Animals

Reproductive dysfunction remains a major challenge for Senegalese sole (Solea senegalensis) aquaculture. Hormone-induced ovulation and in vitro fertilisation (IVF) are currently used to overcome the absence of natural courtship behaviours in captivity. This study investigates the feasibility of hormone-free IVF and behaviour-based prediction of ovulation as alternative strategies to enhance reproductive outcomes. We selected males using computer-assisted sperm analysis to assess sperm motility and quality for IVF trials. IVF trials were conducted using selected males and naturally ovulated eggs collected from females during evening hours across six experimental nights in two groups. Fish behaviour was continuously recorded using underwater cameras, and a convolutional neural network was developed to automatically detect Rest the Head (RTH) and Locomotor Activity (LA) behaviours. These behavioural counts, together with timing information, were used as features to train a logistic regression model for predicting ovulation events. Hormone-free IVF achieved fertilization rates up to 44% with 18% hatching success, producing viable larvae without hormonal intervention. Both groups showed significantly elevated RTH and LA during ovulation nights compared to non-ovulation nights, with peak activity occurring between 18:00-19:00 hours. The behavioural prediction model correctly identified ovulation with 82-85% accuracy and an area under the curve of 0.95. These findings demonstrate that sperm-quality-based male selection combined with automated behaviour analysis provides a practical, non-invasive approach for hormone-free reproduction in organic flatfish aquaculture.

Most common housing systems for dairy cows restrict their movement, which can influence welfare, gait, and hoof health of dairy cows. Outdoor access has been proposed as a management practice to offset these restrictions, but reported effects on cows locomotion vary and may not always be captured by traditional clinical assessments. In this study, we investigated gait and hoof through clinical (i.e., visual locomotion scoring and hoof lesion assessment) and subclinical (3D motion analysis, kinetic assessment, hoof infrared thermography and measuring claw conformation) methods to assess how limited provision of outdoor access affects non-lame cows housed in movement restricted environment. Thirty-six Holstein tie-stall cows were either given 1day/week (EX1) or 3days/week (EX3) of outdoor access (1h/day) during 5 consecutive weeks. Clinical and subclinical assessments of gait and hoof were performed before (Pre-trial), after 5 weeks of outing (Post-trial) and 8 weeks after outing (Follow-up). The results of this study revealed no clinical effect of outdoor access on cows locomotion score and hoof lesion prevalence. However, for subclinical assessment, both groups showed an increase in stride and stance time at Post-trial, with an increase in pressure applied by cows while standing in EX3 group and a reduction in coronary band temperature in both groups at Post-trial and Follow-up. Contact area and claw conformation changed after provision of outdoor access in both groups. This study illustrates that with the use of subclinical methods; we can reveal effects of outdoor access on gait and hoof health that might not be visible using the traditional methods.

Cetacean pathology is a cornerstone for population and marine ecosystem health monitoring, allowing clear differentiation among natural and anthropogenic threats. Previous studies in the Canary Islands reported natural causes of death in 59.4% (1999-2005) and 81% (2006-2012) of stranded cetaceans, versus anthropogenic causes in 33.3% and 19%, respectively. This study aimed to determine the causes of death (CD), pathologic findings, and epidemiological patterns of 316 cetaceans stranded in the Canary Islands between 2013 and 2018. The CDs were classified in pathologic entities (PEs) emphasizing natural versus anthropic origins. Of 316 animals, 224 (70.9%) from 18 species were suitable for pathological investigations. Among natural PEE, natural pathology associated with good nutritional status (NP-GNS) and natural pathology associated with significant loss of nutritional status (NP-LNS) represented 43/224 (19.2%) and 36/224 (16%) cases, respectively. Natural pathology with undetermined nutritional status (NP-UNS) occurred in 19/224 (8.5%) animals. Intra- and interspecific traumatic interactions (ITI) represented 30/224 (13.4%) cases, followed by neonatal/perinatal pathology (NPN) 19/224 (8.5%) and live-stranding stress and/or capture myopathy (LS-CM) 18/224 (8%). Infectious and parasitic diseases predominated in natural PEs. Anthropogenic PEs included interaction with fishing activities (IFA) in 17/224 (7.6%) cases, vessel collisions (VC) in 9/22 (4%) cases, and foreign body-associated pathology (FBAP) in 3/224 (1.3%) animals. Overall, anthropogenic causes accounted for 12.9% of deaths, natural causes for 73.6%, and the CD could not be established in 30/194 (13.4%) cases. This study reaffirms the trends concerning recognized PEs (NP-GNS, NP-LNS, NP-UNS, ITI, NPN, LS-CM, IFA, VC, and FBAP), expands the body of knowledge on cetacean pathology in the Canary Islands, and reports novel findings including mixed infections, clostridiosis in uncommon species, uremic syndrome secondary to urethral nematodiasis, gas embolism in unusual species, epibiont stomatitis, congenital musculo-skeletal malformations, or neoplastic processes. These findings advance understanding of cetacean mortality patterns and support conservation and management strategies.

To investigate how the disturbances associated with a relocation to a bedded-pack barn, such as a housing system change, a milking system change and a social regrouping, impacts the behavior of lactating dairy cows, 38 cows from a total of 9 tie-stall or free-stall commercial farms were moved to a newly built bedded-pack barn on an enrollment basis, with a social regrouping occurring after 2 weeks. Scan sampling of video data was done to assess behavior expression in the pen, and live data was collected to assess milking reactivity and animal handling procedures. Results indicate that the cows adapted quickly to the relocation to the new housing system as there were no changes in the locations in the pen, the body positions or the behaviors of cows in time between arrival and regrouping. The social regrouping had a bigger impact with a decrease in 16% of the observed time spent lying and an increase of 9.7% of the observed time spent feeding. Cows also adapted quickly to the milking procedures with a rapid decrease in the occurrence of negative social interactions between cows at the parlor, and in needing less human-animal manipulations and less time to be brought to the parlor. The housing system of origin had a slight effect on behaviors with cows from tie-stalls spending 1.7 times more of the observed time lying than free-stall cows, and free-stall cows spending 1.6 times more of the observed time feeding than tie-stall cows. This study provides a better understanding of how dairy cows respond to disturbances and is encouraging for producers that need to make changes to their current housing system as cows were shown to be quickly adaptable to the challenges presented to them. SummaryDairy cows from cubicle systems were shown to adapt quickly after a relocation to a bedded pack barn, the first use of a milking parlor, and a social regrouping. This was supported by limited changes observed in their behaviors after the disturbances, and observed deviations were temporary and short-lived. Animal handling procedures also observed a quick improvement in time with the trips to the milking parlor needing 2x less time and 3.5x fewer physical contacts from handlers after 5 days. These results are encouraging to producers needing to make changes to their barns.

Introducing cognitive enrichment from an early age has the potential to enhance an animals capacity to learn both simple and complex tasks, promote neural plasticity, and support cognitive development. This is applicable for young cattle who are at a critical stage in their development and could benefit from the influence cognitive enrichment has on their behavioral expression. This study aims to explore the effects cognitive enrichment has on weaned dairy calves through analyzing behavioral measures of voluntary participation and short-term behavioral reactions to enrichment exposure. Our study involved a total of five pairs of weaned calves (n=8 treatment; n=2 control). The treatment groups were presented with three variations of a puzzle box, each equipped with unique challenges that offer different solutions (push, slide, pull). These boxes were provided to the calves twice daily over the span of nine days in an isolated corridor located behind their pen. We hypothesized that motivated calves would consistently engage with cognitive enrichment voluntarily over time and express directed natural behaviors, reflecting sustained participation across repeated trials. Results demonstrated that calves consistently visited the cognitive enrichment area across trials, with an average latency of 75.7 {+/-} 47.0s from the pen to the enrichment. Secondly, the calves spent a significant proportion of trial time within the enrichment area at 65% (870.1 {+/-} 21s). Lastly, all calves expressed a broad range of behaviors in line with their natural exploration within the enrichment area, while the puzzle box treatment groups expressed higher durations of behavioral expressions when compared to the control (F=11.7, p<0.0001). Combined, these results indicate the calves motivations to voluntarily participate in a cognitive challenge. While these are promising findings for cognitive enrichment and its applicability to dairy calves, further work is needed to understand broader parameters. Specifically, how can social dynamics influence enrichment interaction in groups, how can this type of enrichment be implemented on farms, and what are the long-term effects to providing cognitive enrichment in the early stages of development.

Restlessness is a symptom of chronic boredom in humans and a behavioural phenomenon anecdotally described as a concern in bulls raised for fattening purposes, but it has so far not been addressed in research. The two studies presented in this paper aimed to gain first insights into restlessness in bulls. We operationally defined restlessness by the number of transitions between behaviours in a given time period, and quantified restlessness in bulls of different weight classes (300, 400, 500 kg) on farms keeping bulls on fully-slatted floors (n=8, Study 1) as well as across three different husbandry systems (fully-slatted floor (FS, n=4), straw-based (SB, n=4) and organic pasture (OP, n=3), Study 2). All farms were visited twice, and the behaviour of different individuals was continuously recorded for 15 minutes each between 9 a.m. and 5 p.m. (Study 1) and for 8 minutes each between 6 a.m. and 10 p.m. (Study 2). The effects of weight class and husbandry system were analysed using generalised linear mixed-effects models, and we ran a sequence analysis to cluster observations by the sequence, frequency, and duration of bulls behaviours in Study 1. Bulls kept in fully-slatted floor systems in Study 1 changed their behaviour on average 48.3 times per 10 minutes, with high variability both within and across farms. Weight class did not have a statistically supported effect on the number of transitions, and the sequence analysis revealed four clusters that differed in sequence and in the number of transitions. In Study 2, OP bulls showed fewer transitions than SB and FS bulls (X22 = 23.6, p < 0.001), while SB and FS bulls did not differ. While SB pens were more structured and offered more space per animal, both SB and FS systems can be characterised by monotony, which may explain the similar level of restlessness in both systems. Alternatively, or in addition, the high feeding intensity in SB and FS systems may have caused the higher number of transitions compared to the OP system, potentially elicited by subacute ruminal acidosis and/or laminitis and the resulting pain. However, these explanations are speculative and require systematic disentanglement in future studies. This study provides initial insights into restlessness in bulls and lays the groundwork for future research to identify the causes underlying restlessness and investigate its association with bull welfare.

Animal welfare is a central aspect in animal-based research where mice are most commonly used. Their facial expression can be analyzed to assess their well-being status using the Mouse Grimace Scale. However, its manual application becomes increasingly impractical when used on a large number of animals. This lead to the ongoing integration of computer vision methods to automate the analysis. While such methods have proven effective qualitatively, a systematic assessment to verify their reliability largely remains an open research gap. In this work, we attempted to close this gap as we evaluated three dominant paradigms (i.e., classification from supervised learning features, self-supervised learning features, or landmark locations) for the binary (i.e., well-being un-/impaired) classification of facial mouse images. Our quantitative results showed that such methods can be employed successfully with as low as 16% type II error rates. For qualitative assessment, we visualized the decision-making process and demonstrated that mainly pixels associated with the mouse rather than its environment are used. We further discovered that visual characteristics of the mice beyond those described by the Mouse Grimace Scale contributed to the classification. Our work showed that the automated well-being status assessment in mice is trustworthy and urges towards widespread adoption.

As global reptile populations continue to decline, improving reproductive success in managed populations of listed species, such as Phrynosoma cornutum (the Texas horned lizard) has become increasingly critical for species survival. One understudied area of reproductive research in reptile species is gamete collection and storage, a crucial component for maintaining genetic diversity. In Texas, semen was collected from wild P. cornutum (n = 20) in June 2025. Semen collection was performed via electroejaculation (EEJ) under alfaxalone anesthesia. Prior to semen collection, snout-vent-lengths (SVL) and weights were recorded and testes measurements were taken using a portable ultrasound. Average sperm motility and concentration across all lizards was 83.7% and 85.7 x 106 sperm/mL, respectively. While lizards with longer SVLs had higher sperm motility, weight and testis size did not affect sperm parameters. Samples were extended in INRA96 and divided for use in cold-storage longevity or cryopreservation trials. Samples under cold-storage conditions were assessed for motility daily for 10 days. Motility was not significantly reduced until 48 hours post-collection and maintained 19% motility at day 10. For cryopreservation, samples were diluted 1:1 in INRAFreeze cryopreservation media and frozen in liquid nitrogen, then immediately thawed. Average post-thaw sperm motility was 13.9%, with the highest post-thaw motility recorded at 38.2%. This is the first report of semen storage and cryopreservation in Phrynosoma and provides valuable insight into semen storage potential in reptile species.

Monitoring and improving animal health and welfare is a pressing need. Technological advances are increasing our ability to do so in a range of settings but currently, there are limited uses in experimental settings for less common model animals. Red foxes (Vulpes vulpes) are maintained by the French National Agency for Food, Environmental, and Occupational Health and Safety (ANSES) for regulatory and research purposes, primarily related to its role as a European Union and French Reference Laboratory for rabies and Echinococcus spp. In 2022, four foxes were surgically fitted with internal implants that recorded body temperature and activity levels and subsequently inoculated with rabies virus (RABV). Retrospectively, we tested the potential of these two variables to provide an early warning for the onset of rabies symptoms. We applied two anomaly detection algorithms (Shewhart and EWMA) with varying confidence levels to the data sets and compared how early the different models could detect significant changes while limiting false alarms during the calibration period. We hypothesized that body temperatures would rise significantly and foxes would significantly alter their activity levels at the beginning of infection, both at an earlier stage than is detectable through direct observation. We found that foxes significantly changed their activity during infection. We were best able to detect these changes using the EWMA algorithm, in some cases producing consecutive alarms up to two weeks before the death of an animal, while limiting false alarms. We found no evidence of fever in any of the infected foxes and body temperature did not appear to be a reliable indicator of foxes health. While here we applied our methods to a particularly severe and rarely implemented model with a very small sample size, this proof of concept illustrates the potential of these methods for a wide range of other situations that would benefit from similar long-term monitoring, including experimental protocols with milder clinical signs and routine monitoring for unexpected declines in health or welfare.

AO_SCPLOWBSTRACTC_SCPLOWPasture access is widely recognized for its welfare, behavioural, and health benefits in horses compared to individual stabling. Our recent studies have shown that even short-term pasture exposure enhances welfare and induces lasting changes in gut microbiota, promoting health-associated bacterial populations. This study investigated the transcriptomic signature of 22 horses following a standardized pasture protocol, with peripheral blood gene expression analysed before pasture access (T0) and three months after returning to individual stabling (T1). First, using sPLS regression, we correlated gene expression profiles with behavioural welfare indicators measured at T0 and T1. Horses with pasture access exhibited significantly altered blood transcriptome compared to controls, with aggressiveness towards humans emerging as the strongest behavioural correlate of gene expression. Bioinformatic analyses revealed that aggressiveness-associated genes were linked to inflammation, apoptosis, and cell differentiation/growth. Then, horses with pasture access were divided in resilient and non-resilient according to the improvement of their behaviour after time on pasture. Differentially expressed genes post versus pre- pasture, compared between resilient and non-resilient horses for aggressiveness showed that inflammatory signals were downregulated in both subgroups, with a more pronounced effect in resilient horses. This suggests that resilient individuals are better equipped to modulate inflammatory responses in low-stress environments like pasture. Surprisingly, for unresponsiveness to the environment--a trait linked to depressive-like states--resilient horses displayed increased inflammatory signaling (e.g., IRF and CD40 activation) post-pasture, while non-resilient horses showed activation of anti-inflammatory PPAR signaling. Notably, non-resilient horses exhibited molecular signatures associated with organismal death, morbidity, and growth failure, indicating maladaptive physiological states. In contrast, resilient horses demonstrated activation of growth-related pathways (e.g., BMP2 and BMPR1A), suggesting a shift toward anabolic and developmental processes. This study underscores the behavioural and molecular benefits of pasture access for horses, particularly in reducing aggressiveness and inflammatory signaling in resilient individuals. The findings highlight the complex interplay between behaviour, inflammation, and resilience, with pasture access promoting adaptive physiological and molecular responses. Further research is needed to elucidate the long-term implications of these transcriptomic changes and their broader relevance to equine welfare.

The measurement of inbreeding has gained significance across diverse fields, including population and conservation genetics, agricultural genetics, breeding programs for animals and plants, and wildlife management. This is due to the fact that inbreeding leads to increased homozygosity and results in lower genetic diversity, rendering populations more vulnerable to environmental changes, diseases, and other stressors. High or mid-coverage whole genome sequencing (WGS) has been widely used for inbreeding estimation, but it is resource-intensive. We aimed to investigate the use of ultra low-coverage whole genome sequencing (ulcWGS) as a cost-effective alternative for inbreeding analysis. Domestic dogs were used for our study as their extensive breeding histories lead to populations with a wide range of inbreeding levels. We constructed a multi-breed reference panel from high-coverage WGS samples. Inbreeding in independent ulcWGS samples was then estimated using runs of homozygosity (RoH) and inbreeding coefficients (F). We modeled the relationship between these measures and sequencing depth using nonlinear regression, to generate inbreeding estimates relative to sequencing depth. Resulting relative RoH and F measurements were significantly correlated, with purebred dogs exhibiting more runs of homozygosity and higher inbreeding coefficients compared to mixed-breed dogs. Our findings demonstrate that ulcWGS can provide reliable and economical estimations of inbreeding, expanding accessibility to genetic monitoring.

Equine endometrosis is a major cause of subfertility in mares characterized by fibrotic remodeling of the endometrium. Although transforming growth factor beta 1 (TGF-{beta}1) is implicated in fibrogenesis, the relationship between endometrosis severity and transcripts associated with tissue maintenance and proliferation remains incompletely defined. Present study evaluated endometrial mRNA expression of IGF1, MKI67, TGFB1, and ACTA2 in relation to endometrosis severity and defined histopathological features. Forty-seven endometrial samples were graded according to the modified Kenney and Doig (KD) categories. Relative mRNA expression was quantified by RT-qPCR and histopathology was extended using a standardized feature-based assessment. TGFB1 mRNA expression was higher in category I+ than in categories I and III (p = 0.041) and in samples with glandular basal lamina disruption (p = 0.020). MKI67 mRNA expression was lower in samples with luminal epithelial erosion (p = 0.049). IGF1 mRNA expression correlated negatively with KD category ({rho} = -0.401, p = 0.015), glandular degeneration ({rho} = -0.340, p = 0.043), overall inflammatory infiltration ({rho} = -0.387, p = 0.020), lymphocytic infiltration ({rho} = -0.426, p = 0.010), and neutrophilic infiltration ({rho} = -0.448, p = 0.006). MKI67 correlated positively with ESR1 ({rho} = 0.887, p < 0.001). These findings indicate that early endometrosis-compatible lesions are associated with increased TGFB1 transcription and that epithelial damage is accompanied by reduced MKI67 expression. The inverse associations between IGF1 expression and both lesion severity and inflammatory infiltration support a link between progressive histopathological changes and reduced expression of a growth factor involved in tissue maintenance.

Effective management of Bovine Respiratory Disease Complex (BRDC) requires timely, non-invasive diagnostic tools to protect calf health and welfare. Among early clinical signs, coughing stands out as both frequent and informative. To explore its potential for early BRDC detection, we deployed an artificial intelligence (AI)-driven acoustic monitoring system that recorded over 2,730 hours of audio during a 30-day period. Four experimental pens, each housing seven calves and stratified by infection status and antibiotic treatment, were equipped with a dedicated microphone to enable targeted acoustic surveillance. This configuration enabled pen-specific detection of cough events, which were subsequently classified using an AI HuBERT-based model trained on 1,045 labelled clips. The classifier achieved 92% accuracy. Temporal patterns in cough frequency aligned with infection dynamics, treatment responses, and circadian patterns. Notably, AI-detected coughs consistently preceded clinical scores by 1-2 days, confirming the systems sensitivity to early respiratory disorders. These findings support the use of acoustic surveillance as a valid, scalable, and autonomous tool for continuous monitoring and early warning of respiratory diseases in calves. ImplicationsThis study demonstrates that AI-powered acoustic monitoring enables real-time, non-invasive detection of coughs in calves for early warning of respiratory diseases, outperforming traditional veterinarian clinical scoring by 1-2 days. Its high accuracy and sensitivity to respiratory infection dynamics and treatment effects position it as a scalable tool for precision livestock farming.

In wildlife forensic practice, species identification and estimation of the Minimum Number of Individuals (MNI) for highly processed specimens have long relied on weight-based conversion methods, which may result in underestimation of the number of individuals involved in a case. Focusing on confiscated casque products of the helmeted hornbill (Rhinoplax vigil), this study combines macroscopic morphological examination with mitochondrial DNA barcoding (16S rRNA, COI, and Cytb) to explore a more robust approach for individual quantification. The results demonstrate that the conventional "weight-based" approach overlooks critical biological information contained in anatomical structures and cannot accurately reflect the actual number of individuals involved. Based on this, we propose an anatomy-based criterion centered on the principle of structural uniqueness: specimens retaining biologically unique beak or casque structures should be directly assigned to a single individual, whereas weight-based estimation should only be applied when original anatomical features are entirely absent. In addition, considering material loss during processing, we propose approximately 85 g as a reference threshold for estimating the number of individuals in heavily processed solid casque products. This approach improves the scientific rigor and accuracy of forensic identification and provides reliable technical support for the conviction, sentencing, and law enforcement of wildlife trafficking cases involving helmeted hornbill and other endangered species.

Enterohemorrhagic Escherichia coli O157:H7 (EHEC) is an important zoonotic pathogen that disrupts intestinal epithelial barrier integrity and induces excessive inflammatory responses, thereby leading to impaired growth performance and intestinal injury. EHEC is also an important cause Hemolytic Uremic Syndrome (HUS) in children and older adults. In pig production, chitosan is considered a promising alternative to antibiotics due to its bioadhesive and antimicrobial properties, but the effects and underlying mechanisms of chitosan (COS) under pathogenic challenge remain to be elucidated. One hundred and eight pigs were randomly divided into three treatments: an unchallenged control group (CON), an EHEC-challenged control group (ECON), and an EHEC-challenged group supplemented with 100 mg/kg COS (ECOS). Results show that EHEC challenge increased the feed conversion ratio (FCR), increased inflammatory cytokine levels, disrupted intestinal morphology, and downregulated tight junction and nutrient transporter gene expression (P<0.05). Dietary COS supplementation significantly improved average daily gain (ADG) and FCR during day 6-14 (P<0.05). Moreover, COS reduced fecal shedding of total E. coli (P = 0.085) and EHEC, attenuated systemic inflammation by decreasing serum TNF- and IL-6 levels, and enhanced humoral immunity as indicated by increased IgA and IgM concentrations (P<0.05). Importantly, COS alleviated EHEC-induced intestinal injury by restoring villus height and villus-to-crypt ratio, with enhanced mucosal digestive enzyme activities, and upregulated expression of tight junction proteins (ZO-1 and occludin) and nutrient transporters (SGLT-1 and PEPT1) (P<0.05). In conclusion, these findings indicate that dietary COS improves growth performance in EHEC-challenged weaned pigs, with enhanced intestinal barrier integrity and nutrient transport capacity.

Area and time restrictions are widely used in fisheries management for their simplicity and conservation benefits. Static closures (STCs) often fail to protect migratory fish; therefore, real-time closures (RTCs) are increasingly being adopted. However, RTCs require extensive and rapid data collection and analysis. The vessel monitoring system and daily landing reports provide near real-time data on fishing activities and fish abundance. We conducted a management strategy evaluation of RTCs and STCs in the Gulf of Thailand short mackerel fishery to clarify the efficacy of using RTCs to minimise fishing mortality, while considering their appropriate use with surveillance data for a migratory fish. The results support RTCs as more flexible and requiring a smaller closure area compared with STCs to achieve management objectives. We recommend gathering CPUE data on a monthly basis and using the highest CPUE threshold level to define a closure unit; no unit should be shut down until all units achieve the threshold level. Our results validate the efficacy of the RTC strategy for curtailing fishing mortality of a mobile species and demonstrate the effective use of RTCs to mitigate uncertainty in the migratory patterns of the species in an otherwise unpredictable, fluctuating environment.

Minimizing handling of threatened and endangered fish has become increasingly important as populations have dwindled. To minimize handling in morphometric measurements, the HandsFreeFishing program has been developed for juvenile Chinook Salmon (Oncorhynchus tshawytscha). By segmenting a 2D image many morphometric measurements are able to be estimated; from these measurements a weight prediction model is built based on fish whose ground truthed weights were measured using a digital scale. While many segmentation methods may be used, here Metas Segment Anything model (SAM) is employed to produce segmentation masks of raw images. This model is open-source and easily used on any image (of any size) with good performance. In the proposed framework, the user supplies a bounding box around a target fish along with minimal orientation data (left or right facing, upside down or right-side up); the rest of the segmentation, feature extraction, and final weight prediction is completely automated. A main goal of the segmentation is to estimate the surface area of the side profile of the fish. Then, assuming an ellipsoidal shape, this surface area can be related to the volume of the fish, which is directly proportional to the weight. Even on a relatively small dataset of 149 images (fork length 27-90mm) our results confirm the predictive qualities of the morphometric features measured. The model achieved weight prediction with a mean absolute error of 0.16 g with a mean absolute percentage error of 12%, and an r-squared value of 0.99, on fish ranging from 0.31g - 7.74g. The raw images come from a variety of fish viewers, the design of which is relatively inexpensive and reproducible, and, in conjunction with the HandsFreeFishing program, allows for minimal handling compared to traditional length and weight measurement methods.

O_LIStudying the reproduction process, which is a key determinant of individual and population fitness in endangered species, is challenging but urgently needed. The crested ibis (Nipponia nippon), a flagship endangered species recovering from an extreme population bottleneck, provides a valuable opportunity to examine how life-history strategies shape reproductive success and inform future conservation practices. C_LIO_LIWe monitored 176 breeding pairs of crested ibis over three consecutive breeding seasons and investigated the effects of three key life-history traits, namely breeding timing, clutch size, and nesting strategy (solitary versus colonial), on reproductive success (hatching and fledging success). C_LIO_LIOur analysis found that both hatching and fledging success declined significantly as breeding initiated later, and a positive association between clutch size and reproductive success in this species. These patterns were robust and repeatable across three years. Unlike other closely related species in this family, sibling competition is generally non-lethal, leading to large clutch sizes fledged in this endangered species. We consider this pattern to be a main reason underlying the rapid population recovery observed in the crested ibis. On the other hand, nesting strategy (colonial vs. solitary breeding) had no detectable effect on reproductive success. This pattern indicates the crested ibis can adopt different breeding strategies across habitats, highlighting its capacity to flexibly adjust breeding behavior in response to local environmental conditions. C_LIO_LIOur results provide an integrative assessment of how key life-history traits shape reproductive outcomes in a wild population of the crested ibis, serving as a foundation for evaluating its current status of population recovery and refining future conservation strategies for endangered avian species sharing similar life-history characteristics. C_LI

Habitat fragmentation, climate change, poaching, human-wildlife conflicts, and infectious diseases are the main threats to biodiversity conservation. They alter host-pathogen dynamics, reduce viable conservation areas, and promote genetic isolation, resulting in physiological stress among animal populations. Moreover, increased proximity between domestic and wild animals further facilitates disease spillovers exposing naive host species and ecosystems to new pathogens. Of the more than 200 known zoonotic diseases, approximately 60% originate from animals, contributing significantly to the global infectious disease burden. Here, we describe the development of an innovative non-invasive approach for biological sampling that has been validated in mice and shelter cats. Our device consists of a disposable plastic cassette that through odor attractants lures animals to lick a filter paper. This saliva collection approach allowed for the detection of RNA viruses by RTqPCR and third-generation sequencing. RTqPCR oral swab and licked paper results showed that both methods significantly predicted serological status. Our sequencing results revealed the richness of the gene space, demonstrating the potential of this device for discovering rare or unknown species circulating in the saliva donor, enabling this player to be recognized as an environmental sentinel. This study demonstrates the feasibility of deploying this device in sheltered/captive animal settings as well as under laboratory simulations of different environments, providing necessary foundations for future field applications. Our methodology holds great potential for monitoring zoonotic pathogens in both captive and free-ranging animals, to even possibly allow proactive mitigation measures prior to spillover, without interfering with the natural animal behaviour and social structures. Visual abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=141 SRC="FIGDIR/small/705368v1_ufig1.gif" ALT="Figure 1"> View larger version (21K): org.highwire.dtl.DTLVardef@1325498org.highwire.dtl.DTLVardef@14ff0cdorg.highwire.dtl.DTLVardef@14a4334org.highwire.dtl.DTLVardef@8217e4_HPS_FORMAT_FIGEXP M_FIG C_FIG

From facial expressions to gestures, animals use multiple signal modalities to express emotions and communicate. In dogs, tail movements are conspicuous behaviours associated with emotional states, but this link remains debated. We investigated canine emotional states underlying tail wagging by systematically analysing differences in tail movements in a computer-controlled task encompassing two non-social Conditions - Rewarded (positive) and Unrewarded (negative), and two Epochs (pre-response and outcome anticipation). Using pose-tracking we found that 11 out of 23 dogs did not wag their tails in at least 75% of trials, suggesting that some dogs may inherently wag less or that tail wagging is primarily a social signal. Our results showed that dogs were more likely to wag during positive anticipation; whereas in the negative condition, despite tail amplitude being more prominent, increased speeds reflected arousal rather than valence. Further work should assess tail kinematics in social contexts to test and extend these findings.