Breeding Season Research Articles

A case study of the 2023 highly pathogenic avian influenza (HPAI) outbreak in tern (Sternidae) colonies on the east coast of the Republic of Ireland

ABSTRACT Capsule:HPAI H5N1 had a significant impact on breeding Common Terns Sterna hirundo, Arctic Terns Sterna paradisaea and Roseate Terns Sterna dougallii on the east coast of the Republic of Ireland (ROI) in 2023, with a lesser impact on Sandwich Terns Thalasseus sandvicensis and only two cases among Little Terns Sternula albifrons. Aims: To document the outbreak of HPAI H5N1 in the tern colonies on the east coast of ROI during the 2023 breeding season. Methods:Precautions were taken from the start of the breeding season, including personal protective equipment (PPE) and safe operating procedures. Regular meetings were held between BirdWatch Ireland and the National Parks and Wildlife Service (NPWS) to respond to the emerging situation. Once HPAI was confirmed in a colony, all ringing and nest visits were stopped. Carcass removal was carried out at all affected colonies and most carcasses were incinerated. Results: Common Terns suffered the highest mortality across all colonies in 2023, with smaller but notable numbers of dead Roseate Terns and Arctic Terns, but very few Sandwich Terns and only two Little Tern deaths were attributed to HPAI. Conclusion: Regular meetings between relevant organizations were invaluable in preparing for outbreaks and ensuring an adaptive approach as the situation changed over time and between colonies. There was no way of knowing if the actions taken were effective, as the pattern of virus spread between species and sites remains poorly understood. Future clarity over the role of various state agencies and other relevant bodies in responding to HPAI in wild birds, greater sharing of real-time information, and pre-arranged agreements to ensure rapid testing and disposal of carcasses, would be useful to ensure an effective response to future outbreaks.

Prevalence of immunoglobulin E sensitisation to mites and pollens in 25,451 French dogs from 2016 to 2022.

Canine atopic dermatitis (cAD) is a common skin disorder in dogs, wherein allergens are presumed to exert a significant influence. The prevalence of allergen sensitisation in dogs with suspected cAD remains inadequately investigated in France. This study aimed to analyse allergen-specific immunoglobulin (Ig)E test results from French dogs with suspected cAD, exploring potential risk factors influencing sensitisation. The study included 25,451 client-owned dogs presented with clinical signs of cAD between January 2016 and December 2022. The study population consisted of 226 pure breeds and 915 cross-bred dogs of both sexes, with a mean age of 4.4 (±2.8) years. Data collection involved the compilation of results from canine serological allergen-specific IgE ELISA tests, using a 23 allergen composite panel. The influence of breed, sex, age, geographical area and sampling season on IgE test outcomes subsequently was analysed. Of the 25,451 analysed samples, 75.6% exhibited significant serum IgE concentrations in response to at least one of the tested allergens, with 67.9% testing positive for mites and 39.8% for pollens. Breed, sex, age, geographical area and sampling season were identified as significant risk factors influencing sensitisation patterns. This study provides insights into the prevalence and risk factors associated with allergen sensitisation in dogs with suspected cAD. Understanding these factors is crucial for effective diagnosis and tailored management strategies in veterinary practice. The findings underscore the need for personalised approaches in addressing cAD, considering individual patient characteristics.

House sparrows do not show a diel rhythm in double-strand DNA damage in erythrocytes.

DNA damage can be caused by a number of intrinsic and extrinsic factors. A recent study showed that free-living house sparrows (Passer domesticus) have higher DNA damage in the summer than the winter across five different tissues. This result was consistent when house sparrows were brought into captivity and exposed to comparable light cycles, with all other variables held constant. These results generated two hypotheses: (1) seasonal variation in DNA damage is related to circadian regulation and (2) seasonal variation in DNA damage is related to the total number of active hours. To investigate these hypotheses, we first quantified erythrocyte DNA damage in wild-caught house sparrows held in captivity on a 12L:12D light cycle at six points during the day to assess a diel or circadian rhythm but did not find one. We then performed a resonance experiment, in which birds experienced unnatural light cycles, and compared DNA damage in birds held on 6L:6D and 4.5L:7.5D resonance light cycles with their natural counterparts, 12L:12D and 9L:15D, respectively. We assessed corticosterone levels and DNA damage in blood before and after the resonance light cycles and DNA damage in abdominal fat, hippocampus, hypothalamus, and liver after the resonance light cycles. While our second experiment was not able to effectively test our hypotheses, we were able to demonstrate some interesting patterns. Throughout the resonance experiment, baseline corticosterone and testes size increased, consistent with the birds being photostimulated and preparing to breed. Surprisingly, the direction of change of DNA damage throughout the resonance photoperiod differed with tissue, which is not consistent with patterns during the breeding season in the wild. Our data indicate a potential uncoupling of the breeding physiology with the effect on DNA damage due to exposure to a resonance light cycle, which the birds may have interpreted as a skeleton photoperiod. Finally, though we were unable to fully disentangle the dynamics underlying seasonal DNA damage, we show that the previously documented patterns are not simply due to diel changes or the total amount of light exposure within a 24-hour period.

Captive Breeding Reveals Insights Into the Ecology and Reproductive Biology of 11 Little-Known Malagasy Frog Species.

Amphibians are facing an extinction crisis, with ex situ programs increasingly being used as a tool for their conservation. However, conservation efforts are often limited because we do not understand the ecological, behavioral, and life history traits of many amphibian species. Here, we report on the seasonal breeding patterns, egg-laying behavior, clutch size, and development of 11 frog species maintained at a conservation breeding facility in Andasibe, Madagascar. The frogs exhibited diverse breeding strategies aligned with life history theory. Counting the eggs in 1239 egg masses across these 11 species, we found endotrophic microhylids and terrestrial-breeding species had the smallest clutch size yet completed metamorphosis quickly, whereas species that laid eggs above or in water with exotrophic larvae had larger clutch sizes and took longer to develop. Most reproduction in captivity occurred during the warm, rainy season and followed seasonal patterns in temperature variation. Yet, Mantidactylus betsileanus bred throughout the year, and Heterixalus betsileo required additional environmental stimuli to trigger reproduction. Notably, we confirmed that Gephyromantis mitsinjo lays eggs on land with tadpoles developing terrestrially within jelly, a behavior previously theorized but which until now remained unobserved. Such observations show how captive breeding programs can be used to gain valuable data on the life history traits of species that are otherwise challenging to observe in nature. Our findings can be used to assess threats to closely related species, helping inform conservation efforts in a country harboring exceptional amphibian species richness and endemism.

Large invasive herbaceous plants decrease the taxonomic, functional and phylogenetic diversity of birds via their reproductive traits

Abstract Birds are among the organisms most impacted by plant invasions, the effects of which are particularly conspicuous during the breeding season. Birds can incorrectly assess the suitability of large, invasive hogweeds Heracleum sp. in south‐eastern Poland when selecting nesting sites early in spring when the developing invaders do not yet differ substantially in form from the rest of the vegetation. One would expect lower bird breeding success in areas with invasive hogweeds. Furthermore, this may shift bird community composition towards species with filtered traits, which may then be reflected in various measures of diversity. To assess these expectations, we conducted bird surveys at 74 sites, arranged as 37 pairs (with invasive Heracleum vs. control). The presence of Heracleum was associated with lower bird taxonomic and phylogenetic diversity. Functional richness (calculated based on species' reproductive traits) was also lower at sites with these invaders. The species detected at Heracleum sites were characterised by traits associated with rapid breeding, for example, small clutches, short incubation periods and short fledging periods. We show that the bird community at sites with hogweeds became less diverse and composed of random species sharing similar reproductive traits. Bird communities with low diversity at Heracleum sites tended to exhibit a common trait syndrome. These patterns provide insights into the mechanisms of how invasive plants may lead to the loss of some traits and species in bird communities. Read the free Plain Language Summary for this article on the Journal blog.

Limited evidence for the influence of the physical structure and floristics of habitat on a boreal forest bird community

Studies conducted at regional and continental scales show that avian richness, community composition, and abundance are associated with variation in the vegetation physical structure and community composition (floristics) within broad habitat types. The relative contributions of physical structure and floristics are contested, and relationships are often taxon-specific. We used 110 microphone arrays deployed across 90 sampling locations and two breeding seasons to survey an avian community in the boreal forest in Labrador, Canada. Our objectives were (1) to describe the avian community of an underrepresented portion of the boreal forest, (2) to estimate the relationships between avian species richness and habitat characteristics, (3) to estimate if species detection at a given location was related to local habitat characteristics, and (4) to investigate the spatial and temporal patterns of the avian community composition. We detected 32 species at our sampling locations; physical structure and floristics were not related to avian species richness, although estimates of richness were higher on warmer days and lower on windier days. Habitat characteristics were associated with the detection of Boreal Chickadee (Poecile hudsonicus), Yellow-rumped Warbler (Setophaga coronata), American Robin (Turdus migratorius), and Black-throated Green Warbler (Setophaga virens). Finally, avian community composition was only moderately consistent among three categorical forest types and between audio sampling periods in two consecutive breeding seasons. Overall, we show that the structural and floristic traits measured at our study site are not related to the detection of most avian species or to species richness.

Bird deterrent measures at a landfill decreased the productivity of a dependent population of White Stork (Ciconia ciconia)

Landfills provide an abundant and predictable food source for avifauna. The energy and time that are saved because of landfill‐foraging have had positive effects on the distribution of bird breeding populations and their reproductive parameters. However, the proliferation of individuals coinciding with the appearance of landfills often increases human–wildlife conflicts and intensifies the contact between waste and the environment. In this context, a landfill in Madrid (Spain) implemented deterrent measures in 2021 aiming to reduce the influx of birds inside its facilities. This study aims to describe the effects that a reduction in the accessibility and availability of landfill food resources may have had on the surrounding breeding populations of White Stork Ciconia ciconia. For this purpose, the breeding parameters of three populations with different landfill use indices were analysed before and after the application of bird deterrent measures. The closest population, with the highest landfill use index, suffered a drastic reduction in fledgling productivity during the breeding season with bird deterrent measures. On the other hand, a drought during one of the breeding seasons negatively affected the productivity of the populations that relied partially and completely on natural food resources (located at medium and long distance from the landfill, respectively). Landfill‐foraging might have mitigated the consequences of the natural food scarcity caused by this drought, even with the application of bird deterrent measures. Overall, our results show the potential negative impact of bird deterrent measures on populations dependent on landfill food, and highlight the importance of assessing the effect of these techniques beyond the site to determine their appropriateness. Due to the imminent closure of landfills and the expected worsening consequences of climate change, monitoring programmes should be established to determine the long‐term effects of bird deterrent measures and unusual environmental conditions on White Stork populations.

Avoidance and attraction responses of kittiwakes to three offshore wind farms in the North Sea

AbstractSeabird collision risk is a key concern in relation to the environmental impacts associated with offshore wind farms (OWFs). Understanding how species respond both to the wind farm itself, and individual turbines within the wind farm, is key to enabling better quantification and management of collision risk. Collision risk is of particular concern for the black-legged kittiwake, Rissa tridactyla, where modelling predicts unsustainable population level impacts. In this study 20 adult breeding kittiwakes, were tracked with GPS from Whinnyfold, Scotland (57°23′07″N, 001°52′11″W) during the breeding season in 2021. An Avoidance-Attraction Index (AAI) was estimated at several bands within macro- and meso-scales (0–4 km from outer boundary and 0–400 m from turbines, respectively), and the Avoidance Rate (AR; used in environmental impact assessments) at macro-scale to estimate avoidance behaviour to three operational OWFs within their foraging range. One offshore wind farm and its buffer zone (0–4 km from outer boundary) was visited more frequently by the majority of tracked individuals (19/20 birds), despite being twice as far as the closest OWF (17.3 and 31.9 km respectively), whilst 10 or less individuals used the remaining two OWFs. At the most frequented OWF we found macro-scale attraction to the closest band (0–1 km) trending towards avoidance in the furthest band (3–4 km). At the meso-scale we found avoidance of areas below the rotor height range (RHR, a.k.a. rotor swept area/zone) up to 120 m from individual turbines, which decreased to 60 m when within the RHR. Our results indicate that kittiwakes may be slightly attracted to the area around OWFs or aggregate here due to displacement but avoid individual turbines. Increased productivity in the OWF area may potentially be drawing birds into the general area, with aversion to individual turbines being responsible for meso-scale observations.

Drivers of nest site selection and breeding success in an Alpine ground-nesting songbird

AbstractBirds breeding in high-Alpine habitats must select a suitable breeding site and achieve successful reproduction within a restricted time. During four breeding seasons, we monitored nest sites of the Northern Wheatear (Oenanthe oenanthe), a high-Alpine long-distance migrant. We investigated how ecological factors predicted the selection of a site for nesting within the home range, using conditional logistic regression. Birds preferred south-exposed productive pastures on gentle slopes, interspersed with non-vegetated ground and human-made rockpiles. The direct vicinity of conspecific nests was avoided, as were shrubby or north-exposed areas. We investigated if habitat also influenced breeding success. We analysed the impact of environmental factors on breeding success, which was primarily driven by predation. The probability of the brood fledging successfully decreased on north-exposed slopes or on areas with low coverage of non-vegetated ground. The vicinity of conspecific nests did not have a clear effect. Further, we describe how breeding success varied within and between years. Within years, replacement broods had a higher breeding success. The apparent absence of variation in breeding success between years and a delay of the breeding period in the year with late spring onset suggest a high level of tolerance with respect to inter-annual variation of meteorological conditions. Since the preferred habitat is still widely available in the Alps and given the negative population trends in Western Europe, the Alpine range might serve as a refuge for the Northern Wheatear, as long as low-intensity management and heterogenous habitats are maintained.

Adaptive divergence in reproductive seasonality and underlying physiological features fit Rattus norvegicus to live as opportunistic breeders.

How organisms respond to complex environments is one of the unsolved problems in ecology. Life history patterns of a species provide essential information on how different populations may respond and adapt to environmental changes. Compared to typical seasonal breeders, which have limited distributions, the worldwide distribution of brown rats (Rattus norvegicus) across highly complex and divergent habitats suggests they exhibit exceptional adaptiveness. However, the difference in physiological mechanisms by which brown rats respond and adapt to markedly different environments is seldom investigated. Here, we reveal a significant divergence in reproductive seasonality and environmental responses between two brown rat subspecies: one subspecies, R. n. caraco, lives in the temperate zone, and another subspecies, R. n. norvegicus, lives in the subtropical region. Although R. n. caraco displayed a significantly higher reproductive seasonality than R. n. norvegicus, both subspecies adapted to sub-optimal breeding conditions mainly by regulating the seminal vesicle rather than testis development. Especially in responding to severe winter conditions in high-latitude regions, bodyweight-dependent recovery of testicular development in adults enables R. n. caraco to initiate reproduction more rapidly when conditions are suited. These findings elucidate a regulatory process of how brown rats live as opportunistic breeders by benefiting from enhanced semen production.

Passing plastic: Traces of plastic in the fecal samples of a high Arctic seabird in Tunu (East Greenland)

Arctic seabirds are key bio-indicators of marine plastic pollution due to their transient movement and large populations. Although many studies have quantified the ingestion of microplastic particles (<5 mm in size) through necropsy or regurgitation sampling methods, little is known about post-digestive particle excretion. Due to the logistical challenges of non-lethally sampling feces from Arctic seabirds, this pathway remains largely understudied. We non-lethally collected 110 fecal samples from little auks (Alle alle) during the 2014 breeding season in Ukaleqarteq, Tunu (Kap Höegh, East Greenland). We identified 25 potential microplastic particles (>100 µm in size), 19 of which were analyzed for material composition using Fourier Transform Infrared Spectroscopy and Raman micro-spectroscopy. Of these, 13 particles were successfully matched to materials, with five as plastic. This produced an average concentration of 0.08 ± 0.28 microplastic particles per fecal sample, with no difference of occurrence between chicks and adults. Particle lengths ranged from 113 - 751 µm. The presence of microplastics larger than our lower limit of detectability of 100-µm suggests a need for analysis of smaller particles and microfibers in this species. We contribute to understanding how microplastics pass through little auks and characterize how this species interacts with the plastic pathways.

Beyond tracking: Advancing animal ecology through the longitudinal sampling of individuals

Abstract Rarely, is there an opportunity to study and directly observe the same migratory animal across both breeding and nonbreeding seasons. In a rare, documented case, we highlight a live reencounter of a female Setophaga ruticilla (American Redstart) on its nest during the breeding season—an individual that was originally captured and tagged on its nonbreeding territory in Jamaica more than 3,250 km away. Advances in tracking technology allow researchers to track the movements of smaller organisms at higher resolutions. While tracking data alone have provided critical advances in the ecology of migratory animals, especially small migratory birds, there are exciting opportunities to move beyond tracking and to deeply explore biological questions about migratory animals by locating and directly studying (e.g., direct observation and physical sampling) the same individuals across the year. Although we have had the ability to precisely track large-bodied migratory birds (>100 g) for decades, researchers have rarely used these data to localize and directly observe or sample them across the year. Ultimately, we must consider the potential advances to be made in avian ecology through this endeavor. To this end, we highlight several specific areas of research that we believe move beyond tracking and can only be addressed by integrating direct longitudinal studies of individuals.

Oxidative state is associated with migration distance, but not traits linked to flight energetics

Flight can be highly energy demanding, but its efficiency depends largely on flight style, wing shape and wing loading, and a range of morphological and lifestyle adaptations that can modify the cost of sustained flight. Such behavioural and morphological adaptations can also influence the physiological costs associated with migration. For instance, during intense flight and catabolism of reserves, lipid damage induced by pro‐oxidants increases, and to keep oxidative physiological homeostasis under control, the antioxidant machinery can be upregulated. Studies on the oxidative physiology of endurance flight have produced contradictory results, making generalization difficult, especially because multispecies studies are missing. Therefore, to explore the oxidative cost of flight and migration, we used samples collected during the breeding season from 113 European bird species and explored the associations of measures of antioxidant capacity (total antioxidant status, uric acid and glutathione concentration) and oxidative damage of lipids (malondialdehyde) with variables reflecting flight energetics (year‐round or specifically during migration) using a phylogenetic framework. We found that none of the traits predicting year‐round flight energy expenditure (flight style, wing morphology and flight muscle morphology) explained any measures of oxidative state. Our results suggest that birds endure their everyday flight exercise without or with low oxidative cost. However, oxidative damage to lipids and one component of the endogenous antioxidant system (uric acid), measured after the end of spring migration on breeding adult birds, increased with migration distance. Our results suggest that migration could have oxidative consequences that might be carried over to subsequent life‐history stages (breeding).

Factors determining semen sample collection and semen quality parameters in African penguins Spheniscus demersus

Our research focuses on semen collection from 42 African penguin males, grouped by age, habituation levels, and reproductive season timing. We assess behavioral and physiological responses to dorso-abdominal massage, evaluating species-specific seminal traits using conventional and advanced methods. Positive behavioral responses corresponded with successful semen collection. Young and human-reared males exhibited more favorable behaviors, and samples containing spermatozoa were more likely collected during the reproductive season. Age did not influence sample collectability or spermatozoa presence, though mature males produced higher semen volumes. Young males exhibited more sperm morphological abnormalities, especially outside the breeding season. Sperm concentration and number per ejaculate showed no significant differences across age or seasonal groups. Young males had higher motile and progressive sperm percentages, while mature males had more static sperm. Additionally, percentages of live sperm and intact acrosomes were higher in mature males. Young males and samples from non-reproductive season presented more dead sperm with damaged acrosomes. Plasma membrane integrity positively correlated with age for live sperm and negatively for dying sperm. This research demonstrates the feasibility of semen collection from African penguins regardless of habituation level, fills the knowledge gap by describing sperm morphological abnormalities, and provides insights into using flow cytometry in Sphenisciformes.

Boldness as an Individual Trait of Behaviour in a Natural Population of Birds

Individual resoluteness during the resumption of feeding of nestlings was examined in tests involving the presentation of an unfamiliar object near the nests of the pied flycatcher (Ficedula hypoleuca) in a natural environment. Over the course of five breeding seasons, 229 individuals were tested, 41 of which were tested 2 to 4 times. Repeat testing of the same birds was conducted at different nests with intervals ranging from 3 to 1103 days (Med=367). We took into account the number and age of nestlings, nesting periods, the sex and age of the individual and its partner, the presence of molting, the intensity of nestling feeding, the behavior of the partner, as well as the presence of simultaneous bigamy in the male. We revealed a significant repeatability of test results over substantial time intervals (R=0.21), indicating the individual’s stability over time in response (boldness) to changing conditions in the natural environment.

Characterization and phylogenetic analysis of vomeronasal receptors in the female muskrat (Ondatra zibethicus)

Vomeronasal receptors (VRs) play a crucial role in recognizing pheromones, which are essential for social chemical communication. The male muskrat (Ondatra zibethicus) secretes musk, which contains pheromones as a reproductive signal, and the female can recognize it through the VNO to mediate social communication behavior. This study aimed to identify the genomic information of VRs (OzVRs) in the female muskrat and elucidate their physicochemical properties and evolutionary relationship. Six predominantly expressed OzVR genes were identified using the RACE technique, and a comprehensive analysis was conducted on their gene structure, subcellular distribution, functional predictions, and mRNA levels, revealed that all OzVRs were transmembrane proteins. Phylogenetic analysis clustered OzVR genes into two clades (V1Rs: OzV1R21, OzV1R81, OzV1R105; V2Rs: OzV2R33, OzV2R44, OzV2R60). Physiochemically, OzV1Rs were basic proteins, while OzV2Rs exhibited weakly acidic character. Among them, OzV1R81 and OzV2R44 were identified as hydrophobicitystable proteins, with the remainder categorized as hydrophobicity-unstable proteins. Promoters analysis revealed the involvement of transcription factors and complexes, including Ahr::Arnt, Runx1, Arnt, and TFAP2A, in regulating the expression of the OzVR genes. Conserved domain and motif analyses demonstrated a high conservation of the VRs superfamily in rodents, with many conserved domains linked to pheromone binding. Functional predictions confirmed that OzVRs were associated with pheromones detection. Finally, the expression patterns of OzVR genes in different tissues and seasons indicated that OzVRs have the highest level of expression in the vomeronasal organ, and OzV1Rs notably higher in the breeding season than that in the non-breeding season, however the expression levels of OzV2Rs were higher in the non-breeding season. This study provided insights into the phylogenetic relationships, gene structure, physicochemical properties, promoter binding sites, functions and gene expression patterns of OzVRs, offering a theoretical reference for further examination of VR gene functions and a foundation for understanding chemical signaling mechanisms in the muskrat.

Quantifying the role of iron recycling by Adélie and Emperor penguins over the austral spring and summer in Prydz Bay

In large areas of the Southern Ocean, iron limits phytoplankton production. Although biologically mediated iron recycling has been studied for the higher trophic-level whales and the lower trophic-level krill, less is known of the numerically abundant seabirds foraging in Antarctic waters. In this study, we estimate the magnitude of iron recycled by two Antarctic breeding seabirds, the Adélie and emperor penguins, across the austral spring and summer in the Prydz Bay region, East Antarctica. Their contribution to iron recycling and associated pathways differs in line with their contrasting life history strategies (summer and winter breeding) and their breeding habitat (land and fast ice). We consider their breeding cycle in relation to their terrestrial activities compared to foraging periods at sea. High iron concentration (~419 mg kg−1) in guano of both penguin species suggests that they are a source of regenerated iron. Breeding emperor penguins supplied an average of 237 μmol iron m−2 day−1 on the fast ice that they breed on that eventually ends in the ocean when the ice melts completely in summer (November–February). During their foraging trips, the adult emperor penguins contribute between 7 × 10−5 and 4 × 10−4 μmol iron m−2 day−1, as their foraging ranges increase over the breeding season. In contrast, breeding Adélie penguins supplied between 254 and 1,243 μmol iron m−2 day−1 whilst at their colony, with a fraction of guano entering the ocean via meltwater flowing into the ocean. The flux decreases to 2 × 10−3 to 6 × 10−2 μmol iron m−2 d−1, whilst they are foraging. Our study finds that penguins redistribute a large flux of iron onto their colonies, which may enter the adjacent water through sea ice melt and facilitated through katabatic winds. Despite their high abundance in Prydz Bay, the contribution of penguins to iron flux during their foraging periods is minor, due to the enormous foraging range being covered. Further research into the bioavailability of iron by marine organisms coupled with parallel measurements of seawater iron concentration and phytoplankton uptake experiments will be invaluable in refining iron budgets in both this region and other hotspots along the Antarctic coast where higher trophic-level animals are abundant.

Seasonal influence on miRNA expression dynamics of extracellular vesicles in equine follicular fluid.

Ovarian follicular fluid (FF) is a dynamic environment that changes with the seasons, affecting follicle development, ovulation, and oocyte quality. Cells in the follicles release tiny particles called extracellular vesicles (EVs) containing vital regulatory molecules, such as microRNAs (miRNAs). These miRNAs are pivotal in facilitating communication within the follicles through diverse signaling and information transfer forms. EV-coupled miRNA signaling is implicated to be associated with ovarian function, follicle and oocyte growth and response to various environmental insults. Herein, we investigated how seasonal variations directly influence the ovulatory and anovulatory states of ovarian follicles and how are they associated with follicular fluid EV-coupled miRNA dynamics in horses. Ultrasonographic monitoring and follicular fluid aspiration of preovulatory follicles in horses during the anovulatory (spring: non-breeding) and ovulatory (spring, summer, and fall: breeding) seasons and subsequent EV isolation and miRNA profiling identified significant variation in EV-miRNA cargo content. We identified 97 miRNAs with differential expression among the groups and specific clusters of miRNAs involved in the spring transition (miR-149, -200b, -206, -221, -328, and -615) and peak breeding period (including miR-143, -192, -451, -302b, -100, and let-7c). Bioinformatic analyses showed enrichments in various biological functions, e.g., transcription factor activity, transcription and transcription regulation, nucleic acid binding, sequence-specific DNA binding, p53 signaling, and post-translational modifications. Cluster analyses revealed distinct sets of significantly up- and down-regulated miRNAs associated with spring anovulatory (Cluster 1) and summer ovulation-the peak breeding season (Clusters 4 and 6). The findings from the current study shed light on the dynamics of FF-EV-coupled miRNAs in relation to equine ovulatory and anovulatory seasons, and their roles in understanding the mechanisms involved in seasonal shifts and ovulation during the breeding season warrant further investigation.

Influence of Sires on Population Substructure in Dülmen Wild Horses.

The objectives of the present study were to analyze the influence of the stallions employed in the Dülmen wild horses on the genetic diversity and population substructure using Bayesian cluster analysis. The Dülmen wild horse is maintained as a unique horse population exposed to the natural conditions all year round in the Merfelder Bruch near Dülmen in Westphalia, Germany. Stallions selected for breeding have to prove their abilities to survive under this harsh environment. We used multilocus genotypic information from a set of 29 autosomal microsatellites to determine the paternity of 185 male foals sired by nine stallions. As females could not be sampled, we could not make inferences on all yearlings and test whether there are differences in the genetic population parameters between both sexes. The mean number of progeny was 19.92 with a range of 2-32, caused by the length of the service period per stallion. The average observed and unbiased expected heterozygosity was 0.688 and 0.631, the mean number of alleles was 4.448, and Wright's FIS was -0.173. Pairwise genetic distances (FST and Nei's unbiased genetic distances) were significant and varied between 0.038 to 0.091 and 0.085 to 0.290, respectively. Neighbor-joining dendrogram plots clustered a large proportion of the paternal progeny groups in different branches. Posterior Bayesian analyses using seven paternal half-sib groups with 10-74 members supported a maximum of six clusters, with two paternal progeny groups not differing, and a median of five clusters, with two groups of two sires each falling into the same clusters. When sires were employed in non-consecutive years, progeny from these different years of the same sires were grouped in the same cluster, whereas the progeny of one sire from two consecutive years were in different clusters. We were able to distinguish male progeny from Dülmen wild horse stallions and to show the effects of stallion use on the genetic substructure in the Dülmen wild horse herd. In conclusion, the analyses showed the genetic potential of the Dülmen wild horse stallions to maintain a high genetic diversity and also the effects in which breeding seasons and for how long stallions are used to sire foals. The selection of stallions may be sensitive for the further development of genetic diversity and preserve this closed population as a valuable resource for further studies on the evolution of the horse.

Flammulated Owls Exhibit Diel Variation in Space Use

ABSTRACT A home range determines the resources animals can access, and the size of a home range often reflects resource availability and energetic requirements of individuals. The Flammulated Owl (Psiloscops flammeolus) is a cryptic small forest owl that breeds in mixed conifer forests in western North America. The home ranges of individuals generally comprise open forests with large trees, but we have yet to fully understand temporal variation in this species’ use of space and the habitat structures that drive space use at a fine scale. During the 2017 summer breeding season (May–July), we tracked the movement of six territorial males with GPS tags to estimate temporal variation in space use and examined resource selection for fine-scale habitat characteristics within home ranges. Results suggest that individual movement was more constrained around nests early at night, but the area of space used was consistent between the incubation and nestling stages. The owls’ activity centers near their nests had denser ground cover than available habitat, indicating that the forest understory may be valuable for this species’ breeding ecology. As forested landcover rapidly changes in western North America, understanding the spatial behavior and fine scale habitat associations of this species and others is increasingly important.