Investigation of the budding process in Tethya citrina and Tethya aurantium (Porifera, Demospongiae)

The budding process has been studied in two congeneric Mediterranean species belonging to Tethya from different sampling sites: Marsala and Venice Lagoons (Tethya citrina); Marsala Lagoon and Porto Cesareo Basin (Tethya aurantium). Buds, connected to the adult by a spiculated stalk, differ between the two species in morphology and size, since those of T. citrina are small with elongated bodies, showing only a few spicules protruding from the apical region, whereas those of T. aurantium are round, larger, and show spicules radiating from the peripheral border. In T. citrina, cells with inclusions, varying in electron density and size, represent the main cell types of the buds. In T. aurantium, the cell component shows a major diversification, resulting from spherulous cells, grey cells, vacuolar cells and peculiar micro-vesicle cells. Neither canals nor choanocyte chambers were observed in the buds of the two species. In T. citrina, bud production is similar in both sampling sites. In T. aurantium, budding occurs more rarely in Porto Cesareo Basin, probably in relation with environmental factors, such as the covering of the cortex by sediment and micro-algae. Finally, in the buds of both species, the spicule size does not differ from that of the cortex of the adult sponges, further supporting the main involvement of the cortex in organizing the skeletal architecture of the buds.     

Redescription of Tethya norvegica Bowerbank (Porifera, Demospongiae), with remarks on the genus Tethya in the North East Atlantic

Tethya norvegica Bowerbank, 1872, a northern species generally synonymized with T. aurantium . has been studied from Scotland, The Faroes, Norway, Iceland, Spitzbergen and Bear Island. The species is redescribed and its North East Atlantic distribution delimited Tethya norvegica is compared, on the basis of morphometric and electrophoretic features, with T. aurantium and F.citrina . Morphological, morphometric and electrophoretic data suggest that I norvegica is a distinct biological species.     

Association of the sponge Tethya orphei (Porifera, Demospongiae) with filamentous cyanobacteria

Abstract. Specimens of the sponge Tethya orphei , collected in February 2005 on the underside of coral stones on Arì Athol (Maldives), have been processed for histological and ultrastructural investigations. The cortical layer of the sponge was found to be permeated by filamentous cyanobacteria, the trichomes of which measured 45–63 μm on average and were composed of 10–14 cells. The fine organization of the filaments was consistent with their taxonomic identification as Oscillatoria spongeliae. These filaments filled the cortical region of the sponge and penetrated inward into the upper choanosomal region, where they sometimes overlapped the siliceous spicule bundles. A budding specimen of T. orphei showed that the filaments were also present in the single bud protruding from the sponge surface, demonstrating that asexual reproduction can vertically transmit these symbionts from sponge to sponge. The occurrence of filaments in all the specimens studied is consistent with the assumption that filamentous cyanobacteria are not mere intruders but mutualistic symbionts with members of T. orphei.     

Revision of the genus Clathrina (Porifera, Calcarea)

The genus Clathrina has one of the most difficult systematic arrangements in the Porifera, Class Calcarea. Few morphological characters can be used to describe its species, and the systematics and the geographical distribution of its species have changed several times, according to the point of view of the systematists. 'Lumpers' consider that clathrinas are morphologically plastic, while 'splitters' believe that even slight morphological differences should be considered sufficient to distinguish between species. The morphology of several specimens/species of Clathrina , including the type species, when possible, was studied and used to produce the first revision of this genus. Using results obtained from previous morphological and molecular studies, the morphological characters were chosen and analysed in all studied specimens. In total, 43 species were found, nine of which are new to science. These results agree with the viewpoint that morphological characters such as type and distribution of spicules, size of actines, spines, anastomosis of the cormus, organization of the osculum and presence of granules in cells, are useful when establishing the taxonomy of the genus.  © 2003 The Linnean Society of London, Zoological Journal of the Linnean Society , 2003, 139 , 1−62.     

Interspecific variation in arrangement and morphology of micrasters of Tethya species (Porifera,Demospongiae)

Summary A new distinctive feature between the two Mediterranean species of Tethya, T. aurantium and T. citrina has been found in the body arrangement of different types of micrasters. Contrary to the previous assumptions, T. aurantium has two clearly distinct categories of micrasters: the chiaster-tylaster in the cortex and the larger, slender oxyaster in the choanosome. T. citrina has only slightly differentiated micraster sets in the cortex and choanosome; in the latter the shape of micrasters is close to that of oxyasters. SEM analysis shows that differences in micraster shape depend on the cylindrical or conical form of rays and on the distribution, density and strength of the microspines along their axis. The relationship between the degree of micraster differentiation and the development of the cortex in the two species is discussed.     

Allograft rejection, autograft fusion and inflammatory responses to injury in Callyspongia diffusa (Porifera; Demospongia)

Sponges exhibit a variety of swift, cellular defence responses to protect self integrity. The sponge Callyspongia diffusa has been used to characterize the cytological changes that occur during allograft rejection, autograft fusion, and inflammation. Allogeneic contact results in fusion of the two exopinacoderms followed by an infiltration of mesohyl cells into the graft zone. As mesohyl cells accumulate, they form tissue bridges that span the graft interface. After a few days, the tissue bridges and the nearby cellular infiltrate become necrotic and slough off, which separates the allogeneic tissues. Autograft fusion begins similarly but cellular infiltration does not follow exopinacoderm fusion. Contacted exopinacocytes are redistributed, the endopinacoderms and choanosomes come into contact, and the grafted sponge tissues merge. Tissue damage exposes internal regions of the sponge to the external environment. In many areas of injury, exposed choanosome is sealed by infiltrating mesohyl cells. In other areas, exposed endopinacoderm appears to serve as new exopinacoderm. Cellular debris is removed by phagocytic archaeocytes and new exopinacoderm is regenerated over the damaged choanosome. No scars remain once the inflammatory infiltrate has dispersed. In general, mesohyl cells are involved in defence responses without an observed enrichment of any specific cell type. However, archaeocytes from rejecting sponges appear to line both sides of the allogeneic interface.     

Functional morphology of Tethya species (Porifera): 1. Quantitative 3D-analysis of Tethya wilhelma by synchrotron radiation based X-ray microtomography

Rhythmic body contraction is a phenomenon in the Porifera, which is only partly understood. As a foundation for the understanding of the functional morphology of the highly contractile Tethya wilhelma, we performed a qualitative and quantitative volumetric 3D-analysis of the morphology of a complete non-contracted specimen at resolutions of 5.2 and 6.9 μm, using synchrotron radiation based X-ray computed microtomography (SR-μCT). For the first time, we were able to visualize all three major body structures of a complete poriferan without dissection of the shock-frozen, fixed and contrasted specimen in a near-to-life confirmation: poriferan tissue, mineral skeleton and aquiferous system. Applying a ‘virtual cast’ technique allowed us to analyze the structural details of the complete canal structure. Our results imply an extensive re-circulation of water inside the poriferan due to well-developed by-pass-canals, connecting excurrent and incurrent system. Nevertheless, the oscule region is strictly separated from the incurrent system. Based on our data, we developed a hypothetical flow regime for T. wilhelma, which explains the necessity of by-pass canals to minimize pressure boosts in the canal system during contraction. Additionally, re-circulation optimizes nutrient uptake, within small-sized poriferans, like T. wilhelma. Quantitative analysis allowed us to measure volumes and surfaces, displaying remarkable organizational differences between choanosome and cortex, by means of distribution of morphological elements. The surface-to-volume ratio proved to be very high, underlining the importance of the poriferan pinacoderm. We support a pinacoderm-contraction hypothesis.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .Dedicated to Prof. Dr. Michele Sarà (Genova, Italy), in honour of his 80th birthday in 2006.     

Neuroactive substances specifically modulate rhythmic body contractions in the nerveless metazoon Tethya wilhelma (Demospongiae, Porifera)

Background

Mitochondrial (mt) gene arrangement is highly variable among molluscs and especially among bivalves. Of the 30 complete molluscan mt-genomes published to date, only one is of a heterodont bivalve, although this is the most diverse taxon in terms of species numbers. We determined the complete sequence of the mitochondrial genomes of Acanthocardia tuberculata and Hiatella arctica, (Mollusca, Bivalvia, Heterodonta) and describe their gene contents and genome organisations to assess the variability of these features among the Bivalvia and their value for phylogenetic inference.

Results

The size of the mt-genome in Acanthocardia tuberculata is 16.104 basepairs (bp), and in Hiatella arctica 18.244 bp. The Acanthocardia mt-genome contains 12 of the typical protein coding genes, lacking the Atpase subunit 8 (atp8) gene, as all published marine bivalves. In contrast, a complete atp8 gene is present in Hiatella arctica. In addition, we found a putative truncated atp8 gene when re-annotating the mt-genome of Venerupis philippinarum. Both mt-genomes reported here encode all genes on the same strand and have an additional trnM. In Acanthocardia several large non-coding regions are present. One of these contains 3.5 nearly identical copies of a 167 bp motive. In Hiatella, the 3' end of the NADH dehydrogenase subunit (nad)6 gene is duplicated together with the adjacent non-coding region. The gene arrangement of Hiatella is markedly different from all other known molluscan mt-genomes, that of Acanthocardia shows few identities with the Venerupis philippinarum. Phylogenetic analyses on amino acid and nucleotide levels robustly support the Heterodonta and the sister group relationship of Acanthocardia and Venerupis. Monophyletic Bivalvia are resolved only by a Bayesian inference of the nucleotide data set. In all other analyses the two unionid species, being to only ones with genes located on both strands, do not group with the remaining bivalves.

Conclusion

The two mt-genomes reported here add to and underline the high variability of gene order and presence of duplications in bivalve and molluscan taxa. Some genomic traits like the loss of the atp8 gene or the encoding of all genes on the same strand are homoplastic among the Bivalvia. These characters, gene order, and the nucleotide sequence data show considerable potential of resolving phylogenetic patterns at lower taxonomic levels.     

Differences between two sympatric species of Tethya (Porifera, Demospongiae) concerning the growth and final form of their megasters

The megasters of T. aurantium and T. citrina consist, in their mature stage, of spherasters and spheroxyasters respectively. A study of the ratio between the ray length and the diameter of the centre (R/C), which determines the megaster shape, has been carried out on spicules of different size.
While the small megasters of these two species have a similar shape, successive stages show in T. aurantium an isometric growth and in T. citrina an allometric growth with rays growing faster than the centre. The two species differ also in the mean size, size frequency distribution and number of rays of the asters. Differences in the mean total diameter and shape were also detected among spicules coming from different regions of the sponge body. The taxonomic and evolutionary meaning of these phenomena are discussed.     

Allogeneic cell interactions during graft rejection in Callyspongia diffusa (Porifera, Demospongia); a study with monoclonal antibodies

Many aspects of the cellular immune system in the marine sponge Callyspongia diffusa, have been defined by using artificially transplanted allogeneic tissues. Rejections show specificity of 'non-self' recognition, cytotoxic effector responses and short-term immunological memory. Histological investigations reveal a generalized mesohyl migration to the graft zone where archaeocytes line up at the allogeneic interface. Monoclonal antibodies (mAbs) raised to sponge cells have shown that little or no allogeneic cell mixing occurs at the graft interface and that certain mesohyl cell types do not appear to be directly involved in graft rejections. However, all mesohyl cell types are present in autograft fusion zones and in inflammatory responses to injury. The involvement of only some of the mesohyl cell types in graft rejections suggests specific interactions of an effector 'immunocyte'.     

Observations on the genus Indiana Chakravarty, 1943 (Oxyurida: Pulchrocephalidae)

Summary Indiana gryllotalpae is redescribed from the gut of a mole-cricket, Scapteriscus sp., from Trinidad. The structure of the cephalic umbraculum (comprising 6 major and 6 minor cuticular elements extending back from the setulose lip region) is described in detail by use of both light and scanning electron microscopy. I. gryllotalpae of Bain (1965) is regarded as a synonym of I. coimbatoriensis Latheef & Seshadri, 1972. ac]19820729     

Functional morphology of Tethya species (Porifera): 2. Three-dimensional morphometrics on spicules and skeleton superstructures of T. minuta

The biomechanics of body contraction in Porifera is almost unknown, although sponge contraction has been observed already in ancient times. Some members of the genus Tethya represent the most contractile poriferan species. All of them show a highly ordered skeleton layout. Based on three main spicule types, functional units are assembled, termed skeleton superstructures here. Using synchrotron radiation based x-ray microtomography and quantitative image analysis with specially developed particle and structure recognition algorithms allowed us to perform spatial allocation and 3D-morphometric characterizations of single spicules and skeleton superstructures in T. minuta. We found and analyzed three skeleton superstructures in the investigated specimen: (1) 85 megasclere bundles, (2) a megaster sphere, composed by 16,646 oxyasters and (3) a pinacoderm–tylaster layer composed by micrasters. All three skeleton superstructures represent composite materials of siliceous spicules and extracellular matrix. From structure recognition we developed an abstracted mathematical model of the bundles and the sphere. In addition, we analyzed the megaster network interrelation topology and found a baso-apical linear symmetry axis for the megaster density inside the sphere. Based on our results, we propose a hypothetical biomechanical contraction model for T. minuta and T. wilhelma, in which the skeleton superstructures restrain physical stress generated by contraction in the tissue. While skeletal structures within the genus Tethya have been explained using R. Buckminster Fullers principle of tensegrity by other authors, we prefer material science based biomechanical approaches, to understand skeletal superstructures by referring to their composite material properties.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .     

A new genus and species of deep-sea glass sponge (Porifera, Hexactinellida, Aulocalycidae) from the Indian Ocean

New hexactinellid sponges were collected from 2589 m depth on the Carlsberg Ridge in the Indian Ocean during deep-sea dredging. All fragments belong to a new genus and species, Indiellagen. n.ridgenensissp. n., a representative of the family Aulocalycidae described here. The peculiar features of this sponge, not described earlier for other Aulocalycidae, are: longitudinal strands present in several layers and epirhyses channelization.     

Cytochemical Observations of Gemmule Development in Eunapius fragilis (Leidy): Porifera; Spongillidae

The cytology and cytochemistry of gemule formation in the freshwater sponge Eunapius fragilis (Leidy) is investigated. The aggregation of ameboid cells which initiates gemmulation is composed of three distinct cell types, two of these fitting the morphological criteria of the archeocyte class. The third cell type, the granular cell, is a differentiated cell type which undergoes cytolysis during gem-mule coat synthesis. Following early polarisation of the aggregate into an internal reproductive area and a peripheral coat region, a structurally and chemically heterogeneous gemmule coat is synthesis-ed. The coat is formed through secretory activities involving several cell types including, possibly, the internal endothelium. The coat is composed of "collagenous" fibers embedded in a matrix of varying degrees of density–showing close structural and cytochemical similarities to vertebrate chondroid tissue. Vitelline platelets of thesocytes of E. fragilis do not cytochemically resemble any previously reported vitelline inclusions from other sponges. Outer valves of the platelets contain components stained by basic dyes at acidic pH which are not extracted by RNase. Distinct differences in developmental processes and morphological–including cytochemical–characteristics of gemmules of different spongillid species are apparent.     

Larval development in the Homoscleromorpha (Porifera, Demospongiae)

Abstract. Embryonic development from coeloblastula to fully developed larva was investigated in 8 Mediterranean homoscleromorph species: Oscarella lobularis, O. tuberculata, O. microlobata, O. imperialis, Plakina trilopha, P. jani, Corticium candelabrum , and Pseudocorticium jarrei. Morphogenesis of the larva is similar in all these species; however, cell proliferation is more active in species of Oscarella than in Plakina and C. candelabrum. The result of cell division is a wrinkled, flagellated larva, called a cinctoblastula. It is composed of a columnar epithelium of polarized, monoflagellated cells among which are scattered a few non-flagellated ovoid cells. The central cavity always contains symbiotic bacteria. Maternal cells are also present in O. lobularis, O. imperialis , and P. jarrei. In the fully developed larva, cell shape and dimensions are constant for each species. The cells of the anterior pole have large vacuoles with heterogeneous material; those of the postero-lateral zone have an intranuclear paracrystalline inclusion; and the flagellated cells of the posterior pole have large osmiophilic inclusions. Intercellular junctions join the apical parts of the cells, beneath which are other specialized cell junctions. A basement membrane underlying the flagellated cells lines the larval cavity. This is the first observation of a basement membrane in a poriferan larva. The basal apparatus of flagellated cells is characterized by an accessory centriole located exactly beneath the basal body. The single basal rootlet is cross striated. The presence of a basement membrane and a true epithelium in the larva of Homoscleromorpha—unique among poriferan clades and shared with Eumetazoa—suggests that Demospongiae could be paraphyletic.     

Tissue organization ofFarrea occa (Porifera,Hexactinellida)

Summary The tissue organization ofFarrea occa has been examind by light microscopy, transmission electron microscopy (TEM), and scanning electron microscopy (SEM). It was found to agree closely with the hexactinellid model established forRhabdocalyptus dawsoni by Mackie and Singla (1983) in consisting of a thin general syncytium incorporating few discrete cellular components, several of which share membrane continuity with the general syncytium by distinctive plug junctions. The general syncytium, supported by a thin collagenous mesolamella, is specialized regionally as dermal membrane, gastral membrane, peripheral trabecular strands, and primary reticulum (R1) of flagellated chambers. Extensions of the syncytium, which lack mesolamella support, form the distinctive secondary reticulum (R2) inside chambers and a newly discovered structure, the inner membrane, which occupies the central region of flagellated chambers. The choanosyncytia are enucleate networks of collar bodies and stolons embedded in R1 and plugged to R1 and choanoblasts. The discrete cell population consists of choanoblasts and archeocytes located in the thin mesohyle space and plugged to syncytial elements, cystencytes and vacuolar cells also located in the mesohyle but lacking plug connections, and granular cells emergent on R1 and apparently not bearing plug connections. The status of scleroblast syncytia has not been resolved. Large populations of rod-shaped bacteria occupy the mesohyle space; intracellular ovoid bodies, possible symbiotic prokaryotes, are common in R1 and R2. The previously unknown inner membrane probably functions to control flagellar activity on a very localized scale and to accumulate and release egesta in packages.Abbreviations ac archeocyte congeries - ap apopyle - ar archeocyte - b bacterium - c collagen fibrils - cb collar body - cbl choanoblast - cbs collar body socket - ch choanosome - cm collar microvilli - co choanocyte collar - cr crystalloid - cs connecting strand - dm dermal membrane - dv debris vacuole - e exhalant opening - ex exhalant space - f flagellum - fn filamentous network - Gb Golgi body - gf glycocalyx filaments - gm gastral membrane - im inner membrane - is inhalant space - ml mesolamella - ms mesohyle space - mt mitochondrion - n1 nucleus of R1 - o ostium - ob ovoid body - os osmiophilic body - pm plasma membrane - pr prosopyle - pt peripheral trabeculae - R1 primary reticulum - R2 secondary reticulum - s spicule space - ser smooth endoplasmic reticulum - ts trabecular strand     

The spermatogenesis ofHalichondria panicea (Porifera,Demospongiae)

Summary Spermatogenesis of the marine spongeHalichondria panicea begins with the break up of choanocyte chambers, choanocytes constituting the origin of spermatogonia. The transition from choanocytes to spermatogonia is direct, without cell division. Already the spermatogonia are flagellated. The ensuing large aggregates of spermatogonia are enclosed by spermatocyst-building cells. Further development takes place within the spermatocysts, mostly arranged in fields which, however, lack any developmental gradient. Within a single spermatocyst development is mostly synchronous. Spermatogonia transform into first order spermatocytes directly. The transition from spermatid to spermatozoon is characterized by an unusual prolongation of the chromatin, often resulting in a helical form of the chromosome material and a strong enlargement of the mitochondria which align with the nucleus, leading to an irregular shape of the spermatozoon. Another exceptional feature is the virtual absence of a Golgi apparatus during all stages of spermatogenesis. TheH. panicea investigated here contained only male reproductive elements, thus appear to be gonochorists. Some features of the spermatogenesis ofH. panicea, such as dissolving choanocyte chambers, the enclosure of spermatogonia by spermatocyst-building cells and the formation of a synaptonemal complex in first order spermatocytes occur in other sponge species as well; however, the early presence of flagella in spermatogonia, the absence of the Golgi apparatus and the later irregular development of nuclei, mitochondria and the spermatozoa themselves represent features hitherto not observed in sponges.