Aflatoxin B₁ Metabolism of Reared Alphitobius diaperinus in Different Life-Stages

The presence of carcinogenic aflatoxins in food and feed is a major issue. In prior studies, aflatoxin B₁ (AfB₁) and known primary metabolites were absent from Lesser Mealworm (LMW, Alphitobius diaperinus) reared on contaminated diets. LMW is a promising alternative protein source. The objectives of this stu\dy were to determine whether LMW can be reared on AfB₁-contaminated feed in each life-stage, and to gather more insight into potential metabolites formed. Results suggested no adverse effects in terms of survival/growth when three stages of LMW (larvae, pre-pupae, beetles) were exposed to feed containing AfB₁ concentrations of 200 and 600 µg/kg for 48 h. Insect and frass samples were analyzed by LC-MS/MS and high-resolution MS to, respectively, quantify concentrations of AfB₁ and its major metabolites, and determine secondary metabolites. No AfB₁ or major metabolites were quantified in the insect samples. Mass balance calculations showed that up to 40% of spiked AfB₁ could be recovered in the frass, in the form of AfB₁, aflatoxicol and AfM₁. HRMS results suggested the presence of additional metabolites in the frass, but, due to lack of commercially available reference standards for these compounds, exact identification and quantification was not possible. More research is needed to verify the absence of toxicity.