THE USE OF BENEFICIAL MICROORGANISMS IN CLEANING FORMULATIONS

Microorganisms overview 2 Agenda Microorganisms overview Microorganisms and chemistry What can you do with microorganisms? Focus on drain application Regulatory

MICROORGANISMS OVERVIEW

MICROORGANISMS ARE LIVING CREATURES FOUND EVERYWHERE ON EARTH… 4 MICROORGANISMS ARE LIVING CREATURES FOUND EVERYWHERE ON EARTH… Only 10% of the cells in the human body are human! Earth 5 nonillion (5×1030) Ocean 10 million in a ml. of water Soil 40 million in a gram of soil Human Body 40 million in a ml. of saliva 100 trillion in the gut flora 10% of total dry weight

THE VAST MAJORITY OF MICROORGANISMS ARE EXTREMELY SAFE 5 THE VAST MAJORITY OF MICROORGANISMS ARE EXTREMELY SAFE In fact, we not only live with microbes, but our health is dependent on them (gut bacteria, immune system development, etc)

WHAT ARE BACTERIA? Bacteria are organisms that are: 0.5 – 3µm Living 6 WHAT ARE BACTERIA? 0.5 – 3µm Bacteria are organisms that are: Living Single-celled Microscopic

BACTERIAL NUTRITION/ ENVIRONMENTAL CONDITIONS 7 BACTERIAL NUTRITION/ ENVIRONMENTAL CONDITIONS Carbon, Nitrogen, Phosphorus and some trace of minerals to grow Moisture, proper temperature, appropriate pH and something to “breathe” (an electron acceptor like O2, NO3, etc.) Production of enzymes (biocatalysts) that break down compounds like fat, starch and protein to yield these essential nutrients

COMPARISON OF BACTERIA AND ENZYMES 8 COMPARISON OF BACTERIA AND ENZYMES Raw enzymes are... fast starting biochemicals specific for a certain reaction proteins that can be degraded by other enzymes Bacteria... take a bit longer awaken and to show activity produce enzymes on demand, in response to compounds present (in situ enzyme factories) can produce suite of enzymes to completely degrade a compound can use the components and energy from the enzymatic action to produce more bacteria

BACTERIA PRODUCE ENZYMES 9 BACTERIA PRODUCE ENZYMES IN PART, ENZYMES ARE USED TO FEED THE BACTERIA Extracellular enzymes Large molecules, particulate or colloidal material Small, soluble compounds Intracellular enzymes CO2 & H2O Only ~3% of enzymes used by microorganisms are secreted; the rest are accessible for useful applications only by using the whole cell

MICROORGANISMS AND CHEMISTRY

CHEMISTRY/ENZYMES/MICROORGANISMS 11 CHEMISTRY/ENZYMES/MICROORGANISMS SURFACTANT/ SOLVENT Cleaning by displacement and solubilization of organic soils Increase the efficiency of enzymes and microorganisms by contact improvement ENZYMES Cut organic soils /catalyze chemical reaction Facilitate the rolling- up and cleaning (surfactant, mechanical action…) MICROORGANISMS Enzyme factory Metabolize/ degrade organic substrates Minimize impact on the environment + H2O + CO2 Same organic content Same organic content Organic content decreased

FORMULATE WITH MICROORGANISMS 12 FORMULATE WITH MICROORGANISMS pH: 5.0 to 9.0 for optimal stability in liquids Easy to formulate but need basic expertise (living technology)

SO, WHAT CAN YOU DO WITH MICROORGANISMS?

MICROORGANISMS BENEFITS 14 MICROORGANISMS BENEFITS Short term cleaning: chemistry and enzymes Mid-term and long term degradation: microorganisms Microorganisms Continuous organic degradation (fat, starch, protein, VFA) Chemistry Enzymes Fight stains Odor control < 1 min 1 hour Time

CURRENT CLEANING SOLUTIONS APPLICATIONS 15 CURRENT CLEANING SOLUTIONS APPLICATIONS Carpet & fabric Improved cleaning Decrease resoiling Drain Avoid blockages Odor control Grease Interceptor Odor control Prevent grease release in the sewer Hard surface Deep cleaning Odor control Odor control Drain Hard surface Grease interceptor Septic & waste Septic & waste Odor control Reduce blockages Enhance the degradation

FOCUS ON DRAIN APPLICATION

KITCHEN AND BATHROOM SOIL CLEANING 17 KITCHEN AND BATHROOM SOIL CLEANING Effective soil cleaning Before treatment with Green Worx's technology Water rinse only Treated with Green Worx's technology Green Worx’s technology effectively removes both bathroom soil (left) and kitchen soil (right) applied to ceramic tiles.

18 KITCHEN SOIL CLEANING Green Worx CSs’ technology effectively removes kitchen soil (right) while the control without treatment (left) shows no change

KITCHEN SOIL CLEANING Free flowing drain line 19 KITCHEN SOIL CLEANING Untreated (kitchen soil only) Treated with GW microorganisms The tube treated with Green Worx’ microorganisms (right) shows almost complete soil removal, and the time for water to flow through the tube improved by 25 seconds. Free flowing drain line

MICROBIAL ACTIVITY ON KITCHEN SOIL 20 MICROBIAL ACTIVITY ON KITCHEN SOIL Respirometric activity The long-lasting, continuous degradation (15+ days) of kitchen soil and grease is demonstrated by the respirometric activity of Green Worx’ beneficial microorganisms.

21 PROTEIN DEGRADATION After 7 days of treatment with Green Worx s’ beneficial microorganisms, a black protein cube is degraded and black dye is released (right) whereas the control (left) shows no protein degradation.

22 STARCH DEGRADATION Complex starch degradation before (left) and 72 hours after (right) addition of Green Worxs’ beneficial microorganisms.

ODOR CONTROL - MICROBIAL GROWTH ON 23 ODOR CONTROL - MICROBIAL GROWTH ON ODOROUS MOLECULES AND BUILDUP Respirometry test on VFA Green Worxs’ beneficial microorganisms can degrade both the long- and the short-chain fatty acids commonly found in kitchen drain soil and responsible for malodors and buildup.

REGULATORY

REGULATORY AND SAFETY INFORMATION 25 REGULATORY AND SAFETY INFORMATION Tight quality control and stringent tests are incorporated into the production process to ensure that Green Worx CS products are manufactured to the highest quality standards All microorganisms are isolates of naturally occurring organisms and are not genetically engineered or modified All microorganisms have been identified by 16s rDNA sequencing and confirmed to belong to Risk Group 1 as defined by the Directive 2000/54/EC. Safety assessment performed on each Green Worx CS product prior to market Reach compliance of all raw materials is checked .

26 CONCLUSION Beneficial bacteria can degrade overtime, making preventive treatment really efficient Spore-forming beneficial bacteria have long shelf life Bacteria are sustainable and less toxic than traditional chemical control agents Existing of a large collection of bacteria-based products used for degradation of grease, oil, odors in kitchen drains and carpets, organic soiling/stains, etc. Green Worx CSs’ beneficial microorganisms could offer safe, sustainable, efficient solution to any application where control of undesirable compounds is needed

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