First record of the coypu Myocastor coypus (Molina, 1782)for the Arthur Thomas Municipal Park in themunicipality of Londrina, Paraná / Primeiro registro de ratão-do-banhado Myocastor coypus(Molina, 1782) para o Parque Municipal ArthurThomas no município de Londrina, Paraná

Myocastor coypus (coypu or nutria) is considered one of the 100 worst invasive species in the world due to its risk to local wildlife, such as waterfowl, zoonotic risks, and environmental damage, such as riverbank erosion, arising from its habit of constructing burrows along the edge of water bodies. The presence of M. coypus is already known locally in the municipality of Londrina based on records at Igapó Lake. This paper presents the first record of M. coypus in Arthur Thomas Municipal Park, a Conservation Unit of Integral Protection located in the urban area of Londrina. The records were obtained through direct observation of one live individual and one carcass during campaigns to monitor medium and large mammals. Subsequently, ten records were obtained using camera traps, of unknown gender, located near waterbodies. We emphasize the need for continuous fauna monitoring in conservation units to detect and verify potential increases in invasive alien species populations that can result in environmental damage.

A espécie Myocastor coypus (ratão-do-banhado) é considerada uma das 100 piores espécies invasoras do mundo por representar risco para a fauna local, como aves aquáticas, e risco de zoonoses, além de causar danos ambientais, como desbarrancamento de encostas de rios, devido ao seu hábito de construção de tocas nas margens de corpos d'água. A presença de M. coypus já é conhecida localmente no município de Londrina a partir de registros no Lago Igapó. Assim, este estudo apresenta o primeiro registro de M. coypus no Parque Municipal Arthur Thomas, uma Unidade de Conservação de Proteção Integral localizada na zona urbana de Londrina. Os registros foram obtidos através da observação direta de um indivíduo vivo e uma carcaça durante campanhas de monitoramento de mamíferos de médio e grande porte. Posteriormente foram obtidos dez registros por armadilha fotográfica de indivíduos de sexo desconhecido, próximos a corpos d'água. Enfatiza-se a necessidade do monitoramento contínuo da fauna em áreas de conservação para a detecção do potencial aumento na população de espécies exóticas e invasoras que podem causar severos danos ambientais.

Congruence between co-occurrence and trait-based networks is scale-dependent: a case study with flea parasites of small mammalian hosts.

We applied a novel framework based on network theory and a concept of modularity that estimates congruence between trait-based ( = functional) co-occurrence networks, thus allowing the inference of co-occurrence patterns and the determination of the predominant mechanism of community assembly. The aim was to investigate the relationships between species co-occurrence and trait similarity in flea communities at various scales (compound communities: across regions within a biogeographic realm or across sampling sites within a geographic region; component communities: across sampling sites within a geographic region; and infracommunities: within a sampling site). We found that compound communities within biogeographic realms were assembled via environmental or host-associated filtering. In contrast, functional and spatial/host-associated co-occurrence networks, at the scale of regional compound communities, mostly indicated either stochastic processes or the lack of dominance of any deterministic process. Analyses of congruence between functional and either spatial (for component communities) or host-associated (for infracommunities) co-occurrence networks demonstrated that assembly rules in these communities varied among host species. In component communities, stochastic processes prevailed, whereas environmental filtering was indicated in 4 and limiting similarity/competition in 9 of 31 communities. Limiting similarity/competition processes dominated in infracommunities, followed by stochastic mechanisms. We conclude that assembly processes in parasite communities are scale-dependent, with different mechanisms acting at different scales.

[Regulation of Transcription by RNA Polymerase III Promotors in the Norm and Pathology].

RNA polymerase III synthesizes a wide range of noncoding RNAs shorter than 400 nucleotides in length. These RNAs are involved in protein synthesis (tRNA, 5S rRNA, and 7SL RNA), maturation, and splicing of different types of RNA (RPR, MRP RNA, and U6 snRNA), regulation of transcription (7SK RNA), replication (Y RNA), and intracellular transport (vault RNA). BC200 and BC1 RNA genes are transcribed by RNA polymerase III in neurons only where these RNAs regulate protein synthesis. Mutations in the regulatory elements of the genes transcribed by RNA polymerase III as well as in transcription factors of this RNA polymerase are associated with the development of a number of diseases, primarily oncological and neurological. In this regard, the mechanisms of regulation of the expression of the genes containing various RNA polymerase III promoters were actively studied. This review describes the structural and functional classification of polymerase III promoters, as well as the factors involved in the regulation of promoters of different types. A number of examples demonstrate the role of the described factors in the pathogenesis of human diseases.

Viral metagenomic survey of Caspian seals.

Introduction: Viral diseases of pinnipeds cause substantial mortality and morbidity and can influence population demography. Viral metagenomic studies can therefore play an important role in pinniped health assessments and disease surveillance relevant to both individual species and in a "One Health" context. Methods: This study used a metagenomic approach with high throughput sequencing to make the first assessment of viral diversity in Caspian seals (Pusa caspica), the only marine mammal species endemic to the Caspian Sea. Results: Sequencing libraries from 35 seals sampled 2009-2020 were analysed, finding sequences from the viral families Circoviridae, Parvoviridae, Herpesviridae, Papillomaviridae, Picornaviridae, Caliciviridae, Cruciviridae, Anelloviridae, Smacoviridae, and Orthomyxoviridae, with additional detection of Adenoviridae via PCR. The similarity of viral contigs from Caspian seal to sequences recovered from other pinnipeds ranged from 63.74% (San Miguel sea lion calicivirus) to 78.79% (Seal anellovirus 4). Discussion: Some findings represent novel viral species, but overall, the viral repertoire of Caspian seals is similar to available viromes from other pinnipeds. Among the sequences recovered were partial contigs for influenza B, representing only the second such molecular identification in marine mammals. This work provides a foundation for further studies of viral communities in Caspian seals, the diversity of viromes in pinnipeds more generally, and contributes data relevant for disease risk assessments in marine mammals.

Combination of in silico and molecular techniques for discrimination and virulence characterization of marine Brucella ceti and Brucella pinnipedialis.

Introduction: Mammals are the main hosts for Brucella sp., agents of worldwide zoonosis. Marine cetaceans and pinnipeds can be infected by Brucella ceti and B. pinnipedialis, respectively. Besides classical bacteriological typing, molecular approaches such as MLVA, MLSA, and whole-genome sequencing (WGS) can differentiate these species but are cumbersome to perform. Methods: We compared the DNA and genome sequences of 12 strains isolated from nine marine mammals, with highly zoonotic B. melitensis, B. abortus, and B. suis, and the publicly available genomes of B. ceti and B. pinnipedialis. In silico pipelines were used to detect the antimicrobial resistance (AMR), plasmid, and virulence genes (VGs) by screening six open-source and one home-made library. Results and discussion: Our results show that easier-to-use HRM-PCR, Bruce-ladder, and Suis-ladder can separate marine Brucella sp., and the results are fully concordant with other molecular methods, such as WGS. However, the restriction fragment length polymorphism (RFLP) method cannot discriminate between B. pinnipedialis and B. ceti B1-94-like isolates. MLVA-16 results divided the investigated strains into three clades according to their preferred host, which was confirmed in WGS. In silico analysis did not find any AMR and plasmid genes, suggesting antimicrobial susceptibility of marine Brucella, while the presence of the VGs btpA gene was variable dependent on the clade. Conclusion: The HRM-PCR and Suis-ladder are quick, easy, and cost-effective methods to identify marine Brucella sp. Moreover, in silico genome analyses can give useful insights into the genetic virulence and pathogenicity potential of marine Brucella strains.

The signal that stimulates mammalian embryo development.

Embryo development is stimulated by calcium (Ca2+) signals that are generated in the egg cytoplasm by the fertilizing sperm. Eggs are formed via oogenesis. They go through a cell division known as meiosis, during which their diploid chromosome number is halved and new genetic combinations are created by crossing over. During formation the eggs also acquire cellular components that are necessary to produce the Ca2+ signal and also, to support development of the newly formed embryo. Ionized calcium is a universal second messenger used by cells in a plethora of biological processes and the eggs develop a "toolkit", a set of molecules needed for signaling. Meiosis stops twice and these arrests are controlled by a complex interaction of regulatory proteins. The first meiotic arrest lasts until after puberty, when a luteinizing hormone surge stimulates meiotic resumption. The cell cycle proceeds to stop again in the middle of the second meiotic division, right before ovulation. The union of the female and male gametes takes place in the oviduct. Following gamete fusion, the sperm triggers the release of Ca2+ from the egg's intracellular stores which in mammals is followed by repetitive Ca2+ spikes known as Ca2+ oscillations in the cytosol that last for several hours. Downstream sensor proteins help decoding the signal and stimulate other molecules whose actions are required for proper development including those that help to prevent the fusion of additional sperm cells to the egg and those that assist in the release from the second meiotic arrest, completion of meiosis and entering the first mitotic cell division. Here I review the major steps of egg formation, discuss the signaling toolkit that is essential to generate the Ca2+ signal and describe the steps of the signal transduction mechanism that activates the egg's developmental program and turns it into an embryo.

Alterations of pleiotropic neuropeptide-receptor gene couples in Cetacea.

BACKGROUND: Habitat transitions have considerable consequences in organism homeostasis, as they require the adjustment of several concurrent physiological compartments to maintain stability and adapt to a changing environment. Within the range of molecules with a crucial role in the regulation of different physiological processes, neuropeptides are key agents. Here, we examined the coding status of several neuropeptides and their receptors with pleiotropic activity in Cetacea. RESULTS: Analysis of 202 mammalian genomes, including 41 species of Cetacea, exposed an intricate mutational landscape compatible with gene sequence modification and loss. Specifically for Cetacea, in the 12 genes analysed we have determined patterns of loss ranging from species-specific disruptive mutations (e.g. neuropeptide FF-amide peptide precursor; NPFF) to complete erosion of the gene across the cetacean stem lineage (e.g. somatostatin receptor 4; SSTR4). CONCLUSIONS: Impairment of some of these neuromodulators may have contributed to the unique energetic metabolism, circadian rhythmicity and diving response displayed by this group of iconic mammals.

Correlation between mammal track abundance and Forest Landscape Integrity Index validates actual forest ecological integrity.

Human disturbance compromises the ecological integrity of forests, negatively affecting associated species. Assessing the impact of forest integrity on biodiversity is complex due to the interplay of various human activities, ecological factors, and their interactions. Current large-scale indices assess forest integrity but often lack a direct connection to the biotic environment. We tested the effectiveness of the global Forest Landscape Integrity Index (FLII) in evaluating aspects of anthropogenic forest degradation on the biotic community. We analyzed the relationship between changes in the ecological integrity of Finnish forests and variations in mammal species abundance, using the number of tracks from 17 different species collected during the winter seasons between 2016 and 2020 in south-central Finland. Beyond the FLII, we analyzed forest and canopy cover to enhance the accuracy of habitat preference assessments. We found that the FLII captures the varying degrees of forest integrity, as reflected by the correlation between the abundance of winter tracks and the FLII for most mammals. Species that were positively associated with forest integrity were all native to the boreal forest, while mammals that adapt well to human-disturbed environments including two invasive species were more common in lower FLII forests. Significant differences in habitat preferences were also observed in relation to forest and canopy cover, revealing additional nuances that the FLII alone did not capture. This study demonstrates that the FLII, when combined with a comprehensive dataset and supplemented with region-specific factors, can assess species' adaptability to human-modified forests, aiding in the development of conservation strategies.

Anatomical description of a pygmy sperm whale, Kogia breviceps (Cetacea: Kogiidae), pre-term calf using CT scan and 3D reconstructions.

Little is known about the biology of pygmy sperm whales, Kogia breviceps (De Blainville, 1838), being that most anatomical descriptions for the species derive from necropsy after stranding or from osteological material preserved in museums. This species is rarely seen despite its wide distribution, and its reproductive behaviour is still being investigated. The eventual occurrence of pregnant female strandings and the collection and description of foetuses can give clues about the organisms' mostly unknown early development. However, this type of biological material is extremely rare, limiting anatomical analysis due to the risk of damage or loss. Here, we describe the external and internal anatomy of an 84 cm long K. breviceps foetus. The methods utilised were non-intrusive, meaning that no incisions were made on the specimen. The foetus was analysed using computed tomography images and a three-dimensional reconstruction of the skeleton. A great number of features were observed, such as axial and appendicular skeletal structures, internal organs, echolocation apparatus and umbilical cord, as well as diagnostic characters of the species, such as the asymmetrical skull, spermaceti chamber and false gill pigmentation. We suggest that more specimens on different stages of development should be analysed by the same technique, as well as further comparison with specimens from other taxa, in order to facilitate more comparative studies on embryonic and foetal development of cetaceans.

Molecular-phylogenetic analyses of Babesia and Theileria species from small mammals and their ticks in northern China suggest new reservoirs of bovine and equine piroplasms.

Babesia and Theileria species (Apicomplexa: Piroplasmida) are tick-borne protozoan parasites that can cause mild to severe infection in humans, wildlife, livestock and companion animals. To date, reports on the molecular study of piroplasms from wild living small mammals and their ticks are still limited, especially in Asia. This study encompassed an extensive survey involving 907 liver samples and 145 ixodid ticks from 16 different species of small mammals (Rodentia, Lagomorpha, Eulipotyphla). These were collected in 13 cities and counties in northern China. DNA extracts from these samples were screened for the presence of piroplasm 18S rRNA gene. Samples that tested positive were further evaluated for other genetic markers of piroplasms, including the cox1 gene and the ITS1-5.8S rDNA-ITS2 region. Several piroplasm species were identified, including Babesia sp. tavsan2, Babesia occultans, Theileria sp. Xinjiang, Theileria equi, and Theileria sp. Kalecik. Among these, Theileria sp. Xinjiang was shown to be the most prevalent. Importantly, Babesia sp. tavsan2 was identified in the tick Rhipicephalus sanguineus from the Yarkand hare and Theileria sp. Kalecik in Hyalomma asiaticum from the long-eared hedgehog, in line with the detection of these pathogens in tissue samples of the relevant hosts. This study further disclosed the presence of DNA from B. occultans and T. equi, typically found in cattle and horses respectively, with an additional discovery in small mammals. Moreover, Theileria sp. Kalecik, which was first detected in small-sized mammals, and Babesia sp. tavsan2, were both reported for the first time in China.

Potential biosecurity breaches in poultry farms: Presence of free-ranging mammals near laying-hen houses assessed through a camera-trap study.

Diligent application and implementation of biosecurity measures stand as the most effective measures to prevent disease transmission through direct or indirect interactions between poultry and free-ranging animals. Among these, free-ranging mammals can be hosts or disseminators of several pathogens relevant to poultry and of public health concern. Moreover, evidence of susceptibility to avian influenza virus infection in non-human mammals has raised questions about their potential role in the virus' epidemiology at the domestic animal-wildlife interface. Given this background, this study aimed to identify mammal species occurring near laying-hen houses and characterize the spatiotemporal patterns of these visits. Seven camera traps were deployed for a year-long period in three commercial poultry farms in a densely populated poultry area in Northern Italy. Various methods, including time series analysis and generalized linear models, were employed to analyze daily mammal visits. A total of 1,867 camera trap nights yielded 567 videos of seven species of wild mammals, and 1,866 videos showed domestic pet species (cats and dogs). Coypus (Myocastor coypus) and cats were the two mammals more frequently observed near poultry houses. For wild mammals, visits significantly increased at night, and slightly decreased during the spring season. Overall, the data hereby provided lay the groundwork for designing novel surveillance and intervention strategies to prevent cross-species disease transmission. Moreover, the utilization of visual evidence depicting free-ranging animals approaching poultry houses could assist health authorities in educating and raising awareness among stakeholders about potential risks of pathogen spillover.

Increasing spillover of highly pathogenic avian influenza A (H5N1) virus to mammals.

•1.Increasing spillover of highly pathogenic avian influenza A (H5N1) virus to sea mammals.•2.Increasing spillover of highly pathogenic avian influenza A (H5N1) virus to fur mammals.•3.Increasing spillover of highly pathogenic avian influenza A (H5N1) virus to ruminant animals.•4.Cross-species transmission of highly pathogenic avian influenza A (H5N1) virus between ruminant animals and humans.•5.Highly pathogenic avian influenza A (H5N1) virus RNA was present in milk.

Comparative study of the oligosaccharide profile in goat, bovine, sheep, and human milk whey.

Milk oligosaccharides are high added value compounds that could be obtained by exploiting cheese whey, a byproduct of dairy industry. The objective was to compare the abundance and diversity of oligosaccharides in whey samples from domestic animals and humans. During fresh cheese making, whey samples were collected and analyzed by untargeted and targeted small molecule analysis using high-resolution mass-spectrometry. A great similarity in the metabolite profile between goat and sheep was observed. Up to 11 oligosaccharides were observed in the sheep whey from those typically found in humans. The concentration of 2'-Fucosyllactose (0.136 ± 0.055 g/L) and 3-Fucosyllactose (0.079 ± 0.009 g/L) were significantly higher (p-value <0.01) in sheep whey, while the concentration of 3'-Sialyllactose (0.826 ± 0.638 g/L) was higher in goat whey. No significant differences were observed between goat and sheep whey for the other oligosaccharides (p-value >0.05). Therefore, sheep and goat whey could become an important source of oligosaccharides through their revalorization.

Progress on SOX9 and its enhancers in mammalian sex determination.

The sex determination in mammals refers to the development of an initial bipotential organ, termed the bipotential gonad/genital ridge, into either a testis or an ovary at the early stages of embryonic development, under the precise regulation of transcription factors. SOX9 (SRY-box transcription factor 9) is a multifunctional transcription factor in mammalian development and plays a critical role in sex determination and subsequent male reproductive organs development. Recent studies have shown that several enhancers upstream of SOX9 also play an important role in the process of sex determination. In this review, we summarize the progress on the role of SOX9 and its gonadal enhancers in sex determination. This review will facilitate to understand the regulatory mechanism of sex determination of SOX9 and provides a theoretical basis for the further development of animal sex manipulation technologies.

Invariance of Mitochondria and Synapses in the Primary Visual Cortex of Mammals Provides Insight Into Energetics and Function.

The cerebral cortex accounts for substantial energy expenditure, primarily driven by the metabolic demands of synaptic signaling. Mitochondria, the organelles responsible for generating cellular energy, play a crucial role in this process. We investigated ultrastructural characteristics of the primary visual cortex in 18 phylogenetically diverse mammals, spanning a broad range of brain sizes from mouse to elephant. Our findings reveal remarkable uniformity in synapse density, postsynaptic density (PSD) length, and mitochondria density, indicating functional and metabolic constraints that maintain these fundamental features. Notably, we observed an average of 1.9 mitochondria per synapse across mammalian species. When considered together with the trend of decreasing neuron density with larger brain size, we find that brain enlargement in mammals is characterized by increasing proportions of synapses and mitochondria per cortical neuron. These results shed light on the adaptive mechanisms and metabolic dynamics that govern cortical ultrastructure across mammals.

Biomagnification potential and health risks of organophosphate esters in prey to humpback dolphins based on dietary correction.

Information on the biomagnification of organophosphate esters (OPEs) is limited, and the results are inconclusive, mainly because precise predatorprey relationships have not been determined. Herein, we first evaluated the biomagnification potential and dietary exposure risk of 15 OPEs in 14 prey species (n = 234) to Indo-Pacific humpback dolphins from the northern South China Sea using quantitative fatty acid signature analysis (QFASA). QFASA identified Chinese gizzard shad as the primary prey of dolphins. Among the 15 OPEs, 86.7 % (13/15) had a diet-adjusted biomagnification factor (BMFQFASA) greater than 1, indicating the biomagnification potential between dolphins and their diet. Moreover, BMFQFASA exhibited a considerable positive correlation with the log octanolwater partitioning coefficient of OPEs, indicating that lipophilicity may affect the bioamplification of OPEs. Risk assessments showed that although current OPE levels may not pose substantial health risks to dolphins via diet intake, the nondiet-adjusted hazard quotient/hazard index underestimated the exposure risk of OPEs to this vulnerable dolphin species. This study provides novel evidence regarding the biomagnification and dietary exposure risks of OPEs in cetaceans, emphasizing the importance of estimating the dietary composition of predators in such analyses.

New Additions to the Mammal List Documented in the Portuguese Red Data Book.

This study outlines the procedures used for collecting, processing, and categorizing data on 16 new mammal species for mainland Portugal, belonging to four taxonomic groups: Eulipotyphla (1), Chiroptera (4), Rodentia (2), and Cetacea (9). Data collection and processing encompassed field and lab work and bibliographic compilation. Data categorization involves, whenever possible, the assessment of the approximate number of mature individuals in populations, the extent of occurrence, and the area of occupancy. The approach employed led to the classification of eight out of the 16 species into an IUCN category: two non-volant small mammals and one bat species were designated as Vulnerable, requiring ongoing monitoring; one rodent and three cetaceans were assigned to Data Deficient due to insufficient available information; and a single bat species was classified as Least Concern due to the high abundance of local populations. For small mammals and bats, alterations to natural systems and climate change emerged as the most relevant threatening factors, while for cetaceans, human activities such as fishing, commercial shipping, and tourism were identified as the primary survival risks. It is recommended to maintain action programs that assist in defining strategic orientations for the implementation of conservation measures on a case-by-case basis.

Dietary Shift in a Barn Owl (Tyto alba) Population Following Partial Abandonment of Cultivated Fields (Central Apennine Hills, Italy).

While most studies focused on the impact of intensive agriculture on the barn owl's diet, little is known about the effect of cropland abandonment. We compared the taxon composition/evenness and feeding guild structure of small mammal prey identified in pellets collected before (2004) and after (2012) the abandonment of 9% of cultivated fields within a cultural landscape. Data on prey abundance per pellet were analysed through non-metric multidimensional scaling and permutational, paired tests. Prey taxon evenness in 2012 was significantly lower than in 2004. That induced a shift in prey taxon composition as indicated by the significantly lower dietary similarity compared with the random expectation. The increasing and declining abundance of Murinae and Crocidurinae, respectively, had the largest contribution to the differentiation of the diet spectrum. Insectivorous prey was significantly more abundant in 2004 compared to 2012, while the opposite was true for omnivorous prey. Our results suggest that even a small fraction of abandoned crops in the landscape might induce a detectable shift in the barn owl's food niche. The dietary effects are similar to those observed after agricultural intensification, that is, an increase in the abundance of generalists to the detriment of specialist mammal prey.

Human activities reshape the spatial overlap between North Chinese leopard and its wild ungulate prey.

BACKGROUND: Rapidly expanding human activities have profoundly changed the habitat use of both large carnivores and their prey, but whether and how human activities affect the interactions between them has received relatively less attention. In this study, we conducted a systematically designed camera-trapping survey on an endangered large carnivore (North Chinese leopard Panthera pardus japonensis) and its wild ungulate prey (Siberian roe deer Capreolus pygargus and wild boar Sus scrofa) in the Taihang Mountains of central North China. Using conditional two-species occupancy model based on data derived from the extensive sampling effort (15,654 camera-days at 102 camera sites), we examined the relationship of spatial use between leopards and each prey species under the effects of human presence, free-ranging cattle, roads and settlements. RESULTS: Humans and cattle had contrasting effects on the relationship of spatial use between leopard and roe deer, with higher and lower spatial segregation between them at human and cattle-frequented sites, respectively. Roads might create a shelter for wild boar from leopard predation, with less spatial segregation between them at sites close to the roads. CONCLUSIONS: Our findings demonstrate that human activities are reshaping the spatial overlap between large carnivores and their prey, and have non-equivalent effects among different types of human activity. Such effects may further alter the strength of interspecific interactions between predator and prey, with far-reaching influences on the community and ecosystem that require more research.

An assessment of seasonal bait uptake by individual grey squirrels to develop a delivery system for oral contraceptives.

Globally, human-wildlife conflicts continue to increase, owing to human population growth and expansion. Many of these conflicts concern the impacts of invasive non-native species. In the UK, the invasive, non-native grey squirrel Sciurus carolinensis negatively affects tree health and has caused the decline of the native red squirrel Sciurus vulgaris. Oral contraceptives are being developed to manage the impacts of the grey squirrel. To be effective, contraceptives will need to be deployed at a landscape scale, and will require a delivery system that is practical and economically viable. Understanding grey squirrel feeding behaviour is important so that delivery methods can be designed so that a sufficient number of target individuals receive an effective contraceptive dose at a time of year that will ensure their infertility throughout peak times of breeding. The main aims of this study were to assess how sex, season, squirrel density and bait point density influenced; (1) the probability of a squirrel visiting a feeder and (2) the amount of bait consumed from feeders. Field trials were conducted on six woodland populations of squirrels in three seasons, with four days of bait deployment via purpose-designed squirrel-specific bait hoppers with integrated PIT-tag readers. It was possible to deliver multiple doses on most days to most male and female grey squirrels, with bait deployment more likely to be effective in spring, immediately before the second annual peak in squirrel breeding, followed by winter, immediately before the first peak in breeding. The results from this study could be used to design methods for delivering oral contraceptive baits to grey squirrels in the future and the methods used could be applied to other small mammal species and other bait delivery systems. © 2024 Crown copyright and The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. This article is published with the permission of the Controller of HMSO and the King's Printer for Scotland.