Bacterial symbionts as an additional cytological marker for identification of sponges without a skeleton

Symbiotic bacteria from six Oscarella species (adults and embryos) collected in the Mediterranean Sea (O. lobularis, O. tuberculata, O. imperialis, O. microlobata, O. viridis) and the Sea of Japan (O. malakhovi) were investigated by scanning electron microscopy and transmission electron microscopy. In most cases, symbionts are rather numerous. Each sponge species has a definite set of bacterial morphological types. All bacteria are extracellular. Symbionts occupy the mesohyl of adult sponges or intercellular space in embryos and are often in contact with mesohylar filaments or cells. Bacteria of some morphotypes have characteristic blebs. Most symbionts are gram-negative, and two types of bacteria have traits of Archaea and one type of bacteria is similar to Planctomycetes. Data on morphology of bacterial symbionts can be a good additional character for identification of Oscarella species, which have no skeleton.     

Transmission,plasticity and the molecular identification of cyanobacterial symbionts in the Red Sea sponge <Emphasis Type="Italic">Diacarnus erythraenus</Emphasis>

Cyanobacterial symbionts in the sponge Diacarnus erythraenus from the Red Sea were identified in both adult sponges and their larvae by 16S rDNA sequencing. A single cyanobacterial type was found in all samples. This cyanobacterial type is closely related to other sponge cyanobacterial symbionts. The cyanobacterial rDNA, together with the morphological analysis by electron and fluorescence microscopy, provided evidence for vertical transmission of the symbionts in this sponge. In addition, we show phenotypic plasticity of the symbionts inside the sponge, probably as a result of variability in light availability inside the sponge tissue. Finally, the reproduction of Diacarnus erythraenus is also described.Matan Oren, Laura Steindler have contributed equally to the work.     

Microbial associations in sponges. II. Numerical analysis of sponge and water bacterial populations

Three taxonomically distant sponges Pericharax heteroraphis, Jaspis stellifera and Neofibularia irata contain phenotypically similar bacterial symbionts which differ from bacteria in the ambient water. These symbionts are predominant in the sponges and were detected after computer analysis of 526 heterotrophic bacterial strains tested for 76 characters. These facultative anaerobic symbionts metabolize a wide range of compounds and may be important in removing waste products while the sponges are not circulating water. The bacteria produce sticky-mucoid colonies and thus would contribute to sponge structural rigidity. The fourth sponge Ircinia wistarii contains a mixed aerobic population similar to that in the ambient water. The majority of the bacteria are located around the inhalant canals, facilitating the uptake of dissolved organic matter and oxygen from the incoming water.     

Susceptibility to oxidative stress of the Mediterranean demosponge Petrosia ficiformis : role of endosymbionts and solar irradiance

 The effects of elevated pO₂ and irradiance as inducers of prooxidant conditions have been investigated in the Mediterranean demosponge Petrosia ficiformis (Poiret, 1789). This species lives symbiotically with the autotrophic cyanobacterium Aphanocapsa feldmanni, the abundance of which is controlled by the intensity of light irradiance. In the presence of symbionts, tissues of P. ficiformis were characterized by a general enhancement of antioxidant defenses as compared to aposymbiotic specimens. The main differences included higher activities of several antioxidant enzymes and a greater capability to neutralize various forms of oxyradicals, as indicated by the total oxyradical scavenging capacity (TOSC) assay. Elevated pO₂, more than light, appeared to be the primary factor inducing prooxidant pressure in the Mediterranean sponge; in fact, irrespective of the solar irradiance experienced by the sponge, symbiotic specimens showed comparable activities of antioxidant enzymes and a similar scavenging capacity towards various reactive oxygen species. However, the potential toxicity of photodynamic production of reactive oxygen species was demonstrated in organisms from more irradiated sites, as the levels of antioxidant defenses were lowered in the outer layer of the sponge. The role of enhanced antioxidant defenses in protecting symbiotic specimens, also from oxyradical-mediated toxicity of light exposure, was supported by translocation experiments; aposymbiotic sponges did not survive when moved to conditions of elevated solar irradiance, while no effects were observed in symbiotic specimens if translocated and/or deprived of symbionts. Received: 23 November 1999 / Accepted: 13 June 2000     

Cellular localisation of secondary metabolites isolated from the Caribbean sponge <Emphasis Type="Italic">Plakortis simplex</Emphasis>

The Caribbean sponge, Plakortis simplex, is known to contain a large array of secondary metabolites, including the antimalarial polyketide plakortin, several unusual glycolipids, and some hopanoids, which closely resemble typical bacterial metabolites. The hypothesis that they could be products of bacterial metabolism was tested by localizing specific metabolites in cells using physical separation of sponge cells, bacterial symbionts and supernatant by differential centrifugation. The obtained fractions were analysed separately for the typical P. simplex metabolites by NMR and mass spectrometry, and most of them were shown to be present in the bacterial cells but not in the sponge cells. In addition, PCR screening showed that the biosynthetic pathway for glycosphingolipids was present in the bacterial cells. Isolation of a Sphingomonas strain PS193 from P. simplex and subsequent glycosphingolipid analysis resulted in the detection of a known glycosphingolipid, GSL-1, that did, however, not match the glycosphingolipid profile of P. simplex. Therefore, it is unlikely that Sphingomonas strain PS193 is an abundant member of the microbial community associated with P. simplex. Other glycosphingolipid producing bacteria in P. simplex remain to be identified. In conclusion, this study provides experimental evidence that the glycolipids and hopanoids and possibly also the polyketide plakortin are produced by microbial symbionts rather than the sponge from which the metabolites were originally isolated.     

Host specificity of the symbiotic cyanobacterium<Emphasis Type="Italic"> Oscillatoria spongeliae</Emphasis> in marine sponges,<Emphasis Type="Italic"> Dysidea</Emphasis> spp.

Marine sponges can host a variety of cyanobacterial and bacterial symbionts, but it is often unclear whether these symbionts are generalists that occur in many host species or specialists that occur only in certain species or populations of sponges. The filamentous cyanobacterium Oscillatoria spongeliae is found in the sponges Dysidea n. sp. aff. herbacea 1A and 1B, and similar cyanobacteria are found in D. n. sp. aff. granulosa. We amplified and sequenced sponge nuclear ribosomal DNA (rDNA) and cyanobacterial 16S rDNA from specimens of these three sponges. We then used these sequences to construct phylogenies for host sponges and their symbiotic cyanobacteria. Each of these three sponge species hosts a distinct cyanobacterial clade, suggesting a high degree of host specificity and potential coevolution between symbiotic cyanobacteria and their host sponges.     

Metabolic integration between symbiotic cyanobacteria and sponges: a possible mechanism

Metabolic relationships between symbiotic cyanobacteria and host sponge have been investigated in the marine species Chondrilla nucula and Petrosia ficiformis (collected in the Ligurian Sea in 1992). DNA, RNA, total protein, cytosolic protein, total sugar, cytosolic sugar, total lipid, nonprotein sulfhydryl groups, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase were assayed in cortex-free sponge tissue, where cyanobacteria are all but absent. For both species, biochemical parameters were determined in specimens living in illuminated habitats and in dark caves, where sponges are virtually aposymbiotic for cyanobacteria. As C. nucula is unable to colonize dark sites, specimens of this species were artificially transferred to a cave and maintained in dark conditions for 6 mo. Results showed that in the absence of light (i.e., in the absence of cyanobacteria) C. nucula undergo metabolic collapse and thiol depletion. In contrast, P. ficiformis activates heterotrophic metabolism and mechanisms which balance the loss of cell reducing power. This suggests that cyanobacteria effectively participate in controlling the redox potential of the host cells by the transfer of reducing equivalents. Only P. ficiformis is capable of counteracting, by means of heterotrophic metabolism, the loss of the contribution from symbionts which is caused by dark conditions. This explains the differences in the ecological requirements of the two species. Because cyanobacterial symbionts release fixed carbon in the form of glycerol and other small organic phosphate (Wilkinson 1979), a model based on the glycerol 3-phosphate shuttle (typically occurring in chloroplasts and mitochondria) is suggested. The mechanism proposed appears to be an ancient biochemical adaptation which arose among ancestral symbiotic systems, and further developed in the relationships between endosymbiotic organelles and cytoplasm.     

Symbiotic zooxanthellae enhance boring and growth rates of the tropical sponge Anthosigmella varians forma varians

Several species of boring sponges harbor symbiotic zooxanthellae, and it is believed that the symbiont enhances boring activity of host sponges. This hypothesis was tested using manipulative field experiments to assess the effect of intracellular zooxanthella populations on boring rates of the tropical sponge Anthosigmella varians forma varians. Portions of sponge were attached to 60 calcium carbonate blocks of known weight. Three sets of 10 blocks were grown at high light levels and three sets of 10 blocks were grown at low light levels for 105 d in the Florida Keys, Florida, USA. Boring rates, growth rates (lateral growth and within-substratum tissue penetration), and zooxanthella populations were measured at the end of the experiment. Absolute rates of boring and growth of A. varians forma varians were significantly greater when zooxanthella densities were higher. Boring rate and tissue penetration related to final surface area of sponge attachment was also enhanced when zooxanthella densities were higher, suggesting that the symbiont plays a physiological role in the decalcification process. This is in contrast to the role that zooxanthellae play in coral hosts. Based on the results of this study, it appears that the presence of zooxanthellar symbionts has important ecological and life-history consequences for host sponges. Ability to laterally overgrow competitors will be correlated with the size and activity of zooxanthella populations. In addition, the fitness of host sponges will be enhanced by algal symbionts, since greater penetration within substrata will result in an increase in production of tissue that can be converted into storage, feeding and reproductive functions.     

Innate immune defense of the sponge<Emphasis Type="Italic"> Suberites domuncula</Emphasis> against gram-positive bacteria: induction of lysozyme and AdaPTin

Sponges are filter feeders that are exposed to large amounts of bacteria present in their surrounding aqueous milieu. The characteristic cell wall component of gram-positive bacteria, peptidoglycan (PPG), was used as a model molecule to study the responsiveness of cells from the marine demosponge Suberites domuncula toward gram-positive bacteria. The sponge lysozyme, which hydrolyzes PPG, was isolated from the living sponge; in addition its gene was cloned (SDLYS) and expressed in Escherichia coli. Antibodies were raised against the recombinant protein to demonstrate that in the Western blot both molecules give the same signal. In situ hybridization with SDLYS as a probe showed that cells in the mesohyl, the gray cells, strongly react with SDLYS. Subsequent immunofluorescence studies with antibodies raised against lysozyme revealed that only bacteria react with anti-lysozyme and only those that are scattered within the mesohyl of the tissue. An adaptor gene (AdaPTin-1) was isolated from the same sponge species that encodes a putative protein involved in endosome formation. Based on its differential expression we conclude that sponge cells react to PPG with a rapid activation of endocytosis, followed by the release of lysozyme.Communicated by O. Kinne, Oldendorf/LuheThe cDNA sequences from Suberites domuncula have been deposited (EMBL/GenBank): cDNA for the lysozyme (SDLYS) under the accession number AJ699166 and the AdaPTin-1 cDNA (SDAP1) under AJ699167.     

Biochemical genetic divergence and systematics in sponges of the genera Corticium and Oscarella (Demospongiae: Homoscleromorpha) in the Mediterranean Sea

The sponge sub-class Homoscleromorpha is generally considered to include just two families, the Oscarellidae (without spicules) and the Plakinidae (with simple spicules). In May 1990, an unusual sponge was found deep inside a submarine cave in the western Mediterranean Sea. On the basis of externally visible characters this sponge appeared indistinguishable from the common plakinid species Corticium candelabrum Schmidt, 1862. However, on closer examination in the laboratory the new sponge proved to be devoid of spicules. Therefore, despite great morphological similarities to C. candelabrum, the new sponge should, by taxonomic convention, have been placed in the Oscarellidae. On the basis of other criteria, the similarities to C. candelabrum were great and the new sponge was at first considered to be conspecific. Thus, the taxonomic position of the new sponge and its relationship to C. candelabrum are highly confusing. It could be an aspiculate morph of C. candelabrum, or a new and undescribed related species or, lacking spicules, it could justifiably be placed in a different family (Oscarellidae). The relationship of the new sponge to C. candelabrum and also to two species of Oscarella (Oscarellidae) was assessed by the use of enzyme electrophoresis to estimate genetic divergence between species. It was found that the new sponge was reproductively isolated from sympatric C. candelabrum, with 6 of 16 loci proving diagnostic. Thus it is clear that the new sponge belongs to a different biological species. Surprisingly it was also found that, although this new species was fairly closely related to C. candelabrum (level of genetic identity, I0.47), the two Oscarella species were similarly closely related to C. candelabrum (I0.31 to 0.41) and rather less closely to the new species (I0.17 to 0.28). Indeed from genetic identity estimates, O. tuberculata is more closely related to C. candelabrum than it is to O. lobularis. It is concluded that all homoscleromorph sponges should be placed in the single family Plakinidae.     

Evidence for a symbiosis between bacteria of the genus Rhodobacter and the marine sponge Halichondria panicea : harbor also for putatively toxic bacteria?

Halichondria panicea (Pallas) is a marine sponge, abundantly occurring in the Adriatic Sea, North Sea, and Baltic Sea. It was the aim of the present study to investigate if this sponge species harbors bacteria. Cross sections through H. panicea were taken and inspected by electron microscopy. The micrographs showed that this sponge species is colonized by bacteria in its mesohyl compartment. To identify the bacteria, polymerase chain reaction (PCR) analysis of the 16S rRNA gene segment, typical for bacteria, was performed. DNA was isolated from sponge material that had been collected near Rovinj (Adriatic Sea), Helgoland (North Sea), and Kiel (Baltic Sea) and was amplified with bacterial primers by PCR. The data gathered indicate that in all samples bacteria belonging to the genus Rhodobacter (Proteobacteria, subdivision α) are dominant, suggesting that these bacteria live in symbiotic relationship with the sponge. In addition, the results show that the different samples taken contain further bacterial species, some of them belonging to the same genus even though found in sponges from different locations. The possibility of the presence of toxic bacteria was supported by the finding that organic extracts prepared from sponge samples displayed toxicity, when analyzed in vitro using leukemia cells. Received: 7 March 1997 / Accepted: 2 October 1997     

Embryogenesis and acquisition of algal symbionts by planulae of Xenia umbellata (Octocorallia: Alcyonacea)

Early embryogenesis of the internally brooding soft coral Xenia umbellata and acquisition of algal symbionts in the course of its planular ontogenesis have been examined by scanning and transmission electron microscopy and by light microscopy. The endoderm of adult X. umbellata harbours symbionts mainly in the tentacles and in the peripheral solenia system. The colonies are gonochoric brooders. Algal symbionts were never found in the sperm sacs, and were only rarely found in the follicular tissue enclosing the oocytes. Fertilized eggs pass into endodermal brood pouches where embryogenesis occurs. Cleavage is holoblastic and leads to formation of a solid blastula. Algal symbionts are conspicuously embedded in the parental mesoglea that coats the young embryo, most probably transmitted by surface adherence. At a further stage, this integument disappears and the algae reside extracellularly among the cells of the newly-formed blastula. After subsequent cell proliferation developing planulae possess an inner mass of yolk-laden cells that contain numerous symbiotic algae. Gradually the yolk disintegrates, leaving a cavity enclosed by ectoderm, a thin mesoglea and an inner endoderm with intracellular symbionts. The mature planulae have already been provided with numerous intracellular symbionts by the time they are expelled from the brood pouches. The markedly early symbiont acquisition by the embryos of X. umbellata may help support their developmental requirements in the course of planular ontogenesis.     

Identification of the antifungal peptide-containing symbiont of the marine sponge Theonella swinhoei as a novel δ-proteobacterium, “Candidatus Entotheonella palauensis”

 The Palauan sponge Theonella swinhoei (class Demospongiae, order Lithistida, family Theonellidae) harbors filamentous bacterial symbionts that contain theopalauamide, an antifungal, bicyclic glycopeptide. In this study, the filamentous symbionts were shown to be novel bacteria belonging to the δ-subdivision of proteobacteria. The 16S rRNA gene sequence was determined using a combination of denaturing gradient-gel electrophoresis (DGGE) and specific polymerase chain-reaction (PCR) primers, and its source was confirmed by in situ hybridization. In a series of culture experiments, the filamentous bacteria were propagated in a mixed culture on agar plates. Related 16S rRNA gene sequences were isolated from related sponges with slightly different chemistry. The taxonomic status “Candidatus Entotheonella palauensis” is proposed for the theopalauamide-containing filamentous bacteria from T. swinhoei. Received: 12 June 1999 / Accepted: 22 January 2000     

Endofauna differences between two temperate marine sponges (Demospongiae; Haplosclerida; Chalinidae) from southwest Australia

The endofaunal assemblages associated with two species of sponge from the family Chalinidae (Haliclona sp. 1 and Haliclona sp. 2) were studied at four locations along the south west coast of Australia. The species have distinct morphologies and inhabit similar microhabitats; there is also considerable scientific interest in Haliclona sp. 1 (green Haliclona) due to the unique bioactive compound it produces. A total of 948 and 287 endofaunal individuals were found associated with 16 specimens of both the green Haliclona and Haliclona sp. 2 (brown Haliclona), respectively. Twenty-four endofaunal taxa were found (from mysid shrimps to teleost fish), with the brown Haliclona having a greater density of endofaunal species and individuals than the green Haliclona. The endofaunal assemblages of both species of sponge were significantly different, but only the endofaunal assemblage within the green Haliclona differed significantly among locations. Differences in the abundance and biomass of associated endofauna of each species of sponge can be related to differences in their morphologies, size and internal structure. In the green Haliclona, differences in endofaunal assemblages among locations are unlikely to be due to environmental influences as taxa discriminating each locations assemblage were common to both species of sponge. Numerous endofaunal individuals were found to be reproductively active, and it is clear that the species of sponge provide important habitats for their associated endofauna. This provision of habitat needs to be taken into account when harvesting green Haliclona biomass for supply of its target bioactive compound for further pharmaceutical development.     

Consistent association of crenarchaeal Archaea with sponges of the genus Axinella

The association of sponges with microorganisms has been accepted to be a common feature of the phylum Porifera. Herein we describe the association between filamentous Archaea and three Mediterranean species of sponges from the family Axinellidae (Porifera: Demospongiae). Axinella damicornis, A. verrucosa and Axinella sp. harbor a high concentration of filamentous Archaea in the collagen that surrounds the siliceous spicules that form their skeleton. These Archaea have been found in every axinellid specimen studied, regardless of their environment and collection time. The morphology of the filaments has been studied using transmission electron microscopy, and they all show similar characteristics. Their nature has been determined by in situ hybridization experiments and by PCR amplification and sequencing of their 16S DNA. Each sponge species contains a single filamentous archaeal phylotype. The Archaea of the three sponges are closely related to each other and to the marine "group 1" crenarchaeotes. Our findings suggest that this newly described association could be defined as a symbiosis, where biochemical and/or metabolic relationships between the sponge hosts and their symbionts remain to be determined.     

The importance of methane and thiosulfate in the metabolism of the bacterial symbionts of two deep-sea mussels

Undescribed hydrocarbon-seep mussels were collected from the Louisiana Slope, Gulf of Mexico, during March 1986, and the ultrastructure of their gills was examined and compared to Bathymodiolus thermophilus, a mussel collected from the deep-sea hydrothermal vents on the Galápagos Rift in March 1985. These closely related mytilids both contain abundant symbiotic bacteria in their gills. However, the bacteria from the two species are distinctly different in both morphology and biochemistry, and are housed differently within the gills of the two mussels. The symbionts from the seep mussel are larger than the symbionts from B. thermophilus and, unlike the latter, contain stacked intracytoplasmic membranes. In the seep mussel three or fewer symbionts appear to be contained in each host-cell vacuole, while in B. thermophilus there are often more than twenty bacteria visible in a single section through a vacuole. The methanotrophic nature of the seep-mussel symbionts was confirmed in ¹⁴C-methane uptake experiments by the appearance of label in both CO₂ and acid-stable, non-volatile, organic compounds after a 3 h incubation of isolated gill tissue. Furthermore, methane consumption was correlated with methanol dehydrogenase activity in isolated gill tissue. Activity of ribulose-1,5-biphosphate (RuBP) carboxylase and ¹⁴CO₂ assimilation studies indicate the presence of either a second type of symbiont or contaminating bacteria on the gills of freshly captured seep mussels. A reevaluation of the nutrition of the symbionts in B. thermophilus indicates that while the major symbiont is not a methanotroph, its status as a sulfur-oxidizing chemoautotroph, as has been suggested previously, is far from proven.     

Bacterial symbiosis in Northeast Pacific Vestimentifera: a TEM study

A number of new vestimentiferan species occur at northeast Pacific hydrothermal vent sites. The trophosome and bacterial symbionts of three species, collected from the Juan de Fuca and Explorer Ridges between 1984 and 1986, were studied by transmission electron microscopy (TEM). As in Riftia pachyptila, trophosome tissue is organised into lobules each having an axial blood vessel, and intracellular bacterial symbionts are contained in membrane vacuoles. The bacteria have many cytoplasmic inclusions including tubular membrane systems, glycogen-like particles and poly--hydroxybutyrate (PHB) or sulfur bodies. Glycogen production may be quantitatively important to both the symbionts and the host. Glycogen-like granules appear to first accumulate in the bacterial cells and then be released into the bacteriocyte cytoplasm as bacteria are degraded. Although various stages of bacterial growth and degradation are observed, data are insufficient to verify any across-lobule progression of these processes. Morphological comparison of the symbionts reveals that similar symbionts are found in different vestimentiferan species and that one to two bacterial types exist within single individuals. Two possible models of trophosome function and nutrient exchange are discussed.Deceased     

Biodiversity and rockfish recruitment in sponge gardens and bioherms of southern British Columbia,Canada

In the Strait of Georgia and Howe Sound, British Columbia, colonies of individual cloud sponges, growing on rock (known as sponge gardens) receive resource subsidies from the high biodiversity of epifauna on adjacent rock habitats. Bioherms are reefs of glass sponges living on layers of dead sponges. In the same area as the sponge gardens, newly discovered bioherms in Howe Sound, BC (49.34.67 N, 123.16.26 W) at depths of 28- to 35-m are constructed exclusively by Aphrocallistes vastus, the cloud sponge. The sponge gardens had much higher taxon richness than the bioherms. The sponge garden had 106 species from 10 phyla, whereas the bioherm had only 15 species from 5 phyla. For recruiting juvenile rockfish (quillback, Sebastes maliger), the food subsidy of sponge gardens appears to be missing on bioherms of cloud sponge, where biodiversity is relatively low. While adult and subadult rockfishes (S. maliger, S. ruberrimus, S. proriger, and S. elongatus) were present on bioherms, no evidence for nursery recruitment of inshore rockfishes to bioherms was observed, whereas the sponge gardens supported high densities of newly recruited S. maliger, perhaps owing to the combination of both refuge and feeding opportunities. These results indicate that sponge gardens form a habitat for early stages of inshore S. maliger, whereas A. vastus bioherms are associated only with older juvenile and adult rockfishes.     

The origin of the chemical profiles of fungal symbionts and their significance for nestmate recognition in <Emphasis Type="Italic">Acromyrmex</Emphasis> leaf-cutting ants

Cuticular hydrocarbon profiles are essential for nestmate recognition in insect societies, and quantitative variation in these recognition cues is both environmentally and genetically determined. Environmental cues are normally derived from food or nest material, but an exceptional situation may exist in the fungus-growing ants where the symbiotic fungus garden may be an independent source of recognition compounds. To investigate this hypothesis, we quantified the chemical profiles of the fungal symbionts of 18 sympatric colonies of Acromyrmex echinatior and Acromyrmex octospinosus and evaluated the quantitative variation of the 47 compounds in a multivariate analysis. Colony-specific chemical profiles of fungal symbionts were highly distinct and significantly different between the two ant species. We also estimated the relative genetic distances between the fungal symbionts using amplified fragment length polymorphism (AFLP) and correlated these with the overall (Mahalanobis) chemical distances between the colony-specific profiles. Despite the standardized laboratory conditions, the correlations were generally weak, but a statistically significant portion of the total variation in chemical profiles could be explained by genetic differences between the fungal symbionts. However, there was no significant effect of ant species in partial analyses because genetic differences between symbionts tend to coincide with being reared by different ant species. However, compound groups differed significantly with amides, aldehydes, and methyl esters contributing to the correlations, but acetates, alkanes, and formates being unrelated to genetic variation among symbionts. We show experimentally that workers that are previously exposed to and fed with the fungal symbiont of another colony are met with less aggression when they are later introduced into that colony. It appears, therefore, that fungus gardens are an independent and significant source of chemical compounds, potentially contributing a richer and more abundant blend of recognition cues to the colony “gestalt” than the innate chemical profile of the ants alone. Freddie-Jeanne Richard and Michael Poulsen contributed equally to this work.     

Genetic evidence for cryptic speciation in allopatric populations of two cosmopolitan species of the calcareous sponge genusClathrina

Many sponge species are considered to be cosmopolitan. However, the systematics of marine sponges are very difficult because of the paucity of taxonomically useful characters, and hence the apparently cosmopolitan nature of many species may be simply a consequence of this. In this paper, geographically distant populations of two pairs of cosmopolitan calcareous sponges of the genusClathrina were compared genetically.C. clathrus andC. cerebrum were collected by SCUBA diving between January and March 1989 from two localities: the Mediterranean Sea at La Vesse, near Marseille, Frances, at 9 to 12 m depth, and from the South West Atlantic at Arraial do Cabo, about 200 km east of Rio de Janeiro, at 2 to 10 m depth. Very high levels of gene divergence were found between the allegedly conspecific populations. The levels of genetic identity,I, observed are so low (I=0.128 and 0.287) that the populations clearly cannot be considered conspecific. New species names ofC. aurea sp. nov. andC. brasiliensis sp. nov. are therefore assigned to the southwest Atlantic counterparts ofC. clathrus andC. cerebrum, respectively. It is concluded that, at least for the species studied, and probably for many other species in taxonomically difficult groups, the actual distributions of single species may be far more geographically restricted than is generally assumed.