Molecular Remodeling of Milk Fat Globules Induced by Centrifugation: Insights from Deep Learning-Based Detection of...

Milk adulteration through centrifugation, which artificially reduces the somatic cell count (SCC), represents a significant challenge to food authenticity and public health. This fraudulent practice alters the native molecular architecture of milk, masking inflammatory conditions such as subclinical mastitis and distorting product quality. Conventional analytical and microscopic techniques remain insufficiently sensitive to detect the subtle physicochemical changes associated with centrifugation, highlighting the need for molecular-level, data-driven diagnostics. The dataset included 128 paired raw milk samples and approximately 25,000 bright-field micrographs acquired across multiple microscopes, of which 95% were confirmed to be of high quality. In this study, advanced machine learning (ML) and deep learning (DL) approaches were applied to identify centrifugation-induced alterations in raw milk microstructure. Bright-field micrographs (pixel size 0.27 µm) of paired unprocessed and centrifuged samples were obtained under standardized optical conditions and analyzed using convolutional neural networks (ResNet-18/50, Inception-v3, Xception, NasNet-Mobile) and hybrid attention architectures (MaxViT, CoAtNet). Model performance was evaluated using the harmonic average of recalls across five micrographs per sample (HAR₅). Human microscopy experts (n = 4) achieved only 18% classification accuracy-below the random baseline (25%)-confirming that centrifugation-induced modifications are not visually discernible. In contrast, DL architectures reached up to 97% accuracy (HAR₅, Xception), successfully identifying subtle molecular cues. Class activation and sensitivity analyses indicated that models focused not on milk fat globule (MFG) boundaries but on high-frequency nanoscale variations related to the reorganization of casein micelles and solid non-fat fractions. The findings strongly suggest that centrifugation adulteration constitutes a molecular reorganization event rather than a morphological alteration. The integration of optical microscopy with AI-driven molecular analytics establishes deep learning as a precise and objective tool for detecting fraudulent milk processing and improving food integrity diagnostics.