4.5. Eco-Toxicological Profile: Allelopathy and Artemia salina (brine shrimp) Lethality Assays

Allelopathy bioassay was carried on the seeds of the commercial lettuce variety Lollo bionda because of its fast germination rate and high sensitivity. The detailed procedure was conducted as previously reported [10]. Seeds were treated with scalar P. mahaleb extract concentrations (0.1–20 mg/mL) and considered germinated for observed root length ≥ 1 mm, after the third day of treatment. Artemia salina cysts were cultivated in oxygenated artificial sea water (1 g cysts/L). After 24 h, brine shrimp larvae were gently transferred with a pipette in 6 well plates containing 2 mL of the extract at different concentrations (0.1–20 mg/mL) in artificial sea water. The detailed protocol is reported in our previous paper [10].