A study of osteological and molecular differences in populations of Aphanius fasciatus Nardo 1827, from the central Mediterranean (Teleostei, Cyprinodontidae)

Nine populations of Aphanius fasciatus Nardo, 1827 from the central Mediterranean were analysed by examining the mitochondrial control region and the morphology of the bony elements of the skull and vertebral column, to study the degree of intraspecific differentiation of A. fasciatus considering the level of isolation of the different populations and the palaeogeographic history of the central Mediterranean area. Both the molecular and morphological analyses differentiate between the populations, even if the topologies of the two trees are different. Molecular and osteological investigations have consistently demonstrated a well-supported differentiation of the south-eastern Sicilian populations both within the same group (Tigano et al. in Ital J Zool 71:1124–1133, 2004a; Tigano et al. in Abstract volume XI European Congress of Ichthyology, Tallin, Estonia, 2004b), and from the populations from western Sicily, Tunisia and the island of Malta. The molecular results show that the nine populations are characterised by haplotypes that are well defined in relation to a probably limited gene flow; while, as regards the morphological data the differentiation found could be explained in terms of the geographic isolation of the various populations, although the influence of environmental factors, which differ greatly between the various sites where the populations live, cannot be ruled out.     

Phylogeography and genetic structure of the Mediterranean killifish <Emphasis Type="Italic">Aphanius fasciatus</Emphasis> (Cyprinodontidae)

The purpose of this study was to determine the phylogeographic structure of the brackish-hypersaline cyprinodont fish Aphanius fasciatus (Valenciennes, 1821), using sequencing and RFLP analysis of a 1,330 bp mitochondrial DNA segment containing part of the 16S rRNA gene as well as the genes for tRNA-Leu, NADH subunit 1 and tRNA-Ile. Individuals were collected from 13 different sites in Greece and Turkey, while seven published A. fasciatus sequences were also included to cover the area of distribution of the species. Pairwise sequence divergence values ranged from 0 to 4.51%. Congruent phylogenies were recovered with maximum likelihood, maximum parsimony and neighbour-joining methods. All analyses revealed two main groups. The first group consists of populations from almost all localities that drain into the Aegean Sea. The second group comprises the remaining population samples, which in some cases seem to consist of population-specific subgroups. Our results show that vicariant events have predominantly affected the evolution of A. fasciatus, with the Messinian salinity crisis having shaped the present genetic structure of its populations. Additionally, the life-history traits of the species, which determine a low potential for dispersal, coupled with the typical fragmentation of brackish-hypersaline water habitats have led to a high degree of isolation of A. fasciatus populations, even at restricted spatial scales. Analysis of the partitioning of the total amount of polymorphism with analyses of molecular variance (AMOVA) gave a value of F _ST = 84.6%. Potential conservation policies concerning A. fasciatus should also consider the low-genetic variability in the majority of its populations and the presence of fixed haplotypes in some of them.     

Molecular phylogeography of two Italian sibling species of Calobius (Coleoptera, Hydraenidae, Ochthebiinae) inhabiting Mediterranean marine rock-pools

Marine rock-pools, commonly found along the Mediterranean coasts, are isolated patches of habitat characterised by large spatial and temporal variations. The phylogeography of Calobius quadricollis and C. urbanelliae, two sibling species of moss beetles (Coleoptera, Hydraenidae) inhabiting Italian temporary marine rock-pool ecosystems, were studied using mitochondrial cytochrome c oxidase subunit I (COI). Our data suggest a strong association between the scored genetic variability and the geographical distribution of populations sampled for both species. These analyses provided evidences of episodic species range expansion and fragmentation, recurrent “flush and crash” and bottleneck episodes probably occurring during the Glacial Cycles. The observed phylogeographical pattern is probably related to the historical and biogeographical processes of Mediterranean areas, as well as both to the ephemeral habitat tipology and limited dispersal ability of these beetles.     

Social organization and sexual pattern in the Mediterranean parrotfish Sparisoma cretense (Teleostei: Scaridae)

We examined the social organization and reproductive pattern of a population of Sparisoma cretense L. at Lampedusa Island (Italy). During the breeding season (July to September) individuals occur either in territorial or in non-territorial groups, which quantitatively differ with respect to use of space, sex-ratio and sexual activity. Territorial groups consist of one male plus one to three females, whereas in non-territorial groups as many as 54 fish share a common area. Among non-territorial groups, variations, in both sex-ratio and behavior, suggest they could represent either feeding aggregations or reproductive aggregations. Spawning, always in pairs, occurs daily in a short period of time before dusk and has been observed only in territorial groups. Group spawning has never been observed, but another alternative mating tactic, streaking on pair spawning, was recorded. In the non-breeding season fish do not aggregate in organized social units. Histological examination of gonads showed that adults are larger than 12 cm total length. Females have an asynchronous ovary, typical of species spawning several times during the breeding season. Males show secondary testes, indicating that their gonads develop ovaries as juveniles, which are later redifferentiated into functional testes. Histological and demographic data seem to indicate that, as in other species of this genus, prematurational sex-change occurs. The sexual pattern appears to be essentially gonochoristic, but the potential for sex-change is not excluded. Received: 5 January 1999 / Accepted: 22 June 1999     

Biology of Serranicotyle labracis,ectoparasite of Dicentrarchus labrax (Teleostei): contribution to the study of its populations

Serranicotyle labracis (Van Beneden and Hesse, 1863) Maillard, Euzet and Silan, 1988 (synonym: Microcotyle labracis Van Beneden and Hesse, 1863) is a monogenean Polyopisthocotylea and a gill parasite of the bass Dicentrarchus labrax (Linné, 1758). Analysis of a western Mediterranean population of this parasite (Gulf of Lions, Languedoc, France) clarified several aspects of its biology. Its location in the gill biotope (Silan et al. 1987) was not fundamentally different from that found by Oliver (1977) and Winch (1983), but its distribution in host populations differed from that reported in the earlier studies; it displayed a negative binomial distribution in the study zone. At a finer level of observation, the behaviour of the hosts, as a function of degree of maturity and sex, appeared to determine more than any other factor the extent of infestation by the parasite. Previous suggestions in the literature to account for these differences in population dynamics are discussed.     

Structural, morphological and genetic variability in Halophila stipulacea (Hydrocharitaceae) populations in the western Mediterranean

Halophila stipulacea (Forssk.) Ascher. is a dioecious seagrass that colonized the Mediterranean basin probably following the opening of the Suez Canal (1869). Natural meadows have been reported since the end of the last century on the eastern side of the basin and only recently along the northern coast of Sicily. In the present study we examined the morphological and genetic variability of two natural meadows located along the Sicilian coast (Vulcano Island and Oliveri-Tindari coastal lakes). In order to determine morphological and genetic polymorphism, samples were collected at different depths (5, 15 and 25 m depth at the Vulcano site) and positions (edge vs mid-bed) within the meadows. Statistically significant differences in phenotypic features were found between the factors “depth” and “position” within the same meadow and between the two localities. Genetic diversity was assessed using randomly amplified polymorphic DNA and found to be high. Deep and shallow stands of the Vulcano Island meadow clustered in different positions in the UPGMA tree. The shallow Vulcano stand was found to be closer to the shallow Oliveri-Tindari meadow than to the deeper stand from Vulcano. Mantel's test did not allow rejection of the null hypothesis of independence of morphological and molecular distance matrices. We conclude that (i) H. stipulacea shows high morphological and genetic polymorphism, (ii) environmental and/or ecological barriers exist between different depths, and (iii) trends of morphological and genetic variability may be influenced by different environmental and/or ecological factors. Received: 15 November 1998 / Accepted: 21 May 1999     

The role of sexual harassment in cave and surface dwelling populations of the Atlantic molly, Poecilia mexicana (Poeciliidae, Teleostei)

Sexual coercion is a common behaviour for males trying to compensate for being inferior in male competition and/or female choice. We measured the cost of male sexual harassment for females as reduced feeding time in three populations of the Atlantic molly (Poecilia mexicana). Of these populations, one originated from a typical river habitat, another from a lightless cave chamber and the third one from a small, milky creek outside the cave. We gave hungry females an opportunity to feed in the presence of either another female or a male. We found the following: (1) male sexual harassment represents a cost in the river dwelling population and both small and large males significantly reduce female feeding time; and (2) sexual harassment was not detected in the other two populations (cave and cave entrance). In the cave molly, small males are at a disadvantage in female choice, but predation by an aquatic heteropteran selects against large male body size.     

Mitochondrial evolution and phylogeography in the hydrozoan<Emphasis Type="Italic"> Obelia geniculata</Emphasis> (Cnidaria)

The distribution and genetic structure of many marine invertebrates in the North Atlantic have been influenced by the Pleistocene glaciation, which caused local extinctions followed by recolonization in warmer periods. Mitochondrial DNA markers are typically used to reconstruct species histories. Here, two mitochondrial markers [16S rDNA and cytochrome c oxidase I (COI)] were used to study the evolution of the widely distributed hydrozoan Obelia geniculata (Linnaeus, 1758) from the North Atlantic and the Pacific and, more specifically, in the context of North Atlantic phylogeography. Samples were collected from six geographic localities between 1998 and 2002. Hydroids from the North Atlantic, North Pacific (Japan), and South Pacific (New Zealand) are reciprocally monophyletic and may represent cryptic species. Using portions of the 16S rDNA and COI genes and the date of the last trans-Arctic interchange (3.1–4.1 million years ago), the first calibrated rate of nucleotide substitutions in hydrozoans is presented. Whereas extremely low substitution rates have been reported in other cnidarians, mainly based on anthozoans, substitution rates in O. geniculata are comparable to other invertebrates. Despite a life history that ostensibly permits substantial dispersal, there is apparently considerable genetic differentiation in O. geniculata. Divergence estimates and the presence of unique haplotypes provide evidence for glacial refugia in Iceland and New Brunswick, Canada. A population in Massachusetts, USA, appears to represent a relatively recent colonization event.Communicated by J.P. Grassle, New Brunswick     

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     

Demographic parameters of two populations of red coral (Corallium rubrum L. 1758) in the North Western Mediterranean

The demographic and reproductive structure of populations represents the main data set needed for conservation and management plans. Mediterranean red coral Corallium rubrum has been exploited for 2,000 years, but only recently management plans have been solicited by the international community. We examined and compared the demographic features of two red coral shallow populations located in distinct geographic locations: Portofino (Italy) and Cap de Creus (Spain). Adults and juveniles density, growth rates, population size and age structure, fecundity and fertility were examined. Juveniles were the dominant class (33 %) in both populations. The analysis of the gamete content of 653 colonies revealed that the populations have balanced sex ratios and similar fertility and polyp fecundity. The average annual growth rate, determined on 119 colonies by annual growth rings count, was similar in both populations (0.24 mm year^?1), decreasing with colony age. Maximum life span of 99 % of the colonies was 60 and 40 years at Portofino and Cap de Creus, respectively. Minimum harvestable size (7 mm basal diameter) was reached in 30–35 years, and the percentage of colonies above it was 6.7 % at Portofino and 2.1 % at Cap de Creus, where juvenile and adult colony densities were significantly lower and the percentage of commercial-sized colonies reduced by 25 % in just a few years. Notwithstanding similar growth and fecundity, the two populations showed different densities and size/age structures suggesting local factors, together with different fishing pressures, have to be taken into account in the management plans for this species.     

Genetic protein variation in red mullet (Mullus barbatus) and striped red mullet (M. surmuletus) populations from the Mediterranean Sea

Starch-gel electrophoresis of allozymes was used to differentiate the two red mullet species (Mullus barbatus L. and M. surmuletus L.) in the Mediterranean Sea and, further, to investigate the genetic stock structure of M. barbatus in the eastern Mediterranean area. Twenty putative enzyme-coding loci were examined in eight M. barbatus samples caught in the Aegean and Ionian Seas (Greece) and in the Gulf of Lion (France), and two M. surmuletus samples caught in the Aegean and Gulf of Lion. A high degree of genetic polymorphism was found in both species. Species-specific electrophoretic patterns were found in PGI* and PGM*. Estimates of variance of allele frequencies among samples (F _ST) and ² analyses both revealed significant differences (P < 0.05) among the M. barbatus samples. Most of the genetic variation was among samples regardless of region. The mean value of Nei's genetic distance between the two species was 0.329. Genetic distance among M. barbatus samples was low (maximum Nei's D = 0.012), with the sample from Platania differing most from other M. barbatus samples. This is probably be due to founder effects existing at this area. These results suggest that allozyme analysis may provide important information on the genetic structure of the red mullet to ensure sustainable management of this species. Received: 7 May 1997 / Accepted: 13 October 1997     

Predator avoidance and school size in a cyprinodontid fish,the banded killifish (Fundulus diaphanus Lesueur)

Summary Solitary and schooling banded killifish (Fundulus diaphanus Lesueur) were attacked with a fish predator model in the littoral zone of a lake and their flight reaction distance to the model recorded. Reaction distance of solitary fish did not differ significantly from that of schooling fish, which was statistically constant over a wide range of observed school sizes. The results fo not support the proposed early predator warning function of social groups, which predicts an asymptotically increasing flight reaction distance with increasing group size. However, fish in larger and safer groups may detect an approaching predator sooner but delay their flight response, thereby permitting their individual members to spend more time assessing the nature of the threat before escaping. Such a strategy would reduce costly false alarms. Relative variation in recorded reaction distances was correlated negatively with school size, suggesting that schooling reduces variability in latency time to escape predators. This may be evidence for the above behavioural strategy. Further, the speed of transmission of the flight response within the school greatly exceeded the speed of approach of the predator model. This phenomenon enables schooling individuals to initiate avoidance behaviour before the approaching predator can be detected and before it reaches them.     

Allozyme and mitochondrial DNA variation in orange roughy,Hoplostethus atlanticus (Teleostei: Trachichthyidae): little differentiation between Australian and North Atlantic populations

Allozyme and mitochondrial DNA (mtDNA) genetic variation was compared in orange roughy (Hoplostethus atlanticus Collett) collected from waters off southern Australia and from waters about 22 000 km away in the North Atlantic west of Scotland. Samples were screened for 11 polymorphic allozyme loci and with 9 restriction enzymes. Significant heterogeneity between the two areas was detected for three allozyme loci (ADA ^*, CK ^* and GPI-1 ^*), and the overall G _ST (gene-diversity statistic) value of 1% was small but significant. Significant mtDNA haplotype heterogeneity was observed after ²- of haplotype frequencies but not after a G _ST analysis. Nucleotide sequence-diversity analysis showed very low net divergence (0.0023%) between the two samples. The Australian orange roughy had a lower allozyme heterozygosity and a lower mitochondrial DNA nucleon diversity than the North Atlantic sample. The very limited, although significant, allozyme and mitochondrial DNA heterogeneity between these areas suggests that there is some gene flow between these two populations. The species appears to be widespread, with its presence reported from the southern Pacific, southern Indian, and northern and southern Atlantic Oceans, and it is likely that gene flow between the antipodes is mediated by stepping-stone exchange between adjacent populations rather than by direct migration.     

Mitochondrial DNA analysis of the genetic relationships among populations of scad mackerel (Decapterus macarellus, D. macrosoma, and D. russelli) in South-East Asia

The genetic relationships among South-East Asian populations of the scad mackerels Decapterus macarellus, D. macrosoma and D. russelli (Pisces: Carangidae) were investigated. In 1995 and 1996, 216 fish were sampled in seven localities spanning the seas of Indonesia and were examined for restriction-site polymorphisms using ten restriction enzymes for the mitochondrial (mt) DNA control region, amplified by the polymerase chain-reaction. The inferred phylogeny of haplotypes led to the recognition of three distinct mitochondrial lineages or phylads consistent with the distinctions of current taxonomy. All 15 mtDNA haplotypes found in D. macarellus and all 9 haplotypes found in D. macrosoma were arranged as star-like clusters, suggesting recent evolutionary history. In contrast, the phylad formed by 6 haplotypes in D. russelli from the Sulawesi Sea exhibited diffuse topology, suggesting that ancestral lineages of this species have been retained to the present. Average nucleotide-divergence estimates between haplotypes of different phylads were between 0.042 and 0.135, suggesting ancient separation, in consistency with published allozyme data. High levels of haplotype diversity, but no geographical heterogeneity, was detected within D. macarellus from the Molucca Sea and the Banda Sea. Populations of D. macrosoma exhibited both significant differences between adjacent regions (Sunda Strait and Java Sea), and broadscale genetic homogeneity from the South China Sea to the Sulawesi Sea via the Java Sea and Makassar Strait. The geographic isolation of the D. macrosoma population sampled in the Sunda Strait suggests that this region constitute a sharp transition zone between the Indian Ocean and the Sunda Shelf. Near-monomorphism of haplotypes and low nucleotide diversity (d _X) were observed in the samples of D. macrosoma from the continental shelf (haplotype-diversity estimates, h, = 0.00 to 0.25 ± 0.08 and d _X = 0.000 to 0.002). This was in contrast to the comparatively high haplotype and nucleotide diversities observed in other pelagic fish species including D. macarellus (h = 0.82 ± 0.05, d _X = 0.012 to 0.015) and D. russelli (h = 0.63 ± 0.12, d _X = 0.016), and in the oceanic D. macrosoma population sampled in the Sunda Strait (h = 0.67 ± 0.31, d _X = 0.005). We hypothesise that this may be the consequence of recent and perhaps repeated bottleneck events that have affected the D. macrosoma population sampled on the continental shelf. Received: 29 September 1997 / Accepted: 3 September 1999     

Female discharges are more electrifying: spontaneous preference in the electric fish,Eigenmannia (Gymnotiformes,Teleostei)

Summary The tropical South American teleost Eigenmannia lineata showed a spontaneous preference for the female type, compared with the male type, of its sexually dimorphic, weak-electric organ discharge (EOD). Female and male EODs differ in waveform and harmonic content. An isolated fish was simultaneously stimulated with digitally synthesized natural male and female EODs of equal peak-to-peak amplitudes, at ±35 Hz frequency difference centered on its stable resting discharge frequency. The stimulus dipoles were arranged symmetrically to the right and left of the fish's hiding place. All stimulus conditions were permuted at random sequence. Among 11 fish tested, 8 showed a statistically significant preference for one stimulus, the female type, as measured by the amount of time a fish spent close to a stimulus dipole (P<0.05 in each fish, two-tailed). Thus female EODs rather than male EODs were more attractive to adult and juvenile fish of both sexes. It was also concluded that E. lineata is capable of discriminating female from male EODs by a complex sensory capacity requiring neither amplitude nor frequency cues. The EOD waveform changed very little within the ecological range of water conductivities (approximately 10–100 S·cm^-1); the P/N-ratio (a waveform character based on zerocrossing intervals) depended only weakly, but significantly, on conductivity (negative correlation in all four fish). Also, the effect of temperature on EOD waveform was very weak: Q ₁₀-values of the P/N-ratio were below but close to 1 in all fish (27±5°C). Thus, it can be concluded that the EOD waveform is remarkably stable within widely changing conditions-even beyond the variation found in the field-and is therefore potentially useful as a social cue.     

Microphytoplankton in the Strait of Otranto (eastern Mediterranean)

The Strait of Otranto is the connection between the Adriatic and Ionian Seas. Low nutrient concentrations, high transparency, and low phytoplankton cell density and biomass reflect the oligotrophic character of the area. Enrichment of the euphotic layer with nutrients is mainly due to discharge of Albanian and Greek rivers, as well as mixing and upwelling in winter/early spring. Following phytoplankton bloom in April, a progressive decrease of phytoplankton cell density is due to the consumption of nutrients throughout the proceeding summer and autumn. Nitrogen was a strong limiting factor for phytoplankton growth in summer. Deep biomass maxima were detected in the 50 to 100 m layer and corresponded mostly to cells smaller than 20 m. The eastern part of the strait is mostly influenced by the northerly inflowing current from the Ionian Sea, and the western part by the southerly outflowing current from the Adriatic Sea. This typical circulation could be disturbed by inertial oscillations in the current field, generated by the strong oscillating winds and cyclonic eddies. The type of circulation determined the distribution of thermohaline characteristics, abundance, biomass, as well as taxonomic composition of phytoplankton, across the strait. Ecological characteristics of the water masses on two sides of the strait were significantly different during the formation of a longitudinal thermohaline front in May 1990.     

Histology and ultrastructure of the skin of Lepadichthys lineatus (Gobiesocidae: Teleostei)

The skin of the Red Sea clingfish Lepadichthys lineatus Briggs, 1966, which lives on shallow-water crinoids, consists of a continuous layer of giant mucus-producing cells. Upon stimulation, these cells are able, in seconds, to envelope the fish entirely with a thick layer of their secretion. The cells are 75 to 290 high, and are enveloped by septa of compressed epithelium. They rest on a strong collagenous layer, and extend to the skin surface, where they open. The nuclei of the giant cells are large, and irregularly shaped; their cytoplasm has a very extensive endoplasmic reticulum and undergoes deep structural changes during maturation. Their ability to produce large amounts of mucus seems to be an adaptive device to protect the fish from contact with the rough surface of the crinoid host.