Genetic structure of the flounders Platichthys flesus and P. stellatus at different geographic scales

The genetic structure of the flounders Platichthys flesus L. and P. stellatus Pallas was investigated on different spatial scales through analysis of allozyme variation at 7 to 24 polymorphic loci in samples collected from different regions (Baltic Sea, North Sea, Brittany, Portugal, western Mediterranean, Adriatic Sea, Aegean Sea and Japan) in 1984 to 1987. No geographic variation was evident within a region. Some pattern of differentiation by distance was inferred within the Atlantic, while the Mediterranean comprised three geographically isolated populations and was itself geographically isolated from the Atlantic (fixed allele differences at up to three loci were found among P. flesus populations from the Atlantic, the western Mediterranean, the Adriatic Sea, the Aegean Sea and also P. stellatus from the coast of Japan). Sea temperature during the reproductive period probably acts as a barrier to gene flow between populations. Genetic distances among European flounder populations (P. flesus) were higher than, or of the same magnitude as, the genetic distance between Pacific (P. stellatus) and European flounder populations, suggesting that P. flesus is paraphyletic and/or there is no phylogenetic basis to recognising P. stellatus as a different species. The divergence between P. flesus and P. stellatus was thus inferred to be more recent than the divergence between the present P. flesus populations from the NE Atlantic and eastern Mediterranean. The eastern Mediterranean populations are thought to originate from the colonisation of the Mediterranean by a proto-P. flesus/P. stellatus ancestor, whereas the present western Mediterranean population has undergone a more recent colonisation event by P. flesus. Patterns of mitochondrial DNA variation, established on a smaller array of P. flesus samples, were in accordance with the geographic patterns inferred from the allozyme survey. In addition, they supported the hypothesis of a two-step colonisation of the western Mediterranean. These results contribute to our understanding of the biogeography of the Mediterranean marine fauna, especially the group of boreal remnants to which P. flesus belongs. Received: 7 February 1997 / Accepted: 26 March 1997     

Mitochondrial DNA and electronic tracking reveal population structure of Atlantic bluefin tuna (Thunnus thynnus)

Population subdivision was examined in Atlantic bluefin tuna (Thunnus thynnus) through sequencing of the control region of the mitochondrial genome. A total of 178 samples from the spawning grounds in the Gulf of Mexico, Bahamas and Mediterranean Sea were analyzed. Among the samples from these locations were 36 electronically tagged bluefin tuna that were tagged in the North Atlantic and subsequently traveled to one of these known spawning grounds during the spawning season. Bluefin tuna populations from the Gulf of Mexico and the Mediterranean Sea were found to be genetically distinct based on Φ_st, and sequence nearest neighbor analyses, showing that these two major spawning areas support independent stocks. Sequence nearest neighbor analysis indicated significant population subdivision among the Gulf of Mexico, western Mediterranean and eastern Mediterranean Sea. However, it was not possible to find significant pairwise differences between any sampling areas when using all samples. If only samples that had a high likelihood of assignment to a specific spawning site were used (young of the year, spawning adults), the differentiation increased among all sampling areas and the Western Mediterranean Sea was distinct from the Eastern Mediterranean Sea and the Gulf of Mexico. It was not possible to distinguish samples from the Bahamas from those collected at any of the other sampling sites. These data support tagging results that suggested distinctness of the Gulf of Mexico, Eastern and Western Mediterranean Sea spawning areas. This level of stock differentiation is only possible if Atlantic bluefin tuna show strong natal homing to individual spawning grounds.     

Genetic diversity of blackspot seabream (<Emphasis Type="Italic">Pagellus bogaraveo</Emphasis>) populations off Spanish Coasts: a preliminary study

Genetic diversity among four natural samples of Blackspot seabream (Pagellus bogaraveo, Brünnich, 1768) from different fishing grounds exploited by Spanish fisheries was analyzed through the use of 12 microsatellite markers. The samples were captured off the Spanish coasts from the Mediterranean Sea to the Cantabrian Sea within the same continental slope. High levels of genetic diversity were revealed for every population and every locus was polymorphic at the 0.95 level. The average number of alleles, average heterozygosity and PIC were found to be 15.75, 0.833 and 0.818, respectively. In general, population differentiation was not detected in these samples. Through AMOVA, a low level of variation between regions (Mediterranean vs. Atlantic samples) was observed, though this was not significant. A larger percentage of total variation was observed inside the ‘within populations’ class. Thus, AMOVA did not reveal any significant population substructure. Moreover, no correlation was found between geographical and F_ST estimates and the observed results did not allow the improvement of a model of isolation by distance. The high homogeneity revealed by means of these markers could indicate the absence of physical frontiers between the geographical areas analyzed in this survey, especially between Atlantic and Mediterranean areas.     

Atlantic-Mediterranean and within-Mediterranean molecular variation in <Emphasis Type="Italic">Coris julis</Emphasis> (L. 1758) (Teleostei,Labridae)

Sequence variation in the mitochondrial control region was studied in the Mediterranean rainbow wrasse (Coris julis), a species with pronounced pelagic larval phase inhabiting the Mediterranean Sea and the adjacent coastal eastern Atlantic Ocean. A total of 309 specimens from 19 sampling sites were analysed with the aim of elucidating patterns of molecular variation between the Atlantic and the Mediterranean as well as within the Mediterranean Sea. Phylogeographic analyses revealed a pronounced structuring into a Mediterranean and an Atlantic group. Samples from a site at the Moroccan Mediterranean coast in the Alboran Sea showed intermediate frequencies of “Mediterranean” and “Atlantic” haplotypes. We recognised a departure from molecular neutrality and a star-like genealogy for samples from the Mediterranean Sea, which we propose to have happened due to a recent demographic expansion. The results are discussed in the light of previous studies on molecular variation in fish species between the Atlantic and the Mediterranean and within the Mediterranean.     

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     

Genetic analysis of tuna populations,<Emphasis Type="Italic"> Thunnus thynnus thynnus</Emphasis> and<Emphasis Type="Italic"> T</Emphasis>.<Emphasis Type="Italic"> alalunga</Emphasis>

The genetic population structures of Atlantic northern bluefin tuna ( Thunnus thynnus thynnus) and albacore ( T. alalunga) were examined using allozyme analysis. A total of 822 Atlantic northern bluefin tuna from 18 different samples (16 Mediterranean, 1 East Atlantic, 1 West Atlantic) and 188 albacore from 5 samples (4 Mediterranean, 1 East Atlantic) were surveyed for genetic variation in 37 loci. Polymorphism and heterozygosity reveal a moderate level of genetic variability, with only two highly polymorphic loci in both Atlantic northern bluefin tuna ( FH* and SOD- 1*) and albacore ( GPI- 3* and XDH*). The level of population differentiation found for Atlantic northern bluefin tuna and albacore fits the pattern that has generally been observed in tunas, with genetic differences on a broad rather than a more local scale. For Atlantic northern bluefin tuna, no spatial or temporal genetic heterogeneity was observed within the Mediterranean Sea or between the East Atlantic and Mediterranean, indicating the existence of a single genetic grouping on the eastern side of the Atlantic Ocean. Very limited genetic differentiation was found between West Atlantic and East Atlantic/Mediterranean northern bluefin tuna, mainly due to an inversion of SOD- 1* allele frequencies. Regarding albacore, no genetic heterogeneity was observed within the Mediterranean Sea or between Mediterranean and Azores samples, suggesting the existence of a single gene pool in this area.     

Population structure of the wreckfish Polyprion americanus determined with microsatellite genetic markers

 We examined population structure in the wreckfish, Polyprion americanus, by assaying six microsatellite loci in 422 individuals collected throughout the geographic range. Eighteen hapuku, P. oxygeneios, were assayed at the same loci for use as an outgroup. Expected heterozygosities ranged from 0.49 to 0.88 and averaged 0.66. Allele-frequency distributions at those loci indicated that samples from the eastern North Atlantic, western North Atlantic and the Mediterranean were genetically similar, confirming the pattern seen in a previous analysis of mitochondrial DNA (mtDNA) restriction fragment length polymorphisms (RFLPs). Both mtDNA and microsatellite studies differentiated northern and southern wreckfish stocks. However, in contrast to the mtDNA studies, allelic variation at microsatellite loci clearly differentiated wreckfish from two Southern Hemisphere locations, Brazil and the South Pacific. Far more genetic variation was observed at microsatellite loci than with mtDNA RFLPs (haplotype diversity averaged 0.01), and we saw more evidence of population structure with the microsatellite loci. The differentiation between southern and northern wreckfish supports the distribution records, which indicate that wreckfish do not occur in the tropics. Temperature profiles and current patterns throughout the southern oceans apparently also prevent significant gene flow between the South Pacific and Brazilian samples. Received: 29 January 2000 / Accepted: 27 June 2000     

Distribution of euphausiid assemblages in the Mediterranean Sea

Eleven species of euphausiids from 24 Isaac-Kidd Midwater Trawl (IKMT) night collections taken at stations throughout the Mediterranean Sea were counted. The frequency of occurrence and dominance of individual species and percent similarity faunal analysis of the euphausiid community were used to describe changes in faunal composition between geographical areas and differences in vertical distribution. Although most species were widespread, three distinct patterns of abundance were apparent: Euphausia krohnii, Nematoscelis megalops, Meganyctiphanes norvegica, and Stylocheiron abbreviatum predominated in western basin areas west of the Tyrrhenian Sea; Euphausia hemigibba, Thysanopoda aequalis, and Stylocheiron longicorne predominated in the Tyrrhenian Sea and eastern Mediterranean Sea; Euphausia brevis and Stylocheiron suhmii predominated in the eastern Mediterranean Sea. Percent similarity analysis of data from the IKMT samples and data from Ruud (1936) indicates the Tyrrhenian Sea fauna at the time of the collections was more similar to eastern Mediterranean areas than to most other areas in the western basin, although the degree of similarity was dependent, to some extent, on the depth at which the samples were collected. The composition of euphausiids living above 150 m at night in this area was more similar to eastern basin areas, while the composition of deeper living forms was more similar to those of the rest of the western basin. Comparison of euphausiids collected at three points over a 60 year time-span in the Balearic Sea shows the similarity in composition to be greater within the area over time than between adjacent areas in the western Mediterranean Sea.Contribution No. 2732 from the Woods Hole Oceanographic Institution, Woods Hole, Massachusetts, USA. This research was supported by the Atomic Energy Commission Contract AT (30-1)-3862, ref. NYO-3862-49, National Science Foundation Grant GA 29303 and Office of Naval Research Contract N00014-71-C-0284.     

Genetic structure of summer flounder (Paralichthys dentatus) populations north and south of Cape Hatteras

Genetic divergence among samples of summer flounder (Paralichthys dentatus, Linnaeus) was estimated to evaluate the effect of Cape Hatteras, a well-known zoogeographic barrier in the western Atlantic Ocean, on intraspecific gene flow. Previous studies based on meristic and morphometric variation and tagging data suggested that gene flow might be limited among summer flounder populations north and south of Cape Hatteras, although the high vagility of both juveniles and adults suggests otherwise. We analyzed genetic diversity revealed in the mitochondrial DNA (mtDNA) control region in samples of juveniles and adults collected in 1992 to 1996 from coastal sites from Buzzard's Bay, Massachusetts to Charleston, South Carolina. In contrast to previous morphological studies, analyses of mtDNA variation reveal no significant population subdivision centered around Cape Hatteras. Received: 17 September 1997 / Accepted: 8 September 1998     

Allozymes and morphometric characters of three species ofMytilus in the Northern and Southern Hemispheres

Many authors have considered the common mussels in temperate waters of the Northern and Southern Hemispheres to be a single cosmopolitan species,Mytilus edulis Linnaeus, 1758. Others have divided these mussels into several subspecies or species. Samples of mussels were collected from 36 locations in the Northern Hemisphere and nine locations in the Southern Hemisphere. Electrophoretic evidence from eight loci indicates that the Northern Hemisphere samples consist of three electrophoretically distinguishable species:M. edulis from eastern North America and western Europe;M. galloprovincialis Lamarck, 1819 from the Mediterranean Sea, western Europe, California, and eastern Asia; andM. trossulus Gould, 1850 from the Baltic Sea, eastern Canada, western North America and the Pacific coast of Siberia. Mussels from Chile, Argentina, the Falkland Islands and the Kerguelen Islands contain alleles characteristic of all three Northern Hemisphere species, but because they are most similar toM. edulis from the Northern Hemisphere, we suggest that they tentatively be included inM. edulis. These South American samples are morphologically intermediate between Northern HemisphereM. edulis andM. trossulus. Mussels from Australia and New Zealand are similar in allele frequency and morphometric characters toM. galloprovincialis from the Northern Hemisphere. FossilMytilus sp. are present in Australia, New Zealand and South America, which suggests that the Southern Hemisphere populations may be native, rather than introduced by humans. Morphometric characters were measured on samples which the allozyme data indicated contained a single species. Canonical variates analysis of the morphometric characters yields functions which distinguish among our samples of the species in the Northern Hemisphere.     

Genetic stock structure of the swordfish (Xiphias gladius ) inferred by PCR-RFLP analysis of the mitochondrial DNA control region

Restriction fragment length polymorphism (RFLP) analysis was performed on PCR amplified DNA fragments containing the control region of the swordfish (Xiphias gladius Linnaeus, 1758) mitochondrial DNA. A total of 456 individuals comprising 13 local samples (six Pacific, three Atlantic, the Mediterranean Sea, two Indian Ocean and the Cape of Good Hope) were surveyed with four endonucleases (Alu I, Dde I, Hha I and Rsa I), yielding a total of 52 composite genotypes. Within-sample genotypic diversity (H) was high ranging from 0.702 to 0.962 with a value of 0.922 for the pooled sample. Significant geographic variation in the frequencies of genotypes and restriction patterns was revealed. The Mediterranean sample was highly distinct from all other samples. Further, Rsa I digestion revealed high levels of polymorphism in all but the Mediterranean samples, indicating that exogenous swordfishes rarely enter that body of water. Heterogeneity between the North and South Atlantic samples was significant, both of which differed from those of the Pacific. In contrast, the Indian Ocean samples were not significantly different from the samples of South Atlantic and Pacific. Genetic differentiation among the Pacific samples was low. The results indicate that the world-wide swordfish population is genetically structured not only among, but also within ocean basins and suggest that gene flow is restricted despite the absence of geographic barriers. Received: 28 August 1996 / Accepted: 2 October 1996     

Optic gland enlargement and female gonad maturation in a population of the octopuseledone cirrhosa: a multivariate analysis

Eleven principal component analyses were performed on morphometric data from a series of monthly samples of the octopusEledone cirrhosa (Lamarck) collected from March 1978 through January 1979 in the Aberdeen (Scotland) area. This technique has allowed a partitioning of general growth effects from variation in the reproductive system. Optic gland size varies as a reproductive character together with ovary and oviducal glands. Optic gland size not clearly associated with other cerebral organs, the brain and optic lobes. The association between the reprountive characters developed during the onset of the main period of reproductive maturation in the early summer of 1978 and persisted for the remainder of that year. This result supports the experimental evidence linking optic gland enlargement with female gonad maturation and suggests that this process occurs normally in the field population.     

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 structuring of immature loggerhead sea turtles (Caretta caretta) in the Mediterranean Sea reflects water circulation patterns

The analysis of mitochondrial DNA in loggerhead sea turtles (Caretta caretta) from eight foraging grounds in the Mediterranean and the adjoining Atlantic revealed deep genetic structuring within the western Mediterranean. As a consequence, the foraging grounds off the North-African coast and the Gimnesies Islands are shown to be inhabited mainly by turtles of the Atlantic stocks, whereas the foraging grounds off the European shore of the western Mediterranean are shown to be inhabited mainly by turtles from the eastern Mediterranean rookeries. This structuring is explained by the pattern of sea surface currents and water masses and suggests that immature loggerhead sea turtles entering the western Mediterranean from the Atlantic and the eastern Mediterranean remain linked to particular water masses, with a limited exchange of turtles between water masses. As the north of the western Mediterranean comprises mostly individuals from the highly endangered eastern Mediterranean rookeries, conservation plans should make it a priority to reduce the mortality caused by incidental by-catch in these areas.     

Intraspecific geographic-morphological variation patterns in Fucus distichus and F. evanescens

The intraspecific patterns of morphometric variation were examined in Fucus distichus L. and F. evanescens C.Ag., using samples gathered from 49 sites spread across the North Atlantic. Ten characters were measured on each plant. The data were subjected to bivariate regression analyses, canonical correlation analysis and nested analysis of variance (ANOVA). It was shown that the combined effects of the three environmental variables measured for each site (latitude, longitude and exposure) could only explain 23 and 15% respectively of the intraspecific morphometric variation. Inter-individual, within-site differences accounted for a further 62 and 53%. The remaining variation appears to be expressed on a geographic scale of tens of kilometres at most. Thus, the species seem to take the form of fine-scale phenotypic mosaics, superimposed on weak major clines. Such a pattern has not previously been described for any macroalga, although it is known in some angiosperms.     

Mitochondrial DNA variation in king mackerel (Scomberomorus cavalla) from the western Atlantic Ocean and Gulf of Mexico

King mackerel (Scomberomorus cavalla Cuvier) collected in 1992 and 1993 from 13 localities along the Atlantic coast of the southeastern USA and in the northern Gulf of Mexico were surveyed for variation in mitochondrial (mt)DNA and a nuclear-encoded dipeptidase locus (PEPA-2). Both polymorphic and fixed mtDNA restriction sites were identified and mapped using conventional and polymerase chain-reaction (PCR)-based methods. Heterogeneity in mtDNA haplotype frequencies was found only in comparisons of pooled haplotypes from Atlantic localities versus pooled haplotypes from Gulf localities. This finding indicates weak genetic divergence between king mackerel from the Atlantic and those from the Gulf. Frequencies of two PEPA-2 alleles essentially paralleled previous findings: one allele (PEPA-2a) was common among samples from western Gulf localities, whereas the other allele (PEPA-2b) was common among samples from Atlantic and eastern Gulf localities. There was considerable variation in PEPA-2 allele frequencies within broadly-defined regions. Variation in mtDNA haplotypes and PEPA-2 genotypes was independent, as was variation in mtDNA haplotypes with sex or age of individuals. Variation in PEPA-2 genotypes was not independent of sex or age of individuals. The latter result suggests that frequencies of PEPA-2 alleles in samples of king mackerel may stem, in part, from sex and age distributions of individuals within samples, and indicates that caution should be exercised in using allelic variation at PEPA-2 as a measure of population (stock) structure in king mackerel. The discordance in spatial patterning of mtDNA haplotypes versus PEPA-2 alleles across the Gulf (i.e. homogeneity in mtDNA haplotype frequencies versus heterogeneity in PEPA-2 allele frequencies) may be due to either female excess at several localities, sex-biased migration, or both. Observed patterns of genetic variation also are consistent with the hypothesis that king mackerel in the western Atlantic may have been subdivided during Pleistocene glaciation, and that the current distribution of PEPA-2 alleles may be a historical artefact. Received: 17 December 1996 / Accepted: 2 April 1997     

Population structure of the black tiger prawn, Penaeus monodon, among western Indian Ocean and western Pacific populations

We examined the population structure of the black tiger prawn, Penaeus monodon Fabricius, 1798, in the Indo-West Pacific by analyzing the geographic distribution of elongation factor 1-alpha intron sequences from specimens collected during the winter and spring of 1997. Both the molecular phylogeny of alleles and F-statistics indicated very strong differentiation between populations from the western Indian Ocean and western Pacific. This pattern is concordant with other recent studies of marine species in this region, implying that the Indo-Australian Archipelago represents a biogeographic break between populations in the Indo-West Pacific. F _ST-values among populations in the western Indian Ocean also indicate structure within this region, whereas no structure was found among western Pacific populations. Nucleotide diversity was significantly lower in the western Indian Ocean populations than in the western Pacific, implying that the populations have regional differences in demographic history. Received: 16 November 1998 / Accepted: 26 May 1999     

Population differentiation in megrim (<Emphasis Type="Italic">Lepidorhombus whiffiagonis</Emphasis>) and four spotted megrim (<Emphasis Type="Italic">Lepidorhombus boscii</Emphasis>) across Atlantic and Mediterranean waters and implications for wild stock management

Megrim, Lepidorhombus whiffiagonis, and four spot megrim, Lepidorhombus boscii, are two marine fish species of high commercial interest. Despite their quite heavy exploitation little is known on the genetic structure of their populations. The present work aimed at characterizing the first seven microsatellites markers available for the two megrim species. These new markers were in a second step employed to describe the population structure of the two species among their almost entire habitat range (Atlantic and Mediterranean samples). Our study confirmed the existence of a strong genetic difference between Atlantic and Mediterranean megrim species already described in the literature for L. whiffiagonis on the basis of variations at ribosomal genes. Additionally our analysis gave the first evidences of a strong genetic differentiation among Atlantic populations in both megrim species (within Atlantic global F_ST in L. whiffiagonis and L. boscii were respectively 0.158 and 0.145). When describing megrim population structure, the comparison between allele-frequency-based tests (F_ST comparisons) and genotype-based inferences (Bayesian approach) gave evidences of a hierarchical structure of the populations. In conclusion, our work enlighten the existence of two different stocks within the Atlantic Ocean and one in the Mediterranean Sea that will clearly need to be managed separately. As the present results do not fully support the current megrim stock boundaries they will surely help to rethink megrim management policies in the future.     

Phylogeographic patterns of the mysid Mesopodopsis slabberi (Crustacea, Mysida) in Western Europe: evidence for high molecular diversity and cryptic speciation

The phylogeographic patterns among populations of Mesopodopsis slabberi (Crustacea, Mysida), an ecological important mysid species of marine and estuarine habitats, were analysed by means of DNA sequencing of a fragment of the mitochondrial cytochrome c oxidase subunit I (COI) and the 16S ribosomal RNA genes. Samples of M. slabberi collected from five Atlantic and two Western Mediterranean populations were investigated. Very high levels of within-population molecular diversity were observed in all samples (mean h=0.807 and π=0.0083), with exception of the Mediterranean Ebro population which contained only one haplotype. Differentiation among populations was high, and a clear phylogeographic break was observed between the Atlantic and Mediterranean populations. Moreover, a strong differentiation was detected between both populations in the Western Mediterranean Sea (Alicante and Ebro delta), while two divergent lineages occurred in sympatry within the Atlantic Mondego estuary. The high congruence between both the COI and 16S rRNA sequence data, the reciprocal monophyly of the different mitochondrial clades and the levels of nucleotide divergence between them suggest the presence of a complex of cryptic species within M. slabberi. Estimations of divergence time between the different mitochondrial lineages indicate that a split occurred during the late Miocene/early Pliocene. Such a divergence could be concordant with vicariant events during sea-level drops within the Mediterranean region at that time. However, within the Mediterranean Sea, the potential of divergence through ecological diversification cannot be ruled out.