Detection of infertile eggs prior to incubation can lead to an increase in the hatchability rate and prevent the wastage of billions of non-fertile eggs ended up by failed incubation. In this study, the feasibility of a line-scan hyperspectral imaging system in the visible and short-wavelength near-infrared region was assessed for early detection of non-fertile eggs on day 0 before incubation. A total of 227 white-shell eggs including 131 fertile and 96 infertile eggs were collected from a flock with similar conditions in terms of hen age, feeding, and management. Hyperspectral images of eggs were captured on day 0 before incubation in a transmittance mode of illumination and then the eggs were incubated in a commercial incubator. The edge detection method was used to segment the egg, including both the white and yolk, from the background, and the image spectral information was extracted from the egg region. After applying various pretreatment methods, different classifiers including soft independent modeling of class analogy (SIMCA), linear discriminant analysis (LDA), quadratic discriminant analysis (QDA), and artificial neural networks (ANN) classifiers were utilized to extract the predictive models. Following the acceptable results of SIMCA analysis accomplished by 1st derivative pretreatment (accuracy of 86.67%), the discrimination power plot was used to select the most informative wavebands. The results showed that by using fewer variables in effective wavebands better performance (precision and accuracy of 92.59% and 93.33%, respectively) could be obtained in comparison with the ANN classifier based on the whole spectral data (precision and accuracy of 89.29% and 91.11%, respectively). This study revealed the potential application of hyperspectral transmittance imaging in the Vis-SWNIR region to discern the fertile and infertile eggs before starting the incubation process.© 2024. The Author(s).