Technical Abstract: Recently, hyperspectral image analysis has proved successful for a target detection problem encountered in remote sensing as well as near sensing utilizing in situ instrumentation. The conventional global bi-level thresholding for target detection, such as the clustering-based Otsu’s method, has been inadequate for the detection of biologically harmful material on foods that has a large degree of variability in size, location, color, shape, texture, and occurrence time. This paper presents multistep-like thresholding based on kernel density estimation for the real-time detection of harmful contaminants on a food product presented in multispectral images. We are particularly concerned with the detection of fecal contaminants on poultry carcasses in real-time. In the past, we identified 2 optimal wavelength bands and developed a real-time multispectral imaging system using a common aperture camera and a globally optimized thresholding method from a ratio of the optimal bands. This work extends our previous study by introducing a new decision rule to detect fecal contaminants on a single bird level. The underlying idea is to search for statistical separability along the two directions defined by the global optimal threshold vector and its orthogonal vector. Experimental results with real birds and fecal samples in different amounts are provided.