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4 protocols using chromic oxide

1

Biofilm Formation Inhibition Assay

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All of the solutions were prepared in double-distilled water. Oxalic acid, phosphoric acid (85%), perchloric acid (70%), hydrochloric acid (37%), nitric acid (63%), sulfuric acid, hydrogen peroxide (30%), ethanol (95.5%), methanol (95%), acetone, chromic oxide, sodium hydroxyl, copper chloride (II), brilliant blue G, glycerol, and dimethyl sulfoxide were provided by EMD Millipore (Billerica, MA, USA). Crystal violet and aluminum foil (99.99%) were bought respectively from Cib Biotech Co (Tehran, Iran) and Kingcheng (Shanghai, China). NaH2PO4, Na2HPO4, BSA, and MTT reagents were provided by Sigma-Aldrich Co. (St Louis, MO, USA). MWCNT (outer diameter: 8–15 nm) was bought from CheapTubes.com (Cambridgeport, VT, USA). Two strains of bacteria containing E. coli ATCC: 25922 and S. aureus ATCC: 25923 as biofilm former were provided by Pasteur Institute of Iran (Tehran, Iran).
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2

Antimicrobial Efficacy of Nanomaterials

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Crystal violet and aluminum foil (99.99% Al) were bought from Cib biotech Co (Tehran, Iran) and Kingcheng (Beijing, China), respectively. Oxalic acid, phosphoric acid (85%), perchloric acid (70%), hydrochloric acid (37%), nitric acid (63%), sulfuric acid, hydrogen peroxide (30%), ethanol (95.5%), methanol (95%), acetone, chromic oxide, sodium hydroxyl, copper chloride (II), Coomassie Brilliant Blue G-250, and dimethyl sulfoxide were purchased from Merck Co. (Darmstadt, Germany). NaH2PO4, Na2HPO4, BSA, MTT reagent, enzymes, Hank’s Balanced Salt Solution (HBSS), alpha-minimal essential medium (αMEM), and FBS were bought from Sigma Aldrich (Darmstadt, Germany). Multi-walled CNTs (MWCNTs) were obtained from CheapTubes (Cambridgeport, VT, USA). FA was obtained from Alfa Aesar (Ward Hill, MA, USA) and studied. Presto Blue (PB) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Two strains of bacteria containing Escherichia coli (ATCC: 25922) and Staphylococcus aureus (ATCC: 25923) were provided by Pasteur Institute (Tehran, Iran) for biofilm formation.
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3

Microalgal Oil and Lipid Bioaccessibility

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Microalgal oil extracted from Schizochytrium sp. was purchased from Source-Omega LLC. (Chapel Hill, NC, USA). Fish oil extracted from menhaden was purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA), and soybean oil was purchased from Ottogi (Pyeongtaek, Gyeonggi-do, Korea). Bile salts, α-amylase, lipase from porcine pancreas, bovine serum albumin, pancreatin from porcine pancreas, mucin from porcine stomach, sodium molybdate, chromic oxide, and potassium dichromate were purchased from Sigma-Aldrich Chemical Co. Triundecanoin, a standard for fatty acid analysis, was purchased from NU-CHEK PREP, Inc. (Elysian, MN, USA) and Supelco 37 Component FAME Mix was purchased from Supelco Inc. (Bellefonte, PA, USA).
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4

Formulating Iso-Energetic Diets for Rainbow Trout Nutrition

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Two iso-energetic, iso-nitrogenous and iso-lipidic experimental diets were formulated to meet the nutrient requirements of rainbow trout [20 ]. Diets differed according to the added lipid source: the control diet (FO) was formulated with cod liver oil whereas the plant-derived diet (LO) was based on linseed oil. The detailed formulations and the proximate and fatty acid compositions are shown in Tables 1 and 2, respectively. The FO diet was particularly rich in n-3 LC-PUFA in which EPA (6.6 mg/g of dry matter (DM)) and DHA (10 mg/g DM) were the major fatty acids. In contrast, the LO diet contained mainly ALA (38.6 mg/g DM) and residual levels of n-3 LC-PUFA (1.2 mg/g DM). Chromic oxide (Sigma-Aldrich) was added at 10 g/kg DM to each experimental diets intended for the digestibility trial in order to serve as indigestible marker. The experimental diets were produced as previously described [42 (link)]. Briefly, the dry components were homogenised (SM 20, Guangzhou Both-Win, Guangdong, China) before and after the oil addition and then after the water addition. After cold extrusion (HI 2251, Simplex, Paris, France) and freeze-drying, the diets were stored at -20°C until feeding or analysis.
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