Culture Your Own Purple Non-Sulfur Bacteria! Provides Superior Nutrition Helps Control Nutrients and Organic Waste Full 2.5 Gallon Culture Treats up to 8000 Gallons PNS HomeGro contains key items required to economically cultivate purple nonsulfur bacteria (PNSB) using commonplace chemicals and implements. This unique kit presents a more eco-friendly alternative to relatively small, pre-grown bottled products as it significantly reduces shipping/packaging waste. Finished culture treats approximately 8,000 gallons! Each PNS HomeGro Kit includes an inoculum bottle with: Rhodopseudomonas palustri Rhodospirillum rubrum Rhodobacter spheroides What's Included? Culture Vessel, Cap, and Spigot Sticker Thermometer Growth Medium Powder Inoculum Bottle (select blend of probiotic PNSB cultures) Other Supplies Needed (not included): Kitchen Funnel Kitchen Thermometer Unscented Household Bleach Hydrogen Peroxide (3% or higher) w/ Spray Bottle Isopropyl Alcohol (70% or higher) w/ Spray Bottle 60-65W Incandescent Lamp or Equivalent Heat Pad (if needed for temperature control) DIRECTIONS Prepare Work Area Sanitize Hands and Work Surfaces Establish a clean, uncluttered, draft-free, and dust-free area. Wash hands/gloves thoroughly with antibacterial soap. Avoid loose, long-sleeve clothing (we further recommend wearing face mask and tying back long hair). Sterilize the Culture Vessel Pour 1 capful of household bleach into the vessel (use the vessel cap for measurement, as this will sterilize the inner surfaces of the cap). Add 1 cup of purified water to the vesselecap the vessel and shake vigorously. Uncap and pour out bleach water, shaking out any remaining drops (do not rinse). Without touching the inside of the cap, immediately recap the vessel. Prepare Growth Medium Boil Water and Sterilize Tools Bring 3 gallons of purified water to a boil in a large pot with lid. Sterilize implements (stirring spoon, thermometer, etc.) by spraying them with peroxide and placing them on a sterile surface (e.g., a tray that has been sterilized with peroxide). Mix Growth Medium Slowly add the growth medium powder to the boiling water and stir well. Cover pot and maintain boil for 10 minutes; adjust heat as necessary to avoid boilover. Turn off the heat and allow pot to cool to 90°C (194°F); this temperature is hot enough to discourage biocontaminants, but not hot enough to melt the culture vessel. Transfer to Vessel Using a sterilized funnel, carefully pour the mixture into the culture vessel. For ease and safety, you may use a sterilized pitcher to transfer the liquid to the vessel (from pot to pitcher to vessel); sterilize pitcher with peroxide, then rinse the peroxide away with isopropyl alcohol, before use. On the final pour, stir up any settled growth medium in the pot to ensure that it is transferred to the culture vessel. Without touching the inside of the cap, immediately recap the vessel. Inoculation and Incubation Cool Vessel/Add Inoculum Allow the vessel to cool to room temperature; generally, this requires that the vessel is set aside overnight before starting next step! Do not add inoculum to hot culture medium! Lightly shake inoculum bottle. Remove cap and carefully pour the entire contents of the inoculum bottle into the vessel (do not touch inside of cap). Immediately recap the vessel. Incubate Culture It is critical to incubate at the appropriate temperature; maintain steady temperature of 28°-30°C (82°-86°F); use a heat pad with thermostat, or similar controllable heat source, as required. Apply sticker thermometer to vessel (opposite side of vessel from main heat source); secure with clear tape if necessary. Place the vessel under continuous, moderate light; a 60-65 watt incandescent (or equivalent ‘warm white’ LED) light source (about 1 inch from vessel) is ideal. Monitor Growth Complete incubation takes approximately 30 days (often longer). To accelerate growth, shake the vessel (preferably daily) as reddish sediment accumulates on the bottom. The culture is fully mature when no additional growth occurs for 5+ consecutive days (usually after one month); it is common for much of the living biomass to settle to the bottom at this time. Prepare for Use Sterilize spigot with peroxide, then rinse the peroxide away with isopropyl alcohol. Replace cap with sterilized spigot (avoid touching inside of spigot). Shake the vessel before each use. Spray nozzle of spigot with peroxide, then isopropyl alcohol, before and after each use.
