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

Multi-Spacecraft Observations of the Evolution of Interplanetary Coronal Mass Ejections Between 0.3 and 2.2 AU: Conjunctions with the Juno Spacecraft

We present a catalogue of 35 interplanetary coronal mass ejections (ICMEs) observed by the Juno spacecraft and at least one other spacecraft during its cruise phase to Jupiter. We identify events observed by MESSENGER, Venus Express, Wind, and STEREO with magnetic features that can be matched unambiguously with those observed by Juno. A multi-spacecraft study of ICME properties between 0.3 and 2.2 AU is conducted: we firstly investigate the global expansion by tracking the variation in magnetic field strength with increasing heliocentric distance of individual ICME events, finding significant variability in magnetic field relationships for individual events in comparison with statistical trends. With the availability of plasma data at 1 AU, the local expansion at 1 AU can be compared with global expansion rates between 1 AU and Juno. Despite following expected trends, the local and global expansion rates are only weakly correlated. Finally, for those events with clearly identifiable magnetic flux ropes, we investigate the orientation of the flux rope axis as they propagate; we find that 64% of events displayed a decrease in inclination with increasing heliocentric distance, and 40% of events undergo a significant change in orientation as they propagate towards Juno. The multi-spacecraft catalogue produced in this study provides a valuable link between ICME observations in the inner heliosphere and beyond 1 AU, thereby improving our understanding of ICME evolution

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

Turbulence Properties of Interplanetary Coronal Mass Ejections in the Inner Heliosphere: Dependence on Proton Beta and Flux Rope Structure

Interplanetary coronal mass ejections (ICMEs) have low proton beta across a broad range of heliocentric distances and a magnetic flux rope structure at large scales, making them a unique environment for studying solar wind fluctuations. Power spectra of magnetic field fluctuations in 28 ICMEs observed between 0.25 and 0.95 au by Solar Orbiter and Parker Solar Probe have been examined. At large scales, the spectra were dominated by power contained in the flux ropes. Subtraction of the background flux rope fields reduced the mean spectral index from $-5/3$ to $-3/2$ at $kd_i \leq 10^{-3}$. Rope subtraction also revealed shorter correlation lengths in the magnetic field. The spectral index was typically near $-5/3$ in the inertial range at all radial distances regardless of rope subtraction, and steepened to values consistently below $-3$ with transition to kinetic scales. The high-frequency break point terminating the inertial range evolved approximately linearly with radial distance and was closer in scale to the proton inertial length than the proton gyroscale, as expected for plasma at low proton beta. Magnetic compressibility at inertial scales did not show any significant correlation with radial distance, in contrast to the solar wind generally. In ICMEs, the distinctive spectral properties at injection scales appear mostly determined by the global flux rope structure while transition-kinetic properties are more influenced by the low proton beta; the intervening inertial range appears independent of both ICME features, indicative of a system-independent scaling of the turbulence.Comment: 12 pages, 5 figures; accepted for publication in the Astrophysical Journal Letters 2023 September 2

Coronal Magnetic Structure of Earthbound CMEs and In Situ Comparison

Predicting the magnetic field within an Earth-directed coronal mass ejection (CME) well before its arrival at Earth is one of the most important issues in space weather research. In this article, we compare the intrinsic flux rope type, that is, the CME orientation and handedness during eruption, with the in situ flux rope type for 20 CME events that have been uniquely linked from Sun to Earth through heliospheric imaging. Our study shows that the intrinsic flux rope type can be estimated for CMEs originating from different source regions using a combination of indirect proxies. We find that only 20% of the events studied match strictly between the intrinsic and in situ flux rope types. The percentage rises to 55% when intermediate cases (where the orientation at the Sun and/or in situ is close to 45 degrees) are considered as a match. We also determine the change in the flux rope tilt angle between the Sun and Earth. For the majority of the cases, the rotation is several tens of degrees, while 35% of the events change by more than 90 degrees. While occasionally the intrinsic flux rope type is a good proxy for the magnetic structure impacting Earth, our study highlights the importance of capturing the CME evolution for space weather forecasting purposes. Moreover, we emphasize that determination of the intrinsic flux rope type is a crucial input for CME forecasting models. Plain Language Summary Coronal mass ejections (CMEs) are huge eruptions from the Sun that can cause myriad of space weather effects at Earth. The ability of a CME to drive a geomagnetic storm is given largely by how its magnetic field is configured. Predicting the magnetic structure well before CME arrival at Earth is one of the major goals in space weather forecasting. Palmerio et al. (2018) study 20 CMEs observed both at the Sun and at Earth. They use observations of the solar disc to determine the magnetic structure at the Sun and then compare it with the magnetic structure estimated via magnetic field measurements near Earth. They report that the magnetic structures match closely only in 20% of the events studied. They also estimate the orientations of the CME axes at the Sun and at Earth. They find that 65% of the events change their orientations by less than 90 degrees. They conclude that knowledge of the CME magnetic structure at the Sun is an important factor in space weather forecasting, but the CME evolution after eruption has to be taken into account in order to improve current predictions.Peer reviewe

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