Abstract:
Automated behavioural tracking tools are increasingly valuable for reducing observer bias,
improving reproducibility, and enabling high-throughput data collection in animal behaviour
research. Recent developments in open-source and customizable tracking platforms have
enhanced accessibility and analytical efficiency across species, while modern programming
tools like R and Python enable affordable and precise behavioural quantification, particularly
valuable for research in resource-limited settings. Existing commercial and open-source
packages provide useful frameworks, yet they are often constrained by limited flexibility,
high costs, or inadequate adaptability to species-specific behaviours, particularly in non-
model organisms such as amphibian larvae. To address this gap, we developed a custom
Python-based tracking script capable of quantifying locomotor activity from standard video
recordings The tool automates the detection and tracking of individual larvae, extracting
continuous X–Y positional data and generating visual outputs such as trajectory plots and
heat maps to represent spatial activity patterns. Output files are produced in Excel-
compatible format, allowing seamless integration with statistical workflows. Behavioral
trials were conducted to evaluate both short-term repeatability and developmental
consistency of locomotor traits in tadpoles, demonstrating the script’s ability to capture fine-
scale variation in activity levels over time. Preliminary validation against manual scoring
confirmed high accuracy and reliability, highlighting its potential as a robust alternative to
observer-based measurements. Importantly, the script’s open and adaptable structure allows
researchers to modify tracking parameters according to study needs, extending its
applicability beyond amphibians to other small aquatic organisms. This custom tool
therefore offers a flexible, low-cost, and reproducible approach for behavioural ecologists
and conservation biologists investigating movement and activity in larval stages. By
facilitating detailed and standardized quantification of locomotion, it contributes to
advancing questions in developmental biology, personality research, and ecological
assessments of amphibian populations—areas of growing significance in light of global
amphibian declines.
