BIOWORLD

BIOREMEDIATION HYDROCARBON TREATMENT PRODUCTS

TECHNICAL PRODUCT BULLETIN #B-59
USEPA, OEM REGULATIONS IMPLEMENTATION DIVISION
ORIGINAL LISTING DATE: NOVEMBER 24, 2008
"BioWorld Bioremediation Treatment Products (BioWorld BHTP)"

I. NAME, BRAND, OR TRADEMARK
BioWorld Bioremediation Treatment Products (BioWorld BHTP) Type of Product: Bioremediation Agent (Microbiological Additive)

II. NAME, ADDRESS, AND TELEPHONE NUMBER OF MANUFACTURER/CONTACT
BioWorld USA, Inc. International Headquarters 6734B W Pershing Avenue Visalia, CA 93291 Phone: (559) 651-2042 Fax: (559) 651-9041 E-mail: support@bioworldusa.com Website: http://bioworldusa.com/ (Ms. Diane R. Barnes)

III. NAME, ADDRESS, AND TELEPHONE NUMBER OF PRIMARY DISTRIBUTORS
BioWorld USA, Inc. International Headquarters 6734B W Pershing Avenue Visalia, CA 93291 Phone: (559) 651-2042 Fax: (559) 651-9041 E-mail: support@bioworldusa.com Website: http://bioworldusa.com/ (Ms. Diane R. Barnes)

IV. SPECIAL HANDLING AND WORKER PRECAUTIONS FOR STORAGE AND FIELD APPLICATION
1. Flammability: Non-hazardous. 2. Ventilation: Use adequate ventilation. Treat dry product as a hygroscopic powder and avoid ventilation. 3. Skin and eye contact; protective clothing; treatment in case of contact: Avoid contact with eyes and do not take internally. Upon contact with eyes, flush with plenty of fresh water. Avoid breathing dust. Normal protective equipment for handling of powder, such as a dust mask and eye shield are recommended to be used. Wash hands and body contacted areas with soap and water. 4.a. Maximum storage temperature: 110ºF 4.b. Minimum storage temperature: 34ºF – containers may break if frozen 4.c. Optimum storage temperature range: 34ºF – 90ºF 4.d. Temperatures of phase separations and chemical changes: <32ºF and > 140ºF

IV. SPECIAL HANDLING AND WORKER PRECAUTIONS FOR STORAGE AND FIELD APPLICATION

1. Flammability: Non-hazardous.
2. Ventilation: Use adequate ventilation. Treat dry product as a hygroscopic powder and avoid ventilation.
3. Skin and eye contact; protective clothing; treatment in case of contact: Avoid contact with eyes and do not take internally. Upon contact with eyes, flush with plenty of fresh water. Avoid breathing dust. Normal protective equipment for handling of powder, such as a dust mask and eye shield are recommended to be used. Wash hands and body contacted areas with soap and water.
4.a. Maximum storage temperature: 110ºF
4.b. Minimum storage temperature: 34ºF – containers may break if frozen
4.c. Optimum storage temperature range: 34ºF – 90ºF
4.d. Temperatures of phase separations and chemical changes: <32ºF and > 140ºF

V. SHELF LIFE
The recommended shelf life exceeds 3 years when stored properly in cool, dry place out of direct sun light.

VI. RECOMMENDED APPLICATION PROCEDURE
1. Application Method: BioWorld BHTP is a two part product 1) Bioremediation Enhancer liquid and 2) Hydrocarbon Digesting Microbes in a dry form. Prior to use the microbes are rehydrated in 100ºF water for 10 to 300 minutes. Example for small volume applications – best results are obtained by applying Bioremediation Enhancer first followed by the rehydrated Hydrocarbon Microbe solution in the same location. Example for large volume applications – the two parts of BioWorld BHTP can be mixed with additional water and applied together to spray over the perimeter and over the top of the spill. Example for soil and groundwater applications – evenly apply to soils and mix with till or discing equipment. Oxygen additions are periodically used in certain situations and can be applied with mechanical generation and/or by using oxygen release compound products. 2.Concentration/Application Rate: BioWorld BHTP is applied at an approximate rate of 40 gallons Bioremediation Enhancer liquid and 4 pounds Hydrocarbon Microbes per surface acre treated. Increased quantities and multiple applications may be required to meet shorter cleanup time constraints and desired results. 3.Conditions for Use: BioWorld BHTP can be used in freshwater conditions, wetlands, rock or sand shorelines, contaminated soil areas, salt marshes and salt water conditions (up to 60 ppt salinity).

VI. RECOMMENDED APPLICATION PROCEDURE

1. Application Method:
BioWorld BHTP is a two part product 1) Bioremediation Enhancer liquid and 2) Hydrocarbon Digesting Microbes in a dry form. Prior to use the microbes are rehydrated in 100ºF water for 10 to 300 minutes.
Example for small volume applications – best results are obtained by applying Bioremediation Enhancer first followed by the rehydrated Hydrocarbon Microbe solution in the same location.
Example for large volume applications – the two parts of BioWorld BHTP can be mixed with additional water and applied together to spray over the perimeter and over the top of the spill.
Example for soil and groundwater applications – evenly apply to soils and mix with till or discing equipment. Oxygen additions are periodically used in certain situations and can be applied with mechanical generation and/or by using oxygen release compound products.
2.Concentration/Application Rate:
BioWorld BHTP is applied at an approximate rate of 40 gallons Bioremediation Enhancer liquid and 4 pounds Hydrocarbon Microbes per surface acre treated. Increased quantities and multiple applications may be required to meet shorter cleanup time constraints and desired results.
3.Conditions for Use:
BioWorld BHTP can be used in freshwater conditions, wetlands, rock or sand shorelines, contaminated soil areas, salt marshes and salt water conditions (up to 60 ppt salinity).

VII. TOXICITY AND EFFECTIVENESS

a. Toxicity:

Material Tested	Species	LC50 (ppm)
BioWorld BHTP	Menidia beryllina Mysidopsis bahia	8,848.70 96-hr 7,348.70 48-hr
No. 2 Fuel Oil	Menidia beryllina Mysidopsis bahia	4.50 96-hr 3.21 48-hr
BioWorld BHTP & No. 2 Fuel Oil (1:10)	Menidia beryllina Mysidopsis bahia	13.90 96-hr 7.10 48-hr
Reference Toxicant (SLS)	Menidia beryllina Mysidopsis bahia	15.00 96-hr 15.90 48-hr

b. Effectiveness:
Bioremediation Agent Effectiveness Test (40 CFR 300.900), Federal Register September 15, 1994:

Summary Data Table:

DAYS	PRODUCT 3 REPS/PROD	TOTAL MEAN ALKANES (ppm)	RED% 28 DAYS	TOTAL MEAN AROMATICS (ppm)	RED% 28 DAYS
0	CONTROL NUTRIENT BioWorld BHTP	61,234 64,803 63,192	0 0 0	9,395 8,873 9,220	0 0 0
7	CONTROL NUTRIENT BioWorld BHTP	57,907 49,747 41,480	5 14 28	7,290 5,956 4,405	22 18 40
28	CONTROL NUTRIENT BioWorld BHTP	49,364 21,490 1,948	19 67 97	6,499 5,259 1,078	31 41 88

Results of Gravimetric Analysis:
Percentage (%) Decrease in Weight of Oil on Day 28
Control: 21%
Nutrient: 64%
Product: 96%

VIII. MICROBIOLOGICAL ANALYSIS
1. Listing of all microorganisms by species and percentages in the composition: CONFIDENTIAL 2. Optimum pH, temperature, and salinity ranges for use of the additive: pH: 6-8 Temperature: 80ºF-100ºF Salinity: 0-1 for use of the additive 3. Minimum and maximum pH, temperature, and salinity levels below or above which the effectiveness of the additive is reduced to half its optimum capacity: pH: 3-10 Temperature: 33ºF-120ºF Salinity: >5 Use under any conditions where indigenous populations of oil-degrading microbes are/are not present 4. Special nutrient requirements: Some sites may benefit with various nutrients added depending on project variables 5. Test results regarding the determination of the presence of the following: Salmonella: Negative Fecal coliform: <2 Shigella: Negative Staphylococcus Coagulase positive: <10 Beta hemolytic Streptococci: <10

VIII. MICROBIOLOGICAL ANALYSIS

1. Listing of all microorganisms by species and percentages in the composition: CONFIDENTIAL
2. Optimum pH, temperature, and salinity ranges for use of the additive:
pH: 6-8
Temperature: 80ºF-100ºF
Salinity: 0-1 for use of the additive
3. Minimum and maximum pH, temperature, and salinity levels below or above which the effectiveness of the additive is reduced to half its optimum capacity:
pH: 3-10
Temperature: 33ºF-120ºF
Salinity: >5
Use under any conditions where indigenous populations of oil-degrading microbes are/are not present
4. Special nutrient requirements: Some sites may benefit with various nutrients added depending on project variables
5. Test results regarding the determination of the presence of the following:
Salmonella: Negative
Fecal coliform: <2
Shigella: Negative
Staphylococcus Coagulase positive: <10
Beta hemolytic Streptococci: <10

IX. PHYSICAL PROPERTIES
NA

X. ANALYSIS FOR HEAVY METALS, CYANIDE, AND CHLORINATED HYDROCARBONS
NA

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BIOREMEDIATION HYDROCARBON TREATMENT PRODUCTS

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