Assessment of Biostimulation Methods based on Chemical Communication in Female Doe Reproduction

Biostimulation is an animal management practice that helps improve reproductive parameters by modulating animal sensory systems. Chemical signals, mostly known as pheromones, have a great potential in this regard. This study was conducted to determine the influence of short-term female rabbit exposure to different conditions, mainly pheromone-mediated, on reproductive parameters of inseminated does. Groups of 60 females/each were exposed to 1) female urine, 2) male urine, 3) seminal plasma and 4) female-female interaction, just before artificial insemination, and compared to isolated females controls (female-female separated). The following reproductive parameters were analyzed for each group: receptivity (vulvar color), fertility (calving rate), prolificacy and number of born alive and dead kits ⁄ litter. Our results showed that the biostimulation methods employed in this experiment did not significantly improve any of the analyzed parameters. However, female doe exposure to urine, especially to male urine, slightly increased fertility levels when compared to the rest of the experimental conditions. Female-female interaction before artificial insemination, which is a common practice in rabbit farms, did not have any effect, which suggests its removal to avoid unnecessary animal management and time cost. On the other hand, fertility ranges were lower for animals with pale vulvar color whereas no differences were noticed among the other three colours which measure receptivity (pink, red, purple), thus suggesting that these three colours could be grouped together. Additionally, equine chorionic gonadotropin injection could be replaced with various biostimulation methods, therefore reducing or replacing current hormonal treatments, and contributing to animal welfare and to a natural image of animal production.

A pheromone bouquet controls the reproductive behaviour of the male shore crab, Carcinus maenas

The reproduction of many brachyuran crustaceans involves the formation of mating pairs often around the time of the female moult with attraction of a sexual partner and mating behaviour controlled by sex pheromones. In shore crabs, Carcinus maenas, females produce sex pheromones that are released in the urine. High Performance Liquid Chromatography analysis (HPLC) of female urine shows that the pheromone, identified as the nucleotide uridine diphosphate (UDP), elutes as an unresolved peak with structurally related nucleotides. We examined female urine samples over the moult cycle and detected UDP as well as uridine triphosphate (UTP). Bioassays were conducted to establish the possibility of a blend of nucleotides forming a sex pheromone bouquet in C. maenas. Whilst UDP induced the male mate guarding behaviour (cradling), a mixture of the two nucleotides at a ratio of 4:1 UDP:UTP elicited an even stronger mating response than either UDP or UTP individually. The urine concentration and composition of these nucleotides changes over the moult period pre and post ecdysis, providing evidence that a pheromone bouquet composition is not always constant. The change of the bouquet is related to the physiological state of the sender, here the moult cycle. Our study unravels the functionality of reaction-specific molecules in a pheromone bouquet. Whilst UDP is the mating signal, UTP acts as an attractant and combined they maximise the reproductive response. The use of bouquets provides species-specificity, potentially enabling reproductive isolation of sympatric species, and contains valuable information on the physiological state of the sender.

How Women and Men Pee: Assessing Gender-Specific Urination Practices for a Comfortable Toilet Experience

NoMix toilets separate urine and feces at the source and are a promising resource recovery technology. However, design issues hamper the transformation from unattractive to aspirational products. Little effort has been done to design toilets that account for physiological differences, leading to adverse effects on user-friendliness and urine separation efficiency. We used infrared recordings to assess gender-specific urination practices. Based on field data, we developed the Urinator, a simple device that allows simulating male and female urine streams. This supports engineers in developing more user- and gender-friendly and reuse-oriented sanitation technologies.

Harnessing psilocybin: antidepressant-like behavioral and synaptic actions of psilocybin are independent of 5-HT2R activation in mice

Depression is a widespread and devastating mental illness and the search for rapid-acting antidepressants remains critical. There is now exciting evidence that the psychedelic compound psilocybin produces not only powerful alterations of consciousness, but also rapid and persistent antidepressant effects. How psilocybin exerts its therapeutic actions is not known, but it is widely presumed that these actions require altered consciousness, which is known to be dependent on serotonin 2A receptor (5-HT2AR) activation. This hypothesis has never been tested, however. We therefore asked whether psilocybin would exert antidepressant-like responses in mice and, if so, whether these responses required 5-HT2AR activation. Using chronically stressed male mice, we observed that a single injection of psilocybin reversed anhedonic responses assessed with the sucrose preference and female urine preference tests. The antianhedonic response to psilocybin was accompanied by a strengthening of excitatory synapses in the hippocampus—a characteristic of traditional and fast-acting antidepressants. Neither behavioral nor electrophysiological responses to psilocybin were prevented by pretreatment with the 5-HT2A/2C antagonist ketanserin, despite positive evidence of ketanserin’s efficacy. We conclude that psilocybin’s mechanism of antidepressant action can be studied in animal models and suggest that altered perception may not be required for its antidepressant effects. We further suggest that a 5-HT2AR–independent restoration of synaptic strength in cortico-mesolimbic reward circuits may contribute to its antidepressant action. The possibility of combining psychedelic compounds and a 5-HT2AR antagonist offers a potential means to increase their acceptance and clinical utility and should be studied in human depression.

The Role of Urine in Semiochemical Communication between Females and Males of Domestic Dog (Canis familiaris) during Estrus

This study aimed to assess the mechanisms of semiochemical signal detection in dogs. In the first experiment, five males were exposed to volatile semiochemicals emitted by a live female in estrus and the female’s urine sample collected during estrus. The odor of canine food and clean air were used as controls. In the second experiment, 25 males could directly sniff and lick the urine samples from females in estrus, from females in anestrus, from males and from humans, placed in a lineup. Sniffing, licking and salivation, as well as keeping dogs at different distances from the source of odor, were recorded in both experiments. Experiment 1 showed that food odor was sniffed by males longer than estrous urine. Volatile semiochemicals from females in estrus evoked interest in males but without visual cues did not cause overt symptoms of sexual arousal. In Experiment 2, the estrous urine evoked interest in males and provoked significantly longer sniffing. Licking accompanied by salivation was observed in all instances only during direct contact with estrous urine. The results suggest a complex character of detection of female reproductive status, in which both volatile and nonvolatile compounds emitted by females and present in female urine are involved.

Experimental Evaluation of Spontaneous Olfactory Discrimination in Two Nocturnal Primates (Microcebus murinus and M. lehilahytsara)

Solitary species often employ chemocommunication to facilitate mate localization. In the solitarily foraging, nocturnal mouse lemurs (Microcebus spp.), females advertise their short period of estrus acoustically and by increased scent marking, whereas males search widely for receptive females. Both sexes can be trained by operant conditioning to discriminate conspecific from heterospecific urine scent. However, it is not known, if males during and outside the reproductive season show different spontaneous interest in conspecific female urine, and if urine from estrous females elicits a higher investigation response than that from diestrous females. We established a spontaneous discrimination paradigm and quantified olfactory investigation responses of 21 captive male mouse lemurs of M. lehilahytsara and M. murinus when presenting 1 conspecific and 1 heterospecific female urine odor sample simultaneously. Overall, M. murinus investigated stimuli significantly longer than M. lehilahytsara. Moreover, males of M. murinus showed significantly longer olfactory investigation at conspecific urine samples during but not outside the reproductive season. This indicates that female urinary cues are spontaneously discriminated by male M. murinus and that this discrimination is more relevant during the reproductive season. However, males of both species did not show different responses toward urine samples from estrous versus diestrous females. Finally, male age did not correlate with the overall duration of olfactory investigation, and investigation levels were similar when testing with fresh or frozen urine samples. In conclusion, this new spontaneous discrimination paradigm provides a useful additional tool to study olfactory communication of nocturnal primates from the receiver’s perspective.

Female Urine-induced Ultrasonic Vocalizations in Male C57BL/6J Mice as a Proxy Indicator for Postoperative Pain

Objectively recognizing postoperative pain in mice is challenging, making it difficult to determine an appropriate postoperative analgesic regimen. Adult male mice produce ultrasonic vocalizations after exposure to adult female urine (FiUSV). To determine if FiUSV can be used as a indicator of postoperative pain, FiUSV produced by male C57BL/6J mice were assessed for 5 d before and after vasectomy or sham surgery with or without sustained-release buprenorphine. Postoperative pain was assessed by monitoring vocalization using an ultrasonic microphone and by evaluating orbital tightness, posture, and piloerection at postoperative time points. Before vasectomy or sham surgery, 25 of 38 male mice produced FiUSV on 4 of 5 d (143 ± 93 FiUSV). Vasectomized mice without postoperative analgesia produced significantly fewer FiUSV (59 ± 26 FiUSV) compared with baseline (212 ± 102 FiUSV) at 4 h postoperatively, but returned to baseline by 28 h. Vasectomized mice treated with buprenorphine and sham-surgery mice had no change in FiUSV from baseline at any time point after surgery. Activity was decreased compared with baseline in vasectomized mice, regardless of receiving postoperative analgesia or not, but only at the 4-h time point. There were no differences in behavior scores between vasectomized mice and sham-surgery mice at any time point. These results show that FiUSV can be used to detect postoperative pain in male C57BL/6J mice after vasectomy.

A Proposed Mating Guideline for Bornean Orangutans (Pongo pygmaeus) using Urinary Hormonal Tracking Methods

Bornean orangutans (Pongo pygmaeus) are a critically endangered species. Proper monitoring of the menstrual cycle is important in breeding programs. Hormonal cycle tracking was performed in this study by collecting female urine. Metabolites of estrogen (estrone glucuronide, E1g) progesterone (P4) were measured using enzyme immunoassay. E1g tracking was carried out for 8 animal-months during normal cycling to obtain an average hormonal profile. P4 was tracked throughout normal cycling to obtain a baseline and over 1 successful pregnancy to obtain a reference for pregnancy monitoring and parturition anticipation. This study concludes that this non-invasive hormonal cycle monitoring using urine is an acceptable method. A recommended mating guideline of introducing the male after an estrogen increase of 1.5 times the baseline is proposed based on the results. This minimizes coerced mating and increases chances of pregnancy, which would benefit animal welfare and greatly assist in the conservation of the species.

Female- and Intruder-induced Ultrasonic Vocalizations in C57BL/6J Mice as Proxy Indicators for Animal Wellbeing

Female urine-induced male mice ultrasonic vocalizations (FiUSV) are ultrasonic vocalizations produced by adult male mice after presentation of adult female urine, whereas intruder-induced ultrasonic vocalizations (IiUSV) are produced by resident adult female mice when interacting with an intruder female mouse. These affiliative behaviors may be reduced when mice have decreased wellbeing or are in pain and distress. To determine whether FiUSV and IiUSV can be used as proxy indicators of animal wellbeing, we assessed FiUSV produced by male C57BL/6J mice in response to female urine and IiUSV produced by female C57BL/6J mice in response to a female intruder at baseline and 1 and 3 h after administration of a sublethal dose of LPS (6 or 12.5 mg/kg IP) or an equal volume of saline. Behavior was assessed by evaluating orbital tightness, posture, and piloerection immediately after USV collection. We hypothesized that LPS-injected mice would have a decreased inclination to mate or to interact with same-sex conspecifics and therefore would produce fewer USV. At baseline, 32 of 33 male mice produced FiUSV (149 ± 127 USV in 2 min), whereas all 36 female mice produced IiUSV (370 ± 156 USV in 2 min). Saline-injected mice showed no change from baseline at the 1- and 3-h time points, whereas LPS-injected mice demonstrated significantly fewer USV than baseline, producing no USV at both 1 and 3 h. According to orbital tightness, posture, and piloerection, LPS-injected mice showed signs of poor wellbeing at 3 h but not 1 h. These findings indicate that FiUSV and IiUSV can be used as proxy indicators of animal wellbeing associated with acute inflammation in mice and can be detected before the onset of clinical signs.

Olfactory Laterality Is Valence-Dependent in Mice

(1) Background: Although olfaction is the predominant sensory modality in rodents, studies focusing on lateralisation of olfactory processing remain scarce, and they are limited to the exploration of brain asymmetries. This study aimed to test whether outbred and inbred mice (NMRI and C57BL/6J mice strains) show nostril-use preference in processing olfactory stimuli differing in terms of emotional valence under unrestrained conditions. (2) Methods: Five odour stimuli were used in the study: vanilla, female urine, garlic, rat, distilled water. We measured the number of times mice used their left or right nostril for each testing session. (3) Results: We here showed that mice preferentially used their right nostril when sniffing attractive stimuli (female urine, vanilla), and their left nostril when sniffing aversive stimuli (rat odour). Results were consistent for both strains. (4) Conclusions: Surprisingly, the results obtained seem opposite to the valence theory assessing that the left and the right hemispheres are dominant in processing stimuli with a positive and a negative valence, respectively. It remains to be determined whether this valence-dependent pattern is specific or not to olfaction in mice. These new findings will be important to better understand how both hemispheres contribute to odour processing in rodents.