Localization of ion-regulatory epithelia in embryos and hatchlings of two cephalopods

The tissue distribution and ontogeny of Na+/K+-ATPase has been examined as an indicator for ion-regulatory epithelia in whole animal sections of embryos and hatchlings of two cephalopod species: the squid Loligo vulgaris and the cuttlefish Sepia officinalis. This is the first report of the immunohistochemical localization of cephalopod Na+/K+-ATPase with the polyclonal antibody alpha (H-300) raised against the human alpha1-subunit of Na+/K+-ATPase. Na+/K+-ATPase immunoreactivity was observed in several tissues (gills, pancreatic appendages, nerves), exclusively located in baso-lateral membranes lining blood sinuses. Furthermore, large single cells in the gill of adult L. vulgaris specimens closely resembled Na+/K+-ATPase-rich cells described in fish. Immunohistochemical observations indicated that the amount and distribution of Na+/K+-ATPase in late cuttlefish embryos was similar to that found in juvenile and adult stages. The ion-regulatory epithelia (e.g., gills, excretory organs) of the squid embryos and paralarvae exhibited less differentiation than adults. Na+/K+-ATPase activities for whole animals were higher in hatchlings of S. officinalis (157.0 ± 32.4 µmol/g FM/h) than in those of L. vulgaris (31.8 ± 3.3 µmol/g FM/h). S. officinalis gills and pancreatic appendages achieved activities of 94.8 ± 18.5 and 421.8 ± 102.3 µmol ATP/g FM/h, respectively. High concentrations of Na+/K+-ATPase in late cephalopod embryos might be important in coping with the challenging abiotic conditions (low pH, high pCO2) that these organisms encounter inside their eggs. Our results also suggest a higher sensitivity of squid vs. cuttlefish embryos to environmental acid-base disturbances.

Egg and early larval stages of Baltic cod, Gadus morhua duirng ocean acidification experiments, 2012

The accumulation of carbon dioxide in the atmosphere will lower the pH in ocean waters, a process termed ocean acidification (OA). Despite its potentially detrimental effects on calcifying organisms, experimental studies on the possible impacts on fish remain scarce. While adults will most likely remain relatively unaffected by changes in seawater pH, early life-history stages are potentially more sensitive, due to the lack of gills with specialized ion-regulatory mechanisms. We tested the effects of OA on growth and development of embryos and larvae of eastern Baltic cod, the commercially most important fish stock in the Baltic Sea. Cod were reared from newly fertilized eggs to early non-feeding larvae in 5 different experiments looking at a range of response variables to OA, as well as the combined effect of CO2 and temperature. No effect on hatching, survival, development, and otolith size was found at any stage in the development of Baltic cod. Field data show that in the Bornholm Basin, the main spawning site of eastern Baltic cod, in situ levels of pCO2are already at levels of 1,100 µatm with a pH of 7.2, mainly due to high eutrophication supporting microbial activity and permanent stratification with little water exchange. Our data show that the eggs and early larval stages of Baltic cod seem to be robust to even high levels of OA (3,200 µatm), indicating an adaptational response to CO2.

Effect of increased pCO2 level on early shell development in great scallop (Pecten maximus Lamarck) larvae

As a result of high anthropogenic CO2 emissions, the concentration of CO2 in the oceans has increased, causing a decrease in pH, known as ocean acidification (OA). Numerous studies have shown negative effects on marine invertebrates, and also that the early life stages are the most sensitive to OA. We studied the effects of OA on embryos and unfed larvae of the great scallop (Pecten maximus Lamarck), at pCO(2) levels of 469 (ambient), 807, 1164, and 1599 µatm until seven days after fertilization. To our knowledge, this is the first study on OA effects on larvae of this species. A drop in pCO(2) level the first 12 h was observed in the elevated pCO(2) groups due to a discontinuation in water flow to avoid escape of embryos. When the flow was restarted, pCO(2) level stabilized and was significantly different between all groups. OA affected both survival and shell growth negatively after seven days. Survival was reduced from 45% in the ambient group to 12% in the highest pCO(2) group. Shell length and height were reduced by 8 and 15 %, respectively, when pCO(2) increased from ambient to 1599 µatm. Development of normal hinges was negatively affected by elevated pCO(2) levels in both trochophore larvae after two days and veliger larvae after seven days. After seven days, deformities in the shell hinge were more connected to elevated pCO(2) levels than deformities in the shell edge. Embryos stained with calcein showed fluorescence in the newly formed shell area, indicating calcification of the shell at the early trochophore stage between one and two days after fertilization. Our results show that P. maximus embryos and early larvae may be negatively affected by elevated pCO(2) levels within the range of what is projected towards year 2250, although the initial drop in pCO(2) level may have overestimated the effect of the highest pCO(2) levels. Future work should focus on long-term effects on this species from hatching, throughout the larval stages, and further into the juvenile and adult stages.

Responses of the metabolism of the larvae of Pocillopora damicornis to ocean acidification and warming

Ocean acidification and warming are expected to threaten the persistence of tropical coral reef ecosystems. As coral reefs face multiple stressors, the distribution and abundance of corals will depend on the successful dispersal and settlement of coral larvae under changing environmental conditions. To explore this scenario, we used metabolic rate, at holobiont and molecular levels, as an index for assessing the physiological plasticity of Pocillopora damicornis larvae from this site to conditions of ocean acidity and warming. Larvae were incubated for 6 hours in seawater containing combinations of CO2 concentration (450 and 950 µatm) and temperature (28 and 30°C). Rates of larval oxygen consumption were higher at elevated temperatures. In contrast, high CO2 levels elicited depressed metabolic rates, especially for larvae released later in the spawning period. Rates of citrate synthase, a rate-limiting enzyme in aerobic metabolism, suggested a biochemical limit for increasing oxidative capacity in coral larvae in a warming, acidifying ocean. Biological responses were also compared between larvae released from adult colonies on the same day (cohorts). The metabolic physiology of Pocillopora damicornis larvae varied significantly by day of release. Additionally, we used environmental data collected on a reef in Moorea, French Polynesia to provide information about what adult corals and larvae may currently experience in the field. An autonomous pH sensor provided a continuous time series of pH on the natal fringing reef. In February/March, 2011, pH values averaged 8.075±0.023. Our results suggest that without adaptation or acclimatization, only a portion of naïve Pocillopora damicornis larvae may have suitable metabolic phenotypes for maintaining function and fitness in an end-of-the century ocean.

Growth, survival, gene expression, microbiota and tryptic activity during feeding of Scophthalmus maximus first feeding larvae with beta-glucan (MacroGard)

Reflecting the natural biology of mass spawning fish aquaculture production of fish larvae is often hampered by high and unpredictable mortality rates. The present study aimed to enhance larval performance and immunity via the oral administration of an immunomodulator, beta-glucan (MacroGard®) in turbot (Scophthalmus maximus). Rotifers (Brachionus plicatilis) were incubated with or without yeast beta-1,3/1,6-glucan in form of MacroGard® at a concentration of 0.5 g/L. Rotifers were fed to first feeding turbot larvae once a day. From day 13 dph onwards all tanks were additionally fed untreated Artemia sp. nauplii (1 nauplius ml/L). Daily mortality was monitored and larvae were sampled at 11 and 24 dph for expression of 30 genes, trypsin activity and size measurements. Along with the feeding of beta-glucan daily mortality was significantly reduced by ca. 15% and an alteration of the larval microbiota was observed. At 11 dph gene expression of trypsin and chymotrypsin was elevated in the MacroGard® fed fish, which resulted in heightened tryptic enzyme activity. No effect on genes encoding antioxidative proteins was observed, whilst the immune response was clearly modulated by beta-glucan. At 11 dph complement component c3 was elevated whilst cytokines, antimicrobial peptides, toll like receptor 3 and heat shock protein 70 were not affected. At the later time point (24 dph) an anti-inflammatory effect in form of a down-regulation of hsp 70, tnf-alpha and il-1beta was observed. We conclude that the administration of beta-glucan induced an immunomodulatory response and could be used as an effective measure to increase survival in rearing of turbot.

Abundance and biomass of mesozooplankton in the Levantine Basin during the Bilim 2 cruise in April 2008

The Sesame dataset contains mesozooplankton data collected during April 2008 in the Levantine Basin (between 33.20 and 36.50 N latitude and between 30.99 and 31.008 E longitude). Mesozooplankton samples were collected by using a WP-2 closing net with 200 µm mesh size during day hours (07:00-18:00). Samples were taken from 0-50, 50-100, 100-200 m layers at 5 stations in Levantine Basin The dataset includes samples analyzed for mesozooplankton species composition, abundance and total mesozooplankton biomass. Sampling volume was estimated by multiplying the mouth area with the wire length. Sampling biomass was measured by weighing filters and then determined by sampling volume. The samples were sieved sequentially through meshes of 500 and 200 micron to separate the mesozooplankton into size fractions. The entire sample (1/2) or an aliquot of the taxon-specific mesozooplankton abundance and the total abundance of the mesozooplankton were was analyzed under the binocular microscope. Minimum 500 individuals of mesozooplankton were identified and numerated at higher taxonomic level. Taxonomic identification was done at the METU- Institute of Marine Sciences by Alexandra Gubanova,Tuba Terbiyik using the relevant taxonomic literatures. Mesozooplankton abundance and biomass were estimated by Zahit Uysal and Yesim Ak.

Ichthyoplankton density and zooplankton biomass in the Benguela upwelling system during MSM17/3 in January and February 2011

Ichthyoplankton density (fish eggs and larvae) and bulk zooplankton biomass in January/February 2011 were determined for 38 stations in the northern Benguela upwelling system, based on oblique Multinet hauls during the FS Maria S. Merian MSM17/3 cruise. A HYDROBIOS Multinet, type Midi (0.25 m**2 mouth area) was equipped with five nets of 500 µm-mesh size, temperature and oxygen probes, and an inner and outer flow meter to monitor the net's trajectory (for volume filtered calculations) as well as net clogging. The Multinet was handled over the side, towed horizontally at 2 knots. Winch speed when fearing was 0.5 or 0.3 m/s, heaving velocity 0.2 - 0.3 m/s. The Multinet was towed obliquely at 38 stations sampling the upper 200 m of the water column, which were divided into five different depth strata after inspection of temperature and oxygen concentration depth profiles. Ichthyoplankton densities and zooplankton biomass were calculated for each depth stratum (=single net) from total abundance and the volume of water filtered [individuals per m**3 and g wet weight per m**3, respectively]. In addition, densities and biomass were integrated over the area for each station [individuals per m**2], as sum of calculations for each net: Sum ([individuals per m**3]*Delta (depth bot[m]-depth top [m]).

Ichthyoplankton density and zooplankton biomass in the Benguela upwelling system during MSM19/1b in October 2011

Ichthyoplankton density (fish eggs and larvae) and bulk zooplankton biomass in October 2011 were determined for 22 stations in the northern Benguela upwelling system, based on oblique Multinet hauls during the FS Maria S. Merian MSM19/1b cruise. A HYDROBIOS Multinet, type Midi (0.25 m**2 mouth area) was equipped with five nets of 500 µm-mesh size, temperature and oxygen probes, and an inner and outer flow meter to monitor the net's trajectory (for volume filtered calculations) as well as net clogging. The Multinet was handled over the side, towed horizontally at 2 knots. Winch speed when fearing was 0.5 or 0.3 m/s, heaving velocity 0.2 - 0.3 m/s. The Multinet was towed obliquely at 22 stations sampling the upper 200 m of the water column, which were divided into five different depth strata after inspection of temperature and oxygen concentration depth profiles. Ichthyoplankton densities and zooplankton biomass were calculated for each depth stratum (=single net) from total abundance and the volume of water filtered [individuals per m**3 and g wet weight per m**3, respectively]. In addition, densities and biomass were integrated over the area for each station [individuals per m**2], as sum of calculations for each net: Sum ([individuals per m**3]*Delta (depth bot[m]-depth top [m]).

Abundance of mesozooplankton collected in October and November 1991 at 20 stations in the South Aegean Sea during O91

The O91- Mesozooplankton dataset is based on samples collected in mid October-mid November 1991 at 20 stations in the South Aegean, the SE.Ionian Sea and in NW Levantine. Samples were collected at discrete layers (from the surface till 300m. These data are published. Sampling volume was estimated by multiplying the mouth area with the wire length. The entire sample (for deep layers) or aliquot of Taxon-specific mesozooplankton abundance (1/4) (for the upper layer) was analyzed under the binocular microscope. Copepod and cladoceran species were identified and enumerated; the other zooplankters were identified and enumerated at higher taxonomic level (commonly named as zooplankton groups). Taxonomic identification was done by I.Siokou-Frangou, E.Christou, and N.Fragopoulu, using the relevant taxonomic literature. The entire sample (for deep layers) or aliquot of Mesozooplankton total abundance (1/4) (for the upper layer) was analyzed under the binocular microscope. All zooplankters were enumerated.

Experimental ocean acidification alters the allocation of metabolic energy

Energy is required to maintain physiological homeostasis in response to environmental change. Although responses to environmental stressors frequently are assumed to involve high metabolic costs, the biochemical bases of actual energy demands are rarely quantified. We studied the impact of a near-future scenario of ocean acidification [800 µatm partial pressure of CO2 (pCO2)] during the development and growth of an important model organism in developmental and environmental biology, the sea urchin Strongylocentrotus purpuratus. Size, metabolic rate, biochemical content, and gene expression were not different in larvae growing under control and seawater acidification treatments. Measurements limited to those levels of biological analysis did not reveal the biochemical mechanisms of response to ocean acidification that occurred at the cellular level. In vivo rates of protein synthesis and ion transport increased 50% under acidification. Importantly, the in vivo physiological increases in ion transport were not predicted from total enzyme activity or gene expression. Under acidification, the increased rates of protein synthesis and ion transport that were sustained in growing larvae collectively accounted for the majority of available ATP (84%). In contrast, embryos and prefeeding and unfed larvae in control treatments allocated on average only 40% of ATP to these same two processes. Understanding the biochemical strategies for accommodating increases in metabolic energy demand and their biological limitations can serve as a quantitative basis for assessing sublethal effects of global change. Variation in the ability to allocate ATP differentially among essential functions may be a key basis of resilience to ocean acidification and other compounding environmental stressors.

Ichthyoplankton density and zooplankton biomass in the northern Benguela upwelling system during D356 September 2010

Ichthyoplankton density (fish eggs and larvae) and bulk zooplankton biomass in September 2010 were determined for 10 stations in the northern Benguela upwelling system, based on oblique Multinet hauls during the RRS Discovery D356 cruise. A HYDROBIOS Multinet, type Midi (0.25 m**2 mouth area) was equipped with five nets of 500 µm-mesh size, temperature and oxygen probes, and an inner and outer flow meter to monitor the net's trajectory (for volume filtered calculations) as well as net clogging. The Multinet was handled over the side, towed horizontally at 2 knots. Winch speed when fearing was 0.5 or 0.3 m/s, heaving velocity 0.2 - 0.3 m/s. The Multinet was towed obliquely at 10 stations sampling the upper 200 m of the water column, which were divided into five different depth strata after inspection of temperature and oxygen concentration depth profiles. Ichthyoplankton densities and zooplankton biomass were calculated for each depth stratum (=single net) from total abundance and the volume of water filtered [individuals per m**3 and g wet weight per m**3, respectively]. In addition, densities and biomass were integrated over the area for each station [individuals per m**2], as sum of calculations for each net: Sum ([individuals per m**3]*Delta(depth bot[m]-depth top [m]).

Ichthyoplankton density and zooplankton biomass in the Benguela upwelling system during AFR258 in December 2009

IIchthyoplankton density (fish eggs and larvae) and bulk zooplankton biomass in December 2009 were determined for 22 stations in the Benguela upwelling system, based on oblique Multinet hauls during the FRS Africana cruise AFR258. A HYDROBIOS Multinet, type Midi (0.25 m**2 mouth area) was equipped with five nets of 500 µm-mesh size, temperature and oxygen probes, and an inner and outer flow meter to monitor the net's trajectory (for volume filtered calculations) as well as net clogging. The Multinet was handled over the side, towed horizontally at 2 knots. Winch speed when fearing was 0.5 or 0.3 m/s, heaving velocity 0.2 - 0.3 m/s. The Multinet was towed obliquely at 22 stations sampling the upper 200 m of the water column, which were divided into five different depth strata after inspection of temperature and oxygen concentration depth profiles. Ichthyoplankton densities and zooplankton biomass were calculated for each depth stratum (=single net) from total abundance and the volume of water filtered [individuals per m**3 and g wet weight per m**3, respectively]. Densities and biomass were integrated over the area for each station [individuals per m**2], as sum of calculations for each net: Sum ([individuals per m**3]*Delta (depth bot[m]-depth top [m]).

Ichthyoplankton density and zooplankton biomass in the Benguela upwelling system during MSM07/3 in March 2008

Ichthyoplankton density (fish eggs and larvae) and bulk zooplankton biomass in March 2008 were determined for 32 stations in the northern Benguela upwelling system, based on oblique Multinet hauls during the FS Maria S. Merian MSM07/3 cruise. A HYDROBIOS Multinet, type Midi (0.25 m**2 mouth area) was equipped with five nets of 500 µm-mesh size, temperature and oxygen probes, and an inner and outer flow meter to monitor the net's trajectory (for volume filtered calculations) as well as net clogging. The Multinet was handled over the side, towed horizontally at 2 knots. Winch speed when fearing was 0.5 or 0.3 m/s, heaving velocity 0.2 - 0.3 m/s. The Multinet was towed obliquely at 32 stations sampling the upper 200 m of the water column, which were divided into five different depth strata after inspection of temperature and oxygen concentration depth profiles. Ichthyoplankton densities and zooplankton biomass were calculated for each depth stratum (=single net) from total abundance and the volume of water filtered [individuals per m**3 and g wet weight per m**3, respectively]. In addition, densities and biomass were integrated over the area for each station [individuals per m**2], as sum of calculations for each net: Sum ([individuals per m**3]*Delta (depth bot[m]-depth top [m]).

Ichthyoplankton density and zooplankton biomass in the Benguela upwelling system during MSM18/4 in July and August 2011

Ichthyoplankton density (fish eggs and larvae) and bulk zooplankton biomass in September 2010 were determined for 10 stations in the northern Benguela upwelling system, based on oblique Multinet hauls during the RRS Discovery D356 cruise. A HYDROBIOS Multinet, type Midi (0.25 m**2 mouth area) was equipped with five nets of 500 µm-mesh size, temperature and oxygen probes, and an inner and outer flow meter to monitor the net's trajectory (for volume filtered calculations) as well as net clogging. The Multinet was handled over the side, towed horizontally at 2 knots. Winch speed when fearing was 0.5 or 0.3 m/s, heaving velocity 0.2 - 0.3 m/s. The Multinet was towed obliquely at 10 stations sampling the upper 200 m of the water column, which were divided into five different depth strata after inspection of temperature and oxygen concentration depth profiles. Ichthyoplankton densities and zooplankton biomass were calculated for each depth stratum (=single net) from total abundance and the volume of water filtered [individuals per m**3 and g wet weight per m**3, respectively]. In addition, densities and biomass were integrated over the area for each station [individuals per m**2], as sum of calculations for each net: Sum ([individuals per m**3]*Delta(depth bot[m]-depth top [m]).