Fungal Contamination of Dairy Feed and Major Mycotoxin Transfer: A Risk Evaluation for Animal Exposure and Health

This study was focused on the assessment of fungal occurrence, mycotoxin dynamics, aflatoxin carry-over, and associated biochemical responses in dairy cattle. Moisture emerged as the dominant factor for fungal communities, promoting the co-proliferation of fungal genera adapted to high water activity conditions (a_w > 0.90) and antagonism against xerotolerant and xerophilic species. <i>Aspergillus</i> spp. dominated dry substrates (a_w < 0.75), <i>Fusarium</i> spp. showed strong positive associations with high-moisture matrices (a_w > 0.90), and <i>Penicillium</i> spp. exhibited intermediate, substrate-dependent behavior. Mycotoxin levels fluctuated non-linearly, independently of fungal counts: ochratoxin A (OTA) concentrations in corn silage increased from approximately 12 μg/kg at the onset of the ensiling period to >240 μg/kg at silo opening, indicating dynamic mycotoxin accumulation during storage, while zearalenone (ZEA) oscillated from 40 to 170 µg/kg. Despite the variation in total aflatoxins (AFLA-T) across feed matrices, aflatoxin M1 (AFM1) in milk remained low (0.0020-0.0093 μg/kg), confirming limited carry-over. Serum biochemical parameters-alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), total bilirubin (BIL-T), total protein (PROT-T)-remained within physiological limits, yet multivariate analyses revealed metabolic modulation linked to aflatoxin exposure. AFM1 explained >7% of the variance in serum biochemical profiles according to PERMANOVA (<i>p</i> = 0.002), showed significant MANOVA effect (Pillai = 0.198), and displayed a significant canonical association (<i>p</i> < 10^-13). Linear discriminant analysis further separated Normal vs. Borderline hepatic profiles, indicating subclinical physiological adaptation to chronic low-dose exposure.