Audrey M. Pruski

Sensitivity to UV radiation in early life stages of the Mediterranean sea urchin Sphaerechinus granularis (Lamarck).

The sea urchin Sphaerechinus granularis was used to investigate the impact of relevant levels of UV-B radiation on the early life stages of a common Mediterranean free spawning benthic species. Sperm, eggs and embryos were exposed to a range of UV radiation doses. The resulting endpoints were evaluated in terms of fertilisation success, development and survival rates. Above a weighted UV radiation dose of 0.0029 kJ m(-2), fertilisation capability of irradiated sperm decreased rapidly. The exposure of the eggs to 0.0175 kJ m(-2) and more led to delayed and inhibited development with ensuing embryonic morphological abnormalities. One-day old larvae remained strongly sensitive to UV radiation as shown by the 50% decrease of the larval survival rate for a dose of 0.025 kJ m(-2) UVR. The elevated sensitivity of embryos to experimental UVR went along with a lack of significant amount of sunscreen compounds (e.g., mycosporine-like amino acids) in the eggs. The present results demonstrated that gamete viability and embryonic development may be significantly impaired by solar UV radiation in S. granularis, compromising in this way the reproduction of the species. Unless adaptive behavioural reproductive strategies exist, the influence of ambient UV radiation appears as a selective force for population dynamics of broadcast spawners in the shallow benthic Mediterranean environment.

River organic matter shapes microbial communities in the sediment of the Rhône prodelta

Microbial-driven organic matter (OM) degradation is a cornerstone of benthic community functioning, but little is known about the relation between OM and community composition. Here we use Rhône prodelta sediments to test the hypothesis that OM quality and source are fundamental structuring factors for bacterial communities in benthic environments. Sampling was performed on four occasions corresponding to contrasting river-flow regimes, and bacterial communities from seven different depths were analyzed by pyrosequencing of 16S rRNA gene amplicons. The sediment matrix was characterized using over 20 environmental variables including bulk parameters (for example, total nitrogen, carbon, OM, porosity and particle size), as well as parameters describing the OM quality and source (for example, pigments, total lipids and amino acids and δ13C), and molecular-level biomarkers like fatty acids. Our results show that the variance of the microbial community was best explained by δ13C values, indicative of the OM source, and the proportion of saturated or polyunsaturated fatty acids, describing OM lability. These parameters were traced back to seasonal differences in the river flow, delivering OM of different quality and origin, and were directly associated with several frequent bacterial operational taxonomic units. However, the contextual parameters, which explained at most 17% of the variance, were not always the key for understanding the community assembly. Co-occurrence and phylogenetic diversity analysis indicated that bacteria–bacteria interactions were also significant. In conclusion, the drivers structuring the microbial community changed with time but remain closely linked with the river OM input.

Ultraviolet radiation induces structural and chromatin damage in Mediterranean sea-urchin spermatozoa.

There is growing concern about the effects of enhanced levels of solar ultraviolet radiation on the living components of the biosphere (i.e. cancer, loss of biodiversity and productivity, etc.). In shallow coastal environments, many benthic species release their gametes directly in the water column where fertilisation occurs and the planktonic larvae remain for several weeks. Any effects on these early life stages could significantly impair reproductive input or alter the fitness of the community. The purpose of this paper is to provide new insights into the mechanisms of UV toxicity on sea-urchin spermatozoa in a cytological context, and to address the question of the potential ecological consequences of the damage. The Mediterranean sea-urchin Sphaerechinus granularis (Lamarck) was chosen as a model to study the effects of ecologically relevant doses of UV-R on the spermatozoa of marine invertebrates. Structural damage was visualised by use of transmission electron microscopy and the single-cell gel electrophoresis (Comet) assay was used to assess chromatin integrity in spermatozoa. The present results provide experimental evidence that irradiation with UV induces structural and chromatin damage in sea-urchin sperm. Almost 90% of spermatozoa exhibited morphological alterations and DNA strand breakage increased 2-fold. The observed alterations of the acrosome, plasma membrane and mitochondria can explain the concomitant impairment of fertilisation (23% decrease of fertilisation rate), which in turn may affect reproductive success. On the other hand, how DNA damage and fertilisation rate correlate remains unclear; however, when not repaired genetic lesions can lead to abnormal development and/or the transmission of heritable damage. The 3-fold decrease of the frequency of 2-celled embryos indicates a delay or inhibition of the first cell division, which may be ascribed to impairment of nuclear chromatin and/or other cellular targets.

Improved Comet assay for the assessment of UV genotoxicity in Mediterranean sea urchin eggs.

Gametes and embryos of broadcast spawners are exposed to a wide range of chemical and physical stressors which may alone, or in conjunction, have serious consequences on reproductive outcomes. In this study, two Mediterranean echinoid species, Paracentrotus lividus and Sphaerechinus granularis, were chosen as models to study the genotoxicity of UV radiation (UVR) on the eggs of broadcast‐spawning marine invertebrates. The single cell gel electrophoresis, or Comet assay, was successfully adapted to assess DNA strand breakage in sea urchin eggs. The results demonstrated that the genetic material of sea urchin eggs is susceptible to environmentally realistic UV exposure. The induction of DNA damage in the irradiated unfertilized eggs suggests that the previously described defense mechanisms in sea urchin eggs do not completely protect the eggs DNA against UV toxicity. Taken together, our results suggest that UV‐impairment of the genetic integrity of the eggs might have a role in postfertilization failures and abnormal embryonic development. Although both species were vulnerable to UVR, embryonic development was less dramatically impaired in P.Lividus. This observation supports the postulation that species inhabiting shallower environments possess more efficient mechanisms to overcome UV‐induced DNA alterations. The present demonstration of the utility and sensitivity of the Comet assay to evaluate DNA integrity in eggs from marine invertebrates opens new perspectives for monitoring the long‐term effects of environmental exposure on populations and for the routine screening of substances for genotoxicity in marine systems. Environ. Mol. Mutagen., 2008.

Temporal and spatial constraints on community assembly during microbial colonization of wood in seawater

Wood falls on the ocean floor form chemosynthetic ecosystems that remain poorly studied compared with features such as hydrothermal vents or whale falls. In particular, the microbes forming the base of this unique ecosystem are not well characterized and the ecology of communities is not known. Here we use wood as a model to study microorganisms that establish and maintain a chemosynthetic ecosystem. We conducted both aquaria and in situ deep-sea experiments to test how different environmental constraints structure the assembly of bacterial, archaeal and fungal communities. We also measured changes in wood lipid concentrations and monitored sulfide production as a way to detect potential microbial activity. We show that wood falls are dynamic ecosystems with high spatial and temporal community turnover, and that the patterns of microbial colonization change depending on the scale of observation. The most illustrative example was the difference observed between pine and oak wood community dynamics. In pine, communities changed spatially, with strong differences in community composition between wood microhabitats, whereas in oak, communities changed more significantly with time of incubation. Changes in community assembly were reflected by changes in phylogenetic diversity that could be interpreted as shifts between assemblies ruled by species sorting to assemblies structured by competitive exclusion. These ecological interactions followed the dynamics of the potential microbial metabolisms accompanying wood degradation in the sea. Our work showed that wood is a good model for creating and manipulating chemosynthetic ecosystems in the laboratory, and attracting not only typical chemosynthetic microbes but also emblematic macrofaunal species.

Can UV radiation affect benthic deposit-feeders through biochemical alteration of food resources? An experimental study with juveniles of the benthic polychaete Eupolymnia nebulosa.

The growth, tentacle development and feeding activity of the benthic polychaete Eupolymnia nebulosa were examined to determine whether UV might affect marine deposit-feeders indirectly through the modification of the nutritional quality of their resources. Since marine invertebrates have higher nutritional requirements during the period following settlement, we tested the effect of UV-altered phytodetritus on freshly settled juveniles of E. nebulosa. Phytodetritus was prepared from cultures of the diatom Skeletonema costatum either grown under or sheltered from UVB radiation. Sterol content of phytodetritus was unmodified by UV radiation. Conversely, phytodetritus was noticeably depleted in polyunsaturated fatty acids. Growth and tentacle development of juveniles fed on altered phytodetritus were reduced by 35% and 15% respectively, suggesting potential deficiencies in essential nutrients. In response to the lower quality of the phytodetritus, juveniles explored a wider area as they search for food, a strategy that could compensate for low food quality.