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  • Publication
    Accès libre
    Sensitive and selective quantification of free and total malondialdehyde in plasma using UHPLC-HRMS
    Quantification of malondialdehyde (MDA) as a marker of lipid peroxidation is relevant for many research fields. We describe a new sensitive and selective method to measure free and total plasmatic MDA, using derivatization with 2,4-dinitrophenylhydrazine (DNPH) and ultrahigh performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS). Free and total MDA were extracted from minute sample amounts (10 μL) using acidic precipitation and alkaline hydrolysis followed by acidic precipitation, respectively. Derivatization was completed within 10 min at room temperature, and the excess DNPH discarded by liquid-liquid extraction. Quantification was achieved by internal standardization using dideuterated MDA as internal standard. The method lowest limit of quantification (LLOQ) was 100 nM and linearity spanned > 3 orders of magnitude. Intra- and inter-day precisions for total MDA were 2.9% and 3.0%, respectively, and those for free MDA were 12.8% and 24.9%, respectively. Accuracy was 101% and 107% at low and high concentrations, respectively. In human plasma, free MDA levels were 120 nM (SD 36.26) and total MDA levels were 6.7 μM (SD 0.46). In addition, we show the applicability of this method to measure MDA plasma levels from a variety of animal species, making it invaluable to scientists in various fields.
  • Publication
    Accès libre
    Antioxidant allocation modulates sperm quality across changing social environments
    In promiscuous species, male reproductive success depends on their ability to mate with fertile females and on the fertilizing ability of their sperm. In such species, theory predicts that, owing to a trade-off between pre- and post-copulatory reproductive traits, males with lesser access to females should increase resource investment into those sperm traits that enhance fertilization success–usually referred to as ejaculate quality. This prediction has been validated in several taxa, yet studies on the physiological mechanisms modulating ejaculate quality are lacking. Sperm cells are highly vulnerable to oxidative stress, which impairs male fertility. Therefore, males that better protect their sperm from oxidative stress are expected to achieve higher ejaculate quality. Based on theoretical expectations, and since social dominance is a major determinant of mating opportunity, we predicted that subordinate males should invest more into the antioxidant protection of their sperm in order to achieve higher ejaculate quality. We maintained 60 male and 60 female wild-caught house sparrows Passer domesticus in outdoor aviaries, where we experimentally manipulated male social status to test our predictions. We measured cellular oxidative stress and enzymatic antioxidant activity in blood and sperm both before and after manipulating social ranks. Before manipulating the social status, we found that ejaculate viability correlated with oxidative stress level in sperm, with dominant males producing more oxidized and less viable ejaculates. Further, males at the lower end of the hierarchy produced ejaculates of similar quality to those of dominant males, suggesting that restricted access to resources might limit male reproductive strategies. After experimentally manipulating the social status, males matched their ejaculate quality to their new rank, while increases in antioxidant investment into ejaculates paralleled increases in ejaculate viability. Oxidative stress has been proposed as a general constraint to the evolution of life histories. Our results highlight oxidative stress and strategic antioxidant allocation as important proximate physiological mechanisms underlying male reproductive strategies.
  • Publication
    Métadonnées seulement
    Reproductive effort and oxidative stress: effects of offspring sex and number on the physiological state of a long-lived bird
    (2017-2-13)
    Merkling, Thomas
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    Blanchard, Pierrick
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    Chastel, Olivier
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    Vallat, Armelle
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    Hatch, Scott A.
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    Danchin, Etienne
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    1. Individuals must trade-off between energetically costly activities to maximize their fitness. However, the underlying physiological mechanism remains elusive. Oxidative stress, the imbalance between reactive oxygen species production and antioxidant and/or repair activities, has been suggested to underlie life-history trade-offs: greater investment in reproduction supposedly generating higher oxidative damage, thus reducing life span. 2. While most studies used natural or experimental variation in offspring number to examine how reproduction affects oxidative stress, none studied the impact of offspring sex, although it could influence physiological costs and fitness, if the sexes differ in terms of energetic cost. 3. Here, we aim at further understanding how reproduction (in terms of offspring sex, experimentally manipulated and number, not manipulated) influences oxidative stress in a wild seabird, where sons are energetically costlier than daughters. We did so by conducting a chick fostering experiment (to disentangle foster and produced sex ratio) and using four oxidative stress markers plus baseline corticosterone. 4. First, the results suggest that individual physiological state before laying modulates upcoming reproductive effort. Individuals with higher pre-laying baseline corticosterone and lower antioxidant activity, estimated by their superoxide dismutase activity, subsequently invested more in reproduction, estimated by the cumulative number of days spent rearing chicks. Hence, it seems that only individuals that could afford to invest heavily in reproduction did so. 5. Then, we examined the effects of reproductive effort on individual physiological state at the end of the breeding season. Higher reproductive effort seemed to imply higher physiological costs. Oxidative stress, estimated by the ratio of oxidized over reduced glutathione, increased with more male-biased foster sex ratio among mothers but not among fathers, whereas baseline corticosterone did so in both sexes. Similarly, lipid oxidative damage to red blood cells increased with increasing cumulative number of days spent rearing chicks. 6. Our study provides the first evidence that brood sex ratio variation can affect oxidative balance, potentially in a sex-specific way, although more studies are needed to understand whether the observed physiological costs could lead to fitness costs. It also highlights the need to consider sex ratio in future studies investigating the role of oxidative stress in life-history trade-offs.
  • Publication
    Métadonnées seulement
    Alternative reproductive tactics, sperm mobility, and oxidative stress in Carollia perspicillata (Seba's short-tailed bat)
    (2017-1-1)
    Fasel, Nicolas Jean
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    Wesseling, Charlotte
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    Fernandez, Ahana Aurora
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    Vallat, Armelle
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    ; ;
    Richner, Heinz
    In social systems with alternative reproductive tactics, sneakers face a stronger sperm competition than harem males, and hence are predicted to allocate more resources to ejaculates. Antioxidants can protect sperm against oxidative stress, and thus their allocation to the ejaculate may depend on social status. In this study on the frugivorous bat Carollia perspicillata, we assessed for harem and sneaker males, four sperm mobility traits, blood and ejaculate markers of the redox balance, and the ejaculate to blood ratios of the redox markers. Under higher sperm competition, sneaker males should allocate proportionally more antioxidant resources to the protection of sperm than harem males. In contrast, harem males should favour pre-copulatory functions, which comprise the protection of blood. We found significantly higher sperm velocity and sperm survival in sneakers. There was no correlation between sperm mobility and sperm enzymatic antioxidant activity or ejaculate levels of lipid peroxidation (oxidative damage). Ejaculate levels of lipid peroxidation and sperm survival showed a significantly positive correlation, which could be attributed to the role of reactive oxygen species for sperm capacitation. Harem and sneaker males showed similar levels of redox balance markers in ejaculate and blood. However, harem males showed a higher ratio of oxidized over reduced glutathione in blood, which may indicate higher cellular stress due to higher metabolism. Overall, our findings suggest that sneakers of C. perspicillata compensate for a stronger sperm competition by higher sperm mobility.
  • Publication
    Métadonnées seulement
    Corticosterone: effects on feather quality and deposition into feathers
    (2015-2-1)
    Jenni-Eiermann, Susanne
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    Vallat, Armelle
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    Jenni, Lukas
    The concentration of the glucocorticoid hormone corticosterone (CORT) is increasingly used in ecology and conservation biology as an integrated measure of the historical record of an individual's hypothalamo–pituitary–adrenal (HPA) activity during feather growth. However, where and how CORT is incorporated in feathers is incompletely known. * We therefore examined whether CORT is reliably measured with an enzyme immunoassay, where CORT is incorporated in the feather and where it affects feather quality, and whether CORT incorporation is related to plasma CORT levels, feather growth rate and melanin pigmentation. * During the regrowth of plucked tail feathers, we injected pigeons with tritium-labelled CORT, and implanted a CORT-releasing pellet to increase plasma CORT concentration for about 3 days. In feather segments, we measured labelled CORT (DPM3H) and we quantified CORT with an enzyme immunoassay EIA (CORTEIA) and double-checked the results with ultra-high performance liquid chromatography–tandem mass spectrometry (LCMS) (CORTMS). * Administered CORT affected feather structure and colour at the very base of the feather (epidermal collar, ramogenic zone) and reduced growth rate. In contrast, incorporation of CORT into the feather happened mainly in the blood quill, as shown with all three methods (DPM3H, CORTEIA and CORTMS). * Incorporation of CORT into feathers was only roughly proportional to plasma concentration, proportional to feather growth rate and increased with melanin pigmentation. * Measuring CORT in feather is a way to reveal past events of increased stress during feather growth in birds.
  • Publication
    Accès libre
    Badge Size Reflects Sperm Oxidative Status within Social Groups in the House Sparrow Passer domesticus
    The phenotype-linked fertility hypothesis proposes that male ornaments reflect male fertility. Male ornaments could honestly signal sperm quality due to the high susceptibility of sperm to free radicals on the one hand and the negative impact of oxidative stress on ornament elaboration on the other hand. Thus, only males with superior antioxidant defences could bear the cost of more elaborated sexual ornaments without suffering adverse fitness costs. Yet, in species where males experience differential access to fertile females, a trade-off emerges between investing into traits favouring mating opportunities (e.g. secondary sexual ornaments, social dominance, mate-guarding behaviours, etc.) or into traits favouring sperm competitive ability (e.g. sperm numbers and quality). When male sexual ornaments promote greater access to fertile females, a negative relationship can then be predicted between ornamentation and sperm quality. We tested the latter hypothesis and the phenotype-linked fertility hypothesis in wild House Sparrows Passer domesticus by exploring the relationships between sperm quality, melanin-based ornamentation, and redox status in blood and sperm. We found no correlation between badge size and sperm swimming performance. However, we found that within a social group, large-badged males better protect their ejaculates from oxidative stress, and thus produce less oxidized ejaculates. Additionally, we found that badge size did not reflect social dominance, and thus the protection of the ejaculate is independent of males’ ability to monopolize resources. Our results suggest that badge size might reflect male investment into the antioxidant protection of their sperm relative to a given social environment, and thus females may accrue both direct and indirect benefits by mating with large-badged males producing less oxidized ejaculates.