Temperature and photoperiod effects on sex determination in Leuresthes tenuis (fish)

In some gonochoristic species, sex is influenced not only by genotype at conception but also by the environment that offspring experience during early ontogeny (termed environmental sex determination or ESD). ESD is thought to be adaptive when seasonal variations in environmental conditions provide a sex-specific fitness advantage. In vertebrates, temperature is the most common determinant of sex, and seasonal variation in temperature serves as a temporal cue of environmental quality such as length of the growing season. Some environments, however, lack strong seasonal temperature fluctuations and other cues, particularly photoperiod, may provide a more reliable indicator of the environment offspring enter. We tested this hypothesis by rearing the offspring of the California grunion (Leuresthes tenuis, Ayres), which experiences low seasonal temperature variation in nature, under common garden conditions at three temperature and two photoperiod treatments. Our experiments revealed that both temperature and photoperiod significantly affected sex ratios in L. tenuis. More females were produced at cooler temperatures and longer day lengths, which is consistent with female biased sex ratios early in the breeding season, and likely adaptive through increased female size and fecundity. To our knowledge, this is the first documented case of photoperiod-dependent sex determination in a gonochoristic vertebrate.

(Appendix) Vertebral number of topsmelt, Atherinops affinis, adults and offspring from different latitudes along the North-American Pacific coast

Organisms that are distributed across spatial climate gradients often exhibit adaptive local variations in morphological and physiological traits, but to what extent such gradients shape evolutionary responses is still unclear. Given the strong natural contrast in latitudinal temperature gradients between the North-American Pacific and Atlantic coast, we asked how increases in vertebral number (VN, known as Jordan's Rule) with latitude would differ between Pacific (Atherinops affinis) and Atlantic Silversides (Menidia menidia), two ecologically equivalent and taxonomically similar fishes with similar latitudinal distributions. VN was determined from radiographs of wild-caught adults (genetic + environmental differences) and its genetic basis confirmed by rearing offspring in common garden experiments. Compared to published data on VN variation in M. menidia (a mean increase of 7.0 vertebrae from 32 to 46°N, VN slope = 0.42/lat), the latitudinal VN increase in Pacific Silversides was approximately half as strong (a mean increase of 3.3 vertebrae from 28 to 43°N, VN slope = 0.23/lat). This mimicked the strong Atlantic (1.11°C/lat) versus weak Pacific latitudinal gradient (0.40°C/lat) in median annual sea surface temperature (SST). Importantly, the relationship of VN to SST was not significantly different between the two species (average slope = -0.39 vertebrae/°C), thus suggesting a common thermal dependency of VN in silverside fishes. Our findings provide novel support for the hypothesis that temperature gradients are the ultimate cause of Jordan's Rule, even though its exact adaptive significance remains speculative. A second investigated trait, the mode of sex determination in Atlantic versus Pacific Silversides, revealed patterns that were inconsistent with our expectation: M. menidia displays temperature-dependent sex determination (TSD) at low latitudes, where growing seasons are long or unconstrained, but also a gradual shift to genetic sex determination (GSD) with increasing latitude due to more and more curtailed growing seasons. Sex ratios in A. affinis, on the other hand, were independent of latitude and rearing temperature (indicating GSD), even though growing seasons are thermally unconstrained across most of the geographical distribution of A. affinis. This suggests that additional factors (e.g., longevity) play an important role in shaping the mode of sex determination in silverside fishes.

Temperature dependent growth and inorganic ion composition of haemolymph of juvenile Sepia officinalis, 2006

B Body wet weight and mantle length of juvenile Sepia officinalis were monitored over a peroid of five weeks. The animals had hatched in our aquarium system in Bremerhaven, Germany at 16°C and were descendants of individuals collected in the Oosterschelde estuary, Netherlands. Animals were kept in natural sea water at 10 or 17°C and fed small live shrimp (Palaemonetes varians) ad libitum daily. At the end of the experiment some animals kept at 17°C were sacrificed using ethanol. Haemolymph was withdrawn from the head vein using syringe and needle. Haemolymph samples were stored at -20°C until Na+, Cl-, K+, Mg2+, Ca2+ and SO42- concentrations were determined using ion chromatography. Mean body weight more that tripled at 17°C during the investigation period, while growth was impared by exposue to 10°C. Haemolymph ion concentrations were similar to those in sea water, except for sulphate. The concentration of this ion in the haemolymph was more that ten times lower than in sea water.

Chlorophyll content and temperature-dependent net photosynthesis and respiration rate in the lichen Cetraria aculeata

We studied polar and temperate samples of the lichen Cetraria aculeata to investigate whether genetical differences between photobionts are correlated with physiological properties of the lichen holobiont. Net photosynthesis and dark respiration (DR) at different temperatures (from 0 to 30 °C) and photon flux densities (from 0 to 1,200 ?mol/m**2/s) were studied for four populations of Cetraria aculeata. Samples were collected from maritime Antarctica, Svalbard, Germany and Spain, representing different climatic situations. Sequencing of the photobiont showed that the investigated samples fall in the polar and temperate clade described in Fernández-Mendoza et al. (2011, doi:10.1111/j.1365-294X.2010.04993.x). Lichens with photobionts from these clades differ in their temperature optimum for photosynthesis, maximal net photosynthesis, maximal DR and chlorophyll content. Maximal net photosynthesis was much lower in Antarctica and Svalbard than in Germany and Spain. The difference was smaller when rates were expressed by chlorophyll content. The same is true for the temperature optima of polar (11 °C) and temperate (15 and 17 °C) lichens. Our results indicate that lichen mycobionts may adapt or acclimate to local environmental conditions either by selecting algae from regional pools or by regulating algal cell numbers (chlorophyll content) within the thallus.

Temperature-dependent phenology of Plutella xylostella (Lepidoptera: Plutellidae)

Temperature-dependent population growth of diamondback moth (DBM) Plutella xylostella (L.), a prolific insect pest of crucifer vegetables, was studied under six constant temperatures in the laboratory. The objective of the study was to predict the impacts of temperature changes on the population of DBM at high-resolution scales along altitudinal gradients and under climate change scenarios. Non-linear functions were fitted on the data for modeling the development, mortality, longevity and oviposition of the pest. The best-fitted functions for each life stage were compiled for estimating the life table parameters of the species by stochastic simulations. To quantify the impacts on the pest, three indices (establishment, generation and activity) were computed using the estimates of life table parameters and temperature data obtained at local scale (current scenario 2013) and downscaled climate change data (future scenario 2055) from the AFRICLIM database. To measure and represent the impacts of temperature change along the altitude on the pest; the indices were mapped along the altitudinal gradients of Kilimanjaro and Taita Hills, in Tanzania and Kenya, respectively. Potential impact of the changes between climate scenarios 2013 and 2055 was assessed. The data files included in this database were utilized for the above analysis to develop temperature dependent phenology of Plutella xylostella to assess current and future distribution along eastern African Afromontanes.

Temperature Dependent Effects of Elevated CO2 on Shell Composition and Mechanical Properties of Hydroides elegans: Insights from a Multiple Stressor Experiment

The majority of marine benthic invertebrates protect themselves from predators by producing calcareous tubes or shells that have remarkable mechanical strength. An elevation of CO2 or a decrease in pH in the environment can reduce intracellular pH at the site of calcification and thus interfere with animal's ability to accrete CaCO3. In nature, decreased pH in combination with stressors associated with climate change may result in the animal producing severely damaged and mechanically weak tubes. This study investigated how the interaction of environmental drivers affects production of calcareous tubes by the serpulid tubeworm, Hydroides elegans. In a factorial manipulative experiment, we analyzed the effects of pH (8.1 and 7.8), salinity (34 and 27), and temperature (23°C and 29°C) on the biomineral composition, ultrastructure and mechanical properties of the tubes. At an elevated temperature of 29°C, the tube calcite/aragonite ratio and Mg/Ca ratio were both increased, the Sr/Ca ratio was decreased, and the amorphous CaCO3 content was reduced. Notably, at elevated temperature with decreased pH and reduced salinity, the constructed tubes had a more compact ultrastructure with enhanced hardness and elasticity compared to decreased pH at ambient temperature. Thus, elevated temperature rescued the decreased pH-induced tube impairments. This indicates that tubeworms are likely to thrive in early subtropical summer climate. In the context of climate change, tubeworms could be resilient to the projected near-future decreased pH or salinity as long as surface seawater temperature rise at least by 4°C.

Haemolymph inorganic ion composition, physiology and behaviour dependent on temperature and ambient magnesium concentration in early life history stages of Paralomis granulosa (Decapoda, Anomura, Lithodidae)

Marine brachyuran and anomuran crustaceans are completely absent from the extremely cold (-1.8 °C) Antarctic continental shelf, but caridean shrimps are abundant. This has at least partly been attributed to low capacities for magnesium excretion in brachyuran and anomuran lithodid crabs ([Mg2+]HL = 20-50 mmol/L) compared to caridean shrimp species ([Mg2+]HL = 5-12 mmol/L). Magnesium has an anaesthetizing effect and reduces cold tolerance and activity of adult brachyuran crabs. We investigated whether the capacity for magnesium regulation is a factor that influences temperature-dependent activity of early ontogenetic stages of the Sub-Antarctic lithodid crab Paralomis granulosa. Ion composition (Na+, Mg2+, Ca2+, Cl-, [SO4]2-) was measured in haemolymph withdrawn from larval stages, the first and second juvenile instars (crabs I and II) and adult males and females. Magnesium excretion improved during ontogeny, but haemolymph sulphate concentration was lowest in the zoeal stages. Neither haemolymph magnesium concentrations nor Ca2+:Mg2+ ratios paralleled activity levels of the life stages. Long-term (3 week) cold exposure of crab I to 1 °C caused a significant rise of haemolymph sulphate concentration and a decrease in magnesium and calcium concentrations compared to control temperature (9 °C). Spontaneous swimming activity of the zoeal stages was determined at 1, 4 and 9 °C in natural sea water (NSW, [Mg2+] = 51 mmol/L) and in sea water enriched with magnesium (NSW + Mg2+, [Mg2+] = 97 mmol/L). It declined significantly with temperature but only insignificantly with increased magnesium concentration. Spontaneous velocities were low, reflecting the demersal life style of the zoeae. Heart rate, scaphognathite beat rate and forced swimming activity (maxilliped beat rate, zoea I) or antennule beat rate (crab I) were investigated in response to acute temperature change (9, 6, 3, 1, -1 °C) in NSW or NSW + Mg2+. High magnesium concentration reduced heart rates in both stages. The temperature-frequency curve of the maxilliped beat (maximum: 9.6 beats/s at 6.6 °C in NSW) of zoea I was depressed and shifted towards warmer temperatures by 2 °C in NSW + Mg2+, but antennule beat rate of crab I was not affected. Magnesium may therefore influence cold tolerance of highly active larvae, but it remains questionable whether the slow-moving lithodid crabs with demersal larvae would benefit from an enhanced magnesium excretion in nature.

Metabolic responses to temperature stress under elevated pCO2 in Crepidula fornicata

In the current context of environmental change, ocean acidification is predicted to affect the cellular processes, physiology and behaviour of all marine organisms, impacting survival, growth and reproduction. In relation to thermal tolerance limits, the effects of elevated pCO2 could be expected to be more pronounced at the upper limits of the thermal tolerance window. Our study focused on Crepidula fornicata, an invasive gastropod which colonized shallow waters around European coasts during the 20th century. We investigated the effects of 10 weeks' exposure to current (380 µatm) and elevated (550, 750, 1,000 µatm) pCO2 on this engineer species using an acute temperature increase (1 °C/12 h) as the test. Respiration rates were measured on both males (small individuals) and females (large individuals). Mortality increased suddenly from 34 °C, particularly in females. Respiration rate in C. fornicata increased linearly with temperature between 18 and 34 °C, but no differences were detected between the different pCO2 conditions either in the regressions between respiration rate and temperature or in Q10 values. In the same way, condition indices were similar in all the pCO2 treatments at the end of the experiment, but decreased from the beginning of the experiment. This species was highly resistant to acute exposure to high temperature regardless of pCO2 levels, even though food was limited during the experiment. Crepidula fornicata appears to have either developed resistance mechanisms or a strong phenotypic plasticity to deal with fluctuations of physicochemical parameters in its habitat. This suggests that invasive species may be more resistant to future environmental changes than its native competitors.

In-situ Iron isotope measurements and mineral compositions from ODP Hole 206-1256D and IODP Hole 335-U1256D

In-situ Fe isotope measurements have been carried out to estimate the impact of the hydrothermal metamorphic overprint on the Fe isotopic composition of Fe-Ti-oxides and Fe-sulfides of the different lithologies of the drilled rocks from IODP Hole 1256D (eastern equatorial Pacific; 15 Ma crust formed at the East Pacific Rise). Most igneous rocks normally have a very restricted range in their 56Fe/54Fe ratio. In contrast, Fe isotope compositions of hot fluids (> 300 °C) from mid-ocean-ridge spreading centers define a narrow range that is shifted to lower delta 56Fe values by 0.2 per mil - 0.5 per mil as compared to igneous rocks. Therefore, it is expected that mineral phases that contain large amounts of Fe are especially affected by the interaction with a fluid that fractionates Fe isotopes during exsolution/precipitation of those minerals. We have used a femtosecond UV-Laser ablation system to determine mineral 56Fe/54Fe ratios of selected samples with a precision of < 0.1 per mil (2 sigma level) at micrometer-scale. We have found significant variations of the delta 56Fe (IRMM-014) values in the minerals between different samples as well as within samples and mineral grains. The overall observed scale of delta 56Fe (magnetite) in 1256D rocks ranges from - 0.12 to + 0.64 per mil, and of delta 56Fe (ilmenite) from - 0.77 to + 0.01 per mil. Pyrite in the lowermost sheeted dike section is clearly distinguishable from the other investigated lithological units, having positive delta 56Fe values between + 0.29 and + 0.56 per mil, whereas pyrite in the other samples has generally negative delta 56Fe values from - 1.10 to - 0.59 permil. One key observation is that the temperature dependent inter-mineral fractionations of Fe isotopes between magnetite and ilmenite are systematically shifted towards higher values when compared to theoretically expected values, while synthesized, well equilibrated magnetite-ilmenite pairs are compatible with the theoretical predictions. Theoretical considerations including beta-factors of different aqueous Fe-chlorides and Rayleigh-type fractionations in the presence of a hydrous, chlorine-bearing fluid can explain this observation. The disagreement between observed and theoretical equilibrium fractionation, the fact that magnetite, in contrast to ilmenite shows a slight downhole trend in the delta 56Fe values, and the observation of small scale heterogeneities within single mineral grains imply that a general re-equilibration of the magnetite-ilmenite pairs is overprinted by kinetic fractionation effects, caused by the interaction of magnetite/ilmenite with hydrothermal fluids penetrating the upper oceanic crust during cooling, or incomplete re-equilibration at low temperatures. Furthermore, the observation of significant small-scale variations in the 56Fe/54Fe ratios of single minerals in this study highlights the importance of high spatial-resolution-analyses of stable isotope ratios for further investigations.

Grazing rate of microzooplankton measured experimentally in the North Atlantic during the Maria S. Merian cruise MSM26, spring 2013

The microzooplankton grazing dilution experiments were conducted at stations 126, 127, 131 and 133-137, following Landry & Hassett (1982). Seawater samples (whole seawater - WSW) were taken via Niskin bottles mounted on to a CTD Rosette out of the chlorophyll maximum at each station. Four different dilution levels were prepared with WSW and GF/F filtered seawater - 100% WSW, 75% WSW, 50% WSW and 25% WSW. The diluted WSW was filled in 2.4 L polycarbonate bottles (two replicates for every dilution level). Three subsamples (250 - 500 mL depending on in situ chlorophyll) of the 100% WSW were filtered on to GF/F filters (25 mm diameter) and chlorophyll was extracted in 5 mL 96% ethanol for 12-24 hours. Afterwards it was measured fluorometrically before and after the addition of HCl with a Turner fluorometer according to Jespersen and Christoffersen (1987) on board of the ship. In addition, one 250 mL subsample of the 100% WSW was fixed in 2% Lugol (final concentration), to determine the microzooplankton community when back at the Institute for Hydrobiology and Fisheries Science in Hamburg. Also, one 50 mL subsample of the 100% WSW was fixed in 1 mL glutaraldehyde, to quantify bacteria abundance. The 2.4 L bottles were put in black mesh-bags, which reduced incoming radiation to approximately 50% (to minimize chlorophyll bleaching). The bottles were incubated for 24 hours in a tank on deck with flow-through water, to maintain in situ temperature. An additional experiment was carried out to test the effect of temperature on microzooplankton grazing in darkness. Therefore, 100% WSW was incubated in the deck tank and in two temperature control rooms of 5 and 15°C in darkness (two bottles each). The same was done with bottles where copepods were added (five copepods of Calanus finmarchicus in each bottle; males and females were randomly picked and divided onto the bottles). In addition, two 100% WSW bottles with five copepods each were incubated at in situ temperature at 100% light level (without mesh-bags). All experiments were incubated for 24 hours and afterwards two subsamples of each bottle were filtered on to GF/F filters (25 mm diameter); 500 - 1000 mL depending on in situ chlorophyll. One 250 mL subsample of one of the two replicates of each dilution level and each additional experiment (temperature and temperature/copepods) was fixed in 5 mL lugol for microzooplankton determination. One 50 mL subsample of one of the two 100% WSW bottles as well as of one of the additional experiments without copepods was fixed in 1 mL glutaraldehyde for bacteria determination later on. Copepods were fixed in 4% formaldehyde for length measurements and sex determination.

Results from experiments and investigations on the kelp crab Taliepus dentatus

Studies of thermal tolerance in marine ectotherms are key in understanding climate effects on ecosystems; however, tolerance of their larval stages has rarely been analyzed. Larval stages are expected to be particularly sensitive. Thermal stress may affect their potential for dispersal and zoogeographical distribution. A mismatch between oxygen demand and the limited capacity of oxygen supply to tissues has been hypothesized to be the first mechanism restricting survival at thermal extremes. Therefore, thermal tolerance of stage zoea I larvae was examined in two populations of the Chilean kelp crab Taliepus dentatus, which are separated by latitude and the thermal regime. We measured temperature-dependent activity, oxygen consumption, cardiac performance, body mass and the carbon (C) and nitrogen (N) composition in order to: (1) examine thermal effects from organismal to cellular levels, and (2) compare the thermal tolerance of larvae from two environmental temperature regimes. We found that larval performance is affected at thermal extremes indicated by decreases in activity, mainly in maxilliped beat rates, followed by decreases in oxygen consumption rates. Cardiac stroke volume was almost temperature-independent. Through changes in heart rate, cardiac output supported oxygen demand within the thermal window whereas at low and high temperature extremes heart rate declined. The comparison between southern and central populations suggests the adaptation of southern larvae to a colder temperature regime, with higher cardiac outputs due to increased cardiac stroke volumes, larger body sizes but similar body composition as indicated by similar C:N ratios. This limited but clear differentiation of thermal windows between populations allows the species to widen its biogeographical range.

Marine meso-herbivore consumption scales faster with temperature than seaweed primary production, supplementary material

Respiration of ectotherms is predicted to increase faster with rising environmental temperature than photosynthesis of primary producers because of the differential temperature dependent kinetics of the key enzymes involved. Accordingly, if biological processes at higher levels of complexity are constrained by underlying metabolic functions food consumption by heterotrophs should increase more rapidly with rising temperature than photo-autoptrophic primary production. We compared rates of photosynthesis and growth of the benthic seaweed Fucus vesiculosus with respiration and consumption of the isopod Idotea baltica to achieve a mechanistic understanding why warming strengthens marine plant-herbivore interactions. In laboratory experiments thallus pieces of the seaweed and individuals of the grazer were exposed to constant temperatures at a range from 10 to 20°C. Photosynthesis of F. vesiculosus did not vary with temperature indicating efficient thermal acclimation whereas growth of the algae clearly increased with temperature. Respiration and food consumption of I. baltica also increased with temperature. Grazer consumption scaled about 2.5 times faster with temperature than seaweed production. The resulting mismatch between algal production and herbivore consumption may result in a net loss of algal tissue at elevated temperatures. Our study provides an explanation for faster decomposition of seaweeds at elevated temperatures despite the positive effects of high temperatures on algal growth.

Temperature and d18O measurements from different ODP Sites at the shelf and upper slope of the Peru Margin

The first experimentally determined temperature dependent oxygen-18 fractionation factor between dolomite and water at low temperatures [Vasconcelos et al. 1995 doi:10.1130/G20992.1] allows now the precise calculation of temperatures during early diagenetic dolomite precipitation. We use d18O values of early diagenetic dolomite beds sampled during ODP Legs 112 and 201 on the Peru continental margin (Sites 1227, 1228 and 1229) [Meister et al. 2007, doi:10.1111/j.1365-3091.2007.00870.x] to calculate paleo-porewater temperatures at the time of dolomite precipitation. We assumed unaltered seawater d18O values in the porewater, which is supported by d18O values of the modern porewater presented in this study. The dolomite layers in the Pleistocene part of the sedimentary columns showed oxygen isotope temperatures up to 5 °C lower than today. Since Sites 1228 and 1229 are located at 150 and 250 m below sealevel, respectively, their paleo-porewater temperatures would be influenced by considerably colder surface water during glacial sealevel lowstands. Thus, Pleistocene dolomite layers in the Peru Continental margin probably formed during glacial times. This finding is consistent with a model for dolomite precipitation in the Peru Margin recently discussed by Meister et al. [Meister et al. 2007, doi:10.1111/j.1365-3091.2007.00870.x], where dolomite forms episodically at the sulphate methane interface. It was shown that the sulphate methane interface migrates upwards and downwards within the sedimentary column, but dolomite layers may only form when the sulphate-methane interface stays at a fixed depth for a sufficient amount of time. We hypothesize that the sulphate-methane interface persists within TOC-rich interglacial sediments, while this zone is buried by TOC-poor sedimentation during glacial times. Thus, the presented oxygen isotope data provide additional information on the timing of early diagenetic dolomite formation and a possible link between episodicity in dolomite formation and sealevel variations. A similar link between early diagenesis and oceanography may also explain spacing of dolomite layers in a Milankovitch type pattern observed in the geological record, such as in the Miocene Monterey Formation.