Effects of Germanium (Ge) on the silica spicules of the marine sponge Suberites domuncula: Transformation of spicule type

Germanium (Ge), in the form of germanic acid, at a Ge/Si molar ratio of 1.0 inhibits gemmule development and silica deposition in the marine demosponge Suberites domuncula. Lower Ge/Si ratios inhibit the growth in length of the silica spicules (tylostyles) producing short structures, but with relatively normal morphology and close to normal width; spherical protuberances occasionally occur on these spicules. A few of the short spicules possess completely round rather than pointed tips. Many of the latter develop when Ge is added (pulsed) to growing animals, thus inducing a change in spicule type. These results indicate that the growth in length of the axial filament is more sensitive to Ge inhibition than is silica deposition and that pointed spicule tips normally develop because the growth of the axial filament at the spicule tip is more rapid than silica deposition. Newly formed spicules initiate silica deposition at the spicule head but the absence of Ge-induced bulbs as in freshwater spicules (oxeas) leaves open the question of whether there is a silicification center(s) present in Suberites tylostyles. The morphogenesis of freshwater oxeas and of marine tyolstyles appears fundamentally different-bidirectional growth in the former and unidirectional growth in the latter. X-ray analysis demonstrate relatively uniform Ge incorporation into the silica spicules with considerable variation from spicule to spicule in the incorporated level. Increased silicic acid concentration induces the formation of siliceous spheres, suggesting that the axial filament becomes prematurely encased in silica.     

Bone Marrow Biopsies with a Silverman Needle

Advantages and disadvantages of bone marrow biopsies taken from the posterior iliac crest using the Silverman needle were evaluated. Sixty-three biopsies were carried out in patients on whom previous aspirations of the bone marrow space were unsuccessful. Open surgical biopsy was thus avoided in all but one case. Only local anesthesia was required. In 12 cases the first attempt to obtain material with the Silverman needle was unsuccessful.Limitations of the method are largely due to crushing of the specimen so obtained, with polarization of nuclei simulating the appearance of fibroblasts seen in cases of myelosclerosis. At times marrow was completely expressed from the spongy bone, leaving a useless specimen of spicules. The quality of histological material is usually inferior to that yielded by successful aspiration.Despite limitations, the Silverman needle method is considered to be preferable to open biopsy in cases in which attempts at aspiration have failed.     

SPICULES IN SILURIAN TABULATE CORALS FROM CANADA, AND IMPLICATIONS FOR THEIR AFFINITIES

Abstract:  Specimens of Favosites from upper Llandovery strata of Anticosti Island show three types of calcite structures, herein interpreted as spicules, preserved within their calices and on top of the last tabula. This is stratigraphically younger material, some 50 m higher than fossils described two decades earlier, in which calcified polyps, each with 12 retracted tentacles, were noted. These more recently found structures show striking similarities in form and position to point, collaret and capstan spicules found in the soft tissues of modern pipe corals, i.e. the Octocorallia (Alcyonacea). Where preserved in a distinct pattern on top of the calcite tabulae, the spicular sclerites in Favosites occur in a particular sequence. Twelve individual, or sometimes six pairs of, triradiate point spicules have shrunk to a circlet near the middle of the calice (resting on the last, outermost, tabula). Surrounding the point spicules are 3–6 circlets of curved, usually perforated, lenticular collaret spicules; and surrounding these are scattered, much smaller, capstan spicules. The spicules display variability, probably ontogenetic, in their form and relative sizes; and they are more similar in form to calcareous spicules of alcyonacean corals than to those known from calcareous sponges. Structures with 12-fold radial symmetry in Heliolites, originally described by one of us as 'septal florets', consist of elements that are considered comparable with the point spicules found in Favosites . They have been recognized in ten species of Heliolites from Silurian (Wenlock–Ludlow) strata in the Canadian Arctic islands.     

Oldest record of freshwater sponges (Porifera: Spongillina) — spiculite finds in the Permo-Carboniferous of Europe

Up to now the oldest freshwater siliceous sponges are known from Jurassic deposits of Europe and North America. In this paper stratigraphically older Permo-Carboniferous freshwater siliceous sponges are reviewed/reported. Largely overlooked occurrences originate from the French Massif Central (Stefanian B), new discoveries come from the Saar-Nahe Basin (Stefanian C and Autunian) in south-west Germany. All Permo-Carboniferous finds originate from freshwater lake deposits situated hundreds of kilometers away from the sea. Preserved are monaxone spicules (oxeas) of various length/diameter ratios partly concentrated to spiculites. Gemmoscleres have not been found. A taxonomic classification on family level is not possible. Reasons for the immigration of marine euryhaline sponges into the freshwater might be the conquerence of the newly developed freshwater biotopes on Pangaea after the Variscan Orogeny. Possible migration routes are rivers connecting inland basins with the sea.     

Inferior ectopic pupil and typical ocular coloboma in RCS rats

Ocular coloboma is sometimes accompanied by corectopia in humans and therefore ectopic pupil may indicate ocular coloboma in experimental animals. The RCS strain of rats has a low incidence of microphthalmia. We found that inferior ectopic pupil is associated exclusively with small-sized eyes in this strain. The objective of the current study was to evaluate whether inferior ectopic pupil is associated with iridal coloboma and other types of ocular coloboma in RCS rats. Both eyes of RCS rats were examined clinically, and those with inferior ectopic pupils underwent morphologic and morphometric examinations. In a prenatal study, coronal serial sections of eyeballs from fetuses at gestational day 16.5 were examined by using light microscopy. Ectopic pupils in RCS rats were found exclusively in an inferior position, where the iris was shortened. Fundic examination revealed severe chorioretinal coloboma in all cases of inferior ectopic pupil. The morphologic characteristics closely resembled those of chorioretinal coloboma in humans. Histopathologic examination of primordia showed incomplete closure of the optic fissure in 4 eyeballs of RCS fetuses. Neither F(1) rats nor N(2) (progeny of RCS × BN matings) displayed any ocular anomalies, including ectopic pupils. The RCS strain is a suitable model for human ocular coloboma, and inferior ectopic pupil appears to be a strong indicator of ocular coloboma.     

Three new Procamallanus (Spirocamallanus) species from freshwater fishes in Mexico

The following 3 new species of Procamallanus (Spirocamallanus) are described from the intestines of freshwater fishes in Mexico, all belonging to the morphological group characterized by the presence of wide caudal alae, 3 pairs of subventral preanal papillae, and unequal spicules in the male: Procamallanus (Spirocamallanus) jaliscensis n. sp. (type host: Agonostomus monticola) and Procamallanus (Spirocamallanus) gobiomori n. sp. (hosts: Gobiomorus maculatus [type host], Gobiomorus polylepis and Eleotris picta) from 2 rivers in Jalisco State, western Mexico, and Procamallanus (Spirocamallanus) mexicanus n. sp. (type host: Cichlasoma geddesi) from Xalapa District, Veracruz State (Gulf of Mexico region), southeastern Mexico. Procamallanus jaliscensis is characterized by the length of the spicules (606-900 microm and 282-354 microm), number (15-16) of spiral ridges in the buccal capsule, and the digit-like protrusion with 1 terminal cuticular spike on the female tail; P. mexicanus by the length of the spicules (456-480 microm and 231-233 microm), number (10-12) of spiral ridges in the capsule, and the shape of the female tail (conical with a suddenly narrowed distal part, without any terminal spikes); and P. gobiomori by the length of spicules (318-348 microm and 156-192 microm), number (8-10) of spiral ridges and by the digit-like protrusion with 2 terminal cuticular spikes on the female tail.     

Raphidascaroides brasiliensis n. sp. (Nematoda: Anisakidae), an intestinal parasite of the thorny catfish Pterodoras granulosus from Amazonia,Brazil

A new anisakid nematode, Raphidascaroides brasiliensis n. sp., is described from the intestine of the freshwater thorny catfish Pterodoras granulosus (Valenciennes) (Doradidae, Siluriformes) from Amazonia (Manaus), Brazil. It is characterised mainly by the smooth, almost rounded tail tip in both sexes, the length of the spicules (0.952–1.183 mm) and by the number and arrangement of the caudal papillae (24–34 pre-anal, 1 adanal and 5 postanal pairs and 1 median pre-anal papilla) in the male. It is the first Raphidascaroides species described from South America and the second species of this genus reported from a freshwater fish.     

Spicules of hexactinellid sponges (Hexactinellida: Porifera) as natural composite materials

This paper reviews studies on the hexactinellid glass sponges (Hexactinellida: Porifera) that have organic silica spicules. According to its physical properties (microdensity, Young’s modulus, and light transmission), the material of the spicules is similar to amorphous silica; however, sponge spicules are birefringent, which suggests that they have a highly ordered crystal-like nature. Mineralized remnants of siliceous spicules composed of chemically inert materials are preserved in sedimentary rocks and provide evidence of the ecological state of the ancient biosphere. Sponges occur in waters with low temperatures; therefore, they grow very slowly and live for hundreds of years. The organic silica spicules exhibit the capacity for triboluminescence. The generated light emission may be used by symbiotic bacteria on the spicule surface.     

Comments on a skeleton design paradigm for a demosponge

The ball-shaped marine sponge Cinachyrellalevantinensis is 3-5 cm in diameter. It filters large quantities of seawater for feeding. Sponges contain numerous, hydrated, brittle amorphous SiO? spicules of several types that form 70-80% by weight of the sponge. We performed mechanical tests to determine the functionality of the sponge skeleton. The potential effect of habitat on skeleton properties was investigated by comparing sponges from 0.5 m and 30 m depth. We determined how spicules contribute to maintaining the strength and macroscopic structural integrity of a sponge, and studied their deformation mechanisms under external loading, and their microscopic design parameters. Compression tests of cylindrical samples cut from sponges revealed their macroscopic deformation mechanisms. Experiments solely with the organic material (following spicules dissolution) revealed the contribution of the spicules to the load carrying capacity and structural integrity of the sponge. Cantilever bending tests of anchored spicules determined the strength of individual spicules, the sponge's main skeletal elements. As the strength of brittle spicules is statistical in nature, we used Weibull Statistics to define their strength and evaluate their Young's modulus. Shallow and deep-water sponges did not differ significantly neither in response to compression, nor in spicule strength under bending and tension. Spicule weight fraction within a sponge was significantly higher in shallow-water individuals. We conclude that the structural integrity and strength of this sponge's skeleton is derived from its low-strength, small spicules, produced by a cost-effective process. The operating deformation of the spicules (bending) and their design parameters make them highly efficient.     

Dynamics of spicule production in the marine sponge <Emphasis Type="Italic">Hymeniacidon perlevis</Emphasis> during in vitro cell culture and seasonal development in the field

To characterize the formation of silica spicules, the dynamics of spiculogenesis of an intertidal marine sponge Hymeniacidon perlevis (Montagu 1818) (Porifera: Demospongiae) were investigated by measuring the gene expression of silicatein (the enzyme responsible for spicule silicification) and the dimensional changes of spicules during the developmental process of individual sponges and in cell cultures of primmorphs of archaeocyte-dominant cell populations. The different developmental stages of spicules were documented by time-lapse microscopy and observed by transmission electron microscopy during a 1-month culture period. During its annual life cycle, H. perlevis has four different developmental stages: dormancy, resuscitation, bloom, and decline. Field-grown individual sponge samples at different stages were collected over 7 months (March to September 2005). The dimensions of the silica spicules from these samples were microscopically measured and statistically analyzed. This analysis and the material properties of the spicules allowed them to be classified into four groups representing the different developmental stages of spiculogenesis. Silicatein expression in the bloom stage was more than 100 times higher than that in the other stages and was correlated with the spicule developmental stage. The trend of spicule formation in field-grown sponges was consistent with the trend in cell culture. A new parameter, the maturation degree (MD) of spicules (defined as the ratio of actual to theoretical silica deposition of mature spicules), was introduced to quantify spicule development. Silica spiculogenesis during H. perlevis development was delineated by comparing MD and silicatein expression.     

The cap-shaped Cambrian fossil Maikhanella and the relationship between coeloscleritophorans and molluscs

The recent discovery of large shells. growing by accretion, in the scleritome of the coeloscleritophoran Ha lkieria Poulsen, 1967, prompts the reconsideration of a number of cap-shaped shells in the Lower Cambrian. The scaly shells of Maikhanella Zhegallo, 1982. from Mongolia provide a good starting point, mce they occur together with spicules of siphogonuchitid type and appear to be composed of merged spicules. The shells are phosphatired and consist of two main elements: spicules, typically filled with phosphate. and an intermediate matrix. The associated loose spicules belong to forms that have served as the basis for erection of the genera Siphogonuchites Qian. 1977, and Lopochites Qian, 1977. Maikhanella is prohdbly a junior synonym of one or both of the latter two genera, but taxonomic revision is suspended until the type material of these siphogonuchitid genera has been restudied. and all three genera are left as sciotaxa. Various cap-shaped shells and plates in the Chinese Meishucunian are reinterpreted as shells belonging to coeloscleritophornn scleritoines. Maikhanella shells were formed through the embedding of spicules in secondary calcareous shell zubstance. The skeletogenesis has several similarities with that of molluscs, and together with the polyplacophoran-like features of the Halkieria scleritome this forces a reconsideration of the phylogenetic relationships between coeloscleritophorans and molluscs     

Hysterothylacium patagonense n. sp. (Nematoda: Anisakidae) from freshwater fishes in Patagonia,Argentina, with a key to the species of Hysterothylacium in American freshwater fishes

A new anisakid nematode, Hysterothylacium patagonense n. sp., is described from the intestine of the temperate bass Percichthys trucha (Cuvier & Valenciennes), a freshwater fish from Lake Aluminé, Patagonia, Argentina. It is characterised mainly by the absence of lateral alae, the length ratio of the caecum and ventricular appendix (1:0.9–1.8), the length of the spicules (0.952–1.292 mm) and the presence of minute spines on the tip of the tail. In the same locality and in Lake Huechulafquen, advanced third-stage and fourth-stage larvae of this parasite were also recorded from the digestive tract of the introduced salmonids, Salvelinus fontinalis (Mitchell), Oncorhynchus mykiss (Walbaum) and Salmo trutta L. This is the second Hysterothylacium species described from freshwater fishes in South America. A key to Hysterothylacium species parasitising American freshwater fishes is provided.     

Expression of silicatein in spicules from the Baikalian sponge Lubomirskia baicalensis

Lake Baikal harbors the largest diversity of sponge species [phylum Porifera] among all freshwater biotopes. The abundantly occurring species Lubomirskia baicalensis was used to study the seasonal silicatein metabolism; the spicules of this species have an unusually thick axial filament, consisting of silicatein, which remains constant in diameter during their growth. In the course of maturation, the size of the silicic acid shell grows, until the final diameter of the spicules of about 8 microm is reached. The seasonal content of silicatein was assessed by use of antibodies raised against silicatein; they stained specifically the axial filaments. In addition we determined, by application of the enzyme-linked immunosorbent assay system, that the proteinaceous content of the spicules, the silicatein, increases from spring to late summer by 8-fold. As molecular markers to quantify the seasonal changes in expression levels of genes coding for proteins/enzymes, the genes for the calumenin-like protein and the kinesin-related protein, were selected. The expression of calumenin-like gene, involved in the intracellular signaling, is highest during September, whereas the expression of the kinesin-related protein does not change during the annual course. These results suggest that the highest metabolic activity of L. baicalensis occurs in late summer (September), in parallel with the highest accumulation of silicatein, a structural protein/enzyme of the spicules.     

Light inside sponges

Sponges are the most basal metazoan organisms. As sessile filter feeders in marine or freshwater habitats, they often live in close association with phototrophic microorganisms. Active photosynthesis by the associated microorganisms has been believed to be restricted to the outer tissue portion of the sponge hosts. However, phototrophic microorganisms have also been detected in deeper tissue regions. In many cases they are found around spicules, siliceous skelettal elements of demosponges and hexactinellids. The finding of phototrophic organisms seemingly assembled around spicules led to the hypothesis of a siliceous light transmission system in sponges. The principle ability to conduct light was already shown for sponge derived, explanted spicules. However it was not shown until now, that in deed sponges have a light transmission system, and can harbour photosynthetically active microorganisms in deeper tissue regions.Here we show for the first time, that, as hypothesized 13 year ago, sponge spicules in living specimens transmit light into deeper tissue regions. Our results demonstrate that in opposite to the actual opinion, photosynthetically active microorganisms can also live in deeper tissue regions, and not only directly beneath the surface, when a light transmission system (spicules) is present.Our results show the possibility of massive or globular sponges being supplied with photosynthetic products or pathways throughout their whole body, implying not only a more important role of these endobioses. Our findings also elucidate the in-situ function of a recently more and more interesting biomaterial, which is unique not only for its mechanical, electrical and optical properties. Biosilica is of special interest for the possibility to produce it enzymatically under environmental conditions.     

Spicule structure and affinities of the Late Ordovician hexactinellid‐like sponge Cyathophycus loydelli from the Llanfawr Mudstones Lagerstätte,Wales

Exceptionally well‐preserved specimens of the reticulosan sponge Cyathophycus loydelli from the Sandbian (Late Ordovician) Llanfawr Mudstones Formation of Llandrindod, Waes, UK, have been examined using scanning electron microscopy (SEM). The specimens include exquisitely detailed pyritized spicules, and granular pyritization of surrounding soft tissues. Spicules frequently show axial canals of diameter similar to those of modern siliceous sponges, with hexagonal symmetry typical of modern demosponges rather than hexactinellids. In one case, the axial filament is also preserved. The largest spicules (ray diameter >20 μm) show a complex structure, with a laminar external region similar to that of the extant hexactinellid Monorhaphis. Some spicules preserve sub‐micron detail of the spicule surface, resembling the reticulate collagenous sheath of Monorhaphis. The hexagonal symmetry of the canal confirms that at least some Reticulosa are not crown‐group hexactinellids, but stem‐group Hexactinellida or Demospongea, or stem‐group Silicea. This suggests that a square canal is a sufficient diagnostic feature of total‐group Hexactinellida, but that hexagonal canals were more widely distributed among Early Silicea and were probably not restricted to demosponges. Alternatively, comparison with the structure of modern verongiid fibres suggests that these may be homologous with the outer layers of Cyathophycus spicules, and Cyathophycus may instead be a stem‐group demosponge. The preserved detail of the surface layer shows that pyritization can preserve certain material with extraordinarily fine resolution.     

Peculiar digestion patterns of sponge-associated zoochlorellae in the caddisfly Ceraclea fulva

The caddisfly Ceraclea fulva feeds exclusively on the freshwater sponge Ephydatia fluviatilis. Sponge spicules are accumulated in the insect midgut and arranged perpendicularly to the longitudinal axis of its gut. The peritrophic membrane of the midgut is so thick that it prevents spicules from damaging the epithelium during their transit. The digestion process of the endocellular zoochlorellae, which are vehiculated by the sponge cells, was examined by transmission electron microscopy (TEM). Zoochlorellae were seen in the midgut lumen, close to the peritrophic membrane and in the underlying space. Discrete algal cells became evident in tight apposition to the brush border of the midgut cells and were enveloped by the microvilli. Digestion progressed to the final transformation of the organism into membrane-delimited vacuoles.     

Effects of germanium on the morphology of silica deposition in a freshwater sponge

The effect of germanium on the secretion of siliceous spicules by the freshwater sponge Spongilla lacustris was investigated by exposing germinating and hatching gemmules to varying concentrations of germanium (Ge) in the presence of silicon (Si). Results were analyzed quantitatively and qualitatively and demonstrate that a [Ge]/[Si] (= molar ratio) of 1.0 completely inhibits silicon deposition. Intermediate ratios (0.5, 0.1, 0.01) which are permissive to spicule appearance result in fewer, shorter, and thinner spicules, in proportionately fewer microscleres, and in short bulbous megascleres. The size of the bulb increases with increasing [Ge]/[Si], while the length of the bulbous megascleres decreases with increasing [Ge]/[Si]. Microscleres do not demonstrate these graded responses suggesting that they are secreted in an all or none manner. Swellings produced in pond water and bulbs produced in germanium appear to decrease in size with time indicating a spreading of the accumulated silica. The effect of germanium on spicule secretion can be partially explained by its ability to uncouple the growth in length of the axial filament from the growth of the surrounding silicalemma. Under these conditions excess silicalemma is produced in which silica accumulates as bulbs in short spicules. Continuous exposure to Ge is necessary to produce this altered morphology. It is conjectured that the bulbs may be retained due to an inhibition of spreading. which in turn may be caused by the incorporation of germanium into the silica.