ALBERT

Description: Experiments are described to define further the fatty acid requirements of rainbow trout (Salmo gairdneri). In all cases, feeding semipurified diets containing no polyunsaturated fatty acids resulted in poor growth and feed conversion. Linolenic acid was superior to linoleic in stimulating growth and improving feed conversion. The requirement of linolenic acid (ω3 fatty acids) for rainbow trout is 1% of the diet or approximately 2.7% of the dietary calories. Essential fatty acid deficiency symptoms that were cured or prevented by linolenic acid included fin erosion, heart myopathy, and a shock syndrome. It is concluded that linolenic acid has an essential role in rainbow trout similar to that assigned to linoleic acid in man and higher animals.

Description: Work on the life histories of common antarctic benthic marine invertebrates over the past several decades demands a revision of several widely held paradigms. First, contrary to expectations derived from work on temperate species, there is little or no evidence for temperature adaptation with respect to reproduction (gametogenesis), devel? opment, and growth. It remains to be determined whether the slow rates of these processes reflect some inherent inability to adapt to low temperatures, or are a response to features of the antarctic marine environment not directly related to low temperature, such as low food resources. Secondly, contrary to the widely accepted opinion designated as "Thor- son's rule," pelagic development is common in many groups of shallow-water marine invertebrates. In fact in some groups, such as asteroids, pelagic development is as prevalent in McMurdo Sound, the southern-most open-water marine environment in the world, as in central California. In other taxonomic groups, especially gastropods, there does seem to be a genuine trend toward non-pelagic development from tropical to antarctic latitudes. Although this trend has been predicted by theoretical models, its underlying causes appear to be group specific rather than general. Thirdly, pelagic lecithotrophic development, often considered to be of negligible importance, occurs in many shallow-water antarctic marine macroinvertebrates. Pelagic lecithotrophy may be an adaptation to a combination of poor food conditions in antarctic waters most ofthe year and slow rates of development. Nevertheless, some of the most abundant and widespread antarctic marine invertebrates have pelagic planktotrophic larvae that take very long times to complete development to metamorphosis. These species are particularly prevalent in productive regions of shallow water (〈 30 m), which are frequently disturbed by anchor ice formation, and the production of numerous pelagic planktotrophic larvae may represent a strategy for colonization. Although planktotrophic larvae tend to be seasonal in occurrence, their production is not linked particularly closely to the mid-summer pulse of phytoplankton production. These larvae show no evidence of starvation, even during times when phytoplankton abundance is very low, and they may depend on unusual sources of food, such as bacteria. How they escape the selective conditions that apparently led to a predominance of non-feeding modes of development in antarctic marine invertebrates remains as a major challenge for antarctic marine biology.

Description: Nine semipurified diets containing protein/energy ratios that ranged from 73 to 162 mg protein/kilocalorie were fed to young rainbow trout for 18 weeks. A casein-gelatin (70–30) mixture and herring oil were each fed at three levels in a factorial type of experiment. Each of the casein-gelatin levels (36, 44 and 53%) was fed at each of the fat levels (8, 16 and 24%). Cornstarch was added at the expense of the casein-gelatin mix to adjust dietary protein levels. Caloric intake regulated feed consumption; and except for diets low in both fat and protein, no significant differences in weight gains were noted, although feed and energy conversions were markedly influenced. Higher protein/calorie ratios were positively correlated with liver size, level of liver sugars, percentage body fat, percentage body protein, and negatively correlated with percentage liver lipids, size of gastrointestinal tract and gain/gram ingested protein. These correlations were observed for similar protein/calorie ratios regardless of the dietary levels of protein and lipid.

Description: Mixing processes of reduced hydrothermal fluids with oxygenated seawater and fluid-rock reactions contribute to the chemical signatures of diffuse venting and likely determine the geochemical constraints on microbial life. We examined the influence of fluid chemistry on microbial diversity and activity by sampling diffuse fluids emanating through mussel beds at two contrasting hydrothermal vents. The H(2) concentration was very low at the basalt-hosted Clueless site, and mixing models suggest O(2) availability throughout much of the habitat. In contrast, effluents from the ultramafic-hosted Quest site were considerably enriched in H(2) , while O(2) is likely limited to the mussel layer. Only two different hydrogenase genes were identified in clone libraries from the H(2) -poor Clueless fluids, but these fluids exhibited the highest H(2) uptake rates in H(2) -spiked incubations (oxic conditions, at 18 °C). In contrast, a phylogenetically diverse H(2) -oxidizing potential was associated with distinct thermal conditions in the H(2) -rich Quest fluids, but under oxic conditions, H(2) uptake rates were extremely low. Significant stimulation of CO(2) fixation rates by H(2) addition was solely illustrated in Quest incubations (P-value 〈0.02), but only in conjunction with anoxic conditions (at 18 °C). We conclude that the factors contributing toward differences in the diversity and activity of H(2) oxidizers at these sites include H(2) and O(2) availability.

Description: We present high-resolution resistivity imaging of gas hydrate pipe-like structures, as derived from marine controlled-source electromagnetic (CSEM) inversions that combine towed and ocean-bottom electric field receiver data, acquired from the Nyegga region, offshore Norway. Two-dimensional CSEM inversions applied to the towed receiver data detected four new prominent vertical resistive features that are likely gas hydrate structures, located in proximity to a major gas hydrate pipe-like structure, known as the CNE03 pockmark. The resistivity model resulting from the CSEM data inversion resolved the CNE03 hydrate structure in high resolution, as inferred by comparison to seismically constrained inversions. Our results indicate that shallow gas hydrate vertical features can be delineated effectively by inverting both ocean-bottom and towed receiver CSEM data simultaneously. The approach applied here can be utilised to map and monitor seafloor mineralisation, freshwater reservoirs, CO2 sequestration sites and near-surface geothermal systems.

Description: Ocean acidification (OA) is increasing due to anthropogenic CO2 emissions and poses a threat to marine species and communities worldwide. To better project the effects of acidification on organisms’ health and persistence, an understanding is needed of the 1) mechanisms underlying developmental and physiological tolerance and 2) potential populations have for rapid evolutionary adaptation. This is especially challenging in nonmodel species where targeted assays of metabolism and stress physiology may not be available or economical for large-scale assessments of genetic constraints. We used mRNA sequencing and a quantitative genetics breeding design to study mechanisms underlying genetic variability and tolerance to decreased seawater pH (-0.4 pH units) in larvae of the sea urchin Strongylocentrotus droebachiensis. We used a gene ontology-based approach to integrate expression profiles into indirect measures of cellular and biochemical traits underlying variation in larval performance (i.e., growth rates). Molecular responses to OA were complex, involving changes to several functions such as growth rates, cell division, metabolism, and immune activities. Surprisingly, the magnitude of pH effects on molecular traits tended to be small relative to variation attributable to segregating functional genetic variation in this species. We discuss how the application of transcriptomics and quantitative genetics approaches across diverse species can enrich our understanding of the biological impacts of climate change.

Description: Barnacles of the genus Galkinius occupy a large spectrum of host corals, making it one of the least host-specific genera within the Pyrgomatidae. Molecular analyses show that within the genus Galkinius there are highly supported clades, suggesting that the genus Galkinius is a complex of evolutionarily significant units (ESUs). The morphology of the opercular valves has been used as the basis for the separation of species of Galkinius. In this study, morphological variability was found both between specimens within ESUs extracted from different host species and between specimens extracted from the same colony. Identifications based on the opercular valves cannot therefore be assigned to different species despite being genetically distinguishable. It is proposed that in many cases the differences between valve morphology of different species of Galkinius are the outcome of ontogeny. Allometric growth of the valves has resulted in differences in the proportions of the parts of the valve.

Description: Pseudovibrio is a marine bacterial genus members of which are predominantly isolated from sessile marine animals, and particularly sponges. It has been hypothesised that Pseudovibrio spp. form mutualistic relationships with their hosts. Here, we studied Pseudovibrio phylogeny and genetic adaptations that may play a role in host colonization by comparative genomics of 31 Pseudovibrio strains, including 25 sponge isolates. All genomes were highly similar in terms of encoded core metabolic pathways, albeit with substantial differences in overall gene content. Based on gene composition, Pseudovibrio spp. clustered by geographic region, indicating geographic speciation. Furthermore, the fact that isolates from the Mediterranean Sea clustered by sponge species suggested host-specific adaptation or colonization. Genome analyses suggest that Pseudovibrio hongkongensis UST20140214-015BT is only distantly related to other Pseudovibrio spp., thereby challenging its status as typical Pseudovibrio member. All Pseudovibrio genomes were found to encode numerous proteins with SEL1 and tetratricopeptide repeats, which have been suggested to play a role in host colonization. For evasion of the host immune system, Pseudovibrio spp. may depend on type III, IV and VI secretion systems that can inject effector molecules into eukaryotic cells. Furthermore, Pseudovibrio genomes carry on average seven secondary metabolite biosynthesis clusters, reinforcing the role of Pseudovibrio spp. as potential producers of novel bioactive compounds. Tropodithietic acid, bacteriocin and terpene biosynthesis clusters were highly conserved within the genus, suggesting an essential role in survival e.g. through growth inhibition of bacterial competitors. Taken together, these results support the hypothesis that Pseudovibrio spp. have mutualistic relations with sponges.

Description: We studied the effect of the quorum-sensing (QS) blockers 5-hydroxy-3[(1R)-1-hydroxypropyl]-4-methylfuran-2(5H)-one (FUR1), (5R)-3,4-dihydroxy-5-[(1S)-1,2-dihydroxyethyl]furan-2(5H)-one (FUR2) and triclosan (TRI) on the formation of bacterial biofilms, and the effect of these biofilms on the larval attachment of the polychaete Hydroides elegans and the bryozoan Bugula neritina. 14-day-old subtidal biofilms were harvested from artificial substrata and were allowed to develop in the laboratory with and without QS blockers. QS blockers inhibited the production of violacein by the QS reporter strain Chromobacterium violaceum CV026 and did not affect the metabolic activity of bacteria in multispecies biofilms. At a concentration of 10(-3) M all three tested compounds inhibited the establishment of microbial communities, but at one of 10(-4) M only FUR2 inhibited establishment. The tested QS blockers caused changes in bacterial density and bacterial community structure, as revealed by terminal restriction fragment length polymorphism and FISH. The groups most affected by QS blockers were Alphaproteobacteria, Gammaproteobacteria and the Cytophagales. Larvae of H. elegans and B. neritina avoided settling on biofilms that had developed in the presence of QS blockers. Our results suggest that QS blockers directly control the formation of multi-species biofilms, and indirectly - by means of biofilm properties - affect larval attachment on these modified biofilms.

Description: The macroalga Fucus vesiculosus carries a specific community of surface bacteria. To identify chemical compounds that possibly mediate abundance and community composition of algae-associated bacteria, we tested the effect of surface-available algal compounds on bacterial settlement and community composition under field conditions. Compounds on algal thalli were separated from the surface by extraction with organic solvents and investigated on growth inhibition and settlement of bacterial isolates. Based on in vitro data, partially purified extract fractions were then exposed to bacterial colonizers in vivo followed by bacterial enumeration and community analysis. The polar fraction of the algal surface extract revealed a significant profouling effect for Vibrionales, whereas the nonpolar fraction containing the xanthophyll pigment fucoxanthin and other unidentified nonpolar surface compounds revealed a significant 80% reduction of surface colonizing bacteria. The analysis of bacterial surface communities by 454 pyrosequencing demonstrated that the antifouling activity of nonpolar algal surface compounds was targeting the abundance of natural bacterial colonizers rather than the relative composition of bacterial members within the community. Moreover, the bacterial community composition on F.vesiculosus was markedly different from artificial control substrates and chemically manipulated experimental treatments, suggesting that other, nonextractable surface features and/or physical properties render algal-specific epiphytic bacterial communities.

Description: To understand how ocean acidification (OA) influences sediment microbial communities, naturally CO2-rich sites are increasingly being used as OA analogues. However, the characterization of these naturally CO2-rich sites is often limited to OA-related variables, neglecting additional environmental variables that may confound OA effects. Here, we used an extensive array of sediment and bottom water parameters to evaluate pH effects on sediment microbial communities at hydrothermal CO2seeps in Papua New Guinea. The geochemical composition of the sediment pore water showed variations in the hydrothermal signature at seep sites with comparable pH, allowing the identification of sites that may better represent future OA scenarios. At these sites, we detected a 60% shift in the microbial community composition compared with reference sites, mostly related to increases inChloroflexisequences. pH was among the factors significantly, yet not mainly, explaining changes in microbial community composition. pH variation may therefore often not be the primary cause of microbial changes when sampling is done along complex environmental gradients. Thus, we recommend an ecosystem approach when assessing OA effects on sediment microbial communities under natural conditions. This will enable a more reliable quantification of OA effects via a reduction of potential confounding effects

Description: Connectivity of pelagic, early life stages via transport by ocean currents may affect survival chances of offspring, recruitment success, and mixing of stocks across management units. Based on drift model studies, transport patterns of particles representing exogenously feeding cod larvae in the transition area between North Sea and Baltic were investigated to (i) determine long-term trends and variability in advective transport of larvae from spawning grounds to juvenile nursery areas, (ii) estimate the degree of exchange between different management areas, and (iii) compare the results with spatial distributions of juvenile cod. The transport of particles showed considerable intra- and interannual variability, but also some general patterns of retention within and dispersion to different management areas. Good spatial overlap of particle end positions, representing potential juvenile settlement areas, with observed distributions of juveniles in bottom trawl surveys suggests that the drift simulations provide reasonable estimates of early life stage connectivity between cod populations in the investigated areas. High exchange rates of particles between management areas of up to ca. 70% suggest that cod populations in the investigated areas are demographically correlated. Results are discussed in relation to their relevance for stock structure, fish stock assessment, and management.

Description: The diversity of Cyanobacteria in water and sediment samples from four representative sites of the Salar de Huasco was examined using denaturing gradient gel electrophoresis and analysis of clone libraries of 16S rRNA gene PCR products. Salar de Huasco is a high altitude (3800 m altitude) saline wetland located in the Chilean Altiplano. We analyzed samples from a tributary stream (H0) and three shallow lagoons (H1, H4, H6) that contrasted in their physicochemical conditions and associated biota. Seventy-eight phylotypes were identified in a total of 268 clonal sequences deriving from seven clone libraries of water and sediment samples. Oscillatoriales were frequently found in water samples from sites H0, H1 and H4 and in sediment samples from sites H1 and H4. Pleurocapsales were found only at site H0, while Chroococcales were recovered from sediment samples of sites H0 and H1, and from water samples of site H1. Nostocales were found in sediment samples from sites H1 and H4, and water samples from site H1 and were largely represented by sequences highly similar to Nodularia spumigena. We suggest that cyanobacterial communities from Salar de Huasco are unique - they include sequences related to others previously described from the Antarctic, along with others from diverse, but less extreme environments.

Description: As ecosystem engineers, seagrasses are angiosperms of paramount ecological importance in shallow shoreline habitats around the globe. Furthermore, the ancestors of independent seagrass lineages have secondarily returned into the sea in separate, independent evolutionary events. Thus, understanding the molecular adaptation of this clade not only makes significant contributions to the field of ecology, but also to principles of parallel evolution as well. With the use of Dr. Zompo, the first interactive seagrass sequence database presented here, new insights into the molecular adaptation of marine environments can be inferred. The database is based on a total of 14 597 ESTs obtained from two seagrass species, Zostera marina and Posidonia oceanica, which have been processed, assembled and comprehensively annotated. Dr. Zompo provides experimentalists with a broad foundation to build experiments and consider challenges associated with the investigation of this class of non-domesticated monocotyledon systems. Our database, based on the Ruby on Rails framework, is rich in features including the retrieval of experimentally determined heat-responsive transcripts, mining for molecular markers (SSRs and SNPs), and weighted key word searches that allow access to annotation gathered on several levels including Pfam domains, GeneOntology and KEGG pathways. Well established plant genome sites such as The Arabidopsis Information Resource (TAIR) and the Rice Genome Annotation Project are interfaced by Dr. Zompo. With this project, we have initialized a valuable resource for plant biologists in general and the seagrass community in particular. The database is expected to grow together with more data to come in the near future, particularly with the recent initiation of the Zostera genome sequencing project. The Dr. Zompo database is available at http://drzompo.uni-muenster.de/

Description: In this pilot study, we describe a high-pressure incubation system allowing multiple subsampling of a pressurized culture without decompression. The system was tested using one piezophilic (Photobacterium profundum), one piezotolerant (Colwellia maris) bacterial strain and a decompressed sample from the Mediterranean deep sea (3044 m) determining bacterial community composition, protein production (BPP) and cell multiplication rates (BCM) up to 27 MPa. The results showed elevation of BPP at high pressure was by a factor of 1.5 ± 1.4 and 3.9 ± 2.3 for P. profundum and C. maris, respectively, compared to ambient-pressure treatments and by a factor of 6.9 ± 3.8 fold in the field samples. In P. profundum and C. maris, BCM at high pressure was elevated (3.1 ± 1.5 and 2.9 ± 1.7 fold, respectively) compared to the ambient-pressure treatments. After 3 days of incubation at 27 MPa, the natural bacterial deep-sea community was dominated by one phylum of the genus Exiguobacterium, indicating the rapid selection of piezotolerant bacteria. In future studies, our novel incubation system could be part of an isopiestic pressure chain, allowing more accurate measurement of bacterial activity rates which is important both for modeling and for predicting the efficiency of the oceanic carbon pump.

Description: Legionella pneumophila is naturally found in fresh water were the bacteria parasitize within protozoa. It also survives planctonically in water or biofilms. Upon aerosol formation via man-made water systems, L. pneumophila can enter the human lung and cause a severe form of pneumonia, called Legionnaires’ disease. The pathogenesis of Legionnaires’ disease is largely due to the ability of L. pneumophila to invade and grow within macrophages. An important characteristic of the intracellular survival strategy is the replication within the host vacuole that does not fuse with endosomes or lysosomes. In recent times a great number of bacterial virulence factors which affect growth of L. pneumophila in both macrophages and protozoa have been identified. The ongoing Legionella genome project and the use of genetically tractable surrogate hosts are expected to significantly contribute to the understanding of bacterium–host interactions and the regulation of virulence traits during the infection cycle. Since person-to-person transmission of legionellosis has never been observed, the measures for disease prevention have concentrated on eliminating the pathogen from water supplies. In this respect detection and analysis of Legionella in complex environmental consortia become increasingly important. With the availability of new molecular tools this area of applied research has gained new momentum.

Description: Archaea associated with marine sponges are active and influence the nitrogen metabolism of sponges. However, we know little about their occurrence, specificity, and persistence. We aimed to elucidate the relative importance of host specificity and biogeographic background in shaping the symbiotic archaeal communities. We investigated these communities in sympatric sponges from the Mediterranean (Ircinia fasciculata and Ircinia oros, sampled in summer and winter) and from the Caribbean (Ircinia strobilina and Mycale laxissima). PCR cloning and sequencing of archaeal 16S rRNA and amoA genes showed that the archaeal community composition and structure were different from that in seawater and varied among sponge species. We found that the communities were dominated by ammonia-oxidizing archaea closely related to Nitrosopumilus. The community in M. laxissima differed from that in Ircinia spp., including the sympatric sponge I. strobilina; yet, geographical clusters within Ircinia spp. were observed. Whereas archaeal phylotypes in Ircinia spp. were persistent and belong to 'sponge-enriched' clusters, archaea in M. laxissima were closely related with those from diverse habitats (i.e. seawater and sediments). For all four sponge species, the expression of the archaeal amoA gene was confirmed. Our results indicate that host-specific processes, such as host ecological strategy and evolutionary history, control the sponge-archaeal communities.

Description: The ability of microorganisms to withstand long periods with extremely low energy input has gained increasing scientific attention in recent years. Starvation experiments in the laboratory have shown that a phylogenetically wide range of microorganisms evolve fitness-enhancing genetic traits within weeks of incubation under low-energy stress. Studies on natural environments that are cut off from new energy supplies over geologic time scales, such as deeply buried sediments, suggest that similar adaptations might mediate survival under energy limitation in the environment. Yet, the extent to which laboratory-based evidence of starvation survival in pure or mixed cultures can be extrapolated to sustained microbial ecosystems in nature remains unclear. In this review, we discuss past investigations on microbial energy requirements and adaptations to energy limitation, identify gaps in our current knowledge, and outline possible future foci of research on life under extreme energy limitation.

Description: The permafrost methane emission problem is the focus of attention on different climate models. Here, we present a mathematical model for permafrost lake methane emission and its influence on the climate system. We model this process using the theory of non-linear phase transitions. Further, we find that a climate catastrophe possibility depends on a value of feedback connecting the methane concentration in the atmosphere and temperature, and on the tundra permafrost methane pool.We note that the permafrost lake model that we developed for the methane emission positive feedback loop problem is a conceptual climate model.

Description: The GeoChip 4.2 gene array was employed to interrogate the microbial functional gene repertoire of sponges and seawater collected from the Red Sea and the Mediterranean. Complementary amplicon sequencing confirmed the microbial community composition characteristic of high microbial abundance (HMA) and low microbial abundance (LMA) sponges. By use of GeoChip, altogether 20 273 probes encoding for 627 functional genes and representing 16 gene categories were identified. Minimum curvilinear embedding analyses revealed a clear separation between the samples. The HMA/LMA dichotomy was stronger than any possible geographic pattern, which is shown here for the first time on the level of functional genes. However, upon inspection of individual genes, very few specific differences were discernible. Differences were related to microbial ammonia oxidation, ammonification, and archaeal autotrophic carbon fixation (higher gene abundance in sponges over seawater) as well as denitrification and radiation-stress-related genes (lower gene abundance in sponges over seawater). Except for few documented specific differences the functional gene repertoire between the different sources appeared largely similar. This study expands previous reports in that functional gene convergence is not only reported between HMA and LMA sponges but also between sponges and seawater.

Description: Following their transmission from the human to the mosquito with the bloodmeal, malaria parasites have to persevere in the mosquito midgut for approximately 1 d. During this period the parasites are highly vulnerable to factors of the mosquito midgut, including bacteria. We here aimed at determining the microbial diversity of gut bacteria of the Asian malaria vector Anophebs stephensi (Liston) during development and under different feeding regimes, including feeds on malaria parasite-infected blood. 16S rRNA and denaturing gradient gel electrophoresis analyses demonstrated an increasing reduction in the microbial diversity during mosquito development from egg to adult and identified the gram-negative bacterium Elizabethkingia meningoseptica King as the dominant species in the midgut of lab-reared male and female mosquitoes. E. meningoseptica is transmitted between generations and its predominance in the mosquito midgut was not altered by diet, when the gut microbiota was compared between sugar-fed and blood-fed female mosquitoes. Furthermore, feeds on blood infected with malaria parasites did not impact the presence of E. meningoseptica in the gut. Extracts from cultured E. meningoseptica were active against gram-positive and negative bacteria and yeast and against the blood and gametocyte transmission stages of the malaria parasite Plasmodium falciparum Welch. The antimicrobial and antiplasmodial activities of E. meningoseptica may account for its dominance in the midgut of the malaria vector.

Description: The aim of this study was to examine sponge orange band (SOB) disease affecting the prominent Caribbean sponge Xestospongia muta. Scanning and transmission electron microscopy revealed that SOB is accompanied by the massive destruction of the pinacoderm. Chlorophyll a content and the main secondary metabolites, tetrahydrofurans, characteristic of X. muta, were significantly lower in bleached than in healthy tissues. Denaturing gradient gel electrophoresis using cyanobacteria-specific 16S rRNA gene primers revealed a distinct shift from the Synechococcus/Prochlorococcus clade of sponge symbionts towards several clades of unspecific cyanobacteria, including lineages associated with coral disease (i.e. Leptolyngbya sp.). Underwater infection experiments were conducted by transplanting bleached cores into healthy individuals, but revealed no signs of SOB development. This study provided no evidence for the involvement of a specific microbial pathogen as an etiologic agent of disease; hence, the cause of SOB disease in X. muta remains unidentified.

Description: Ferredoxins are electron shuttles harbouring iron–sulfur clusters that connect multiple oxido-reductive pathways in organisms displaying different lifestyles. Some prokaryotes and algae express an isofunctional electron carrier, flavodoxin, which contains flavin mononucleotide as cofactor. Both proteins evolved in the anaerobic environment preceding the appearance of oxygenic photosynthesis. The advent of an oxygen-rich atmosphere proved detrimental to ferredoxin owing to iron limitation and oxidative damage to the iron–sulfur cluster, and many microorganisms induced flavodoxin expression to replace ferredoxin under stress conditions. Paradoxically, ferredoxin was maintained throughout the tree of life, whereas flavodoxin is absent from plants and animals. Of note is that flavodoxin expression in transgenic plants results in increased tolerance to multiple stresses and iron deficit, through mechanisms similar to those operating in microorganisms. Then, the question remains open as to why a trait that still confers plants such obvious adaptive benefits was not retained. We compare herein the properties of ferredoxin and flavodoxin, and their contrasting modes of expression in response to different environmental stimuli. Phylogenetic analyses suggest that the flavodoxin gene was already absent in the algal lineages immediately preceding land plants. Geographical distribution of phototrophs shows a bias against flavodoxin-containing organisms in iron-rich coastal/freshwater habitats. Based on these observations, we propose that plants evolved from freshwater macroalgae that already lacked flavodoxin because they thrived in an iron-rich habitat with no need to back up ferredoxin functions and therefore no selective pressure to keep the flavodoxin gene. Conversely, ferredoxin retention in the plant lineage is probably related to its higher efficiency as an electron carrier, compared with flavodoxin. Several lines of evidence supporting these contentions are presented and discussed.

Description: A PCR protocol for the detection of sulfur-oxidizing bacteria based on soxB genes that are essential for thiosulfate oxidation by sulfur-oxidizing bacteria of various phylogenetic groups which use the 'Paracoccus sulfur oxidation' pathway was developed. Five degenerate primers were used to specifically amplify fragments of soxB genes from different sulfur-oxidizing bacteria previously shown to oxidize thiosulfate. The PCR yielded a soxB fragment of approximately 1000 bp from most of the bacteria. Amino acid and nucleotide sequences of soxB from reference strains as well as from new isolates and environmental DNA from a hydrothermal vent habitat in the North Fiji Basin were compared and used to infer relationships of soxB between sulfur-oxidizing bacteria belonging to various 16S rDNA-based phylogenetic groups. Major phylogenetic lines derived from 16S rDNA were confirmed by soxB phylogeny. Thiosulfate-oxidizing green sulfur bacteria formed a coherent group by their soxB sequences. Likewise, clearly separated branches demonstrated the distant relationship of representatives of alpha-, beta-, and gamma-Proteobacteria including representative species of the former genus Thiobacillus (now Halothiobacillus - gamma-Proteobacteria, Thiobacillus - beta-Proteobacteria and Starkeya - alpha-Proteobacteria). This general picture emerged although apparent evidence for lateral transfer of the soxB gene is indicated and comparison of soxB phylogeny and 16S rDNA phylogeny points to the significance of this gene transfer in hydrothermal vent bacterial communities of the North Fiji Basin.

Description: The functional gene amoA was used to compare the diversity of ammonia-oxidizing bacteria (AOB) in the water column and sediment-water interface of the two freshwater lakes Plusssee and Schöhsee and the Baltic Sea. Nested amplifications were used to increase the sensitivity of amoA detection, and to amplify a 789-bp fragment from which clone libraries were prepared. The larger part of the sequences was only distantly related to any of the cultured AOB and is considered to represent new clusters of AOB within the Nitrosomonas/Nitrosospira group. Almost all sequences from the water column of the Baltic Sea and from 1-m depth of Schöhsee were related to different Nitrosospira clusters 0 and 2, respectively. The majority of sequences from Plusssee and Schöhsee were associated with sequences from Chesapeake Bay, from a previous study of Plusssee and from rice roots in Nitrosospira-like cluster A, which lacks sequences from Baltic Sea. Two groups of sequences from Baltic Sea sediment were related to clonal sequences from other brackish/marine habitats in the purely environmental Nitrosospira-like cluster B and the Nitrosomonas-like cluster. This confirms previous results from 16S rRNA gene libraries that indicated the existence of hitherto uncultivated AOB in lake and Baltic Sea samples, and showed a differential distribution of AOB along the water column and sediment of these environments

Description: Phototrophic bacteria are important primary producers of salt lakes in the Salar de Atacama and at times form visible mass developments within and on top of the lake sediments. The communities of phototrophic bacteria from two of these lakes were characterized by molecular genetic approaches using key genes for the biosynthesis of the photosynthetic apparatus in phototrophic purple bacteria (pufLM) and in green sulfur bacteria (fmoA). Terminal restriction fragment length polymorphism of the pufLM genes indicated high variability of the community composition between the two lakes and subsamples thereof. The communities were characterized by the dominance of a novel, so far undescribed lineage of pufLM containing bacteria and the presence of representatives related to known halophilic Chromatiaceae and Ectothiorhodospiraceae. In addition, the presence of BChl b-containing anoxygenic phototrophic bacteria and of aerobic anoxygenic bacteria was indicated. Green sulfur bacteria were not detected in the environmental samples, although a bacterium related to Prosthecochloris indicum was identified in an enrichment culture. This is the first comprehensive description of phototrophic bacterial communities in a salt lake of South America made possible only due to the application of the functional pufLM genes.

Description: The concentration of CO2 in global surface ocean waters is increasing due to rising atmospheric CO2 emissions, resulting in lower pH and a lower saturation state of carbonate ions. Such changes in seawater chemistry are expected to impact calcification in calcifying marine organisms. However, other physiological processes related to calcification might also be affected, including enzyme activity. In a mesocosm experiment, macroalgal communities were exposed to three CO2 concentrations (380, 665, and 1486 µatm) to determine how the activity of two enzymes related to inorganic carbon uptake and nutrient assimilation in Corallina officinalis, an abundant calcifying rhodophyte, will be affected by elevated CO2 concentrations. The activity of external carbonic anhydrase, an important enzyme functioning in macroalgal carbon-concentrating mechanisms, was inversely related to CO2 concentration after long-term exposure (12 weeks). Nitrate reductase, the enzyme responsible for reduction of nitrate to nitrite, was stimulated by CO2 and was highest in algae grown at 665 µatm CO2. Nitrate and phosphate uptake rates were inversely related to CO2, while ammonium uptake was unaffected, and the percentage of inorganic carbon in the algal skeleton decreased with increasing CO2. The results indicate that the processes of inorganic carbon and nutrient uptake and assimilation are affected by elevated CO2 due to changes in enzyme activity, which change the energy balance and physiological status of C. officinalis, therefore affecting its competitive interactions with other macroalgae. The ecological implications of the physiological changes in C. officinalis in response to elevated CO2 are discussed.

Description: The sheer scale of the metagenomic and metatranscriptomic datasets that are now available warrants the development of automated protocols for organizing, annotating and comparing the samples in terms of their metabolic profiles. We describe a user-friendly java program FROMP (Fragment Recruitment on Metabolic Pathways) for mapping and visualizing enzyme annotations onto the Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathways or custom-made pathways and comparing the samples in terms of their Pathway Completeness Scores,their relative Activity Scores or enzyme enrichment odds ratios. This program along with our fully-configurable PERL based annotation organization pipeline Meta2Pro (METAbolic PROfiling of META-omic data) offers a quick and accurate standalone solution for metabolic profiling of environmental samples or cultures from different treatments. Apart from pictorial comparisons, FROMP can also generate score matrices for multiple meta-omics samples which can be used directly by other statistical programs

Description: Effects of epibiotic bacteria associated with macroalgae on barnacle larval attachment were investigated. Eight bacterial isolates obtained from samples of three macroalga species were cultured as monospecies bacterial films and tested for their activity against barnacle (Amphibalanus improvisus) attachment in field experiments (Western Baltic Sea). Furthermore, natural biofilm communities associated with the surface of the local brown alga, Fucus vesiculosus, which were exposed to different temperatures (5, 15 and 20 °C), were harvested and subsequently tested. Generally, monospecies bacterial biofilms, as well as natural microbial assemblages, inhibited barnacle attachment by 20-67%. denaturing gradient gel electrophoresis fingerprints showed that temperature treatment shifted the bacterial community composition and weakened the repellent effects at 20 °C. Repellent effects were absent when settlement pressure of cyprids was high. Nonviable bacteria tended to repel cyprids when compared to the unfilmed surfaces. We conclude that biofilms can have a repellent effect benefiting the host by preventing heavy fouling on its surface. However, severe settlement pressure, as well as stressful temperature, may reduce the protective effects of the alga's biofilm. Our results add to the notion that the performance of F. vesiculosus may be reduced by multiple stressors in the course of global warming.

Description: Shallow coastal waters, where phototrophic purple sulfur bacteria (PSB) regularly form massive blooms, are subjected to massive diurnal and event-driven changes of physicochemical conditions including temperature and salinity. To analyze the ability of PSB to cope with these environmental factors and to compete in complex communities we have studied changes of the environmental community of PSB of a Baltic Sea lagoon under experimental enrichment conditions with controlled variation of temperature and NaCl concentration. For the first time, changes within a community of PSB were specifically analyzed using the photosynthetic reaction center genes pufL and M by RFLP and cloning experiments. The most abundant PSB phylotypes in the habitat were found along the NaCl gradient from freshwater conditions up to 7.5% NaCl. They were accompanied by smaller numbers of purple nonsulfur bacteria and aerobic anoxygenic phototrophic bacteria. Major components of the PSB community of the brackish lagoon were affiliated to PSB genera and species known as marine, halophilic or salt-tolerant, including species of Marichromatium, Halochromatium, Thiorhodococcus, Allochromatium, Thiocapsa, Thiorhodovibrio, and Thiohalocapsa. A dramatic shift occurred at elevated temperatures of 41 and 44°C when Marichromatium gracile became most prominent which was not detected at lower temperatures.

Description: The taxonomy of two sympatric surf clams Donax marincovichi Coan, 1983 and Donax obesulus Reeve, 1854, inhabiting the coastal Humboldt Current Upwelling System is revisited. Because both species are exploited by artisanal fisheries, it is essential to verify that they are indeed distinct species that have to be managed separately. In this study, both taxa were sampled across their shared distributional area and specimens were indentified according to their respective morphological characteristics. Although width/height and height/length ratios revealed significant differences within sampling areas, the two morphotypes were frequently incongruent for taxonomically important morphometric parameters. In addition, they showed no significant mitochondrial genetic differentiation within or among populations and exhibited indistinguishable sperm ultrastructure. We conclude that the two morphotypes do not represent distinct species and should be included together under D. obesulus.

Description: Analyses of clone libraries from water and sediments of different sites from Salar de Huasco, a high-altitude athalassohaline wetland in the Chilean Altiplano, revealed the presence of five unique clusters of uncultured Archaea that have not been previously reported or specifically assigned. These sequences were distantly related (83–96% sequence identity) to a limited number of other clone sequences and revealed no identity to cultured Archaea. The abundance of Archaea and Bacteria was estimated using qPCR and community composition was examined through the construction of clone libraries of archaeal 16S rRNA gene. Archaea were found to be dominant over Bacteria in sediments from two saline sites (sites H4: 6.31 × 104 and site H6: 1.37 × 104 μS cm−1) and in one of the water samples (freshwater from site H0: 607 μS cm−1). Euryarchaeotal sequences were more abundant than crenarchaeotal sequences. Many of the clone sequences (52%) were similar to uncultured archaeal groups found in marine ecosystems having identity values between 99% and 97%. A major fraction of the sequences (40%) were members of Methanobacteria, while others were included in the Marine Benthic Groups B and D, the Miscellaneous Crenarchaeotic Group, the Terrestrial Miscellaneous Euryarchaeotal Group, Marine Group I and Halobacteria. The presence of uncultured archaeal groups in Salar de Huasco extends their known distribution in inland waters, providing new clues about their possible function in the environment.

Description: Life at deep-sea hydrothermal vents depends on chemolithoautotrophic microorganisms as primary producers mediating the transfer of energy from hydrothermal fluids to higher trophic levels. A comprehensive molecular survey was performed with microbial communities in a mussel patch at the Irina II site of the Logatchev hydrothermal field by combining the analysis of 16S rRNA gene sequences with studies of functional key genes involved in biochemical pathways of sulfur oxidation–reduction (soxB, aprA) and autotrophic carbon fixation (aclB, cbbM, cbbL). Most significantly, major groups of chemoautotrophic sulfur oxidizers in the diffuse fluids differed in their biosynthetic pathways of both carbon fixation and sulfur oxidation. One important component of the community, the Epsilonproteobacteria, has the potential to grow chemoautotrophically by means of the reductive tricarboxylic acid cycle and to gain energy through the oxidation of reduced sulfur compounds using the Sox pathway. The majority of soxB and all retrieved aclB gene sequences were assigned to this group. Another important group in this habitat, the Gammaproteobacteria, may use the adenosine 5'-phosphosulfate pathway and the Calvin–Benson–Bassham cycle, deduced from the presence of aprA and cbbM genes. Hence, two important groups of primary producers at the investigated site might use different pathways for sulfur oxidation and carbon fixation.

Description: The activity of the human armpit microbiota triggers the formation of body odor. We used differential 16S rRNA gene (rDNA)- and rRNA-based terminal-restriction fragment length polymorphism fingerprinting in combination with cloning and sequencing to identify active members of the human armpit microbiota. DNA and RNA were isolated from skin scrub samples taken from both armpits of 10 preconditioned, healthy males. The fingerprint profiles indicated pronounced similarities between the armpit microbiota in the right and the left axillae of an individual test person, but larger differences between the axilla microbiota of different individuals. Using 16S rDNA and rRNA sequence data, the majority of peaks in the armpit profiles were assigned to bacteria affiliated with well-known genera of skin bacteria. The relative abundances of all groups were similar among the rDNA and rRNA samples, suggesting that all groups of armpit bacteria were active. Surprisingly, the relative abundance of sequences affiliated with Peptoniphilus sp. was by far and with statistical significance the highest in the rRNA samples of the right armpits. Thus, bacteria affiliated with Peptoniphilus sp. might have been particularly active in the right axillae of the test persons, possibly owing to the handedness of the test persons, which might cause different environmental conditions in the right axillae.

Description: Bacterial communities associated with the brown alga Laminaria saccharina from the Baltic Sea and from the North Sea were investigated using denaturing gradient gel electrophoresis and 16S rRNA gene clone libraries. The rhizoid, cauloid, meristem and phyloid revealed different 16S rRNA gene denaturing gradient gel electrophoresis banding patterns indicating a specific association of bacterial communities with different parts of the alga. Associations with cauloid and meristem were more specific, while less specific associations were obtained from the old phyloid. In addition, seasonal and geographical differences in the associated communities were observed. Results from 16S rRNA gene libraries supported these findings. Bacterial phylotypes associated with the alga were affiliated with the Alphaproteobacteria (nine phylotypes), Gammaproteobacteria (nine phylotypes) and the Bacteroidetes group (four phylotypes). A number of bacteria associated with other algae and other marine macroorganisms were among the closest relatives of phylotypes associated with L. saccharina.

Description: Microbial isolate Z143-1 found to be associated with an unidentified tunicate was characterized due to its significant antimicrobial activity. Z143-1 is similar to Pseudovibrio ascidiaceicola and Pseudovibrio denitrificans in morphological, physiological and biochemical characteristics, except for its ability to ferment glucose and produce a characteristic red pigment. Fatty acid methyl ester analysis revealed a predominance of the fatty acid 18:1 ω7c at 80.55%, at levels slightly lower than the Pseudovibrio denitrificans type strain DN34T (87.7%). The mol% G+C of Z143-1 is 54.02, relatively higher than the Pseudovibrio denitrificans type strain DN34T and Pseudovibrio ascidiaceicola with mol% G+C of 51.7 and 51.4, respectively. However, phylogenetic analysis of the 16S rRNA gene sequence of Z143-1 showed 100% similarity with the Pseudovibrio denitrificans type strain DN34T. In this study, the bacterium Z143-1 is reported as a new strain of Pseudovibrio denitrificans. While there is no report of a secondary metabolite for Pseudovibrio denitrificans, Z143-1 produces the red pigment heptylprodigiosin, also known as 16-methyl-15-heptyl-prodiginine, which shows anti-Staphylococcus aureus activity.

Description: We investigated the gelatinous carnivore zooplankton community in the coastal waters of northeast Taiwan during the period 20072010. The community assemblage was composed of 45 species, of which only 14 appeared recurrent in Taiwanese waters. Although there was no clear seasonality, higher richness and abundances occurred in spring and autumn. Examination of potential physical drivers of coastal biomass accumulation did not show any link with water mass transport; instead, peak events were associated with typhoon disturbances, suggesting a potential resource pulse effect.

Description: This study aimed to examine interactive effects between ocean acidification and temperature on the photosynthetic and growth performance of Neosiphonia harveyi. N. harveyi was cultivated at 10 and 17.5 °C at present (~380 µatm), expected future (~800 µatm), and high (~1500 µatm) pCO2. Chlorophyll a fluorescence, net photosynthesis, and growth were measured. The state of the carbon-concentrating mechanism (CCM) was examined by pH-drift experiments (with algae cultivated at 10 °C only) using ethoxyzolamide, an inhibitor of external and internal carbonic anhydrases (exCA and intCA, respectively). Furthermore, the inhibitory effect of acetazolamide (an inhibitor of exCA) and Tris (an inhibitor of the acidification of the diffusive boundary layer) on net photosynthesis was measured at both temperatures. Temperature affected photosynthesis (in terms of photosynthetic efficiency, light saturation point, and net photosynthesis) and growth at present pCO2, but these effects decreased with increasing pCO2. The relevance of the CCM decreased at 10 °C. A pCO2 effect on the CCM could only be shown if intCA and exCA were inhibited. The experiments demonstrate for the first time interactions between ocean acidification and temperature on the performance of a non-calcifying macroalga and show that the effects of low temperature on photosynthesis can be alleviated by increasing pCO2. The findings indicate that the carbon acquisition mediated by exCA and acidification of the diffusive boundary layer decrease at low temperatures but are not affected by the cultivation level of pCO2, whereas the activity of intCA is affected by pCO2. Ecologically, the findings suggest that ocean acidification might affect the biogeographical distribution of N. harveyi.

Description: The vast majority of microbes inhabiting the subseafloor remain uncultivated and their energy sources unknown. Thus, a focus of ocean drilling expeditions over the past decade has been to characterize the distribution of microbes associated with specific metabolic reactions. An important question has been whether microbes involved in key microbial processes, such as sulfate reduction and methanogenesis, differ fundamentally from their counterparts in surface environments. To this end, functional genes of anaerobic methane cycling (mcrA), sulfate reduction (dsrAB), acetogenesis (fhs), and dehalorespiration (rdhA) have been examined. A compilation of existing functional gene data suggests that subseafloor microbes involved in anaerobic methane cycling, sulfate reduction, acetogenesis, and dehalorespiration are not fundamentally different from their counterparts in the surface world. Moreover, quantifications of mcrA and dsrAB suggest that, unless the majority of subseafloor microbes involved in methane cycling and sulfate reduction are too genetically divergent to be detected with conventional methods, these processes only support a small fraction (〈 1%) of total microbial biomass in the deep biosphere. Ecological explanations for the observed trends, target processes and methods for future investigations, and strategies for tackling the unresolved issue of microbial contamination in samples obtained by ocean drilling are discussed.

Description: Epibiotic biofilms have the potential to control major aspects of the biology and ecology of their hosts. Their composition and function may thus be essential for the health of the host. We tested the influence of salinity on the composition of epibacterial communities associated with the brown macroalga Fucus vesiculosus. Algal individuals were incubated at three salinities (5, 19, and 25) for 14days and nonliving reference substrata (stones) were included in the experiment. Subsequently, the composition of their surface-associated bacterial communities was analyzed by 454 pyrosequencing of 16S rRNA gene sequences. Redundancy analysis revealed that the composition of epiphytic and epilithic communities significantly differed and were both affected by salinity. We found that 5% of 2494 epiphytic operational taxonomic units at 97% sequence similarity were responsible for the observed shifts. Epibacterial -diversity was significantly lower at salinity 5 but did not differ between substrata. Our results indicate that salinity is an important factor in structuring alga-associated epibacterial communities with respect to composition and/or diversity. Whether direct or indirect mechanisms (via altered biotic interactions) may have been responsible for the observed shifts is discussed.

Description: Polynoidae contains ~900 species within 18 subfamilies, some of them restricted to the deep sea. Macellicephalinae is the most diverse among these deep-sea subfamilies. In the abyssal Equatorial Pacific Ocean, the biodiversity of benthic communities is at stake in the Clarion-Clipperton Fracture Zone (CCFZ) owing to increased industrial interest in polymetallic nodules. The records of polychaetes in this region are scarce. Data gathered during the JPI Oceans cruise SO239 made a significant contribution to fill this gap, with five different localities sampled between 4000 and 5000 m depth. Benthic samples collected using an epibenthic sledge or a remotely operated vehicle resulted in a large collection of polynoids. The aims of this study are as follows: (1) to describe new species of deep-sea polynoids using morphology and molecular data (COI, 16S and 18S); and (2) to evaluate the monophyly of Macellicephalinae. Based on molecular and morphological phylogenetic analyses, ten subfamilies are synonymized with Macellicephalinae in order to create a homogeneous clade determined by the absence of lateral antennae. Within this clade, the Anantennata clade was well supported, being determined by the absence of a median antenna. Furthermore, 17 new species and four new genera are described, highlighting the high diversity hidden in the deep. A taxonomic key for the 37 valid genera of the subfamily Macellicephalinae is provided.

Description: While we know much about the evolutionary patterns of endosymbiotic organelle origins, we know less about how the actual process unfolded within each system. This is partly due to the massive changes endosymbiosis appears to trigger, and partly because most organelles evolved in the distant past. The dinotoms are dinoflagellates with diatom endosymbionts, and they represent a relatively recent but nevertheless obligate endosymbiotic association. We have carried out deep sequencing of both the host and endosymbiont transcriptomes from two dinotoms, Durinskia baltica and Glenodinium foliaceum, to examine how the nucleocytosolic compartments have functionally integrated. This analysis showed little or no functional reduction in either the endosymbiont or host, and no evidence for genetic integration. Rather, host and endosymbiont seem to be bound to each other via metabolites, such as photosynthate exported from the endosymbiont to the host as indicated by the presence of plastidic phosphate translocators in the host transcriptome. The host is able to synthesize starch, using plant-specific starch synthases, as a way to store imported photosynthate.

Description: In spite of considerable insights into the microbial diversity of marine sponges, quantitative information on microbial abundances and community composition remains scarce. Here, we established qPCR assays for the specific quantification of four bacterial phyla of representative sponge symbionts as well as the kingdoms Eubacteria and Archaea. We could show that the 16S rRNA gene numbers of Archaea, Chloroflexi, and the candidate phylum Poribacteria were 4–6 orders of magnitude higher in high microbial abundance (HMA) than in low microbial abundance (LMA) sponges and that actinobacterial 16S rRNA gene numbers were 1–2 orders higher in HMA over LMA sponges, while those for Cyanobacteria were stable between HMA and LMA sponges. Fluorescence in situ hybridization of Aplysina aerophoba tissue sections confirmed the numerical dominance of Chloroflexi, which was followed by Poribacteria. Archaeal and actinobacterial cells were detected in much lower numbers. By use of fluorescence-activated cell sorting as a primer- and probe-independent approach, the dominance of Chloroflexi, Proteobacteria, and Poribacteria in A. aerophoba was confirmed. Our study provides new quantitative insights into the microbiology of sponges and contributes to a better understanding of the HMA/LMA dichotomy.

Description: Metabolites give us a window into the chemistry of microbes and are split into two subclasses: primary and secondary. Primary metabolites are required for life whereas secondary metabolites have historically been classified as those appearing after exponential growth and are not necessarily needed for survival. Many microbial species are estimated to produce hundreds of metabolites and can be affected by differing nutrients. Using various analytical techniques, metabolites can be directly detected in order to elucidate their biological significance. Currently, a single experiment can produce anywhere from megabytes to terabytes of data. This big data has motivated scientists to develop informatics tools to help target specific metabolites or sets of metabolites. Broadly, it is imperative to identify clear biological questions before embarking on a study of metabolites (metabolomics). For instance, studying the effect of a transposon insertion on phenazine biosynthesis in Pseudomonas is a very different from asking what molecules are present in a specific banana-derived strain of Pseudomonas. This review is meant to serve as a primer for a ‘choose your own adventure’ approach for microbiologists with limited mass spectrometry expertise, with a strong focus on liquid chromatography mass spectrometry based workflows developed or optimized within the past five years.

Description: Spliceosomal introns are a hallmark of eukaryotic genes that are hypothesized to play important roles in genome evolution but have poorly understood origins. Although most introns lack sequence homology to each other, new families of spliceosomal introns that are repeated hundreds of times in individual genomes have recently been discovered in a few organisms. The prevalence and conservation of these introner elements (IEs) or introner-like elements in other taxa, as well as their evolutionary relationships to regular spliceosomal introns, are still unknown. Here, we systematically investigate introns in the widespread marine green alga Micromonas and report new families of IEs, numerous intron presence-absence polymorphisms, and potential intron insertion hot-spots. The new families enabled identification of conserved IE secondary structure features and establishment of a novel general model for repetitive intron proliferation across genomes. Despite shared secondary structure, the IE families from each Micromonas lineage bear no obvious sequence similarity to those in the other lineages, suggesting that their appearance is intimately linked with the process of speciation. Two of the new IE families come from an Arctic culture (Micromonas Clade E2) isolated from a polar region where abundance of this alga is increasing due to climate induced changes. The same two families were detected in metagenomic data from Antarctica-a system where Micromonas has never before been reported. Strikingly high identity between the Arctic isolate and Antarctic coding sequences that flank the IEs suggests connectivity between populations in the two polar systems that we postulate occurs through deep-sea currents. Recovery of Clade E2 sequences in North Atlantic Deep Waters beneath the Gulf Stream supports this hypothesis. Our research illuminates the dynamic relationships between an unusual class of repetitive introns, genome evolution, speciation, and global distribution of this sentinel marine alga. © 2015 The Author.

Description: The prasinophyte order Mamiellales contains several widespread marine picophytoplankton (≤2 μm diameter) taxa, including Micromonas and Ostreococcus. Complete genome sequences are available for two Micromonas isolates, CCMP1545 and RCC299. We performed in silico analyses of nitrogen transporters and related assimilation genes in CCMP1545 and RCC299 and compared these with other green lineage organisms as well as Chromalveolata, fungi, bacteria, and archaea. Phylogenetic reconstructions of ammonium transporter (AMT) genes revealed divergent types contained within each Mamiellales genome. Some were affiliated with plant and green algal AMT1 genes and others with bacterial AMT2 genes. Land plant AMT2 genes were phylogenetically closer to archaeal transporters than to Mamiellales AMT2 genes. The Mamiellales represent the first green algal genomes to harbor AMT2 genes, which are not found in Chlorella and Chlamydomonas or the chromalveolate algae analyzed but are present in oomycetes. Fewer nitrate transporter (NRT) than AMT genes were identified in the Mamiellales. NRT1 was found in all but CCMP1545 and showed highest similarity to Mamiellales and proteobacterial NRTs. NRT2 genes formed a bootstrap-supported clade basal to other green lineage organisms. Several nitrogen-related genes were colocated, forming a nitrogen gene cluster. Overall, RCC299 showed the most divergent suite of nitrogen transporters within the various Mamiellales genomes, and we developed TaqMan quantitative polymerase chain reaction primer-probes targeting a subset of these, as well as housekeeping genes, in RCC299. All those investigated showed expression either under standard growth conditions or under nitrogen depletion. Like other recent publications, our findings show a higher degree of "mixed lineage gene affiliations" among eukaryotes than anticipated, and even the most phylogenetically anomalous versions appear to be functional. Nitrogen is often considered a regulating factor for phytoplankton populations. This study provides a springboard for exploring the use and functional diversification of inorganic nitrogen transporters and related genes in eukaryotic phytoplankton. © 2010 The Author.

Description: The basal position of the Mamiellales (Prasinophyceae) within the green lineage makes these unicellular organisms key to elucidating early stages in the evolution of chlorophyll a/b-binding light-harvesting complexes (LHCs). Here, we unveil the complete and unexpected diversity of Lhc proteins in Ostreococcus tauri, a member of the Mamiellales order, based on results from complete genome sequencing. Like Mantoniella squamata, O. tauri possesses a number of genes encoding an unusual prasinophyte-specific Lhc protein type herein designated "Lhcp". Biochemical characterization of the complexes revealed that these polypeptides, which bind chlorophylls a, b, and a chlorophyll c-like pigment (Mg-2,4-divinyl-phaeoporphyrin a 5 monomethyl ester) as well as a number of unusual carotenoids, are likely predominant. They are retrieved to some extent in both reaction center I (RCI)- and RCII-enriched fractions, suggesting a possible association to both photosystems. However, in sharp contrast to previous reports on LHCs of M. squamata, O. tauri also possesses other LHC subpopulations, including LHCI proteins (encoded by five distinct Lhca genes) and the minor LHCII polypeptides, CP26 and CP29. Using an antibody against plant Lhca2, we unambiguously show that LHCI proteins are present not only in O. tauri, in which they are likely associated to RCI, but also in other Mamiellales, including M. squamata. With the exception of Lhcp genes, all the identified Lhc genes are present in single copy only. Overall, the discovery of LHCI proteins in these prasinophytes, combined with the lack of the major LHCII polypeptides found in higher plants or other green algae, supports the hypothesis that the latter proteins appeared subsequent to LHCI proteins. The major LHC of prasinophytes might have arisen prior to the LHCII of other chlorophyll a/b-containing organisms, possibly by divergence of a LHCI gene precursor. However, the discovery in O. tauri of CP26-like proteins, phylogenetically placed at the base of the major LHCII protein clades, yields new insight to the origin of these antenna proteins, which have evolved separately in higher plants and green algae. Its diverse but numerically limited suite of Lhc genes renders O. tauri an exceptional model system for future research on the evolution and function of LHC components. © The Author 2005. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved.

Description: These data suggest that calcium may be lost during heavy sweating conditions (up to 20 mg calcium/hour) and that this loss should be considered in establishing recommended allowances for calcium. It was observed that 7 men consuming 441 mg of calcium a day in a study extending for 48 days, excreted 8.1, 11.6 and 20.2 mg/hour of calcium when living at 70, 85 and 100°F. This accounted for 21.8, 25.1 and 33.2% of the total calcium excreted. These observations are important since they show an additional calcium loss, which has not been reported in previous calcium balance studies in the literature. It is questionable whether an individual, consuming a low calcium diet, ever really attains calcium balance (equilibrium), under heavy sweating conditions. It was observed that (a) the calcium excreted in sweat, in men working at a moderate rate in extreme heat (100°F), was still fairly high after acclimatization, averaging 17 mg/hour after the first 4 days, and (b) that the daily total calcium in sweat increased as the sweat rate increased. Therefore it appears that the calcium requirements may be increased under these conditions. It was shown that even after acclimatization the urinary calcium did not decrease in compensation for the losses of calcium in sweat. It is recognized that changes in the urinary excretion of calcium in adjusting to different levels of dietary calcium and the various other metabolic factors, may require months to achieve.

Description: Dinoflagellates harboring diatom endosymbionts (termed “dinotoms”) have undergone a process often referred to as “tertiary endosymbiosis”—the uptake of algae containing secondary plastids and integration of those plastids into the new host. In contrast to other tertiary plastids, and most secondary plastids, the endosymbiont of dinotoms is distinctly less reduced, retaining a number of cellular features, such as their nucleus and mitochondria and others, in addition to their plastid. This has resulted in redundancy between host and endosymbiont, at least between some mitochondrial and cytosolic metabolism, where this has been investigated. The question of plastidial redundancy is particularly interesting as the fate of the host dinoflagellate plastid is unclear. The host cytosol possesses an eyespot that has been postulated to be a remnant of the ancestral peridinin plastid, but this has not been tested, nor has its possible retention of plastid functions. To investigate this possibility, we searched for plastid-associated pathways and functions in transcriptomic data sets from three dinotom species. We show that the dinoflagellate host has indeed retained genes for plastid-associated pathways and that these genes encode targeting peptides similar to those of other dinoflagellate plastid-targeted proteins. Moreover, we also identified one gene encoding an essential component of the dinoflagellate plastid protein import machinery, altogether suggesting the presence of a functioning plastid import system in the host, and by extension a relict plastid. The presence of the same plastid-associated pathways in the endosymbiont also extends the known functional redundancy in dinotoms, further confirming the unusual state of plastid integration in this group of dinoflagellates.

Description: Marine prey and predators will respond to future climate through physiological and behavioral adjustments. However, our understanding of how such direct effects may shift the outcome of predator–prey interactions is still limited. Here, we investigate the effects of ocean warming and acidification on foraging behavior and biomass of a common prey (shrimps, Palaemon spp.) tested in large mesocosms harboring natural resources and habitats. Acidification did not alter foraging behavior in prey. Under warming, however, prey showed riskier behavior by foraging more actively and for longer time periods, even in the presence of a live predator. No effects of longer-term exposure to climate stressors were detected on prey biomass. Our findings suggest that ocean warming may increase the availability of some prey to predators via a behavioral pathway (i.e., increased risk-taking by prey), likely by elevating metabolic demand of prey species.

Description: Pelagic aquatic environments differ from terrestrial environments in being three-dimensional and relatively homogeneous, rather than two-dimensional and heterogeneous. The present paper examines the causes and consequences of these differences in the context of their influence on the interactions of animals with environmental light. Particular emphasis is placed on light as a determinant of effective modes of crypsis in the two different habitats. The terrestrial world has selected for the expression of crypticity in the form of superficial color patterns. The heterogeneity of this habitat has resulted in evolutionary divergence of these superficial color patterns, often in very closely-related animals. In contrast, in the homogeneous pelagic aquatic habitats, evolutionary convergence on three main forms of crypsis is evident: (1) transparency; (2) reflection of most, if not all visible wavelengths; and, (3) ventral bioluminescence as counterillumination; thus, to be cryptic most animals in these habitats use one or a combination of these modalities to variously transmit, reflect or mimic environmental light. In the present paper, special attention is given to transparency as the most prevalent, yet least understood, of these mechanisms that are used in predator-prey interactions.

Description: The sexual cycle of the cuttlefish, Sepia officinalis, from the northern part of the Bay of Biscay was followed over several years (1988 to 1990 and 1992 to 1993). Sucessive maturity stages are reached at the same time regardless of site in the northern part of the Bay. In this area, the majority of cuttlefish reproduce during their second year of life (group II) whereas the remainder reproduce in their first year (group I). The first visible signs of sexual development concern the testis in males and the genital tract in females. Males mature earlier than females: the first spermatophores appear in July (group II) and October (group I) while mature eggs appear from December (group II) and March (group I). The breeding season lasts from about mid-March to late June (3.5 months).

Description: Statolith microstructure was studied in two abundant planktonic cranchiids, Cranchia scabra (56 specimens, 38-127mm mantle lengh, ML) and Liocranchia reinhardti (34 specimens, 99-205mm ML) sampled in epipelagic waters of the western part of the Gulf of Guinea (tropical Atlantic). Growth increments were revealed in ground statoliths of both species. It was possible to distinguish two growth zones in statolith microstructure by their colour in reflected light of the microscope: the translucent postnuclear zone and pale white opaque zone. Assuming that growth increments in statoliths were produced daily, ages of the largest immature C. scabra and L. reinhardti were 166 and 146 days, respectively. Both cranchiids are fast-growing squids with growth rates in length resembling those of juveniles of tropical ommastrephids and Thysanoteuthis rhombus. Liocranchia reinhardti grows faster: its growth rate in ML is approximately twice that of same-aged C. scabra. The life cycle of both cranchiids consists of two phases. During their epipelagic phase, C. scabra and L. reinhardti feed and grow rapidly from paralarvae to immature young in the epipelagic waters, attaining 120-130 and 170-200 mm ML by ages of 4-5 months, respectively. Then they change their life style to a deepwater phase.

Description: Parsimony analysis of 29 finned and finless octopod taxa considered 66 anatomical and morphological characters to discover synapomorphies that unite monophyletic groups. The resultant cladogram (177 equally parsimonious trees at 191 steps, CI 0.429) resolved all relationships except those among the 16 exemplars of the Octopodidae included and those among Tremoctopus, Ocythoe and Argonauta. Bootstrap values of over 90% support the monophyly of the finned and finless octopods, relationships among the finned octopods, the bolitaenids and the monophyly of Haliphron, Tremoctopus, Ocythoe and Argonauta; bootstrap values for other nodes range from 57 to 79%. Among finned octopods, specimens representing Grimpoteuthis are basal, as Voss (1988a) suggested. Specimens of Opisthoteuthis represent a distinct lineage, and are sister taxon, in this analysis, of Cirroteuthis (although specimens of Stauroteuthis could not be included). New definitions of the genera Opisthoteuthis and Grimpoteuthis are provided to reflect their separate evolutionary histories rather than their overt morphological similarity. Among finless octopods, bolitaenids are basal. The monophyletic Octopodidae is the sister taxon to the clade containing the sister taxa Vitreledonella and Amphitretus, and Haliphron, Tremoctopus, Ocythoe and Argonauta. The Ctenoglossa and Heteroglossa, families grouped by shared radular dentition, are diphyletic and paraphyletic, respectively. The cladistic relationships demonstrate that both the Vitrele-donellidae and Idioctopodidae are junior synonyms of the Amphitretidae; despite conspicuous morphological differences separating these taxa, they share a recent evolutionary history.

Description: Length-frequency data on squid (Loligo forbesi) collected during trawling surveys in Scottish waters from 1980 to 1994 were analysed to describe temporal and spatial patterns in abundance and to examine the prospects for using survey abundance to forecast fishery abundance. Loligo was patchily distributed in space and time. Distribution patterns in the North Sea in February appeared to be strongly related to bottom temperature (squid avoided waters 〈7°C) and, to a lesser extent, salinity (more squid in more saline water). For other areas and times, no temperature or salinity data were available, but there were trends for squid on the west coast to be more abundant in westerly areas and higher latitudes, and for squid at Rockall to be more abundant in shallow water. Inter-annual trends in abundance differed between the North Sea, west coast and Rockall, but average survey abundances for the North Sea and west coast tended to be positively correlated. For the North Sea and west coast, survey abundance was positively correlated with fishery abundance for the same month and area, and average abundance for the February North Sea survey was a reasonable predictor of commercial CPUE in the autumn of the same year (the peak of the fishery). Some of the observed trends were consistent with the existence of a stock-recruitment relationship but may indicate that abundance in a given calendar year is linked to climatic factors.

Description: The microstructure, morphology and ontogenetic development of statoliths and age and growth of 405 planktonic paralarvae and 117 juveniles belonging to 10 species of gonatid squids (Cephalopoda, Oegopsida) were studied in the region of the continental slope in the western part of the Bering Sea (57°00′–61°30′N, 163°00′E–179°20′W). The statolith microstructure of all species was characterized by the presence of a large droplet-shaped nucleus and bipartite postnuclear zone divided into two by the first stress check, except for Berryteuthis magister which had only one stress check and an undivided postnuclear zone. In Gonatus spp., completion of development of the postnuclear zone coincided with full development of the central hook on the tentacular club. Daily periodicity of statolith growth increments was validated by maintaining 13 paralarvae of the four most abundant species in captivity. All species might be subdivided into two groups based on statolith microstructure, i.e. species with a central position of the nucleus within the first statolith check (Gonatopsis spp., Egonatus tinro and B.magister) and species with the nucleus shifted to the inner side of the first statolith check (Gonatus spp.). Comparative analysis of statolith morphology showed that paralarval statoliths have species-specific characters that allowed the construction of keys to identify species of gonatid paralarvae based on their statoliths. Analysis of paralarval growth using statoliths revealed that these cold-water planktonic gonatid paralarvae have fast growth rates, attaining a mantle length of 7–10 mm at 15–20 days and 20–25 mm at 35–70 days.

Description: An investigation was carried out on the recently discovered ‘giant’ extra large (XL) form of the squid Sthenoteuthis oualaniensis from the Arabian Sea. The sample consisted of 2 males, which have not been previously described, and 13 females. Diet composition, parasite loading, sucker ring dentition, biolumi-nescence and sexual dimorphism were examined and compared to known parameters of the medium (M) form. Reproductive strategy, potential fecundity, egg size distribution in the ovary and oviducts were examined in mature XL females. Evidence of multiple spawning in the giant form was also investigated. Overall body shape, bioluminescent structures and coloration of the giant form were similar to the M form, though the XL form had a smaller fin angle than the M form. The mature female XL form has a dorsal mantle length about twice that of a mature female M form. Adult females of the XL form have a dorsal mantle length about twice that of adult males of the same form. Differences between males and females were found in arm sucker ring dentition and parasite load, suggesting a difference in diet. This could be linked to size differences between the sexes. A strong correlation between ovary mass and mantle length was found (r2 = 0.64). Poor correlation was found between mantle length and oviduct mass (r2 = 0.128) and potential fecundity (r2 = 0.07). Potential fecundity ranged between 2–5 million eggs and the holding capacity of the oviducts was approximately 300, 000 eggs. This combined with the presence of spermatangia and the presence of food in the stomach suggest that the XL form is a multiple spawner. S. oualaniensis appears to have a plastic phenotype and has adapted to the Arabian Sea conditions by evolving the capacity to grow to a giant size.

Description: Eight planktonic egg masses of the diamond-shaped Thysanoteuthis rhombus observed from 1995 to 2000 are described. Four were found in the western Mediterranean and the others were found off the Canary Islands. The egg masses from the Canary Islands are the first records for the eastern Atlantic. All were found near the surface at the end of summer and the beginning of autumn. The planktonic egg masses were dense, resilient oblong cylinders with rounded tips ranging from 80 to 130 cm in length and between 15 and 20 cm in diameter. Egg capsule dimensions ranged from 2.8 to 3.4 mm and total length of the newly hatched paralarvae was between 2.5 and 2.8 mm. Each egg mass contained an estimated 24 100–43 800 eggs. Some new characters that should help identification of the paralarvae, such as arm formulae, presence of an incipient keel-shaped membrane on some arms, and the type and chromatophore pattern are given. Ecological factors influencing the presence and distribution of these egg masses are discussed.

Description: The metabolic demands of a rare paper nautilus, Argonauta nouryi, in the eastern tropical Pacific (ETP) are evaluated. After adjusting for temperature and size, the rates of oxygen consumption and of aerobic and anaerobic metabolic potential (as evidenced by citrate synthase and octopine dehydrogenase activities, respectively) of A. nouryi were much higher than those in holopelagic octopods that exhibit float-and-wait predation strategies. In fact, the rates were similar to those found in small epipelagic squids and benthic octopods. The critical oxygen partial pressure was 4.9 kPa at 20°C, suggesting that the strong oxygen minimum layer found at intermediate depths in the ETP may constrain the vertical distribution of A. nouryi to the upper few metres of the water column. We also report the occurrence of a chain of shelled females at the surface, in which each animal was attached, as if on the benthos, to the next individual in the chain. Although it may constitute an effective strategy to increase the rates of mate encounter in the vast open ocean, there may be an important ecological trade-off for such behaviour, namely the increase in visibility at the surface with concomitant attraction of predators.

Description: Climate change influences the Baltic Sea ecosystem via its effects on oceanography and biogeochemistry. Sea surface temperature has been projected to increase by 2 to 4 °C until 2100 due to global warming; the changes will be more significant in the northern areas and less so in the south. The warming up will also diminish the annual sea ice cover by 57% to 71%, and ice season will be one to three months shorter than in the early 21st century, depending on latitude. A significant decrease in sea surface salinity has been projected because of an increase in rainfall and decrease of saline inflows into the Baltic Sea. The increasing surface flow has, in turn, been projected to increase leaching of nutrients from the soil to the watershed and eventually into the Baltic Sea. Also, acidification of the seawater and sea-level rise have been predicted. Increasing seawater temperature speeds up metabolic processes and increases growth rates of many secondary producers. Species associated with sea ice, from salt brine microbes to seals, will suffer. Due to the specific salinity tolerances, species’ geographical ranges may shift by tens or hundreds of kilometres with decreasing salinity. A decrease in pH will slow down calcification of bivalve shells, and higher temperatures also alleviate establishment of non-indigenous species originating from more southern sea areas. Many uncertainties still remain in predicting the couplings between atmosphere, oceanography and ecosystem. Especially projections of many oceanographic parameters, such as wind speeds and directions, the mean salinity level, and density stratification, are still ambiguous. Also, the effects of simultaneous changes in multiple environmental factors on species with variable preferences to temperature, salinity, and nutrient conditions are difficult to project. There is, however, enough evidence to claim that due to increasing runoff of nutrients from land and warming up of water, primary production and sedimentation of organic matter will increase; this will probably enhance anoxia and release of phosphorus from sediments. Such changes may keep the Baltic Sea in an eutrophicated state for a long time, unless strong measures to decrease nutrient runoff from land are taken. Changes in the pelagic and benthic communities are anticipated. Benthic communities will change from marine to relatively more euryhaline communities and will suffer from hypoxic events. The projected temperature increase and salinity decline will contribute to maintain the pelagic ecosystem of the Central Baltic and the Gulf of Finland in a state dominated by cyanobacteria, flagellates, small-sized zooplankton and sprat, instead of diatoms, large marine copepods, herring, and cod. Effects vary from area to area, however. In particular the Bothnian Sea, where hypoxia is less common and rivers carry a lot of dissolved organic carbon, primary production will probably not increase as much as in the other basins. The coupled oceanography-biogeochemistry ecosystem models have greatly advanced our understanding of the effects of climate change on marine ecosystems. Also, studies on climate associated “regime shifts” and cascading effects from top predators to plankton have been fundamental for understanding of the response of the Baltic Sea ecosystem to anthropogenic and climatic stress. In the future, modeling efforts should be focusing on coupling of biogeochemical processes and lower trophic levels to the top predators. Also, fine resolution species distribution models should be developed and combined with 3-D modelling, to describe how the species and communities are responding to climate-induced changes in environmental variables.

Description: Stomach contents of a resident community of bottlenose dolphins (Tursiops truncatus) from the Sarasota Bay area of the westcentral coast of Florida were studied to examine potential factors leading to patterns of habitat use. Composition and size of prey were analyzed and correlated with feeding behavior of individual dolphins of known histories. Examination of stomach contents of 16 stranded dolphins revealed a diet composed exclusively of fish (15 species), most of which were associated with seagrasses in varying degrees. Observational records for 21 years showed that feeding typically occurred in shallow (2–3 m) waters and in the vicinity of seagrasses in 23% of cases. Dolphins usually fed alone or in small groups and on non-obligate schooling prey. The main species of prey were soniferous, an indication that passive listening may be important in detection of prey. The close agreement between species of fishes represented in stomach contents and habitat of prey, as indicated by observations of feeding, suggests that analysis of stomach contents is a reasonable approach for studying prey and feeding patterns of dolphins. Meadows of seagrass are one of the habitats of importance to dolphins in the Sarasota Bay area, and their protection is important for conservation of these animals.

Description: Scaphitids are heteromorph ammonites exhibiting morphological trends counter to of those of the other main heteromorph ammonite groups. These trends include the shortening of the body chamber, lateral compression of the whorl, closure of the coil, and more regular, spiral coiling. Scaphitid-like morphologies may have appeared in other heteromorphs, but the Scaphitaceae are monophyletic. The most primitive scaphitids are known from the Albian, but an important radiation occurred in the Western Interior Seaway of North America from the Santonian to Maastrichtian, some of which spread to the Old World. The scaphitid morphology is consistent with improved swimming abilities, but scaphitids remained associated with the seafloor and are best considered to have been nektobenthos similar to modern nautiluses.

Description: A bioenergetic model for two narwhal (Monodon monoceros) sub-populations was developed to quantify daily gross energy requirements and estimate the biomass of Greenland halibut (Reinhardtius hippoglossoides) needed to sustain the sub-populations for their 5-month stay on wintering grounds in Baffin Bay. Whales in two separate wintering grounds were estimated to require 700 tonnes (s.e. 300) and 90 tonnes (s.e. 40) of Greenland halibut per day, assuming a diet of 50% Greenland halibut. Mean densities and length distributions of Greenland halibut inside and outside of the narwhal wintering grounds were correlated with predicted whale predation levels based on diving behavior. The difference in Greenland halibut biomass between an area with high predation and a comparable area without whales, approximately 19 000 tonnes, corresponded well with the predicted biomass removed by the narwhal sub-population on a diet of 50–75% Greenland halibut.

Description: King Penguins (Aptenodytes patagonicus) from breeding islands in the Indian Ocean (Crozet and Kerguelen Islands) and the Atlantic Ocean (South Georgia and Falkland Islands) were equipped with global location sensors to compare their foraging patterns during different times of the year. In summer, all birds investigated traveled toward the Antarctic Polar Front (APF), irrespective of whether they bred to the north (Crozet Islands, Falkland Islands), within (Kerguelen Islands) or to the south (South Georgia) of this hydrographic feature. Whereas most birds remained north of the APF and foraged in waters of the Antarctic Polar Frontal Zone, some penguins also traveled, or remained (South Georgia), south of the APF and foraged in Antarctic waters. It appeared that food resources in the vicinity of the APF were sufficiently predictable to warrant travel of several hundred km by King Penguins for foraging. Data collected on the winter distribution of King Penguins indicated at least two different foraging strategies. Birds from the oceanic Crozet Islands foraged beyond the APF in the Antarctic waters, whereas birds from the Falkland Islands relied also on the resources provided by the highly diverse and productive slope of the Patagonian Shelf. However, despite these differences, in both cases minimum distances of sometimes more than 10 000 km were covered. Further research on the foraging habitats of King Penguins over the entire breeding season and the temporal and spatial changes of oceanographic features is necessary to obtain a comprehensive picture on the variability in the foraging ranges of King Penguins.

Description: Mechanisms that affect thermal tolerance of ectothermic organisms have recently received much interest, mainly due to global warming and climate-change debates in both the public and in the scientific community. In physiological terms, thermal tolerance of several marine ectothermic taxa can be linked to oxygen availability, with capacity limitations in ventilatory and circulatory systems contributing to oxygen limitation at extreme temperatures. The present review briefly summarizes the processes that define thermal tolerance in a model cephalopod organism, the cuttlefish Sepia officinalis, with a focus on the contribution of the cephalopod oxygen-carrying blood pigment, hemocyanin. When acutely exposed to either extremely high or low temperatures, cuttlefish display a gradual transition to an anaerobic mode of energy production in key muscle tissues once critical temperatures (Tcrit) are reached. At high temperatures, stagnating metabolic rates and a developing hypoxemia can be correlated with a progressive failure of the circulatory system, well before Tcrit is reached. However, at low temperatures, declining metabolic rates cannot be related to ventilatory or circulatory failure. Rather, we propose a role for hemocyanin functional characteristics as a major limiting factor preventing proper tissue oxygenation. Using information on the oxygen binding characteristics of cephalopod hemocyanins, we argue that high oxygen affinities (= low P50 values), as found at low temperatures, allow efficient oxygen shuttling only at very low venous oxygen partial pressures. Low venous PO2s limit rates of oxygen diffusion into cells, thus eventually causing the observed transition to anaerobic metabolism. On the basis of existing blood physiological, molecular, and crystallographical data, the potential to resolve the role of hemocyanin isoforms in thermal adaptation by an integrated molecular physiological approach is discussed.

Description: In this article, the concepts and background of regional climate modeling of the future Baltic Sea are summarized and state-of-the-art projections, climate change impact studies, and challenges are discussed. The focus is on projected oceanographic changes in future climate. However, as these changes may have a significant impact on biogeochemical cycling, nutrient load scenario simulations in future climates are briefly discussed as well. The Baltic Sea is special compared to other coastal seas as it is a tideless, semi-enclosed sea with large freshwater and nutrient supply from a partly heavily populated catchment area and a long response time of about 30 years, and as it is, in the early 21st century, warming faster than any other coastal sea in the world. Hence, policymakers request the development of nutrient load abatement strategies in future climate. For this purpose, large ensembles of coupled climate–environmental scenario simulations based upon high-resolution circulation models were developed to estimate changes in water temperature, salinity, sea-ice cover, sea level, oxygen, nutrient, and phytoplankton concentrations, and water transparency, together with uncertainty ranges. Uncertainties in scenario simulations of the Baltic Sea are considerable. Sources of uncertainties are global and regional climate model biases, natural variability, and unknown greenhouse gas emission and nutrient load scenarios. Unknown early 21st-century and future bioavailable nutrient loads from land and atmosphere and the experimental setup of the dynamical downscaling technique are perhaps the largest sources of uncertainties for marine biogeochemistry projections. The high uncertainties might potentially be reducible through investments in new multi-model ensemble simulations that are built on better experimental setups, improved models, and more plausible nutrient loads. The development of community models for the Baltic Sea region with improved performance and common coordinated experiments of scenario simulations is recommended.

Description: Age and growth rates of the Mediterranean short-finned squid Illex coindetii (Ommastrephidae) were studied using statoliths from 704 specimens collected during trawl surveys within the Strait of Sicily in April and October–November 1995. In both samples, almost the whole ontogenetic spectrum was present, from juveniles to spent adults. Most of the squid captured in April had hatched between October and December 1994 (autumn–winter hatched group, AW), whereas most captured in October–November had hatched between May and July 1995 with a peak in June (spring–summer hatched group, SS). Age of the AW squid did not exceed 240 days (mainly 200–210 d), while that of the SS squid did not exceed 191 days (mainly 170–180 d). Growth in length and weight was best described by the logistic growth function. In both groups, daily growth rates (DGR) of males decreased at younger ages than in females, and sexual dimorphism in sizes (females larger than males) became evident in maturing and mature adults. The AW squids grew rather slowly during their juvenile and immature phases, which occurred in cool winter and spring seasons. They were characterized by a maximum DGR in mantle length (ML) and body weight (BW) at old ages, a prolonged period of maturation and old ages of mature specimens. AW squids attained large sizes and spawned at ages of 6–7 months in spring–summer. Their progeny (SS group) grew rapidly during the warm summer and autumn seasons and achieved their maximum DGR in ML and BW at younger ages than their AW predecessors. However, their younger age at maturation (5–6 months) reduced their somatic growth during the late ontogenetic stages, resulting in rather small sizes at maturity and early spawning. It is concluded that I. coindetii in the Strait of Sicily produce at least two alternating generations each year, which are adapted to live in different seasonal environmental conditions.

Description: The food habits of the beaked skate were studied utilising 274 individuals obtained from the incidental catches of the Argentine hake (Merluccius hubbsi) fishery. The most important prey were the Argentine hake, the southern cod (Patagonotothen ramsayi), the Argentine shortfin squid (Illex argentinus), the isopod (Serolis schythei), the “raneya” (Raneya brasiliensis , Pisces: Ophidiidae), and the Argentine anchovy (Engraulis anchoita). A total of 45 prey species was identified. No differences in the diet between sexes, but significant differences among size classes and between immature and mature individuals were found. Two size-related dietary shifts previously reported in this species, at around 35 cm and 85 cm total length were confirmed and related to changes in habitat utilisation. The first shift entails a major change from benthic prey (mostly crustaceans) to demersal-benthic prey (mostly fishes) and the second change from demersal-benthic to demersal-pelagic prey (increased consumption of Argentine hake and decreased consumption of southern cod). The second shift coincides with sexual maturation and may reflect a behavioural response to maturation.

Description: Although morphology of firm-bodied shallow-water octopuses resists preservation-linked deformation, preservation and handling of deep-sea specimens of Graneledone are hypothesized to deform specimens by accelerating fluid loss from mantle and arm tissues. Principal components analysis finds that seven of 39 North Pacific specimens of the genus are exceptionally narrow; the remaining specimens show considerable morphological variation, independent of time in preservation. Five exceptionally narrow specimens, including some paratypes of G. pacifica, were collected and preserved by the same trawling study; a single unrecorded factor in their common preservation history may have caused their deformation. The high morphological variation among the remaining specimens, seemingly unique among octopodids, may reflect factors as subtle as how the specimen was stored in the jar. Two types of change with time in preservation are documented. The mantle wall of formalin-fixed specimens thins unpredictably after storage for 30 months in 70% ethanol, consistent with deformation in preservation. Skin tubercles on the dorsal mantle, important taxonomic characters, become more prominent during the first decade in preservation. To maximize the information specimens with fluid-rich tissues convey and to identify the sources of deformation, the full preservation history and measurements made prior to dissection should be permanently recorded.

Description: Fishing can change the biological diversity of an ecosystem substantially, and determining the effects of fishing on marine biodiversity requires a variety of data, chief among them the proper identification of organisms. Because identification of all organisms in an ecosystem is not currently possible, target taxonomic groups (taxa) must be selected. The current status of taxonomic information varies greatly among taxa and among geographic areas within taxa. Problems include nomenclature, diagnoses, and determination of taxonomic relationships. We provide examples of a variety of these problems. We then propose a series of criteria for evaluating available taxonomic information in determining the potential reliability of species identification, including recency and comprehensiveness of revisionary studies, methods used for systematic and phylogenetic studies, adequacy of documentation, and evidence of peer review. When the goal is to explore biodiversity, these criteria must be used differently from when measuring and monitoring biodiversity.

Description: The systematics and distribution of the cirrate octopod genus Grimpoteuthis in the north-east Atlantic are reviewed. Three new species are described and Grimpoteuthis wuelkeri (Grimpe, 1920) is redescribed. A new generic diagnosis is proposed. Five species of Grimpoteuthis are recognized in the north-east Atlantic. The type species, G. umbellata (Fisher, 1883) is known only from the type specimen, which is in such poor condition that comparison with recently captured material was not possible. G. wuelkeri is a large, slope species, caught between 1600 m and 2200 m in the north-east and north-west Atlantic. Of the three new species, both G. boylei and G. challengeri are large abyssal species. G. boylei is found in the north-east Atlantic at the Porcupine Abyssal Plain (PAP) and the Madeira Abyssal Plain and may be found at abyssal depths throughout the north-east Atlantic. G. challengeri is known from the PAP, with a single specimen from the north-west Atlantic. G. discoveryi is a small, lower slope and abyssal species found in the north-east Atlantic. The Grimpoteuthis species can be separated based on shell form, presence of a radula and posterior salivary glands, arrangement of suckers and cirri and gill morphology. Two species, G. megaptera Verrill and G. plena Verrill, have been described from the north-west Atlantic, but the types are either lost (G. megaptera) or in poor condition (G. plena), hindering comparisons. Material examined from the north-west Atlantic included G. wuelkeri, G. challengeri and at least two other species.

Description: The distribution of paralarvae and small juvenile cephalopods sampled by a rectangular midwater trawl (opening area 8 m2) over the continental shelf off Cape Santa Marta Grande, southern Brazil (28°09′S–29°56′S) during spring 1989 is discussed. An intrusion of Brazil Current Tropical Water (22°C; 36.5) separates warm, less-saline water (22°C; 35.2) from cooler, more-saline water (15°C; 36.4). Prevailing northeasterly winds led to upwelling of South Atlantic Central Water over the shelf, promoting high Chl a concentrations. Three species constituted 99% of the 628 cephalopods collected: Illex argentinus (n = 540; 4–40 mm mantle length, ML), Argonauta nodosa (n = 46; 2–19 mm ML), and Loligo sanpaulensis (n = 42, 2–21 mm ML). Segregation of I. argentinus juveniles of similar size suggests school formation as small as 10 mm ML. The presence of mature males along with fertilized female A. nodosa indicates mating early in life. There was a consistent and direct link between high plankton production and high densities of juvenile cephalopods through a short and ecologically efficient food chain. The relationship between production, pycnocline intensity, and the density of paralarvae and juveniles revealed suitable conditions for survival and growth during the upwelling season.

Description: Recent molecular studies investigating higher-level phylogenetics of coleoid cephalopods (octopuses, squids and cuttlefishes) have produced conflicting results. A wide range of sequence alignment and analysis methods are used in cephalopod phylogenetic studies. The present study investigated the effect of commonly used alignment and analysis methods on higher-level cephalopod phylogenetics. Two sequence homology methods: (1) eye alignment, (2) implied alignment, and three analysis methods: (1) parsimony, (2) maximum likelihood, (3) Bayesian methodologies, were employed on the longest sequence dataset available for the coleoid cephalopods, comprising three mitochondrial and six nuclear loci. The data were also tested for base composition heterogeneity, which was detected in three genes and resolved using RY coding. The Octopoda, Argonautoidea, Oegopsida and Ommastrephidae are monophyletic in the phylogenies resulting from each of the alignment and analysis combinations. Furthermore, the Bathyteuthidae are the sister taxon of the Oegopsida in each case. However many relationships within the Coleoidea differed depending upon the alignment and analysis method used. This study demonstrates how differences in alignment and analysis methods commonly used in cephalopod phylogenetics can lead to different, but often highly supported, relationships.

Description: Osmotic adaption by halophilic and halotolerant bacteria is generally achieved by the accumulation or synthesis of several organic solutes. Accumulation by uptake from the medium is preferred over biosynthesis. The chemical nature of the major solute is important in determining the degree of osmotolerance of the organism. Glycine betaine accumulation confers a greater degree of osmotolerance than proline, which in turn confers more osmotolerance than glutamate accumulation. The occurrence and uptake of these solutes in a variety of eubacteria is reviewed.

Description: Over recent years, several PCR primers have been described to amplify genes encoding the structural subunits of ammonia monooxygenase (AMO) from ammonia-oxidizing bacteria (AOB). Most of them target amoA, while amoB and amoC have been neglected so far. This study compared the nucleotide sequence of 33 primers that have been used to amplify different regions of the amoCAB operon with alignments of all available sequences in public databases. The advantages and disadvantages of these primers are discussed based on the original description and the spectrum of matching sequences obtained. Additionally, new primers to amplify the almost complete amoCAB operon of AOB belonging to Betaproteobacteria (betaproteobacterial AOB), a primer pair for DGGE analysis of amoA and specific primers for gammaproteobacterial AOB, are also described. The specificity of these new primers was also evaluated using the databases of the sequences created during this study.

Description: Age composition and growth rates of the squid Loligo vulgaris (Lamark, 1797) were studied by examination of growth increments within statoliths of 419 specimens (mantle length, ML, ranging from 32 to 400mm). The squid were obtained by monthly sampling from the catches of commercial trawls off southern Portugal between March and September, 1993. The total number of growth increments in the mounted and ground statoliths was counted using a semi-automatic image analysis system. ML was significantly correlated with both the statolith length, TSL and the number of increments, NI. The female statolith was slightly larger than the male statolith for the same mantle size. Growth rates of individuals showed high variability with an average estimated at 34.6mm month-1 for males and 33.5mm month-1 for females. Growth in length between 70 and 280 days was best described by a power function for both sexes. The growth index of the statolith (TSL/NI) decreased with individual growth. The result may be related with the onset of sexual maturation. L. vulgaris hatched throughout the year with two distinct peaks, in spring which is the main breeding period, and in autumn. The life cycle of the L. vulgaris population on the south Portugese shelf was completed in one year.

Description: A study of individual and group growth rates based on a method of growth determination using the gladius was carried out in two populations of the purple-back flying squid (Sthenoteuthis oualaniesis) in the Arabian Sea. The duration of the life cycle in all populations and size groups examined was approximately one year. The growth pattern of linear increase was either slightly S-shaped or almost linear, while growth in weight followed a power-type pattern. Generally, females grew faster than males. Use of the gladius as a recording structure makes it possible to study the real growth of a specimen, rather than the statistical correlation of length/weight versus age, as in the commonly used statolith method. Analysis of individual growth rates in large-sized sawning females revealed growth rhythm with roughly a one-month periodicity in which phases of rapid growth (17-21 days; 1.6-3,6mm per day) alternate with phases of slow growth (12-14 days; 0.4-1.2mm per day). This new method is simple to use and may be applied even on board a vessel during research cruises.

Description: Trophodynamics of meso-zooplankton in the North Sea (NS) were assessed at a site in the southern NS, and at a shallow and a deep site in the central NS. Offshore and neritic species from different ecological niches, including Calanus spp., Temora spp. and Sagitta spp., were collected during seven cruises over 14 months from 2007 to 2008. Bulk stable isotope (SI) analysis, phospholipid-derived fatty acid (PLFA) compositions, and δ 13CPLFA data of meso-zooplankton and particulate organic matter (POM) were used to describe changes in zooplankton relative trophic positions (RTPs) and trophodynamics. The aim of the study was to test the hypothesis that the RTPs of zooplankton in the North Sea vary spatially and seasonally, in response to hydrographic variability, with the microbial food web playing an important role at times. Zooplankton RTPs tended to be higher during winter and lower during the phytoplankton bloom in spring. RTPs were highest for predators such as Sagitta sp. and Calanus helgolandicus and lowest for small copepods such as Pseudocalanus elongatus and zoea larvae (Brachyura). δ 15NPOM-based RTPs were only moderate surrogates for animals’ ecological niches, because of the plasticity in source materials from the herbivorous and the microbial loop food web. Common (16:0) and essential (eicosapentaenoic acid, EPA and docosahexaenoic acid, DHA) structural lipids showed relatively constant abundances. This could be explained by incorporation of PLFAs with δ 13C signatures which followed seasonal changes in bulk δ 13CPOM and PLFA δ 13CPOM signatures. This study highlighted the complementarity of three biogeochemical approaches for trophodynamic studies and substantiated conceptual views of size-based food web analysis, in which small individuals of large species may be functionally equivalent to large individuals of small species. Seasonal and spatial variability was also important in altering the relative importance of the herbivorous and microbial food webs.

Description: Ubiquitous SAR11 Alphaproteobacteria numerically dominate marine planktonic communities. Because they are excruciatingly difficult to cultivate, there is comparatively little known about their physiology and metabolic responses to long- and short-term environmental changes. As surface oceans take up anthropogenic, atmospheric CO2, the consequential process of ocean acidification could affect the global biogeochemical significance of SAR11. Shipping accidents or inadvertent release of chemicals from industrial plants can have strong short-term local effects on oceanic SAR11. This study investigated the effect of 2.5-fold acidification of seawater on the metabolism of SAR11 and other heterotrophic bacterioplankton along a natural temperature gradient crossing the North Atlantic Ocean, Norwegian and Greenland Seas. Uptake rates of the amino acid leucine by SAR11 cells as well as other bacterioplankton remained similar to controls despite an instant ∼50% increase in leucine bioavailability upon acidification. This high physiological resilience to acidification even without acclimation, suggests that open ocean dominant bacterioplankton are able to cope even with sudden and therefore more likely with long-term acidification effects.

Description: in a recent publication von Brandt (1) gives a survey of all determinations of the calcium content of Baltic water hitherto published. He records in all 39 analyses made during the last century, which give us an idea of the order of magnitude of the calcium concentration; they cannot, however, serve for comparative purposes as in many cases chlorine determinations on the same water samples are lacking. Neither have we any clue for judging the accuracy of these analyses, the latest of which date from 1884. Fifty years later, in 1935, Za rin s and O z o 1 ins (8) published an extensive investi­gation of the water in the Bay of Riga and in the Baltic off the Latvian coast, their most westerly station nearly coinciding with the Finnish station F81 (Lat. 57° 22'N., Long. 19°57'E.) above the central depression of the Baltic. Their material comprised about 70 calcium analyses on water from all depths. Finally v o n B r a n d t in the above-mentioned paper publishes nearly 300 analyses of surface water collected in 1935 and 1936 during several voyages from Pillau to Helsingfors and back, and along the German coast as far as Kiel and back. The present material comprises analyses of only 48 samples of surface and bottom water collected during the summer cruise, in July 1935, of the s.s. "Nautilus" from the Gulf of Finland, the Gulf of Bothnia, and the northern half of the Baltic proper. In spite of the smaller number of samples this material is more comprehensive than the two preceding investigations in so far as it covers a greater area of the sea. I t was originally meant as a survey of the calcium content in these parts of the Baltic, but the surprisingly simple relationships between calcium content and chlorinity which it revealed, give the results far more scope than was expected.

Description: We evaluated the effects of self-selected diets on calcium absorption and calculated retention in girls during pre-, early, and late puberty. Dietary calcium absorption was measured in 51 girls aged 4.9–16.7 y by using a dual-tracer stable-isotope technique. We found that calcium intake was similar among girls of all ages and all degrees of pubertal development and was below the recommended dietary allowance (1200 mg/d) for 21 of 25 girls aged ≥ 11 y. The early pubertal period was associated with a higher percent of dietary calcium absorption (34.4 ± 11.9%) than was the prepubertal (27.7 ± 8.2%) or late pubertal periods (25.9 ± 7.8%). Calculated calcium retention averaged 132 ± 83 mg/d in prepubertal girls, 161 ±88 mg/d in early pubertal girls, and 44 ± 91 mg/d in late pubertal girls. Peak calcium retention during early puberty was far below that previously reported after higher calcium intakes. We conclude that peak periods for calcium retention for girls are in the pre- and early pubertal periods. The current calcium intake of American girls during the pubertal growth period may not enable maximal mineral retention; therefore, increased calcium intakes should be considered.

Description: Intense bottom-ice algal blooms, often dominated by diatoms, are an important source of food for grazers, organic matter for export during sea ice melt, and dissolved organic carbon. Sea-ice diatoms have a number of adaptations, including accumulation of compatible solutes, that allows them to inhabit this highly variable environment characterized by extremes in temperature, salinity, and light. In addition to protecting them from extreme conditions, these compounds present a labile, nutrient-rich source of organic matter, and include precursors to climate active compounds (e.g., dimethyl sulfide [DMS]), which are likely regulated with environmental change. Here, intracellular concentrations of 45 metabolites were quantified in three sea-ice diatom species and were compared to two temperate diatom species, with a focus on compatible solutes and free amino acid pools. There was a large diversity of metabolite concentrations between diatoms with no clear pattern identifiable for sea-ice species. Concentrations of some compatible solutes (isethionic acid, homarine) approached 1 M in the sea-ice diatoms, Fragilariopsis cylindrus and Navicula cf. perminuta, but not in the larger sea-ice diatom, Nitzschia lecointei or in the temperate diatom species. The differential use of compatible solutes in sea-ice diatoms suggests different adaptive strategies and highlights which small organic compounds may be important in polar biogeochemical cycles.

Description: This review shows that the presence of seagrass microbial community is critical for the development of seagrasses; from seed germination, through to phytohormone production and enhanced nutrient availability, and defence against pathogens and saprophytes. The tight seagrass-bacterial relationship highlighted in this review supports the existence of a seagrass holobiont and adds to the growing evidence for the importance of marine eukaryotic microorganisms in sustaining vital ecosystems. Incorporating a micro-scale view on seagrass ecosystems substantially expands our understanding of ecosystem functioning and may have significant implications for future seagrass management and mitigation against human disturbance.

Description: Comparable values have been obtained for endogenous fecal calcium in cattle by an “isotope dilution” method and a “comparative balance” method. Both procedures involve the use of labeled calcium, but they are different in principle. The “isotope dilution” method offers advantages in being independent of the availability of the dietary calcium and of the uncertainties of balance determinations.