Co-occurrence of mycotoxins in poultry feed from Lahore and Rawalpindi divisions of Punjab, Pakistan.

Mycotoxin contamination of poultry feed remains a significant food safety concern due to the frequent co-occurrence of multiple mycotoxins that may cause additive or synergistic effects. This study investigated the occurrence and co-occurrence of aflatoxins (AFs), ochratoxin A (OTA) and fumonisins (FBs) in poultry feed from two major regions of Punjab, Pakistan. A total of 280 feed samples from Lahore and Rawalpindi divisions were screened using thin-layer chromatography and quantified by ELISA, while positive samples were confirmed by LC-ESI+-MS. Overall, 77.9% of the samples were contaminated with at least one mycotoxin, with prevalence rates of 79.3% in Lahore and 76.4% in Rawalpindi. Co-occurrence of two or more mycotoxins was observed in 42.1% of the positive samples, most commonly aflatoxins with fumonisins. Measured levels ranged from 4.16 to 251 µg/kg for AFs, 1.9 to 509 µg/kg for OTA and 210 to 49,200 µg/kg for FBs, frequently exceeding maximum limits.