Attomolar-sensitive milk fever sensor using 3D-printed multiplex sensing structures

The diagnosis of milk fever or hypocalcemia in lactating cows has a significant economic impact on the dairy industry. It is challenging to identify asymptomatic subclinical hypocalcemia (SCH) in transition dairy cows. Monitoring subclinical hypocalcemia in milk samples can expedite treatment and improve the health, productivity, and welfare of dairy cows. In this study, an attomolar-sensitive sensor is developed using extrusion-based 3D-printed sensing structures to detect the ratio of ionized calcium to phosphate levels in milk samples. The unique geometries of the lateral structure of 3D-printed sensors, along with the wrinkled surfaces, provide a limit of detection down to the attomole (138 aM) concentration of the target analyte. The calcium-to-phosphate ratio in milk samples not only provides early disease indications but also enables on-site testing. This highly selective test is validated using real milk and blood samples, and the results are compared with those of commercial meters. This fast response (~10 s) low-cost sensor opens a promising tool for the farm-side diagnostic of dairy cows that can promote best practice management of dairy cows.