Bee Tracker-an open-source machine learning-based video analysis software for the assessment of nesting and foraging performance of cavity-nesting solitary bees.

The foraging and nesting performance of bees can provide important information on bee health and is of interest for risk and impact assessment of environmental stressors. While radiofrequency identification (RFID) technology is an efficient tool increasingly used for the collection of behavioral data in social bee species such as honeybees, behavioral studies on solitary bees still largely depend on direct observations, which is very time-consuming. Here, we present a novel automated methodological approach of individually and simultaneously tracking and analyzing foraging and nesting behavior of numerous cavity-nesting solitary bees. The approach consists of monitoring nesting units by video recording and automated analysis of videos by machine learning-based software. This software consists of four trained deep learning networks to detect bees that enter or leave their nest and to recognize individual IDs on the bees' thorax and the IDs of their nests according to their positions in the nesting unit. The software is able to identify each nest of each individual nesting bee, which permits to measure individual-based measures of reproductive success. Moreover, the software quantifies the number of cavities a female enters until it finds its nest as a proxy of nest recognition, and it provides information on the number and duration of foraging trips. By training the software on 8 videos recording 24 nesting females per video, the software achieved a precision of 96% correct measurements of these parameters. The software could be adapted to various experimental setups by training it according to a set of videos. The presented method allows to efficiently collect large amounts of data on cavity-nesting solitary bee species and represents a promising new tool for the monitoring and assessment of behavior and reproductive success under laboratory, semi-field, and field conditions.