TWO SOLIERIACEAN ALGAE NEW TO THE MEDITERRANEAN: AGARDHIELLA SUBULATA AND SOLIERIA FILIFORMIS (RHODOPHYTA,GIGARTINALES)1

Two Atlantic species found in the Mediterranean Sea were ascribed to the family Solieriaceae on the basis of morphological features. In the extensive drifting beds of the Mar Piccolo basin (Taranto, Ionian Sea), Agardhiella subulata (C. Agardh) Kraft et Wynne and Solieria filiformis (Kützing) Gabrielson were the dominant species in summer. There were no attached populations, and perennation was ensured only by vegetative propagation. Characteristics of the outer cortex and chromatophores were useful for distinguishing the taxa at the species level and are recommended for recognition of sterile specimens.     

Structure and function of a mating-type gene from the homothallic species Neurospora africana

The homothallic Neurospora species, N. africana, contains sequences that hybridize to the A but not to a mating-type sequences of the heterothallic species N. crassa. In this study, the N. africana mating-type gene, mt A-1, was cloned, sequenced and its function analyzed in N. crassa. Although N. africana does not mate in a heterothallic manner, its mt A-1 gene functions as a mating activator in N. crassa. In addition, the N. africana mt A-1 gene confers mating type-associated vegetative incompatibility in N. crassa. DNA sequence analysis shows that the N. africana mt A-1 open reading frame (ORF) is 93% identical to that of N. crassa mt A-1. The mt A-1 ORF of N. africana contains no stop codons and was detected as a cDNA which is processed in a similar manner to mt A-1 of N. crassa. By DNA blot and orthogonal field agarose gel electrophoretic analysis, it is shown that the composition and location of the mating-type locus and the organization of the mating-type chromosome of N. africana are similar to that of N. crassa.     

Review of Atlantoscia Ferrara & Taiti, 1981 (Crustacea: Isopoda: Oniscidea: Philosciidae) with new records and new species

The neotropical genus Atlantoscia Ferrara & Taiti, 1981 includes three species, A. floridana Van Name, 1940, occurring in coastal regions of Florida (USA), Brazil, Argentina, and Ascension and Saint Helena islands, A. rubromarginata Araujo & Leistikow, 1999 and A. petronioi Campos-Filho, Contreira and Lopes-Leitzke, 2012 from Brazil. Two new species from Brazil are here described, one from the state of Bahia and one from the state of São Paulo. All the new species have respiratory areas on pleopods 1–5 exopods. The specimens were collected in areas of Atlantic Forest and in cocoa (Theobroma cacao) plantations. New records of A. floridana and A. rubromarginata and distribution maps are presented.     

Convergent evolution of morphological and ecological traits in the open-habitat chat complex (Aves, Muscicapidae: Saxicolinae)

Snails in the closely related trochid genera Phorcus Risso, 1826 and Osilinus Philippi, 1847 are ecologically important algal grazers in the intertidal zone of the northeastern Atlantic Ocean and Mediterranean Sea. Here we present the first complete molecular phylogeny for these genera, based on the nuclear 28S rRNA gene and the mitochondrial 16S rRNA and COI genes, and show that the current classification is erroneous. We recognize nine species in a single genus, Phorcus: estimated by BEAST analysis, this arose 30 (±10) Ma; it consists of two subgenera, Phorcus and Osilinus, which we estimate diverged 14 (±4.5) Ma. Osilinus kotschyi, from the Arabian and Red Seas, is not closely related and is tentatively referred to Priotrochus Fischer, 1879. Our phylogeny allows us to address biogeographical questions concerning the origins of the Mediterranean and Macaronesian species of this group. The former appear to have evolved from Atlantic ancestors that invaded the Mediterranean on several occasions after the Zanclean Flood, which ended the Messinian Salinity Crisis 5.3 Ma; whereas the latter arose from several colonizations of mainland Atlantic ancestors within the last 3 (±1.5) Ma.     

Ecological characterization of entomopathogenic nematodes isolated in stone fruit orchard soils of Mediterranean areas

Entomopathogenic nematodes in the families Steinernematidae and Heterorhabditidae were isolated from stone-fruit orchards in two Mediterranean regions of Spain. A total of 630 soil samples (210 sites) from Catalonia and 90 soil samples (30 sites) from Murcia were evaluated resulting in 5.2% and 20% of the soils testing positive for nematodes, respectively. Ten steinernematid isolates and three heterorhabditid isolates were recovered using the Galleria mellonella baiting method. Based on morphometric data, molecular data, and cross-breeding experiments the nematode species were identified as Steinernemafeltiae and Heterorhabditis bacteriophora. Environmental tolerance to heat, desiccation and hypoxia, the effect of temperature on infectivity and reproduction and nematode migration in sand columns were compared among isolates and one Steinernema carpocapsae strain. Results showed differences among species and a great variability within species. Beneficial traits for each strain were added up to identify a superior candidate to control Mediterranean flat-headed rootborer, Capnodis tenebrionis. When all analyzed factors were considered, three S. feltiae isolates (Bpa, Sor and M116) obtained the best scores, and when hypoxia was removed, two of the strains (Bpa and Sor) continued ranking superior to other strains.     

Identification and Molecular Characterization of the Homogentisate Pathway Responsible for Pyomelanin Production,the Major Melanin Constituents in Aeromonas media WS

The pigmentation of many Aeromonas species has been thought to be due to the production of a L-DOPA (L-3,4-dihydroxyphenylalanine) based melanin. However, in this study we found that although L-DOPA synthesis occurs in the high-melanin-yielding Aeromonas media strain WS, it plays a minor, if any, role in pigmentation. Instead, the pigmentation of A. media strain WS is due to the production of pyomelanin through HGA (homogentisate). Gene products of phhA (encodes phenylalanine hydroxylase), tyrB and aspC (both encode aromatic amino acid aminotransferase), and hppD (encodes 4-hydroxyphenylpyruvate dioxygenase) constitute a linear pathway of converting phenylalanine to HGA and disruption of any one of these genes impairs or blocks pigmentation of A. media strain WS. This HGA biosynthesis pathway is widely distributed in Aeromonas, but HGA is only detectable in the cultures of pigmented Aeromonas species. Heterologous expression of HppD from both pigmented and non-pigmented Aeromonas species in E. coli leads to the production of pyomelanin and thus pigmentation, suggesting that most Aeromonas species have the critical enzymes to produce pyomelanin through HGA. Taken together, we have identified a widely conserved biosynthesis pathway of HGA based pyomelanin in Aeromonas that may be responsible for pigmentation of many Aeromonas species.     

Molecular evidence of male-biased dispersal in loggerhead turtle juveniles

Serum testosterone levels and mtDNA haplotypes were obtained from 65 juvenile loggerhead turtles (Caretta caretta, L.) incidentally caught in the central Mediterranean. The group of specimens carrying a haplotype specific for the northwest Atlantic had higher testosterone levels, and so included more males, than the other one. Since primary sex ratios of northwest Atlantic colonies are strongly skewed towards females, results indicate a male bias among Atlantic turtles entering the Mediterranean. This demonstrates for the first time a sex-biased dispersal of specimens in the pelagic phase, an important factor to be considered in conservation programs.     

The salinity preference of members of the genus Macroplea (Coleoptera, Chrysomelidae, Donaciinae), fully aquatic leaf beetles that occur in brackish water

Reed beetles (Donaciinae) of the genus Macroplea Samouelle, 1819 live permanently submerged. Literature indicates that Macroplea mutica occurs in brackish water, whereas Macroplea appendiculata is restricted to freshwater. The salinity preference of these two species was tested in a linear and a circular device that offered a continuous salinity gradient. The distribution of animals in the devices was monitored over at least 3 h in each of the 21 experiments. Both species preferred freshwater (salinity 0) over brackish water (salinity 10). In particular, this holds true for specimens collected in brackish water. Likewise, immediate reactions could be observed when during such experiments the direction of the gradient was reversed. While M. mutica can be regarded as a truly marine insect, this marine environment does not strictly reflect its fundamental niche with respect to salinity preference. This is in line with accumulating evidence that M. mutica can be found in freshwater habitats (and M. appendiculata in brackish water). This indicates that the species’ distribution might be influenced by other factors like host plant preference or dispersal mechanisms. It is discussed if—in spite of similar fundamental niches—differences in salinity tolerance (and hence performance in brackish water) may have contributed to speciation in the genus Macroplea.     

New fossil records of Tapirus (Mammalia, Perissodactyla) from Brazil, with a critical analysis of intra-generic diversity assessments based on lower molar size variability

A large set of South American fossils belonging to the genus Tapirus has been described on the basis of differences in size and proportions of lower molariform teeth. Nevertheless, the reliability of dental proportions for the diagnosis of fossil tapir species is controversial. In this paper, we describe new fossil material of Tapirus from the Quaternary of Serra da Bodoquena, Southwestern Brazil, comparing it to other fossil and extant specimens of the genus by means of multivariate morphometric analyses of lower molariform teeth linear dimensions. The results of Principal Component Analyses indicate that some of the extant and extinct material attributed to Tapirus fall within the range of variation in size and proportions of lower molariform teeth exhibited by recent species of the genus. Therefore, part of the fossil material attributed to new species or to Tapirus sp. may be referable to the extant species Tapirus terrestris. We conclude that the sole use of lower molariform teeth size and proportions to erect new species of Tapirus may not be reliable, and therefore we advocate caution when describing fossil tapirs exclusively based on these criteria.     

Invasion by the marine gastropod Ocinebrellus inornatus in France: I. Scenario for the source of introduction

The rate of introduction of exotic marine species has dramatically increased during the 19th and 20th centuries. Exemplifying this trend, the marine gastropod Ocinebrellus inornatus was first detected outside its native range in 1924 on the American Pacific coast, then in 1995 on the French Atlantic coast. To determine the origin of the French populations of this invasive species, we compared a French population with populations collected in Asia—the native range—and with a population collected in the United States. Analyses of mitochondrial DNA and allozyme polymorphism revealed that the French and American populations were closely related and substantially differentiated from the Asian populations. According to our results, the most likely scenario is that the source population of the French Atlantic coast populations was located in the United States. Indeed, taken altogether, the genetic structure of Asian populations, the time lag separating the introduction on the American Pacific coast from the introduction on the French Atlantic coast and the high level of genetic diversity in the two introduced areas (indicating an absence of major founder events) are hardly compatible with a scenario in which French population resulting only from primary introduction events from the native area. Finally, although similar, the French and American populations were not identical. Thus, even if the main source population of the French populations was located in the United States, the genetic structure of French populations may have been modified by cryptic and recurrent introduction events directly from Asia.     

Characterization of the bacterial communities associated with the bald sea urchin disease of the echinoid Paracentrotus lividus

The microbial communities involved in the bald sea urchin disease of the echinoid Paracentrotus lividus are investigated using culture-independent techniques. Lesions of diseased specimens from two locations in France, La Ciotat (Mediterranean Sea) and Morgat (Atlantic Ocean), are examined by Scanning Electron Microscopy (SEM) and the diversity of their microbiota is analysed by Denaturing Gradient Gel Electrophoresis (DGGE) and 16S rRNA gene clones libraries construction. Microscopic observations demonstrated that only the central area of the lesions is invaded by bacteria but not the peripheral zone and the surrounding healthy tissues. Molecular analysis identified at least 24 bacterial genomospecies in bald sea urchin lesions: 5 are Alphaproteobacteria, 10 are Gammaproteobacteria, 8 are CFB bacteria and 1 is a Fusobacteria. Out of them, 4 are observed in both locations while 10 occur only in the Atlantic Ocean and 10 only in the Mediterranean Sea. Gammaproteobacteria are the most represented in clones libraries from both locations, with respectively 65% and 43% of the total clones. CFB and Alphaproteobacteria accounted for the majority of the remaining clones and were detected by DGGE in virtually all samples from both stations. Our results demonstrate that bacterial communities observed on diseased individuals of the same echinoid species but originating from distinct locations are not similar and thus support the hypothesis that bacteria involved in the worldwide echinoid disease commonly called the bald sea urchin disease are opportunistic and not specific.     

Morphological Differentiation Between Geographically Separated Populations of <Emphasis Type="Italic">Neomysis integer</Emphasis> and <Emphasis Type="Italic">Mesopodopsis slabberi</Emphasis> (Crustacea,Mysida)

Morphological variation was examined in Neomysis integer and Mesopodopsis slabberi, two abundant, low dispersal mysid species (Crustacea, Mysida) along the European coasts. Both species dominate the hyperbenthic communities in the northeast Atlantic, and M. slabberi is also widely distributed in the Mediterranean and Black Sea. Three populations of these species were sampled throughout their distribution range; samples of N. integer were collected in the northeast Atlantic Eems-Dollard, Gironde and Guadalquivir estuaries; in the case of M. slabberi, mysids were sampled in two northeast Atlantic estuaries (Eems-Dollard and Guadalquivir) and one Mediterranean site (Ebro Delta). A total of 12 morphometric and 2 meristic characters were measured from 30–64 mysids per sample. Multivariate analysis showed clear morphometric differences between populations of both species. The morphological differentiation within M. slabberi was highly concordant with the available genetic data from mitochondrial loci, pointing to a large divergence between the Atlantic and Mediterranean populations. However, due to some overlap of individuals between the different populations, the present morphometric analysis does not suffice to assign the different populations to a separate (sub)species status. In the case of N. integer, the morphometric patterns showed a divergence of the Gironde population. Differentiation of populations within this mysid, as in M. slabberi, were mainly related to eye and telson morphology. Potential interactions of the mysid morphology and environmental conditions are discussed.     

Octopus vulgaris (Cuvier, 1797) in the Mediterranean Sea: Genetic Diversity and Population Structure

The common octopus, Octopus vulgaris Cuvier 1797, is a largely exploited cephalopod species in the Mediterranean Sea and the Atlantic Ocean, as well as along the coasts of Africa, Brazil and Japan, where its taxonomic identity is still debated. The assessment of its genetic structure is a pressing need to correctly manage the resource and to avoid overfishing and collapsing of local stocks. Here we analysed genetic variation and population structure of O. vulgaris using thirteen microsatellite loci in seven sampling localities from the Mediterranean Sea and one from the Atlantic Ocean. We also used a DNA barcoding approach by COI gene fragment to understand the phylogenetic relationships among the specimens here investigated and the ones whose sequences are available in literature. Our results reveal high levels of allelic richness and moderate heterozygosity in all samples investigated, and a pronounced differentiation of the Atlantic and Sicilian specimens. This latter aspect seems to support the isolation of the biota within the Strait of Messina. A certain degree of differentiation was detected among the other geographic samples within the Mediterranean Sea, which is more compatible with an island model than isolation by distance. The occurrence of null alleles affected more genetic diversity indices than population structure estimations. This study provides new insights about the genetic diversity and structure of O. vulgaris in the area of interest, which can be used as guidelines for a fisheries management perspective.     

The taxonomic status of some Atlanto‐Mediterranean species in the subgenus Holothuria (Echinodermata: Holothuroidea: Holothuriidae) based on molecular evidence

Molecular and morphological data were used to evaluate the taxonomic status of the species Holothuria tubulosa Gmelin, 1790 , Holothuria stellati Delle Chiaje, 1823 , Holothuria mammata Grube, 1840 , and Holothuria dakarensis Panning, 1939, belonging to the nominate subgenus Holothuria (Holothuria) (family Holothuriidae) from the Mediterranean Sea and Atlantic Ocean. A 16S rRNA marker distinguished three well‐supported clades with clear genetic differentiation amongst them. The morphometric characters, although they reflected the clades, showed great variability, and some specimens from different clades overlapped. The morphological data and the literature suggest that the clades correspond to H. dakarensis (from Cape Verde Islands), H. mammata (from the Atlanto‐Mediterranean area) and H. tubulosa (from the Mediterranean Sea). Holothuria stellati is considered here to be a junior subjective synonym of H. tubulosa. Great morphological intraspecific variation within H. tubulosa and H. mammata explains the confusion in the literature. Holothuria tubulosa includes specimens with distinctive ossicles, but others are similar to H. mammata. In these cases, the presence or absence of Cuvierian tubules proved a reliable indicator to the identity of these species; unfortunately this character is difficult to assess in preserved material. According to the results of discriminant analysis we propose a set of ossicle morphometric variables that permit the optimum assignation of individuals to the clades. Our results present a new perspective on the taxonomic status of species in Holothuria (Holothuria), and show how a molecular approach, combined with a morphological approach, can solve taxonomic problems.     

Natural or Naturalized? Phylogeography Suggests That the Abundant Sea Urchin Arbacia lixula Is a Recent Colonizer of the Mediterranean

We present the global phylogeography of the black sea urchin Arbacia lixula, an amphi-Atlantic echinoid with potential to strongly impact shallow rocky ecosystems. Sequences of the mitochondrial cytochrome c oxidase gene of 604 specimens from 24 localities were obtained, covering most of the distribution area of the species, including the Mediterranean and both shores of the Atlantic. Genetic diversity measures, phylogeographic patterns, demographic parameters and population differentiation were analysed. We found high haplotype diversity but relatively low nucleotide diversity, with 176 haplotypes grouped within three haplogroups: one is shared between Eastern Atlantic (including Mediterranean) and Brazilian populations, the second is found in Eastern Atlantic and the Mediterranean and the third is exclusively from Brazil. Significant genetic differentiation was found between Brazilian, Eastern Atlantic and Mediterranean regions, but no differentiation was found among Mediterranean sub-basins or among Eastern Atlantic sub-regions. The star-shaped topology of the haplotype network and the unimodal mismatch distributions of Mediterranean and Eastern Atlantic samples suggest that these populations have suffered very recent demographic expansions. These expansions could be dated 94–205 kya in the Mediterranean, and 31–67 kya in the Eastern Atlantic. In contrast, Brazilian populations did not show any signature of population expansion. Our results indicate that all populations of A. lixula constitute a single species. The Brazilian populations probably diverged from an Eastern Atlantic stock. The present-day genetic structure of the species in Eastern Atlantic and the Mediterranean is shaped by very recent demographic processes. Our results support the view (backed by the lack of fossil record) that A. lixula is a recent thermophilous colonizer which spread throughout the Mediterranean during a warm period of the Pleistocene, probably during the last interglacial. Implications for the possible future impact of A. lixula on shallow Mediterranean ecosystems in the context of global warming trends must be considered.     

Confirmation of neutral buoyancy in <Emphasis Type="Italic">Aethotaxis mitopteryx</Emphasis> DeWitt (Notothenioidei: Nototheniidae)

Aethotaxis mitopteryx is a nototheniid species with a circum-Antarctic distribution in the Southern Ocean. We present new locality records, buoyancy measurements, and morphometric data on five A. mitopteryx specimens collected from the Bransfield Strait. Our analyses demonstrate that A. mitopteryx is neutrally buoyant. This result lends additional support to the hypothesis that neutral buoyancy has a single evolutionary origin in Notothenioidei. Food items in these specimens consisted entirely of krill (Euphausia sp.), and all specimens were sexually immature. We present morphometric data from these five specimens and compare to the holotype specimen, which is the only A. mitopteryx specimen collected in the Ross Sea.     

Fickle or Faithful: The Roles of Host and Environmental Context in Determining Symbiont Composition in Two Bathymodioline Mussels

The Mediterranean Sea and adjoining East Atlantic Ocean host a diverse array of small-sized mussels that predominantly live on sunken, decomposing organic remains. At least two of these, Idas modiolaeformis and Idas simpsoni, are known to engage in gill-associated symbioses; however, the composition, diversity and variability of these symbioses with changing habitat and location is poorly defined. The current study presents bacterial symbiont assemblage data, derived from 454 pyrosequencing carried out on replicate specimens of these two host species, collected across seven sample sites found in three oceanographic regions in the Mediterranean and East Atlantic. The presence of several bacterial OTUs in both the Mediterranean Sea and eastern Atlantic suggests that similar symbiont candidates occur on both sides of the Strait of Gibraltar. The results reveal markedly different symbiotic modes in the two species. Idas modiolaeformis displays high symbiont diversity and flexibility, with strong variation in symbiont composition from the East Mediterranean to the East Atlantic. Idas simpsoni displays low symbiont diversity but high symbiont fidelity, with a single dominant OTU occurring in all specimens analysed. These differences are argued to be a function of the host species, where subtle differences in host evolution, life-history and behaviour could partially explain the observed patterns. The variability in symbiont compositions, particularly in Idas modiolaeformis, is thought to be a function of the nature, context and location of the habitat from which symbiont candidates are sourced.     

Structure and function of a mating-type gene from the homothallic species Neurospora africana

The homothallic Neurospora species, N. africana, contains sequences that hybridize to the A but not to a mating-type sequences of the heterothallic species N. crassa. In this study, the N. africana mating-type gene, mt A-1, was cloned, sequenced and its function analyzed in N. crassa. Although N. africana does not mate in a heterothallic manner, its mt A-1 gene functions as a mating activator in N. crassa. In addition, the N. africana mt A-1 gene confers mating type-associated vegetative incompatibility in N. crassa. DNA sequence analysis shows that the N. africana mt A-1 open reading frame (ORF) is 93% identical to that of N. crassa mt A-1. The mt A-1 ORF of N. africana contains no stop codons and was detected as a cDNA which is processed in a similar manner to mt A-1 of N. crassa. By DNA blot and orthogonal field agarose gel electrophoretic analysis, it is shown that the composition and location of the mating-type locus and the organization of the mating-type chromosome of N. africana are similar to that of N. crassa.     

Origin of the invasive Arundo donax (Poaceae): a trans-Asian expedition in herbaria

Background and Aims

The hypothesis of an ancient introduction, i.e. archaeophyte origin, is one of the most challenging questions in phylogeography. Arundo donax (Poaceae) is currently considered to be one of the worst invasive species globally, but it has also been widely utilzed by man across Eurasia for millennia. Despite a lack of phylogenetic data, recent literature has often speculated on its introduction to the Mediterranean region.

Methods

This study tests the hypothesis of its ancient introduction from Asia to the Mediterranean by using plastid DNA sequencing and morphometric analysis on 127 herbarium specimens collected across sub-tropical Eurasia. In addition, a bioclimatic species distribution model calibrated on 1221 Mediterranean localities was used to identify similar ecological niches in Asia.

Key Results

Despite analysis of several plastid DNA hypervariable sites and the identification of 13 haplotypes, A. donax was represented by a single haplotype from the Mediterranean to the Middle East. This haplotype is shared with invasive samples worldwide, and its nearest phylogenetic relatives are located in the Middle East. Morphometric data characterized this invasive clone by a robust morphotype distinguishable from all other Asian samples. The ecological niche modelling designated the southern Caspian Sea, southern Iran and the Indus Valley as the most suitable regions of origin in Asia for the invasive clone of A. donax.

Conclusions

Using an integrative approach, an ancient dispersion of this robust, polyploid and non-fruiting clone is hypothesized from the Middle East to the west, leading to its invasion throughout the Mediterranean Basin.