gradients of depth-correlated physical factors, such as pressure and temperature. However, allopatric speciation with subsequent range expansion may also explain the observed patterns of genetic divergence. Further investigation of radiation within this group may provide important information on the evolution of slope species. Taxonomic revision of the genus is required.Communicated by J.P. Thorpe, Port Erin     

<Emphasis Type="Italic">Patella aspera</Emphasis> and<Emphasis Type="Italic"> P. ulyssiponensis</Emphasis>: genetic evidence of speciation in the North-east Atlantic

The group of subspecies of Patella ulyssiponensis, described by Christiaens, was widely known as Patella aspera until recently. The group extends throughout the Mediterranean, on all Macaronesian islands, along the North African coast, and in Europe, as far north as southern Norway. Throughout its range it displays great variation in shell sculpture and colour. The aim of this work was to re-examine the various subspecies proposed by Christiaens and, genetically, to test the hypothesis that European continental populations belong to a different species than the one composed by Macaronesian (north-west African) populations. In the present work, this group was studied by allozyme electrophoresis of 21 loci and by six morphological variables. The monomorphic locus of malate dehydrogenase (Mdh-1) was found to be diagnostic for distinguishing European continental populations from those of north-west African archipelagos, confirming the specific status of both groups. The allele observed at this locus in the Macaronesian populations was novel, while European continental populations showed a plesiomorphic one, shared with all other north-west Atlantic patellids with the exception of Patina pellucida, suggesting a more recent origin of the Macaronesian species from the continental forms. Both species showed a genetic identity of 0.730±0.061, which allowed rough estimations of 6.5–3 Mya since speciation. The subspecies Patella ulyssiponensis deserta described by Christiaens was not confirmed by our genetic data. Other loci (glyceraldehyde-3-phosphate dehydrogenase and the second locus of lactate dehydrogenase) were partially diagnostic, in which both species showed different most common alleles. Morphologically, both species are easily recognised by shell characters and the results agree with previous findings, that continental populations are more homogeneous in shell morphology and radula characters than populations from the Macaronesia. This work supports retention of the earliest valid name, Patella ulyssiponensis Gmelin, with Lisbon, Portugal as type locality, for the European continental species, and Patella aspera Röding for the Macaronesian populations. Population subdivision within species was measured by theta, the estimator of F_st, showing in both P. aspera and P. ulyssiponensis a high degree of genetic structuring (=0.226 and 0.182, respectively) mostly explained by the large distances separating the populations within species.Communicated by J.P.Thorpe, Port Erin     

Cytotoxic haplosclerid sponges preferred: a field study on the diet of the dotted sea slug<Emphasis Type="Italic"> Peltodoris atromaculata</Emphasis> (Doridoidea: Nudibranchia)

Trophic specializations are widespread among opisthobranch molluscs. One purported example from the Mediterranean Sea is the dotted sea slug Peltodoris atromaculata. It has been hypothesized that this species is strongly monophagous on the sponge Petrosia ficiformis. However, the small amount of evidence that has been found for this hypothesis is based just on laboratory tests. Here we study the feeding habits and the diet of Peltodoris atromaculata in its natural habitat. We observed and videotaped 161 individuals together with the organisms on which they were found (their living substrata). Feeding scars were identified and videotaped as well. Individuals and their living substrata were sampled for further analysis in the laboratory. The composition of faeces of Peltodoris, especially undigested sponge spicules, was analyzed by light and scanning electron microscopy and compared to the composition of the living substrata. Most of the faecal samples consisted of undigested sponge spicules. Although Peltodoris was found on 11 species of sponges, only 2 of them, Petrosia spp. and Haliclona fulva, form its diet (76% out of n=121 samples). In accordance with this, feeding scars in the habitats were exclusively observed on these two sponges. Estimation of electivity indices suggests that Haliclona is preferred over Petrosia. One remarkable feature of the exclusive feeding of Peltodoris on Petrosia and Haliclona is that both sponges share specific fulvinol-like polyacetylenes that show cytotoxic activity in bioassays. Potential benefits and evolutionary aspects of this trophic specialization are discussed. Besides sponge-containing faeces, we found spicule-free faeces (24%, n= 29). These were very small in volume compared to sponge-containing faeces, and only few distinct structures were present. However, the use of food other than sponges is not necessarily indicated by this, because the spicule-free faeces might also represent left-overs from the stomach and digestive gland after sponge spicules have been released.Communicated by O. Kinne, Oldendorf/Luhe