A simple and low-cost artificial blood feeding system for mosquito colony maintenance and experimental applications.

Reliable and consistent blood feeding is crucial for maintaining mosquito colonies in the laboratory and for supporting scientific research, including vector competence studies (i.e., oral inoculation). However, artificial blood feeding systems often require costly equipment (e.g., Hemotek), and the use of live animals as hosts raises ethical concerns, limiting their practicality. In this study, we developed a simple, low-cost blood feeding system consisting of a perforated plastic pouch heated with a disposable hand warmer and evaluated its effectiveness in supporting blood feeding of 3 mosquito species: Culex quinquefasciatus Say, Aedes aegypti L., and Aedes triseriatus Say. Specifically, we compared feeding success between fresh and previously frozen bovine blood across species. Our results showed that lab-reared Cx. quinquefasciatus achieved the highest engorgement rates (up to 95.7%) across all treatments, whereas field-collected Cx. quinquefasciatus failed to feed on any blood source. Aedes aegypti and Ae. triseriatus exhibited significantly higher feeding success on fresh blood compared to previously frozen blood, with 2.1-fold and 9.7-fold increases, respectively. The perforated pouch system supported full engorgement with minimal leakage and did not require electricity or specialized equipment. These results support the utility of this method for colony maintenance of some lab-adapted mosquito species and vector competence experiments, particularly in settings with limited access to vertebrates or other blood-feeding systems.