Temperature-independent telomere lengthening with age in the long-lived human fish (Proteus anguinus).

Despite a number of studies showing a negative relationship between age and telomere length, the universality of this pattern has been recently challenged, mainly in ectothermic animals exhibiting diverse effects of age on telomere shortening. However, data on ectotherms may be strongly affected by the thermal history of the individuals. We thus investigated the age-related changes in relative telomere length in the skin of a small but long-lived amphibian living naturally in a stable thermal environment over its entire life, allowing comparison with other homeothermic animals like birds and mammals. The present data showed a positive relation between telomere length and individual age, independent of sex and body size. A segmented analysis highlighted a breakpoint in the telomere length-age relationship, suggesting that telomere length reached a plateau at the age of 25 years. Further studies focusing on the biology of animals that live much longer than expected based on body mass will contribute to our better understanding of how ageing processes evolved and may also bring innovation for extending human health span.

Artifical light at night triggers slight transcriptomic effects on melatonin signaling but not synthesis in tadpoles of two anuran species.

The worldwide expansion of artificial light at night (ALAN) is acknowledged as a threat to biodiversity through alterations of the natural photoperiod triggering the disruption of physiological functions. In vertebrates, melatonin production during the dark phase can be decreased or suppressed by nocturnal light as shown in many taxa. But the effect of ALAN at low intensity mimicking light pollution in peri-urban area has never been investigated in amphibians. We filled this gap by studying the impact of low ALAN levels on the expression of genes related to melatonin synthesis and signaling in two anurans (agile frog, Rana dalmatina, and common toad, Bufo bufo). Circadian expression of genes encoding enzymes catalyzing melatonin synthesis (aralkylamine N-acetyltransferase, AANAT and acetylserotonin O-methyltransferase, ASMT) or melatonin receptors (Mel1a, Mel1b and Mel1c) was investigated using RT-qPCR after 23 days of nocturnal exposure to control (< 0.01 lx) or low ALAN (3 lx). We showed that the relative abundance of most transcripts was low in late afternoon and early evening (06 pm and 08 pm) and increased throughout the night in R. dalmatina. However, a clear and ample nocturnal pattern of target gene expression was not detected in control tadpoles of both species. Surprisingly, a low ALAN level had little influence on the relative expression of most melatonin-related genes. Only Mel1c expression in R. dalmatina and Mel1b expression in B. bufo were affected by ALAN. This target gene approach provides experimental evidence that melatonin signaling pathway was slightly affected by low ALAN level in anuran tadpoles.

Microplastics in Freshwater Sediments Impact the Role of a Main Bioturbator in Ecosystem Functioning.

While microplastic transport, fate, and effects have been a focus of studies globally, the consequences of their presence on ecosystem functioning have not received the same attention. With increasing evidence of the accumulation of microplastics at sediment-water interfaces there is a need to assess their impacts on ecosystem engineers, also known as bioturbators, which have direct and indirect effects on ecosystem health. This study investigated the impact of microplastics on the bioturbator Tubifex tubifex alongside any effects on the biogeochemical processes at the sediment-water interface. Bioturbators were exposed to four sediment microplastic concentrations: 0, 700, 7000, and 70000 particles kg-1 sediment dry weight. Though no mortality was present, a significant response to oxidative stress was detected in tubificid worms after exposure to medium microplastic concentration (7000 particles kg-1 sediment dry weight). This was accompanied by a reduction in worm bioturbation activities assessed by their ability to rework sediment and to stimulate exchange water fluxes at the sediment-water interface. Consequently, the contributions of tubificid worms on organic matter mineralization and nutrient fluxes were significantly reduced in the presence of microplastics. This study demonstrated that environmentally realistic microplastic concentrations had an impact on biogeochemical processes at the sediment-water interface by reducing the bioturbation activities of tubificid worms.

Transcriptome-wide deregulation of gene expression by artificial light at night in tadpoles of common toads.

Artificial light at night (ALAN) affects numerous physiological and behavioural mechanisms in various species by potentially disturbing circadian timekeeping systems and modifying melatonin levels. However, given the multiple direct and indirect effects of ALAN on organisms, large-scale transcriptomic approaches are essential to assess the global effect of ALAN on biological processes. Moreover, although studies have focused mainly on variations in gene expression during the night in the presence of ALAN, it is necessary to investigate the effect of ALAN on gene expression during the day. In this study, we combined de novo transcriptome sequencing and assembly, and a controlled laboratory experiment to evaluate the transcriptome-wide gene expression response using high-throughput (RNA-seq) in Bufo bufo tadpoles exposed to ecologically relevant light levels. Here, we demonstrated for the first time that ALAN affected gene expression at night (3.5% and 11% of differentially expressed genes when exposed to 0.1 and 5 lx compared to controls, respectively), but also during the day (11.2% of differentially expressed genes when exposed to 5 lx compared to controls) with a dose-dependent effect. ALAN globally induced a downregulation of genes (during the night, 58% and 62% of the genes were downregulated when exposed to 0.1 and 5 lx compared to controls, respectively, and during the day, 61.2% of the genes were downregulated when exposed to 5 lx compared to controls). ALAN effects were detected at very low levels of illuminance (0.1 lx) and affected mainly genes related to the innate immune system and, to a lesser extend to lipid metabolism. These results provide new insights into understanding the effects of ALAN on organism. ALAN impacted the expression of genes linked to a broad range of physiological pathways at very low levels of ALAN during night-time and during daytime, potentially resulting in reduced immune capacity under environmental immune challenges.

Multiple mating in the context of interspecific hybridization between two Tetramorium ant species.

In eusocial Hymenoptera, haplodiploidy and polyandry may facilitate selection for hybridization. Interspecific hybridization is widespread in ants and can lead to hybrid inviability as well as the formation of new species through hybrid speciation. However, in ants, polyandry is uncommon. By analyzing microsatellite markers on 15 ant workers per colony, we show that the mating system of 28 pure colonies of Tetramorium immigrans, 15 pure colonies of Tetramorium caespitum, and 27 hybrid colonies is a monogyne/polyandrous mating system, with a higher mating rate in T. caespitum (mean = 2.4 males vs. 1.7 in T. immigrans). Hybrid queens, but no hybrid fathers, were deduced from workers' genotypes, in accordance with Haldane's rule extended to haplodiploid organisms, which states that the haploid sex should more often be sterile or inviable. In five colonies, hybridization and multiple mating allowed the simultaneous production of both hybrid and nonhybrid offspring. Although rare, these situations hinted at asymmetrical, larger contributions of T. immigrans vs. T. caespitum males to offspring production. Together, these findings point toward a complex and dynamic mating system in T. immigrans and T. caespitum, and contribute to better understand interspecific hybridization mechanisms and their consequences on genetic and taxonomic diversity. The study of polyandry within a hybrid zone is unprecedented and opens new opportunities to better understand interspecific hybridization mechanisms and their short- to long-term consequences.

Artificial light at night alters the sexual behaviour and fertilisation success of the common toad.

Artificial Light At Night (ALAN) is an emerging pollution, that dramatically keeps on increasing worldwide due to urbanisation and transport infrastructure development. In 2016, it nearly affected 23% of the Earth's surface. To date, all terrestrial and aquatic ecosystems have been affected. The disruption of natural light cycles due to ALAN is particularly expected for nocturnal species, which require dark periods to forage, move, and reproduce. Apart from chiropterans, amphibians contain the largest proportion of nocturnal species among vertebrates exhibiting an unfavourable conservation status in most parts of the world and living in ALAN polluted areas. Despite the growing number of studies on this subject, our knowledge on the direct influence of nocturnal lighting on amphibians is still scarce. To better understand the consequences of ALAN on the breeding component of amphibian fitness, we experimentally exposed male breeding common toads (Bufo bufo) to ecologically relevant light intensities of 0.01 (control), 0.1 or 5 lux for 12 days. At mating, exposed males took longer than controls to form an amplexus, i.e. to pair with a female, and broke amplexus before egg laying, while controls never did. These behavioural changes were associated with fitness alteration. The fertilisation rate of 5 lux-exposed males was reduced by 25%. Salivary testosterone, which is usually correlated with reproductive behaviours, was not altered by ALAN. Our study demonstrates that ALAN can affect the breeding behaviour of anuran species and reduce one component of their fitness. Given the growing importance of ALAN, more work is needed to understand its long-term consequences on the behaviour and physiology of individuals. It appears essential to identify deleterious effects for animal populations and propose appropriate management solutions in an increasingly brighter world.

The role of associative learning process on the response of fledgling great tits (Parus major) to mobbing calls.

When they detect a predator, many species emit anti-predator vocalizations. In some cases, they emit mobbing calls, which are associated with the caller approaching and harassing the predator while attracting others to join it. Surprisingly, although mobbing has been widely reported in adults of numerous species, there has been no test of the role of learning in mobbing call recognition, especially during ontogeny. Here, we exposed wild great tit (Parus major) nestlings to playbacks of an unthreatening novel sound either associated with conspecific mobbing calls (experimental treatment) or with another unthreatening novel sound (control treatment). We then tested them as nestlings and fledglings to see how they respond to the novel sound compared to conspecific mobbing calls. Results revealed that fledglings in the experimental treatment behaved similarly to conspecific mobbing calls and the novel sound associated with conspecific mobbing calls. Because mobbing efficiency is often linked to interspecific communication, associative learning should be used by heterospecifics as mobbing calls recognition mechanism. Regardless of treatment during the nestling phase, fledglings always were sensitive to the playback of conspecific mobbing calls. However, fledglings from the control group were more likely to approach the loudspeaker than those from the experimental group when mobbing calls were played suggesting that overexposure during the nestling phase altered mobbing learning. Overall, these results suggest that learning could play a role in the recognition of calls, like heterospecific mobbing calls, when paired with conspecific mobbing, and that mobbing is perceived as a threatening stimulus from a very young age.

Doping for sex: Bad for mitochondrial performances? Case of testosterone supplemented Hyla arborea during the courtship period.

Sexual selection has been widely explored from numerous perspectives, including behavior, ecology, and to a lesser extent, energetics. Hormones, and specifically androgens such as testosterone, are known to trigger sexual behaviors. Their effects are therefore of interest during the breeding period. Our work investigates the effect of testosterone on the relationship between cellular bioenergetics and contractile properties of two skeletal muscles involved in sexual selection in tree frogs. Calling and locomotor abilities are considered evidence of good condition in Hyla males, and thus server as proxies for male quality and attractiveness. Therefore, how these behaviors are powered efficiently remains of both physiological and behavioral interest. Most previous research, however, has focused primarily on biomechanics, contractile properties or mitochondrial enzyme activities. Some have tried to establish a relationship between those parameters but to our knowledge, there is no study examining muscle fiber bioenergetics in Hyla arborea. Using chronic testosterone supplementation and through an integrative study combining fiber bioenergetics and contractile properties, we compared sexually dimorphic trunk muscles directly linked to chronic sound production to a hindlimb muscle (i.e. gastrocnemius) that is particularly adapted for explosive movement. As expected, trunk muscle bioenergetics were more affected by testosterone than gastrocnemius muscle. Our study also underlines contrasted energetic capacities between muscles, in line with contractile properties of these two different muscle phenotypes. The discrepancy of both substrate utilization and contractile properties is consistent with the specific role of each muscle and our results are elucidating another integrative example of a muscle force-endurance trade-off.

Effects of traffic noise on tree frog stress levels, immunity, and color signaling.

During the last decade, many studies have focused on the detrimental effects of noise pollution on acoustic communication. Surprisingly, although it is known that noise exposure strongly influences health in humans, studies on wildlife remain scarce. In order to gain insight into the consequences of traffic noise exposure, we experimentally manipulated traffic noise exposure as well as the endocrine status of animals to investigate physiological and phenotypic consequences of noise pollution in an anuran species. We showed that noise exposure increased stress hormone level and induced an immunosuppressive effect. In addition, both traffic noise exposure and stress hormone application negatively impacted H. arborea vocal sac coloration. Moreover, our results suggest profound changes in sexual selection processes because the best quality males with initial attractive vocal sac coloration were the most impacted by noise. Hence, our study suggests that the recent increases in anthropogenic noise worldwide might affect a broader range of animal species than previously thought, because of alteration of visual signals and immunity. Generalizing these results to other taxa is crucial for the conservation of biodiversity in an increasingly noisy world.

All or nothing: Survival, reproduction and oxidative balance in Spotted Wing Drosophila (Drosophila suzukii) in response to cold.

Winter severity and overwintering capacity are key ecological factors in successful invasions, especially in ectotherms. The integration of physiological approaches into the study of invasion processes is emerging and promising. Physiological information describes the mechanisms underlying observed survival and reproductive capacities, and it can be used to predict an organism's response to environmental perturbations such as cold temperatures. We investigated the effects of various cold treatments on life history and physiological traits of an invasive pest species, Drosophila suzukii, such as survival, fertility and oxidative balance. This species, a native of temperate Asian areas, is known to survive where cold temperatures are particularly harsh and has been recently introduced into Europe and North America. We found that cold treatments had a strong impact on adult survival but no effect on female's fertility. Although only minor changes were observed after cold treatment on studied physiological traits, a strong sex-based difference was observed in both survival and physiological markers (antioxidant defences and oxidative markers). Females exhibited higher survival, reduced oxidative defences, less damage to nucleic acids, and more damage to lipids. These results suggest that D. suzukii relies on a pathway other than oxidative balance to resist cold injury. Altogether, our results provide information concerning the mechanisms of successful invasion by D. suzukii. These findings may assist in the development of population models that predict the current and future geographic ranges of this species.

Impact of cadmium on the ecdysteroids production in Gammarus fossarum.

Gammarus fossarum is an important test organism which is currently used as a bio-indicator as well as in ecotoxicological tests. Nevertheless, data on ecdysteroids in endocrine toxicity test are not yet available for these species, despite its crucial role in molting and reproduction. In the present paper, ecdysteroids concentrations were studied during the molt cycle (in females) and embryonic development in G. fossarum (Crustacea, Amphipoda) in order to propose an ecdysteroids toxicity test. Ecdysteroids levels in G. fossarum showed a single peak during premolt at stage Dl-D2. In embryos, ecdysteroids levels progressively increased over stages 3 and 4, with peak levels at stage 4. A Cadmium toxicity test was proposed to examine if the molting and embryogenesis disturbances previously observed after cadmium exposure (Geffard et al. 2010) could be attributed to changes in ecdysteroids titers. Exposure to the different cadmium concentrations (3; 9; 300; 900 µg/l) increased ecdysteroids secretion by Y-organs in vitro, but it had no significant effect on exposed embryos (in vivo). Based on previous findings, we are led to conclude that the molting impairments in cadmium-exposed females of G. fossarum is connected to the changes in ecdysteroids concentrations.

Oxidative phosphorylation efficiency, proton conductance and reactive oxygen species production of liver mitochondria correlates with body mass in frogs.

Body size is a central biological parameter affecting most biological processes (especially energetics) and the mitochondrion is a key organelle controlling metabolism and is also the cell's main source of chemical energy. However, the link between body size and mitochondrial function is still unclear, especially in ectotherms. In this study, we investigated several parameters of mitochondrial bioenergetics in the liver of three closely related species of frog (the common frog Rana temporaria, the marsh frog Pelophylax ridibundus and the bull frog Lithobates catesbeiana). These particular species were chosen because of their differences in adult body mass. We found that mitochondrial coupling efficiency was markedly increased with animal size, which led to a higher ATP production (+70%) in the larger frogs (L. catesbeiana) compared with the smaller frogs (R. temporaria). This was essentially driven by a strong negative dependence of mitochondrial proton conductance on body mass. Liver mitochondria from the larger frogs (L. catesbeiana) displayed 50% of the proton conductance of mitochondria from the smaller frogs (R. temporaria). Contrary to our prediction, the low mitochondrial proton conductance measured in L. catesbeiana was not associated with higher reactive oxygen species production. Instead, liver mitochondria from the larger individuals produced significantly lower levels of radical oxygen species than those from the smaller frogs. Collectively, the data show that key bioenergetics parameters of mitochondria (proton leak, ATP production efficiency and radical oxygen species production) are correlated with body mass in frogs. This research expands our understanding of the relationship between mitochondrial function and the evolution of allometric scaling in ectotherms.

Thyroid status affects membranes susceptibility to free radicals and oxidative balance in skeletal muscle of Muscovy ducklings (Cairina moschata).

Thyroid hormones (TH) are major contributor to oxidative stress in mammals because they (1) stimulate reactive oxygen species generation (ROS), (2) impair antioxidant defenses, and (3) increase the susceptibility to free radicals of most tissues. Unlike mammals, THs seem to diminish mitochondrial ROS while they have limited effect on the antioxidant machinery in birds. However, how THs modify the susceptibility to ROS has never been explored in an avian model, and very little is known about their effect on oxidative balance in birds. Therefore, the objective of our study was to examine the effect of chronic pharmacological hypo- and hyperthyroidism on (i) the susceptibility of mitochondrial membranes to ROS; and (ii) the level of oxidative stress assessed by measuring oxidative damage to lipids, nucleic acids and proteins in the gastrocnemius muscle of ducklings. We show that hypothyroidism had no effect on the susceptibility of mitochondrial membranes to free radicals. Hypothyroid ducklings had lower oxidized lipids (-31%) and DNA (-25%) but a similar level of protein carbonylation relative to controls. Conversely, mitochondrial membranes of hyperthyroid ducklings exhibited higher unsaturation (+12%) and peroxidation (+31%) indexes than in controls indicating a greater susceptibility to free radicals. However, hyperthyroid ducklings exhibited more oxidative damages on proteins (+67%) only, whereas lipid damages remained unchanged, and there was a slight reduction (-15%) in damages to DNA compared to euthyroid controls. Our results indicate that birds and mammals present fundamental differences in their oxidative stress response to thyroid status.