We are developing a novel detection method for Varroa mites in Honey Bee hives. Nearly 50% of beehives are lost yearly by colony collapse disorder (CCD), a crisis exacerbated by Varroa mites. Traditional methods for detecting these mites, like the powdered sugar test and alcohol test, are disruptive to the hive and not very accurate. To mitigate this, our iGEM project introduces a biosensor-based testing method targeting the guanine in mite excrement, a significant component. When detected, it triggers a genetic pathway leading to the production of mScarlet-I, a protein measurable through visual analysis or a spectrophotometer, offering a more accurate and less intrusive mite detection process. This innovation is particularly significant for North Dakota, the U.S.' top honey producer, where agriculture and beekeeping are vital. Our biosensor not only aids in addressing CCD but also aligns with the state’s shift towards integrating autonomous systems and Agtech, promising a substantial impact on the local agricultural ecosystem.