Concurrent imaging of chlorophyll fluorescence,Chlorophyll a content and green fluorescent proteins‐like proteins of symbiotic cnidarians

Research on photosynthetic cnidarians has been mainly focused on the symbiosis established between the cnidarian host and its dinoflagellates endosymbionts from genus Symbiodinium. Despite the potential of imaging techniques for assessing the spatial distribution of key parameters of cnidarian photobiology, such as photochemical activity, chlorophyll a content or green fluorescent proteins (GFPs), to our best knowledge, no study has ever attempted to simultaneous map these three features. In this study, we developed a modified imaging pulse amplitude fluorometer by applying excitation light of different wavelengths and selectively detecting short spectral bands through bandpass filters. The imaging system was used to sequentially excite and quantify chlorophyll variable fluorescence (maximum quantum yield of photosystem II, F_v/F_m), Chl a content (normalized difference vegetation index) and relative content of GFPs. The spatial distribution of these photophysiological parameters was mapped both horizontally, across the surface of the soft corals Sarcophyton cf. glaucum and Sinularia flexibilis and the zoanthid Protopalythoa sp., and vertically, throughout a vertical section of S. cf. glaucum. Results showed bleached areas within each individual coral colony and registered photophysiological changes with S. cf. glaucum tissue depth. Analysis of Protopalythoa sp. polyps’ expansion revealed differential surface patterns of NDVI and GFP concentration, and a negative relation between these latter parameters within each polyp. This novel non‐invasive approach allowed a high‐resolution characterization of the spatial relationship between these key parameters through the analysis of image information on a pixel‐by‐pixel basis, which has great potential for investigating the physiological state of symbiotic associations.     

Genetic structure of the tall sea pen Funiculina quadrangularis in NW Scottish sea lochs

The tall sea pen Funiculina quadrangularis has a patchy distribution around the UK, being found in sheltered waters below 20 m depth on the northwest coast of Scotland and Ireland. The limited distribution and sensitivity to bottom fishing activities make F. quadrangularis vulnerable to reduction in population numbers that may lead to genetic isolation and reduced diversity. Because of this vulnerability and low resilience to physical disturbance, the tall sea pen is recognised as a Species of Principle Importance under the Natural Environment and Rural Communities Act, 2006, UK. ( http://www.legislation.gov.uk/ukpga/2006/16/contents ) and is also on the Biodiversity Action Plan list of Priority Species for the UK. In the Mediterranean it is recognised as a sensitive and essential fish habitat because it forms habitat for several commercially important crustaceans. The aim of this study was to understand the current state of the genetic structure and gene flow of F. quadrangularis in areas of NW Scotland. We developed 10 microsatellite markers and used them to genotype 176 samples from four populations. Overall, our results suggest that there is high genetic diversity and high gene flow between colonies of F. quadrangularis in and among locations in Loch Linnhe and Loch Duich. As a result of the high rates of gene flow, genetic differentiation between sites was low. This may provide resilience to human impacts if distant populations have a high connectivity. However, care must be taken, as small but significant isolation by distance was found between the most geographically distant sites and only a small part of the species range was examined in this study. The genetic tools developed here will provide a foundation for wider studies of this vulnerable species.     

Tracing dissolved organic matter cycling in the eastern boundary of the temperate North Atlantic using absorption and fluorescence spectroscopy

Apparent oxygen utilization (AOU), dissolved organic carbon (DOC), coloured dissolved organic matter (CDOM) absorption spectra, and CDOM fluorescence characteristic of aromatic amino acids (excitation/emission 280 nm/320 nm; F(280/320)) and marine-humic like substances (320 nm/410 nm; F(320/410)) were measured in full depth during a cruise in the temperate Eastern North Atlantic (ENA). An optimum multi-parameter (OMP) inverse method was run to calculate water mass proportion-weighted average (archetypal) concentrations of these chemical parameters for all water masses and samples. Archetypal concentrations retain the variability due to water mass mixing and basin scale mineralization from the water mass formation sites to the study area. Conversely, the difference between measured and archetypal concentrations, retain the variability due to dissimilarities in mineralization processes within the study area. Our analysis indicates that DOC supported 26±3% of the AOU in the dark temperate ENA and that basin scale processes occurring at and from the formation area of the water masses explained 63% of the total DOC variability. Our data also suggests that DOC remineralized at the basin scale was of lower molecular weight, and with a lower proportion of fluorescent aromatic amino acids than found within the study area. The relationship between the absorption coefficient at 254 nm (a_CDOM(254)) and AOU indicates that a_CDOM(254) was consumed during organic matter remineralization in the dark ocean, with 55% of the variability being explained by basin scale processes. The relationships of F(320/410) with AOU and DOC confirmed that marine humic-like substances are produced by microbial degradation processes, at a rate of 6.1±0.9×10⁻³ mg equivalents of QS mol AOU⁻¹. Our results also indicate that basin-scale remineralization processes account for 85% of the total variability of F(320/410), emphasizing that large scale processes control the formation of humic-like substance in the dark ENA.     

Effects of the oophagous bivalve Acesta oophaga on the morphology and fecundity of its deep‐sea tubeworm host,Lamellibrachia luymesi

The deep‐sea bivalve Acesta oophaga lives attached to the anterior end of the vestimentiferan tubeworm, Lamellibrachia luymesi, at cold methane seeps. The bivalve is found almost exclusively on female tubeworms, where it consumes the lipid‐rich eggs of L. luymesi that are spawned year round (Biological Bulletin, 209, 2005, 87). It is apparent that A. oophaga benefits directly from this close association, but the consequences for the tubeworm host may be more complicated than just a simple predator–prey interaction. Since A. oophaga completely surrounds the tube opening and plume of the worm, it is likely that its presence would limit oxygen uptake by L. luymesi, thereby inhibiting worm growth and reproduction. We hypothesized that occupied tubeworms would compensate for this by growing larger plumes for oxygen uptake. To explore the effects of bivalve presence/absence on female tubeworms, several morphological features, including body size, plume length, tube diameter, and tube segment length, as well as instantaneous fecundity, were compared. Results suggest that the mere presence of A. oophaga has a significant impact on the morphology of its host worm, as all measures of worm size, except for tube segment length, were significantly greater with clams present. Additionally, instantaneous fecundity was 3.5 times higher in occupied worms, implying that tubeworms are not oxygen‐deprived or energy limited as a result of bivalve presence. Our findings suggest that the association between these two deep‐sea organisms may be a more complex form of symbiosis than the simple predator–prey relationship, as previously thought.     

Interannual variability in the biochemical composition of newly hatched larvae of the spider crab Maja brachydactyla (Decapoda,Majidae)

The spider crab Maja brachydactyla is an important commercial species in Europe and supports intensive fisheries in the NE Atlantic. A field survey was performed to assess long‐term and consecutive interannual (2005–2010) variation of the biochemical composition of newly hatched larvae of M. brachydactyla. Larval biochemical profiles differed significantly among years, with pronounced differences being recorded in 2010. Differences among batches of newly hatched larvae were mainly explained by the contribution of triacylglycerols and, to a lesser degree, by protein and lipid content. The use of different nutrition indices is discussed. The biochemical composition of newly hatched larvae from M. brachydactyla was highly variable, even though surveyed broodstock was obtained from the same local population and was always sampled during the same season. The unpredictable biochemical profiles of newly hatched larvae may condition their survival and recruitment. This largely overlooked aspect of larval variability should be considered in future fisheries management strategies and captive production of marine organisms that still rely on wild seeds.     

Closely related thyasirid bivalves associate with multiple symbiont phylotypes

Species of thyasirid bivalves are considered to be representative of early stages of chemosymbiosis, given that bacterial symbionts occur outside gill epithelial cells, vary among species in their abundance and nutritional importance, and are environmentally acquired. For these reasons, assessing the extent of host–symbiont specificity in thyasirids could provide valuable information on the evolution of chemosymbiosis. We show that individuals of two closely related and sympatric Thyasira cf. gouldi operational taxonomic units collected from three sites in a fjord in Newfoundland, Canada, associate with one of three distinct, closely related symbiont phylotypes. While associations show some site‐specificity, there is flexibility in host–symbiont pairings within the fjord, further supporting an early and relatively unspecific stage of chemosymbiosis in this family. Morphologic differences observed both within and among symbiont phylotypes suggest physiologic variation, possibly induced by small‐scale differences in sedimentary conditions.     

Enhalus acoroides responses to experimental shoot density reductions in Haad Chao Mai National Park,Trang Province,Thailand

The effect of self‐shading and competition for light in the seagrass Enhalus acoroides were investigated with a density reduction experiment in Haad Chao Mai National Park, Trang Province, Thailand. The study was carried out in a monospecific meadow with a natural density of 141.0 ± 8.7 shoots·m^?2. The intent was to determine the response of E. acoroides beds to loss of shoots and thinning, which often occur during typhoons and severe storm activity. Permanent quadrats were manipulated by clipping the seagrass shoots to 140, 72, 36 and 16 shoots·m^?2, to yield natural, 50%, 25% and 10% densities, respectively. Reducing shoot density in E. acoroides increased underwater light intensity below the canopy, generating increased leaf surface area and shoot weight. Seagrass leaf width, growth rate, and number of leaves per shoot also increased with greater light. The extent of flowering varied among treatments with no consistent trend. Our results demonstrate that increasing the available light to E. acoroides produces an increasing leaf size response as self‐shading in the bed is reduced.     

Diel variation of benthic respiration in a coral reef sediment (Reunion Island,Indian Ocean)

Oxygen and total dissolved inorganic carbon (DIC) fluxes at the water–sediment interface were measured using benthic chambers to assess the short-term variations of community respiration (CR) in the back reef sediments of Reunion Island (Indian Ocean). Benthic CR had a daily cycle of minimal (6:00 AM) and maximal values (6:00 PM), showing increases of oxygen and DIC fluxes of 2.8- and 3.8-fold, respectively. Average CR values were observed at midday and midnight. The evolution of fluxes was positively related to oxygen concentration in ambient water, but not to temperature changes. In the study area, high daytime primary production augments the amount of energy available for community metabolism and increases benthic respiration. The benthic communities are therefore subjected to short-term variable environmental conditions with oxygen supersaturation during the day, and moderately hypoxic conditions at the end of the night.     

Methods for monitoring tidal flushing in large animal burrows in tropical mangrove swamps

The typically anaerobic nature of mangrove sediments provides significant challenges to the mangrove trees and biota inhabiting them. The burrowing activities and flow of water through the numerous and complex animal burrows perforating the sediments of mangroves have a major influence on the biogeochemistry of the sediments and are important to the enhancement of nutrient and oxygen exchange. Two new methods are presented for monitoring the tidal flushing of Sesarma messa and Alpheus cf macklay burrows in a Rhizophora stylosa mangrove forest – by measuring oxygen content of burrow water and by determining the change in fluorescence of a dye tracer through tidal inundation. A case study using the first of these showed oxygen consumption rates at the burrow wall deep within the burrow were found to be between 210 and 460 μmol O₂ m⁻² h⁻¹. The influx of oxygen during a flood tide was found to be significant and indicated that approximately 40% of the burrow water is flushed during a single tidal event. However, the high consumption rate of oxygen within the burrow resulted in the oxygen concentration remaining at or below one-third of the oxygen content of the flooding tidal water. A test application of the second method, using rhodamine dye as a tracer, indicated that the exchange of water between the burrow and the flooding tide was found to be in the order of 30% of the burrow volume. These new techniques provide a means to further study the nutrient exchange within these burrow systems and verify the initial findings that several tidal inundations are necessary to completely flush the burrows.     

Spatial and temporal variation in subtidal molluscan diversity amongst temperate estuarine habitats

Effective management of marine ecosystems is enhanced when detailed information on biodiversity is available. Key information to underpin management actions and conservation planning includes relationships between species assemblages and environmental gradients, and information on species distributions. We conducted a subtidal biodiversity assessment of surface‐dwelling subtidal molluscs in eight a priori defined habitat types using underwater visual censuses to quantitatively explore relationships between molluscan assemblages, and their correlation with benthic habitats and abiotic variables. In addition, variations in diversity were examined for two key habitat types (areas dominated by Dendronephthya australis and by filter feeders) over a period of 15 months to examine temporal change. We found that molluscs form distinct assemblages within subtidal habitats, but that assemblages within key habitats show inherent temporal variability. Regional (gamma) diversity of molluscs was found to result from a combination of: (i) within habitat alpha diversity, which increased with habitat complexity; (ii) between habitat beta diversity, with significant differences in molluscan assemblages amongst habitats with differing benthic growth, substrate type, and depth; and (iii) temporal beta diversity, with significant changes detected in molluscan assemblages over time. The results demonstrate how habitats and abiotic variables (principally depth and substrate type) combine to contribute to molluscan biodiversity in temperate estuaries, and illustrate the value of these factors as surrogates for surface‐dwelling subtidal molluscs in conservation planning.