®
Trivol
Bioremediation
Ameret, LLC
Environmental review
4/13/2015
1
Introduction
Ameret, LLC has been working with the top microbiologists with over
two decades of experience to develop a series of bacterial cultures to
enhance the biological breakdown process of hydrocarbons for the Oil
and Gas Industry
Ameret, LLC utilizes a third party fermentation company that
specializing in the development and production of microbial products for
improved environmental processes.
Trivol® Bioremediation technologies differ significantly from
competitive products in fermentation, culture type, and product
capabilities, they are among the most sought after product for agriculture,
water restoration, wastewater and environmental processes.
.
4/13/2015
2
Ameret, LLC






Latest technologies and equipment
Highly scientific personal, all levels
Significant production capacity (demand)
Ready to ship inventory (in stock supplies)
Quality Assurance testing of in coming materials
to finished product (quality)
Technical support
4/13/2015
3
Trivol® Bioremediation Products






www.oilspillremediationproducts.com
Culture Isolation, Selection & Enhancement
Formulation Development
Laboratory and field confirmation of product
performance
Trivol® Bioremediation products are non-toxic, safe
for use around animals, pets, wildlife and humans.
Truly the most environmentally friendly products
available.
4/13/2015
4
Trivol® Bioremediation Products




Completely different technology
Different group of microorganisms
Different fermentation process
Fermentation - Six day multi stage, final five day
facultative process
Key Factor…
 The only facultative culture consortium on the
market !
4/13/2015
5
Safety & Performance
Toxicity data – clearly your best choice
No Toxicity
Straight fermentation
Enzyme free
No chemicals
Novel Biological Products
Formulations With Unmatched Oxidation Performance Capabilities
4/13/2015

Trivol® HDM (liquid)

Trivol® Sludge Away (liquid)
7
Proven Technology for Bioremediation

Trivol® Bioremediation technology are two-fold

1) Trivol® HDM - a select consortium of Purple Sulfur microorganisms
designed for anaerobic environments, degrades a wide range of
compounds, including those that contain hydrocarbons

2) Trivol® Sludge Away – a natural non-chemical humas-based-soilscience-extract designed to speed degradation of dense
hydrocarbons and improve ecosystems building value

The combination has proven effective in oil and gas bioremediation in
aerobic and anaerobic conditions

Ameret, LLC offers products and technical support for a wide range
of oil and gas spill problems.
Trivol® Bioremediation product’s multi stage fermentation , and five day anaerobic
process enhances anaerobic functions result in cultures capable of enhancement and
controlling gaseous reaction
Background - Fermentation in anaerobic environments is an energy-releasing process
that takes place in the absence of oxygen, resulting in the breakdown of carbohydrates
and complex organic substances into simpler matter. Only certain organisms perform
fermentation to obtain energy to carry on these processes.
Few to no competitive products obtain energy through fermentation, these products are
designed to gain energy through aerobic respiration in the presence of free oxygen.
These spore based products are produced in single stage aerobic processes.
Trivol® Bioremediation capabilities are derived by type of microorganism carrying on
the process, and their development process to achieve necessary anaerobic reactions.
Oil pits are strict anaerobic environments where fermentation by microorganisms
produce substances called enzymes which speed up chemical reactions to breakdown
matter. Trivol® Bioremediation is designed to enhances these functions
4/13/2015
9
®
Trivol

Trivol® Bioremediation is designed for Oil and Gas industry and differs
from completive formulations. This culture consortium is effective in all
ecosystems. The consortium consists of spore and vegetative bacteria.
What is the difference?



Bioremediation
Activity level – spore-forming organisms can choose to ‘shut down’ and go to
spore if they do not like their environment; vegetative bacteria cannot go to
spore so they must manipulate their environment to meet their needs
Functions – vegetative bacteria are much more dynamic in their capabilities
over spore-forming organisms
Trivol® Bioremediation provides a reduction in any hydrocarbon on land
or water and provide related plant benefits by balancing the ecosystems
biology.
4/13/2015
10
Addressing Environmental Concerns

The environmental effects of oil spills are a
concern, particularly with respect to surface water
and groundwater quality and to air quality as
affected by odors and gaseous emissions from
large-scale operations

Trivol® Bioremediation addresses most of these
issues
4/13/2015
11
Nutrient Retention (Nitrogen)

Oil pits and contaminated soil may remove a
significant amount of Nitrogen, but Phosphorus
and Potassium are not likely to be significantly
affected by treatment.

Trivol® Bioremediation technology builds biomass
that stores nitrogen and significantly reduces the
potential 80% loss of nitrogen within ecosystems
4/13/2015
12
Building Soil Biology and Nitrogen
Values



The key to estimating N availability after application
rests on mineralization decay rate expected from the
breakdown of organic matter.
For crude oil the estimated first-year contribution
from organic N can vary from 25 to 50% of the total
organic N. This is a function of the breakdown rate of
oil in the soil.
Trivol® Bioremediation accelerates these reactions,
and stores released nutrients in ecosystem
4/13/2015
13
Biological Control of Runoff

In many areas there are major concerns associated
with managing oil spills related to runoff control

Trivol® Bioremediation bio technology controls
runoff, reduces water born pathogens, reduces
nitrate in water via denitrification, and reduces
organic matter in field trenches

Trivol® Bioremediation releases nutrient energy
contained in hydrocarbons for plant up-take
4/13/2015
14
Microbial Functions

Biological enzymatic reactions are involved in the
liquefaction, breakdown and stabilization of any
hydrocarbon

Trivol® Bioremediation technology considers the
anaerobic and facultative environmental conditions
in ecosystems
4/13/2015
15
Microbial Oil Pit Management

To understand bio-augmentation a review of the role of microorganisms
and how bacteria function is necessary.

Microorganisms are responsible for the enzymatic breakdown,
conversion and removal of organic matter from the environment, this
biological conversion of oil is all the way to the basic elements of carbon
dioxide, water and recycled nutrients (Bio-Mass)

Microbes play a key role in breaking down oil and other organic matter in
ecosystems

BIOLOGICAL TREATMENT ,

This is a review of microbial enzymatic process and how these select
microorganism can enhance the soil or water via improved enzyme
processes.
4/13/2015
16
Microbes in Action
How Microorganisms Function in field use reducing and converting
organic matter to energy for plant up-take
4/13/2015
MICROBIOLOGY and waste
management
17
Bio Enzymatic Reactions
A biological reduction process to convert matter to energy for pants





Extracellular enzyme – are secreted by the cell to
breakdown larger materials prior to entering the cell as
food
Membrane bound enzymes –breakdown dissolved
and particulate compounds attached to the cell wall
Intracellular enzymes – act on dissolved and
compounds that cross the cell membrane
Bio surfactants – are also secreted to emulsify
compounds such as grease, fats, fatty acids and even
petroleum hydrocarbons
TIME PROCESS!
Understanding
Bio-Enzymatic Reactions That Degrade Hydrocarbons
Extracellular Enzyme
C02


cell bound enzyme
H20
Intracellular enzyme
4/13/2015
19
Bio-Enzymatic Reactions
A review of how microbial cells grow and divide during organic conversion




Oxidation organic matter- CxHyOz – Enzyme ->
CO2 + H2O
Cell synthesis (growth) CxHyOz + NH3 + P04 +
02 – Enzyme -> Cells + CO2 + H20
Oxidation cell material (cells) + 02 – Enzyme ->
CO2 + H20 = NH3
(Weston and Eckenfelder, 1995)
4/13/2015
20
BINARY FUSION - Cell Growth and Division
Building Microbial Bio-Mass and Ecosystem Value
4/13/2015
MICROBIOLOGY and waste
management
21
Bio-Augmentation

Now you know oil pits rely on enzyme reaction and
microbiology to keep degrading hydrocarbons.

Biological enzyme activity is a key factor in remediation.

Trivol® Bioremediation offers significant benefits in
hydrocarbon remediation from “top-to-bottom” via consistent
enzymatic breakdown of oil.

Bio-augmentation with these select cultures significantly
improve hydrocarbon remediation.
4/13/2015
22
Growth
Curve
Building Bio Mass for Improved ecosystem stability and value on application,
look at lag to log growth indicating a significant increase in Bio-Mass during oil
storage. The declining phase takes place on land application during conversion
of matter to plant values
23
Growth Conditions Plus






pH – 6 to 8 (7.5 opt)
Macro Micronutrients C. N. P = 00 - 5 – 1
Ammonia nitrogen (NH3 (N) -1/mg/r) ,
Ortho-phosphate (PO4) – 1 mg/r)
Temperature (F) 55-110 (opt 70/85)
Not important just a review of oxidation reaction
on field application of Trivol® Bioremediation
products
4/13/2015
24
Soil Webb
Crops and Microbe Association




Plants produce carbohydrates to generate root
microbial contact
Soil bound organic matter released by bio
enzymatic functions
Microbes bind to soil particles and root zones
Trivol® Bioremediation technologies build biomass to enhance the ecosystem as well as retain
nutrients
4/13/2015
25
Microorganisms are the foundation in the soil web
See the first two trophic levels
26
Root systems are the foundation of plants
No Treatment vs. Treated
Great Root and Shoot Growth
Treated 64oz / acre @ planting
4/13/2015
30
Increased yield = 38 to 53
4/13/2015
31
Biological Nutrient Retention
4/13/2015
32
Microorganisms change their polymer production
4/13/2015
34
Create binding detainment films
for improved nutrient storage
35
Field Crop Benefits







4/13/2015
Increases soil & plant biology
Stimulate root zones and rhizosphere
Bind nutrients and hold moisture
Controls nutrient run off
Fixes Nitrogen from the atmosphere, retains
soil values
Increase plant health
Increase crop yield
36
NITRIFICATION !



Due to a poor understanding of soil biology and
plant relationships with these reactions, Nitrification
inhibitors have been considered to arrest the rate
of mineralization; however, they have not been fully
evaluated under field conditions and the results
have been variable.
Trivol® Bioremediation cultures drive essential
nitrification and provide added sources of energy
for plants, i.e.,  ammonia to  nitrite- to 
nitrate with plant utilization of these elements.
And don’t forget other biological contributions.
4/13/2015
37
THANK YOU!
4/13/2015
38