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Kocide 3000

Manufactured by DuPont
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Sourced in United States

Kocide® 3000 is a copper-based fungicide and bactericide developed by DuPont. It is a water-dispersible granule formulation that contains 46.1% copper as the active ingredient.

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Lab products found in correlation

Cupric acetate monohydrate, by Sinopharm (1 mentions) Glycine, by Sinopharm (1 mentions) Live dead baclight bacterial viability kit, by Thermo Fisher Scientific (1 mentions) Eclipse ti, by Nikon (1 mentions) Nis elements imaging software, by Nikon (1 mentions) Nitric acid, by Thermo Fisher Scientific (1 mentions) Ethanol, by Thermo Fisher Scientific (1 mentions) Copper 2 sulfate pentahydrate, by Thermo Fisher Scientific (1 mentions) Tetraethylorthosilicate teos, by Gelest (1 mentions) Icap 6300, by Thermo Fisher Scientific (1 mentions) D8 advance, by Bruker (1 mentions) Permethrin, by Merck Group (1 mentions) Cyclohexane, by Merck Group (1 mentions) N butyl acetate, by Merck Group (1 mentions) Copper 2 sulfate pentahydrate cuso4 5h2o, by Merck Group (1 mentions) Hexadecyltrimethylammonium bromide, by Merck Group (1 mentions) Acetonitrile, by Merck Group (1 mentions) Methanol, by Merck Group (1 mentions) Ethanol, by Merck Group (1 mentions) Calcium chloride, by Merck Group (1 mentions)

7 protocols using kocide 3000

1

Copper Hydroxide Synthesis and Xanthomonas Strain

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Cupric acetate monohydrate and glycine (Gly) were supplied by Sinopharm Chemical Reagent Beijing Co., Ltd. (Beijing, China). Copper(ii) hydroxide (99%, Chengxin Industry CO. LLC, Shanghai, China), Kocide 3000 (46.1 wt%, copper hydroxide) was supplied by DuPont (Wilmington, DE, USA). Xanthomonas campestris pv. campestris (Xcc) strain was obtained from the College of Plant Science, Tarim University (Alaer, China).
Dong H., Xiong R., Liang Y., Tang G., Yang J., Tang J., Niu J., Gao Y., Zhou Z, & Cao Y. (2020). Development of glycine-copper(ii) hydroxide nanoparticles with improved biosafety for sustainable plant disease management. RSC Advances, 10(36), 21222-21227.
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2

Viability Assay for Xanthomonas perforans

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X. perforans strain GEV485 was used for the viability assay. Bacterial cells were incubated in nutrient broth at 28 °C on a shaker at 300 rpm for 16 h and harvested in the log phase. Bacterial cells were pelleted by centrifugation (5000 rpm for 10 min) and resuspended in 0.01 M MgSO4·7H2O, and the suspensions were adjusted to A600 = 0.3 (~5 × 108 CFU/mL). Then, 500 μL of the bacterial suspension were transferred to 4.5 mL of Kocide® 3000 (DuPont, Wilmington, DE, USA) at 1000 µg/mL. Sterilized tap water served as the control. The tubes were incubated at 28 °C on a shaker at 300 rpm for 4 h. After washing with 1 mL of 0.85% NaCl twice, 1 mL samples from each tube were stained using the LIVE/DEAD BacLight Bacterial Viability kit (L7007, Molecular Probes (Eugene, OR, USA), Invitrogen (Waltham, MA, USA)). The stain was a mixture of 1.5 mL Component A and 1.5 mL Component B. Following addition, the sample was incubated in darkness for 15 min at room temperature. Micrographs were taken with a Nikon Eclipse Ti inverted microscope (Nikon, Melville, NY, USA) at ×40 fluorescent optics using NIS-Elements imaging software (Ver. 3.0; Nikon). The dead cell/all cell ratio was calculated by ImageJ [34 (link)].
Liao Y.Y., Pereira J., Huang Z., Fan Q., Santra S., White J.C., De La Torre-Roche R., Da Silva S., Vallad G.E., Freeman J.H., Jones J.B, & Paret M.L. (2023). Potential of Novel Magnesium Nanomaterials to Manage Bacterial Spot Disease of Tomato in Greenhouse and Field Conditions. Plants, 12(9), 1832.
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3

Copper-Based Nanocluster Pesticide Protocol

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PEG@Cu NCs, a copper nanocluster pesticide, was provided by Professor Kun Qian from Southwest University of China (Chongqing, China). Kocide® 3000 (46% copper hydroxide water-dispersible granule) is a commercial pesticide produced by DuPont Company (Wilmington, DE, USA), and 98% copper acetate anhydrous was purchased from Shanghai Macklin Biochemical Co., Ltd (Shanghai, China). Reconstituted water of pH 7.5 ± 0.5 was prepared according to the guideline of ISO-7346-2 with minor adjustments, which mainly contained 294 mg/L CaCl2 · 2H2O, 106.5 mg/L NaHCO3, 60 mg/L MgSO4 and 6 mg/L KCl [20 ].
Gao F., Yuan Z., Zhang L., Peng Y., Qian K, & Zheng M. (2023). Toxic Effects of Copper Fungicides on the Development and Behavior of Zebrafish in Early-Life Stages. Nanomaterials, 13(19), 2629.
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4

Copper-Based Nanocluster Pesticide Protocol

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PEG@Cu NCs, a copper nanocluster pesticide, was provided by Professor Kun Qian from Southwest University of China (Chongqing, China). Kocide® 3000 (46% copper hydroxide water-dispersible granule) is a commercial pesticide produced by DuPont Company (Wilmington, DE, USA), and 98% copper acetate anhydrous was purchased from Shanghai Macklin Biochemical Co., Ltd (Shanghai, China). Reconstituted water of pH 7.5 ± 0.5 was prepared according to the guideline of ISO-7346-2 with minor adjustments, which mainly contained 294 mg/L CaCl2 · 2H2O, 106.5 mg/L NaHCO3, 60 mg/L MgSO4 and 6 mg/L KCl [20 ].
Gao F., Yuan Z., Zhang L., Peng Y., Qian K, & Zheng M. (2023). Toxic Effects of Copper Fungicides on the Development and Behavior of Zebrafish in Early-Life Stages. Nanomaterials, 13(19), 2629.
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5

Copper-tolerant Bacterial Strains Isolation

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All reagents were obtained from commercial vendors and used without further purification. These include: tetraethyl orthosilicate (TEOS, 98%, Gelest Inc., PA, USA), didecyldimethylammonium chloride (DDAC, 50% solution in 2-propanol/water 2 : 3 from EMD Millipore, MA, USA), ethanol (190 proof, Pharmco, CT, USA), ethanol (200 proof, Acros Organics, NJ, USA) and copper(ii) sulfate pentahydrate (99+%) (Acros Organics, NJ, USA), nitric acid (68 to 70% (w/w), Fisher Scientific, PA, USA), sodium hydroxide solid beads (Fisher Scientific, PA, USA), nutrient broth (NB) and agar (Fluka, St. Louis, MO, USA). Xanthomonas alfalfae subsp. citrumelonis (ATCC 49120), Pseudomonas syringae pv. syringae (ATCC 19310), and Clavibacter michiganensis subsp. michiganensis (ATCC 10202) cultures from ATCC (U.S. Department of Agriculture (USDA) permits P526P-12-04060 and P526P-15-01601) and X. perforans strains GEV485 (Cu-tolerant) and 91-118 (Cu-sensitive) isolated from tomato in Florida provided by Paret and Jones (North Florida Research and Education Center, University of Florida, USA) were used. Kocide 3000® (DuPont™) is a Cu-based fungicide/bactericide retaining 46.1% copper hydroxide (CAS No. 20427-59-2) available for commercial use for growers.
Ozcan A., Young M., Lee B., Liao Y.Y., Da Silva S., Godden D., Colee J., Huang Z., Mendis H.C., Campos M.G., Jones J.B., Freeman J.H., Paret M.L., Tetard L, & Santra S. (2021). Copper-fixed quat: a hybrid nanoparticle for application as a locally systemic pesticide (LSP) to manage bacterial spot disease of tomato. Nanoscale Advances, 3(5), 1473-1483.
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6

Characterization of Copper Nanomaterials

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The Cu particles used in this study include nanoparticulate CuO (denoted nCuO), micron-sized Cu and CuO (denoted bulk Cu and CuO respectively; all obtained from Sigma Aldrich), nanoparticulate Cu (US Research Nanomaterials, denoted nCu), Kocide 3000 (Dupont, Wilmington, DE), and CuPRO 2005 (SePRO, Carmel, IN). Reagent grade CuCl2 salt was obtained from Sigma Aldrich. The size and surface charge of particles at pH 7 (0.5 mM phosphate buffer) were determined by measuring hydrodynamic diameter (HDD) and zeta (ζ) potential using a Zetasizer Nano-ZS90 (Malvern, UK). A previous study33 showed that phosphate buffer, at the concentration used, had only minimal effects on zeta potential measurements. Primary particle size and morphology were determined via scanning electron microscopy (FEI XL40 Sirion) equipped with an Oxford INCA energy-dispersive X-ray spectroscopy (EDS) probe. Copper content (wt. %) of each particle was determined via ICP-AES (iCAP 6300, Thermo Scientific).34 (link), 35 The main copper phase and crystal structure of particles were determined via X-ray diffraction (XRD, Bruker D8 Advance).
Cu NPs/compounds’ suspensions/solutions were prepared at 0, 5, 10, and 20 mg/L in modified Hoagland’s nutrient solution36 (link) and homogenized by sonication in a water bath (Crest Ultrasonics, Trenton, NJ) at 25 °C for 30 min. There were four replicates per treatment.
Hong J., Rico C., Zhao L., Adeleye A.S., Keller A.A., Peralta-Videa J.R, & Gardea-Torresdey J.L. (2014). Toxic Effects of Copper-based Nanoparticles or Compounds to Lettuce (Lactuca sativa) and Alfalfa (Medicago sativa). Environmental science. Processes & impacts, 17(1), 177-185.
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7

Nanopesticide Preparation and Evaluation

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Calcium chloride (CaCl2, 99.9%), sodium carbonate (Na2CO3, 99.9%), N-butanol (C4H9OH, 99.9%), cyclohexane (C6H12, 99.9%), methanol (CH3OH, 99.9%), ethanol (CH3CH2OH, 99.9%), acetonitrile (CH3CN, 99.9%), validamycin (C20H35NO13, 73.7%), dichloromethane (CH2Cl2, 99.9%), polyethylene glycol (PEG) (H(OCH2CH2)nOH, ≥98), n-butyl acetate (C16H12O2, 99.9%), sec-butyl alcohol (C4H10O, 99%), copper(II) sulfate pentahydrate (CuSO4·5H2O), and calcium carbonate (CaCO3) were purchased from Merck (Darmstadt, Germany). Ethyl methanesulfonate (EMS, CAS No. 62-50-0), hydrogen peroxide (H2O2, CAS No. 7722-84-1), acephate (C4H10NO3PS, ≥98), permethrin (C21H20Cl2O3, ≥90), hexadecyl trimethyl ammonium bromide (C19H42BrN, ≥98), phosphatidylcholine (from soybean lecithin) (C42H80NO8P, 40%), and ammonium glycyrrhizate (C42H62O16.NH3, ≥70) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Copper hydroxide (Cu(OH)2) particles were purchased from DuPont as a commercial biocide (Kocide® 3000). For negative control groups, we used sterile distilled water, ethanol (2%), nanocapsules (2.5 mM), nanocapsules (5 mM PEG-400), and nanocapsules (2.5 mM CaCO3) for the preparation of the CuSO4·5H2O and Cu(OH)2 nanopesticides, permethrin, permethrin nanopesticides, acephate nanopesticides, and validamycin nanopesticides, respectively.
Demir E., Kansız S., Doğan M., Topel Ö., Akkoyunlu G., Kandur M.Y, & Turna Demir F. (2022). Hazard Assessment of the Effects of Acute and Chronic Exposure to Permethrin, Copper Hydroxide, Acephate, and Validamycin Nanopesticides on the Physiology of Drosophila: Novel Insights into the Cellular Internalization and Biological Effects. International Journal of Molecular Sciences, 23(16), 9121.
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