Measurement of Fecal Testosterone Metabolites in Mice: Replacement of Invasive Techniques

Testosterone is the main reproductive hormone in male vertebrates and conventional methods to measure testosterone rely on invasive blood sampling procedures. Here, we aimed to establish a non-invasive alternative by assessing testosterone metabolites (TMs) in fecal and urinary samples in mice. We performed a radiometabolism study to determine the effects of daytime and sex on the metabolism and excretion pattern of radiolabeled TMs. We performed physiological and biological validations of the applied EIA to measure TMs and assessed diurnal fluctuations in TM excretions in male and female mice and across strains. We found that males excreted significantly more radiolabeled TMs via the feces (59%) compared to females (49.5%). TM excretion patterns differed significantly between urinary and fecal samples and were affected by the daytime of ³H-testosterone injection. Overall, TM excretion occurred faster in urinary than fecal samples. Peak excretion of fecal TMs occurred after 8 h when animals received the 3H-testosterone in the morning, or after 4 h when they received the 3H-testosterone injection in the evening. Daytime had no effect on the formed TMs; however, males and females formed different types of TMs. As expected, males showed higher fecal TM levels than females. Males also showed diurnal fluctuations in their TM levels but we found no differences in the TM levels of C57BL/6J and B6D2F1 hybrid males. Finally, we successfully validated our applied EIA (measuring 17β-hydroxyandrostane) by showing that hCG (human chorionic gonadotropin) administration increased TM levels, whereas castration reduced them. In conclusion, our EIA proved suitable for measuring fecal TMs in mice. Our non-invasive method to assess fecal TMs can be widely used in various research disciplines like animal behavior, reproduction, animal welfare, ecology, conservation, and biomedicine

Testing the effect of medical positive reinforcement training on salivary cortisol levels in bonobos and orangutans

The management of captive animals has been improved by the establishment of positive reinforcement training as a tool to facilitate interactions between caretakers and animals. In great apes, positive reinforcement training has also been used to train individuals to participate in simple medical procedures to monitor physical health. One aim of positive reinforcement training is to establish a relaxed atmosphere for situations that, without training, might be very stressful. This is especially true for simple medical procedures that can require animals to engage in behaviours that are unusual or use unfamiliar medical devices that can be upsetting. Therefore, one cannot exclude the possibility that the training itself is a source of stress. In this study, we explored the effects of medical positive reinforcement training on salivary cortisol in two groups of captive ape species, orangutans and bonobos, which were familiar to this procedure. Furthermore, we successfully biologically validated the salivary cortisol assay, which had already been validated for bonobos, for orangutans. For the biological validation, we found that cortisol levels in orangutan saliva collected during baseline conditions were lower than in samples collected during three periods that were potentially stressful for the animals. However, we did not find significant changes in salivary cortisol during medical positive reinforcement training for either bonobos or orangutans. Therefore, for bonobos and orangutans with previous exposure to medical PRT, the procedure is not stressful. Thus, medical PRT provides a helpful tool for the captive management of the two species

Measurement of fecal glucocorticoid metabolites and evaluation of udder characteristics to estimate stress after sudden dry-off in dairy cows with different milk yields

Sudden dry-off is an established management practice in the dairy industry. But milk yield has been increasing continuously during the last decades. There is no information whether the dry-off procedure, which often results in swollen and firm udders, causes stress, particularly in high-producing dairy cows. Therefore, we evaluated the effect of a sudden dry-off on extramammary udder pressure and the concentration of fecal glucocorticoid metabolites (i.e., 11,17-dioxoandrostane, 11,17-DOA) as an indirect stress parameter. Measurements were carried out within the last week before dry-off and until 9d after dry-off considering 3 groups of milk yield (i.e., low: <15 kg/d, medium: 15-20 kg/d, and high: >20 kg/d). Udder pressure increased in all yield groups after dry-off, peaked at d 2 after dry-off and decreased afterwards. Pressures were highest in high-yielding cows and lowest in low-yielding cows. But only in high-yielding cows was udder pressure after dry-off higher than before dry- off. Baseline 11,17-DOA concentrations depended on milk yield. They were highest in low-yielding (121.7 ± 33.3 ng/g) and lowest in high-yielding cows (71.1 ± 30.0 ng/g). After dry-off, 11,17-DOA increased in all yield groups and peaked at d 3. Whereas in medium- and high-yielding cows 11,17-DOA levels differed significantly from their respective baseline during the whole 9-d measuring period, low-yielding cows showed elevated 11,17-DOA levels only on d 3 after dry-off. However, especially the increase in 11,17-DOA after dry-off between the 3 yield groups was considerably different. Mean 11,17-DOA increase from baseline to d 3 was highest in high-yielding cows (129.1%) and considerably lower in low-yielding cows (40.1%). The highest fecal 11,17-DOA concentrations were measured on d 3 after dry-off, indicating that the stress was most intense on d 2, which is due to an 18-h time lag; at about the same time, udder pressure peaked. Our results showed a negligible effect of a sudden dry-off on low-yielding cows. High-yielding cows, however, faced high extramammary pressures and increased glucocorticoid production. Considering animal welfare aspects, a review of the current dry-off strategies might be warranted

Comparing plasma and faecal measures of steroid hormones in Adelie penguins Pygoscelis adeliae

Physiological measurements of both stress and sex hormones are often used to estimate the consequences of natural or human-induced change in ecological studies of various animals. Different methods of hormone measurement exist, potentially explaining variation in results across studies; methods should be cross-validated to ensure that they correlate. We directly compared faecal and plasma hormone measurements for the first time in a wild free-living species, the Adelie penguin (Pygoscelis adeliae). Blood and faecal samples were simultaneously collected from individual penguins for comparison and assayed for testosterone and corticosterone (or their metabolites). Sex differences and variability within each measure, and correlation of values across measures were compared. For both hormones, plasma samples showed greater variation than faecal samples. Males had higher mean corticosterone concentrations than females, but the difference was only statistically significant in faecal samples. Plasma testosterone, but not faecal testosterone, was significantly higher in males than females. Correlation between sample types was poor overall, and weaker in females than in males, perhaps because measures from plasma represent hormones that are both free and bound to globulins, whereas measures from faeces represent only the free portion. Faecal samples also represent a cumulative measure of hormones over time, as opposed to a plasma ‘snapshot’ concentration. Our data indicate that faecal sampling appears more suitable for assessing baseline hormone concentrations, whilst plasma sampling may best define immediate responses to environmental events. Consequently, future studies should ensure that they select the most appropriate matrix and method of hormone measurement to answer their research questions

Connecting speeds, directions and arrival times of 22 coronal mass ejections from the Sun to 1 AU

Forecasting the in situ properties of coronal mass ejections (CMEs) from remote images is expected to strongly enhance predictions of space weather, and is of general interest for studying the interaction of CMEs with planetary environments. We study the feasibility of using a single heliospheric imager (HI) instrument, imaging the solar wind density from the Sun to 1 AU, for connecting remote images to in situ observations of CMEs. We compare the predictions of speed and arrival time for 22 CMEs (in 2008-2012) to the corresponding interplanetary coronal mass ejection (ICME) parameters at in situ observatories (STEREO PLASTIC/IMPACT, Wind SWE/MFI). The list consists of front- and backsided, slow and fast CMEs (up to $2700 \: km \: s^{-1}$). We track the CMEs to $34.9 \pm 7.1$ degrees elongation from the Sun with J-maps constructed using the SATPLOT tool, resulting in prediction lead times of $-26.4 \pm 15.3$ hours. The geometrical models we use assume different CME front shapes (Fixed-$\Phi$, Harmonic Mean, Self-Similar Expansion), and constant CME speed and direction. We find no significant superiority in the predictive capability of any of the three methods. The absolute difference between predicted and observed ICME arrival times is $8.1 \pm 6.3$ hours ($rms$ value of 10.9h). Speeds are consistent to within $284 \pm 288 \: km \: s^{-1}$. Empirical corrections to the predictions enhance their performance for the arrival times to $6.1 \pm 5.0$ hours ($rms$ value of 7.9h), and for the speeds to $53 \pm 50 \: km \: s^{-1}$. These results are important for Solar Orbiter and a space weather mission positioned away from the Sun-Earth line.Comment: 19 pages, 13 figures, accepted for publication in the Astrophysical Journa

Multiwavelength Study on Solar and Interplanetary Origins of the Strongest Geomagnetic Storm of Solar Cycle 23

We study the solar sources of an intense geomagnetic storm of solar cycle 23 that occurred on 20 November 2003, based on ground- and space-based multiwavelength observations. The coronal mass ejections (CMEs) responsible for the above geomagnetic storm originated from the super-active region NOAA 10501. We investigate the H-alpha observations of the flare events made with a 15 cm solar tower telescope at ARIES, Nainital, India. The propagation characteristics of the CMEs have been derived from the three-dimensional images of the solar wind (i.e., density and speed) obtained from the interplanetary scintillation data, supplemented with other ground- and space-based measurements. The TRACE, SXI and H-alpha observations revealed two successive ejections (of speeds ~350 and ~100 km/s), originating from the same filament channel, which were associated with two high speed CMEs (~1223 and ~1660 km/s, respectively). These two ejections generated propagating fast shock waves (i.e., fast drifting type II radio bursts) in the corona. The interaction of these CMEs along the Sun-Earth line has led to the severity of the storm. According to our investigation, the interplanetary medium consisted of two merging magnetic clouds (MCs) that preserved their identity during their propagation. These magnetic clouds made the interplanetary magnetic field (IMF) southward for a long time, which reconnected with the geomagnetic field, resulting the super-storm (Dst_peak=-472 nT) on the Earth.Comment: 24 pages, 16 figures, Accepted for publication in Solar Physic

Проблема взаємозв’язку громадянського суспільства і державної бюрократії в Україні: деякі сучасні аспекти історіографії дослідження

Розглядається взаємодія громадянського суспільства і державної бюрократії. Зроблено висновок, що позиції українських дослідників відповідають напрацюванням західної наукової традиції стосовно теоретичних, а також практичних способів забезпечення взаємодії інститутів громадянського суспільства і державного апарату.In this article the interrelation between civil society and state bureaucracy is analysed. The conclusion is made that the views of the Ukrainian scientists correlate with the western traditional scientific opinion concerning theoretical and practical ways of ensuring interaction between the civil society institutions and state apparatus

Methodological considerations in the analysis of fecal glucocorticoid metabolites in tufted capuchins (Cebus apella)

Analysis of fecal glucocorticoid (GC) metabolites has recently become the standard method to monitor adrenocortical activity in primates noninvasively. However, given variation in the production, metabolism, and excretion of GCs across species and even between sexes, there are no standard methods that are universally applicable. In particular, it is important to validate assays intended to measure GC production, test extraction and storage procedures, and consider the time course of GC metabolite excretion relative to the production and circulation of the native hormones. This study examines these four methodological aspects of fecal GC metabolite analysis in tufted capuchins (Cebus apella). Specifically, we conducted an adrenocorticotrophic hormone (ACTH) challenge on one male and one female capuchin to test the validity of four GC enzyme immunoassays (EIAs) and document the time course characterizing GC me- tabolite excretion in this species. In addition, we compare a common field-friendly technique for extracting fecal GC metabolites to an established laboratory extraction methodology and test for effects of storing “field extracts” for up to 1 yr. Results suggest that a corticosterone EIA is most sensitive to changes in GC production, provides reliable measures when extracted according to the field method, and measures GC metabolites which remain highly stable after even 12 mo of storage. Further, the time course of GC metabolite excretion is shorter than that described yet for any primate taxa. These results provide guidelines for studies of GCs in tufted capuchins, and underscore the importance of validating methods for fecal hormone analysis for each species of interest