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.     

“Candidatus Paraholospora nucleivisitans”, an intracellular bacterium in Paramecium sexaurelia shuttles between the cytoplasm and the nucleus of its host

An intracellular bacterium was discovered in two isolates of Paramecium sexaurelia from an aquarium with tropical fish in Münster (Germany) and from a pond in the Wilhelma zoological–botanical garden, Stuttgart (Germany). The bacteria were regularly observed in the cytoplasm of the host, but on some occasions they were found in the macronucleus of the host cell. In these cases, only a few, if any, bacteria were observed remaining in the cytoplasm. The bacterium was not infectious to P. sexaurelia or other species of Paramecium and appeared to be an obligate intracellular bacterium, while bacteria-free host cells were completely viable. The fluorescence in situ hybridisation (FISH) and comparative 16SrDNA sequence analyses showed that the bacterium belonged to a new genus, and was most closely, yet quite distantly, related to Holospora obtusa. In spite of this relationship, the new bacteria differed from Holospora by at least two biological features. Whereas all Holospora species reside exclusively in the nuclei of various species of Paramecium and show a life cycle with a morphologically distinct infectious form, for the new bacterium no infectious form and no life cycle have been observed. For the new bacterium, the name Candidatus Paraholospora nucleivisitans is suggested. The host P. sexaurelia is usually known from tropical and subtropical areas and is not a species typically found in Germany and central Europe. Possibly, it had been taken to Germany with fish or plants from tropical or subtropical waters. Candidatus Paraholospora nucleivisitans may therefore be regarded as an intracellular neobacterium for Germany.     

Assessment of bacterial endosymbiont diversity in Otiorhynchusspp. (Coleoptera: Curculionidae) larvae using a multitag 454 pyrosequencing approach

Background

Weevils of the genus Otiorhynchus are regarded as devastating pests in a wide variety of horticultural crops worldwide. So far, little is known on the presence of endosymbionts in Otiorhynchus spp.. Investigation of endosymbiosis in this genus may help to understand the evolution of different reproductive strategies in these weevils (parthenogenesis or sexual reproduction), host-symbiont interactions, and may provide a future basis for novel pest management strategy development. Here, we used a multitag 454 pyrosequencing approach to assess the bacterial endosymbiont diversity in larvae of four economically important Otiorhynchus species.

Results

High-throughput tag-encoded FLX amplicon pyrosequencing of a bacterial 16S rDNA fragment was used to characterise bacterial communities associated with different Otiorhynchus spp. larvae. By sequencing a total of ~48,000 PCR amplicons, we identified 49 different operational taxonomic units (OTUs) as bacterial endosymbionts in the four studied Otiorhynchus species. More than 90% of all sequence reads belonged either to the genus Rickettsia or showed homology to the phylogenetic group of “Candidatus Blochmannia” and to endosymbionts of the lice Pedicinus obtusus and P. badii. By using specific primers for the genera Rickettsia and “Candidatus Blochmannia”, we identified a new phylogenetic clade of Rickettsia as well as “Candidatus Nardonella” endosymbionts in Otiorhynchus spp. which are closely related to “Candidatus Blochmannia” bacteria.

Conclusions

Here, we used multitag 454 pyrosequencing for assessment of insect endosymbiotic communities in weevils. As 454 pyrosequencing generates only quite short sequences, results of such studies can be regarded as a first step towards identifying respective endosymbiotic species in insects. In the second step of our study, we analysed sequences of specific gene regions for a more detailed phylogeny of selected endosymbiont genera. As a result we identified the presence of Rickettsia and “Candidatus Nardonella” endosymbionts in Otiorhynchus spp.. This knowledge is an important step in exploring bacteria-insect associations for potential use in insect pest control.

     

In situ pumping activity of the sponge Aplysina aerophoba, Nardo 1886

Aplysina aerophoba, Nardo 1886 is a common and well examined demosponge in the Mediterranean Sea. For sponges, as sessile, inner filter feeders, the most important function is pumping water through their canal system. This flow through is actively generated, supplies the sponge with oxygen and food particles and washes out waste products. Oxygen measurements ex situ, as a given example, show high oxygen saturations inside pumping sponges and oxygen depletion in non-pumping sponges. Thus, the oxygen situation within the sponge, like the food particle supply, waste washout and others, are directly related to its active pumping. To learn more about the poriferan function, it is important to know more about its undisturbed in situ pumping activity and consequently about the correlated conditions inside the sponge body.We conducted a tracer experiment in situ and tested sponge activity in terms of active pumping. Our technique excluded stress and disturbance to the sponges, hence minimizing experimental artefacts. The results show Aplysina aerophoba to be permanently pumping, which is implying a permanent supply of oxygen and of course food particles as well as a permanent washout of waste products and a permanent, presumably high energy consumption.We therefore conclude, that Aplysina aerophoba is always well supplied with oxygen, and that tissue anoxia or anaerobic metabolisms are of no significant importance in this sponge species. This fact of a permanent flow through in sponges will have to be taken into account for past and future hypotheses on the physiology of the sponge-microbial systems.     

AFM imaging of extracellular polymer release by marine diatom Cylindrotheca closterium (Ehrenberg) Reiman & J.C. Lewin

Extracellular polysaccharide production by marine diatoms is a significant route by which photosynthetically produced organic carbon enters the trophic web and may influence the physical environment in the sea. This study highlights the capacity of atomic force microscopy (AFM) for investigating diatom extracellular polysaccharides with a subnanometer resolution. Here we address a ubiquitous marine diatom Cylindrotheca closterium, isolated from the northern Adriatic Sea, and its extracellular polymeric substance (EPS) at a single cell level. We applied a simple procedure for AFM imaging of diatom cells on mica under ambient conditions (in air) to achieve visualization of their EPS with molecular resolution. The EPS represents a web of polysaccharide fibrils with two types of cross-linking: fibrils association forming junction zones and fibril-globule interconnections with globules connecting two or more fibrils. The fibril heights were 0.4-2.6 nm while globules height was in the range of 3-12 nm. Polymer networks of native gel samples from the Northern Adriatic and the network formed by polysaccharides extracted from the C. closterium culture share the same features regarding the fibril heights, pore openings and the mode of fibril association, proving that the macroscopic gel phase in the Northern Adriatic can be formed directly by the self-assembly of diatom released polysaccharide fibrils.     

Assessment of bacterial endosymbiont diversity in <Emphasis Type="Italic">Otiorhynchus</Emphasis>spp. (Coleoptera: Curculionidae) larvae using a multitag 454 pyrosequencing approach

BACKGROUND: Weevils of the genus Otiorhynchus are regarded as devastating pests in a wide variety of horticultural crops worldwide. So far, little is known on the presence of endosymbionts in Otiorhynchus spp.. Investigation of endosymbiosis in this genus may help to understand the evolution of different reproductive strategies in these weevils (parthenogenesis or sexual reproduction), host-symbiont interactions, and may provide a future basis for novel pest management strategy development. Here, we used a multitag 454 pyrosequencing approach to assess the bacterial endosymbiont diversity in larvae of four economically important Otiorhynchus species. RESULTS: High-throughput tag-encoded FLX amplicon pyrosequencing of a bacterial 16S rDNA fragment was used to characterise bacterial communities associated with different Otiorhynchus spp. larvae. By sequencing a total of ~48,000 PCR amplicons, we identified 49 different operational taxonomic units (OTUs) as bacterial endosymbionts in the four studied Otiorhynchus species. More than 90% of all sequence reads belonged either to the genus Rickettsia or showed homology to the phylogenetic group of "Candidatus Blochmannia" and to endosymbionts of the lice Pedicinus obtusus and P. badii. By using specific primers for the genera Rickettsia and "Candidatus Blochmannia", we identified a new phylogenetic clade of Rickettsia as well as "Candidatus Nardonella" endosymbionts in Otiorhynchus spp. which are closely related to "Candidatus Blochmannia" bacteria. CONCLUSIONS: Here, we used multitag 454 pyrosequencing for assessment of insect endosymbiotic communities in weevils. As 454 pyrosequencing generates only quite short sequences, results of such studies can be regarded as a first step towards identifying respective endosymbiotic species in insects. In the second step of our study, we analysed sequences of specific gene regions for a more detailed phylogeny of selected endosymbiont genera. As a result we identified the presence of Rickettsia and "Candidatus Nardonella" endosymbionts in Otiorhynchus spp.. This knowledge is an important step in exploring bacteria-insect associations for potential use in insect pest control.     

Sponges (Porifera) and eukaryotic, unicellular plankton: A case study on Aplysina aerophoba, Nardo 1886 in the Northern Adriatic

Sponges (Porifera), in general, are pumping water through their bodies. This water contains planktonic eukaryotic and procaryotic organisms as well as particulate and dissolved organic matter as potential food source. We analyzed the eukaryotic unicellular plankton fraction from water surrounding sponges of the species Aplysina aerophoba, Nardo 1886, from sponge tissue, as well as from water expelled from those sponges. We found sponges without any remnants of plankton in their tissue, as well as specimens which incorporated high numbers of remnants of organisms after planktonic “blooms”. In laboratory experiments, sponges were not showing any uptake of plankton from their surrounding water. Sponges are generally considered as inner filter feeders. However, our results indicate that eukaryotic unicellular plankton organisms are not the main food resource of the common sponge A. aerophoba. This raises the question if filter feeding is actually the main characteristic of the poriferan lifestyle.     

High decapod diversity revealed by minimal-invasive,short-term survey of Brijuni marine protected area

In our short-term, minimal-invasive decapod assessment of Brijuni marine protected area (MPA) we recorded 66 species belonging to 20 families. These represent a large part of the basic stock of decapods of the northern Adriatic, including some rare species. Moreover, we use species accumulation curves and extrapolation models to estimate total species richness, and to determine the effort needed for a viable decapod biodiversity assessment. Comparison with faunistic literature on Adriatic decapods indicates high species richness for Brijuni MPA.