The ascomycete Fusarium equiseti K3 was isolated from HCH-contaminated soil collected from a former obsolete pesticide store in Kitengela, Kenya (GPS: 01.49 S, 37.048E) and its draft genome sequence was described earlier [36 (link)]. The fungus was not found to degrade HCH (see below). Fungal pre-cultures were cultivated at 25 °C on potato dextrose agar (PDA; BD Difco, USA) prior to the transfer to 13C glucose/glucose-containing minimum salt medium (MSM) agar. The HCH degrading bacterium Sphingobium sp. S8 was isolated from the same soil as the fungus and its draft genome sequence was described by Khan et al [37 (link)]. The bacterium was cultivated on Luria-Bertani (LB) (DSMZ medium 381) agar medium treated with γ-HCH (97%; Sigma) to a final concentration of 100 µg mL−1. To prepare the inoculum of the S8 strain, a 72-h old culture (OD600 ~1.5) of strain S8 in LB medium was centrifuged at 12,000 × g at 15 °C for 10 min, the cell pellet washed twice with MSM medium, and resuspended in the same medium to a concentration of 7.2 × 108 cells mL−1.
γ hch
Manufactured by Merck Group
Sourced in Germany
γ-HCH is a laboratory equipment product. It is used for analytical purposes.
Automatically generated - may contain errors
Lab products found in correlation
α hch, by Merck Group (2 mentions)
β hch, by Merck Group (1 mentions)
Salmon sperm dna, by Merck Group (1 mentions)
Dnase 1, by Takara Bio (1 mentions)
Endrin, by Merck Group (1 mentions)
Methoxychlor, by Merck Group (1 mentions)
Chlordane, by Merck Group (1 mentions)
Heptachlor epoxide, by Merck Group (1 mentions)
Heptachlor, by Merck Group (1 mentions)
Aldrin, by Merck Group (1 mentions)
Petroleum ether, by Merck Group (1 mentions)
Pp dde, by Merck Group (1 mentions)
N hexane, by Merck Group (1 mentions)
Diethyl ether, by Merck Group (1 mentions)
Malathion, by Merck Group (1 mentions)
Zinc chloride, by Loba Chemie (1 mentions)
Potassium dichromate, by Loba Chemie (1 mentions)
Sodium hydroxide (naoh), by Loba Chemie (1 mentions)
Lindane γ hch, by Merck Group (1 mentions)
1 pentanol, by Merck Group (1 mentions)
6 protocols using γ hch
1
Fusarium equiseti K3 Genomic Profiling
2
Investigating Persistent Organic Pollutants
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Salmon sperm DNA with an average molar mass of 1.3 × 106 Da and %G-C content of 41.2%, α-HCH, β-HCH, and γ-HCH were purchased from Sigma Chemical Co. (St. Louis, MO, USA). DNase I and DNase I buffer (10 × ) were purchased from Takara Bio Company (Dalian, Liaoning, China). The physicochemical properties of α-HCH, β-HCH, and γ-HCH are given in Supplementary Table 4 . All chemical reagents were of analytical grade and used without further purification. DNA stock solution of 1 g L−1 was prepared by dissolving 0.1 g DNA in 100 mL Tris-HCl buffer (10 mmol L−1, pH 7.0). Stock solutions of α-HCH, β-HCH, or γ-HCH (1 g L−1) were prepared using methanol as solvent. HCH working solutions were prepared by further diluting their stock solution by methanol to 10 mg L−1. All solutions were stored at 4 °C in a refrigerator before use.
3
HCH Isomers Quantification Protocol
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Hexane (LiChrosolv grade), Acetone (99.8%), and γ-HCH (97%) were either purchased from Sigma (Munich, Germany) or Merck (Darmstadt, Germany) and were of analytical grade. 13C-glucose (D-glucose13C6, 99%) was obtained from Cambridge Isotope Laboratories, Inc, (USA) The analytical standard containing a mixture of the HCH isomers, α: β: γ: δ = 1:1:1:1 was obtained from Merck (Darmstadt, Germany).
4
Extraction and Analysis of Persistent Organic Pollutants
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Standard OCPs including pp-DDT, pp-DDD, pp-DDE, α HCH, γ HCH, heptachlor, heptachlor epoxide, aldrin, endrin, chlordane, methoxychlor, and HCB were obtained from Sigma–Aldrich (Germany). Petroleum ether, diethyl ether, n-hexane, acetonitrile, anhydrous sodium sulfate, and methylene chloride were bought from Merck (Darmstadt, Germany). Florisil (PR Grade, 60–100 mesh) was purchased from Silica (Silica Co., USA). All solvents were of pesticide residue grade and subjected to a solvent purity test for residue analysis suitability. Florisil was activated at 130°C for 24 h and cooled to room temperature.
5
Pesticide Extraction and Quantification
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Nickel chloride (99%), zinc chloride (99%), sodium hydroxide (< 90%), and potassium dichromate were purchased from LOBA Chemie, India. Malathion and γ-HCH was purchased from Sigma Aldrich, Germany. From the local University campus, Azadirachta indica leaves were collected. The whole experimental process was performed with deionized water (aqua MAX-Barie 360 series; resistivity 18.2MΩcm -1 ). A stock solution of MLT and γ-HCH having a concentration of 1000 mg L -1 was also prepared.
6
Evaluation of Surfactants and Vegetable Oils for Soil Remediation
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Three non-ionic surfactants, Triton X-100, Tween 80, and Brij L23, were tested in the present study, based on their prevailing use in soil washing (Zheng and Wong, 2010; Zheng et al., 2011) . Surfactants were purchased from Sigma-Aldrich Co. The formula and properties of the surfactants are listed in Supplementary Table . Three vegetable oils were tested to check their suitability as the oil phase, namely soybean oil, the most commonly produced vegetable oil worldwide; linseed oil, the most widely used vegetable oil in industry, and sunflower frying oil, an abundant gastronomic waste with a difficult final disposal. 1-pentanol (purity > 98%, Sigma-Aldrich Co.) was used as cosurfactant.
Lindane [γ-HCH (99% pure)] was purchased from Sigma-Aldrich Co. A stock solution of γ-HCH dissolved in acetone (50 mg mL -1 ) was prepared and employed in all the assays.
Solvents were of pesticide grade, and double distilled water was used for all tests. All other chemicals used throughout this study were of analytical grade and were purchased from standard manufacturers.
Lindane [γ-HCH (99% pure)] was purchased from Sigma-Aldrich Co. A stock solution of γ-HCH dissolved in acetone (50 mg mL -1 ) was prepared and employed in all the assays.
Solvents were of pesticide grade, and double distilled water was used for all tests. All other chemicals used throughout this study were of analytical grade and were purchased from standard manufacturers.
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