A validated LC-MS/MS method for simultaneous determination of key glucocorticoids in animal hair for applications in conservation biology.

A new method for the determination of main glucocorticoids (cortisol, cortisone, and corticosterone) in hair by liquid chromatography-tandem mass spectrometry was developed. Glucocorticoids were extracted from hair shafts using methanol followed by solid-phase extraction. A validation test was performed using hair from three species of wild mammals with different body size (0.2-800 kg), lifestyle (terrestrial, burrowing and arboreal species), social organization (living in herds or solitary), and different predicted type of hair glucocorticoids: European bison (Bison bonasus), European hamster (Cricetus cricetus), and Eurasian red squirrel (Sciurus vulgaris). Regardless of the species evaluated, the method shows good linearity for all analytes accompanied by satisfactory accuracy (91-114%) and precision (RSD < 13%). Depending on the analyte and hair origin, the calculated limits of quantification were between 0.05 and 1.19 ng/mL, which corresponds to 1.28-31.51 pg/mg. Using cortisol and cortisone as examples, we have demonstrated that measuring multiple glucocorticoids simultaneously provides more comprehensive information than solely concentrating on one, thereby contributing to a more balanced and reliable interpretation of the acquired results. However, the utility of cortisol metabolites as markers of stress response in keratinized tissues should be substantiated by additional experimental studies on targeted animals. We posit that this paper could serve as a crucial catalyst to prompt such experiments.

An assessment of non-volant terrestrial vertebrates response to wind farms--a study of small mammals.

The majority of studies on the effects of wind energy development on wildlife have been focused on birds and bats, whereas knowledge of the response of terrestrial, non-flying vertebrates is very scarce. In this paper, the impact of three functioning wind farms on terrestrial small mammal communities (rodents and shrews) and the population parameters of the most abundant species were studied. The study was carried out in southeastern Poland within the foothills of the Outer Western Carpathians. Small mammals were captured at 12 sites around wind turbines and at 12 control sites. In total, from 1200 trap-days, 885 individuals of 14 studied mammal species were captured. There was no difference in the characteristics of communities of small mammals near wind turbines and within control sites; i.e. these types of sites were inhabited by a similar number of species of similar abundance, similar species composition, species diversity (H' index) and species evenness (J') (Pielou's index). For the two species with the highest proportion in the communities (Apodemus agrarius and Microtus arvalis), the parameters of their populations (mean body mass, sex ratio, the proportion of adult individuals and the proportion of reproductive female) were analysed. In both species, none of the analysed parameters differed significantly between sites in the vicinity of turbines and control sites. For future studies on the impact of wind turbines on small terrestrial mammals in different geographical areas and different species communities, we recommend the method of paired 'turbine-control sites' as appropriate for animal species with pronounced fluctuations in population numbers.