# Patent

## Title
Compositions of dibromomalonamide and their use as biocides

## Abstract

A biocidal composition comprising 2,2-dibromomalonamide and an aldehyde-based biocidal compound, and its use for the control of microorganisms in aqueous and water-containing systems.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims the benefit of U.S. Provisional Application Ser. No. 61/246,186, filed Sep. 28, 2009, which is incorporated herein by reference in its entirety.


FIELD OF THE INVENTION
The invention relates to biocidal compositions and methods of their use for the control of microorganisms in aqueous and water-containing systems. The compositions comprise 2,2-dibromomalonamide and an aldehyde-based biocidal compound.
BACKGROUND OF THE INVENTION
Water systems provide fertile breeding grounds for algae, bacteria, viruses, and fungi some of which can be pathogenic. Microbial contamination can create a variety of problems, including aesthetic unpleasantries such as slimy green water, serious health risks such as fungal, bacterial, or viral infections, and mechanical problems including plugging, corrosion of equipment, and reduction of heat transfer.
Biocides are commonly used to disinfect and control the growth of microorganisms in aqueous and water containing systems. However, not all biocides are effective against a wide range of microorganisms and/or temperatures, and some are incompatible with other chemical treatment additives. In addition, some biocides do not provide microbial control over long enough time periods.
While some of these shortcomings can be overcome through use of larger amounts of the biocide, this option creates its own problems, including increased cost, increased waste, and increased likelihood that the biocide will interfere with the desirable properties of the treated medium. In addition, even with use of larger amounts of the biocide, many commercial biocidal compounds cannot provide effective control due to weak activity against certain types of microorganisms or resistance of the microorganisms to those compounds.
It would be a significant advance in the art to provide biocide compositions for treatment of water systems that yield one or more of the following advantages: increased efficacy at lower concentrations, compatibility with physical conditions and other additives in the treated medium, effectiveness against a broad spectrum of microorganisms, and/or ability to provide both short term and long term control of microorganisms.
BRIEF SUMMARY OF THE INVENTION
In one aspect, the invention provides a biocidal composition. The composition is useful for controlling microorganisms in aqueous or water containing systems. The composition comprises: 2,2-dibromomalonamide and an aldehyde-based biocidal compound selected from the group consisting of glutaraldehyde, tris(hydroxymethyl)nitromethane, 4,4-dimethyloxazolidine, 7-ethyl bicyclooxazolidine, 1-(3-chloroallyl)-3,5,7-triaza-1-azoniaadamantane chloride, and 1,3,5-triethylhexahydro-s-triazine.
In a second aspect, the invention provides a method for controlling microorganisms in aqueous or water containing systems. The method comprises treating the system with an effective amount of a biocidal composition as described herein.


DETAILED DESCRIPTION OF THE INVENTION
As noted above, the invention provides a biocidal composition and methods of using it in the control of microorganisms. The composition comprises: 2,2-dibromomalonamide and an aldehyde-based (i.e., aldehyde-containing or aldehyde-releasing) biocidal compound selected from the group consisting of glutaraldehyde, tris(hydroxymethyl)nitromethane, 4,4-dimethyloxazolidine, 7-ethyl bicyclooxazolidine, 1-(3-chloroallyl)-3,5,7-triaza-1-azoniaadamantane chloride, and 1,3,5-triethylhexahydro-s-triazine. It has surprisingly been discovered that combinations of 2,2-dibromomalonamide and an aldehyde-based biocidal compound as described herein, at certain weight ratios, are synergistic when used for microorganism control in aqueous or water containing media. That is, the combined materials result in improved biocidal properties than would otherwise be expected based on their individual performance. The synergy permits reduced amounts of the materials to be used to achieve the desired biocidal performance, thus reducing problems caused by growth of microorganisms in industrial process waters while potentially reducing environmental impact and materials cost.
For the purposes of this specification, the meaning of “microorganism” includes, but is not limited to, bacteria, fungi, algae, and viruses. The words “control” and “controlling” should be broadly construed to include within their meaning, and without being limited thereto, inhibiting the growth or propagation of microorganisms, killing microorganisms, disinfection, and/or preservation. In some preferred embodiments, “control” and “controlling” mean inhibiting the growth or propagation of microorganisms. In further embodiments, “control” and “controlling” mean the killing of microorganisms.
The term “2,2-dibromomalonamide” refers to a compound represented by the following chemical formula:




2,2-Dibromomalonamide and the aldehyde-based biocidal compounds of the invention are commercially available and/or can be readily prepared by those skilled in the art using well known techniques. The 1-(3-chloroallyl)-3,5,7-triaza-1-azoniaadamantane chloride compound may be the cis isomer, the trans isomer, or a mixture of cis and trans isomers. Preferably, it is the cis isomer or a mixture of the cis and trans isomers.
In some embodiments of the invention, the weight ratio of 2,2-dibromomalonamide to the aldehyde-based biocidal compound is between about 100:1 and about 1:1500.
In some embodiments of the invention, the weight ratio of 2,2-dibromomalonamide to the aldehyde-based biocidal compound is between about 100:1 and about 1:400.
In some embodiments, the weight ratio of 2,2-dibromomalonamide to the aldehyde-based biocidal compound is between about 50:1 and about 1:350.
In some embodiments, the weight ratio of 2,2-dibromomalonamide to the aldehyde-based biocidal compound is between about 9:1 and about 1:320.
In some embodiments, the aldehyde-based biocidal compound is glutaraldehyde and the weight ratio of 2,2-dibromomalonamide to glutaraldehyde is from about 20:1 to about 1:20, alternatively from about 9:1 to about 1:9, or alternatively from about 9:1 to about 1:1. In some embodiments the weight ratio is from about 2:1 to about 1:350, alternatively from about 1:1 to about 1:330, or alternatively from about 1:5 to about 1:320. In some embodiments, the weight ratio is from about 20:1 to about 1:400, alternatively from about 10:1 to about 1:350, or alternatively from about 9:1 to about 1:320.
In some embodiments, the aldehyde-based biocidal compound is tris(hydroxymethyl)nitromethane and the weight ratio of 2,2-dibromomalonamide to tris(hydroxymethyl)nitromethane is from about 1:1 to about 1:50, alternatively from about 1:2 to about 1:40, or alternatively from about 1:2.5 to about 1:40.
In some embodiments, the aldehyde-based biocidal compound is 7-ethyl bicyclooxazolidine and the weight ratio of 2,2-dibromomalonamide to 7-ethyl bicyclooxazolidine is from about 5:1 to about 1:1500, alternatively from about 1:1 to about 1:1300, or alternatively from about 1:1 to about 1:1280, or alternatively from about 1:1.2 to about 1:1280.
In some embodiments, the aldehyde-based biocidal compound is 1-(3-chloroallyl)-3,5,7-triaza-1-azoniaadamantane chloride and the weight ratio of 2,2-dibromomalonamide to 1-(3-chloroallyl)-3,5,7-triaza-1-azoniaadamantane chloride is from about 10:1 to about 1:350, alternatively from about 4:1 to about 1:320, or alternatively from about 3.2:1 to about 1:320.
In some embodiments, the aldehyde-based biocidal compound is 4,4-dimethyloxazolidine and the weight ratio of 2,2-dibromomalonamide to 4,4-dimethyloxazolidine is from about 5:1 to about 1:5, alternatively from about 1:2 to about 1:3, or alternatively about 1:2.5.
In some embodiments, the aldehyde-based biocidal compound is 1,3,5-triethylhexahydro-s-triazine and the weight ratio of 2,2-dibromomalonamide to 1,3,5-triethylhexahydro-s-triazine is from about 10:1 to about 1:30, alternatively from about 4:1 to about 1:20, or alternatively from about 2:1 to about 1:16.
The composition of the invention is useful for controlling microorganisms in a variety of aqueous and water containing systems. Examples of such systems include, but are not limited to, paints and coatings, aqueous emulsions, latexes, adhesives, inks, pigment dispersions, household and industrial cleaners, detergents, dish detergents, mineral slurries polymer emulsions, caulks and adhesives, tape joint compounds, disinfectants, sanitizers, metalworking fluids, construction products, personal care products, textile fluids such as spin finishes, industrial process water (e.g. oilfield water, pulp and paper water, cooling water), oilfield functional fluids such as drilling muds and fracturing fluids, fuels, air washers, wastewater, ballast water, filtration systems, and swimming pool and spa water. Preferred aqueous systems are metal working fluids, personal care, household and industrial cleaners, industrial process water, and paints and coatings. Particularly preferred are industrial process water, paints and coatings, metal working fluids, and textile fluids such as spin finishes.
A person of ordinary skill in the art can readily determine, without undue experimentation, the effective amount of the composition that should be used in any particular application to provide microorganism control. By way of illustration, a suitable actives concentration (total for both 2,2-dibromomalonamide and aldehyde-based biocidal compound) is typically at least about 1 ppm, alternatively at least about 3 ppm, alternatively at least about 7 ppm, alternatively at least about 10 ppm, or alternatively at least about 100 ppm based on the total weight of the aqueous or water containing system. In some embodiments, a suitable upper limit for the actives concentration is about 1000 ppm, alternatively about 500 ppm, alternatively about 100 ppm, alternatively about 50 ppm, alternatively about 30 ppm, alternatively about 15 ppm, alternatively about 10 ppm, or alternatively about 7 ppm, based on the total weight of the aqueous or water containing system.
The components of the composition can be added to the aqueous or water containing system separately, or pre-blended prior to addition. A person of ordinary skill in the art can easily determine the appropriate method of addition. The composition can be used in the system with other additives such as, but not limited to, surfactants, ionic/nonionic polymers and scale and corrosion inhibitors, oxygen scavengers, and/or additional biocides.
The following examples are illustrative of the invention but are not intended to limit its scope. Unless otherwise indicated, the ratios, percentages, parts, and the like used herein are by weight.
EXAMPLES
The results provided in the Examples are generated using a growth inhibition assay or a kill assay. Details of each assay are provided below.
Kill Assay.
This assay is used as a preliminary evaluation of synergy between the actives. The procedure is as follows. A mineral salts solution (0.2203 g of CaCl2, 0.1847 g of MgSO4, and 0.2033 g of NaHCO3 in 1 L water, approximately pH 8) is inoculated with equal amounts (about 107 CFU/ml) of a mixture of Pseudomonas aeruginosa ATCC 10145 and Staphylococcus aureus ATCC 6538. Aliquots of the cell suspension are then treated with 2,2-dibromomalonamide (“DBMAL”), an aldehyde-based biocidal compound, and their combinations at various concentration levels. After incubating at 37° C. for 2 hours, the biocidal efficacy is determined on the basis of the minimum biocide concentration (MBC) needed to completely kill the bacterial cells in the aliquots. The MBC values are then used to calculate a synergy index (SI) values.
Summaries of the kill assay results are presented in the individual Examples. In each table, MBC values for each biocide and the blends tested are presented. Likewise, the Synergy Index (“SI”) values for the combinations are listed. SI is calculated with the following equation:

Synergy Index=Ca/CA+Mb/CB 

where

    
    
        Ca: Concentration of biocide A required for complete bacterial kill when used in combination with biocide B
        CA: Concentration of biocide A required for complete bacterial kill when used alone
        Cb: Concentration of biocide B required for complete bacterial kill when used in combination with biocide A
        CB: Concentration of biocide B required for complete bacterial kill when used alone

The SI values are interpreted as follows:

        SI<1: Synergistic
        SI=1: Additive
        SI>1: Antagonistic
    
    


Growth Inhibition Assay.
The growth inhibition assay used in the Examples measures inhibition of growth (or lack thereof) of a microbial consortium. Inhibition of growth can be the result of killing of the cells (so no growth occurs), killing of a significant portion of the populations of cells so that regrowth requires a prolonged time, or inhibition of growth without killing (stasis). Regardless of the mechanism of action, the impact of a biocide (or combination of biocides) can be measured over time on the basis of an increase in the size of the community.
The assay measures the efficacy of one or more biocides at preventing growth of a consortium of bacteria in a dilute mineral salts medium. The medium contains (in mg/l) the following components: FeCl3.6H2O (1); CaCl2.2H2O (10); MgSO4.7H2O (22.5); (NH4)2SO4 (40); KH2PO4 (10); K2HPO4 (25.5); Yeast Extract (10); and glucose (100). After all components are added to deionized water, the pH of the medium is adjusted to 7.5. Following filter sterilization, aliquots are dispensed in 100 ul quantities to sterile microtiter plate wells. Dilutions of DBMAL and/or “Biocide B” are then added to the microtiter plate. After preparing the combinations of actives as illustrated below, each well is inoculated with 100 μl of a cell suspension containing ca. 1×106 cells per milliliter of a mixture of Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, and Bacillus subtilis. The final total volume of medium in each well is 300 μl. Once prepared as described herein, the concentration of each active ranges from 25 ppm to 0.19 ppm as illustrated in Table 1. The resulting matrix allows testing of eight concentrations of each active and 64 combinations of actives in the ratios (of actives).







TABLE 1







Template for microtiter plate-based synergy assay showing concentrations 


of each active. Ratios are based on weight (ppm) of each active.











Biocide B (mg/l)


















25.000
12.500
6.250
3.13
1.56
0.78
0.39
0.19





DBMAL
25.000
 1:1
 1:2
 1:4
 1:8
1:16
1:32
1:64
1:128


(mg/l)
12.500
 2:1
 1
 1:2
 1:4
1:8
1:16
1:32
1:64



6.250
 4:1
 2:1
 1
 1:2
1:4
1:8
1:16
1:32



3.13
 8:1
 4:1
 2:1
 1
1:2
1:4
1:8
1:16



1.56
 16:1
 8:1
 4:1
 2:1
1
1:2
1:4
1:8



0.78
 32:1
16:1
 8:1
 4:1
2:1
1
1:2
1:4



0.39
 64:1
32:1
16:1
 8:1
4:1
2:1
1
1:2



0.19
128:1
64:1
32:1
16:1
8:1
4:1
2:1
1:1









Controls (not shown) contain the medium with no biocide added. After preparing the combinations of actives as illustrated above, each well is inoculated with 100 μl of a cell suspension containing ca. 1×106 cells per milliliter of a mixture of Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, and Bacillus subtilis. The final total volume of medium in each well is 300 μl.

Immediately after the microtiter plates are prepared, the optical density (OD) readings for each well is measured at 580 nm and the plates are then incubated at 37° C. for 24 hr. After the incubation period, the plates are gently agitated before OD580 values are collected. The OD580 values at To are subtracted from T24 values to determine the total amount of growth (or lack thereof) that occurs. These values are used to calculate the percent inhibition of growth caused by the presence of each biocide and each of the 64 combinations. A 90% inhibition of growth is used as a threshold for calculating synergy index (SI) values with the following equation:

Synergy Index=MDBMAL/CDBMAL+MB/CB 

where

    
    
        CDBMAL: Concentration of DBMAL required to inhibit  ̃90% of bacterial growth when used alone
        CB: Concentration of biocide (B) required to inhibit  ̃90% of bacterial growth when used alone.
        MDBMAL: Concentration of DBMAL required to inhibit  ̃90% of bacterial growth when used in combination with biocide (B).
        MB: Concentration of biocide (B) required to inhibit  ̃90% of bacterial growth when used in combination with DBMAL

The SI values are interpreted as follows:

        SI<1: Synergistic combination
        SI=1: Additive combination
        SI>1: Antagonistic combination
    
    


In the Examples below, the amounts of biocides in the solution are measured in mg per liter of solution (mg/l). Since solution densities are approximately 1.00, the mg/l measurement corresponds to weight ppm. Both units may therefore be used interchangeably in the Examples.
Example 1
DBMAL and Glutaraldehyde
Kill Assay Results. The kill assay result on combinations of DBMAL and glutaraldehyde (Glut) are presented in Table 2. In this assay, when tested alone, the concentrations of DBMAL and glutaraldehyde needed to achieve complete kill of the test strains is 66.7 mg/l and 29.6 mg/l, respectively. However, when tested together in a 1:1 blend, only 14.8 mg/l of each active is required; the corresponding SI is 0.72. Other combinations of DBMAL and glut exhibit synergy.







TABLE 2







MBC of DBMAL, Glut, and combinations thereof













1st
2nd





biocide
biocide




Active
DBMAL
Glut




weight ratio 
Concn.
Concn.
Synergy



(1st:2nd)
(mg/l)
(mg/l)
Index
















DBMAL
66.7
0.0




alone






9:1
40.0
4.4
0.75



3:1
14.8
4.9
0.57



1:1
14.8
14.8
0.72



1:3
7.4
22.2
0.86



1:9
3.0
26.7
0.95



Glut alone
0.0
29.6










Growth Inhibition Assay Results. Table 3 shows the assay results for DBMAL, glutaraldehyde, and combinations. The concentration of glutaraldehyde needed to provide at least 90% inhibition of growth of the microorganism consortium is 125 mg/l and that for DBMAL is 12.5 mg/l.







TABLE 3







Percent inhibition of growth in a species-defined microbial consortium by glutaraldehyde (glut) and DBMAL


alone and combinations of these actives after a 24-hour incubation period. Numbers represent percent inhibition


of growth as measured by optical density measurements (580 nm) at time = 24 hours compared with time = 0 values.










Single Actives (mg/l)











%

%













Inhibition

Inhibition

%



of growth

of

Inhibition
Combinations of DBMAL and Glut













in
DBMAL
growth
Glut
of
DBMAL



Untreated
Concn.
by
Concn.
growth
Concn.
Glut Concn. (mg/l)




















Control
(mg/l)
DBMAL
(mg/l)
by Glut.
(mg/l)
500.0
250.0
125.0
62.5
31.25
15.63
7.81
3.91























22
25.0
105
500.0
100
25.0
97
98
93
100
99
100
100
100


14
12.5
107
250.0
100
12.5
98
100
100
100
100
100
100
100


0
6.25
50
125.0
100
6.25
100
100
93
100
100
50
23
16


0
3.13
38
62.5
0
3.13
64
100
100
100
91
43
6
0


0
1.56
11
31.25
21
1.56
99
100
100
100
27
26
5
0


8
0.78
34
15.63
20
0.78
99
99
97
100
74
18
2
0


0
0.39
0
7.81
18
0.39
97
100
100
100
68
13
0
0


22
0.19
0
3.91
0
0.19
95
100
100
100
70
12
0
0









Table 4 shows ratios of DBMAL and glutaraldehyde found to be synergistic under the growth inhibition assay.










TABLE 4





DBMAL
Glut Concn.
Ratio
Synergy Index


Concn. (mg/l)
(mg/l)
(DBMAL to Glut)
(SI)


















6.25
31.25
1:5 
0.75


3.13
62.5
1:20 
0.75


3.13
31.25
1:10 
0.50


1.56
62.5
1:40 
0.63


1.56
31.25
1:20 
0.37


0.78
62.5
1:80 
0.56


0.39
62.5
1:160
0.53


0.19
62.5
1:320
0.52









Example 2
DBMAL and CTAC
Inhibition Growth Assay Results. Table 5 shows the inhibition growth assay results for DBMAL, 1-(3-chloroallyl)-3,5,7-triaza-1-azoniaadamantane chloride (“CTAC”), and combinations.







TABLE 6







Percent inhibition of growth in a species-defined microbial consortium by CTAC and DBMAL alone


and combinations of these actives after a 24-hour incubation period. Numbers represent percent inhibition 


of growth as measured by optical density measurements (580 nm) at time = 24 hours compared with time = 0 values.


















Single Actives (mg/l)





























%

%

%











Inhibition

Inhibition

Inhibition





















of growth

of 

of
Combinations of DBMAL and CTAC




















in
DBMAL
growth
CTAC
growth
DBMAL





















Untreated 
Concn.
by
Concn.
by
Concn.
CTAC Concn. (mg/l)




















Control
(mg/l)
DBMAL
(mg/l)
CTAC
(mg/l)
250.0
125.0
62.5
31.25
15.625
7.8125
3.91
1.95























20
25
100
250.0
98
25.0
100
99
97
99
99
100
99
99


5
12.5
97
125.0
100
12.5
100
100
100
99
100
100
99
100


0
6.25
44
62.5
41
6.25
100
100
100
100
100
100
100
100


0
3.13
50
31.25
35
3.13
100
100
100
100
100
86
40
22


12
1.56
34
15.625
0
1.56
99
99
100
100
31
18
6
0


20
0.78
18
7.81
9
0.78
100
100
99
89
3
3
0
0


3
0.39
0
3.91
0
0.39
100
100
99
39
0
0
0
0


0
0.19
0
1.95
0
0.19
99
100
98
0
0
0
0
0









Table 7 shows ratios of DBMAL and CTAC found to be synergistic under the growth inhibition assay. Ratios are based on concentrations of the actives.










TABLE 7





DBMAL Concn.
CTAC Concn.
Ratio
Synergy Index


(mg/l)
(mg/l)
(DBMAL:CTAC)
(SI)


















6.25
31.25
1:5  
0.75


6.25
15.63
1:2.5
0.63


6.25
7.81
 1:1.25
0.56


6.25
3.91
1.6:1
0.53


6.25
1.95
3.2:1
0.52


3.13
62.50
1:20 
0.75


3.13
31.25
1:10 
0.50


3.13
15.63
1:5 
0.38


1.56
62.50
1:40 
0.62


1.56
31.25
1:20 
0.37


0.78
62.50
1:80 
0.56


0.39
62.50
1:160
0.53


0.19
62.50
1:320
0.52









Example 3
DBMAL and TN
Inhibition Growth Assay Results. Table 8 shows the inhibition growth assay results for DBMAL, tris(hydroxymethyl)nitromethane (“TN”), and combinations thereof.







TABLE 8







Percent inhibition of growth in a species-defined microbial consortium by TN and DBMAL


alone and combinations of these actives after a 24-hour incubation period. Numbers represent percent 


inhibition of growth as measured by optical density measurements (580 nm) at time = 24 hours compared with time = 0 values.

















Single Actives (mg/l)




























%

%













Inhibition 

Inhibition

%





















of growth

of

Inhibition
Combinations of DBMAL and TN




















in
DBMAL
growth
TN
of
DBMAL





















Untreated 
Concn.
by
concn.
growth 
concn.
TN concn. (mg/l)




















Control
(mg/l)
DBMAL
(mg/l)
by TN
(mg/l)
1000.0
500.0
250.0
125.0
62.5
31.25
15.63
7.81























0
25.0
100
1000.0
100
25.0
100
100
100
100
100
100
100
100


2
12.5
100
500.0
100
12.5
100
100
100
100
100
100
100
100


2
6.25
0
250.0
100
6.25
100
100
100
100
100
100
100
0


2
3.13
0
125.0
100
3.13
100
100
100
100
100
100
0
0


5
1.56
0
62.5
50
1.56
100
100
100
100
100
0
0
0


2
0.78
0
31.25
0
0.78
100
100
100
100
76
0
0
0


0
0.39
2
15.63
4
0.39
100
100
100
100
0
0
0
0


0
0.19
0
7.81
0
0.19
100
100
100
100
0
0
0
0









Table 9 shows concentrations of concentrations of DBMAL and TN found to be synergistic. The ratios are based on concentrations (in mg/l) of the actives.










TABLE 9





DBMAL concn.
TN concn.
Ratio
Synergy Index


(mg/l)
(mg/l)
(DBMAL to TN)
(SI)


















6.25
31.25
1:5 
0.75


6.25
15.63
 1:2.5
0.63


3.13
62.5
1:20
0.75


3.13
31.25
1:10
0.5


1.56
62.5
1:40
0.63









Example 4
DBMAL and EBO
Inhibition Growth Assay Results. Table 10 shows the inhibition growth assay results for DBMAL, 7-ethyl bicyclooxazolidine (“EBO”), and combinations thereof.







TABLE 10







Percent inhibition of growth in a species-defined microbial consortium by 7-ethyl bicyclooxazolidine 


(“EBO”) and DBMAL alone and combinations of these actives after a 24-hour incubation period. Numbers represent 


percent inhibition of growth as measured by optical density measurements (580 nm) at time = 24 hours compared with time = 0 values.










Single Actives (mg/l)













%

%





Inhibition

Inhibition

%



of growth

of

Inhibition
Combinations of DBMAL and EBO













in
DBMAL
growth
EBO
of
DBMAL



Untreated
concn.
by
concn.
growth
concn.
EBO concn. (mg/l)




















Control
(mg/l)
DBMAL
mg/l
by EBO
(mg/l)
1000.0
500.0
250.0
125.0
62.5
31.25
15.63
7.81























8
25.0
100
1000.0
100
25
98
99
98
100
100
99
100
99


10
12.5
99
500.0
96
12.5
99
99
97
100
87
99
91
99


5
6.25
52
250.0
71
6.25
100
99
97
99
99
94
98
95


0
3.13
0
125.0
10
3.13
98
83
97
93
0
0
0
0


0
1.56
0
62.5
3
1.56
98
98
99
31
0
0
0
0


0
0.78
0
31.25
0
0.78
98
99
97
0
0
0
0
0


6
0.39
0
15.63
0
0.39
96
99
98
0
0
0
0
0


2
0.19
7
7.81
0
0.19
90
100
97
0
0
0
0
0









Table 11 shows concentrations of Concentrations of DBMAL and EBO found to be synergistic. The ratios are based on concentrations (in mg/l) of the actives.












TABLE 11









Ratio




DBMAL concn.
EBO concn.
(DBMAL to
Synergy Index



(mg/l)
(mg/l)
EBO)
(SI)





















6.25
125
1:20
0.75



6.25
62.5
1:10
0.63



6.25
31.25
1:5 
0.56



6.25
15.63
 1:2.5
0.53



6.25
7.81
 1:1.2
0.52



3.13
250
1:80
0.75



1.56
125
1:80
0.38



0.78
250
 1:320
0.56



0.39
250
 1:640
0.54



0.19
250
  1:1280
0.52










Example 5
DBMAL and 4,4-Dimethyloxazolidine
Growth Inhibition Assay Results. Table 12 shows the inhibition growth assay results for DBMAL, 4,4-Dimethyloxazolidine (“4,4-D”), and combinations thereof.







TABLE 12







Percent inhibition of growth in a species-defined microbial consortium by 4,4-Dimethyloxazolidine (“4,4-D”) 


and DBMAL alone and combinations of these actives after a 24-hour incubation period. Numbers represent percent 


inhibition of growth as measured by optical density measurements (580 nm) at time = 24 hours compared with time = 0 values.

















%
Single Actives (mg/l)





























Inhibition

%























of 

Inhibition

%
Combinations of DBMAL and 4,4-D




















growth in 
DBMAL
of growth 
4,4-D
Inhibition
DBMAL





















Untreated 
Concn.
by
Concn.
of growth
Concn.
4,4-D Concn. (mg/l)




















Control
(mg/l)
DBMAL
(mg/l)
by 4,4-D
(mg/l)
1000.0
500.0
250.0
125.0
62.5
31.25
15.63
7.81























2
25.0
100
1000.0
100
25.0
99
100
99
100
100
100
100
100


1
12.5
100
500.0
100
12.5
100
100
100
100
100
100
100
100


0
6.25
0
250.0
99
6.25
100
100
100
100
100
100
100
0


0
3.13
0
125.0
100
3.13
100
100
100
100
100
51
0
0


5
1.56
2
62.5
98
1.56
100
100
100
100
100
0
0
0


0
0.78
0
31.25
13
0.78
100
100
2
100
100
0
0
0


1
0.39
6
15.63
2
0.39
100
100
100
100
85
0
0
0


0
0.19
0
7.81
0
0.19
100
100
0
100
100
0
0
0









Table 13 shows concentrations of Concentrations of DBMAL and 4,4-Dimethyloxazolidine (“4,4-D”) found to be synergistic. The ratios are based on concentrations (in mg/l) of the two actives.












TABLE 13







DBMAL 
4,4-D Concn.
Ratio (DBMAL 
Synergy 



Concn. (mg/l)
(mg/l)
to 4,4-D)
Index (SI)









6.25
15.63
1:2.5
0.75










Example 6
DBMAL and 1,3,5-Triethylhexahydro-s-triazine
Growth Inhibition Assay Results. Table 12 shows the inhibition growth assay results for DBMAL, 1,3,5-Triethylhexahydro-s-triazine (“TEHT”), and combinations thereof.







TABLE 12







Percent inhibition of growth in a species-defined microbial consortium by 


1,3,5-Triethylhexahydro-s-triazine (“TEHT”) and DBMAL alone and combinations of 


these actives after a 24-hour incubation period. Numbers represent percent inhibition of growth 


as measured by optical density measurements (580 nm) at time = 24 hours compared with time = 0 values.

















%
Single Actives (mg/l)





























Inhibition

%























of growth

Inhibition

%
Combinations of DBMAL and TEHT




















in
DBMAL
of growth
TEHT
Inhibition
DBMAL





















Untreated
Concn.
by
Concn.
of growth
Concn.
TEHT concn. (mg/l)




















Control
(mg/l)
DBMAL
(mg/l)
by TEHT
(mg/l)
25
12.5
6.25
3.13
1.56
0.78
0.39
0.19























1
25.0
96
25.0
100
25.0
100
100
100
100
100
100
100
100


0
12.5
97
12.5
98
12.5
100
100
100
99
100
99
99
99


0
6.25
32
6.25
82
6.25
100
100
100
100
50
12
5
12


0
3.13
45
3.13
49
3.13
100
100
100
75
0
0
0
0


2
1.56
32
1.56
22
1.56
100
100
92
18
0
13
8
0


0
0.78
30
0.78
10
0.78
100
100
100
20
0
9
0
0


0
0.39
8
0.39
11
0.39
99
91
100
0
0
0
0
0


3
0.19
13
0.19
10
0.19
100
91
56
0
0
0
0
0









Table 13 shows concentrations of Concentrations of DBMAL and 1,3,5-Triethylhexahydro-s-triazine (“TEHT”) found to be synergistic. The ratios are based on concentrations (in mg/l) of the two actives.












TABLE 13







DBMAL concn.
TEHT concn.





(mg/l)
(mg/l)
SI
Ratio





















3.13
6.25
0.75
1:2



1.56
6.25
0.63
1:4



0.78
6.25
0.56
1:8



0.39
6.25
0.53
 1:16



6.250
3.13
0.75
2:1










While the invention has been described above according to its preferred embodiments, it can be modified within the spirit and scope of this disclosure. This application is therefore intended to cover any variations, uses, or adaptations of the invention using the general principles disclosed herein. Further, the application is intended to cover such departures from the present disclosure as come within the known or customary practice in the art to which this invention pertains and which fall within the limits of the following claims.

## Claims

1. A biocidal composition for controlling microorganisms in an aqueous or water-containing system, the composition comprising: 2,2-dibromomalonamide and an aldehyde-based biocidal compound;
wherein the aldehyde-based biocidal compound is 1-(3-chloroallyl)-3,5,7-triaza-1-azoniaadamantane chloride and the weight ratio of 2,2-dibromomalonamide to 1-(3-chloroallyl)-3,5,7-triaza-1-azoniaadamantane chloride is from about 4:1 to about 1:320.

2. A composition according to claim 1 which is: paint, coating, aqueous emulsion, latex, adhesive, ink, pigment dispersion, household or industrial cleaner, detergent, dish detergent, mineral slurry polymer emulsion, caulk, adhesive, tape joint compound, disinfectant, sanitizer, metalworking fluid, construction product, personal care product, textile fluid, spin finish, industrial process water, oilfield functional fluid, fuel, air washer, wastewater, ballast water, filtration systems, or swimming pool and spa water.

3. A method for controlling microorganism growth in an aqueous or water-containing system, the method comprising treating the aqueous or water-containing system with an effective amount of a composition according to claim 1.

4. A method according to claim 3 wherein the aqueous or water-containing system is paint, coating, aqueous emulsion, latex, adhesive, ink, pigment dispersion, household or industrial cleaner, detergent, dish detergent, mineral slurry polymer emulsion, caulk, adhesive, tape joint compound, disinfectant, sanitizer, metalworking fluid, construction product, personal care product, textile fluid, spin finish, industrial process water, oilfield functional fluid, fuel, air washer, wastewater, ballast water, filtration system, or swimming pool and spa water.

5. A method according to claim 3 wherein the composition inhibits the growth of microorganisms in the aqueous or water-containing system.

6. A method according to claim 3 wherein the composition kills microorganisms in the aqueous or water-containing system.