Note: Descriptions are shown in the official language in which they were submitted.
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PRE-ACCLIMATIZATION AND AMPLIFICATION SYSTEM FOR
MICROORGANISMS
BACKGROUND OF THE INVENTION
With the increasing population concentration in most suburban and urban area doubled with
the more and more restrictive local, national and international en~ onl-.enlal protection laws,
it is becoming m~nrl~tory to collect the waste water sewage from the individual residentials,
commercial and industrial sewage ~m~ent~ and to process it in a central location before it is
relargued back into the ecosystem.
If left untreated or hlappl upliately treated, most types of sewage effluents will exceed the
discharge limits and thelerol~ will not be in compliance with the environmental laws. Most
effluent will also genel~le foul odors ( NOx and SOx ) wich can be, in the best case, abnoxious
for the workers and, in the worst case, life ll~ealn~llg for the workers and people living next
to the processing area when those gaz colllb;l,e with others to form toxic emanations ( Shrapnel
effect ).
The overload of organic wastes such as grease, proteins, starch and any other organic wastes,
have a tendency to accl-mul~te and form a thick crust at the air water interface, that will clog
the collection system at one or many critical points. This issue is generally corrected by
r.~ue..~ and costly mecanical removals ofthe excedentary organic accumulations. This solid
acc Imlll~tion also, often l-~fo..~ an anaerobic pe-ro~-lla..l system into a poor and ineffective
anaerobic foul odors gene.~li.lg system .
At one point, with the increased volume of organic wastes generated by the increase in the
population and/or econo.. ical activity, a previously appropliale sewage treatment plant may
become unable to process the increase of organic load found in the effluent. Therefore
significant capital in~/~l.ne..l will be nece~a.y in order to comply with the environmental laws.
Most of the time this in~e~t...e..l tr~n~l~tes into the buying of larger equipments that are only
providing a better retention time and a better dilution factor in the effluent without really
solving the problem at the source .
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Most countries have put into place more or less severe limitations on the quantities of organic
and inorganic matters which may be introduced back into the municipal sewage collection
system. For hl~lallce~ every business and/or industrial co~..panies gene,~ling large quantities
of organic and inorganic matters are required to install a retention device wich may be called
septic tank, grease trap, aerated lagoons, pumping station, etc. In most cases, those retention
time devices are treated mecanically and/or with a chemical and/or with a biological approach
which indudes the daily addition or multiple injections of selected strains of bacteria in order
to accelerate the speed of waste biodegradation.
As a matter of fact, the continuous addition of selected strains of bacteria may reduce the
organic and /or inorganic load by converting the waste products into lower molecular weight
componds with less or no envi~(,r.l.~elllal impact via the in-situ and ex-situ biodegradation
ability of ba~;le~ias. Because of it's l-~..,.n-lu~.s ellvilolllllelll~l potential, this approach has been
exploited and improved over the recent years. The addition of bacteria called Bio-
Aug~ lion and/or Bio-remediation is now a well known and accepted technic in the field
of soils re...edialion, air remediation and water deco~ ion. The application of the Bio-
Augmentation technic which requires daily and/or frequent addition of a large number of
selected strains of bacteria may become, in a long term, costly and hence not as profitable for
the users.
The Applicant has noticed that this Bio-~ ion approach is not effective because of the
small popuiation of microor~i.ems in front of a large amount of waste products to degrade.
This is due to the con~t~..l wash out of microolg~-;.e...~ by the flow of the waste water
collector. Hence, the conv~llliollllal application of Bio-Augmentation and/or Bio-remediation
is not an efficient and/or econo--Lcal way to elimin~te organic and/or inorganic waste products
because of several other existing consl.~Li..ls.
These include:
1. The lack of biologically selected adaptation ( accli.l.~ lion )ofthe microlgani~...s toward
the envhol---lt;..l into wich they will be relarged into.
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2. The too small population of microol~slns in front of such a large amount of work in ratio
with the maximum potential retention time available for the acclinl~ lion and biodegradation
work to occur.
3. The too small counts of microorganisms to carry the beneffts of Bio-Auglllel,lalion ( BOD
reduction and organic matter accum~ tions and solidification ) along the municipal sewage
system.
4. The lack of microorg~nisms natural en_ymatic co-factors and probiotics in the actual
commercial formulations.
5. The abusive costs related to the multiple injections of large number of microorg~nism.~ for
some type of applications makes this technology not profitable for the users.
6. The manpower collsl~illls, since the human manipulation are often required in order to
deliver the right product in the right quantity at the right time. The injection of the bacterias
at a precise moment is a key element for the success of a Bio-treatment.
7. The conslanl microorg~ni~mc washout in the water phase during the day.
8. The pr~sellce of h~digenous ba ;l~lias wich compele with the supplemental Bio-Augmented
bacteria.
SUMMAlRY OF T~ INVENTION
Ther~r~le, it is an object of the present invention to e.limin~te the problems encountered with
the application of the conventionnal technic of Bio-Augnlenlalion and/or Bio-Remediation.
This object is achieved with an efficient and economical system hereina~er called
"acclim~ti7.~tion and amplification system", that will allow high production of high capacity
and specir~-iily microolga~ ,."s for biodegradation of organic and/or inorganic waste products.
In this system, a single inoculum is used for producing huge amounts of bacterias which, in
turn allow multiple il~,e~ ons of a large number of selectively adapted microorg~ni~m.~. Such
has the effect of greatly reducing the cost in material and spare the use of organic solvants,
thus being beneficial to the en~holllnenl and the general population.
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The system according to the invention uses waste water as the cultivation medium for
microorg~ni.cm~. By doing so, it provides an environment that will be optimal for the
amplification of the culture strains of microolg~ ",~ bearing a high capacity and specificity
for organic biodegradation. The system provides a confined and controlled transit environment
for microolg~ which can be located at a distance up to 100 m from the waste water
collector. The acclilll~ ion and amplification process uses waste water as microorganism
cultivation medium by pumping waste water with peristaltic pumps into a pre-acclim~1i7~tion
and amplific~tion chamber (PAAC) and subsequently releasing this medium cont~ining the
;."AIi,.ed microolganJ~,.ns in the effluent. The waste water intàke may be set on a sequential
or continuous basis. Thus, the &lllpL~lion system that is mounted in parallel relationship with
respect to the waste water colleclor allows production of massive quantities of microorg~ni~m~
from a single inoculum. It also allows in;e,~liOIl ofthe microol~al-i~.ns fermentation by-products
(ei~yllles and natural co-factors) in the effluent to be treated since both microorg~ni.cm~ and
cultivation medium are released at the same time. This has the effect of providing to the
microorg~ni.~m~ with all their natural el~y"les and co-factors.
More particularly, the system co~llprises an ~cc~ lion/ampliffcation chamber for producing
massive quantity of rnicroor~ni.~m~. Sul~mel~ible pumps are used for bringing the water from
the ef~uent into the ac~ l;"~A~ lion ch~lll)el. The system also uses other pumps for controlling
the eventual intake of microorganism probiotics (nul~ enls), air and oxygen to promote a
better ~cçl: "~ .AIion environment for the microolg~ ".s. The acçlim~ti7.~tion/amplification
chamber is also linked to a pH and telllpel~ re sensor. Finally, in order to maximize the
amplification of the pre-accli~ .ed microorganism population, the
accl;i,,,.li,AIioll/arnplification chamber comprises polymer devices that are specifically design
to increase the growing surface area for the microor~nism.c. The polymer devices are
preferably plastic rollers that are modular and adaptable in size and in shape in order to be
relatively proportional to the scale and the structure of the system.
As a result, the system provides a confined and controlled transit area allowing high
production of biologically adapted IIL ~olgal~i;,.lls that will degrade effectively the excedentary
organic waste materials found in the effluent.
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The use of the PAAC installed near by a waste water collector or sewage is an efficient
approach for co-l~pen.cnl;n~ the various problems encountered during the use of the
conventionnal Bio-Augmentation technic. As a matter offact, the PAAC allows:
1. an accli,..A~ ion period in the medium co~ g the waste products allowing the
microorg~ni~m.s to select specific biological adaptations before they are injected in the
environment to be treated;
2. an amplification period in the same medium providing a perfect period of retention
time, many times daily. This is to allow the microorg~ni cm.c to grow in a much larger number
in order to face the amount of work to be done in the envhol~lel~l to be treated;
3. a part of the very large count of specifically adapted microorg~nicmc will be washed
down the drains, those microorg~ni~ms will carry the benefits of Bio-Augll,enla~ion ( BOD
reduction, FOG reduction ) along the municipal sewage system and in the environment;
4. the presence of essential natural nullil"el,ls for the microorg~nisms in the water
collection to be treated. When the X liters of medium cO~ g the amplified and acclim~ti7:ed
microorg~ni.smc are re-injected into the envilonl-lelll to be treated, not only are the
microorg~niRm~ better adapted and in much larger number but they are also in presence of
large quantity of their own essential natural en~yllldlic co-factors and pro-biotic that are
usually absent in the actual co-llnlelcial formulations;
5. multiple injections of large number of selectively adapted microorg~nicm.c (produced
from a single inoculum) at reasonable price, making this technology more performant,
plo~ e for the user and more beneficial for the envilonlllelll and the population in general;
6. manpower consl~ are limited to the change of a pail of microorg~nicmc and/or
pro-biotics once in a while at a very speciffc and predictable moment;
7. attenuation of the coll~lalll washout of the microorg~ni.cmC in the water phase of a
waste water treatment plant for example in a grease trap. This is due to the fact that the
ion and ~ lifi~lion Ghamber is a confined and transital environment which is not
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a~ected by the daily water flow. Fulhellllore~ the multiple daily addition and/or the continuous
plopllylactic flow of very large number of microorg~nism~ will compensate for the daily wash
out. Hence, the daily wash out, instead of having a dilution effect with a reduction of the
positive effects of the Bio-Augrnentation, will serve as a transport system to carry the benefits
of Bio-Augl~lenlalion down to the municipal sewage.
8. massive elimin~tion of indigenous bacteria in the private and municipal sewage . The
is due to the fact that the PAAC is equipped with a temperature sensor device that can heat up
the PAAC to up to 80~C for a period of at least 10 minlltes. The elimin~tion of indigenous
bacterias is very illlpOI l~lnl and is another feature of the present invention. The Applicant has
noticed over the years that indigenous bacterias found in the waste water collection such as a
grease trap, are pathogenic and/or opportunistic bacterias. They are, for example,
Ill~lhallogenic or Gram negative bacterias which are responsible of bad odors released in the
environlllenl. These bacterias are not adapted to degrade waste.
The fact th~i the PAAC can partially purify the water from the pathogenic microorg~ni.cmc
before they reach the municipal sewage plant and/or the envirol-lllent is itself another feature
of the present invention. This invention not only provides a way for producing high specificity
and high capacity microolg~ s.~.c for organic and/or inorganic degradation but it also
elimin~tion of a large number of pathogenic microorg~nism.c.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 is a generai representation ofthe pre-accli~ ion and amplification system. The
PAAC is equipped of its controlling devices and are located adjacent to the waste water
collector to be treated
Figure 2 is an upper view of the PAAC which is the transit environment for the
microorE~nicm.s proliferation.
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DESCRIPTION OF THE PREFFERED EMBODIMENTS
As mentionned heleh~at)~ve the invention concellls an acclim~ti7.ation and amplification system
which is intended to be used in parallel to the environment to be treated. By using the waste
water to be treated as a cultivation medium for the microorg~ni.~m~ and by releasing it back
to the envilonlll~llL afcer the amplification period, this system produces microorgAni~ms having
their own essential nutrients when they are released in the environment to be treated.
Furthermore, the accli.,.ali~.Alion/amplification process also allows elimination of a large
number of indigenous and pathogenic microorganisms.
A better undel ~lal~dillg of the present invention will be possible upon reading the following
description. The acclim~ti7~tion /amplification system comprises:
The pre-~cçlimAti7.~tion and amplification chamber
The core ofthe system is the pre-acc'i~ lion and ampliffcation chamber (PMC ) which
is pl~r~l~bly a st~-nlei,s steel box or any other material such as iron, plastic, glass with a total
volume varying from 0.1% up to 20% ofthe total volume ofthe emuent to be treated. The box
can be of any other geometrical form, but is preferably rect~n~ r and can be open from the
top. The size and volume is adjusted according to the nature of the application. The challlbel-
has 5 enllances ( ports with check valves ) in order to be connected with di~erenl control
devices described below and/or for any other conjectural needs required for a better
ac~li...h~ l;on and/or amplification ofthe microolg~ni.~",~. The box has at each e~lle-~ily, a
10 cm inlet and a 10 cm outlet located at the top ends. The chamber can be reto-fit on top,
inside, beside, or under a grease trap or a septic tank in order to be directly connected to it.
Temperature Control and Variations
The PAAC is equipped with a heating device ( watts can vary according to the size of the box)
which can vary and/or sustain the tempe.~ re ofthe Accl;~ alion and amplification medium
from 4~ C to 90~ C for periods of time varying from 5 minutes up to 24 hours day.
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pH Control and Variations
The PAAC is equipped with a pH controller which can control and ,..~;,.l~in the pH of the
acl~l;."~ liol1 and amplification medium from pH 2 up to pH 14 for a period oftime varying
from 5 minutes up to 24 hours day.
Oxygen and air Control and Variations
The PAAC is equipped with an air bubble system which can inject little tiny air bubbles
consisting of pure oxygen or any other required gas from the bottom of the box during a
period of time that can vary from a 5 minutes sequence up to 24 hours a day.
Ambiogene System Microorg~ni.~m.~ Inlet
The PMC has a inlet port equipped with an anti-back flow check valve. This port is dedicated
to the sequential addition of Ambiogene System microolg~l-is.,.s via a peristaltic pump.
Pro-Biotic Inlet
The PAAC has an inlet port eql!ipped with an anti-back flow check valve. This port is
dedicated to the sequential addition of microorg~ni~m.~ pro-biotics such as humic acids and/or
any other microorg~ni.cm~ pro-biotic. The addition of microorg~ni~m~ pro-biotics is done via
a peristaltic pump.
Water Inlet
The PAAC has a water inlet port equipped with an anti-back flow check valve. This port is
dedicated to the sequential addition of tap water in order to dilute the accli~ l;on and
amplification medium that may evei~tually contain lethal concentrations of microorganism
growth inhibitors such as chlorines,quaternary amines or any other anti-biotic.
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Free Inlet
The PAAC has an Ad Lib inlet that may serve in case of specific and/or actually unpredictable
needs that may appear in order to provide the best enviro~ L for the optimal amplification
and accl;l"~ lion ofthe microor~ni~m.~.
Transversal ~rowin~ rollers
The PAAC is p~ ~ly equipped with three ( number of rollers Ad libitum according to the
size and shape of the box ) transversal polymer ( or any other approp~ iate material ) rollers in
order to increase the surface area for the growing of the microol~ is."~ Those rollers are
fixed transversally at the air-water interface, and they can rotate freely with the water flood.
The rollers are made in order to increase the surface availability. This feature, in combination
with the PAAC itself, is considered to be a microolg~ "s amplification system.
Peristaltic pumps
The injection of microorg~ni~m~, Pro-biotic and acid-base for pH control in the PAAC are
done with the help of peristaltic pumps directly controlled and synchl olli~ed by the Plant Line
Controller (PLC ).
Submersible pump
The submersible pump will pump the water from the site to be treated (a grease trap for
~mp'e) up to the pre-acclil--Ali~ ion and amplification chamber. The volume to be pumped
can vary in function with the volume of the effluent to be treated. A ratio varying from 0.1%
up to 20% of the effluent to be treated will be use for calculations and calibration of the
s~lllel~i~lc pump and the FAAC. The submersible pump is equippèd with filters for stopping
any particles larger than l cm coming from the effluent. The submersible pump is controlled
and s~nchl onized by the Plant Line Controller ( PLC ).
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Plant Line Controller
The Plant Line Controller is the " central nervous system " of the system. The PLC is
controlling and synchluni~ g the activation of the ,ul)lllelsil~le pump, the air pump, the
peristaltic pumps, the te",pe,~ re element, the pH controller and/or any other device and/or
events that are required to be done automatically.
The procedure of application of the PAAC co"~l"ises the steps for completing oneamplification cycle. Each cycle being based on a 24 hours period. They are:
1. Filling-up the PAAC with medium which consist of water coming from the waste water
collection to be treated.
2. Killing the indigenous microorg~niC ms contained in this medium by heat.
3. Activation of the air bubble system for the duration of the cycle.
4. Setting optimal pH; the pH of the starting medium is usually acidic.
5. Activation of the Pro-biotic peristaltic pumps for the injection of nutrients in the
~ccli~ lion and amplification medium..
6. Injection of Ambiogene bacterias which will serve as the basic inoculum.
7. ''Pre-c~cclim~ti7.~ion and amplification period" which will allows exponential growth of
microorg~ni .~m.c,
8. Re-fill of the PAAC with new "fresh" medium coming from the waste water collection to
be treated. This step is n~..~.~A~ y for keeping the optimal rate of growth of the microorg~niC ms.
The new intake of fresh medium is for diluting the toxic concenl~ ~lions of inhibitors that may
eventually lead to the killing of the microor~ni.cmc~.
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EXAMPLE 1
Cycle description and objectives of the Pre-Acclim~ti7~tion and ~mplification Device on a 24
hours clock
12H00 AM STEP 1: Filling-up the PAAC with new fresh medium
1. Activation of the SUblllt;l ~ible pump in order to pump XX liters of " grease trap fresh
water " up to the ~tai~ s steel pre-accl;".,.l;,.~l;on and amplification chamber ( PAAC ). The
water comes in by the inlet of the PAAC and the over~ow goes out by the outlet of the PAAC,
the total residual volume of grease trap water is now X liters of growing medium in the
PAAC.
12H05 AM STEP 2: Killing the indigenous bacteria in the new medium
2. Activation of the heating system in order to reach a tempel a~ e of 80 degree Celsius
in the PAAC for period varying from 10 to 20 minutes. The water is heated and l~ ined at
80 degree Celsius for at least ten ( 10 ) minutes in order to destroy most of the indigenous
microorg~niems present in the water. Those usually compete with the Ambiogene
microolganis... during the accli",~li,.AIion and amplification phase.
12H20 STEP 2.1: The setting of the optimal ternperature
2.1 The heating system brings the telnpe~ Jre in the PAAC from 80 degree Celsius, down
to the optimal temperature for the growing and the acclimAIi~.~l;on of the AMBlOGENE
microorg~niem.e Can be set between 4 and 80 Celsius depending on the application.
12H30 AM STEP 3: Provide oxygen in the new growing medium
3 . Activation of the air bubble system for the rest of the cycle. The air bubbles stimulates
the microorg~nieme growth and the aerobic digestion
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12H30 AM STEP 4: O~lh,l;ze the pH that is usually acidic
4. Activation of the pH controller in order to eslablisll the optimal pH. A pH of
applu~ ately 7 will be optimal for the growth ofthe Ambiogene microolg~ni~.,.s
12H30 AM STEP 5: Inject nutrients to oplinli~e the bacteria metabolism
5 . Activation of the Pro-biotic peli~dlLic pumps in order to inject X ml of Pro-Biotic.The
addition of Pro-Biotic is facultative but, it usually helps the microorg~ni~m.c to grow faster.
12H30 AM STEP 6: Inject Ambiogene bacteria's
6. Activation of the peristaltic pump in order to inject X ml of BIO-FILM. The addition
of the AMBIOGENE MICROORGANISMS will serve as a basic inoculum to start the
gr()wing phase. The volume ofthe AMBIOGENE MICROORGANISMS innoculum can vary
in with the size of the application
FROM 12H30 TO 6H00 AM STEP 7: Pre-Acclim~ti7~tion & Amplification
7. This period is called the " Pre-acclim~ti7~tion and Amplification period " . It is during
this period of time that the AMBIOGENE microolg~ will biologically and
morphologically develop adaptations in order to adapt their metabolism to the physico-
chel~L -1 composition ofthe m~ m where they will have to evolve ( and work ) later on. The
AMBIOGENE microorg~ni~ms will e"ponenlially multiply on a logaliLl~n~c basis. It is also
during this period oftime that the AMBIOGENE microolg~l-isl~ will secrete in their growing
medium, all the necess&ly el~y---es, natural co-factors and other essçnti~l Bio-molecules that
will help them to evolve ( and work ) at their maximal potential in their future working
environment.
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6H00 AM STEP 1: Re-Filling up the PMC with XX liters of new fresh medium
( mode: PAAC Empty or Full )
8a. Re-activation of thé su~ el ~ le pump in order to re-pump up XX liters of " fresh"
liquid into the Pre-~ccli".~li7.~lion and amplification chamber ( PAAC ) . The re-activation of
the submersible pump as a double impact:
a. It brings up to the PAAC ( via the Inlet ) XX liters of new liquid medium in order
to re-start a new cycle with new AMBIOGENE microorg~nisms
b. It pushes out ofthe PAAC down to en~,irolllllellt to be treated, zillions of specifically
ed AMBIOGENE microor~ni~ms plus X liters of medium fully loaded with natural
el~ymes, co-factors and essential Bio-molecules already available for the AMBIOGENE
microolgani~ 3 that will enter almost immediately in a new glUWing and biodegrading phase
in the grease trap and/or in any other envil olllllelll to be treated.
OR
6H00 AM STEP 1: Re-filling up the PAAC with X ml of new fresh medium.
( Mode: PAAC Feed Batch )
8b. The Re-activation of the submersible pump in order to re-pump only 1% of the total
volume of the PAAC with " fresh " new liquid coming up from the effluent to be treated can
also be done on a every x ml per minutes or hour basis. In this case, the re-activation of the
Sl~ le pump will transform the PAAD in a prophylactic system, treating the effluent on
a on going basis ( Feed Batch ). In this mode the re-activation of the subnlel ~ible pump will
have the same double impact as mention in 8a.
9. Repetition of steps 2,3,4,5,6,7, 8a or 8b... 1,2.3,4,5,6,7,8, 8aor 8b...and so on.
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As a result of this cycle, it is now possible to inject daily, the equivalent of Zillions in-situ
selectively adapted microo.gal1ism in a volume of approximately X liters, for the price of a
single injection of few hundreds ml per day, usually cont~ining only millions of non specific
and non acclim~ti~ed bacteria's. It is atso possible to treat in a prophylactic manner an
effluent that may need this kind of treatment.
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