Oleic oil

Ciprofloxacin base (molecular weight: 331.34 Da), hyaluronic acid (LMW HA, MW 500 kDa), N-hydroxy succinimide (NHS), 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC),cremophore RH 40, tween 80, PEG 400, PEG 200, artificial mucin, oleic oil, linseed oil, caproyl 90, labrafil M, mineral oil, dulbecco’s modified eagle’s medium (DMEM), rhodamine, crystal violet, congo red, 4′,6-diamidino-2-phenylindole(DAPI), and fluorescein isothiocyanate (FITC) dye were purchased from Sigma Aldrich, Germany. Cystamine dihydrochloride was purchased from Avonchem, New Hampshire, UK) United Kingdom. All solvents used were of analytical grade.

To determine the capacity of EPA inclusion in enrichments to raise the n-3 LC-HUFA content in Artemia sp., five experimental enrichments (EPA 0–4) with increasing EPA content (0%–60%) were formulated (Table 1). The enrichments were prepared by mixing increasing amounts of a commercial oil (Incromega EPA 500TG; Croda, Barcelona, Spain) containing 63% EPA and 8% DHA in total fatty acids (TFA) with a second commercial oil (Oleic Oil; Sigma–Aldrich, Madrid, Spain) containing 77% oleic acid (OA, 18 : 1n-9) in TFA. Additionally, every enrichment preparation was mixed with constant amounts of soya lecithin (Korot SL, Alcoy, Spain) containing 54% linoleic acid (18 : 2n-6) and trace amounts of EPA and DHA as emulsifier. Also, enrichment preparations were fortified with 3 g kg⁻¹ vitamin E (DL-α-tocopherol acetate; Sigma–Aldrich, Madrid, Spain) and 2.5 g kg⁻¹ vitamin C (L-ascorbic acid, Asc; Sigma–Aldrich, Madrid, Spain) to prevent the oxidation according to Betancor et al. [24 (link)], Hamre et al. [13 (link)], and Roo et al. [10 (link)]. Once prepared, experimental enrichments were stored in a fridge at 4°C until used. During the feeding trial, three subsamples of each experimental enrichment (ca. 100 g), newly hatched Artemia sp. (ca. 100 g), and enriched Artemia sp. (ca. 100 g) were collected and analyzed for proximate and fatty acid composition (Table 2).