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Fish exhibit complex social behaviours that can influence their stress levels and well-being. However, little is known about the link between social interactions and stress in wild fish, especially in running water environments. While many studies have explored the stress axis in fish, most have focused on specific social contexts, leaving gaps in understanding stress responses to social changes. Our study investigated collective behaviour and stress in wild Italian riffle dace ( Telestes muticellus ) in a controlled experimental setup simulating a natural river system. Results reveal that group-living fish have lower cortisol and oxidative stress levels in muscle tissue compared to solitary counterparts, suggesting a calming effect of conspecific presence. Additionally, we observed upregulated expression of antioxidant enzymes in group-living fish, indicating potential benefits to antioxidant defence systems. These insights shed light on the dynamic relationship between group behaviour and stress in wild fish within running water habitats and emphasise the use of multidisciplinary approaches.

Heatwaves, increasingly prevalent in our rapidly changing climate, significantly impact animals with far‐reaching ecological and evolutionary consequences. One of the first responses in animals to stress, including heat stress, is behavioural change, and this can directly influence fitness and survival. Changes in anti‐predator behaviour are particularly critical, as they may compromise a prey's ability to evade predators, thus increasing predation risk and jeopardising survival. In the context of climate change, assessing anti‐predator reactions under ecologically relevant heat stress is thus crucial, especially during the vulnerable life stage of development. This study investigated the effects of a heatwave on anti‐predator responses in juvenile guppies (Poecilia reticulata). One‐month‐old guppies were subjected to a 5‐day experimental heatwave (32°C) or a control temperature (26°C). After the treatment, all individuals were tested at a common temperature (26°C) for anti‐predator behavioural responses and swimming performance, the latter serving as a proxy for physical condition. While heatwave exposure did not affect swimming performance, it significantly altered anti‐predator responses. Heatwave‐exposed juveniles exhibited a reduced freezing response and faster resumption of normal activity compared to control fish. Our findings demonstrate that heatwaves can modify prey's anti‐predator behaviours during critical developmental stages. This suggests that heatwaves may increase predation risk, potentially impacting survival rates and reshaping predator–prey interactions in the face of ongoing climate change. We assessed how a heatwave affects anti‐predator reactions of juvenile guppies (Poecilia reticulata), by tracking their activity in an open field setup. When presented with a predator cue, guppies that were previously exposed to a 5‐day experimental heatwave (32°C) displayed an attenuated freezing response and resumed normal swimming activity faster than guppies from a control temperature (26°C). The results show that heatwaves can change prey's anti‐predator behaviours, suggesting that heat stress induced by climate change may influence predation risk in the wild.

Microfluidic platforms have emerged as critical technologies for exploring sperm chemotaxis, providing precise gradient control, and facilitating in-depth behavioral assessment. We designed a novel, user-friendly microfluidic device that is optimized for human sperm morphology and motility. The device was validated using two well-established sperm chemoattractants, progesterone and bourgeonal, demonstrating its reliability and reproducibility. Given the key role of olfactory receptors (ORs) in mediating sperm chemotaxis, the newly developed device was employed to identify additional receptors that may contribute to sperm behavior. Using the Atlas database, we identified OR2H1 as a candidate receptor. It is enriched in testis-derived cells, particularly in early and late spermatids, and it is broadly expressed across human spermatozoa. We demonstrated that OR2H1’s ligand, methional, a sulfur-containing aldehyde naturally found in vaginal fluid and biosynthesized by Lactococcus lactis, significantly enhances sperm migration and progressive motility. Methional stimulation also triggered increased intracellular calcium levels, indicating receptor activation. Computer-assisted sperm analysis revealed that methional treatment improved sperm linearity, straightness, and wobble without affecting the average velocity, suggesting enhanced directional movement. These findings provide evidence that methional promotes sperm chemotaxis via OR2H1 and highlight the potential role of the vaginal microbiome in influencing human fertility.

There is considerable evidence that female reproductive fluid (FRF) interacts intimately with sperm, affecting several sperm traits, including sperm motility and longevity, and ultimately fertilization success. One of the first documented interactions between FRF and sperm is the ability of FRF to attract and guide sperm towards the eggs. However, most of the evidence of FRF’s chemoattraction proprieties comes from a limited number of taxa, specifically mammals and invertebrate broadcasting spawners. In other species, small FRF volumes and/or short sperm longevity often impose methodological difficulties resulting in this gap in chemoattraction studies in non-model species. One of the outcomes of sperm chemotaxis is sperm accumulation towards high chemoattractant concentrations, which can be easily quantified by measuring sperm concentration. Here, we tested sperm accumulation towards FRF in the zebrafish, Danio rerio, using an ad hoc developed, 3D printed, device (‘sperm selection chamber’). This easy-to-use tool allows to select and collect the sperm that swim towards a chemical gradient, and accumulate in a chemoattractant-filled well thus providing putative evidence for chemoattraction. We found that sperm accumulate in FRF in zebrafish. We also found that none of the sperm quality traits we measured (sperm swimming velocity and trajectory, sperm motility, and longevity) were correlated with this response. Together with the 3D printable project, we provide a detailed protocol for using the selection chamber. The chamber is optimized for the zebrafish, but it can be easily adapted for other species. Our device lays the foundation for a standardized way to measure sperm accumulation and in general chemoattraction, stimulating future research aimed at understanding the role and the mechanisms of sperm chemoattraction by FRF.

Ocean acidification poses an increasing concern for broadcast spawning species that release gametes in the water column where fertilization occurs. Indeed, the functionality of gametes and their interactions may be negatively affected by reduced pH. Susceptibility to other environmental stressors, such as pollutants, may be also altered under acidified conditions, resulting in more detrimental effects. To verify this hypothesis, combined exposures to CO2-driven acidification and environmentally relevant concentrations (0.5 µg/L) of three contaminants (caffeine, diclofenac, and PFOS, all singularly or in mixture) were carried out to highlight potential negative effects on fertilization success and motility of sperm in the sea urchin Paracentrotus lividus. Our results showed a significant reduction in the percentage of fertilized eggs when sperm were pre-exposed to reduced pH (ambient pH minus 0.4 units) compared to that of controls (ambient, pH = 8.1). Sperm speed and motility also decreased when sperm were activated and then exposed at reduced pH. Conversely, at both pH values tested, no significant effect due to the contaminants, nor of their interaction with pH, was found on any of the biological endpoints considered.

Objectives This study's primary purpose was to demonstrate the correlation of preoperative right ventricular free‐wall longitudinal strain (RVFWLS) and pre‐/postsurgical variation in strain (delta strain) with the clinical and echocardiographic diagnosis of right ventricular dysfunction. Its secondary purpose was to determine the correlation of RVFWLS and delta strain with length of stay (LOS) in the intensive care unit (ICU), ventilation days, trend of natriuretic peptide test. (NT‐proBNP) and lactate in the first 48 h, incidence of acute renal failure, and 28‐day mortality. Design Prospective observational study. Setting Cardio‐thoracic and Vascular Anaesthesia Department and ICU of the University Hospital Integrated Trust of Verona. Participants Patients scheduled for mitral surgery. Interventions None. Measurements and Main Results All clinical and transoesophageal echocardiographic (TEE) parameters were collected at baseline, before surgery (T1) and at admission in the ICU postsurgery (T2). During the postoperative period, the clinical and echocardiographic diagnoses of right, left, or biventricular dysfunction were evaluated. TEE parameters were evaluated by a cardiologist offline. The patients were divided into two subgroups according to the development of any type of ventricular dysfunction. No statistically significant differences emerged between the two groups. According to a logistic regression model, a T1‐RVFWLS value of −15% appeared to predict biventricular dysfunction (sensitivity: 100%; specificity: 91.3%). No correlation between T1‐ or T2‐RVFWLS and creatinine, hours of ventilation or ICU LOS was found. Conclusions Our study introduces a new parameter that could be used in perioperative evaluations to identify patients at risk of postoperative biventricular dysfunction.

Traits associated with mating and fertilization success are expected to senesce with age, but limited information is available on their relative rates of senescence. In polyandrous species, male reproductive fitness depends on both mating and fertilization success. Because successful mating is a prerequisite for post-copulatory sexual selection, ejaculate traits are expected to senesce faster than pre-copulatory traits, as pre-copulatory sexual selection is often deemed to be stronger than post-copulatory sexual selection. This pattern has generally been found in the few empirical studies conducted so far. We tested this prediction in the guppy ( Poecilia reticulata ), a livebearing fish characterized by intense sperm competition, by comparing the expression of male sexual traits at two ages (four and nine months). Contrary to prediction, we found that post-copulatory traits senesced at a significantly slower rate than pre-copulatory traits. We also looked at whether early investment in those sexual traits affects longevity, and the interaction between sperm age (duration of sperm storage inside the male) and male age. Our results suggest that the relative senescence rate of pre- and post-copulatory sexual traits may vary among species with different mating systems and ecology.

Female choice has been documented in many animal taxa, and how we test it has been refined through years of studies on the topic. However, when designing mate choice experiments some variables, surprisingly, often remain overlooked, including receptivity and reproductive stage. Here, we aimed to assess whether the female reproductive stage influences strength and direction of mate choice in the zebrafish, Danio rerio. Females were offered a choice between two males differing in body size. We found that female choice in our experimental setup was significantly repeatable and that females preferred larger males. Nonetheless, the level of choosiness of females was affected by the time since the last spawning. Females spent more time choosing when tested 7 and 10 days after spawning rather than 4 days, indicating a higher receptivity to males from one week after the last spawning. Moreover, females preferred larger males only when tested 7 and 10 days after spawning. Our results suggest that female mate choice should take female receptivity into account, by standardizing time since the last spawning across females. More broadly, this suggests that 7–10 days since the previous spawning is the ideal time interval for zebrafish female receptivity to peak, with implications for facilities and researchers to increase egg production in natural spawning events and manual egg collection.Significance statementThe role of pre-copulatory female mate preference has long been recognized in sexual selection. Nonetheless, female receptivity often remains overlooked in mate choice experiments especially in external fertilizing species. In the present study, we investigated if the female reproductive stage affects the strength and direction of female mate choice in an external fertilizing fish, the zebrafish, Danio rerio. We found that, when tested 7 and 10 days after spawning rather than 4 days, females spent more time choosing, demonstrating an increased receptivity to males from 1 week following the last spawning. Furthermore, only at 7 and 10 days after spawning females exibith a clear preference for the bigger males. Our study highlights the importance of considering the female receptivity in future studies assessing mate choices in this and other externally fertilizing species, and also for zebrafish facilities to increase egg production in natural spawning events and manual egg collection.

In many species, females mate with multiple partners, meaning that sexual selection on male traits operates across a spectrum that encompasses the competition for mates (that is, before mating) and fertilizations (after mating). Despite being inextricably linked, pre- and postcopulatory sexual selection are typically studied independently, and we know almost nothing about how sexual selection operates across this divide. Here we bridge this knowledge gap using the livebearing fish Poecilia reticulata . We show that both selective episodes, as well as their covariance, explain a significant component of variance in male reproductive fitness. Moreover, linear and nonlinear selection simultaneously act on pre- and postcopulatory traits, and interact to generate multiple phenotypes with similar fitness. In species in which females mate with multiple partners, sexual selection acts on male traits involved in mating and fertilization. Here, the authors show that selection acting before and after mating explains a significant component of variance in male reproductive fitness in a livebearing fish.

Traits associated with mating and fertilization success are expected to senesce with age, but limited information is available on their relative rates of senescence. In polyandrous species, male reproductive fitness depends on both mating and fertilization success. Because successful mating is a prerequisite for post-copulatory sexual selection, ejaculate traits are expected to senesce faster than pre-copulatory traits, as precopulatory sexual selection is often deemed to be stronger than post-copulatory sexual selection. This pattern has generally been found in the few empirical studies conducted so far. We tested this prediction in the guppy (Poecilia reticulata), a livebearing fish characterized by intense sperm competition, by comparing the expression of male sexual traits at two ages (four and nine months). Contrary to prediction, we found that post-copulatory traits senesced at a significantly slower rate than pre-copulatory traits. We also looked at whether early investment in those sexual traits affects longevity, and the interaction between sperm age (duration of sperm storage inside the male) and male age. Our results suggest that the relative senescence rate of pre- and post-copulatory sexual traits may vary among species with different mating systems and ecology.