THE USE OF OXIDIZING AND NON-OXIDIZING BIOCIDES FOR
CONTROL OF BACTERIA TOLERANT TO STABILIZED-OXIDANT
TREATMENT
Background of the Invention
The present invention relates generally to compositions of matter,
apparatuses and methods useful in detecting, identifying, and addressing
microorganisms present in commercial process systems.
The presence and growth of certain microorganism in commercial
process systems is an ongoing challenge. Many of the various stages of commercial
process systems contain a variety of conditions having different amounts of water,
nutrients, heat, shelter, anchoring substrates, chemical conditions, and/or an absence
of predators which often function as environmental niches suitable for colonization
by all sorts of microorganisms. Population growth by these microorganisms often
poses a number of problems including degrading process functions and fouling the
end products.
One such problem is microorganism induced crust deposit formation.
Crust is the accumulation on a surface of an item present in a commercial process
system of a rigid solid composition comprising deposited organic and/or inorganic
material. The crust can be secretions and/or colonies of microorganisms themselves.
In particular crust can include or consist of the accumulation of one or more kinds of
hard shelled and/or chitin bearing and/or coral organisms. Crust can have many
negative impacts on systems such as decreased operational efficiency, premature
equipment failure, loss in productivity, loss in product quality, and increased healthrelated
risks. Worst of all crust must often be physically removed by scraping or
other physical means and this requires expensive shut downs or disassembly of part
or all of the process system.
Another problem microorganisms pose is through the formation of
biofilms. Biofilms are layers of organic materials comprising microorganisms or
exopolymeric substance secreted by microorganisms which aid in the formation of
microbial communities. Biofilms can grow on the surfaces of process equipment as
well as in pools of fluid. These biofilms are complex ecosystems that establish a
means for concentrating nutrients and offer protection for growth. Biofilms can
accelerate crust, corrosion, and other fouling processes. Not only do biofilms
contribute to reduction of system efficiencies, but they also provide an excellent
environment for microbial proliferation of other microorganisms including
pathogenic organisms. It is therefore important that biofilms and other fouling
processes be reduced to the greatest extent possible to maximize process efficiency
and minimize the health-related risks from such pathogens.
Several factors contribute to the extent of biological contamination
and govern the appropriate response. Water temperature; water pH; organic and
inorganic nutrients, growth conditions such as aerobic or anaerobic conditions, and
in some cases the presence or absence of sunlight, etc. can play an important role.
These factors also help in deciding what types of microorganisms might be present
in the water system and how best to control those microorganisms. Proper
identification of the microorganism is also crucial to responding appropriately.
Differences regarding whether the microorganisms are plants, animals, or fungi, or if
they are planktonic or sessile determines how effective various biocontrol strategies
will be. Because different microorganisms induce different problems, proper
identification is crucial to properly remediating unwanted microbial effects. Finally
because chemically caused problems cannot be remediated with biocides, it is also
necessary to identify which problems have non-biologically based origins.
One category of matter commonly used to respond to micro-organism
infestations is oxidants. Oxidants, such as sodium hypochlorite, are highly reactive
and effectively "burn" away the cell walls of many microorganisms. Unfortunately
because they are so reactive such oxidants often either lose effectiveness very
quickly or they corrode or otherwise interact harmfully with other components or
materials used in commercial process systems.
As a result, a number of technologies have been developed to
stabilize oxidants. Some methods are described in US Patents 5,565,109 and
7,776,363. Such stabilization results in a countering of the so called oxidant demand
effect. In an oxidant demand effect reaction, because the oxidant is in the presence
of something that it is highly reactive with, the oxidant tends to rapidly react and
become unavailable for use as a biocide. By stabilizing oxidants, the oxidant
remains present in the system for a longer period of time and is capable of
suppressing microorganisms for an extended period of time.
In the biological world, however the demise of one organism often
means a niche becomes available for another organism (which was previously
suppressed by its now dead neighbors) to colonize. This in fact is often the case in
process water treated with stabilized oxidant biocides. Many organisms (such as for
example Sphingomonas sp., Acinetobacter, and Flavobacterium) secrete chemicals
which can destroy the oxidant stabilizers and once their former competitors are
killed off by the stabilized oxidants, they are capable of colonizing those
environments despite the presence of the stabilized oxidants. As a result methods
and apparatuses are needed to follow up after treating a commercial process system
with a stabilized oxidant biocide to ensure these organisms have been eradicated.
Thus it is clear that there is clear utility in novel methods and
compositions for the follow up to stabilized oxidant biocide treatment of a
commercial process system. The art described in this section is not intended to
constitute an admission that any patent, publication or other information referred to
herein is "Prior Art" with respect to this invention, unless specifically designated as
such. In addition, this section should not be construed to mean that a search has been
made or that no other pertinent information as defined in 37 CFR § 1.56(a) exists.
Brief Summary of the Invention
At least one embodiment of the invention is directed towards a
method of addressing a microorganism infestation in a process water system. The
method comprises the steps of: introducing a biocidal composition comprising an
oxidant and an oxidant stabilizer into the process water system, the stabilizer
allowing the oxidant to persist as a biocide despite the high demand for the oxidant
in the system, detecting the presence of an organism capable of degrading the
oxidant stabilizer, and if so detected, introducing a composition of matter capable of
neutralizing the degrading organism without otherwise impairing the effectiveness
of the biocidal composition.
The stabilizer may comprise a nitrogen-based compound. The
organism may be a urease secreting organism and/or a nitrifying organism. The
composition may be susceptible to high demand. The organism may remain
neutralized long after the composition susceptible to high demand has been
completely consumed. The detection may be accomplished by at least one item
selected from the list consisting of DNA analysis, PCR analysis, qPCR analysis,
urease detection, ammonia detection, ammonia monooxygenase detection, nitrite
oxidoreductase, nitrate detection, nitrite detection, hydroxylamine detection, and any
combination thereof. But for the introduction of the neutralizing composition, the
organism may actually better thrive in the presence of the biocidal composition than
in its absence because it feeds on the stabilizer or a derivative thereof. The
neutralizing composition may comprise Glutaraldehyde, DBNPA, sodium
hypochlorite, inorganic chloramine, and any combination thereof. The neutralizing
composition may comprise inorganic chloramine if a urease-degrading population is
detected.
Detailed Description of the Invention
The following definitions are provided to determine how terms used
in this application, and in particular how the claims, are to be construed. The
organization of the definitions is for convenience only and is not intended to limit
any of the definitions to any particular category.
"Adaptor" means an organism that exhibits some level of tolerance
to the biocontrol program. When the adaptor's microbial competition is reduced by
a biocide, this adaptive organism is able to flourish and may form a biofilm.
"Biological" means a composition of matter in which at least 10% of
the composition (by volume or mass) comprises cells from an organism.
"High Demand" or "High oxidant demand" means the presence of
a chemical that is highly reactive with oxidants in a particular environment and the
oxidant will therefore rapidly react and will not persist for any appreciable extent
after a short period of time. High demand conditions deplete unstabilized oxidants
more rapidly than stabilized oxidants.
"Opportunist" means an organism that thrives by settling into preestablished
biofilms, crusts, deposits, or other colonies of organisms, and tends to
supplant, displace, or coexist alongside pioneer organisms and/or previous
opportunist organisms.
"PCR Analysis" means polymerase chain reaction analysis.
"Probe" means a composition of matter constructed and arranged to
bind to a targeted section of DNA and which can be readily detected when so bound
and thereby be used to indicate the presence or absence of the targeted section of
DNA.
"qPCR Analysis" means quantitative polymerase chain reaction
analysis.
"Microorganisms" mean any organism small enough to insinuate
itself within, adjacent to, on top of, or attached to equipment used in an industrial
process (including papermaking), it includes but is not limited to those organisms so
small that they cannot be seen without the aid of a microscope, collections or
colonies of such small organisms that can be seen by the naked eye but which
comprise a number of individual organisms that are too small to be seen by the
naked eye, as well as one or more organisms that can be seen by the naked eye, it
includes but is not limited to any organism whose presence, in some way impairs the
industrial process such as forming plugs in nozzles and/or within felts and/or
causing defects within paper sheets.
In the event that the above definitions or a description stated
elsewhere in this application is inconsistent with a meaning (explicit or implicit)
which is commonly used, in a dictionary, or stated in a source incorporated by
reference into this application, the application and the claim terms in particular are
understood to be construed according to the definition or description in this
application, and not according to the common definition, dictionary definition, or the
definition that was incorporated by reference. In light of the above, in the event that
a term can only be understood if it is construed by a dictionary, if the term is defined
by the Kirk-Othmer Encyclopedia of Chemical Technology, 5th Edition, (2005),
(Published by Wiley, John & Sons, Inc.) this definition shall control how the term is
to be defined in the claims.
Oxidants (such as sodium hypochlorite) are routinely applied to
industrial process systems (such as paper systems) to control microbial growth and
deposit formation. In many instances they include a halogen (such as chlorine)
which is often consumed due the presence of high demand compositions (e.g.
sulfite) in many environments and is therefore often stabilized with stabilizing
compounds (such as nitrogenous compounds) to enhance persistence of the halogen
residual in water systems with high halogen-demand and to improve compatibility
with process additives (e.g. optical brightening agents, dyes, strength aids, and
sizing agents) relative to free or unstabilized-chlorine.
As described in US Patent 7,773,363, one composition used to
stabilize oxidants such as chlorine is urea. Unfortunately, in some situations it has
been observed that applications of urea-stabilized chlorine programs over time
resulted in the high persistence of certain bacteria and fungi. This persistence may
occur despite increased treatment levels. This is consistent with a failure of the
oxidant stabilizer to continue to effectively stabilize the oxidant leading to more
rapid consumption of the oxidant and less effective control of microbial growth.
Closer observation indicated that the bacteria were secreting chemicals (such as
enzymes) which degraded the stabilizer and resulted in the loss of halogen due to
halogen demand and increased tolerance of the organisms to oxidant treatment.
In at least one embodiment after implementing a stabilized oxidant
biocide regimen, a post treatment strategy is implemented to address the rise of
organisms that degrade the oxidant stabilizer. In at least one embodiment the
strategy provides for preventing or controlling the growth of urea-degrading
organisms. This allows urea-stabilized chlorine programs to perform at their
intended level and prevents the need for increased chlorine usage in an attempt to
maintain control of these microbial populations and reduce the chlorine-demand
created by their enzyme byproducts.
In at least one embodiment the post treatment strategy involves the
use of compositions of matter which do not react with the stabilized oxidant or
oxidant stabilizer and which also are effective at killing the stabilizer-degrading
organisms. In at least one embodiment the composition of matter is a non-oxidizing
biocide such as but not limited to dibromonitrilopropionamide (DBNPA) (for
example as found in product Nalcon 7649, sold by Nalco Company, Naperville, IL)
and glutaraldehyde (for example as found in product Nalcon 7634 sold by Nalco
Company, Naperville, IL). In at least one embodiment sodium hypochlorite or
inorganic chloramine programs generated by blending sodium hypochlorite with
ammonium salts (e.g. ammonium sulfate or ammonium bromide) are also effective
for the control of a stabilizer degrading organisms such as those that that exhibit
tolerance to urea-stabilized chlorine programs.
In at least one embodiment the method involves the step of
anticipating the rise of a stabilizer degrading organisms before it degrades the
stabilizer (or at least before it does so to a detectable or significant degree) and
neutralizing the organism before the unwanted degradation occurs. For example any
one of the methods, compositions, and apparatuses for detecting a diversity index
described in US Patent Application 13/550,748 could be used to anticipate the rise
of stabilizer degrading organisms. As a result the post treatment strategy could
involve determining if a stabilizer degrading organism would arise and after it has
been so determined but before the organisms has actually significantly degraded the
stabilizer would biocides targeting that organisms be applied.
In at least one embodiment the composition added to kill the
stabilizer-degrading organisms is a composition susceptible to high demand.
Compositions which are susceptible to high demand within a given environment, by
definition will not persist long in that environment. As a result, it would not be
expected that their application could impart long term anti-microbial benefits.
However when applied as a post treatment to a stabilized biocide, the secondary
oxidant, which is susceptible to high demand kills off opportunistic adaptor
organisms which degrade the stabilizer, and with the stabilizer no longer degraded,
the stabilized oxidant continues to maintain the process system in an acceptable
antimicrobial state. In at least one embodiment the post treatment of a secondary
oxidant maintains the absence of stabilized oxidant resistant organisms long after the
composition susceptible to high demand is completely consumed by reaction or
diluted from the process system. In at least one embodiment the composition
susceptible to high demand is sodium hypochlorite or inorganic monochloramine.
In at least one embodiment the stabilizer degrading organism is an
organism capable of degrading urea or is a nitrifying organism. As described for
example in scientific papers: Bacterial Nitrification in Chloraminated Water
Supplies, by David A. Cunliffe, Applied And Environmental Microbiology, Vol. 57,
No. 11, Nov. 1991, pp. 3399-3402, Physiological Studies of Chloramine Resistance
Developed by Klebsiella pneumonia under Low-Nutrient Growth Conditions , by
Mic H. Stewart et al., Applied And Environmental Microbiology, Vol. 58, No. 9,
Sept. 1992, pp. 2918-2927, and Isolation from Agricultural Soil and
Characterization of a Shingomonas sp. Able to Mineralize the Phenyl Urea
Herbicide Isoproturon , by Sebastian R. S0rensen, et al., Applied And Environmental
Microbiology, Vol. 67, No. 12, Dec. 2001, pp. 5403-5409, nitrifying and ureadegrading
organisms derive energy by oxidizing nitrogen containing compounds
(such as ammonia or urea) into ammonia, nitrites, or nitrates. As a result these
organisms not only degrade nitrogen based stabilizers but may actually better thrive
in the presence of a nitrogen based stabilizer biocide than in its absence.
Furthermore, nitrite accelerates the decay of inorganic chloramine residuals.
In at least one embodiment the post treatment includes the step of
detecting the presence of a urea-degrading organism and applying a response to
neutralize the urea-dedgrading organism without otherwise harming the stabilized
oxidant composition. In at least one embodiment the detection is achieved through
DNA based analysis involving the use of PCR primers to detect the presence and/or
absence and quantity of urea-dedgrading organisms. US Patent 5,928,875 describes
the use of PCR primers to detect the presence or absence of spore forming bacteria.
In at least one embodiment the primer is targeted towards a part of a DNA strand
which is highly conserved among urea-dedgrading organisms. In at least one
embodiment the PCR analysis involves utilizing one or more of the methods
described in the Article Primer Directed Enzymatic Amplification of DNA with a
Thermostable DNA Polymerase , by Randall Saiki et al., Science, Volume 239, pp.
487-491 (1988). In at least one embodiment the PCR analysis involves utilizing one
or more of the methods described in the Article Specific Synthesis of DNA in Vitro
via a Polymerase-Catalyzed Chain Reaction, by Kary Mullis et al., Methods In
Enzymology, Volume 155, pp. 335-350 (1987).
In at least one embodiment the post treatment includes the step of
detecting the presence of a nitrifying organism and applying a response to neutralize
the nitrifying organism without otherwise harming the stabilized oxidant
composition. In at least one embodiment the detection is achieved through DNA
based analysis involving the use of PCR primers to detect the presence and/or
absence and quantity of nitrifying organisms. US Patent 5,928,875 describes the use
of PCR primers to detect the presence or absence of spore forming bacteria. In at
least one embodiment the primer is targeted towards a part of a DNA strand which is
highly conserved among nitrifying organisms. In at least one embodiment the PCR
analysis involves utilizing one or more of the methods described in the Article
Primer Directed Enzymatic Amplification of DNA with a Thermostable DNA
Polymerase , by Randall Saiki et al., Science, Volume 239, pp. 487-491 (1988). In at
least one embodiment the PCR analysis involves utilizing one or more of the
methods described in the Article Specific Synthesis of DNA in Vitro via a
Polymerase-Catalyzed Chain Reaction, by Kary Mullis et al., Methods In
Enzymology, Volume 155, pp. 335-350 (1987).
In at least one embodiment the PCR analysis is a qPCR analysis as
described in Trade Brochure qPCR guide, prefaced by Jo Vandesompele, (as
downloaded from website http://www.eurogentec.com/file-browser.html on January
19, 2012). In at least one embodiment the method is a quantitative qPCR analysis.
In at least one embodiment the method is a qualitative qPCR analysis.
In at least one embodiment, the polymerase chain reaction (PCR) is a
method for targeting sequences of nucleic acid (DNA or RNA) and increasing the
copy number of the target sequence to obtain useful quantities of nucleic acid for
down-stream analysis. This method can be applied to the detection of ureadegrading
and nitrifying organisms in a variety of samples that include, but are not
limited to, machine felts, sheet defects, machine deposits, etc.
In at least one embodiment, once DNA is extracted from the sample,
using any of the DNA extraction kits available commercially, it can be analyzed in
real-time using a PCR approach such as a Quantitative PCR approach. Quantitative
PCR utilizes the same methodology as PCR, but it includes a real-time quantitative
component. In this technique, primers are used to target a DNA sequence of interest
based on the identity of the organism or function of a specific gene. Some form of
detection such as fluorescence may be used to detect the resulting DNA or 'DNA
amplicon'. The change in fluorescence is directly proportional to the quantity of
target DNA. The number of cycles required to reach the pre-determined
fluorescence threshold is compared to a standard that corresponds to the specific
DNA target. A standard is typically the target gene that is pure and of known
quantity at concentrations that span several logs. The number of copies of target
DNA present in the sample is calculated using the standard curve. The copy number
per sample is then used to determine the number of cells per sample.
In at least one embodiment a primer set is used which targets DNA
sequences from nitrifying organisms using a conservative approach to quantify total
organisms. In at least one embodiment a primer set is used which targets ureadegrading
organisms, including, but not limited to, Sphingomonas sp.,
Sphingomonas spp. , Acinetobacter, and Flavobacterium. In at least one
embodiment the primer is used to distinguish between urea-degrading and non-ureadegrading
organisms.
In at least one embodiment a primer set is used which targets DNA
sequences from nitrifying organisms using a conservative approach to quantify total
organisms. In at least one embodiment a primer set is used which targets nitrifying
organisms, including, but not limited to, Nitrosomonas, Nitrosolobus,
Nitrosococcus, Nitrosovibrio, Niotrosospira, Nitrobacter, and Nitrococcus . In at
least one embodiment the primer is used to distinguish between nitrifying and nonnitrifying
organisms.
In at least one embodiment the detection method involves detecting
the telltale presence of a stabilizer degrading organism. For example because many
of these organisms secret urease so the method involves detecting the presence of
urease or ammonia. Also many nitrifying organisms convert the stabilizer into end
reactants that would otherwise not result from the oxidation reaction of the biocide
so the method involves detecting the presence of these end reactants (such as the
specific nitrates, hydroxylamine, or nitrites produced by their digestion of
nitrogenous compounds). In addition the strategy could involve detecting other nonnitrogen
based end reactants that these organisms produce (e.g. ammonia
monooxygenase or nitrite oxidoreductase). Finally another strategy could involve
noting that if when using a diversity index analysis, if the overall population grows
in response to applying a stabilized oxidant biocide, but it briefly declines in
response to applying a non-stabilized version of that biocide, it suggests the
colonization of the system by organisms feeding on the stabilizer. Another strategy
could involve noting the development of the population by screening the
antimicrobial effect of the stabilized oxidant in a laboratory.
EXAMPLES
The foregoing may be better understood by reference to the following
examples, which are presented for purposes of illustration and are not intended to
limit the scope of the invention.
The process water stream of a paper mill was treated using a ureastabilized
chlorine program. While initial laboratory screening prior to the
application of stabilized-oxidant program indicated that urea-stabilized chlorine
programs were effective in controlling microbial growth in this mill, repeat
screening indicated that this program was no longer effective (Table 1). This
bacterial population also exhibited tolerance to isothiazolone biocides.
Table 1 show that a Laboratory screening on process water treated
with NaOCl was compared to screening results on a sample of process water after
10-months of treatment with urea-stabilized chlorine. Results indicate the
development of a microbial population tolerant to process water treatment with a
urea- stabilized chlorine program. Process water samples were challenged with
untreated process water (1% v/v) after the 4-hours samples were plated. Typically,
urea- stabilized chlorine exhibits excellent long-term preservation with low bacterial
densities at 4- and 24-hours relative to an untreated control. In Table 1, preservation
from urea- stabilized chlorine was observed prior to the application of this program
to the mill water system and development of the tolerant population. Repeat
screening demonstrated the tolerance of the new mill population to the ureastabilized
chlorine program.
Table 1:
Several bacteria surviving C12/60615 treatment exhibited yellow or
orange pigmentation when samples were plated on non-selective agar media. These
organisms were isolated into pure culture and were identified as Sphingomonas
species based on DNA sequencing. These isolates exhibited the ability to produce
urease. Urease enzyme breaks down urea into ammonia, which can then provide a
nutrient source for other bacteria. Furthermore, urease has been determined to serve
as a significant source of chlorine-demand and also exhibits some interaction with
urea- stabilized chlorine based on higher ammonia levels after urea- stabilized
chlorine is exposed to the urease enzyme (Tables 2 and 3). This indicates the
degradation of urea, which likely impacts the performance of urea- stabilized
chlorine programs.
Table 2: Urease enzyme leads to a reduction in chlorine residuals,
indicating that this enzyme serves as a significant source of chlorine demand.
Table 3: Exposure of urea- stabilized chlorine to urease led to an increase in
ammonia concentrations (NH3-N). This indicates the degradation of urea,
which likely impacts the performance of urea- stabilized chlorine programs.
This problem of increased chlorine-demand and the development of a
tolerant population appeared to be specific to applications using urea- stabilized
chlorine and has not been observed in applications using dimethyl hydantoinstabilized
chlorine programs or inorganic chloramine programs generated by
blending sodium hypochlorite with ammonium salts. Systems treated with
inorganic chloramine are more likely to develop tolerance issues related to the
development of nitrifying populations of bacteria.
Antimicrobial screening of non-oxidizing biocides indicated that
dibromonitrilopropionamide and glutaraldehyde provided effective control of
urease-producing microorganisms and are compatible with urea-stabilized chlorine.
Sodium hypochlorite or inorganic chloramine programs generated by blending
sodium hypochlorite with ammonium salts (e.g. ammonium sulfate or ammonium
bromide) are also effective for the control of urease-producing bacteria that
exhibited tolerance to urea- stabilized chlorine programs (Tables 4, 5 and 6).
Table 4: Glutaraldehyde, Dibromonitrilopropionamide, and NaOCl are
particularly effective at controlling C12-urea tolerant bacteria at typical use
rates.
Table 5 illustrates laboratory screening was conducted using process
water treated with urea-stabilized chlorine. Process water samples were challenged
with untreated process water (1% v/v) after the 4-hour samples were plated. Results
indicate the development of a microbial population tolerant to treatment with a ureastabilized
chlorine program, as this program had little effect in reducing bacterial
density relative to an untreated control. Glutaraldehyde,
Dibromonitrilopropionamide, , and inorganic chloramine prepared by blending
NaOCl and ammonium sulfate were particularly effective at controlling C12-urea
tolerant bacteria at typical use rates. NaOCl was also effective, but was required at
higher concentrations, which could lead to incompatibility with other additives used
in the papermaking process. Glutaraldehyde and dibromonitrilopropionamide were
compatible with C12-urea. Isothiazolone and bronopol were not effective at
controlling the tolerant population.
Table 5:
The result of these examples demonstrates that stabilized oxidant
biocide treatments may cease to be effective if there are stabilizer degrading
organisms present in the system and in order to re-establish the effectiveness of the
stabilized oxidant biocide treatment the degrading organisms must be neutralized in
a manner that does not otherwise impair or degrade the stabilized oxidant biocide or
any other material or item also present in the process water system.
While this invention may be embodied in many different forms, there
described in detail herein specific preferred embodiments of the invention. The
present disclosure is an exemplification of the principles of the invention and is not
intended to limit the invention to the particular embodiments illustrated. All patents,
patent applications, scientific papers, and any other referenced materials mentioned
herein are incorporated by reference in their entirety. Furthermore, the invention
encompasses any possible combination of some or all of the various embodiments
described herein and/or incorporated herein. In addition the invention encompasses
any possible combination that also specifically excludes any one or some of the
various embodiments described herein and/or incorporated herein.
The above disclosure is intended to be illustrative and not exhaustive.
This description will suggest many variations and alternatives to one of ordinary
skill in this art. All these alternatives and variations are intended to be included
within the scope of the claims where the term "comprising" means "including, but
not limited to". Those familiar with the art may recognize other equivalents to the
specific embodiments described herein which equivalents are also intended to be
encompassed by the claims.
All ranges and parameters disclosed herein are understood to
encompass any and all subranges subsumed therein, and every number between the
endpoints. For example, a stated range of " 1 to 10" should be considered to include
any and all subranges between (and inclusive of) the minimum value of 1 and the
maximum value of 10; that is, all subranges beginning with a minimum value of 1 or
more, (e.g. 1 to 6.1), and ending with a maximum value of 10 or less, (e.g. 2.3 to
9.4, 3 to 8, 4 to 7), and finally to each number 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10
contained within the range.
This completes the description of the preferred and alternate
embodiments of the invention. Those skilled in the art may recognize other
equivalents to the specific embodiment described herein which equivalents are
intended to be encompassed by the claims attached hereto.

Claims
What is claimed is:
1. A method of addressing a microorganism infestation in a process water
system, the method comprising the steps of:
introducing a biocidal composition comprising an oxidant and an oxidant
stabilizer into the process water system, the stabilizer allowing the oxidant to persist
as a biocide despite the high demand for the oxidant in the system,
detecting the presence of an organism capable of degrading the oxidant
stabilizer,
if so detected, introducing a composition of matter capable of neutralizing
the degrading organism without otherwise impairing the effectiveness of the biocidal
composition.
2. The method of claim 1 in which the stabilizer comprises a nitrogen-based
compound.
3. The method of claim 1 in which the organism is a urease secreting organism.
4. The method of claim 1 in which the organism is a nitrifying organism.
5. The method of claim 1 in which the composition is susceptible to high
demand and the organism remains neutralized long after the composition susceptible
to high demand has been completely consumed.
6. The method of claim 1 in which the detection is accomplished by at least one
item selected from the list consisting of DNA analysis, PCR analysis, qPCR
analysis, urease detection, ammonia detection, ammonia monooxygenase detection,
nitrite oxidoreductase, nitrate detection, nitrite detection, hydroxylamine detection,
and any combination thereof.
7. The method of claim 1 in which but for the introduction of the neutralizing
composition, the organism would actually better thrive in the presence of the
biocidal composition than in its absence because it feeds on the stabilizer.
8. The method of claim 1 in which the neutralizing composition comprises
Glutaraldehyde, DBNPA, and any combination thereof.
9. The method of claim 1 in which the neutralizing composition comprises
sodium hypochlorite.
10. The method of claim 1 in which the neutralizing composition comprises
inorganic chloramine if a urease-degrading population is detected.
11. The method of claim 1 in which the detection is accomplished by detecting
the presence of compounds in the process water that are typical of what nitrifying
organisms convert the stabilizer into and which are compounds that would not
otherwise not result from the oxidation reaction of the biocide.
12. The method of claim 1 in which the detection is accomplished by detecting
the presence of compounds in the process water that are typical of what ureadegrading
organisms convert the stabilizer into and which are compounds that would
not otherwise not result from the oxidation reaction of the biocide.