Phylogenetic relationships of freshwater sponges (Porifera, Spongillina) inferred from analyses of 18S rDNA, COI mtDNA, and ITS2 rDNA sequences

The phylogenetic relationships of nine species of freshwater sponges, representing the families Spongillidae, Lubomirskiidae, and Metaniidae, were inferred from analyses of 18S rDNA, cytochrome oxidase subunit I (COI) mtDNA, and internal transcribed spacer 2 (ITS2) rDNA sequences. These species form a strongly supported monophyletic group within the Demospongiae, with the lithistid Vetulina stalactites as the sister taxon. Within the freshwater sponge clade, the basal taxon is not resolved. Depending upon the method of analysis and sequence, the metaniid species, Corvomeyenia sp., or the spongillid species, Trochospongilla pennsylvanica , emerges as the basal species. Among the remaining freshwater sponge species, the spongillids, Spongilla lacustris and Eunapius fragilis , form a sister group to a clade comprised of the spongillid species, Clypeatula cooperensis , Ephydatia fluviatilis , and Ephydatia muelleri , and the lubomirskiid species, Baikalospongia bacillifera and Lubomisrkia baicalensis . C. cooperensis is the sister taxon of E. fluvialitis , and E. muelleri is the sister taxon of ( B. bacillifera + L. baicalensis ). The family Spongillidae and the genus Ephydatia are thus paraphyletic with respect to the lubomirskiid species; Ephydatia is also paraphyletic to C. cooperensis . We suggest that C. cooperensis be transferred to the genus Ephydatia and that the family Lubomirskiidae be subsumed into the Spongillidae.     

Distribution and ecology of freshwater sponges in Connecticut

A survey of Connecticut lakes and rivers revealed the presence of 7 species of freshwater sponge: Spongilla lacustris, Ephydatia muelleri, Eunapius fragilis, Anheteromeyenia ryderi, A. argyrosperma, Corvomeyenia carolinensis, and Corvospongilla novaeterrae in order of decreasing frequency of occurrence. Corvomeyenia carolinensis has not been reported previously beyond its type locality in South Carolina. In addition, microscleres of Spongilla lacustris, Anheteromeyenia-like megascleres, Ephydatia muelleri-like megascleres, and smooth megascleres (amphioxeas), which could not be assigned to a particular species, were found in surface sediments from lake cores. Spongilla lacustris inhabiting small rivers produced brown, thick-capsuled gemmules during the summer and yellow, thin-capsuled gemmules during the fall. The thick-capsuled gemmules, but not the thin-capsuled gemmules, are tolerant of desiccation; and populations of Spongilla lacustris and Ephydatia muelleri survived severe drying of their habitats during the summer.     

Life-cycles of lotic populations of Spongilla lacustris and Eunapius fragilis (Porifera: Spongillidae)

SUMMARY. The life-cycles of green and white morphs of the freshwater sponge Spongilla lacustris were examined in the light of past evidence that zoochlorellae may augment their sponge host's nutrition. Field collections from a lotic population of S. lacustris were supplemented by laboratory experiments on gemmule hatching and gemmule size. Both white and green S. lacustris produced sperm for a 6-week period in 1976 starting in the middle of May. Out of thirty white and thirty green sponges examined during this period, twenty white and ten green sponges contained sperm. Sperm production in both morphs was limited primarily to the basal 3.18mm of sponge tissue, and the density of sperm packets in the two morphs was the same. Out of 180 white and green sponges examined in 1976, only four eggs, no embryos, and no larvae were observed. White sponges gemmulated a week or two earlier, and produced smaller gemmules which were more uniform in size than those of green sponges. White and green gemmules hatched synchronously in the spring. In 1977 one female and numerous male specimens of S. lacustris , and numerous females but no males of another sponge, Eunapius fragilis , were found. The life-cycles are discussed in the light of other recent studies on freshwater sponges.     

Freshwater sponge silicateins: comparison of sequences and exon-intron structure of genes

Siliceous sponge spicules contain silicateins--proteins taking part in biogenic silica precipitation and determination of the spicule morphological features. The exon-intron structure of four silicatein-alpha isoforms: -alpha1,-alpha2, -alpha3 and -alpha4 from endemic baikalian sponge Lubomirskia baicalensis was studied. For eight sponge species, including both cosmopolitan (Spongilla lacustris, Ephydatia muelleri, E. fluviatilis) and Baikal endemic (L. baicalensis, L. incrustans, Baikalospongia intermedia, B. fungiformis, Sw. papyracea) species, seventeen gene fragment sequences of different silicatein isoforms were determined. It was shown that cosmopolitan and endemic Baikalian sponges differ from each other by gene structure (have different length ofintrons). Among Baikalian sponges silicatein-alpha1 has the most variable intron length, and silicatein-alpha4 is the most conservative. Phylogenetic analysis of amino-acid silicatein sequences allow identify different silicatein isoforms, which authentically differ form four clusters on phylogenetic tree. Phylogenetic analysis of exon-intron sequences gives the possibility to separate different sponge species in the clusters.     

Phylogenetic position of sponges from Chagytaĭ and Tore-Khol' lakes

Morphological and molecular genetic data for freshwater sponges from the lakes of Tuva Depression, Baikalospongia dzhegatajensis (Rezvo, 1936), forms Dzh05 and Dzh06, from Chagatai Lake, as well as forms TKhl and TKh2, from the Lake Tore-Khol, were obtained and examined. In the sponges examined, which on phylogenetic tree clustered together with the Ephydatia fluviatilis (Linneaus, 1758) sponge from the family Spongillidae, the ITS rDNA regions were sequenced. Comparison of highly variable interal spacer regions of the mitochondrial genome was performed using corresponding sequences of three sponges from the family Spongillidae (E. fluviatilis, E. muelleri and Spongilla lacustris), sponges from the Chagatai and Tore-Khol lakes (Dzh06 and TKh2) with an unknown status, and sponges from the Baikalian family Lubomirskiidae. Minimum genetic differences were observed between E. fluviatilis, Dzh06, and TKh2 (from 0.003 to 0.01% of nucleotide substitutions), while maximum differences were found between the species of Lubomirskiidae and Spongillidae (from 0.928 to 2.06%). The data obtained indicated that sponges from Chagatai and Tore-Khol lakes were most close to E. fluviatilis.     

Spatial and temporal distribution of freshwater sponges in Connecticut lakes based upon analysis of siliceous spicules in dated sediment cores

A survey of 28 Connecticut water bodies for spicular remainsoffreshwater sponges preserved in lacustrine sediments revealed14species representing 10 genera. Sediments were examined atfivelevels (ca 1991, ca 1970, ca 1934, ca 1890, ca 1875)todocument changes in species composition, distribution, andabundance. Five species were recorded statewide in all fivegeologic regions: Spongilla lacustris, Heteromeyeniatubisperma, Anheteromeyenia ryderi, Eunapius fragilis, andEphydatia muelleri. Spongilla lacustris was the mostcommonand widely distributed species. Factors that may havecontributedto the observed distribution of the spongillids in Connecticutareregional differences in lake water chemistries, individualspeciestolerances, and differences in extent of dispersal of variousspecies. Most of the more abundant species are widelydistributedwith respect to water quality parameters. Anheteromeyeniaargyrosperma, Duosclera mackayi and Trochospongillapennsylvanica appear to be restricted to waters exhibitingthelower ranges for conductivity, alkalinity and calcium content.Ingeneral, a greater number of spicules was found in bottomsedimentsof the surveyed lakes as compared with the surface sediments.Oftenthis was correlated with increased sediment accumulation ratesinmore recent periods. When this fact is taken into account, itappears that populations of a number of sponge species haveremained relatively constant in many of the lakes over thepastcentury. However, in some lakes exhibiting lower or onlyslightlyhigher sediment accumulation rates at the surface, fewerspiculeswere also found in more recent sediments, suggesting an actualdecline in sponge population sizes. Changes in abundance overthelast 100 years may be related to changes in land use practicesinsurrounding watersheds and the associated changes in waterchemistries.     

Freshwater sponge silicateins: Comparison of gene sequences and exon-intron structure

Silicateins found in spicules of siliceous sponges are proteins that take part in biogenic silica precipitation and determine the morphological features of spicules. The exon-intron structure of the genes encoding four silicatein-α isoforms (−α1, −α2, −α3, and −α4) from an endemic Baikalian sponge Lubomirskia baicalensis was studied. For eight sponge species, including both cosmopolitan (Spongilla lacustris, Ephydatia muelleri, E. fluviatilis) and endemic Baikalian (L. baicalensis, L. incrustans, Baikalospongia intermedia, B. fungiformis, Sw. papyracea) species, seventeen partial sequences of different silicatein isoform genes were determined. It was shown that cosmopolitan and endemic Baikalian sponges differ from each other in gene structure, in particular, in intron length. Among Baikalian sponges, silicatein-α1 genes had the highest variation of intron length, and silicatein-α4 genes were the most conservative. A phylogenetic analysis based on amino acid sequences of different silicatein isoforms identified four distinct clusters within the freshwater sponge clade. An analysis based on exon-intron gene sequences enables discrimination between different sponge species within the clusters.     

Cellular src gene product detected in the freshwater sponge Spongilla lacustris.

Serum from Rous sarcoma virus tumor-bearing rabbits immunoprecipitated from extracts of the freshwater sponge Spongilla lacustris a tyrosine-specific protein kinase with characteristics similar to the chicken pp60c-src kinase activity. An immune competition assay confirmed the relationship between the protein from sponges and viral pp60v-src.     

Mesoscale investigations based on microsatellite analysis of the freshwater sponge <Emphasis Type="Italic">Ephydatia fluviatilis</Emphasis> in the River-Sieg system (Germany) reveal a genetic divergence

Freshwater sponges play a major role in freshwater ecological system as important filter-feeding organisms and bioindicators. There are only few data about their ecological diversity and population genetic structure available, though a deeper knowledge is needed to propose proper conservation and effective management. The aim of this study was to assess data on distribution patterns of freshwater sponges to study the connectivity of genotypes of Ephydatia fluviatilis in a river system. We sampled specimens from River-Sieg system (River Agger and River Sieg, Germany). We hypothesized that strong anthropogenic influence would cause a uniform distribution of population structures. The genetic structure of E. fluviatilis populations was analysed with a set of eleven microsatellite loci from seven locations in River-Sieg system. Besides of E. fluviatilis, three other species co-occurred (Ephydatia mülleri, Spongilla lacustris, Eunapius fragilis). In contrast to our hypothesis, we observed an overall correlation between genetic and geographic distances among populations of this sessile species, which follows a clear isolation-by-distance pattern. A significant microsatellite polymorphism and high levels of genetic divergence between populations (F_ST) in upstream reaches were present. These results will provide important information for conservation management of populations with limited dispersal ability in connected river systems.     

Sponge cytogenetics - mitotic chromosomes of ten species of freshwater sponge

Porifera (sponges) are the most basal phylum of extant metazoans. To gain insight into sponge genome construction, cytogenetic analysis was performed for ten freshwater sponge species of six genera, using conventional Giemsa staining, chromosome banding, and fluorescence in-situ hybridization. The karyotypes were very similar among the ten species, exhibiting a diploid chromosome number of 2n=46 or 48, and usually consisted of microchromosomes with one or two pairs of large chromosomes. The 18S-28S rRNA genes were localized to a single pair of microchromosomes in two Ephydatia species. Hybridization signals of the telomere (TTAGGG)n sequences were observed at the ends of metaphase chromosomes. The genome sizes of Ephydatia fluviatilis and Ephydatia muelleri were estimated by flow cytometric analysis as about 0.7 pg per diploid complement. These freshwater sponge species appear to represent a fairly homogeneous group with respect to karyotypes.     

Freshwater sponge (Porifera: Spongillidae) distribution across a landscape: Environmental tolerances,habitats, and morphological variation

Freshwater sponges are important to ecosystem functioning; however, information about their biogeography and interspecific variation is fragmentary, limiting our ability to assess their role. Although the specific epithets of two common species suggest that sponges found in lentic habitats are Spongilla lacustris, and those found in lotic habitats are Ephydatia fluviatilis, the number of sponge species in the UK is unresolved. We sampled sponges in a variety of habitats and used both morphological and molecular (D3 domain of 28S rDNA) methods to identify six species, including the first record of Trochospongilla horrida. We contrasted species in terms of their environmental tolerances, habitats, and variation, and we expanded on the limited information available about the geographic distributions of these sponges. In our study, most sponge species colonized a variety of substrates, but exhibited different distributions. The most widespread sponge, S. lacustris, was present at lower mean water temperatures and was more often located above a latitude of 55°N. Ephydatia fluviatilis was the most common species in rivers, but was also located in lentic habitats. Salinity in anthropogenic habitats was not a significant factor for the presence of E. fluviatilis or the more patchily distributed species Eunapius fragilis. Instead, these species occurred more frequently at sites with negative oxidation–reduction potential. Sponge biodiversity may be affected by substrate availability in anthropogenic habitats, invasive species, and improved ability to recognize sponge taxa. Crucially, we provide foundation data as a prerequisite for future ecological evaluation.     

Phylogenetic analysis of freshwater sponges provide evidence for endemism and radiation in ancient lakes

Morphologic and phylogenetic analysis of freshwater sponges endemic to lakes in Central Sulawesi, Siberia and South-East Europe is presented. We also analyzed several cosmopolitan sponge species from Eurasia and North America and included sponge sequences from public databases. In agreement with previous reports [Addis, J.S., Peterson, K.J., 2005. Phylogenetic relationships of freshwater sponges (Porifera, Spongillina) inferred from analyses of 18S rDNA, COI mtDNA, and ITS2 rDNA sequences. Zool. Scr. 34, 549-557], the metaniid sponge Corvomeyenia sp. was the most deeply branching species within a monophyletic lineage of the suborder Spongillina. Pachydictyum globosum (Malawispongiidae) and Nudospongilla vasta (Spongillidae), two morphologically quite distinct species from Sulawesi were found in a joint clade with Trochospongilla (Spongillidae) rendering Trochospongilla paraphyletic. Furthermore, Ochridaspongia sp., another Malawispongiidae, clustered far away from that clade, together with Ephydatia fluviatilis, making the latter family polyphyletic. The Lubomirskiidae endemic to Lake Baikal, Lubomirskia abietina, Baikalospongia bacillifera, B. intermedia, and Swartschewskia papyracea formed a well-supported clade that was most closely linked to the genus Ephydatia (99.9% identity over a total length of 2169 concatenated nucleotide positions). Our study indicates the frequent and independent origin of sponge species endemic to different freshwater ecosystems from a few cosmopolitan founder species. The highly specific primer sets newly developed here facilitate work on the molecular phylogeny and DNA barcoding of sponges.     

Small inverted repeats drive mitochondrial genome evolution in Lake Baikal sponges

Demosponges, the largest and most diverse class in the phylum Porifera, possess mitochondrial DNA (mtDNA) markedly different from that in other animals. Although several studies investigated evolution of demosponge mtDNA among major lineages of the group, the changes within these groups remain largely unexplored. Recently we determined mitochondrial genomic sequence of the Lake Baikal sponge Lubomirskia baicalensis and described proliferation of small inverted repeats (hairpins) that occurred in it since the divergence between L. baicalensis and the most closely related cosmopolitan freshwater sponge Ephydatia muelleri. Here we report mitochondrial genomes of three additional species of Lake Baikal sponges: Swartschewskia papyracea, Rezinkovia echinata and Baikalospongia intermedia morpha profundalis (Demospongiae, Haplosclerida, Lubomirskiidae) and from a more distantly related freshwater sponge Corvomeyenia sp. (Demospongiae, Haplosclerida, Metaniidae). We use these additional sequences to explore mtDNA evolution in Baikalian sponges, paying particular attention to the variation in the rates of nucleotide substitutions and the distribution of hairpins, abundant in these genomes. We show that most of the changes in Lubomirskiidae mitochondrial genomes are due to insertion/deletion/duplication of these elements rather than single nucleotide substitutions. Thus inverted repeats can act as an important force in evolution of mitochondrial genome architecture and be a valuable marker for population- and species-level studies in this group. In addition, we infer (((Rezinkovia+Lubomirskia)+Swartschewskia)+Baikalospongia) phylogeny for the family Lubomirskiidae based on the analysis of mitochondrial coding sequences from freshwater sponges.     

The mitochondrial genome of stygobitic sponge Eunapius subterraneus: mtDNA is highly conserved in freshwater sponges

The complete mitochondrial DNA (mtDNA) genome of the Eunapius subterraneus (Porifera, Demospongiae), a unique stygobitic sponge, was analyzed and compared with previously published mitochondrial genomes from this group. The 24,850 bp long mtDNA genome is circular with the same gene composition as found in other metazoans. Intergenic regions (IGRs) comprise 24.7% of mtDNA and are abundant with direct and inverted repeats and palindromic elements as well as with open reading fames (ORFs) whose distribution and homology was compared with other available mt genomes with a special focus on freshwater sponges. Phylogenetic analyses based on concatenated amino acid sequences from 12 mt protein genes placed E. subterraneus in a well-supported monophyletic clade with the freshwater sponges, Ephydatia muelleri and Lubomirskia baicalensis. Our study showed high homology of mtDNA genomes among freshwater sponges, implying their recent split.     

Insights into the evolution of freshwater sponges (Porifera: Demospongiae: Spongillina): Barcoding and phylogenetic data from Lake Tanganyika endemics indicate multiple invasions and unsettle existing taxonomy

Sponges are a conspicuous element in many benthic habitats including in Africa's oldest, deepest lake, Lake Tanganyika. Despite their prevalence and pivotal ecological role as filter feeders, knowledge of the evolutionary history of sponges is in its infancy. Here, we provide the first molecular analysis targeting the evolution of sponges from Lake Tanganyika. Independent markers indicate the occurrence of several colonisation events which have shaped the current Tanganyikan lacustrine sponge biodiversity. This is in contrast to a range of previously studied organisms that have diversified within the lake from single lineages. Our tree reconstructions indicate the presence of two genera, Oncosclera and Eunapius, which are globally distributed. Therefore, we reject the hypothesis of monophyly for the sponges from Lake Tanganyika and challenge existing higher taxonomic structure for freshwater sponges.     

Molecular phylogeny of the freshwater sponges in Lake Baikal

The phylogenetic relationship of the freshwater sponges (Porifera) in Lake Baikal is not well understood. A polyphyletic and/or monophyletic origin have been proposed. The (endemic) Baikalian sponges have been subdivided into two families: endemic Lubomirskiidae and cosmopolitan Spongillidae. In the present study, two new approaches have been made to resolve the phylogenetic relationship of Baikalian sponges; analysis of (1) nucleotide sequences from one mitochondrial gene, the cytochrome oxidase subunit I (COI) and of (2) one selected intron from the tubulin gene. Specimens from the following endemic Baikalian sponge species have been studied; Lubomirskia baicalensis , Baikalospongia intermedia, Baikalospongia recta , Baikalospongia bacillifera and Swartschewskia papyracea . They are all grouped to the family of Lubomirskiidae. Sequence comparisons were performed with the ubiquitously distributed freshwater sponge Spongilla lacustris (family Spongillidae) as well as with one marine sponge, Suberites domuncula . A sequence comparison * * The sequences reported here are being deposited in the EMBL data base. of the mitochondrial COI gene revealed a monophyletic grouping of the endemic Baikalian sponges with S. lacustris as the most related species to the common ancestor. The sequences of the COI gene from B. recta , B. intermedia , B. bacillifera and L. baicalensis were found to be identical and separated from those of S. lacustris and S. papyracea . In a second approach, the exon/intron sequences framing the intron‐2 of the sponge tubulin gene were chosen for the phylogenetic analysis. The intron sequences were aligned and used for construction of a phylogenetic tree. This analysis revealed again a monophyletic grouping with S. lacustris as the closest related species to the common ancestor. It is concluded that the Baikalian sponges, which have been studied here, are of monophyletic origin. Furthermore, the data suggest that the endemic species S. papyracea is the phylogenetically oldest, extant, endemic Baikalian sponge species.     

Mitochondrial genomes of the genus Ephydatia Lamouroux, 1816: can palindromic elements be used in species-level studies?

Poriferan mitochondrial DNA (mtDNA), especially large intergenic regions, is a target for the insertion of repetitive hairpin-forming elements. These elements are responsible for the large mt genome size differences observed even among closely related sponge taxa. In this study, we present the new, nearly complete, mt genome sequence of Ephydatia fluviatilis and compare it with previously published mt genomes of freshwater sponges. Special emphasis was placed on comparison with the closely related species Ephydatia muelleri, thereby comparing the only two species of the genus Ephydatia on the western Balkan Peninsula. In particular, we analyzed repetitive palindromic elements within the mitochondrial intergenic regions. The genomic distribution of these repetitive elements was analyzed and their potential role in the evolution of mt genomes discussed. We show here that palindromic elements are widespread through the whole mt genome, including the protein coding genes, thus introducing genetic variability into mt genomes.     

Chlorophyll and Primary Productivity of Some Green,Freshwater Sponges

Relationships between ash-free dry weight, viable chlorophyll a and pheopigments a were examined by linear regression analysis for non-gemmulating and gemmulated Spongilla lacustris and for an unidentified, non-gemmulating species. The primary productivity of S. lacustris was determined by both oxygen and ¹⁴C techniques and expressed on a biomass, chlorophyll a, and area basis, and the photosynthetic efficiency was calculated. The proportion of algal photosynthate excreted by the sponge was investigated, and the transfer of algal photosynthate to the sponge was examined by autoradiography.     

A continuous-flow system for growing fresh-water sponges in the laboratory

Two fresh-water sponge species, Ephydatia fluviatilis and Spongilla alba, were grown from gemmules in the laboratory. A system incorporating a continuous flow of filtered habitat water and live bacteria from a chemostat culture as a food source were used. Experiments with this system demonstrated a relationship between the concentration of bacteria and sponge growth rate. Because the continuous flow of water eliminates the effects of substances released by sponges and growth rate can be predicted for a given bacterial concentration, this system permits experimental studies which were not feasible in the past.