Mitochondrial metabolism and respiration adjustments following temperature acclimation in Daphnia magna.

Seasonal temperature changes and local variations in the water column challenge lentic zooplankton in their habitat. At the cellular level, exposure to varying temperatures affects the mitochondrial functional properties of poikilothermic organisms. Metabolic enzymes that supply reduced substrates to the electron transport chain and elements of the oxidative phosphorylation system must therefore adjust their activity and flux rates to the altered temperature conditions. In the present study, Daphnia magna respiration was analyzed in response to acute and chronic changes in ambient temperature. Oxygen consumption as well as substrate and electron flux rates were measured at the animals' acclimation temperature and at two additional acute temperatures. High activity of citrate synthase (CS) in cold-acclimated animals (10 °C) may have resulted from mitochondrial quantitative adjustments. However, thermal sensitivity of the functional properties of mitochondrial enzymes was greater in warm-acclimated animals (30 °C). In whole animals, temperature-induced changes were partly compensated by acclimation, but these changes were promoted by acclimation in the case of the mitochondrial electron transport chain. Thus, respiration realised in whole animals was limited by the provision of reduced substrates in the tricarboxylic acid cycle rather than by restrictions of the respiratory chain complexes. This may minimize production of reactive oxygen species and resulting damage and reduce waste of substrates from the animals' energy reserves. Still, the integrated biomarker response indicated increased defense against oxidative stress at elevated temperatures.

European Lobster Larval Development and Fitness Under a Temperature Gradient and Ocean Acidification.

Climate change combined with anthropogenic stressors (e.g. overfishing, habitat destruction) may have particularly strong effects on threatened populations of coastal invertebrates. The collapse of the population of European lobster (Homarus gammarus) around Helgoland constitutes a good example and prompted a large-scale restocking program. The question arises if recruitment of remaining natural individuals and program-released specimens could be stunted by ongoing climate change. We examined the joint effect of ocean warming and acidification on survival, development, morphology, energy metabolism and enzymatic antioxidant activity of the larval stages of the European lobster. Larvae from four independent hatches were reared from stage I to III under a gradient of 10 seawater temperatures (13-24°C) combined with moderate (∼470 µatm) and elevated (∼1160 µatm) seawater pCO2 treatments. Those treatments correspond to the shared socio-economic pathways (SSP), SSP1-2.6 and SSP5-8.5 (i.e. the low and the very high greenhouse gas emissions respectively) projected for 2100 by the Intergovernmental Panel on Climate Change. Larvae under the elevated pCO2 treatment had not only lower survival rates, but also significantly smaller rostrum length. However, temperature was the main driver of energy demands with increased oxygen consumption rates and elemental C:N ratio towards warmer temperatures, with a reducing effect on development time. Using this large temperature gradient, we provide a more precise insight on the aerobic thermal window trade-offs of lobster larvae and whether exposure to the worst hypercapnia scenario may narrow it. This may have repercussions on the recruitment of the remaining natural and program-released specimens and thus, in the enhancement success of future lobster stocks.

Low-frequency noise pollution impairs burrowing activities of marine benthic invertebrates.

Sounds from human activities such as shipping and seismic surveys have been progressively invading natural soundscapes and pervading oceanic ambient sounds for decades. Benthic invertebrates are important ecosystem engineers that continually rework the sediment they live in. Here, we tested how low-frequency noise (LFN), a significant component of noise pollution, affects the sediment reworking activities of selected macrobenthic invertebrates. In a controlled laboratory setup, the effects of acute LFN exposure on the behavior of three abundant bioturbators on the North Atlantic coasts were explored for the first time by tracking their sediment reworking and bioirrigation activities in noisy and control environments via luminophore and sodium bromide (NaBr) tracers, respectively. The amphipod crustacean Corophium volutator was negatively affected by LFN, exhibiting lower bioturbation rates and shallower luminophore burial depths compared to controls. The effect of LFN on the polychaete Arenicola marina and the bivalve Limecola balthica remained inconclusive, although A. marina displayed greater variability in bioirrigation rates when exposed to LFN. Furthermore, a potential stress response was observed in L. balthica that could reduce bioturbation potential. Benthic macroinvertebrates may be in jeopardy along with the crucial ecosystem-maintaining services they provide. More research is urgently needed to understand, predict, and manage the impacts of anthropogenic noise pollution on marine fauna and their associated ecosystems.

A common temperature dependence of nutritional demands in ectotherms.

In light of ongoing climate change, it is increasingly important to know how nutritional requirements of ectotherms are affected by changing temperatures. Here, we analyse the wide thermal response of phosphorus (P) requirements via elemental gross growth efficiencies of Carbon (C) and P, and the Threshold Elemental Ratios in different aquatic invertebrate ectotherms: the freshwater model species Daphnia magna, the marine copepod Acartia tonsa, the marine heterotrophic dinoflagellate Oxyrrhis marina, and larvae of two populations of the marine crab Carcinus maenas. We show that they all share a non-linear cubic thermal response of nutrient requirements. Phosphorus requirements decrease from low to intermediate temperatures, increase at higher temperatures and decrease again when temperature is excessive. This common thermal response of nutrient requirements is of great importance if we aim to understand or even predict how ectotherm communities will react to global warming and nutrient-driven eutrophication.

An integrated multiple driver mesocosm experiment reveals the effect of global change on planktonic food web structure.

Global change puts coastal marine systems under pressure, affecting community structure and functioning. Here, we conducted a mesocosm experiment with an integrated multiple driver design to assess the impact of future global change scenarios on plankton, a key component of marine food webs. The experimental treatments were based on the RCP 6.0 and 8.5 scenarios developed by the IPCC, which were Extended (ERCP) to integrate the future predicted changing nutrient inputs into coastal waters. We show that simultaneous influence of warming, acidification, and increased N:P ratios alter plankton dynamics, favours smaller phytoplankton species, benefits microzooplankton, and impairs mesozooplankton. We observed that future environmental conditions may lead to the rise of Emiliania huxleyi and demise of Noctiluca scintillans, key species for coastal planktonic food webs. In this study, we identified a tipping point between ERCP 6.0 and ERCP 8.5 scenarios, beyond which alterations of food web structure and dynamics are substantial.

The thermal tolerance of a tropical population of blue crab (Callinectes sapidus) modulates aerobic metabolism during hypoxia.

The blue crab Callinectes sapidus is a widespread ectothermic species that supports large fisheries. Physiology of temperate and subtropical populations of blue crabs are well studied; however, a lack of information exists on tropical populations. Given the low locomotion capabilities of C. sapidus adult blue crabs, natural selection should favor traits that shape a particular thermal niche reflected through tolerance modulation to dissolved oxygen (DO). This study was designed to evaluate the thermal window and hypoxia sensitivity of the blue crab population in the southern Gulf of Mexico. The effect of acclimation temperatures from 20 °C to 34 °C on thermal preference (TP), critical thermal limits (CT), and thermal metabolic scope (TMS) was assessed in normoxia. Metabolic rate regulation over oxygen partial pressure (pO2) gradient was evaluated through oxygen consumption measurements at different degrees of acute hypoxia. Callinectes sapidus was observed tending to specialize towards higher temperatures, showing a mean TP from 26 °C to 33 °C. The lowest performance of aerobic pathways was observed at the coldest regimes and the highest at the warmest ones with mean TMS value being 35 % greater at 34 °C than 20 °C. Patterns for metabolic regulation were dependent on the interaction between environmental temperature and DO, in which the interval from 29 °C to 34 °C provoked a 50 % reduction in oxygen consumption when exposed to ∼20% air saturation levels. The results obtained showed that blue crabs distributed in the southern Gulf of Mexico could be close to their oxygen-temperature tolerance limits, which has important implications when climate change effects on species re-distribution is considered.

Noisy waters can influence young-of-year lobsters' substrate choice and their antipredatory responses.

Offshore human activities lead to increasing amounts of underwater noise in coastal and shelf environments, which may affect commercially-important benthic invertebrate groups like the re-stocked Helgoland European lobster (Homarus gammarus) in the German Bight (North Sea). It is crucial to understand the impact tonal low-frequency noises, like maritime transport and offshore energy operations, may have on substrate choice and lobsters' behavior to assess potential benefits or bottlenecks of new hard-substrate artificial offshore environments that become available. In this study, we investigated the full factorial effect of a tonal low-frequency noise and predator presence on young-of-year (YOY) European lobsters' in a diurnal and nocturnal experiment. Rocks and European oyster shells (Ostrea edulis) were offered as substrate to YOY lobsters for 3 h. Video recordings (n = 134) allowed the identification of lobsters' initial substrate choice, diel activity and key behaviors (peeking, shelter construction, exploration and hiding). To ensure independence, YOY lobsters in the intermolt stage were randomly selected and assigned to the experimental tanks and used only once. We provide the first evidence that stressors alone, and in combination, constrain YOY lobsters' initial substrate choice towards rocks. During nighttime, the joint effect of exposure to a constant low-frequency noise and predator presence decreased antipredator behavior (i.e., hiding) and increased exploration behavior. Noise may thus interfere with YOY lobsters' attention and decision-making processes. This outcome pinpoints that added tonal low-frequency noise in the environment have the potential to influence the behavior of early-life stages of European lobsters under predator pressure and highlights the importance of including key benthic invertebrates' community relationships in anthropogenic noise risk assessments. Among others, effects of noise must be taken into consideration in plans involving the multi-use of any offshore area for decapods' stock enhancement, aquaculture, and temporary no-take zones.

Hypoxia Tolerance of 10 Euphausiid Species in Relation to Vertical Temperature and Oxygen Gradients.

Oxygen Minimum Zones prevail in most of the world's oceans and are particularly extensive in Eastern Boundary Upwelling Ecosystems such as the Humboldt and the Benguela upwelling systems. In these regions, euphausiids are an important trophic link between primary producers and higher trophic levels. The species are known as pronounced diel vertical migrators, thus facing different levels of oxygen and temperature within a 24 h cycle. Declining oxygen levels may lead to vertically constrained habitats in euphausiids, which consequently will affect several trophic levels in the food web of the respective ecosystem. By using the regulation index (RI), the present study aimed at investigating the hypoxia tolerances of different euphausiid species from Atlantic, Pacific as well as from Polar regions. RI was calculated from 141 data sets and used to differentiate between respiration strategies using median and quartile (Q) values: low degree of oxyregulation (0.25 < RI median < 0.5); high degree of oxyregulation (0.5 < RI median < 1; Q1 > 0.25 or Q3 > 0.75); and metabolic suppression (RI median, Q1 and Q3 < 0). RI values of the polar (Euphausia superba, Thysanoessa inermis) and sub-tropical (Euphausia hanseni, Nyctiphanes capensis, and Nematoscelis megalops) species indicate a high degree of oxyregulation, whereas almost perfect oxyconformity (RI median ≈ 0; Q1 < 0 and Q3 > 0) was identified for the neritic temperate species Thysanoessa spinifera and the tropical species Euphausia lamelligera. RI values of Euphausia distinguenda and the Humboldt species Euphausia mucronata qualified these as metabolic suppressors. RI showed a significant impact of temperature on the respiration strategy of E. hanseni from oxyregulation to metabolic suppression. The species' estimated hypoxia tolerances and the degree of oxyconformity vs. oxyregulation were linked to diel vertical migration behavior and the temperature experienced during migration. The results highlight that the euphausiid species investigated have evolved various strategies to deal with different levels of oxygen, ranging from species showing a high degree of oxyconformity to strong oxyregulation. Neritic species may be more affected by hypoxia, as these are often short-distance-migrators and only adapted to a narrow range of environmental conditions.

Twenty years of the 'Preparation for Oxidative Stress' (POS) theory: Ecophysiological advantages and molecular strategies.

Freezing, dehydration, salinity variations, hypoxia or anoxia are some of the environmental constraints that many organisms must frequently endure. Organisms adapted to these stressors often reduce their metabolic rates to maximize their chances of survival. However, upon recovery of environmental conditions and basal metabolic rates, cells are affected by an oxidative burst that, if uncontrolled, leads to (oxidative) cell damage and eventually death. Thus, a number of adapted organisms are able to increase their antioxidant defenses during an environmental/functional hypoxic transgression; a strategy that was interpreted in the 1990s as a "preparation for oxidative stress" (POS). Since that time, POS mechanisms have been identified in at least 83 animal species representing different phyla including Cnidaria, Nematoda, Annelida, Tardigrada, Echinodermata, Arthropoda, Mollusca and Chordata. Coinciding with the 20th anniversary of the postulation of the POS hypothesis, we compiled this review where we analyze a selection of examples of species showing POS-mechanisms and review the most recent advances in understanding the underlying molecular mechanisms behind those strategies that allow animals to survive in harsh environments.

Maturation trade-offs in octopus females and their progeny: energy, digestion and defence indicators.

Sexual maturation and reproduction influence the status of a number of physiological processes and consequently the ecology and behaviour of cephalopods. Using Octopus mimus as a study model, the present work was focused in the changes in biochemical compound and activity that take place during gonadal maturation of females and its consequences in embryo and hatchlings characteristics. To do that, a total of 31 adult females of O. mimus were sampled to follow metabolites (ovaries and digestive gland) and digestive enzyme activities (alkaline and acidic proteases) during physiological and functional maturation. Levels of protein (Prot), triacylglyceride (TG), cholesterol (Chol), glucose (Glu), and glycogen (Gly) were evaluated. Groups of eggs coming from mature females were also sampled along development and after hatching (paralarvae of 1 and 3 days old) to track metabolites (Prot, TG, Glu, Gly, TG, Chol), digestive enzymes activity (Lipase, alkaline proteases, and acidic proteases), and antioxidant/detoxification defence indicators with embryos development. Based on the data obtained, we hypothesized that immature females store Chol in their ovaries, probably from the food they ingested, but switch to TG reserves at the beginning of the maturation processes. At the same time, results suggest that these processes were energetically supported by Glu, obtained probably from Gly breakdown by gluconeogenic pathways. Also, was observed that embryos metabolites and enzyme activities (digestive and antioxidant/detoxification enzymes) where maintained without significant changes and in a low activity during the whole organogenesis, meaning that organogenesis is relatively not energetically costly. In contrast, after organogenesis, a mobilization of nutrients and activation of the metabolic and digestive enzymes was observed, together with increments in consumption of yolk and Gly, and reduction in lipid peroxidation. Derived from our results, we also have the hypothesis that reactive oxygen species (ROS) were produced during the metabolic processes that occurs in ovarian maturation. Those ROS may be in part transferred to the egg provoking a ROS charge to the embryos. The elimination of ROS in embryos started when the activity of the heart and the absorption of the yolk around stages XIV and XV were evident. Altogether, these processes allowed the paralarvae to hatch with buffered levels of ROS and with the antioxidant defence mechanisms ready to support further ROS production derived from paralarvae higher life stage requirements (feeding and metabolic demands).

Comparison of Aerobic Scope for Metabolic Activity in Aquatic Ectotherms With Temperature Related Metabolic Stimulation: A Novel Approach for Aerobic Power Budget.

Considering that swim-flume or chasing methods fail in the estimation of maximum metabolic rate and in the estimation of Aerobic Scope (AS) of sedentary or sluggish aquatic ectotherms, we propose a novel conceptual approach in which high metabolic rates can be obtained through stimulation of organism metabolic activity using high and low non-lethal temperatures that induce high (HMR) and low metabolic rates (LMR), This method was defined as TIMR: Temperature Induced Metabolic Rate, designed to obtain an aerobic power budget based on temperature-induced metabolic scope which may mirror thermal metabolic scope (TMS = HMR-LMR). Prior to use, the researcher should know the critical thermal maximum (CT max) and minimum (CT min) of animals, and calculate temperature TIMR max (at temperatures -5-10% below CT max) and TIMR min (at temperatures +5-10% above CT min), or choose a high and low non-lethal temperature that provoke a higher and lower metabolic rate than observed in routine conditions. Two sets of experiments were carried out. The first compared swim-flume open respirometry and the TIMR protocol using Centropomus undecimalis (snook), an endurance swimmer, acclimated at different temperatures. Results showed that independent of the method used and of the magnitude of the metabolic response, a similar relationship between maximum metabolic budget and acclimation temperature was observed, demonstrating that the TIMR method allows the identification of TMS. The second evaluated the effect of acclimation temperature in snook, semi-sedentary yellow tail (Ocyurus chrysurus), and sedentary clownfish (Amphiprion ocellaris), using TIMR and the chasing method. Both methods produced similar maximum metabolic rates in snook and yellowtail fish, but strong differences became visible in clownfish. In clownfish, the TIMR method led to a significantly higher TMS than the chasing method indicating that chasing may not fully exploit the aerobic power budget in sedentary species. Thus, the TIMR method provides an alternative way to estimate the difference between high and low metabolic activity under different acclimation conditions that, although not equivalent to AS may allow the standardized estimation of TMS that is relevant for sedentary species where measurement of AS via maximal swimming is inappropriate.

Relationship between organochlorine pesticides and stress indicators in hawksbill sea turtle (Eretmochelys imbricata) nesting at Punta Xen (Campeche), Southern Gulf of Mexico.

Data on the impact of environmental pollution on the homeostasis of sea turtles remains scarce, particularly in the Southern Gulf of Mexico. As many municipalities along the coastline of the Yucatan Peninsula do not rely on a waste treatment plant, these organisms could be particularly vulnerable. We searched for relationships between the presence of organochlorine pesticides (OCP) and the level of several oxidative and pollutant stress indicators of the hawksbill sea turtle (Eretmochelys imbricata) during the 2010 nesting season at Punta Xen (Campeche, Mexico). Of the 30 sampled sea turtles, endosulfans, aldrin related (aldrin, endrin, dieldrin, endrin ketone, endrin aldehyde) and dichlorodiphenyldichloroethylene (DDT) families were detected in 17, 21 and 26, respectively. Significant correlation existed between the size of sea turtles with the concentration of methoxychlor, cholinesterase activity in plasma and heptachlors family, and catalase activity and hexachlorohexane family. Cholinesterase activity in washed erythrocytes and lipid peroxidation were positively correlated with glutathione reductase activity. Antioxidant enzyme actions seem adequate as no lipids damages were correlated with any OCPs. Future studies are necessary to evaluate the effect of OCPs on males of the area due to the significant detection of methoxychlor, which target endocrine functioning and increases its concentration with sea turtles size.

Histophagous ciliate Pseudocollinia brintoni and bacterial assemblage interaction with krill Nyctiphanes simplex. I. Transmission process.

Histophagous ciliates of the genus Pseudocollinia cause epizootic events that kill adult female krill (Euphausiacea), but their mode of transmission is unknown. We compared 16S rRNA sequences of bacterial strains isolated from stomachs of healthy krill Nyctiphanes simplex specimens with sequences of bacterial isolates and sequences of natural bacterial communities from the hemocoel of N. simplex specimens infected with P. brintoni to determine possible transmission pathways. All P. brintoni endoparasitic life stages and the transmission tomite stage (outside the host) were associated with bacterial assemblages. 16S rRNA sequences from isolated bacterial strains showed that Photobacterium spp. and Pseudoalteromonas spp. were dominant members of the bacterial assemblages during all life phases of P. brintoni and potential pathobionts. They were apparently unaffected by the krill's immune system or the histophagous activity of P. brintoni. However, other bacterial strains were found only in certain P. brintoni life phases, indicating that as the infection progressed, microhabitat conditions and microbial interactions may have become unfavorable for some strains of bacteria. Trophic infection is the most parsimonious explanation for how P. brintoni infects krill. We estimated N. simplex vulnerability to P. brintoni infection during more than three-fourths of their life span, infecting mostly adult females. The ciliates have relatively high prevalence levels (albeit at <10% of sampled stations) and a short life cycle (estimated <7 d). Histophagous ciliate-krill interactions may occur in other krill species, particularly those that form dense swarms and attain high population densities that potentially enhance trophic transmission and allow completion of the Pseudocollinia spp. life cycle.

Histophagous ciliate Pseudocollinia brintoni and bacterial assemblage interaction with krill Nyctiphanes simplex. II. Host responses.

Unlike decapod crustaceans of commercial interest, the krill defense system and its response to parasites and pathogens is virtually unknown. Histophagous ciliates of the genus Pseudocollinia interact with at least 7 krill species in the northeastern Pacific. Although they can cause epizootic events, the physiology of the histophagous ciliate-host interaction and krill (host) defenses remain unknown. From 1 oceanographic survey along the southwestern coast of the Baja California Peninsula near Bahía Magdalena and 2 in the Gulf of California, we investigated parasitoid-host physiological responses (fatty acid and oxidative stress indicators) of the subtropical krill Nyctiphanes simplex infected with the ciliate P. brintoni. All life stages of P. brintoni were associated with opportunistic bacterial assemblages that have not been explicitly investigated in other Pseudocollinia species (P. beringensis, P. oregonensis, and P. similis). Parasitoid ciliates exclusively infected adult females, which showed increased lipid content during gonad development. As the infection progressed, omega-3 eicosapentaenoic and docosahexaenoic fatty acids, which may act as energy sources to produce high numbers of ciliate transmission stages, were quickly depleted. Antioxidant enzymes, components of the crustacean defense system, varied throughout infection, but without inhibiting Pseudocollinia infection, i.e. higher levels of lipid oxidative damage were detected in late stages of infection. The ineffective response of the krill antioxidant defense system against histophagous ciliates and the bacteria associated with the ciliates suggests that Pseudocollinia ciliates are functionally analogous to krill predators and may have a strong influence on the population dynamics of krill.