Journal Article

Integrative computational modeling framework linking mycotoxin contamination, microbial hazards, and antimicrobial resistance risk in dairy systems

Khan, Sumbal, Pari, Bakht, Khan, Ruby

2025

Overview

Background Milk can sometimes carry harmful mycotoxins, which pose health risks, especially in hot and humid areas. We collected samples from both farms and households in Khyber Pakhtunkhwa (KP), Pakistan, to see how common these toxins are, whether the genes that make them are present, and which environmental factors might make contamination worse. Methods We tested raw milk for aflatoxins (AFM1, AFM2), ochratoxin A (OTA), and zearalenone (ZEN) using TLC, HPLC, and UHPLC-MS/MS. All testing procedures were validated following ICH Q2(R2), ISO 17025, and FDA guidance. PCR was applied to check for the genes aflC , otaA , and zen1 . We then combined chemical, molecular, microbial, and environmental data and used multivariate statistics and PLS-DA modeling to find the main factors driving contamination. Results Milk from farms had higher mycotoxin levels than household milk. Average AFM1 concentrations were g/kg in farm milk and g/kg in domestic milk (p < 0.001). AFM2 and OTA showed similar patterns, while ZEN was below detection in all samples. UHPLC-MS/MS confirmed the HPLC findings and offered greater sensitivity. The genes aflC and otaA were found in 68% of farm samples and were strongly linked to AFM1 and OTA levels, while zen1 was absent. High temperature (over 28°C) and humidity (over 75%) were associated with increased contamination. PLS-DA modeling effectively distinguished high- and low-risk samples (AUC = 0.92), highlighting AFM1 concentration, aflC presence, and humidity as key predictors. Conclusions Combining chemical testing, gene screening, and environmental monitoring provides a practical way to detect and evaluate mycotoxin risk in milk. Farm milk showed higher contamination than household milk, emphasizing the need for targeted monitoring and preventive measures. Identifying environmental thresholds and risk factors can support early interventions to improve food safety and protect public health.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Aflatoxins / Aflatoxins - analysis / AFM1 / Analysis / Animals / Antibiotics / Antimicrobial agents / Antimicrobial resistance / Bioinformatics / Biological Microscopy / Biomedical and Life Sciences / Biosynthetic genes / Cattle / Chromatography / Chromatography, High Pressure Liquid / Computational biology / Computer Simulation / Contamination / Cross-sectional studies / Dairy products / Dairying / Drug resistance / Drug resistance in microorganisms / Environmental factors / Environmental monitoring / Farms / Food contamination / Food contamination & poisoning / Food Contamination - analysis / Food safety / Genes / Health risks / Heat resistance / High performance liquid chromatography / High temperature / Households / Humid areas / Humidity / Life Sciences / Metabolites / Methods / Microbial contamination / Microbiology / Microorganisms / Milk / Milk - chemistry / Milk - microbiology / Milk contamination / Modelling / Monsoons / Multivariate analysis / Mycology / Mycotoxins / Mycotoxins - analysis / Ochratoxin A / Ochratoxins - analysis / OTA / Pakistan / Parasitology / Prevention / Public health / Risk factors / Surveillance / Tandem Mass Spectrometry / Test procedures / Testing procedures / Toxicology / Toxins / UHPLC-MS/MS / Virology / Zearalenone