Lab Animal

Cooperation in social species is shaped by ongoing social relationships, partner choice, and group interactions, demanding experimental systems that preserve the social context in which these behaviors unfold. Here we introduce the e-Seesaw, a wireless system for automated liquid reward delivery designed to support home-enclosure experiments on reward access distribution in common marmosets (Callithrix jacchus) with minimal human intervention. The apparatus combines a modular peristaltic pump with a Bluetooth-controlled trigger, allowing spatial separation between the site of action and the site of reward delivery while preserving group housing. We provide detailed design files, software, and assembly instructions to support reproduction and adaptation. In a proof-of-concept deployment across seven families, animals readily engaged with the device, producing a median of ~87 trigger activations per session. Engagement was concentrated early within sessions and remained largely stable across repeated deployments, including under increased action-reward separation. These results established the e-Seesaw as a flexible and reproducible platform for automated reward-delivery experiments in animals tested within their social groups, while reducing human involvement and avoiding fixed dyadic testing.

Background: Canadian biomedical core facilities (BCFs) provide researchers with access to advanced tools and unique technical expertise, essential for research. However, their role, sustainability, and impact remain poorly understood. We report on the evolution of model and existing state of Canadas 205 BCFs, examining challenges and benefits. Methods: Cross referencing of national databases by the Canadian Innovation Fund (CFI), and from data collected by the Canadian Network of Scientific Platforms (CNSP) allowed he identification of BCFs. Hand curation of these lists validated that cores are operational. To ensure cores not listed by CFI/CNSP were captured, research intensive institutions in Canada were independently searched to identify active cores. Results: There are currently 205 active and operational BCFs located across 9 provinces, which can be further stratified into 9 technical domains that describe the nature of services they provide. Quebec (80 cores) and Ontario (75 cores) have the highest confluence of BCFs, with Quebec having a higher ratio of cores per capita. Conclusions: While our data establishes the ubiquity of Canadian BCFs, we highlight substantial challenges including sustainability, governance, evaluation and the recognition of support for core scientists. Here, we establish a framework to address these challenges and to inform best practice, to optimize creation of impactful, accessible and functional biomedical core facilities.

Background/ObjectivesThe integration of large language model (LLM)-based AI tools into veterinary clinical practice is rapidly increasing; however, no systematically derived taxonomy of veterinary AI query patterns has been established. This study aimed to develop and refine an exploratory taxonomy of veterinary AI query patterns across clinical stages through a structured expert-panel review process. MethodsAn exploratory cross-sectional expert panel study was conducted. 5,372 real-world query logs from a veterinary clinical AI chatbot deployed over eight months were analyzed using AI-assisted inductive coding to derive an initial taxonomy. The taxonomy was refined through literature review and subsequently reviewed by an expert panel of 38 veterinary professionals via structured online survey across five clinical stages. ResultsA taxonomy of 3 categories and 21 subtypes was established: Clinical Support Queries (Types A-H), Evidence-Based Research Queries (Types I-L), and Terminology and Drug Reference Queries (Types M-U). Type B (Differential Reasoning) had the highest overall frequency (57/188 first-choice responses), while Type D (Clinical Decision Support) was dominant immediately post-consultation (55.3%). Veterinary professionals with [&ge;]10 years of experience showed a higher frequency of Type G (Evidence Search) preference than those with <10 years of experience (18 vs. 4), while university-affiliated professionals demonstrated a distinct pattern dominated by Type G. ConclusionsTo our knowledge, no published study has previously established a veterinary-specific, clinical-stage-sensitive exploratory taxonomy of AI query patterns; this study addresses that gap. The findings provide a foundational framework for designing context-aware, stage-adaptive veterinary AI systems and benchmark evaluation tools.

Retinal and choroidal vascular diseases are major causes of vision loss that require frequent intravitreal anti-VEGF therapy. Anti-angiogenic peptide AXT107 demonstrated efficacy in preclinical studies and was advanced to the clinical stage. To provide for sustained delivery of the peptide and avoid complications with intravitreal injection, we evaluated suprachoroidal delivery of AXT107 microparticles (MP-AXT107). The original, soluble AXT107 formulation was ineffective at inhibiting laser-induced choroidal neovascularization (CNV) in our rat model and was consequently reformulated as microparticles. MP-AXT107 demonstrated high peptide incorporation efficiency, reproducible morphology, and physical and chemical stability for at least 9 months under refrigerated storage. In the rat CNV model, suprachoroidal MP-AXT107 significantly reduced neovascular area by approximately 60% relative to vehicle controls. Safety and durability were evaluated in a 9-month GLP toxicology study in Gottingen minipigs following a single suprachoroidal injection of vehicle or MP-AXT107 (0.125-1.25 mg/eye). Transient increases in IOP and mild ocular inflammatory findings were observed immediately following administration but resolved rapidly without lasting effects. No treatment-related adverse ocular findings were observed during the remainder of the study, and the highest tested dose (1.25 mg/eye) was established as the no-observed-adverse-effect level. Bioanalysis at study completion demonstrated persistent AXT107 localization primarily within choroid/RPE and scleral tissues, with no signs of systemic exposure. Collectively, these findings demonstrate that suprachoroidal delivery of MP-AXT107 enables sustained anti-angiogenic activity with favorable ocular safety and prolonged tissue retention, supporting further clinical development as a durable therapy for retinal and choroidal vascular diseases.

Accurate localization of anatomical landmarks is a foundational skill in anatomy and imaging that is often taught informally through expert mentorship, requiring access to data and desktop software. There is no openly accessible, interactive resource that teaches neuroanatomy with quantitative feedback. We present the AFIDs-Validator (validator.afids.io), an open-access, browser-based platform that pairs guided instruction with quantitative assessment. The platform combines (1) a learning mode in which a language-model neuroanatomy tutor operates inside an MRI viewer, giving anatomy-first instruction that responds to the learner's current image slice, orientation, and cursor position; and (2) a validation engine that accepts a learner's landmark file and returns per-landmark Euclidean error against expert-annotated references spanning 21 brain templates. To make the feedback interpretable, we analyzed 15,000 landmark annotations across 132 human subjects and found that landmark difficulty varies fourfold (median error ranged from 0.37 mm at the anterior commissure to 1.50 mm at the temporal horns) with heavy-tailed distributions at every landmark. These distributions are compiled into per-landmark reliability priors, so learners are scored against the empirical spread of trained raters rather than an arbitrary threshold, and difficult landmarks are not mistaken for poor performance. The AFIDs-Validator requires no installation, licensed software, or local data, and all code, reference data, and tutor design are openly released.

PurposeAccurate molecular characterisation of infection and inflammation within the distal human lung remains challenging, particularly in critically ill patients, due to limited access to the alveolar space and delayed diagnostic workflows. Molecular imaging approaches capable of real-time detection and targeted sampling could substantially improve diagnostic precision and the future translational development of molecular imaging probes and therapeutics. MethodsIn a preclinical setting, we evaluated a clinic-ready endomicroscopic fluorescence lifetime imaging microscopy (eFLIM) platform combined with molecularly targeted SmartProbes for in situ detection of bacteria and activated neutrophils in the distal human lung. A multifunctional 1.9-mm diameter imaging and sampling catheter (Eyes on Target; EoT) enabled real-time fluorescence intensity and lifetime imaging alongside directed alveolar microlavage via a 1.2-mm working channel. Fluorescence intensity and lifetime signatures of Gram-negative bacteria, Gram-positive bacteria, and activated neutrophils were characterised using three wash-free SmartProbes: NBD-PMX, Merocy-Van, and a neutrophil activation probe (NAP). Imaging and sampling performance were assessed in ventilated ex vivo human lungs. ResultsEoT reliably navigated to alveolar regions across all lung lobes in both phantom and ventilated human lung models. eFLIM distinguished alveolar microanatomy and enabled probe-specific molecular detection within the distal lung. Increased NBD-PMX signal was detected in Escherichia coli-instilled lobes, while Merocy-Van lifetime signatures selectively identified Staphylococcus aureus-instilled regions. Activated neutrophils were detected throughout lung tissue following NAP administration. Directed alveolar microlavage enabled recovery of cellular material and bacterial DNA from imaged regions for downstream analysis. ConclusioneFLIM using EoT combined with molecular SmartProbes enables real-time molecular imaging and targeted sampling within the distal human lung. This platform provides a translatable approach for evaluating infection and inflammation at the alveolar level and supports the clinical development of molecular imaging probes for pulmonary disease.

Operant paradigms are powerful tools to quantify motivation and reward. Traditionally, operant conditioning research has been limited to food and drug reinforcers. Recent advances in commercially available operant equipment, however, allow for the quantification of social motivation. These operant assays are an improvement over commonly used social preference tasks, as they enable direct measurement of the effort and motivation driving social behavior. Based on a design by Venniro et al. (2020), the MedPC social operant boxes modify the traditional operant box setup for social interactions. The experimental rat can lever-press to raise a door for an interaction with a target rat behind a porous barrier. These social operant boxes have been widely adapted to test social behavior in adult and adolescent rodents and investigate how a range of conditions (e.g. stress, drug taking, etc.) affect social motivation. However, there is a gap in assessing maternal motivation for pups during the postpartum period, despite ample evidence that postpartum social behavior is highly relevant for offspring health outcomes. Here, we detail 3D-printed modifications to the standard Med PC social operant boxes to adapt the social target chamber to safely house neonatal pups. We have also developed testing protocols to assess motivation during the limited postpartum period. These data demonstrate that, with simple modifications to social operant chambers and testing protocols, the field can implement advanced behavioral approaches to directly assess maternal motivation.

To facilitate the translation of NF1 exon 17 skipping as a mutation-specific therapy for Neurofibromatosis type 1 into in vivo testing, we have continued to develop more efficient antisense oligonucleotides (ASOs), humanized mouse models, and explored multiple delivery platforms including an adeno-associated virus (AAV)-U7-SnRNA vector approach. We evaluated both biodistribution and exon skipping efficacy of a U7-SnRNA targeting NF1 exon 17 with an SFFV-driven cassette containing T2A-linked Luciferase (Luc) and eGFP packaged in AAV-9, AAV-F and AAV-B1 capsids. We show that AAV-F is superior to AAV-9 and AAV-B1 for mouse brain delivery based on DNA transduction, GFP expression, and luciferase activity, but AAV-B1 delivers 2-4 fold more to sciatic nerve (SCN). In terms of exon skipping, AAV-F appears to induce the most skipping in liver and optic nerve (ON), while AAV-B1 mediates highest skipping in the liver, SCN, and ON. The identification of AAV serotypes that allow efficient transduction and delivery of transgenes to the mouse CNS and PNS is impactful for preclinical research in murine models of other diseases. Furthermore, this is both the first report of NF1 exon skipping efficacy in vivo and the first successful application of an U7-SnRNA for the restoration of functional neurofibromin for NF1.

Space biology literature is growing exponentially. Existing infrastructure has not kept pace with organizing, synthesizing, and disseminating this knowledge. We present SpaceBio SpaceBio Knowledge Hub (www.spacebio.space), an integrated digital ecosystem that combines artificial intelligence, real-time data integration, and open-access infrastructure to advance research, education, and collaboration in microgravity, space biology and space exploration. The platform applies AI-driven approaches including natural language processing, machine learning, and automated content generation to construct a semantic atlas of the field. The atlas reveals the hierarchical thematic organization underlying microgravity-induced biological responses, space mission infrastructure, planetary science, and astrobiology. As part of this effort, SpaceBio is moving toward the construction of a LiteratOmics framework for microgravity, and space biology a systematic, AI-enabled approach to mining, integrating, and structuring the primary literature generated by omics-driven spaceflight research, treating the scientific literature itself as a navigable data layer alongside genomic, transcriptomic, and proteomic datasets. Built on a scalable, cloud-based architecture with a user-centered interface, SpaceBio supports literature exploration, data integration, and knowledge discovery for researchers, educators, students, industry partners, and citizen scientists. The platform also functions as a community-building ecosystem. It integrates hands-on research initiatives, AI-generated educational content, pilot data science projects, and social responsibility programs that broaden participation without compromising scientific rigor. AI-enabled digital environments can transform fragmented literature into a navigable knowledge landscape. SpaceBio accelerates research productivity, strengthens STEM education, and supports the global space life sciences community as human space exploration enters in the most ambitious era.

Development of candidate countermeasures against human pathogens frequently includes nonhuman animal experimentation. Preclinical animal pathogen exposure studies are conducted to model diseases and accumulate preliminary and hypothetically translatable data to inform and justify the design of clinical trial evaluation of countermeasure safety and efficacy. In addition to frequent ethical critiques, challenges associated with animal experimentation include considerable resources needed to achieve statistical power and robustness, replicability and reproducibility concerns, potentially compromised objectivity through lack of blinding, fundamental species-specific biological differences, and risk of unpredictable pathogen adaptation to the experimental animal. Recent U.S. and U.K. government initiatives aim to reduce animal experimentation by complementing or potentially replacing them with new approach methodologies (NAMs), i.e., increasingly sophisticated in silico, in chemico, and in vitro approaches. We piloted development of one type of NAM, organ-on-chips (OOCs), in the highly challenging environment of a maximum (biosafety level 4) containment laboratory. Using a Risk Group 4 virus, Nipah virus (NiV), and two types of lung OOCs seeded with human or porcine cells, we demonstrated the recapitulation of key features of NiV lung infection, including viral infection, replication, and translocation, that are associated with proinflammatory cytokine secretion, immune cell recruitment, and disruption of the air-liquid interface barrier. We reproduced the known anti-NiV activity of remdesivir and evaluated that of another potential antiviral, zotatifin. Our results pave the way for similar applications of advanced microphysiological systems for modeling infections caused by high-consequence viruses.

Quantitative analysis of rodent behaviour is fundamental to neuroscience, preclinical drug discovery and neurotoxicology research. Although several commercial and open-source software packages are available for behavioural assessment, some are expensive, some require programming expertise, and some provide limited flexibility for user-defined experimental configurations. To address these limitations, we developed Robean, a freely available standalone software platform for automated rodent neurobehavioural analysis from both live camera feeds and pre-recorded videos. Robean provides an intuitive graphical user interface that enables users to design experimental arenas, define custom analysis zones, perform spatial calibration, and automatically track rodent movement without requiring programming knowledge. The software currently supports automated analysis of three widely used behavioural paradigms: the Morris Water Maze, Elevated Plus Maze, and Open Field Test. Robean extracts behavioural metrics including escape latency, path efficiency, platform crossings, target quadrant preference, thigmotaxis, locomotor activity, zone occupancy, arm entries, and centre exploration specific to behavioural tests. In addition, the software generates trajectory maps, occupancy heatmaps, comma-separated value (CSV) datasets, comprehensive PDF reports, and batch study summaries for multiple experimental sessions. Developed using open-source software technologies and distributed as a standalone freeware application, Robean provides an accessible and reproducible solution for behavioural neuroscience laboratories. Its modular architecture facilitates future integration of additional behavioural paradigms and analytical modules, making it a flexible platform for automated rodent behavioural assessment.

Understanding how -synuclein misfolding and spreading affect brain systems is central to synucleinopathy research, yet evidence from primate models remains limited. Here, we combined histology, awake structural MRI, awake resting-state fMRI, regional homogeneity, actimetry and touchscreen behavioral testing to longitudinally track the consequences of striatal -synuclein seeding in the common marmoset. Phosphorylated -synuclein inclusions were detected from 2 months post-injection and spread progressively to distributed cortical and subcortical regions bilaterally, accompanied by regionally specific structural atrophy. Functional imaging revealed widespread disruption of large-scale networks, particularly within fronto-limbic circuits, alongside reductions in local functional coherence. Despite the absence of overt motor or sleep deficits, animals exhibited a selective impairment in cognitive flexibility, with preserved learning and task engagement. These findings indicate that striatal -synuclein seeding induces a multi-scale reorganization of brain networks preferentially affecting circuits supporting cognitive flexibility and limbic processing, establishing a primate platform for studying early stages of synucleinopathy.

A major challenge in behavioral neuroscience is the lack of a unified software framework capable of implementing diverse paradigms across species and experimental setups. Researchers currently face a trade-off: they must either spend significant time developing custom, siloed solutions that hinder reproducibility, or incur substantial costs purchasing inflexible, closed systems. Here, we present Neurokraken, an open-source, Python-native platform designed to overcome these limitations. Neurokraken allows writing experiment progression entirely in standard python, while its core architecture automatically sets up a microcontroller for the connected hardware components and enables python side access with millisecond-precision timing and automatic logging. The system prioritizes ease of use and flexibility, enabling advanced series of events and conditions, the usage of python ecosystem code and packages within experiments, and the addition of any arduino-compatible electronic devices for custom experiments. As a result, users can easily create interactive virtual and real environments to engage, monitor, and record subjects. We present Neurokraken's versatility across a wide range of paradigms, for human and non-human primate psychophysics, and complex rodent behavior in both head-fixed and freely moving paradigms. Its modular design allows for rapid hardware reconfiguration, while a fully customizable user interface enables real-time monitoring and interactive experimental control without compromising timing precision. By uniting laboratory-grade precision with an accessible and flexible open-source philosophy, Neurokraken provides a single, powerful solution to design and execute next-generation behavioral experiments. We hope Neurokraken helps accelerate research, improve reproducibility throughout the neuroscience community, and make advanced behavioral experimentation more accessible through its substantial cost-efficiency.

Animal cell-cultured ingredients are entering the EU and UK pet food markets under frameworks that do not require pre-market, ingredient-level safety assessments, creating an ethical need for transparent safety disclosure. We present the first public safety dossier for this sector, describing the proprietary mouse embryonic stem cell line PE25 and its derived, non-viable cellular and conditioned media ingredient produced in food and feed-grade media. PE25 characterization confirmed Mus musculus identity, sterility, absence of mycoplasma and replication-competent retroviruses, and stable growth. Doxorubicin-induced p53 stress testing, CD44/BMI1 profiling, and soft agar assays showed no cancer-like traits and a non-tumorigenic profile; the final ingredient contains no viable cells. Independent OECD TG 471 and 487 assays confirmed non-genotoxicity. Heavy metals, biogenic amines, solvents, and chemical residues were below regulatory limits. Given process variability, we recommend case-by-case safety evaluation and propose this dossier as a model for responsible commercialization.

Dominance hierarchies are a fundamental aspect of social organization, enabling animals to minimize aggression and optimize access to resources. Previous studies have highlighted the energetic and physiological demands of dominant status, as well as the behavioral flexibility required of subordinates to navigate these hierarchies. Despite advancements in automated behavior tracking, limitations persist in tracking fine-scale, real-time interactions within complex social environments. Here, we developed and validated a novel RFID-based system to continuously monitor dominance hierarchies in group-housed male mice over 10 days. This system enabled unbiased behavioral inference across light phases and revealed spatial and temporal patterns of dominance behavior undetectable through traditional live-scored methods. Automated tracking accurately identified alpha individuals and consistently inferred linear hierarchies across cohorts, with greater precision for higher-ranked individuals. Behavioral metrics, such as transition frequencies and proximity to food zones, were consistent with dominance driven activity. Hormonal analyses revealed that higher-ranked mice exhibited increased leptin and peptide YY, consistent with heightened activity and satiety signaling, while lower C-peptide levels reflected greater metabolic demands of dominance. Furthermore, dominance rank was associated with differences in light-dark activity, which were in turn related to circulating hormone profiles. This study demonstrates the utility of automated RFID tracking in capturing dominance hierarchies with temporal and spatial granularity, while revealing links between social rank, metabolic regulation, and activity patterns advancing our understanding of social behavior dynamics.

In recent years, rodent touchscreen-based paradigms have gained popularity for their flexible task design, automated data collection and improved standardization. These devices minimize experimenter intervention and contribute to more replicable and reliable research. However, the high costs and proprietary hardware / software associated with commercially available systems present a financial barrier for smaller labs or researchers with limited resources. To address this issue, we developed "NC4Touch", an open-source scalable, modular rodent testing apparatus. It features three independent touchscreens and a feeding port for easy use in a wide array of tests comparable to those available with standard operant chambers, with the added benefit that visual stimuli can be highly customized. The system includes a user-friendly graphical interface that offers real-time control, task customization, video recording, and data management. We demonstrate its effectiveness in a visual discrimination task using both rats and mice. Several devices can be operated in parallel from the same computer and user interface, allowing high- throughput data collection. We provide detailed assembly instructions for the hardware and well- documented and easily-configurable software. We hope that the affordable open-source nature of NC4Touch will expand the scope of behavioral testing, allowing researchers to overcome traditional financial barriers and create a more collaborative community. Significance StatementTouchscreen-based testing is a powerful tool for assessing cognition in rodent models, however commercial systems remain cost-prohibitive for many researchers. NC4Touch is a rodent touchscreen apparatus that provides an open-source, customizable, and affordable alternative that enables high-throughput, cognitive testing in both mice and rats. By lowering financial barriers and promoting hackability, this system enables greater accessibility and collaboration in research.

The psychiatric utility of ketamine is limited by its dissociative and systemic side effects. Recently, to enable precision ketamine pharmacotherapy, we introduced SonoKet, ketamine-loaded acoustically activatable liposomes that enable focused ultrasound (FUS)-targeted ketamine delivery to millimeter-sized brain regions. In initial studies, we observed that SonoKet uncaging targeted ketamine to the ultrasound-treated brain region, while inducing greater electrophysiologic and behavioral functional effects than dose-matched free ketamine. To further define these uncaging-potentiated neuromodulatory effects, we used solid-phase microextraction (SPME) coupled to LC-MS/MS to investigate the effect of ultrasound and SonoKet uncaging on key neurotransmitters in real-time. SPME probes were used to sample ketamine, its metabolites, and glutamate, GABA, serotonin (5-HT), and dopamine in the medial prefrontal cortex (mPFC), nucleus accumbens (NAc), and retrosplenial cortex (RsC) of awake rats. Sampling occurred before and after intravenous administration of either SonoKet, free ketamine, or saline, with FUS targeted to either a frontolimbic or caudal brain region. FUS alone did not yield significant changes in neurotransmitter concentration, nor did it affect the pharmacodistribution of free ketamine. In contrast, FUS generally increased the neurotransmitter response to free ketamine, suggesting an ultrasonic potentiation of ketamine neuromodulation. SonoKet (0.75 mg/kg) uncaging with FUS elicited further elevations in glutamate, GABA, and 5-HT within the FUS-targeted region, along with an increase in dopamine in the NAc when the frontolimbic region was sonicated. These increases were similar to or higher than those induced by 10 mg/kg free ketamine alone or 0.75 mg/kg free ketamine combined with FUS, especially with frontolimbic SonoKet uncaging. Altogether, FUS potentiates ketamine-induced neuromodulation, with spatially specific and synergistically greater effects when ketamine is spatially localized via ultrasonic uncaging. This strategy could augment ketamine pharmacotherapy for psychiatric diseases, while limiting its dissociative and abuse liabilities. HighlightsO_LIFocused ultrasound potentiated ketamine-driven glutamate, serotonin, and dopamine release C_LIO_LILocalized ketamine delivery with SonoKet uncaging drove synergistically greater region-specific neurochemical responses C_LIO_LIUncaging boosts ketamine effects at a fraction of the ketamine dose C_LI Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=152 SRC="FIGDIR/small/741494v1_ufig1.gif" ALT="Figure 1"> View larger version (52K): org.highwire.dtl.DTLVardef@1745214org.highwire.dtl.DTLVardef@1b8e06borg.highwire.dtl.DTLVardef@95a840org.highwire.dtl.DTLVardef@15924ed_HPS_FORMAT_FIGEXP M_FIG C_FIG

With the rapid advancement of mRNA vaccines, lipid nanoparticles (LNPs) have emerged as pivotal carriers and adjuvants for non-mRNA vaccine modalities, driven by their superior nucleic acid delivery efficiency and intrinsic self-adjuvanting properties. In this study, we systematically evaluated various formulation strategies combining LNPs and the CpG adjuvant within a varicella-zoster virus glycoprotein E (VZV-gE) subunit vaccine framework. We demonstrated that uniform nanoparticles formed by LNP-encapsulated CpG (LNP-CpG), when simply admixed with the gE antigen, elicited superior immunogenicity compared to alternative encapsulation configurations. Intramuscular administration of a two-dose (LNP-CpG)+gE regimen significantly augmented both humoral and cellular immune responses in mice, markedly outperforming the commercial vaccine Shingrix (administered at a 1/10 human dose). Crucially, the identical regimen induced robust, comparable immune profiles to a full human dose of Shingrix in rhesus macaques. Furthermore, LNP-CpG displayed broad-spectrum utility across diverse vaccine platforms, demonstrating efficacy against both respiratory (RSV) and neurotropic (HSV) pathogens, compatibility with multiple modalities, including subunit (VZV-gE, RSV-Pre-F), live-attenuated (LA-HSV), and inactivated (i-HSV) vaccines; and versatile implementation in a combined VZV+RSV formulation. Collectively, our findings position LNP-CpG as a versatile, safe, highly efficacious adjuvant platform with substantial clinical translational potential, offering a compelling paradigm for next-generation vaccine development.

Effective animal monitoring is essential for assessing health, behavior, and environmental interactions, particularly in research and welfare contexts. This study presents a low-cost, open-source system designed for non-invasive monitoring of budgerigars (Melopsittacus undulatus), a small parrot species frequently used in animal behavior research. The system integrates a perch-based scale for voluntary weight measurement, a temperature sensor, and a camera for image capture, all controlled by a Raspberry Pi. By leveraging fine-tuned neural networks, the system achieves automated individual recognition with high accuracy, eliminating the need for invasive tagging methods. The modular design ensures accessibility, scalability, and minimal disturbance to the animals, while the accompanying software streamlines data collection, processing including labeling, and visualization. This approach provides a comprehensive solution for continuous monitoring, offering valuable insights for research and husbandry while prioritizing animal welfare.

Traditionally, complex behavioral tasks in mice have relied upon water restriction as an external motivator to increase task engagement. However, citric acid (CA) water, the technique whereby water is given ad libitum but made sour by the addition of CA, has emerged as an alternative to typical water restriction. Evidence suggests CA water can effectively motivate task performance while improving animal welfare in alignment with the 3Rs of animal research and reducing experimenter labor. While promising, the applicability of CA water in mice remains incompletely characterized with higher concentrations only tested in rats and "ramping" schedules, where mice progress to increasingly higher concentrations, indirectly examined. Here, we evaluate four CA concentration/schedule combinations for their effects on mouse health (weight changes, home cage behaviors, fecal counts) and motivation to drink regular water (lick counts, drinking behaviors) in female and male C57BL/6J mice. We find that a schedule ramping from 1% to 2% CA after one week is the easiest for mice to adapt to and sustained 2% CA use maintains robust lick counts for at least 5 weeks. Additionally, CA has been directly characterized for wheel-turning and touchscreen tasks, but not virtual reality (VR) tasks, an increasingly popular class of behavioral experiments. Therefore, we also assess how 2% CA affects motivation and task performance in two VR treadmill tasks (running and stopping task). We find that CA water does not improve task performance above that of mice given regular water, but does limit competing motivations and produce more uniform, reward-motivated behavior.