Recycling deteriorated silage to remove hazardous mycotoxins and produce a value-added product.

Silage, an important forage feed, contains hazardous mycotoxins due to spoilage caused by unreasonable management. Deteriorated silage becomes a mycotoxin source and threatens human health and the eco-environment. Recycling deteriorated silage and exploiting beneficial substances would be profitable and environmentally friendly. Squalene [60.3-73.9 mg/kg fresh matter (FM)] and 6 types of mycotoxins (4.56-10,080 ug/kg FM) were found in deteriorated silages. To clarify the source and synthesis mechanism of squalene, alfalfa was ensiled at low temperature (LT, 3-20 ℃), 25 ℃ (T25), 30 ℃ (T30) or 35 ℃ (T35) for 10, 40 and 70 d. The highest squalene was detected when alfalfa ensiled for 40 d (P = 0.033) or ensiled at LT and T30 (P < 0.001). Squalene source was traced as lactic acid bacteria (LAB) using next-generation sequencing. Multiple linear regression models inferred that squalene synthase of LAB positively contributed to the squalene synthesis but was negatively adjusted by ammonia-N during ensiling. Two promising squalene-producing LAB strains were screened from alfalfa silage, which fermented deteriorated silage to enhanced squalene yield (190~279 mg/L) with low cost and high mycotoxin removal ratios (up to 85.5%). Therefore, the environmentally friendly strategy of recycling deteriorated silage to produce beneficial squalene was created.