Bifenthrin

All the used chemicals (pure and of analytical grade) were obtained from the stores of the National Research Centre, Egypt. Bifenthrin (purity 98%) was purchased from Sigma, MO, USA. Flowers of E. purpurea were obtained from IMTENAN Company (Production and sale of nutritional and healthy products, Giza, Egypt); the company collected and dried the flowers in accordance with relevant institutional, national, and international guidelines and legislation. The plant was examined by botanical specialists (Faculty of pharmacy, Ain-Shams university, Cairo, Egypt) and was found carrying the taxonomic serial number 3728.

Acaricides (96% cyflumetofen, CAS: 400882-07-7; and 97.2% bifenthrin CAS: 82657-04-3) were purchased from Sigma-Aldrich Co. (Saint Louis, USA). ELISA kits were purchased from Jiangsu Boshen Biotechnology Co., Ltd. (Nanjing, Jiangsu, China). The Total RNA Kit II was purchased from Omega (Seoul, Korea).

Bifenthrin (purity; 99.0%), butachlor (95.9%) standards, and ammonium formate (10 M) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Commercial pesticide products were Talstar [Bifenthrin 1 EC (NongHyup Chemical Co., Ltd., Seongnam, Korea)] and Macet [butachlor 5 GR (FarmHannong Co., Ltd., Seoul, Korea)]. HPLC grade acetonitrile (MeCN) and ethyl acetate (EA) were obtained from Samchun Chemical (Seoul, Korea) and from Duksan Pure Chemical (Seoul, Korea), respectively. LC–MS grade methanol (MeOH) and formic acid (>99%) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). LC–MS grade water was obtained from Merck (Darmstadt, Germany). Acetic acid (99.5%) was purchased from Junsei Chemical Co., Ltd. (Tokyo, Japan). QuEChERS extraction original kit (4 g of magnesium sulfate; MgSO₄ and 1g of sodium chloride; NaCl), EN 15662 kit (4 g of MgSO₄, 1 g of NaCl, 1 g of sodium citrate; Na₃Citr·2H₂O, and 0.5 g of sodium hydrogen citrate sesquihydrate; Na₂HCitr·1.5H₂O), d-SPE, PSA (25 mg of PSA and 150 mg of MgSO₄), and GCB (25 mg of PSA, 7.5 mg of bulk carbonate, and 150 mg of MgSO₄) were obtained from Agilent Technologies (Santa Clara, CA, USA). Oasis PRiME HLB cartridge plus light (100 mg) was purchased from Waters Corporation (Milford, MA, USA).

P. synxantha PS1 was isolated from oil well-produced water (Shengli Oilfield, Shandong Province, China). E. coli DH5α (Invitrogen, USA) and plasmid pMD19-T (TaKaRa, Japan) were used for gene cloning and sequencing of AT-containing and AT-truncated carboxylesterases. Plasmid pET-28a (Novagen, USA) was used as a vector to construct the protein expression plasmid in E. coli BL21 (DE3). The substrates fenpropathrin, cypermethrin, fenvalerate, bifenthrin, and p-nitrophenyl esters were purchased from Sigma (St. Louis, MO, USA). All other chemicals were of analytical grade and purchased from local markets.

D-cyphenothrin (98% purity) was obtained from Wuhan Yuancheng Pharm Co., Ltd., China. Technical-grade tetramethrin (99%), permethrin (97%), bifenthrin (96%), allethrin (93%), and chlorempenthrin (94%) were purchased from Sigma-Aldrich, USA. Chromatographic grade acetone and acetonitrile was obtained from Fisher Scientific, USA. All other chemicals and solvents used in this study were of analytical grade. Stock solutions were prepared by dissolving SPs in chromatographic grade acetone at a concentration of 10 g·L⁻¹ and stored in dark bottles at 4 °C.
Luria-Bertani medium (LB) was composed of 10 g tryptone; 5 g yeast extract; and 10 g NaCl dissolved in 1-L distilled water. Mineral salt medium (MSM) (g·L⁻¹) containing (NH₄)₂SO₄, 2; Na₂HPO₄·12H₂O, 1.5; KH₂PO₄, 1.5; MgSO₄·7H₂O, 0.2; CaCl₂·2H₂O, 0.01; and FeSO₄·7H₂O, 0.001 was used for biodegradation assays whereas solid medium was additionally added with 1.5% agar. The pH of both culture media was adjusted to 7.0 and autoclaved at 121 °C for 20 min.

D-phenothrin (98% purity) was obtained from Jiangsu Yangnong Chemical Group Co., Ltd., China. Technical-grade permethrin, cyhalothrin, β-cypermethrin, deltamethrin, fenpropathrin, and bifenthrin were purchased from Sigma–Aldrich, United States and high-performance liquid chromatography (HPLC)-grade acetonitrile was purchased from Fisher Scientific, United States. All other chemicals and solvents were of analytical grade. SPs were dissolved in dimethyl sulfoxide (DMSO) or acetone at a stock concentration of 10 g⋅L^-1, and stored in dark bottles at 4°C.
Mineral salt medium (MSM) containing (g⋅L^-1) (NH₄)₂SO₄, 2; MgSO₄⋅7H₂O, 0.2; CaCl₂⋅2H₂O, 0.01; FeSO₄⋅7H₂O, 0.001; Na₂HPO₄⋅12H₂O, 1.5; and KH₂PO₄, 1.5; and Luria-Bertani (LB) medium containing (g⋅L^-1) tryptone, 10; yeast extract, 5; and NaCl, 10 were used in this study. Both culture media were adjusted to pH 7.3 and sterilized at 121°C for 20 min.

Seven analytical standard pyrethroid insecticides (bifenthrin, cypermethrin, deltamethrin, esfenvalerate, permethrin, lambda-cyhalothrin, and fenvalerate) with 99.1, 99.0, 99.9, 98.5, 91.8, 98.0, and 97.0% purity, respectively (Table S1 Supplementary Material) were purchased from Sigma-Aldrich (Steinheim, Germany). Acetonitrile, methanol, formic acid, and acetone were chromatographic grade and purchased from Merck (Darmstadt, Germany). Other chemicals of magnesium sulphate (MgSO4: 98% purity), sodium acetate (NaAc; 99% purity), primary secondary amine (PSA: 99% purity), sodium chloride (NaCl: 99% purity), and ammonium formate (98% purity) were purchased from Sigma-Aldrich (Steinheim, Germany).