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Faecal pollution of water by bacteria has a negative effect on water quality and can pose a potential health hazard. Conventional surveillance of microbial water quality relies on the analysis of low-frequency spot samples and is thus likely to miss episodic or periodic pollution. This study aimed to investigate the potential of filter-feeding sponges for time-integrated biomonitoring of microbial water quality. Laboratory trials tested the effects of different ratios of bacterial abundance and the sequence of exposure on bacterial retention by the freshwater sponge Ephydatia fluviatilis (Linnaeus, 1759) to establish its potential to indicate bacterial exposure. Gemmule grown sponges were simultaneously exposed to Escherichia coli and Enterococcus faecalis but at different ratios (Trial 1) or individually exposed to each bacterial species but in different sequential order (Trial 2). The E. coli and E. faecalis retained in each sponge was quantified by culture on selective agars. Data analysis was conducted using the Kruskal-Wallis test and/or the Mann-Whitney U test to compare between the numbers of bacteria retained in each treatment. Additionally, the Wilcoxon matched-paired signed-rank test was used for comparison of the different bacterial abundances retained within each individual sponge. Sponges from all trials retained E. coli and E. faecalis in small numbers relative to the exposure (<0.05 % Trial 1 and <0.07 % Trial 2) but exhibited higher retention of E. coli. Higher abundance of either bacterial species resulted in significantly lower (P<0.005) retention of the same species within sponges (Trial 1). An initial exposure to E. coli resulted in significantly higher (P=0.040) retention of both bacterial species than when sponges were exposed to E. faecalis first (Trial 2).Bacterial retention by sponges was neither quantitatively representative of bacterial abundance in the ambient water nor the sequence of exposure. This implies either selective filtration or an attempt by sponges to prevent infection. However, freshwater sponges may still be useful in biomonitoring as qualitative time-integrated samplers of faecal indicator bacteria as they detect different bacteria present in the water even if their quantities cannot be estimated.

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Freshwater sponges (Spongillida: Demospongiae), including more than 240 described species, are globally distributed in continental waters (except for Antarctica), where they cover both natural and artificial surfaces. However, fragmentary studies have targeted their microbiome, making it difficult to test hypotheses about sponge-microbe specificity and metabolic relationships, along with the environmental factors playing key roles in structuring the associated microbial communities. To date, particular attention has been paid to sponges (family Lubomirskiidae) that are endemic to Lake Baikal. Few other freshwater sponge species (e.g., Ephydatia spp., Eunapius spp., and Spongilla lacustris), from lakes and rivers spanning from Europe to South and North America, have been targeted for microbiological studies. Representatives of the phyla Proteobacteria, Bacteroidetes, and Actinobacteria largely predominated, and high differences were reported between the microbiome of freshwater and marine sponges. Several bacterial strains isolated from freshwater sponges can produce bioactive compounds, mainly showing antibiotic activities, with potential application in biotechnology. Understanding the roles played by sponge microbiomes in freshwater ecosystems is still in its infancy and has yet to be clarified to disentangle the ecological and evolutionary significance of these largely under-investigated microbial communities. This review was aimed at providing the main available information on the composition and biotechnological potential of prokaryotic communities associated with healthy freshwater sponges, as a neglected component of the global sponge microbiome, to stimulate researchers interested in the field.

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Sponges (type Porifera) are multicellular organisms that give shelter to a variety of microorganisms: fungi, algae, archaea, bacteria, and viruses. The studies concerning the composition of viral communities in sponges have appeared rather recently, and the diversity and role of viruses in sponge holobionts remain largely undisclosed. In this study, we assessed the diversity of DNA viruses in the associated community of the Baikal endemic sponge, Baikalospongia bacillifera, using a metagenomic approach, and compared the virome data from samples of sponges and Baikal water (control sample). Significant differences in terms of taxonomy, putative host range of identified scaffolds, and functional annotation of predicted viral proteins were revealed in viromes of sponge B. bacillifera and the Baikal water. This is the evidence in favor of specificity of viral communities in sponges. The diversity shift of viral communities in a diseased specimen, in comparison with a visually healthy sponge, probably reflects the changes in the composition of microbial communities in affected sponges. We identified many viral genes encoding the proteins with metabolic functions; therefore, viruses in Baikal sponges regulate the number and diversity of their associated community, and also take a part in the vital activity of the holobiont, and this is especially significant in the case of damage (or disease) of these organisms in unfavorable conditions. When comparing the Baikal viromes with similar datasets of marine sponge (Ianthella basta), in addition to significant differences in the taxonomic and functional composition of viral communities, we revealed common scaffolds/virotypes in the cross-assembly of reads, which may indicate the presence of some closely related sponge-specific viruses in marine and freshwater sponges.

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This paper reports the discovery of a small population of sponges in the Pigeon River of eastern Tennessee, USA, which were morphologically distinct from Spongillida of North America. A morphological comparative analysis resulted in the first Nearctic record of the genus Heterorotula with the description of a new species Heterorotulalucasi sp. nov. diverging from all other known species by its unique combinations of diagnostic morphotraits of spicules and gemmules. The new record enlarges the geographic range of the genus which has been known until now only from Australia, New Zealand, New Caledonia, Japan (as an alien species), and from subequatorial Brazil (as subfossil remains). The discovery of a biogeographic enclave of Heterorotula in the southeastern United States contributes to the understanding of Porifera inland water biodiversity, biogeographic patterns, and adaptive morphotraits in the Nearctic and globally. Data confirm that the Appalachian region (Ordovician-Permian origin) of Tennessee and, in general, of North America have high levels of diversity and endemicity.

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In many freshwater habitats, green algae form intracellular symbioses with a variety of heterotrophic host taxa including several species of freshwater sponge. These sponges perform important ecological roles in their habitats, and the poriferan:green algae partnerships offers unique opportunities to study the evolutionary origins and ecological persistence of endosymbioses. We examined the association between Ephydatia muelleri and its chlorophyte partner to identify features of host cellular and genetic responses to the presence of intracellular algal partners. Chlorella-like green algal symbionts were isolated from field-collected adult E. muelleri tissue harboring algae. The sponge-derived algae were successfully cultured and subsequently used to reinfect aposymbiotic E. muelleri tissue. We used confocal microscopy to follow the fate of the sponge-derived algae after inoculating algae-free E. muelleri grown from gemmules to show temporal patterns of symbiont location within host tissue. We also infected aposymbiotic E. muelleri with sponge-derived algae, and performed RNASeq to study differential expression patterns in the host relative to symbiotic states. We compare and contrast our findings with work in other systems (e.g., endosymbiotic Hydra) to explore possible conserved evolutionary pathways that may lead to stable mutualistic endosymbioses. Our work demonstrates that freshwater sponges offer many tractable qualities to study features of intracellular occupancy and thus meet criteria desired for a model system.

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Associations of freshwater sponges with coccoid green algae have been known for a long time. Two types of coccoid green algae, which are commonly assigned as zoochlorellae, are recognized by morphology: small coccoids (< 3 µm) without pyrenoids and larger Chlorella-like algae (4-6 µm) with pyrenoids. Despite their wide distribution in some freshwater sponges, these green algae were never studied using a combined analysis of morphology and molecular phylogeny. We investigated several endosymbiotic strains isolated from different Spongilla species, which were available in culture collections. Phylogenetic analyses of SSU and ITS rDNA sequences revealed that the strain SAG 211-40a is a member of the Chlorellaceae and represents a new species of the newly erected genus Lewiniosphaera, L symbiontica. The phylogenetic position was confirmed by morphology and ITS-2 barcode. The endosymbionts without pyrenoid were identified as Choricystis parasitica by morphology and phylogenetic analyses. The comparison with free-living strains revealed the recognition of two new Choricystis species, C. krienitzii and C. limnetica, which were confirmed by molecular signatures in V9 region of SSU rDNA and ITS-2 barcode.

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Sponges are ecologically important components of marine and freshwater benthic environments; these holobionts contain a variety of microorganisms and viruses. For the metagenomic characterization of potential taxonomic and functional diversity of sponge-associated dsDNA viruses, we surveyed two samples of Baikal endemic sponge Baikalospongia bacillifera (diseased and visually healthy). In total, after quality processing, we have obtained 3 375 063 and 4 063 311 reads; of these 97 557 and 88 517 sequences, accounting for ca. 2.9 and 2.2% of datasets, have been identified as viral. We have revealed approximately 28 viral families, among which the bacteriophages of the Myoviridae, Siphoviridae and Podoviridae families, as well as the viruses of the Phycodnaviridae and Poxviridae families, dominated in the samples. Analysis of viral sequences using the COG database has indicated 22 functional categories of proteins. Viral communities of visually healthy and diseased Baikal sponges were significantly different. The metagenome sequence data were deposited to NCBI SRA as BioProject PRJNA577390.

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We report here the first record of five genera of Spongillida (Porifera, Demospongiae) in both lentic and lotic water of Madagascar and the discovery of a new species of the genus Corvospongilla from the Betsiboka River hydrographic basin. Corvospongilla lemuriensis sp. n. diverges from all the other known species of the genus in its unique combination of diagnostic morphotraits (a) sessile gemmules with chambered pneumatic layer within tri-layered theca and enveloped by spicular cage of dominant acanthostrongyles, (b) dominant acanthostrongyles and less frequent acanthoxeas-strongyloxeas as megascleres and gemmuloscleres. The new species is characterised by the exclusive presence of a well-developed pneumatic layer in sessile gemmules not described for the genus until now. The new record confirms a Gondwanian track for the genus Corvospongilla and increases the knowledge on Spongillida historical biogeography. In synthesis the present discovery of Corvospongilla in the Malagasy biogeographic province (a) contributes to the assessment of Afrotropical biodiversity, (b) enlarges the geographic range of the genus reducing its disjunct distribution, and (c) focus on the evolutionary history of adaptive morphofunctional traits of resting bodies and life cycle timing in ephemeral water of the south-eastern Austral hemisphere.

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Successful dispersal of freshwater sponges depends on the formation of dormant sponge bodies (gemmules) under adverse conditions. Gemmule formation allows the sponge to overcome critical environmental conditions, for example, desiccation or freezing, and to re-establish as a fully developed sponge when conditions are more favorable. A key process in sponge development from hatched gemmules is the construction of the silica skeleton. Silica spicules form the structural support for the three-dimensional filtration system the sponge uses to filter food particles from ambient water. We studied the effect of different hypergravity forces (1, 2.5, 5, 10, and 20 × g for 48 h)-as measure for environmental stress-on the ability of developing sponges to set-up their spiculous skeleton. Additionally, we assessed whether the addition of nutrients (i.e., dissolved 13C- and 15N-labeled amino acids) compensates for this stress. Our results show that freshwater sponges can withstand prolonged periods of hypergravity exposure and successfully set-up their skeleton, even after 48 h under 20 × g. Developing sponges were found to take up and assimilate dissolved food before forming a functional filtering system. However, fed and non-fed sponges showed no differences in skeleton formation and relative surface area growth, suggesting that the gemmules' intrinsic energy fulfills the processes of skeleton construction. Additionally, non-fed sponges formed oscula significantly more often than fed sponges, especially under higher g-forces. This suggests that the eventual formation of a filtration system might be stimulated by food deprivation and environmentally stressful conditions. These findings indicate that the process of spiculous skeleton formation is energy-efficient and highly resilient. The uptake of dissolved food substances by freshwater sponges may contribute to the cycling of dissolved organic matter in freshwater ecosystems where sponges are abundant.

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African sponges, particularly freshwater sponges, are understudied relative to demosponges in most other geographical regions. Freshwater sponges (Spongillida) likely share a common ancestor; however, their evolutionary history, particularly during their radiation into endemic and allegedly cosmopolitan groups, is unclear. Freshwater sponges of at least 58 species of 17 genera and four families are described from Central and Eastern Africa, but the diversity is underestimated due to limited distinguishable morphological features. The discovery of additional cryptic species is very likely with the use of molecular techniques such as DNA barcoding. The Royal Museum of Central Africa (MRAC, Tervuren, Belgium) hosts one of the largest collections of (Central) African freshwater sponge type material. Type specimens in theory constitute ideal targets for molecular taxonomy; however, the success is frequently hampered by DNA degradation and deamination, which are a consequence of suboptimal preservation techniques. Therefore, we genotyped African demosponge holotype material of the MRAC with specific short primers suitable for degenerated tissue and compare the results with the current, morphology-based classification. Our results demonstrate the utility of minimalistic barcodes for identification of sponges, potentially enabling efficient identification of individuals in taxonomic or metabarcoding studies, and highlight inconsistencies in the current freshwater sponge classification.

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Anheteromeyenia Schröder, 1927 (Spongillidae) was erected to comprise species of Heteromeyenia Potts, 1881 without microscleres. Until now, only four species of Anheteromeyenia are known, three from the Neotropical Region (A. cheguevarai, A. ornata and A. vitrea) and A. argyrosperma from the Nearctic Region. In the present study, we describe a new species of Anheteromeyenia from the Neotropical Region, which differs from other congeners in having only one category of megascleres, exclusively smooth oxeas. An identification key to species of Anheteromeyenia and an emended diagnosis of the genus are provided.

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The psbA gene, which encodes a major photosystem II protein (protein II or D1), is a marker for the presence of phototrophic organisms in water communities. We have pioneered the use of this marker for studying the diversity of phototrophic microflora of freshwater invertebrates. The object of the study is the microbial associations accompanying the endemic Baikal sponge Baikalospongia intermedia and the surrounding aquatic microbial community. Analysis of the psbA gene sequences in the examined microbiomes demonstrates the presence of various phototrophic groups, such as Cyanobacteria, Chlorophyta, Heterokonta, Haptophyta, and Ochrophyta algae, as well as cyanophages. A total of 35 unique psbA gene sequences have been distinguished in the microbial communities of the endemic sponge B. intermedia and 32 unique sequences in the water community surrounding the sponge. These data demonstrate the involvement of sponge symbiotic communities in the accumulation of primary production and carbon cycle in the Lake Baikal ecosystem.

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Sponges are primitive metazoans that are known to harbour diverse and abundant microbes. All over the world attempts are being made to exploit these microbes for their biotechnological potential to produce, bioactive compounds and antimicrobial peptides. However, the majority of the studies are focussed on the marine sponges and studies on the freshwater sponges have been neglected so far. To increase our understanding of the microbial community structure of freshwater sponges, microbiota of two fresh water sponges namely, Eunapius carteri and Corvospongilla lapidosa is explored for the first time using Next Generation Sequencing (NGS) technology. Overall the microbial composition of these sponges comprises of 14 phyla and on an average, more than 2900 OTUs were obtained from C. lapidosa while E. carteri showed 980 OTUs which is higher than OTUs obtained in the marine sponges. Thus, our study showed that, fresh water sponges also posses highly diverse microbial community than previously thought and it is distinct from the marine sponge microbiota. The present study also revealed that microbial community structure of both the sponges is significantly different from each other and their respective water samples. In the present study, we have detected many bacterial lineages belonging to Firmicutes, Actinobacteria, Proteobacteria, Planctomycetes, etc. that are known to produce compounds of biotechnological importance. Overall, this study gives insight into the microbial composition of the freshwater sponges which is highly diverse and needs to be studied further to exploit their biotechnological capabilities.

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The freshwater sponge species Ephydatia cf. facunda Weltner, 1895 (Spongillida, Spongillidae) is reported for the first time as a fossil from middle Eocene lake sediments of the Giraffe kimberlite maar in northern Canada. The sponge is represented by birotule gemmuloscleres as well as oxea megascleres. Today, E. facunda inhabits warm-water bodies, so its presence in the Giraffe locality provides evidence of a warm climate at high latitudes during the middle Eocene. The morphological similarity of the birotules to modern conspecific forms suggests protracted morphological stasis, comparable to that reported for other siliceous microfossils from the same locality.

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Freshwater sponges are abundant in the Amazon region and they have been known to cause dermatitis (acute inflammation) since the beginning of the 20th century. To determine whether additional constituents, besides their body spicules, cause dermatological reactions in humans, an experimental study was developed and carried out using mice and Drulia uruguayensis prepared in three different forms: intact sponges (IS), macerated sponges (MS) or isolated spicules - megascleres (ISM). The cells most commonly involved in inflammatory reactions (mast cells, eosinophils and neutrophils), as well as intraepithelial lymphocytes and degranulated mast cells, were counted so that they could be used as parameters to determine which of the sponge preparations induced the greatest reaction. The effects of the sponge on the skin were then determined by histological analysis. The results obtained showed that IS caused the greatest inflammatory reaction (p = 0.000005), activating mainly mast cells (p = 0.0018). The histopathological analysis revealed a slight loss of continuity of the epidermis when ISM or IS were applied. These findings allow us to conclude that a structurally intact sponge can cause a greater inflammatory reaction in the first contact because of its ability to perforate the skin and allow inflammatory agents to enter. Other proteins present in dried sponge bodies could induce allergic but not toxic responses (in contact with the entire sponge, a large number of pharmacologically inert proteins may be introduced, with a potential allergen).

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