Temperature and light requirements for growth of two diatom species (Bacillariophyceae) isolated from an Arctic macroalga

In the present study, two abundant epiphyticdiatom taxa were isolated from the assimilation hairs ofthe brown macroalga Chordaria flagelliformis collected inthe Arctic Kongsfjorden (Spitsbergen, Norway), establishedas unialgal cultures and their growth rates determinedunder controlled photon fluence rate andtemperature conditions. Using morphological (light andscanning electron microscopy) and SSU rRNA gene databoth isolates (ROS D99 and ROS D125) were identifiedas members of a FragilariaSynedropsis clade. Themolecular data of ROS D99 and ROS D125 were notidentical to any other published sequence. While ROSD99 has been identified as Fragilaria barbararum mainlydue to the SEM characteristics, ROS D125 could not bedefinitely identified although morphological data speakfor Fragilaria striatula. Both diatom species showedsimilar growth rates at all temperatures and photon fluencerates tested. They grew well between 0 and 15Cwithoptimum temperatures of 1214C, but did not survive20C. Therefore, compared to Antarctic diatoms bothtaxa from Kongsfjorden can be characterised as eurythermalorganisms. Increasing photon fluence rates between2 and 15 lmol m2 s1 were accompanied with analmost twofold increase in growth rates, but photon fluencerates >15 lmol m2 s1 did not further enhancegrowth pointing to low light requirements. From thesedata optimum, minimum and maximum photon fluencerates and temperatures for growth can be assessed indicatingthat both diatoms are well acclimated to the fluctuatingenvironmental conditions in the Arctic habitat

Chytridiomycosis of marine diatoms : the role of stress physiology and resistance in parasite-host recognition and accumulation of defense molecules

Little is known about the role of chemotaxis in the location and attachment of chytrid zoospores to potential diatom hosts. Hypothesizing that environmental stress parameters affect parasite-host recognition, four chytrid-diatom tandem cultures (Chytridium sp./Navicula sp., Rhizophydium type I/Nitzschia sp., Rhizophydium type IIa/Rhizosolenia sp., Rhizophydium type IIb/Chaetoceros sp.) were used to test the chemotaxis of chytrid zoospores and the presence of potential defense molecules in a non-contact-co-culturing approach. As potential triggers in the chemotaxis experiments, standards of eight carbohydrates, six amino acids, five fatty acids, and three compounds known as compatible solutes were used in individual and mixed solutions, respectively. In all tested cases, the whole-cell extracts of the light-stressed (continuous light exposure combined with 6 h UV radiation) hosts attracted the highest numbers of zoospores (86%), followed by the combined carbohydrate standard solution (76%), while all other compounds acted as weak triggers only. The results of the phytochemical screening, using biomass and supernatant extracts of susceptible and resistant host-diatom cultures, indicated in most of the tested extracts the presence of polyunsaturated fatty acids, phenols, and aldehydes, whereas the bioactivity screenings showed that the zoospores of the chytrid parasites were only significantly affected by the ethanolic supernatant extract of the resistant hosts

The effect of re-dissolution solvents and HPLC columns on the analysis of mycosporine-like amino acids in the eulittoral macroalgae and

International audienceMany macroalgal species that are regularly exposed to high solar radiation such as the eulittoral green alga and the red alga synthesize and accumulate high concentrations of mycosporine-like amino acids (MAAs) as UV-sunscreen compounds. These substances are typically extracted with a widely used standard protocol following quantification by various high performance liquid chromatography (HPLC) techniques. However, further preparation steps prior to HPLC analysis as well as different HPLC column types have not been systematically checked regarding separation quality and reproducibility. Therefore pure methanol, distilled water and HPLC eluent were evaluated as re-dissolution solvent for dried and extracts, which were subsequently analyzed on three reversed-phase C8 and C18 HPLC columns. The data indicate that distilled water and the HPLC eluent gave almost identical peak patterns and MAA contents on the C8 and C18 columns. In contrast, the application of the widely used methanol led to double peaks or even the loss of specific peaks as well as to a strong decline in total MAA amounts ranging from about 35% of the maximum in to 80% of the maximum in . Consequently, methanol should be avoided as re-dissolution solvent for the HPLC sample preparation. An improved protocol for the MAA analysis in macroalgae in combination with a reliable C18 column is suggested

Are Bacteria The Main Producers of Hydrolytic Enzymes in Aquatic Environment

Bacteria play an important role in the decomposition of organic matter, which is a key process in aquatic microbial food webs as well as its application in water bioremediation processes. DOM (Dissolved Organic Matter) is the major fraction of organic matter in most aquatic environments. Most of DOM is present as high molecular weight compounds that cannot be taken up by bacteria directly. Therefore, they must be hydrolysed enzymatically to be transported across microbial cell membranes. Molecular fluorosensors have been used as artificial substrates to study hydrolytic enzymes in situ. The present study wants to investigate the following hypothesis i.e. bacteria are the main producers of hydrolytic enzyme in aquatic systems. There were not any significant correlations between bacteria and the investigated hydrolases (esterase, peptidase, and β-glucosidase) in the meso- to eutrophic aquatic systems near Rostock city – North East Germany, although a wide range of bacterial abundances were covered. Enzyme saturations were not detected in all samples which may be explained by a high KM indicating a low affinity of enzymes (1) and or the contribution of many enzymes with different kinetics to the respective substrate degradation (2). There is also increasing evidence that bacteria may not be the sole or dominant source of esterases, peptidases and β-glucosidases in aquatic ecosystems. Invertebrate, fungi and other eukaryotes (diatoms, protozoa etc.) must be considered as possible and even important producers of hydrolytic enzymes. Several other factors may influence the correlations of bacteria to hydrolytic enzyme activities, i.e. variations in the species composition (1), a wide ranged variability of hydrolytic activities influenced directly by other substrates (low enzyme affinity), element availability (N and P), temperature and other abiotic factors (2), the existence and persistence of enzymes caused by other (passive) processes (e.g. cell lysis) (3)

Unraveling the Photoprotective Response of Lichenized and Free-Living Green Algae (Trebouxiophyceae, Chlorophyta) to Photochilling Stress

Lichens and free-living terrestrial algae are widespread across many habitats and develop successfully in ecosystems where a cold winter limits survival. With the goal of comparing photoprotective responses in free-living and lichenized algae, the physiological responses to chilling and photochilling conditions were studied in three lichens and their isolated algal photobionts together as well as in a fourth free-living algal species. We specifically addressed the following questions: (i) Are there general patterns of acclimation in green algae under chilling and photochilling stresses? (ii) Do free-living algae exhibit a similar pattern of responses as their lichenized counterparts? (iii) Are these responses influenced by the selection pressure of environmental conditions or by the phylogenetic position of each species? To answer these questions, photosynthetic fluorescence measurements as well as pigment and low molecular weight carbohydrate pool analyses were performed under controlled laboratory conditions. In general, photochemical efficiency in all free-living algae decreased with increasing duration of the stress, while the majority of lichens maintained an unchanged photochemical activity. Nevertheless, these patterns cannot be generalized because the alga Trebouxia arboricola and the lichen Ramalina pollinaria (associated with Trebouxia photobionts) both showed a similar decrease in photochemical efficiency. In contrast, in the couple Elliptochloris bilobata-Baeomyces rufus, only the algal partner exhibited a broad physiological performance under stress. This study also highlights the importance of the xanthophyll cycle in response to the studied lichens and algae to photochilling stress, while the accumulation of sugars was not related to cold acclimation, except in the alga E. bilobata. The differences in response patterns detected among species can be mainly explained by their geographic origin, although the phylogenetic position should also be considered, especially in some species.FM received a fellowship for PhD students (2012-2015) from the Government of the Basque Country as well as a mobility grant which allowed the collaboration between University of Basque Country (UPV/EHU) and University of Rostock. LG thanks the Deutsche Forschungsgemeinschaft for financial support (DFG GU 1278/1-1). The work was also supported by the project BFU 2010-15021 and CTM2014-53902-C2-2-P from the MINECO and the European Regional Development Fund ERDF (FEDER), and by the Basque Government (UPV/EHU-GV IT-624-13 and UPV/EHU-GV IT-1018-16)

Effects of prolonged darkness and temperature on the lipid metabolism in the benthic diatom Navicula perminuta from the Arctic Adventfjorden, Svalbard

The Arctic represents an extreme habitat for phototrophic algae due to long periods of darkness caused by the polar night (~4 months darkness). Benthic diatoms, which dominate microphytobenthic communities in shallow water regions, can survive this dark period, but the underlying physiological and biochemical mechanisms are not well understood. One of the potential mechanisms for long-term dark survival is the utilisation of stored energy products in combination with a reduced basic metabolism. In recent years, water temperatures in the Arctic increased due to an ongoing global warming. Higher temperatures could enhance the cellular energy requirements for the maintenance metabolism during darkness and, therefore, accelerate the consumption of lipid reserves. In this study, we investigated the macromolecular ratios and the lipid content and composition of Navicula cf. perminuta Grunow, an Arctic benthic diatom isolated from the microphytobenthos of Adventfjorden (Svalbard, Norway), over a dark period of 8 weeks at two different temperatures (0 and 7 °C). The results demonstrate that N. perminuta uses the stored lipid compound triacylglycerol (TAG) during prolonged dark periods, but also the pool of free fatty acids (FFA). Under the enhanced temperature of 7 °C, the lipid resources were used significantly faster than at 0 °C, which could consequently lead to a depletion of this energy reserves before the end of the polar night. On the other hand, the membrane building phospho- and glycolipids remained unchanged during the 8 weeks darkness, indicating still intact thylakoid membranes. These results explain the shorter survival times of polar diatoms with increasing water temperatures during prolonged dark periods

Seasonal variations of Fucus vesiculosus fertility under ocean acidification and warming in the western Baltic

Ocean warming and acidification may substantially affect the reproduction of keystone species such as Fucus vesiculosus (Phaeophyceae). In four consecutive benthic mesocosm experiments, we compared the reproductive biology and quantified the temporal development of Baltic Sea Fucus fertility under the single and combined impact of elevated seawater temperature and pCO2 (1100 ppm). In an additional experiment, we investigated the impact of temperature (0–25°C) on the maturation of North Sea F. vesiculosus receptacles. A marked seasonal reproductive cycle of F. vesiculosus became apparent in the course of 1 year. The first appearance of receptacles on vegetative apices and the further development of immature receptacles of F. vesiculosus in autumn were unaffected by warming or elevated pCO2. During winter, elevated pCO2 in both ambient and warmed temperatures increased the proportion of mature receptacles significantly. In spring, warming and, to a lesser extent, elevated pCO2 accelerated the maturation of receptacles and advanced the release of gametes by up to 2 weeks. Likewise, in the laboratory, maturation and gamete release were accelerated at 15–25°C relative to colder temperatures. In summary, elevated pCO2 and/or warming do not influence receptacle appearance in autumn, but do accelerate the maturation process during spring, resulting in earlier gamete release. Temperature and, to a much lesser extent, pCO2 affect the temporal development of Fucus fertility. Thus, rising temperatures will mainly shift or disturb the phenology of F. vesiculosus in spring and summer, which may alter and/or hamper its ecological functions in shallow coastal ecosystems of the Baltic Sea

Model simulation of seasonal growth of Fucus vesiculosus in its benthic community

Numerical models are a suitable tool to quantify impacts of predicted climate change on complex ecosystems but are rarely used to study effects on benthic macroalgal communities. Fucus vesiculosus L. is a habitat-forming macroalga in the Baltic Sea and alarming shifts from the perennial Fucus community to annual filamentous algae are reported. We developed a box model able to simulate the seasonal growth of the Baltic Fucus-grazer-epiphyte system. This required the implementation of two state variables for Fucus biomass in units of carbon (C) and nitrogen (N). Model equations describe relevant physiological and ecological processes, such as storage of C and N assimilates by Fucus, shading effects of epiphytes or grazing by herbivores on both Fucus and epiphytes, but with species-specific rates and preferences. Parametrizations of the model equations and required initial conditions were based on measured parameters and process rates in the near-natural Kiel Outdoor Benthocosm (KOB) experiments during the Biological Impacts of Ocean Acidification project. To validate the model, we compared simulation results with observations in the KOB experiment that lasted from April 2013 until March 2014 under ambient and climate-change scenarios, that is, increased atmospheric temperature and partial pressure of carbon dioxide. The model reproduced the magnitude and seasonal cycles of Fucus growth and other processes in the KOBs over 1 yr under different scenarios. Now having established the Fucus model, it will be possible to better highlight the actual threat of climate change to the Fucus community in the shallow nearshore waters of the Baltic Sea

Decreased Susceptibility of Staphylococcus aureus Small-Colony Variants toward Human Antimicrobial Peptides

Staphylococcus aureus is a frequent resident of human nose and skin in many individuals, but it is also able to cause a variety of serious infections including those of the skin and soft tissue. There is increasing evidence that particularly persistent, relapsing, and difficult-to-treat infections caused by S. aureus are associated with the formation of the small-colony variant (SCV) phenotype. The aim of this study was to investigate the hypothesis that (i) skin-derived antimicrobial peptides (AMPs) exhibit a reduced activity against SCVs and (ii) that switching into the SCV phenotype may endow S. aureus with a decreased susceptibility toward the killing activity of human stratum corneum. Here, we show that clinically derived S. aureus SCVs are less susceptible to the bactericidal activity of different human skin-derived AMPs as compared with their isogenic corresponding wild-type strains. Similarly, a S. aureus hemB mutant displaying the SCV phenotype was less susceptible to the antimicrobial activity of AMPs than its hemB-complemented mutant. These findings were accompanied by a higher resistance of SCVs to the killing activity of human stratum corneum. Switching into the SCV phenotype may help S. aureus to subvert cutaneous innate defense, thus contributing to the establishment and persistence of infection