Non-lethal sampling for assessment of mitochondrial function does not affect metabolic rate and swimming performance.

A fundamental issue in the metabolic field is whether it is possible to understand underlying mechanisms that characterize individual variation. Whole-animal performance relies on mitochondrial function as it produces energy for cellular processes. However, our lack of longitudinal measures to evaluate how mitochondrial function can change within and among individuals and with environmental context makes it difficult to assess individual variation in mitochondrial traits. The aims of this study were to test the repeatability of muscle mitochondrial metabolism by performing two biopsies of red muscle, and to evaluate the effects of biopsies on whole-animal performance in goldfish Carassius auratus. Our results show that basal mitochondrial respiration and net phosphorylation efficiency are repeatable at 14-day intervals. We also show that swimming performance (optimal cost of transport and critical swimming speed) was repeatable in biopsied fish, whereas the repeatability of individual oxygen consumption (standard and maximal metabolic rates) seemed unstable over time. However, we noted that the means of individual and mitochondrial traits did not change over time in biopsied fish. This study shows that muscle biopsies allow the measurement of mitochondrial metabolism without sacrificing animals and that two muscle biopsies 14 days apart affect the intraspecific variation in fish performance without affecting average performance of individuals. This article is part of the theme issue 'The evolutionary significance of variation in metabolic rates'.

Wild versus lab house mice: Effects of age, diet, and genetics on molar geometry and topography.

Molar morphology is shaped by phylogenetic history and adaptive processes related to food processing. Topographic parameters of the occlusal surface, such as sharpness and relief, can be especially informative regarding diet preferences of a species. The occlusal surface can however be deeply modified by wear throughout an animal's life, potentially obliterating other signals. Age being difficult to assess in wild populations, especially small rodents, experimental studies of wear through age in laboratory populations may constitute a powerful way to assess its impact on molar geometry and topography, and to validate descriptors of molar morphology that could mitigate this issue. Molar morphology was therefore quantified using 3D geometric morphometrics and topographic estimates in four groups of house mice: wild-trapped mice, lab-bred offspring of these wild mice, typical laboratory mice, and their hybrids. Three descriptors of the molar morphology were considered: the surface of the whole molar row, the surface of the first upper molar, and a truncated template of the first upper molar mimicking advanced wear. Increasing wear with age was demonstrated in the different groups, with a more pronounced effect in the wild-trapped population. The geometry of the molar row is not only modified by wear, but also by the relative position of the late developing molars on the jaw due to loading during mastication. As a consequence, the alignment of the molars is modified in wild mice, showing a qualitative difference between wild animals and their lab-bred offspring. Results obtained from the lab should thus be transferred with caution to the interpretation of differences in wild populations. Topographic estimates computed for the first upper molar seems to provide more stable parameters than those based on the whole molar row, because issues related to non-planar occlusal surface along the molar row are discarded. The truncated template was proven efficient in discarding the wear effect to focus on genetic differences, allowing an efficient characterization of the hybridization signature between wild and lab mice. Dominance of the wild phenotype for the first molar shape supports that the lab strain evolved in a context of relaxation of the selective pressures related to nutrition.

INTRAMUSCULAR ADMINISTRATION OF KETAMINE-MEDETOMIDINE ASSURES STABLE ANAESTHESIA NEEDED FOR LONG-TERM SURGERY IN THE ARGENTINE TEGU SALVATOR MERIANAE.

To define a protocol of anesthesia for long-duration invasive surgery in a lizard, eight young adult Argentine tegus ( Salvator merianae) of mean body weight 3.0 kg (interquartile range [IQR] 3.40-2.65) were anesthetized with a mixture of ketamine (K) and medetomidine (M) at 19°C, injected intramuscularly and equally distributed in the four limbs. As the experimental surgery procedure required a prolonged deep anesthesia with a good myorelaxation (between 16 and 21 hr), reinjections were required and reflexes were checked during surgery. Times for anesthetic induction, anesthetic reinjection, and recovery periods were recorded for five different combinations of ketamine-medetomidine: 1) 66 mg/kg K + 100 µg/kg M; 2) 80 mg/kg K + 100 µg/kg M; 3) 100 mg/kg K + 130 µg/kg M; 4) 125 mg/kg K + 200 µg/kg M; and 5) 150 mg/kg K + 200 µg/kg M. The effect on the recovery speed of the postoperative atipamezole injection was also evaluated. The median induction time was 30 (IQR 35-27.5) min with no statistical difference between all the concentrations tested. The first reinjection of half a dose was administered after a mean of 5 hr (5.64 hr, IQR 5.95-4.84) as were the subsequent reinjections of a quarter dose (3.99 hr, IQR 5.98-3.23). Intramuscular administration of the ketamine-medetomidine combination is a simple, rapid, and efficient anesthesia for long-term surgery (>12 hr). A mix of 100 mg/kg ketamine and 200 µg/kg medetomidine, with reinjections every 4 hr of half a dose of the previous injection can maintain a good quality of anesthesia for at least 16 hr. The injection of atipamezole after the surgery reverses the effects of medetomidine and permits a reduction of the recovery period.

Bioconcentration of (15)N-tamoxifen at environmental concentration in liver, gonad and muscle of Danio rerio.

Pharmaceutical compounds (PCs) are ubiquitous in aquatic ecosystems. In addition to the direct ecotoxicological risk presented by certain PCs, others can accumulate inside organisms and along trophic webs, subsequently contaminating whole ecosystems. We studied the bioconcentration of a bioaccumulative PC already found several times in the environment: tamoxifen. To this end, we exposed Danio rerio for 21d to (15)N-tamoxifen concentrations ranging from 0.1 to 10µg/L and used an analytic method based on stable isotopes to evaluate the tamoxifen content in these organisms. The evolution of the (15)N/(14)N ratio was thus measured in liver, muscle and gonads of exposed fish compared to control fish. We succeeded in quantifying (15)N-tamoxifen bioconcentrations at all the exposure concentrations tested. The highest bioconcentration factors of tamoxifen measured were 14,920 in muscle, 73,800 in liver and 85,600 in gonads of fish after 21d exposure at a nominal concentration of 10µg/L. However, these bioconcentration factors have to be considered as maximal values (BCFMAX). Indeed, despite its proven stability, tamoxifen can be potentially partially degraded during experiments. We now need to refine these results by using a direct analytic method (i.e. LC-MS/MS).