Identification Of Sponges Research Articles

CHECKLIST OF MARINE SPONGES OFF BIDONG ARCHIPELAGOS BASED ON COI GENE SEQUENCES

Sponges are sessile marine animals under the phylum Porifera. Conventionally, the identification of sponges is based on two criteria: Morphology (shape, colour, surface characteristics and consistency) and macro-morphology (spicules, sponging fibres and their arrangement). This identification technique is sometimes unreliable due to the plasticity of sponges’ characteristics, which causes misidentification. We utilized universal barcode COI primers to identify sponges in this study. This study shows the results that have been collected through the study of the following methods. Out of 83 samples, only 25 sponges were successfully identified. They were from class Demospongia and were under subclass Heteroscleremorpha. The majority of the two main orders are Haplosclerida (28%) and order Tetractinellida (24%). The rest were from order Clionaida and Scopalinida (12%), Suberitida (8%), while order Axinellida, Tethyida, Poecilosclerida and Agelasida (4%). Among the identified samples, 16 were identified to species level while the rest were only identified up to genus level. A significant limitation of this study is that the sequences in the database are not well populated and have low diversity, making it challenging to conduct molecular identification. It also impacted the accuracy of the classification, making a single method of identification less reliable.

Distribution of marine sponges based on depth area of Mesophotic Coral Ecosystems (MCEs) in North Lombok Waters, NTB Province

North Lombok Waters have a fairly high level of diversity. Mesophotic Coral Ecosystems (MCEs) are light-dependent coral reef communities ranging from 30-40 meters and extending to the bottom of the photic zone to a depth of 150 meters. Sponges are the main constituent of MCEs that have many benefits, but their existence is poorly understood. This study aims to determine the distribution of marine sponges in Mesophotic Coral Ecosystems (MCEs). Data was taken in September 2020 using the Underwater Photo Transect (UPT) method. Pictures were taken at a depth of 10, 20, 30, 40, and 50 meters at 3 stations. Macroscopic identification of sponges based on underwater photos. The identification results found 32 genera of sponges spread over 3 stations in the waters of North Lombok. The deeper you go, the more genera are found at the three research stations. Out of 32 genera, sponge abundance dependent on depth only influences 5 genera. We anticipate additional factors affecting the 27 genera of sponges, but further research is necessary. The ANOVA test results showed that several sponges genera were significantly different between depths, namely Callyspongia, Theonella, Clathria, Stylotella, and Spirastrella.

Taxonomical identification of marine sponges from crab net bycatch

Taxonomical identification of marine sponges from crab net bycatch

Keratose sponges in ancient carbonates – A problem of interpretation

AbstractReports of diverse vermiform and peloidal structures in Neoproterozoic to Mesozoic open marine to peritidal carbonates include cases interpreted to be keratose sponges. However, living keratose sponges have elaborate, highly elastic skeletons of spongin (a mesoscopic end‐member of a hierarchical assemblage of collagenous structures) lacking spicules, thus have poor preservation potential in contrast to the more easily fossilized spicule‐bearing sponges. Such interpreted fossil keratose sponges comprise diverse layered, network, amalgamated, granular and variegated microfabrics of narrow curved, branching, vesicular–cellular to irregular areas of calcite cement, thought to represent former spongin, embedded in microcrystalline to peloidal carbonate. Interpreted keratose sponges are presented in publications almost entirely in two‐dimensional (thin section) studies, usually displayed normal to bedding, lacking mesoscopic three‐dimensional views in support of a sponge body fossil. For these structures to be keratose sponges critically requires conversion of the spongin skeleton into the calcite cement component, under shallow‐burial conditions and this must have occurred prior to compaction. However, there is no robust petrographic–geochemical evidence that the fine‐grained carbonate component originated from sponge mummification (automicritic body fossils via calcification of structural tissue components) because in the majority of cases the fine‐grained component is homogenous and thus likely to be deposited sediment. Thus, despite numerous studies, verification of fossil keratose sponges is lacking. Although some may be sponges, all can be otherwise explained. Alternatives include: (i) meiofaunal activity; (ii) layered microbial (spongiostromate) accretion; (iii) sedimentary peloidal to clotted micrites; (iv) fluid escape and capture resulting in bird's eye to vuggy porosities; and (v) moulds of siliceous sponge spicules. Uncertainty of keratose sponge identification is fundamental and far‐reaching for understanding: (i) microfacies and diagenesis where they occur; (ii) fossil assemblages; and (iii) wider aspects of origins of animal clades, sponge ecology, evolution and the systemic recovery from mass extinctions. Thus, alternative explanations must be considered.

The Utilityof 28S rDNA for Barcoding of Freshwater Sponges (Porifera, Spongillida)

Sponges (Porifera, Spongillida) make up the bulk of the benthic biomass in Lake Baikal and are represented by the family Lubomirskiidae, a collection of endemic species, and several species of the cosmopolitan family Spongillidae. We conducted an analysis of the D3 domain of the 28S rDNA of 16 freshwater sponge species. Based on molecular data, we were able to identify all of the collected Spongillidae specimens whose identification was difficult due to the lack of gemmules. Phylogenetic trees have shown that Ephydatia muelleri, Spongilla lacustris, and Eunapius fragilis formed monophyletic clades, and the D3 domain of the 28S rDNA can be used for their DNA barcoding. For the Baikal sponges, the use of this marker is important since the gemmule-less Spongillidae and Lubomirskiidae are, in some cases, indistinguishable from each other in morphology. The 28S rDNA has been shown to be useful for family and species-level identification of freshwater sponges within the Spongillida.

Screening and molecular identification of marine sponges with cytotoxic activities, collected from Gulf of Mannar, Indian Ocean

The marine environment, a rich source of biological and chemical diversity, offers tremendous scope for discovering novel metabolites with pharmacological activities. Marine invertebrates, especially sponges, have gained more attention in drug discovery programmes since they exhibit unique metabolic and physiological capabilities and an extraordinarily symbiotic association with diverse bacterial communities. The present study focuses on screening the cytotoxic activity of the methanolic extracts of sponges against the Non-Small Cell Lung Cancer (NSCLC) cell line, NCI-H460. In the bioactivity screening, two sponges (IOS-11 and IOS-12) were found with potent cytotoxic activity. The sponge IOS-12 showed almost complete cell growth inhibition (99 %) at 50 µg/ml and 80 % inhibition at 5 µg/ml; whereas IOS-11 showed cell growth inhibition of 96 % and 77 % at 50 µg/ml and at 5 µg/ml, respectively. Amplification of the mitochondrial cytochrome C oxidase subunit I (COI) and subsequent nucleotide analysis enabled the identification of sponge samples, IOS-11 and IOS-12, as Rhabdastrella globostellata and Halichondria sp., respectively. The study concludes that the two marine sponges, Rhabdastrella globostellata and Halichondria sp., possess excellent cytotoxic activities and could be considered good candidates for the isolation of anticancer compounds with biomedical applications. The isolation, purification and structural elucidation of the lead molecules from these sponges are ongoing studies, which might yield potential new compounds with promising cytotoxic activities.

DNA Barcoding Procedures for Taxonomical and Phylogenetic Studies in Marine Animals: Porifera as a Case Study.

DNA barcoding is a versatile approach that has revolutionized taxonomy and other akin topics in biology and ecology, due to its simplicity and relatively costless procedures. The method consists in the production of one or a few amplicons from informative genetic regions via Sanger sequencing. These markers are selected because they tend to evolve at a similar pace as speciation, allowing to discriminate organismal species. The applicability of this technique is here portrayed for the taxonomical identification of marine sponges (phylum: Porifera) as an exemplification.

Hundreds of new DNA barcodes for South African sponges

DNA barcoding based on a fragment of the cytochrome c oxidase subunit I (COI) gene from the mitochondrial genome is widely applied in species identification, species discovery and biodiversity studies. The aim of this study was to establish a barcoding reference database of sponges collected from South Africa, and evaluate the applicability of the COI gene for aiding in the identification of sponges in combination with tentative morphological identifications. A total of 317 mitochondrial COI barcode sequences, with an additional 21 extended COI fragments and 24 nuclear ITS sequences, were obtained from 11 orders, 38 families, 58 genera and 124 species of spiculated sponges. A Neighbour Joining (NJ) trees that were reconstructed using these sequences in most cases clustered species in accordance with their current taxonomic identification, and we conclude that COI sequencing can be used to aid in the identification of sponge species. We further demonstrate that DNA barcoding analysis has potential to uncover cryptic sponge species, and to reveal dubious morphological identifications. We recommend that future taxonomic studies of South African sponges incorporate multiple sources of information for species identification or discovery.

Immunomodulatory Activity of the Marine Sponge, Haliclona (Soestella) sp. (Haplosclerida: Chalinidae), from Sri Lanka in Wistar Albino Rats: Immunosuppression and Th1-Skewed Cytokine Response.

Natural secondary metabolites of sponges of the genus Haliclona are associated with an array of biological activity with therapeutic usage. We investigated the immunopharmacological properties of a presumably novel marine sponge species from Sri Lanka, Haliclona (Soestella) sp. Sponge material was collected from southern Sri Lanka by scuba diving. Sponge identification was based on spicule and skeleton morphology using light microscopy. Selected in vivo and ex vivo tests investigated nonfunctional and functional immunomodulatory activity of the Haliclona (Soestella) sp. crude extract (HSCE) in the Wistar rat model. Compared to the controls, rats orally gavaged daily for 14 consecutive days with 15 mg/kg dose of the HSCE manifested a significant reduction of immune cell counts of total WBCs (by 17%; p < 0.01), lymphocytes (38%), platelets (52%), splenocytes (20%), and bone marrow cells (BMC; 60%) (p < 0.001), with a concurrent increase in the neutrophil : lymphocyte ratio (p < 0.05); RBC counts abated by 53% (p < 0.001). A significant reduction of the splenosomatic index was evident with the 10 and 15 mg/kg doses (p < 0.001). Rat plasma TNF-α cytokine level was augmented by tenfold (p < 0.001), IL-6 level by twofold (p < 0.01) with the 15 mg/kg HSCE treatment, while IL-10 was detectable in rat plasma only with this treatment; the corresponding Th1 : Th2 cytokine ratio (TNF-α : IL-10) was indicative of an unequivocal Th1-skewed cytokine response (p < 0.01). Ex vivo bone marrow cell and splenocyte proliferation were significantly and dose dependently impaired by HSCE (IC50 0.719 and 0.931 μg/mL, respectively; p < 0.05). Subacute toxicity testing established that HSCE was devoid of general toxic, hepatotoxic, and nephrotoxic effects. In conclusion, HSCE was orally active, nontoxic, and effectively suppressed nonfunctional and functional immunological parameters of Wistar rats, suggestive of the potential use of the HSCE as an immunosuppressant drug lead.

Calcareous sponges from the French Polynesia (Porifera: Calcarea).

Although the French Polynesian reefs are among the most well studied reefs of the world, sponges are still poorly known, with only 199 species or OTUs of sponges having been described from French Polynesia, 167 at an OTU level and 32 at a species level. From those 199 species, just five are calcareous sponges. As it is possible that this number is underestimated, the aim of the present work was to study the diversity of calcareous sponges from French Polynesia. Hence, different French Polynesian archipelagos were surveyed by SCUBA from 3 to 60 m of depth. Identifications were performed using morphological and molecular (ITS and C-LSU) tools. We found a total of nine species of Calcarea, comprising five different genera. Five species are new to science: Clathrina fakaravae sp. nov., Clathrina huahineae sp. nov., Ernstia variabilis sp. nov., Leucascus digitiformis sp. nov., and Leucandra tahuatae sp. nov. With the present work, the number of identified sponges from French Polynesia at a species level increased from 32 to 41. The only calcareous sponge previously known from French Polynesia that was recollected by our group was Leucetta chagosensis. Our results suggest that the Eastern Indo-Pacific Realm shows more affinity with the Central and the Western Indo-Pacific Realms. Four species supported these affinities: Ascandra cf. crewsi, previously known only from Papua New Guinea, Leucascus simplex from South Australia, and Leucetta chagosensis and L. microraphis, both widespread species in the Indo-Pacific. These two Leucetta species, however, most likely represent species complexes. Once again the molecular markers ITS and C-LSU helped in the identification of calcareous sponges, showing how important is an integrative taxonomy. Although our work has increased in 250% (6 spp to 15 spp) the diversity of calcareous sponges in French Polynesia, it is most possible that this number is still underestimated.

Intragenomic variation of rDNA internal transcribed spacers in the endemic Baikal sponge Lubomirskia baikalensis (Pallas, 1776) (Spongillida, Lubomirskiidae): Implications for Porifera barcoding

Baikal endemic sponges are a species group for which species identification is difficult and the taxonomy is insufficiently developed. The rDNA ITS (internal transcribed spacers) are practical molecular markers for sponges. However, intragenomic polymorphism of the ITS spacers has recently been detected in many organisms including marine and freshwater sponges. We for the first time demonstrate intragenomic polymorphism of the ITS region in the sponge family Lubomirskiidae. We identified 27 new ITS1 and ITS2 haplotypes. The level of intragenomic polymorphism of the ITS1 and ITS2 regions in Lubomirskia baikalensis was 0.1–2.0%, which overlaps with the intraspecific polymorphism of L. baikalensis and to the interspecific polymorphism within Lubomirskiidae. Thus, the detected level of intragenomic polymorphism of Lubomirskiidae suggests species identification of Baikal sponges using ITS sequences would be difficult. The reasons for the lack of concerted evolution of the ITS spacers in Lubomirskiidae may be hybridization or insufficient divergence time. Our work contributes to the development of sponge barcoding markers.

UJI AKTIVITAS ANTIBAKTERI BEBERAPA SPONS DARI PERAIRAN SALIBABU KEPULAUAN TALAUD

Sponge is one component of coral reefs that has the potential as a bioactive compound but has not been widely used. This study was directed to obtain some spongy crude extracts from the Salibabu waters of the Talaud Islands, determine the antibacterial activity of some crude extruded sponges and compare the antibacterial activity of the sponge fraction against E. coli and S. aureus test bacteria. The results of identification of sponges were found in three species consisting of: Siphonodictyon sp., Ircinia sp., Dysidea sp. The antibacterial activity of crude extract, water, methanol and hexane fractions using agar diffusion method with E. coli and S. aureus test bacteria. The crude extract of siphonodictyon sp and the water fraction of this extract show the highest antibacterial activity. &#x0D; Key words: Spons, Antibacterial, E. coli. S. aureus

Minimalist barcodes for sponges: a case study classifying African freshwater Spongillida.

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.

Molecular taxonomy of some sponges (Demospongiae) using ribosomal (18s rrna) and partial mitochondrial (coi) genes

Sponges, the most ancient multicellular metazoan, were widely distributed across habitats. Vietnam is known to possess a high biodiversity of sponges, however, they are mostly identified based on morphological characteristics and lack the molecular data. In the current study, the phylogenetic relationship of some sponges (Demospongiae) in Vietnam was constructed using two independent markers (COI and 18S rRNA). In this paper the individual markers (COI and 18S rRNA) were successfully used to identify some sponge taxa at the species level. The obtained results showed the congruence of molecular taxonomy using two independent markers. However, our study showed that a combination of the two markers provided more information and supported better for sponge identification. At order level, the COI tree and 18S rRNA tree also recovered the same clades, indicating the congruence of COI and 18S rRNA genes in sponge classification. However, branching order of the clades in COI tree was weakly supported and slightly different from those in 18S rRNA tree.

Unravelling the moons: review of the genera Paratetilla and Cinachyrella in the Indo-Pacific (Demospongiae, Tetractinellida, Tetillidae).

Paratetillabacca (Selenka, 1867) and Cinachyrellaaustraliensis (Carter, 1886) occur in a broad range of marine environments and are allegedly widely distributed species in the Indo-Pacific. We coin the term ‘moon sponges’ for these species as they are spherical in shape with numerous porocalices resembling the lunar surface. Both species have a complex taxonomic history with high synonymization, in particular by Burton (1934, 1959). An examination of the junior synonyms proposed by Burton (1934, 1959) was conducted to establish the validity of the names. More than 230 specimens from Naturalis Biodiversity Center were reviewed that belong to the genera Paratetilla and Cinachyrella from marine lakes, coral reefs, and mangroves in Indonesia. The aim of the current study was to untangle the taxonomic history, describe the collection of moon sponges from Indonesia, and develop a key. We extensively reviewed the taxonomic literature as well as holotypes of most of the species synonymized by Burton. The taxonomic history of Paratetilla spp. and Cinachyrellaaustraliensis showed some cases of misinterpreted synonyms, misidentifications, and lack of detailed descriptions for some species. The conclusion of the revision is that there are three valid species of Paratetilla (P.arcifera, P.bacca, and P.corrugata) and four valid species of Cinachyrella (C.australiensis, C.porosa, C.paterifera, and C.schulzei) in Indonesia. This is furthermore corroborated by molecular work from previous studies. ParatetillaarciferaWilson 1925 and C.porosa (Lendenfeld, 1888) are resurrected. A full review of taxonomic history is provided as well as a key for identification of moon sponges from Indonesia. All species are sympatric and we expect that there are undescribed species remaining within the Tetillidae from the Indo-Pacific. Our current review provides the framework from which to describe new species in the genera Paratetilla and Cinachyrella from the Indo-Pacific.

Unravelling the moons: review of the genera Paratetilla and Cinachyrella in the Indo-Pacific (Demospongiae, Tetractinellida, Tetillidae)

Paratetillabacca(Selenka, 1867) andCinachyrellaaustraliensis(Carter, 1886) occur in a broad range of marine environments and are allegedly widely distributed species in the Indo-Pacific. We coin the term ‘moon sponges’ for these species as they are spherical in shape with numerous porocalices resembling the lunar surface. Both species have a complex taxonomic history with high synonymization, in particular by Burton (1934, 1959). An examination of the junior synonyms proposed by Burton (1934, 1959) was conducted to establish the validity of the names. More than 230 specimens from Naturalis Biodiversity Center were reviewed that belong to the generaParatetillaandCinachyrellafrom marine lakes, coral reefs, and mangroves in Indonesia. The aim of the current study was to untangle the taxonomic history, describe the collection of moon sponges from Indonesia, and develop a key. We extensively reviewed the taxonomic literature as well as holotypes of most of the species synonymized by Burton. The taxonomic history ofParatetillaspp. andCinachyrellaaustraliensisshowed some cases of misinterpreted synonyms, misidentifications, and lack of detailed descriptions for some species. The conclusion of the revision is that there are three valid species ofParatetilla(P.arcifera,P.bacca, andP.corrugata) and four valid species ofCinachyrella(C.australiensis,C.porosa,C.paterifera, andC.schulzei) in Indonesia. This is furthermore corroborated by molecular work from previous studies.ParatetillaarciferaWilson 1925 andC.porosa(Lendenfeld, 1888) are resurrected. A full review of taxonomic history is provided as well as a key for identification of moon sponges from Indonesia. All species are sympatric and we expect that there are undescribed species remaining within the Tetillidae from the Indo-Pacific. Our current review provides the framework from which to describe new species in the generaParatetillaandCinachyrellafrom the Indo-Pacific.

Identification of an aquaculture poriferan "Pest with Potential" and its phylogenetic implications.

Correct identification and classification of sponges is challenging due to ambiguous or misleading morphological features. A particular case is a blue keratose sponge occasionally referred to as the “Blue Photo Sponge” among aquarists, which appears frequently (and in several cases unintended) in private aquaria. This spicule-less species, occasionally specified as Collospongia auris Bergquist, Cambie & Kernan 1990, not only displays a high phenotypic plasticity in growth form and colour, it also proliferates in aquacultures under standard conditions unlike most other sponges. Therefore, this species is regarded as a pest for most aquarists. In turn, the ease of cultivation and propagation in aquacultures qualifies this species as a model organism for a wide array of scientific applications. For these purposes, correct identification and classification are indispensable. We reconstructed ribosomal gene trees and determined this species as Lendenfeldia chondrodes (De Laubenfels, 1954) (Phyllospongiinae), distant to Collospongia auris, and corroborated by skeletal features. Additionally, the resulting phylogeny corroborated major shortcomings of the current Phyllospongiinae classification—its consequences are discussed.

Contribution to the knowledge on the distribution of freshwater sponges – the Danube and Sava rivers case study

Sponges in the large rivers within the Danube River Basin (DRB) have not been adequately studied. Hence, the aim of this work was to undertake an investigation on the distribution of sponge species in the Danube and Sava rivers. Out of 88 localities covered by the study, sponges were found at 25 sites only (46 samples in total). By using morphological (light and scanning electron microscopy) and genetic (28S rDNA sequencing) analysis, four species were determined: Ephydatia fluviatilis (Linnaeus, 1759), Spongilla lacustris (Linnaeus, 1759), Eunapius fragilis (Leidy, 1851), Trochospongilla horrida Weltner, 1893. In the Danube, the predominant species was found to be E. fluviatilis making approximately 80% of collected samples, while in the Sava River S. lacustris dominated, representing 46% of the river sponges. Our work represents one of the few studies on freshwater sponges within the DRB from long stretches of the large lowland rivers (more than 2500 km of the Danube River and about 900 km of the Sava River). Moreover, molecular analysis for the identification of freshwater sponges was applied on the material collected from a wide area, thus contributing to the systematic studies on the distribution and abundance of the European freshwater invertebrate fauna in general.

Transformation of Baikal sponges (family Lubomirskiidae) after penetration into the Angara River

Genetic and morphological analysis of endemic sponges of the Lubomirskiidae family from Lake Baikal and the upper reaches of the Angara River was performed. Various sponge species acquired a number of similar morphological traits after the transition from the lake into the river. These traits enabled an increase of sponge skeleton strength under the conditions of elevated hydrodynamic activity. The changes significantly impeded morphology-based species identification of Angara sponges. Phylogenetic analysis of ITS regions and noncoding mitochondrial DNA fragments confirmed that the Angara sponges belonged to the Baikalian Lubomirskiidae family and demonstrated the polyphyletic origin of the sponges. The use of combined molecular and morphological data allowed for the clustering of some sponge samples into groups that corresponded to individual species. The absence of genetic isolation between the Baikalospongia intermedia and Lubomirskia baicalensis species was demonstrated, whereas the B. intermedia profundalis subspecies was well separated from B. intermedia. This finding pointed to the necessity of further studies for the clarification of the taxonomic status of this subspecies.

Development of a multilocus-based approach for sponge (phylum Porifera) identification: refinement and limitations

For sponges (phylum Porifera), there is no reliable molecular protocol available for species identification. To address this gap, we developed a multilocus-based Sponge Identification Protocol (SIP) validated by a sample of 37 sponge species belonging to 10 orders from South Australia. The universal barcode COI mtDNA, 28S rRNA gene (D3–D5), and the nuclear ITS1-5.8S-ITS2 region were evaluated for their suitability and capacity for sponge identification. The highest Bit Score was applied to infer the identity. The reliability of SIP was validated by phylogenetic analysis. The 28S rRNA gene and COI mtDNA performed better than the ITS region in classifying sponges at various taxonomic levels. A major limitation is that the databases are not well populated and possess low diversity, making it difficult to conduct the molecular identification protocol. The identification is also impacted by the accuracy of the morphological classification of the sponges whose sequences have been submitted to the database. Re-examination of the morphological identification further demonstrated and improved the reliability of sponge identification by SIP. Integrated with morphological identification, the multilocus-based SIP offers an improved protocol for more reliable and effective sponge identification, by coupling the accuracy of different DNA markers.