A hyperspectral-multispectral line-scan imaging system was developed for differentiation of wholesome and systemically diseased chickens. In-plant testing was conducted for chickens on a commercial evisceration line moving at a speed of 70 birds per minute. Hyperspectral image data was acquired for a calibration data set of 543 wholesome and 64 systemically diseased birds and for a testing data set of 381 wholesome and 100 systemically diseased birds. The calibration data set was used to develop the parameters of the imaging system for conducting multispectral inspection based on fuzzy logic detection algorithms using selected key wavelengths. Using a threshold of 0.4 for fuzzy output decision values, multispectral classification was able to achieve 90.6% accuracy for wholesome birds and 93.8% accuracy for systemically diseased birds in the calibration data set and 97.6% accuracy for wholesome birds and 96.0% accuracy for systemically diseased birds in the testing data set. By adjusting the classification threshold, 100% accuracy was achieved for systemically diseased birds with a decrease in accuracy for wholesome birds to 88.7%. This adjustment shows that the system can be feasibly adapted as needed for implementation for specific purposes, such as paw harvesting operations or prescreening for food safety inspection. This line-scan imaging system is ideal for directly implementing multispectral classification methods developed from hyperspectral image analysis.