Analysis of subcellular energy metabolism in five Lacertidae lizards across varied environmental conditions.

Aerobic respiration is the main energy source for most eukaryotes, and efficient mitochondrial energy transfer greatly influences organismal fitness. To survive environmental changes, cells have evolved to adjust their biochemistry. Thus, measuring energy metabolism at the subcellular level can enhance our understanding of individual performance, population dynamics, and species distribution ranges. We investigated three important metabolic traits at the subcellular level in five lacertid lizard species sampled from different elevations, from sea level up to 2000 m. We examined hemoglobin concentration, two markers of oxidative stress (catalase activity and carbonyl concentration) and maximum rate of metabolic respiration at the subcellular level (potential metabolic activity at the electron transport system). The traits were analysed in laboratory acclimated adult male lizards to investigate the adaptive metabolic responses to the variable environmental conditions at the local sampling sites. Potential metabolic activity at the cellular level was measured at four temperatures - 28 °C, 30 °C, 32 °C and 34 °C - covering the range of preferred body temperatures of the species studied. Hemoglobin content, carbonyl concentration and potential metabolic activity did not differ significantly among species. Interspecific differences were found in the catalase activity, Potential metabolic activity increased with temperature in parallel in all five species. The highest response of the metabolic rate with temperature (Q10) and Arrhenius activation energy (Ea) was recorded in the high-mountain species Iberolacerta monticola.

Parasitemia and elevation as predictors of hemoglobin concentration and antioxidant capacity in two sympatric lizards.

Studies which quantify the influence of abiotic factors on physiological variation are paramount to comprehend organismal responses to diverse environments. We studied three physiological aspects of metabolism in two sympatric and ecologically similar European lizard species, Podarcis muralis and Iberolacerta horvathi, across an 830-m elevational gradient. We collected blood samples and tail tips from adult lizards, which were analyzed for parasitemia, hemoglobin concentration, potential metabolic activity and catalase activity. Hemoglobin concentration was higher in males than females and it increased across elevation in one of the studied species - P. muralis. Parasitemia was not an important predictor of the variation in hemoglobin concentration, which suggests that blood parasites do not constraint the aerobic capacity of the lizards. On the other hand, catalase activity reflected increased antioxidant activity in the presence of higher parasitemia, possibly acting as an adaptive mechanism to reduce oxidative stress during immune activation. Potential metabolic activity, as a proxy for maximum respiratory enzymatic capacity, did not differ between species or sexes nor was it affected by elevation or levels of parasitemia. The results provide insight into the relationships between physiological, biotic, and environmental traits in sympatric lizards.

Genotype-environment interactions rule the response of a widespread butterfly to temperature variation.

Understanding how organisms adapt to complex environments is a central goal of evolutionary biology and ecology. This issue is of special interest in the current era of rapidly changing climatic conditions. Here, we investigate clinal variation and plastic responses in life history, morphology and physiology in the butterfly Pieris napi along a pan-European gradient by exposing butterflies raised in captivity to different temperatures. We found clinal variation in body size, growth rates and concomitant development time, wing aspect ratio, wing melanization and heat tolerance. Individuals from warmer environments were more heat-tolerant and had less melanised wings and a shorter development, but still they were larger than individuals from cooler environments. These findings suggest selection for rapid growth in the warmth and for wing melanization in the cold, and thus fine-tuned genetic adaptation to local climates. Irrespective of the origin of butterflies, the effects of higher developmental temperature were largely as expected, speeding up development; reducing body size, potential metabolic activity and wing melanization; while increasing heat tolerance. At least in part, these patterns likely reflect adaptive phenotypic plasticity. In summary, our study revealed pronounced plastic and genetic responses, which may indicate high adaptive capacities in our study organism. Whether this may help such species, though, to deal with current climate change needs further investigation, as clinal patterns have typically evolved over long periods.

Comparing Enchytraeus albidus populations from contrasting climatic environments suggest a link between cold tolerance and metabolic activity.

Basal metabolic activity and freezing of body fluids create reactive oxygen species (ROS) in freeze-tolerant organisms. These sources of ROS can have an additive negative effect via oxidative stress. In cells, antioxidant systems are responsible for removing ROS in order to avoid damage due to oxidative stress. Relatively little is known about the importance of metabolic rate for the survival of freezing, despite a good understanding of several cold tolerance related physiological mechanisms. We hypothesized that low basal metabolism would be selected for in freeze-tolerant organisms where winter survival is important for fitness for two reasons. First, avoidance of the additive effect of ROS production from metabolism and freezing, and second, as an energy-saving mechanism under extended periods of freezing where the animal is metabolically active, but unable to feed. We used the terrestrial oligochaete, Enchytraeus albidus, which is widely distributed from Spain to the high Arctic and compared eight populations originating across a broad geographical and climatic gradient after they had been cold acclimated at 5 °C in a common garden experiment. Cold tolerance (lower lethal temperature: LT50) and the potential metabolic activity (PMA, an estimator of the maximal enzymatic potential of the mitochondrial respiration chain) of eight populations were positively correlated amongst each other and correlated negatively with latitude and positively with average yearly temperature and the average temperature of the coldest month. These results indicate that low PMA in cold tolerant populations is important for survival in extremely cold environments.

The role of metabolism in understanding the altitudinal segregation pattern of two potentially interacting lizards.

Sympatric species from the same ecological guild, that exhibit partial altitudinal segregation, can potentially interact in areas of syntopic occurrence. Besides general species' ecology, physiology can provide important answers about species interactions reflected in altitudinal patterns. Lizards Podarcis muralis and Iberolacerta horvathi exhibit partial altitudinal segregation, while they strongly resemble in overall morphology and ecology (diet, daily and seasonal activity pattern), but show certain degree of physiological dissimilarity. They have similar mean preferred body temperatures and patterns of seasonal and daily variations but differ in the magnitude of seasonal variation. Since an ectotherm metabolism is highly dependent on body temperature, thermoregulation is expected to directly affect their metabolism. We compared metabolic rates of adult males from an area of sympatry, measured under two temperature regimes (20°C and 28°C). Both species increased metabolic rates with temperature in a similar pattern. We also compared electron transport activity from tail tissues which provide values of species' potential metabolic activity (enzymatic capacity). Species clearly differed in potential metabolic activity; I. horvathi attained higher values than P. muralis. No difference was detected in how species exploited this potential (calculated from the ratio of electron transport activity and metabolic rates). However, we observed higher potential metabolic activity I. horvathi which together with the ability to thermoregulate more precisely could represent a higher competitive advantage over P. muralis in thermally more restrictive environments such as higher altitudes. Understanding of metabolism seems to provide valuable information for understanding recent distributional patterns as well as species interactions.

Effects of increased temperature on metabolic activity and oxidative stress in the first life stages of marble trout (Salmo marmoratus).

Climate change may result in future alterations in thermal regime which could markedly affect the early developmental stages of cold water fish due to their expected high sensitivity to increasing temperature. In the present study, the effect of temperature increase of 2, 4 and 6°C on the oxygen consumption rate (R), the activity of respiratory electron transport system (ETS) and oxidative stress have been studied in four developmental stages of the marble trout (Salmo marmoratus)-eyed eggs, yolk-sac larvae and juveniles of 1 and 3 months. Oxygen consumption rate and ETS activity increased with level of development and with temperature in all four stages. ETS/R ratios decreased during development and correlated with temperature in eyed eggs, larvae and juveniles of 1 month, but not in juveniles of 3 months. Low ETS/R ratios at higher temperatures indicate stress response in eyed eggs, the most temperature sensitive developmental stage. Catalase (CAT) and glutathione reductase (GR) activities increased during development, but responded differently to elevated temperature in the different developmental stages. Stress in eyed eggs, caused by higher temperatures, resulted in increased oxygen consumption rate and increased activities of CAT and GR. Larvae were sensitive to increased temperature only at the highest experimental temperature of 16°C. Increased temperature did not stress the metabolism of the juveniles, since they were able to compensate their metabolic activity. The earlier developmental stages of marble trout are thus more sensitive to temperature increase than juveniles and therefore more endangered by higher water temperatures. This is the first report connecting oxygen consumption, ETS activity and ETS/R ratio with the activities of antioxidant enzymes in relation to increased temperature in salmonids.

The thermal tolerance of crayfish could be estimated from respiratory electron transport system activity.

Whether electron transport system (ETS) activity could be used as an estimator of crayfish thermal tolerance has been investigated experimentally. Food consumption rate, respiration rates in the air and water, the difference between energy consumption and respiration costs at a given temperature ('potential growth scope', PGS), and ETS activity of Orconectes limosus and Pacifastacus leniusculus were determined over a temperature range of 5-30°C. All concerned parameters were found to be temperature dependent. The significant correlation between ETS activity and PGS indicates that they respond similarly to temperature change. The regression analysis of ETS activity as an estimator of thermal tolerance at the mitochondrial level and PGS as an indicator of thermal tolerance at the organismic level showed the shift of optimum temperature ranges of ETS activity to the right for 2° in O. limosus and for 3° in P. leniusculus. Thus, lower estimated temperature optima and temperatures of optimum ranges of PGS compared to ETS activity could indicate higher thermal sensitivity at the organismic level than at a lower level of complexity (i.e. at the mitochondrial level). The response of ETS activity to temperature change, especially at lower and higher temperatures, indicates differences in the characteristics of the ETSs in O. limosus and P. leniusculus. O. limosus is less sensitive to high temperature. The significant correlation between PGS and ETS activity supports our assumption that ETS activity could be used for the rapid estimation of thermal tolerance in crayfish species.

Leafhopper males compensate for unclear directional cues in vibration-mediated mate localization.

Ambient noise and transmission properties of the substrate pose challenges in vibrational signal-mediated mating behavior of arthropods, because vibrational signal production is energetically demanding. We explored implications of these challenges in the leafhopper Aphrodes makarovi (Insecta: Hemiptera: Cicadellidae) by exposing males to various kinds of vibrational noise on a natural substrate and challenging them to find the source of the female playback. Contrary to expectations, males exposed to noise were at least as efficient as control males on account of similar searching success with less signaling effort, while playing back male-female duets allowed the males to switch to satellite behavior and locate the target without signaling, as expected. We found altered mitochondrial structure in males with high signaling effort that likely indicate early damaging processes at the cellular level in tymbal muscle, but no relation between biochemical markers of oxidative stress and signaling effort. Analysis of signal transmission revealed ambiguous amplitude gradients, which might explain relatively low searching success, but it also indicates the existence of behavioral adaptations to complex vibrational environments. We conclude that the observed searching tactic, emphasizing speed rather than thorough evaluation of directional cues, may compensate for unclear stimuli when the target is near.

Characteristics and impact of aged coal ash with slag emplaced in a karst cave: the case of Divaska jama, Slovenia.

A mixture of coal bottom ash and slag, with a fraction of fly ash (CAFAS) from steam locomotives, was placed in the cave Divaska jama to delimit and level tourist trails. Emplacement began in 1914 and carried on for several decades. The CAFAS mixed with other cave material gradually changed its structure and appearance. Currently the concentration of some elements in the CAFAS (As, Cu, Hg, Ni, Pb, Zn), and also to a lesser extent in cave sediments (Cr, Cu, Ni), indicates a possibly harmful effect on sediment-associated biota based on ecotoxicological assays. Compared to the cave sediment, the CAFAS contains distinctly different mineral phases and presents a different source of radioactivity. Microbial metabolic activity of CAFAS is low, 0.22 µl O2/gDW h, but higher than that of cave sediment. The present environmental hazards from CAFAS are estimated to be low. Whereas the emplacement of CAFAS was seen initially a long-term solution for waste disposal and management of the cave, it turned out that CAFAS enriches the underground environment with inorganic and organic compounds and disperses pollution into the cave ecosystem. After its removal from the cave, the CAFAS should be investigated thoroughly due to its susceptibility to alteration.

Presence of polyethylene terephthalate (PET) fibers in hyporheic zone alters colonization patterns and seasonal dynamics of biofilm metabolic functioning.

Worldwide, the production of plastics is increasing, and plastic pollution in aquatic environments is a major global concern. Under natural conditions, plastic weathers to smaller pieces called microplastics (MP), which come in various shapes, with fibers often being the most common in freshwater sediments. The hyporheic zone, an ecotone between surface and groundwater, is important for the transport and fate of all MP particles. The main metabolic pathways in rivers take place in the hyporheic zone and are driven by a diverse microbial community. The objective of this study was to investigate in situ whether the presence of PET fibers in riverbed sediments affects patterns of colonization and the seasonal dynamics of microbial metabolic activities in the hyporheic zone. The effects of the presence of PET on microbial metabolism were evaluated in situ over a month (colonization study) and over a year (seasonal study) by measuring total protein content (TPC), and microbial respiration as respiratory electron transport system activity (ETSA) and by community-level physiological profiling (CLPP). Additionally, PET fibers were examined under a scanning electron microscope (SEM), and isotopic analysis (δ13C) of PET was performed after one year of exposure to field conditions. The findings demonstrated that during colonization and biofilm formation, and also over the seasons, the date had a large and significant impact on biofilm growth and activity, while PET presence slightly suppressed microbial biomass (TPC) and respiratory activity (ETSA). Overall microbial activity was repressed in the presence of PET fibers but there was a higher capacity for the utilization of complex synthetic polymer substrates (i.e., Tween 40) which have previously been linked to polluted environments. SEM micrographs showed diverse microbial communities adhering to PET fibers but little surface deterioration. Similarly, isotopic analysis suggested little deterioration of PET fibers after one year of in situ conditions. The study indicated that PET fibers present in riverbed sediments could have impacts on the metabolic functioning in rivers and thus affect their self-cleaning ability.

Critical evaluation of biodegradation studies on synthetic plastics through a systematic literature review.

Increasing amounts of plastic waste in the environment and their fragmentation into smaller particles known as microplastics (particles, <5mm) have raised global concerns due to their persistency in the environment and their potential to act as vectors for harmful substances or pathogenic microorganisms. One possible solution to this problem is biodegradation of plastics by microorganisms. However, the scientific information on plastic-degrading microorganisms is scattered across different scientific publications. We conducted a systematic literature review (SLR) with predefined criteria using the online databases of Scopus and Web of Science to find papers on bacterial biodegradation of synthetic petroleum-based polymers. The aims of this SLR were to provide an updated list of all of the currently known bacteria claimed to biodegrade synthetic plastics, to determine and define the best methods to assess biodegradation, to critically evaluate the existing studies, and to propose directions for future research on polymer biodegradation in support of more rapid development of biodegradation technologies. Most of the bacteria identified here from the 145 reviewed papers belong to the phyla Proteobacteria, Firmicutes and Actinobacteria, and most were isolated from contaminated sites, such as landfill sites. Just under a half of the studies (44%) investigated the biodegradability of polyethylenes and derivates, particularly low-density polyethylenes. The methods used to monitor the biodegradation were mainly scanning electron microscopy and Fourier-transform infrared spectroscopy. We propose that: (1) future research should focus on biodegradation of microplastics arising from the most common pollutants (e.g. polyethylenes); (2) bacteria should be isolated from environments that are permanently contaminated with plastics; and (3) a combination of different observational methods should be used to confirm bacterial biodegradation of these plastics. Finally, when reporting, researchers need to follow standard protocols and include all essential information needed for repetition of the experiments by other research groups.

A new method for early assessment of effects of exposing two non-target crustacean species, Asellus aquaticus and Gammarus fossarum, to pesticides, a laboratory study.

A reliable method is needed for assessing the condition of aquatic animals and their resistance to toxic pollutants. The physiological responses of two freshwater crustaceans, Asellus aquaticus and Gammarus fossarum, following in vitro exposure to two pesticides (atrazine and imidacloprid), were measured by a combination of electron transport system (ETS) activity and respiration (R). Short-term exposure concentrations were selected according to standard toxicity tests and ranged from 0.01 mg L(-1) to 10 mg L(-1). When pesticide concentration was greater than 1 mg l(- 1) (which is below the LC(50) [48 hours] determined for both species), A. aquaticus and G. fossarum responded to short-term exposure with elevated levels of R and/or lower levels of ETS activity. One hour exposure to concentrations of up to 10 mg L(-1) showed an effect in both test species. Laboratory tests confirmed that G. fossarum is more sensitive to short-term pesticide exposure than A. aquaticus. The combination of these two methods provides a useful and effective tool for assessing the general condition of aquatic animals. It also enables to determine toxic effects on freshwater biota of specific or combined pollutants. ETS/R ratio may be used as a quick predictor of effects on organisms exposed to pesticides and other stress factors such as changes in temperature, light, salinity, oxygen concentration and food.

Effects of exposing two non-target crustacean species, Asellus aquaticus L., and Gammarus fossarum Koch., to atrazine and imidacloprid.

The physiological responses of two freshwater crustaceans, Asellus aquaticus L. and Gammarus fossarum Koch., following in vitro exposure to two pesticides were measured. Both species responded to short-term exposure with elevated levels of Respiration and/or lower levels of Electron Transport System (ETS) activity. 1 h exposure to concentrations of up to 10 mg L(-1) showed an effect in both test species. Laboratory tests confirmed that G. fossarum is more sensitive to short-term pesticide exposure than A. aquaticus. ETS/R ratio may be used as a quick predictor of effects on organisms exposed to pesticides.

Modelling the effects of multiple stressors on respiration and microbial biomass in the hyporheic zone using decision trees.

Integrity of freshwater surface- and groundwater ecosystems and their ecological and qualitative status greatly depends on ecological processes taking place in streambed sediments overgrown by biofilm, in the hyporheic zone (HZ). Little is known about the interactions and effects of multiple stressors on biologically driven processes in the HZ. In this study, machine learning (ML) tools were used to provide evidence-based information on how stressors and ecologically important environmental factors interact and drive ecological processes and microbial biomass. The ML technique of decision trees using the J48 algorithm was applied to build models from a data set of 342 samples collected over three seasons at 24 sites within the catchments of five gravel-bed rivers in north-central Slovenia. Catchment-scale land use data and reach-scale environmental features indicating the HZ morphology and physical and chemical characteristics of water were used as predictive variables, while respiration (R) and microbial respiratory electron transport system activity (ETSA) were used as response variables indicating ecological processes and total protein content (TPC) indicating microbial biomass. Separate models were built for two HZ depths: 5-15 cm and 20-40 cm. The models with R as a response variable have the highest predictive performance (67-89%) showing that R is a good indicator of complex environmental gradients. The ETSA and TPC models were less accurate (42-67%) but still provide valuable ecological information. The best model show that temperature when combined with selected water quality elements is an important predictor of R at depth of 5-15 cm. The ETSA and TPC models show the combined effects of temperature, catchment land use and selected water quality elements on both response variables. Overall, this study provides new knowledge on how ecological processes occurring in the HZ respond to catchment and reach-scale variables, and provides evidence-based information about complex interactions between temperature, catchment land use and water quality. These interactions are highly dependent on the selection of the response variable, i.e., each response variable is influenced by a specific combination of predictive environmental variables.

Differences in tolerance to anthropogenic stress between invasive and native bivalves.

Tolerance towards environmental stress has been frequently considered as one of the key determinants of invasion success. However, empirical evidence supporting the assumption that invasive species can better endure unfavorable conditions compared with native species is limited and has yielded opposing results. In this study, we examined the tolerance to different stress conditions (thermal stress and trace metal zinc pollution stress) in two phylogenetically related and functionally similar freshwater bivalve species, the native Anodonta anatina and the invasive Sinanodonta woodiana. We assessed potential differences in response to stress conditions using several cellular response assays: efficiency of the multixenobiotic resistance mechanism, respiration estimate (INT reduction capacity), and enzymatic biomarkers. Our results demonstrated that the invasive species overall coped much better with unfavorable conditions. The higher tolerance of S. woodiana was evident from (i) significantly decreased Rhodamine B accumulation indicating more efficient multixenobiotic resistance mechanism; (ii) significantly higher INT reduction capacity and (iii) less pronounced alterations in the activity of stress-related enzymes (glutathione-S-transferase, catalase) and of a neurotoxicity biomarker (cholinesterase) in the majority of treatment conditions in both stress trials. Higher tolerance to thermal extremes may provide physiological benefit for further invasion success of S. woodiana in European freshwaters, especially in the context of climate change.