Synergistic antimicrobial compositions containing 1-hydroxymethylpyrazole and a preservative

Abstract

Compositions comprising a compound of formula I ##STR1## where R and R' are independently selected from hydrogen and alkyl of 1 to 4 carbon atoms, R" is a moiety selected from the group consisting of (a) hydrogen, (b) a halogen atom selected from the group consisting of F, Cl, Br, and I, and (c) a nitro group, or the hydrochloride salt of said compound; and at least one preservative are disclosed which are synergistically effective compared to the respective components alone in controlling and/or reducing the growth of microorganisms in or on a material or medium. Methods to control and/or reduce the growth of microorganisms and prevent spoilage caused by microorganisms with the use of the compositions of the present invention are also disclosed.

Description

The present invention relates to certain compositions and processes useful for controlling the growth of one or more microorganisms and for preventing spoilage caused by bacteria and/or fungi in various products, materials, or media, particularly industrial products, materials or media. These materials or media include for example, wood pulp, wood chips, lumber, adhesives, coatings, animal hides including birds and reptiles, paper mill liquors, paper, optical brighteners, pharmaceutical and chemical formulations, cosmetics and toiletry formulations, geological drilling lubricants and aids, agrochemical compositions, paints, leathers, wood, metalworking fluids, industrial process aid formulations, cooling tower water, waste water, pasteurizers, tanning liquors, starch, dyes, petrochemicals, surfactants, defoamers, emulsions or water containing polymers, lipid based materials, carbohydrate based materials, proteinaceous materials, acrylic latex paint emulsions, and textiles. The novel processes and mixtures of the present invention show unexpected synergistic activity against microorganisms, including bacteria and fungi. Specifically, the invention is directed to the use of compositions comprising dimethylhydroxymethylpyrazole and at least one preservative.

Many of the products, materials, or media referred to above, when wet, subjected to treatment in water, or placed in a humid environment, are susceptible to bacterial and/or fungal deterioration or degradation unless steps are taken to control such degradation or deterioration.

To control deterioration or degradation caused by microorganisms, various microbicides are used. Workers in the trade have continued to seek improved biocides that have low toxicity and are capable of exhibiting a prolonged biocidal effect at normal use levels.

Preservatives can be used alone to control microorganisms, but many of them have low efficacy against bacteria and fungi unless extremely high concentrations are used. 1-hydroxymethylpyrazoles can also be used alone in low concentrations as a biocide.

This invention provides a microbicidal composition capable of controlling the growth of at least one microorganism, particularly fungi and bacteria, over prolonged periods of time. Additionally, the present invention provides compositions which are economical to use. Methods of controlling the growth of at least one microorganism are also objects of this invention.

The present invention provides a composition comprising a 1-hydroxymethylpyrazole and at least one preservative, where the components are present in a combined amount synergistically effective to control the growth of at least one microorganism.

The present invention also provides a method for controlling the growth of at least one microorganism in or on a material or medium susceptible to attack by the microorganism. That method comprises the step of adding to the material or medium a composition of the present invention in an amount synergistically effective to control the growth of the microorganism.

The present invention also embodies the separate addition of a 1-hydroxymethylpyrazole and at least one preservative to the products, materials or media described above. According to this embodiment, the components are individually added to the system so that the final amount of the 1-hydroxymethylpyrazole and at least one preservative present in the system at the time of use is that amount synergistically effective to control the growth of at least one microorganism.

The compositions of the present invention are also useful in preserving various types of industrial media or materials susceptible to attack by microorganisms. Such media or materials include, but are not limited to, wood pulp, wood chips, lumber, adhesives, coatings, animal hides including birds and reptiles, paper mill liquors, paper, optical brighteners, pharmaceutical and chemical formulations, cosmetics and toiletry formulations, geological drilling lubricants and aids, agrochemical compositions, paints, leathers, wood, metalworking fluids, industrial process aid formulations, cooling tower water, waste water, pasteurizers, tanning liquors, starch, dyes, petrochemicals, surfactants, defoamers, emulsions or water containing polymers, lipid based materials, carbohydrate based materials, proteinaceous materials, acrylic latex paint emulsions, and textiles.

The composition can also be useful in agrochemical formulations for the purpose of protecting seeds or crops against microbial spoilage.

Additional objects and advantages of the present invention will be set forth in part in the description which follows, and in part will be apparent from the description, or may be learned by the practice of the present invention. The objects and advantages of the present invention may be realized and obtained by means of the elements and combinations particularly pointed out in the claims.

It is to be understood that both the foregoing general description and the following general description are exemplary and explanatory only and are not restrictive of the present invention as claimed.

When two chemical microbicides are combined into one product or added separately three results are possible:

1) The resulting product would produce an additive (neutral) effect. 2) The products in the product would produce an antagonistic effect, or 3) The chemicals in the product would produce a synergistic effect.

An additive effect has no economic advantage over the individual components. The antagonistic effect would produce a negative impact. Only a synergistic effect, which is less likely than either an additive or antagonistic effect, would produce a positive effect and therefore possess economic advantages.

The microbicidal compositions combining a 1-hydroxymethylpyrazole and at least one preservative demonstrate an unexpected synergistic effect compared to the respective components alone. Thus, these compositions achieve superior, i.e. greater than additive, microbicidal activity at low concentrations against a wide variety of microorganisms. Examples of microorganisms include, but are not limited to, fungi and bacteria such as Aspergillus niger (ATCC 9642) and Pseudomonas aeruginosa (ATCC 15442). Preferably, the compositions of the present invention have a low toxicity.

The 1-hydroxymethylpyrazole is a compound having the formula ##STR2## wherein R and R' are independently selected from hydrogen and alkyl of 1 to 4 carbon atoms, R" is a moiety selected from the group consisting of (a) hydrogen, (b) a halogen atom selected from the group consisting of F, Cl, Br, and I, and (c) a nitro group, or the hydrochloride salt of said compound.

The preparation of 1-hydroxymethylpyrazole is described in U.S. Pat. Nos. 4,801,361 and 5,335,432, and references referred to in these patents, all of which are incorporated herein by reference. 3,5-dimethyl-1-hydroxymethylpyrazole is commercially available and sold under the tradename Busan.RTM. 1504 or Busan.RTM. 1104 (Buckman Laboratories International, Memphis, Tenn.) and also easily synthesized from commercially available raw materials.

As described in Buckman Laboratories' Product Profile and Analytical Method (1992) for Busan.RTM. 1504 (both incorporated herein in their entirety by reference), Busan.RTM. 1504 is a broad-spectrum, nonionic bactericide and fungicide developed for the preservation of aqueous and aqueous-emulsified systems. Busan.RTM. 1504 is active over a broad pH range and is stable under both alkaline and acidic conditions. In addition, Busan.RTM. 1504 can be formulated into cationic, anionic, and nonionic systems. Busan.RTM. 1504 is also compatible with proteins. Busan.RTM. 1504 is a water-soluble, crystalline solid having 1-hydroxymethyl-3,5-dimethyl pyrazole as an active ingredient.

The preservative can be any preservative that will produce a synergistic effect when combined with a 1-hydroxymethylpyrazole. Examples include hexahydro-1,3,5-tris(2-hydroxyethyl)-s-triazine, bis 1,4-(bromoacetoxy)-2-butene; 2-bromo-2-nitropropane-1,3 diol; dimethylhydantoin; imidazolididinyl urea (also known as N,N.sup.1 -methylene bis(N.sup.1 -[1-hydroxy methyl)-2,5-dioxo-4-imidazolidinyl] urea); diazolidinyl urea (also known as N-(hydroxymethyl)-N-(1,3-dihydroxymethyl-2-5-dioxo-4-imidozolidinyl)-N-(hydroxymethyl) urea); mixture of 5-chloro-2-methyl-4-iso-thiazoline-3-one and 2-methyl-4-iso-thiazoline-3-one; 1-(3-chloroallyl)-3,5,7-triaza-1-azoniaadamantane chloride; and C.sub.1 -C.sub.6 alkyl hydroxybenzoate. Further, mixtures of these preservatives can also be used. When such mixtures are used in combination with dimethylhydroxymethylpyrazole, at least one of the preservatives in the mixtures has a synergistic relationship with the dimethylhydroxymethylpyrazole. Preservatives useful in the invention are commercially available or may be synthesized from commercially available raw materials. Examples of commercially available preservatives include, but are not limited to, Busan.RTM. 1210 (bis 1,4-(bromoacetoxy)-2-butene--available from Buckman Laboratories international), Kathon CG (mixture of 5-chloro-2-methyl-4-isothiazoline-3-one and 2-methyl-4-isothiazoline-3-one--available from Rohm & Haas), Dowicil 200 (CIS isomer of 1-(3-chloroallyl)-3,5,7-triaza-1-azonia adamantane chloride--available from Dow Chemical Co.), Propyl Paraben (C.sub.1 -C.sub.6 alkyl hydroxybenzoate--available from various sources), Busan.RTM. 1506 (hexahydro-1,3,5-tris(2-hydroxy-ethyl)-5-triazine--available from Buckman Laboratories International), Sodium Omadine (sodium 2-pyridinethiol-1-oxide--available from Olin Chemical Co.), Germall II (N-(hydroxymethyl)-N-C1,3-dihydroxymethyl-2-5-dioxo-4-imidozolidinyl)-N-(hydroxymethyl) urea or diazolidinyl urea--available from Sutton Laboratories), Bronopol (2-bromo-2-nitropropane-1,3 diol--available from Boots Chemical, and Glydant (dimethyl hydantoin--available from Sutton Laboratories).

The preservative may be chosen based on the compatibility with the materials or media. Compatibility is determined by criteria such as solubility in the fluid system and lack of reactivity with the fluid, material, or media in question. In view of the present invention, the compatibility is readily determined by one having ordinary skill in the art by adding (e.g., applying on the surface, incorporating in the fluid, material or media, and the like) the preservative to the material or media to be used. When used in a fluid system it is preferable that the preservative be freely soluble in the particular fluid resulting in a uniform solution or dispersion.

As these terms are used herein, "preventing" (which includes mitigating) spoilage is to be understood that the present invention in effect "controls" the growth of at least one microorganism, responsible, at least in part, for the spoilage. It is to be further understood that by "controlling" (i.e., preventing), the growth of at least one of these types of microorganisms is inhibited. In other words, there is no growth or essentially no growth of at least one microorganism. "Controlling" the growth of at least one microorganism maintains the microorganism population at a desired level, reduces the population to a desired level (even to undetectable limits, e.g., zero population), and/or inhibits the growth of the microorganism. Thus, the substrates or materials susceptible to attack by these types of microorganisms are preserved from this attack and the resulting spoilage and other detrimental effects caused by the microorganisms. Further, it is also to be understood that "controlling" the growth of at least one microorganism also includes biostatically reducing and/or maintaining a low level of microorganisms such that the attack by microorganisms and any resulting spoilage or other detrimental effects are mitigated, i.e., the microorganism growth rate or microorganism attack rate is slowed down or eliminated.

In the following discussion of preferred embodiments, component (a) is 3,5,-dimethyl 1-hydroxymethylpyrazole and component (b) is a preservative.

As described above, components (a) and (b) are used in synergistically effective amounts. The weight ratios of (a) to (b) vary depending on the type of microorganisms, material, or media to which the composition is applied. Generally, the ratio of component (a) to component (b) preferably ranges from 0.1 to 99, more preferably from 1:30 to 30:1, and most preferably from 1:5 to 5:1.

The following weight ratios of dimethylhydroxymethylpyrazole to the following preservatives are also preferred:

______________________________________Busan .RTM. 1504: Busan .RTM. 1210 80:0.01 to 1:2Busan .RTM. 1504: Kathon CG .RTM. 80:1 to 1:2Busan .RTM. 1504: Dowicil 200 .TM. 10:1 to 1:20Busan .RTM. 1504: Propyl Paraben 8:1 to 1:10Busan .RTM. 1504: Methyl Paraben 8:1 to 1:80Busan .RTM. 1504: Busan .RTM. 1506 16:1 to 1:30Busan .RTM. 1504: Sodium Omadine 50:0.1 to 1:10Busan .RTM. 1504: Germall II .RTM. 4:1 to 0.1:25Busan .RTM. 1504: Germall 115 .RTM. 0.8:1 to 0.1:16Busan .RTM. 1504: Bronopol .RTM. 80:1 to 1:60Busan .RTM. 1504: Glydant .RTM. 7:1 to 0.1:25______________________________________

Depending upon the specific application, the composition may be prepared in liquid form by dissolving the composition in an organic solvent. The preservative may be prepared in an emulsion form by emulsifying it in water, or if necessary, by adding a surfactant. Additional chemicals such as corrosion inhibitors, performing agents, and insecticides may be added to the foregoing preparation.

The synergistic activity of the combinations described above has been confirmed using standard laboratory techniques as illustrated below. The following examples are intended to illustrate, not limit, the present invention.

MICROBIOLOGICAL EVALUATION

A. Fungal Evaluation

Mineral salts-glucose medium was used. To prepare the medium, the following ingredients were added to 1 liter of deionized water: 0.7 g of KH.sub.2 PO.sub.4, 0.7 g of MgSO.sub.4.7H.sub.2 O, 1.0 g of NH.sub.4 NO.sub.3, 0.005 g NaCl, 0.002 g FeSO.sub.4.7H.sub.2 O, 0.002 g ZnSO.sub.4.7H.sub.2 O, 0.001 g MnSO.sub.4.7H.sub.2 O, 10 g of Glucose. The pH of the medium was adjusted to 6 with 1N NaOH. The medium was distributed in 5.0 ml amounts in test tubes and autoclaved at 121.degree. C. for 20 minutes. The fungus, A. niger, was grown on a potato dextrose agar slant for 7 to 10 days and a spore suspension prepared by washing down the spores from the slant into a sterile saline solution. After addition of the biocides in the desired concentrations to the sterile mineral salts-glucose medium, the fungal spore suspension was added (0.1 ml of inoculum). The final spore concentration was approximately 10.sup.6 cfu/mL. The inoculated media was incubated at 28.degree. C. for 10-14 days.

B. Bacterial Evaluation

Nutrient broth growth medium was used. To prepare the medium, the following ingredients were added to one liter of deionized water: 8.0 g NaCl, 1.0 g glucose, and 1.0 g tryptone. The pH of the medium was adjusted to 6 with 1N HCL. This was distributed in 5 mL amounts in test tubes and autoclaved for 20 minutes at 121.degree. C. After addition of the biocides in the desired concentrations to the nutrient broth, 100 microliters of a suspension of Pseudomonas aeruginosa cells of approximately 10.sup.6 cfc/mL were added and incubated at 37.degree. C. for 5-7 days.

In the Examples, a synergistic effect was demonstrated in separate experiments by testing 3,5,-dimethyl-1-hydroxymethylpyrazole, designated as component A and the corresponding preservative or its salt as component B in a series of tests in varying ratios and a range of concentrations against the fungus, Aspergillus niger and also against the bacterium, Pseudomonas aeruginosa using the methods described above.

The lowest concentration of each mixture or compound which completely prevented growth of the fungus for about two weeks (10-14 days) and the bacterium for about one week (5-7 days) was taken as the end points for synergism calculations. End points for the various mixtures were then compared with the end points for the pure active ingredients alone in concomitantly prepared flasks or test tubes.

Synergism was demonstrated by the method described by Kull, E. C., Eisman, P. C., Sylwestrwicz, H. D., and Mayer, R. L. 1961. Applied Microbiology. 9:538-541 wherein

______________________________________QA/Qa + QB/Qb is less than 1______________________________________Qa (Busan .RTM. 1504) = Concentration of compound A in parts per million, acting alone, which produced an end point.Qb (preservative) = Concentration of compound B in parts per million, acting alone, which produced an end point.QA (Busan .RTM. 1504) = Concentration of compound A in parts per million, in the mixture, which produced an end point.QB (preservative) = Concentration of compound B in parts per million, in the mixture, which produced an end point.______________________________________

When the sum of QA/Qa and QB/Qb is greater than one, antagonism is indicated and when the sum is equal to one, addivity is indicated. When the sum of this value is less than one, synergism exists.

This procedure for demonstrating synergism of the compositions of this invention is a widely used and acceptable procedure. More detailed information is provided in the article by Kull et al. Further information concerning this procedure is contained in U.S. Pat. No. 3,231,509, which is incorporated herein by reference.

In general, however, an effective fungicidal and bactericidal response can be obtained when the synergistic combination is employed in concentrations ranging from about 0.1 to about 10,000 ppm of a 1-hydroxymethylpyrazole, preferably about 0.1 to about 5,000 ppm, and most preferably about 1 to about 2,000 ppm, and from about 0.1 to about 10,000 ppm of a preservative, preferably about 0.1 to about 5,000 ppm and most preferably about 1 to about 3,000 ppm.

TABLE 1______________________________________Summary of Synergism in Antibacterial Test (vs Ps. aeruginosa)Combination Synergism SI Range*______________________________________BS1504 + BS1210 yes 0.48-0.75BS1504 + Kathon CG yes 0.58-0.75BS1504 + Dowicil 200 yes 0.79-0.93BS1504 + Propyl Paraben yes 0.67-0.83BS1504 + Methyl Paraben yes 0.68-0.92BS1504 + BS1506 yes 0.70-0.97BS1504 + Sodium Omadine yes 0.68-0.98BS1504 + Germall II yes 0.73-0.98BS1504 + Germall 115 yes 0.77-0.96BS1504 + Bronopol yes 0.75-0.96BS1504 + Glydant yes 0.75-0.96______________________________________ TABLE 2______________________________________Summary of Synergism in Antifungal Test (vs A. niger)Combination Synergism SI Range______________________________________BS1504 + BS1210 yes 0.21-0.72BS1504 + Kathon CG yes 0.79-0.96BS1504 + Dowicil 200 yes 0.55-0.77BS1504 + Propyl Paraben no --BS1504 + Methyl Paraben no --BS1504 + BS1506 yes 0.67-0.93BS1504 + Sodium Omadine yes 0.34-0.77BS1504 + Germall II yes 0.73-0.97BS1504 + Germall 115 no --BS1504 + Bronopol yes 0.69-0.93BS1504 + Glydant yes 0.68-0.97______________________________________ *Sl = 1 (additive) or SI > 1 (antagonistic) were not included. TABLE 3______________________________________Combination of Busan 1504 with Busan 1210vs Ps. aeruginosaBS1504 BS1210 BS1504 BS1210(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________10 0 + 30 1 +20 0 + 30 2 +40 0 + 30 5 +80 0 + 30 10 +100 0 + 30 20 +120 0 - 40 1 +140 0 - 40 2 +160 0 - 40 5 +180 0 - 40 10 +200 0 - 40 20 - 0.58 0 10 + 50 1 + 0 20 + 50 2 + 0 30 + 50 5 - 0.48 0 40 + 50 10 - 0 50 + 50 20 - 0 60 + 60 1 + 0 80 - 60 2 + 0 100 - 60 5 - 0.5610 2 + 60 10 -10 5 + 60 20 -10 10 + 70 1 +10 20 + 70 2 +20 2 + 70 5 - 0.6520 5 + 70 10 -20 10 + 70 20 -20 20 +______________________________________ TABLE 4______________________________________Combination of Busan 1504 with Busan 1210vs A. nigerBS1504 BS1210 BS1504 BS1210(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________300 0 + 100 0.2 +400 0 + 100 0.4 +500 0 + 100 0.6 +600 0 - 100 0.8 - 0.27700 0 - 100 1.0 -800 0 - 100 1.5 -0 0.2 + 100 2.0 -0 0.5 + 200 0.1 +0 0.8 + 200 0.2 +0 1 + 200 0.4 - 0.380 2 + 200 0.6 -0 4 + 200 0.8 -0 8 - 200 1.0 -0 16 - 200 1.5 -0 0.2 + 300 0.1 - 0.5110 0.5 + 300 0.2 -10 0.8 + 300 0.4 -10 1.0 + 300 0.6 -10 1.5 + 300 0.8 -10 2.0 - 0.27 300 1.0 -10 4.0 - 300 1.5 -50 0.2 + 400 0.05 - 0.6750 0.5 + 400 0.1 -50 0.8 + 400 0.2 -50 1.0 - 0.21 400 0.4 -50 1.5 - 400 0.6 -50 2.0 - 400 0.8 -50 4.0 - 400 1.0 - 400 1.5 -______________________________________ TABLE 5______________________________________Combination of Busan 1504 with Kathon CGvs Ps. aeruginosaBS1504 KN CG BS1504 KN CG(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________10 0 + 30 1 +20 0 + 30 2 +40 0 + 30 5 +80 0 + 30 10 - 0.75100 0 + 30 20 -120 0 - 40 1 +140 0 - 40 2 +160 0 - 40 5 - 0.58180 0 - 40 10 -200 0 - 40 20 - 0 10 + 50 1 + 0 20 - 50 2 + 0 30 - 50 5 - 0.67 0 40 - 50 10 - 0 50 - 50 20 - 0 60 - 60 1 + 0 80 - 60 2 + 0 100 - 60 5 - 0.7510 2 + 60 10 -10 5 + 60 20 -10 10 + 70 1 +10 20 - 70 2 - 0.6820 2 + 70 5 -20 5 + 70 10 -20 10 - 0.67 70 20 -20 20 -______________________________________ TABLE 6______________________________________Combination of Busan 1504 with Kathon CGvs A. nigerBS1504 KN CG BS1504 KN CG(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________300 0 + 200 5 +400 0 + 200 10 +500 0 + 200 15 +600 0 + 200 20 +700 0 + 200 25 - 0.87800 0 - 200 30 - 0 10 + 200 35 - 0 20 + 300 5 + 0 30 + 300 10 + 0 40 - 300 15 + 0 50 - 300 20 - 0.87 0 60 - 300 25 - 0 70 - 300 30 - 0 80 - 300 35 - 10 10 + 400 1 + 10 20 + 400 2 + 10 30 + 400 4 + 10 40 + 400 8 + 50 10 + 400 16 - 0.90 50 20 + 400 20 - 50 30 + 400 25 - 50 40 - 400 30 -100 10 + 500 1 +100 20 + 500 2 +100 30 - 0.87 500 4 +100 40 - 500 8 + 500 16 - 500 20 - 500 25 -______________________________________ TABLE 7______________________________________Combination of Busan 1504 with Dowicil 200vs Ps. aeruginosaBS1504 DW200 BS1504 DW200(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________10 0 + 30 10 +20 0 + 30 20 +40 0 + 30 30 +80 0 - 30 40 +100 0 - 30 50 +120 0 - 30 60 +140 0 - 30 70 +160 0 - 40 10 +180 0 - 40 20 +200 0 - 40 20 + 0 100 + 40 40 + 0 110 + 40 50 + 0 120 + 40 60 + 0 130 + 50 10 + 0 140 + 50 20 + 0 150 + 50 30 - 0.79 0 160 + 50 40 + 0 180 - 50 50 - 0.9010 30 + 50 60 -10 40 + 60 10 +10 50 + 60 20 - 0.8610 60 + 60 30 -10 70 + 60 40 -20 20 + 60 50 -20 30 + 60 60 +20 40 + 70 10 - 0.9320 50 + 70 20 -20 60 + 70 30 -20 70 + 70 40 -______________________________________ TABLE 8______________________________________Combination of Busan 1504 with Dowicil 200vs A. nigerBS1504 DW200 BS1504 DW200(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________300 0 + 200 20 +400 0 + 200 50 +500 0 + 200 80 - 0.55600 0 + 200 100 -700 0 - 200 120 -800 0 - 200 150 - 0 100 + 200 200 - 0 200 + 200 250 - 0 300 - 200 300 - 0 400 - 300 10 + 0 500 - 300 20 + 0 600 - 300 40 + 10 50 + 300 60 - 0.63 10 100 + 300 80 - 10 200 - 0.68 300 100 - 10 250 - 300 120 - 10 300 - 300 150 - 50 50 + 300 200 - 50 100 + 400 10 + 50 150 + 400 20 + 50 200 - 0.74 400 40 + 50 250 - 400 60 - 0.77 50 300 - 400 80 -100 50 + 400 100 -100 100 + 400 120 -100 150 - 0.64 400 150 -100 200 - 400 200 -100 250 -100 300 -______________________________________ TABLE 9______________________________________Combination of Busan 1504 and Propyl Parabenvs Ps. aeruginosaBS1504 PPB BS1504 PPB(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________40 0 + 40 500 +60 0 + 40 600 +80 0 + 40 700 - 0.80100 0 + 40 800 -120 0 - 50 200 +140 0 - 50 300 +160 0 - 50 400 +180 0 - 50 500 - 0.75 0 200 + 50 600 - 0 400 + 50 700 - 0 600 + 60 100 + 0 800 + 60 200 + 0 900 + 60 300 + 0 1000 + 60 400 - 0.77 0 1200 + 60 500 - 0 1500 - 60 600 - 0 2000 - 70 50 +10 700 + 70 100 +10 800 + 70 200 - 0.7210 900 + 70 300 -20 600 + 70 400 -20 700 + 70 500 -20 800 + 80 10 - 0.6720 900 + 80 20 -30 500 + 80 40 -30 600 + 80 60 -30 700 - 0.72 80 80 -30 800 - 80 100 - 80 200 - 80 300 -______________________________________ TABLE 10______________________________________Combination of Busan 1504 with Propyl Parabenvs A. nigerBS1504 PPB BS1504 PPB(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________300 0 + 200 10 +400 0 + 200 20 +500 0 + 200 40 +600 0 + 200 60 +700 0 - 200 80 +800 0 - 200 100 + 0 50 + 200 150 + 0 100 + 300 10 + 0 150 + 300 20 + 0 200 - 300 40 + 0 250 - 300 60 + 0 300 - 300 80 + 0 400 - 300 100 + 0 500 - 400 10 + 10 50 + 400 20 + 10 80 + 400 40 + 10 100 + 400 60 + 10 150 + 400 80 + 50 50 + 400 100 + 50 80 + 50 100 + 50 150 +100 10 +100 20 +100 40 +100 60 +100 80 +100 100 +100 150 +______________________________________ TABLE 11______________________________________Combination of Busan 1504 and Methyl Parabenvs Ps. aeruginosaBS1504 MPB BS1504 MPB(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________40 0 + 30 400 +60 0 + 30 500 - 1.0780 0 + 30 600 -100 0 + 30 700 -120 0 - 30 800 -140 0 - 40 300 +160 0 - 40 400 +180 0 - 40 500 - 1.15 0 200 + 40 600 - 0 400 + 40 700 - 0 500 + 50 300 + 0 600 - 50 400 + 0 700 - 50 500 + 0 800 - 50 600 + 0 900 - 50 700 - 1.56 0 1000 - 60 200 + 0 1500 - 60 300 +10 400 + 60 400 +10 500 + 60 500 +10 600 - 1.08 60 600 - 1.5010 700 - 70 50 +10 800 - 70 100 +10 900 - 70 200 +20 400 + 70 300 +20 500 + 70 400 +20 600 - 1.16 70 500 - 1.4120 700 - 80 10 +20 800 - 80 20 + 80 50 + 80 100 + 80 200 + 80 300 +______________________________________ TABLE 12______________________________________Combination of Busan 1504 with Methyl Paraben vs A. nigerBS1504 MPB BS1504 MPB(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________300 0 + 200 100 +400 0 + 200 200 +500 0 + 200 400 +600 0 + 200 600 +700 0 + 200 800 - 1.25800 0 - 300 50 + 0 200 + 300 100 + 0 400 + 300 200 + 0 600 + 300 400 + 0 800 - 300 600 + 0 1000 - 300 800 - 1.37 0 2000 - 400 50 + 10 200 + 400 100 + 10 400 + 400 200 + 10 600 + 400 400 + 10 800 - 1.01 400 600 - 1.25 10 1000 - 400 800 + 50 200 + 500 10 + 50 400 + 500 50 + 50 600 + 500 100 + 50 800 - 1.06 500 200 + 50 1000 - 500 400 +100 100 + 500 600 - 1.37100 200 + 600 10 -100 400 + 600 50 +100 600 + 600 100 +100 800 - 1.13 600 200 + 600 400 + 600 600 - 1.50______________________________________ TABLE 13______________________________________Combination of Busan 1504 with Busan 1506 vs Ps. aeruginosaBS1504 BS1506 B51304 BS1506(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________40 0 + 20 10 +60 0 + 20 20 +80 0 + 20 30 +100 0 + 20 40 +120 0 + 20 50 +140 0 - 20 60 +160 0 - 20 70 - 0.73180 0 - 30 5 + 0 20 + 30 10 + 0 40 + 30 20 + 0 60 + 30 30 + 0 80 + 30 40 + 0 100 + 30 50 + 0 120 - 30 60 + 0 140 - 40 1 + 0 160 - 40 5 + 0 180 - 40 10 + 0 200 - 40 20 + 5 10 + 40 30 + 5 20 + 40 40 + 5 30 + 40 50 + 5 40 + 50 1 + 5 50 + 50 5 + 5 60 + 50 10 + 5 70 + 50 20 + 5 80 - 0.70 50 30 +10 20 + 50 40 +10 30 +10 40 +10 50 +10 60 +10 70 +10 80 +______________________________________ TABLE 14______________________________________Combination of Busan 1504 with Busan 1506 vs A. nigerBS1504 BS1506 BS1504 BS1506(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________300 0 + 200 50 +400 0 + 200 100 +500 0 + 200 150 +600 0 + 200 200 +700 0 - 200 250 - 0.91800 0 - 200 300 - 0 100 + 300 20 + 0 200 + 300 40 + 0 300 + 300 60 + 0 400 - 300 80 + 0 500 - 300 100 + 0 600 - 300 150 + 10 100 + 300 200 - 0.93 10 200 + 300 250 - 10 300 - 0.76 300 300 - 10 400 - 400 20 + 50 100 + 400 40 + 50 200 + 400 60 + 50 300 - 0.82 400 80 + 50 400 - 400 100 +100 50 + 400 150 +100 100 + 400 200 -100 150 + 400 250 -100 200 + 400 300 -100 250 - 0.77100 300 -100 400 -______________________________________ TABLE 15______________________________________Combination of Busan 1504 with Sodium Omadinevs P. aeruginosaBS1504 SOMD BS1504 SOMD(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________40 0 + 40 10 +60 0 + 40 20 +80 0 + 40 40 +100 0 + 40 60 - 0.93120 0 - 40 80 -140 0 - 40 100 -160 0 - 50 10 +180 0 - 50 20 + 0 20 + 50 40 - 0.82 0 40 + 50 60 - 0 60 + 50 80 - 0 80 + 50 100 - 0 100 - 60 10 + 0 120 - 60 20 + 0 150 - 60 40 - 0.90 0 200 - 60 60 - 0 300 - 60 80 -10 40 + 60 100 -10 60 + 70 10 - 0.6810 80 + 70 20 -10 100 - 70 30 -20 20 + 70 40 -20 40 + 70 50 -20 60 - 0.76 70 60 -20 80 - 80 10 - 0.7720 100 - 80 20 -30 20 + 80 30 -30 40 + 80 40 -30 60 - 0.85 80 50 -30 80 - 80 60 -30 100 -______________________________________ TABLE 16______________________________________Combination of Busan 1504 and Sodium Omadinevs A. nigerBS1504 SOMD BS1504 SOMD(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________300 0 + 200 0.2 +400 0 + 200 0.5 +500 0 + 200 1 +600 0 + 200 2 +700 0 + 200 4 - 0.65800 0 - 200 8 - 0 1 + 200 16 - 0 5 + 300 0.2 + 0 10 - 300 0.5 + 0 20 - 300 1 + 0 30 - 300 2 + 0 50 - 300 4 - 0.77 0 80 - 300 8 - 0 160 - 300 16 -10 1 + 400 0.1 +10 5 + 400 0.2 +10 10 - 400 0.4 +10 20 - 400 0.8 +50 1 + 400 1 +50 5 + 400 2 - 0.7050 10 - 400 4 -50 20 - 400 8 -100 0.5 + 400 16 -100 1 + 500 0.1 +100 2 + 500 0.2 +100 4 - 0.53 500 0.4 +100 8 - 500 0.8 +100 16 - 500 1 - 0.72 500 2 - 500 4 - 500 8 - 500 16 -______________________________________ TABLE 17______________________________________Combinaition of Busan 1504 and Germall IIvs P. aeruginosaBS1504 GII BS1504 GII(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________40 0 + 40 50 +60 0 + 40 80 +80 0 + 40 100 -100 0 + 40 120 -120 0 - 40 140 -140 0 - 40 160 -160 0 - 50 20 +180 0 - 50 40 + 0 50 + 50 80 - 0.98 0 80 + 50 100 - 0 100 + 50 120 - 0 120 + 50 140 - 0 140 - 60 20 + 0 160 - 60 40 + 0 180 - 60 80 - 0 200 - 60 100 -10 100 + 60 120 -10 120 - 0.94 60 140 -10 140 - 70 20 +10 160 - 70 40 - 0.8710 180 - 70 60 -20 100 + 70 80 -20 120 - 70 100 -20 140 - 70 120 -20 160 - 70 140 -20 180 - 80 10 +30 50 + 80 20 - 0.8130 80 + 80 40 -30 100 - 0.96 80 50 -30 120 - 80 60 -30 140 - 80 80 -30 160 - 80 100 -______________________________________ TABLE 18______________________________________Combination of Busan 1504 with Germall IIvs . A. nigerBS1504 Germl II BS1504 Germl II(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________300 0 + 200 250 +400 0 + 200 500 +500 0 + 200 1000 +600 0 + 200 1500 - 0.85700 0 + 200 2000 -800 0 - 300 250 + 0 500 + 300 500 + 0 1000 + 300 1000 - 0.77 0 1500 + 300 1500 - 0 2000 + 300 2000 - 0 2500 - 400 100 + 0 3000 - 400 250 + 10 500 + 400 500 + 10 1000 + 400 1000 - 0.90 10 1500 + 400 1500 - 10 2000 + 400 2000 - 10 2500 - 500 100 + 50 500 + 500 250 + 50 1000 + 500 500 + 50 1500 + 500 1000 - 50 2000 - 0.86 500 1500 - 50 2500 - 600 100 +100 500 + 600 250 +100 1000 + 600 500 - 0.95100 1500 - 0.73 600 1000 -100 2000 - 600 1500 - 700 100 + 700 250 + 700 500 - 700 1000 - 700 1500 -______________________________________ TABLE 19______________________________________Combination of Busan 1504 and Germall 115vs P. aeruginosaBS1504 G115 BS1504 G115(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________40 0 + 30 100 +60 0 + 30 200 +80 0 + 30 300 +100 0 + 30 400 -120 0 + 30 500 -140 0 - 30 600 -160 0 - 40 50 +180 0 - 40 100 + 0 100 + 40 200 + 0 200 + 40 300 - 0.88 0 300 + 40 400 - 0 400 + 40 500 - 0 500 - 50 50 + 0 600 - 50 100 +10 100 + 50 200 +10 200 + 50 300 - 0.9610 300 + 50 400 -10 400 + 50 500 -10 500 - 60 50 +10 600 - 60 100 +20 100 + 60 200 - 0.8320 200 + 60 300 -20 300 + 60 400 -20 400 - 0.94 60 500 -20 500 - 70 20 +20 600 - 70 50 + 70 100 + 70 200 - 0.90 70 300 - 80 20 + 80 50 + 80 100 - 0.77 80 200 - 80 300 -______________________________________ TABLE 20______________________________________Combination of Busan 1504 with Germall 115vs A. nigerBS1504 Germ 115 BS1504 Germ 115(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________300 0 + 200 1000 +400 0 + 200 2000 +500 0 + 200 3000 +600 0 + 200 4000 +700 0 + 200 5000 +800 0 - 200 6000 + 0 2000 + 200 7000 - 1.05 0 4000 + 300 1000 + 0 5000 + 300 2000 + 0 6000 + 300 3000 + 0 8000 - 300 4000 + 0 10000 - 300 5000 - 1.00 10 2000 + 300 6000 - 10 4000 + 400 1000 + 10 5000 + 400 2000 + 10 6000 + 400 3000 + 10 7000 + 400 4000 - 1.00 10 8000 + 400 5000 - 50 2000 + 400 6000 - 50 4000 + 500 1000 + 1.03 50 5000 + 500 2000 + 50 6000 + 500 3000 - 1.00 50 7000 + 500 4000 - 50 8000 - 1.06 500 5000 -100 2000 + 500 6000 -100 4000 +100 5000 +100 6000 +100 7000 +100 8000 - 1.12______________________________________ TABLE 21______________________________________Combination of Busan 1504 and Bronopolvs . P. aeruginosaBS1504 Bronopol BS1504 Bronopol(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________40 0 + 30 0.5 +60 0 + 30 1 +80 0 + 30 2 - 0.75100 0 - 30 3 -120 0 - 30 4 -140 0 - 30 5 -160 0 + 40 0.5 +180 0 + 40 1 + 0 0.5 + 40 2 - 0.83 0 1 + 40 3 - 0 2 + 40 4 - 0 3 + 40 5 - 0 4 - 50 0.2 + 0 5 - 50 0.5 + 0 8 - 50 1 + 0 10 - 50 2 - 0.9210 0.5 + 50 3 -10 1 + 50 4 -10 2 + 60 0.2 +10 3 - 0.83 60 0.5 +10 4 - 60 1 +10 5 - 60 2 -20 0.5 + 60 3 -20 1 + 60 4 -20 2 + 70 0.2 +20 3 - 0.92 70 0.5 +20 4 - 70 1 - 0.8320 5 - 70 2 - 70 3 - 80 0.2 + 80 0.5 + 80 1 - 0.92 80 2 - 80 3 -______________________________________ TABLE 22______________________________________Combination of Busan 1504 with Bronopolvs . A. nigerBS1504 Bronopol BS1504 Bronopol(ppm) (ppm) (ppm) (ppm) Growth SI______________________________________300 0 + 200 100 +400 0 + 200 200 +500 0 + 200 300 +600 0 + 200 400 - 0.86700 0 - 200 500 -800 0 - 200 600 - 0 300 + 200 700 - 0 400 + 300 50 + 0 500 + 300 100 + 0 600 + 300 200 + 0 700 - 300 300 - 0.86 0 800 - 300 400 - 0 1000 - 300 500 -10 300 + 300 600 -10 400 + 300 700 -10 500 + 400 50 +10 600 - 0.87 400 100 +10 700 - 400 200 - 0.8610 800 - 400 300 -50 300 + 400 400 -50 400 + 400 500 -50 500 + 400 600 -50 600 - 0.93 500 20 +50 700 - 500 50 +50 800 - 500 100 - 0.86100 100 + 500 200 -100 200 + 500 300 -100 300 + 500 400 -100 400 + 500 500 -100 500 - 0.86 600 20 +100 600 - 600 50 +100 700 - 600 100 - 600 200 - 600 300 - 600 400 -______________________________________ TABLE 23______________________________________Combination of Busan 1504 with Glydantvs P. aeruginosaBS1504 Glydant BS1504 Glydant(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________40 0 + 30 100 +60 0 + 30 200 +80 0 + 30 300 - 0.96100 0 + 30 400 -120 0 + 30 500 -140 0 - 30 600 -160 0 - 40 50 +180 0 - 40 100 + 0 100 + 40 200 + 0 200 + 40 300 - 0 300 + 40 400 - 0 400 - 40 500 - 0 500 - 50 25 + 0 600 - 50 50 +10 100 + 50 100 +10 200 + 50 200 - 0.8610 300 - 0.82 50 300 -10 400 - 50 400 -10 500 - 60 25 +10 600 - 60 50 +20 100 + 60 100 +20 200 + 60 200 - 0.9320 300 - 0.89 60 300 -20 400 - 60 400 -20 500 - 70 10 +20 600 - 70 25 + 70 50 + 70 100 - 0.75 70 200 - 70 300 - 80 10 + 80 25 + 80 50 + 80 100 + 80 200 + 80 300 -______________________________________ TABLE 24______________________________________Combination of Busan 1504 with Glydantvs A. nigerBS1504 Glydant BS1504 Glydant(ppm) (ppm) Growth SI (ppm) (ppm) Growth SI______________________________________300 0 + 200 250 +400 0 + 200 500 +500 0 + 200 1000 - 0.68600 0 + 200 1500 -700 0 - 200 2000 -800 0 - 300 250 + 0 500 + 300 500 + 0 1000 + 300 1000 - 0.83 0 1500 + 300 1500 - 0 2000 + 300 2000 - 0 2500 - 400 100 + 0 3000 - 400 250 + 10 500 + 400 500 + 10 1000 + 400 1000 - 0.97 10 1500 + 400 1500 - 10 2000 + 400 2000 - 10 2500 - 500 100 + 10 3000 - 500 250 + 50 500 + 500 500 - 0.91 50 1000 + 500 1000 - 50 1500 + 500 1500 - 50 2000 - 0.87 600 50 + 50 2500 - 600 100 + 50 3000 - 600 250 +100 500 + 600 500 -100 1000 + 600 1000 -100 1500 + 700 50 +100 2000 - 0.94 700 100 +100 2500 - 700 250 -100 3000 - 700 50 - 700 1000 -______________________________________

Claims

1. A composition comprising

(a) a compound having the formula (I) ##STR3## where R and R' are independently selected from hydrogen and alkyl of 1 to 4 carbon atoms, R" is a moiety selected from the group consisting of (a) hydrogen, (b) a halogen atom selected from the group consisting of F, Cl, Br, and I, and (c) a nitro group, or the hydrochloride salt of said compound; and

(b) at least one preservative selected from hexahydro-1,3,5-tris(2-hydroxyethyl)-s-triazine; bis 1,4-(bromoacetoxy)-2-butene; mixture of 5-chloro-2-methyl-4-iso-thiazoline-3-one and 2-methyl-4-iso-thiazoline-3-one; or C.sub.1 -C.sub.6 alkyl hydroxybenzoate, wherein said components (a) and (b) are present in a combined amount synergistically effective to control the growth of a bacterium, fungus, or a mixture thereof.

2. The composition of claim 1, wherein said compound of formula I is 3,5-dimethyl-1-hydroxymethylpyrazole.

3. The composition of claim 1, wherein said bacterium or fungus is Aspergillus niger or Pseudomonas aeruginosa.

4. The composition of claim 1, wherein the weight ratio of (a) to (b) is from about 0.1:99 to about 99:0.1.

5. The composition of claim 4, wherein the weight ratio of (a) to (b) is from about 1:30 to about 30:1.

6. The composition of claim 5, where the weight ratio of (a) to (b) is from about 1:5 to about 5:1.

7. The composition of claim 1, wherein said preservative is hexahydro-1,3,5-tris(2-hydroxyethyl)-s-triazine.

8. The composition of claim 1, wherein said preservative is bis-1,4-(bromoacetoxy)-2-butene.

9. The composition of claim 1, wherein said preservative is a mixture of 5-chloro-2-methyl-4-iso-thiazoline-3-one and 2-methyl-4-iso-thiazoline-3-one.

10. The composition of claim 1, wherein said preservative is C.sub.1 -C.sub.6 alkyl hydroxybenzoate.

11. A method of controlling the growth of at least one microorganism selected from a bacterium, fungus, or a mixture thereof, in or on a material or medium susceptible to attack by the microorganism comprising the step of adding to the material or medium a composition comprising

(a) a compound having the formula (I) ##STR4## where R and R' are independently selected from hydrogen and alkyl of 1 to 4 carbon atoms, R" is a moiety selected from the group consisting of (a) hydrogen, (b) a halogen atom selected from the group consisting of F, Cl, Br, and I, and (c) a nitro group, or the hydrochloride salt of said compound; and

(b) at least one preservative selected from hexahydro-1,3,5-tris(2-hydroxyethyl)-s-triazine; bis 1,4-(bromoacetoxy)-2-butene; mixture of 5-chloro-2-methyl-4-iso-thiazoline-3-one and 2-methyl-4-iso-thiazoline-3-one; or C.sub.1 -C.sub.6 alkyl hydrobenzoate, wherein said components (a) and (b) are present in a combined synergistically effective amount to control said growth.

12. The method of claim 11, wherein said compound of formula I is 3,5,-dimethyl-1-hydroxymethyl pyrazole.

13. The composition of claim 11, wherein said bacterium or fungus is Aspergillus niger or Pseudomonas aeruginosa.

14. The method of claim 11, wherein said material or medium is wood pulp, wood chips, wood, lumber, paints, dyes, leathers, adhesives, coatings, animal hides, tanning liquors, paper mill liquors, metalworking fluids, starch, petrochemicals, acrylic latex paint emulsions, pharmaceutical formulations, cooling tower water, cosmetics and toiletry formulations, textiles, influent and effluent plant waters, waste water, geological drilling lubricants or agrochemical compositions.

15. The method of claim 12, wherein the material or medium is in the form of a solid, dispersion, emulsion or solution.

16. The method of claim 11, wherein said components (a) and (b) can be added separately to the material or medium.

17. The method of claim 11, wherein said preservative is hexahydro-1,3,5-tris(2-hydroxyethyl)-s-triazine.

18. The method of claim 11, wherein said preservative is bis-1,4-(bromoacetoxy)-2-butene.

19. The method of claim 11, wherein said preservative is a mixture of 5-chloro-2-methyl-4-iso-thiazoline-3-one and 2-methyl-4-iso-thiazoline-3-one.

20. The method of claim 11, wherein said preservative is C.sub.1 -C.sub.6 alkyl hydroxybenzoate.

21. A method for preventing spoilage of a material or medium caused by bacteria or fungi comprising the step of adding to a material or medium a composition comprising

(a) a compound having the formula (I) ##STR5## where R and R' are independently selected from hydrogen and alkyl of 1 to 4 carbon atoms, R" is a moiety selected from the group consisting of (a) hydrogen, (b) a halogen atom selected from the group consisting of F, Cl, Br, and I, and (c) a nitro group, or the hydrochloride salt of said compound; and

(b) at least one preservative selected from hexahydro-1,3,5-tris(2-hydroxyethyl)-s-triazine; bis 1,4-(bromoacetoxy)-2-butene; mixture of 5-chloro-2-methyl-4-iso-thiazoline-3-one and 2-methyl-4-iso-thiazoline-3-one; or C.sub.1 -C.sub.6 alkyl hydroxybenzoate, wherein said components (a) and (b) are present in a combined synergistically effective amount to prevent or mitigate said spoilage.

22. The method of claim 21, wherein said compound of formula I is 3,5,-dimethyl-1-hydroxymethyl pyrazole.