AU1836099A

AU1836099A - Synergistic antimicrobial compositions containing methylene-bis(thiocyanate) and an organic acid

Info

Publication number: AU1836099A
Application number: AU18360/99A
Authority: AU
Inventors: Vanja M King; David Oppong
Original assignee: Buckman Laboratories International Inc
Current assignee: Buckman Laboratories International Inc
Priority date: 1994-01-27
Filing date: 1999-02-22
Publication date: 1999-04-29

Description

Regulation 3.2(2)

AUSTRALIA

Patents Act 1990

ORIGINAL

COMPLETE SPECIFICATION STANDARD PATENT Application Number: Lodged: Invention Title: SYNERGISTIC ANTIMICROBIAL COMPOSITIONS CONTAINING METH4YLENE-BIS(THIOCYANATE) AND AN ORGANIC ACID The following statement is a full description of this invention, including the best method of performing it known to us SYNERGISTIC ANTIMICROBIAL COMPOSITIONS CONTAINING METYLENE-BIS(THIOCYANATE) AND AN ORGANIC ACID SThe present invention relates to certain compositions and processes useful for controlling the growth of one or more microorganisms and for preventing spoilage caused by Sbacteria and fungi in various products, materials or media, particularly industrial products, materials or media. These products, materials or media include paints, leather, wood, wood pulp, wood chips, starch, proteinacecus materials, lumber, animal hides, vegetable tanning liquors, cosmetics, toiletry formulations, acrylic latex paint emulsions, adhesives, coatings, metalworking fluids, cooling tower water, textiles, petrochemicals, pharmaceutical formulations, geological drilling lubricants, paper mill liquors, and agrochemical compositions.

*'The novel compositions and processes incorporating the composition of the present invention show unexpected, synergistic activity against microorganisms, including S bacteria and fungi. Specifically, the invention is directed to the use of compositions and processes containing methylene-bis(thiocyanate) and at least one organic acid, its Ssalt or ester.

Many of the products, materials or media referred to above, when wet or subjected to treatment in water, are susceptible to bacterial and/or fungal deterioration or degradation unless steps are taken to inhibit such degradation or deterioration. To control deterioration or degradation caused by microorganisms, various industrial microbicides are used, but some of these biocides are of questionable utility because they have undesirable odors, are high in cost, show a low degree of effectiveness and/or create hazards with respect to storage, use and/or handling.

For instance, the use of such popular industrial microbicides as organomercury compounds, organotin compounds and chlorinated phenols has come under increasing regulatory i\; -2 pressure in recent times because of their high toxicity and concern about their adverse effects on the environment.

Consequently, the industry has continued to seek improved biocides that have low toxicity and are capable of exhibiting prolonged biocidal effect at normal use levels.

Organic acids can be used alone to control microorganisms, and while some of these compounds are generally regarded as safe, many of them have low efficacy against bacteria and fungi unless extremely high concentrations are used. In excessive concentrations, these organic acids can be expensive and could even be corrosive to certain industrial materials. A method that can prevent excessive use of organic acids and hence decrease cost is therefore desirable.

Alternatively, methylene-bis(thiocyanate) can be used alone in low concentrations as low toxicity biocides.

However, at low concentrations, methylene-bis(thiocyanate) tends to have a narrow antimicrobial spectrum and fails to completely prevent the growth of microorganisms.

Accordingly, the present invention is directed to microbicidal compositions and processes incorporating these compositions that substantially obviate one or more of the problems, limitations, and disadvantages of the prior art.

In particular, the compositions of the present invention are capable of controlling the growth of at least one microorganism, especially fungi or bacteria, over prolonged -periods of time, and are safe and economical to use. The present invention also is directed to methods or processes of controlling the growth of at least one microorganism.

The present invention provides a composition to control the growth of at least one microorganism comprising synergistically effective amounts of methylene-bis(thiocyanate) and at least one organic acid, its salt or ester. The composition provides superior a c i -3microbicidal activity at low concentrations against a wide range of microorganisms.

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 microqrganism which comprises the step of adding to the material or medium a composition of the present invention, where the components of the composition are present in synergistically effective amounts.

Furthermore, the present invention provides a method of preventing spoilage of a product, material or medium caused by bacteria and/or fungi comprising the step of adding to the product, material or medium, a composition of the present invention where the components of the composition are present n synergistically effective amounts.

The synergistically effective amounts vary in accordance with the material or medium to be treated and can, for a particular application, be routinely determined without undue experimentation by one skilled in the art.

The present invention also embodies the separate Saddition of methylene-bis(thiocyanate) and at least one organic acid, its salt or ester 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 methylene-bis(thiocyanate) and at least one o'-ganic acid, its salt or ester present in the system at the time of use is that synergistically effective amount required to control the growth of at least one microorganism.

The compositions of the present invention are also I useful in preserving various types of industrial products, media or materials that are susceptible to attack by microorganisms. Such products, media or materials include but are not limited to dyes, pastes, lumber, leather, textiles, pulp, wood chips, tanning liquors, paper mill liquors, polymer emulsions, paints, paper and other coatinu i' and sizing agents, metalworking fluids, geological drilling lubricants, petrochemicals, cooling water systems, pharmaceutical formulations, cosmetics and toiletry formulations.

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

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

i. t is to be understood that both the foregoing general descriDtion and the following detailed 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 chemicals in the product would produce an additive (neutral) effect.

2) The chemicals in the product would produce an antagonistic (negative) effect, or 3) The chemicals in the product would produce a synergistic (positive) 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 advantage.

It is well-known in the microbicidal literature that there is no theoretical method to provide the likelihood of knowing, before actually testing, whether additive, E 5 antagonistic or synergistic effects: will be obtained when two biocides are mixed to yield a formulation.

IThe microbicidal compositions combining methylenebis(thiocyanate) and at least one organic acid, its salt or ester demonstrate an unexpected, synergistic effect compared to the respective components alone. Thus these comoositions achieve superior, i.e. greater than additive microbicidal activity at low concentrations against a wide variety of microorganisms. Examples of microorganisms include fungi and bacteria such as, but not limited to, Trichoderma hazianum and Pseudomonas aeruginosa. These two organisms are some of the most common organisms associated with spoilage of S products, materials or media. Since these two are also some of the toughest organisms to control, the comoosition of the present invention is believed to be effective against most bacteria and fungi. Preferably,-the compositions of the present invention have a low toxicity The preparation of methylene-bis(thiocvanate) (MTC) is described in U.S. Patent No. 3,524,871, which disclosure is fully incorporated by reference. MTC is commercially available and also easily synthesized from commercially available raw materials. A preferred MTC according to the present invention is 2-methylene-bis(thiocyanate).

The organic acids of the present invention are any organic acid that produce a synergistic effect when combined .with the MTC. Examples of organic acids include aromatic Sorganic acids, cyclic-organic acids, aliphatic organic acids, their salts or esters. Specific examples of effective organic acids according to the invention include dehydroacetic acid, octanoic acid, nonanoic acid, formic acid, sorbic acid, acetic acid, oxalic acid, glycolic acid, citric acid, gluconic acid, malic acid, propionic acid, lauric acid, undecylenic acid, benzoic acid or derivatives of benzoic acid such as 2 -hydroxybenzoic acid, 3 -hydroxybenzoic 6-t acid or 4-hydroxybenzoic acid, methyl paraben or propyl Daraben- The salts of the organic acids, preferably those containing calcium, zinc, potassium, or sodium, may be used, such as sodium benzoate and potassium sorbate. Preferred esters are parabens such as methyl paraben and propyl oaraben.

In accordance with the present invention, mixtures of these organic acids, salts or esters can also be used. When such mixtures are used in combination with MTC, at least one of the organic acids in the mixtures has a synergistic relationship with MTC. Organic acids, salts and esters useful in the invention are commercially available or may be synthesized from commercially available raw materials.

The organic acid may be chosen, for example, based on the comoatibility of the acid with the products, materials or media. The compatibility is readily determined by adding the organic acid to the products, materials or media to be used.

Compatibility may be determined by criteria such as S solubility in a fluid system and/or lack of reactivity with the fluid in question. When used in a fluid system, for examole, it is preferable that the organic acid be freely i'soluble or dispersible in the particular fluid system, resulting in a uniform solution or dispersion. Examples of fluid systems are tanning liquor, paper mill liquor, cooling tower water, and paints.

In accordance with the present invention, the composition may be in the form of a solid, dispersion, emulsion or solution, depending on the particular application- Further, the components of the composition may be applied separately or may be combined first and then applied to the product, material or medium.

?i The com~nosition of the Dresent- invention may be ureDared an jiou id orm by dissolvi nc comoonents and in an organic solvent.

in the following discussion of preferlred ermodiments, component is 2-methylen e-bis(thiocyanate) and comp~onent is at least one organic acidQ, its salt or ester.

As described above, comoonents and of the composition are used in syneroisticallv effective amounts.

The weight ratios of to vary depending on the type of-" microorganisms, as well as the products, materials or media to which the composition is applied. one skilled in the art can readily determine the appropriate weight ratiJos for a soecific application.

*in the present invention, the weight ratio of component to comoonent preferably ranges from about 0.01:99 to about 99:0.01, imore preferablyv from about 1:30 to about 30:1, *and most Dreferablv about 1:5 to about 5:1.

The following aooroximarte range of weight ratios of MTC to the fL'ollowing organic acids are also preferred according to the present invention: MTC: benzoic acid 1:5 to 0.1:100 -*MTC: sodium benzoate 1 *100 to 0.1:400 MTC: methyl paraben 1:5 to 0.01:100 MTC: Droovl1 oaraben 1:5 to 0.01:100 MTC: p-hydroxybenzoic acid 1:5 to 0.1:250 MTC: dehydroacetic acid 1:5 to 1:400 MTC: octanoic acid 1:5 to 1:400 MTC: rionaiioic acid 1:10 to 1:400 MTC: formic acid 1:50 to 1:800 MTC: sorbic acid 1:5 to 0.1:100 MTC: Potassium sorbate 1:5 to 1:800 MTC: acetic acid 1:100 to 1:200 MTC: oxalic acid 1:50 to 1:200 MTC: glycolic acid 1:100 to 1:400 MIT C: citric acid 1:100 to 1:-;00 MTC Malic acid 1:100 to 1:400 MTC: oronionic acid 1:10 to 1:400 MTC: lauric acid 1:5 to 0.1:400 MTC: undecylenic acid 1:5 to 0.'1:500 ±In general, however, an effective funqicidal and bactericidal response can be obtained when the synergistic combination is employed in concentrations ranging from about 0.01 to about 3000 ppm ofL rethylene-bis(thiocyanate), preferably from about 0.1. to about 1000 Dom, and most P referably from about 0.1 to about 500 Prm, anda from about.

0.1 PPM to about V~ ywigto h oraanic acid, D~referablv from about 0.1 to about 5000 ppm, and most preferably from about 0.1 to about 2000 Dn- The present invention also provides a method for controlling the growth ofZ at least one microorganism in or on a material or medium susceptible to attack by the mlicrooraanism which comprises the st~ep of aool ving to the material or medium a composition ofz the present -inverntion, where the components of the composition are present in Syner isticallNr efrective amounts.

Furthermore, the present invention provides a method of Preventing spoilage of a product, material or medium caused by bacteria and/or fungi comorisn the are ing to said Droduct, material or medium, a composition of the p~resent invention where the components of7 the composition are present in synergistically effective amounts. For example, the composition may be used to nrevent the spoilage of:! seeds or crops, e-Cr. cotton, barley, rice, maize, tobacco, etc.

The mode and rate of application of the comoosition varies depending upon thie intended use of the compnosition.

For instance, the composition may be applied by snrav~no or brushing onto a material or product. The material or podc could also be treated by dipaping in a suitable formulation orthe compoosition. For a licquid or liqu~.d-like medium, the commostiof could be added into the medium oy pourxina, Or bNy meteriflg with a suitable device, so tnat a solution or a d~soersion containing the coi-zpos2.tion is oroduce~o. Ift us ec.

as a l1O'Jid preservati.ve, -;or example, the composition maY be nrepared as an aarueous emulsion. Tznecessary, a surfactant may be added to the composition- In accordance with the invenzion, additional comoponents such as insecticides and the like -may be added to the foregoing preparations without atteccingl the synergistic effects of the composition. insecti~cides that may be used include but are not limited to pyrethrns, nicotine, chlordane, parathions, and methoxychlor.

The synergistic activity of the combinations described above has been confirmed using standard laboratory techniques, as illustrated below- The t-o1Thwiflo exampules are intended to illustrate, not limit, the scone of' toe present invention.

Microbioloaical evaluation A. Fungal evaluation A mineral salts-glucose medium was first prenared by adding to liter of deionized water: 0.7 a ofl KM.)PO, 0.7 c of mgSO 01 l.Og of NH NO.. 0.005 g NaCL, 0-002 9 44 eO71H 0, 0.002 a ZnSO 7H 0, 0-001 g MnSO 71: 0, and 10 c; FeO*24' 4* 42' of Glucose. The DI{ of the medium was adjusted to 6 with iN NaOH. The medium was then dispensed in 5 a amounts into test tubes and autoclaved at 121 0 C for 20 minutes.

The fungus, 7richodermuE harZianun. was grown on a potato dextrose agar slant for 7 to 10 days and a spore suspension prepared by washing dow-n the snores -t;ram the slant into a sterile saline solution. After the addition of biocides -4n the desired concentrations to toe sterile mi-neral saltsglucose medium, the fungal spore suspension was added. The 10 final snore concentration was approximately 10cfu/mL- The inoculated media was incubated at 28 0 C for 14 days.

B. Bacterial evaluation Nutrieit broth (2.5 g/liter of deionized water) was prepared and the pH adjusted to 6 with IN HCL. This was dispensed in 5 mL amounts into test tubes and autoclaved for minutes at 121 0 C. After the addition of biocides in the desired concentrations to the nutrient broth, 100 microliters or a suspension of Pseudomonas aeruginosa cells of approximately 9.3 x 10 8 cfc/mL were added and incubated at 37 0 C for 48 hours.

In Examples 1 through 19, a synergistic effect was demonstrated by testing the combination of methylenebis(thiocyanate), designated as component A, and the corresponding organic acid, its salt or ester, designated as component B, in a series of tests in varying ratios and a range of concentrations against the fungus Trichoderma harzianum and also against the bacterium Pseudomonas aeruganosa, using the methods described above.

For each component A and B in a mixture containing A and B and for each component A and B acting alone, the lowest concentration which completely prevented growth of the fungi for two weeks and the bacteria for 48 hours was determined.

These concentrations were used as end points for synergism calculations. End points for the components alone or in mixtures described above were then compared with the end points for the pure active ingredients alone in similarly prepared flasks or test tubes.

Synergism was demonstrated by the method described by Kull, Eisman, Sylwestrwicz, and Mayer, R.L.

1961. Applied Microbiology. 9: 538-541 wherein: 11 QA/Qa QB/Qb is less than 1 Qa Concentration of compound A in parts per million, acting alone, which produced an end point.

Qb Concentration of compound B in parts per million, acting alone, which produced ah end point.

QA Concentration of compound A in parts per million, in the mixture, which produced an end point.

QB Concentration of compound B in parts per million, in the mixture, which produced an end point.

When the sum bfe A/Qo.and -/Qb is greater than one, antagonism is indicated hen the sum is ecual to one, additivity is indicate thC sum of this value is less than one, synergism exir'tr.. This procedure for-de n trating.synergism of the compositions of this inveti n a widely used and accepted.

More detailed information is. provided in the article by Kull et. al. Further information concerning this procedure is contained in U.S. Patent No. 3,231,509, which disclosure is fully incorporated by reference.

The Examples demonstrate that in almost all of the samples, the combination of MTC with a corresponding organic acid produced a synergistic result (indicated by a ratio value of less than one.) There were a few samples, such as Table 2, <1.25 or where synergistic results were inconclusive because endpoints for the acids used alone were not determined.

It will be apparent to those skilled in the art that various modifications and variations can be made in the compositions and methods of the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention cover the modifications and variations of this invention provided they come within the scope of the appended claims and their equivalents.

'~EIWA* aaa~H-p--rr~aeraaas~5-~ap~p~~ ~blre

'K

12 Example I Compooent A MTC COMP~aa B B.-rzoic acid es roducing end poinr~s (prrn)

IQS/Q

Tixl oderna io0 I 2.5 250 0.25 0.5 0.75 5.0 25 0.5 .0.05 0.55 15.0 50 0-5 0.1 0.6 5. 100 0.5 0.2 0.7 50250 <0.5 <0.5 <1I 500 P-e1 1 n? I 0.1 25 0.1 0.1 0.2 0.1 50 J0.1 0.2 0.3 0.1 to10 0.1 0.4 0.25 25 0.25 0.1 0.35 0.25 s0 0.25 0.2 0.45 0.25 100 10.25 0.4 0.65 10 0 I:S 0.04 0.54 50 0.5 0.2 0.7 250 -13 Example 2 Component A MTC Component B Sodium benazoate- Test organism Quantities producind end points (ppm) [Trchoderma 10_ JQ J~ ,I 11 {0.5 05 2 50 10.5 F:2 5 0-75 0.5_ 0.5 1_ 0.5 _1 a

A

A4- -1

U

t .4 Example 3 Component A MTC Component B Sorbic acid Test organism Quantifie producing end points (ppm) TrcomL a o 10 Q!.

0.5 So150 fo.o 0.5 0.55 01 0.5 0.6 25250 0.25 0.25 500 10.25 10.5 0.75 125 1 0.5 0.03 0.53 50 0.5 0.05 0.55 5 100 0.5 0.1 0.6 5 250 0.5 0.25 0.75 5 1500 0.5 0.5 1 -1000 -J F PsudornonZ 1- OA 100 0.1 11 .4 0.25 100 0.25 0.4 0.65 0.5 10 0.5 0.04 0.54 0.5 25 0.5 0.1 0.6 0.5 so 5 0.5 0.2 0.7 0.5 100 0.5 j10.4 0.9 250 I

I

-4 Faample 4 Component A MTC Component B =potassium sorbate Test organism Quantities producing end points (ppm) Q Q IQ QSIQ. QSJQI 01()+

QB/Q.

Tichodrma 10 2.5 250 0.25 0.13 10.38 2.5 500 0.25 0.25 2-5 1000 0-25 0.5 0.75 2.5 2000 0.25 1 1.25 5 25 0.5 0.01 0.51 50 0.5 0.03 0.53 100to 0.5 0.05 0.55 250 05 0.13 0.68 j 2000 0.5 1 Pscuzdornonas I 500 0.5 0.5 <1I 1000 0.5 1 P M O W I c a ll MPAUM NIMU Mir. O M 16 Example Component A WTC Component B p-hydro~ybenzoic acid Test organism Quantitic; producing end points (ppm) Q Q. QI Q3 QA/Q.. QSiQ., 0'Q-+ 2.5 1000 0.25 0.5 0.75 2.5 2000 0.25 1 1.25 25 0.5 0.01 0.51 L 50 0.5 0.03 0.53 5 250 0.5 0.13 J0-63 5 500 0.5 0.25 J0.75 5 1000 0.5 0.5 1 2000 0.5 1 I_ 100_ 0. 02 0-3 0.25 2.50 0.25 0.5 0.75 0.5 5 0.5 0.01 0.51 0.5 10 0.5 0. 02- 0.52 0.5__0 0.55 IF 100 0.5 0-2 0-7 05 250 0.5 10.5 1__

-[I

IC.

-V.

4-

II~

4 1.7 Ex~mple 6

I

I.

jKK:~ 1:77.

Component A ?-TC Componeat B Dehydroacmic acid Test orgazism Quantitiesproduciog end poimt (ppm) Q. Q Q3 QA/Q- QSIQ' J.

Ticderma 10 2.5 s0 0.25 0.1 0.35 2.5 100 0.25 0.2 0.45 2.5 250 0.25 0.5 0.75 S J- 25 0.5 0.05 0.55 5 50 0.5 0.1 0.6 100 0.5 0.2 0-7 5 1 250 0.5 0.5 1 Fz 500sI- aeuiop 0.25 lw 0.25 10.4 0.65__ 0.5 so 0.5 0.2 0.

100 0.5 0.4 0.

9

I

I'

Example 7 Component A MTC Component B Propionic acid Test organism Quanttes produlcing end Points (jPm) QL Q, Q, QS Q.Q 50 105 0.03 0.53 100 0.5 0.05 0.55 5 250 0.5 0.13 0.63 i 500 0.5 (15 0.75 100 0. 0.5 <1I ~2000 Psed0rgn 1i 100 0-25 04 1- 0.

0. 50 0.5 0.2 0.-7 100 0.5 0O.4 0.9 L L 250 ExamnpeS Component A kfTC Component B =MehYI plr'aben Test organism Quantities podUCin~g end pints IJQ. QSIQ

J.

Tichd 10 I. S00 0.1 0.5 0.6 2.5 s0 0.25 0.05 0.3 2.5 100 0.25 0.1 0.35 2.s 2.50 0.25 0.25 0.

2. 500 0.25 0.5 0.75 5 25 0.5 0.03 0.53 5 50 0.5 0.05 100 0.5 0. 5 250 0.5 0.25 0.75 500 0.5 0.5 <1I 1000 01 1000 0.1 1 1.1 0.25 500 0.25 0.5 0.75 0.25 I1000 0.25 1 1.25 05 50 0.5 0.05 0.55 0.5 100 0.5 0.1 0.6 0.5 250 0.5 0.25 0.75 0.5 500 0.5 0.5 0.5 10 1000- Example 9 Component A =MTC Comiponent B Propyl paraberi Test organism Quantities producing end points 2.5 250 2.5 1005 50 2 .5 250 5 500 -0- 0.01 0.25 0.03 0.25 0.15 0.5 0.25 0. 0.2 QjQ,+

Q./Q

0.6 0.35 0.45 0.75 0.55 0.6 0.7 0.6 0.75 7 0.5 0.51 0.53 0.55 0.6 0.75 21 Example Component A =MTC Component B Nonanoic acd Test organism Quantities producing end points (ppm) Q. QQ. QVQ. JQ.IQ Q. I Q.

Trc o e-a 10 barzianurn_ 2.5 250 0.25 0.5 10.75 250 0.5 ii 500 Pscudprrnnas I 0.25 10 0.25 0.4 0.65 0.5 5 0.5 0.02 0.5 10 0.5 0.04 0.54 I- O 100 t0.5 0.25 20 Li.. e~ 22 Example II Camponent A MTC Component B Otsanoic acid Test organism Quantities producing eod points (ppm) Q_ Q AI QI'Q

Q./Q

Tricohdrima 10 hairum_ so 0.25 0.1 0.35 I 0.25 0.2 0.45 250 0.25 0.5 0.75 25 0.5 0.05 0.55 50 0.5 10.1 0.6 S1 1 100 0.5 0.2 0-7 2-50 0.5 J0.5 1 500- 1 0 2 1 0 0 0 .2 5 0 .4 0 -6 5 0.5 50 0.5 0.2 0.7 J.s IcO 0.5 O 0-9 i .i 3

F;

d

P"

3 Emaiple 12 Cotnneac A NTC ComlaunraL B Uidecylezk acid Tcst organism Quantitits producing end points Q_ H IQ Q. Q.Q T *ghdem 10 0.5 50 0.05 0.25 0.3 1- lo 0.05 0.5 0.55 Soo 0.1 0.25 0.35 I 1000o 0.A 0.5 0.6 2.5 so 0.25 0.03 0.28 2 5 100 0.25 0.05 0.30 I- 25 50 .2 0.25 j 5 0.5 0.01 0.53 50 0. 0-03 0.53 100 0.5 0.05 0.55 S 1250 0.5 0.13 63 I- 0 0.5 0.25 0.75 200 I PdIII- 0.1 250 0.1 0.25 0.35 0. So 0.1 0.5 0.6 0.25 250 0.25 0.25 Ii 0.25 500 0.2.5 0.5 0.75 f 100 0.5 0.1 0.6.

250 0.5 0.25 0.75 r.

r i r a a j fi- 24 Exmple 13 Component A NfTC Component B L-,ric acid Test organism Quantities producing end points (ppm) Q. Q Q. QIIQ QRIQ Trcodrn 10 2.5 Soo 0.2S 0._5 2. 1000 0.25 10.5 0.75 0.5 10. 1 1.25 25 0.5 0.01 0.51 S 50 0.5 M0.03 0.53 5 100 0.5 0.05 0.55 [s j i Jo.lj 0.6s S -0 0.5 10.25 0.75 5 low_ 0.5 0.5 1 s 00 0.5 I1 I >200- 2000 PNudomon w- I 0.25 o o 0.25 10.5 0.75 0.25z 1000 0.25 I 1 1-25 100 U 0.

1 0.6 0.5 2SO 0.5 0.25 0.75 100 Soo 0.5 0 <_I 10o 0-5 1 1000

A

I

25 Example 14 Component A %ITC Component B Formic acid Test organism Quantities producing end points (ppm)_ Q. Q, 2 Q QJQA Q45Q 3

OQ.

2000 0.25 1 1.25 5 250 0.5 0.13 0.63 NO0 0.5 0.25 10.75 000 0.5C 0. <1.I _i 5 2D0O00 5 10. 1.

1- >200 0.25 .50 0.25 0.5 0.75 _0.5 t 5 0 0.5 0.5 I 1- 100

V

-26- Example Component A =MTC Component B =ACiCd a2id Test organism Quantities producing end points (ppm) Q. Q, Q, QJQA Q0I. Q JQ) I 2000 0.1 1 1.1 2000 0.25 1 1.25 500 05S 0.25 0.75 00 0. <1.

2>1 000 0.5 0. <I.

0.25 0. 0.75 0. 50__ O 0.5 0.5 <I -100

IA

-4 27 Example 16 Component A =MTC Compoumern B =Oxalic acid Test organism Quantities producing end points (ppm) 2L QA 2Q Q, QJQA" Q2' m

Q.A'Q.

Tichoderma 10 5 1000 0.5 0.5 <I 2000 0.25 j- 10 0.25 0.1 0.65 0.5 25 0.5 0.1 0.6 0.5 50 0.5 0.2 0.7 It- 0.5 100 -0.5 0. 0.9 250_ -4 28 Example 17 Comnponent A MTC Componeoc B =Citric acid S. S S 4

S.-

A

4 I' 29 Example 19 Component A MTC Component B Malic acid Test organism Quantities producing cod points (ppm) S -a

'A.

U

Iia hau~im

Q.

I0 1QIQA I QSQ. OQ 1000 2000 0.5 0.5 0.5

QSIQI

<I

<1.

>~>2000

I

Pscduron 0.25 100 0.25 0.4 0.65 0.5 10 0.5 0.2 0.9 250 0 L l~ Example 19 Componeat A WMTC Component B Glycolic acid Test organism Quantities producing end points (ppm) Q. Q Q 3 Q.IQA QiQS Q 4

IQ.

TicbodcmL 10 1000 0.5 0.5 I 5 2000 0.5 1 2000 Pseud ompwA 0-25 100 0.25 0.4 0.65 0.5 50 0.5 0.1 0.7 0-5 100 0.5 0.4 0.9 250

T

Claims

2. The composition of claim 1, wherein the microorganism is bacteria or fungi.
3. The composition of claim 2, wherein said bacteria or fungi is Trichoderma harzianum or Pseudomonas aeruginosa.
4. The composition of claim 1, wherein the methylene-bis (thiocyanate) is 2- methylene-bis (thiocyanate). The composition of claim 1, wherein the organic acid is selected from the group consisting of aromatic organic acids, cyclic organic acids, aliphatic organic acids, their respective salts and esters, and mixtures thereof.
6. The composition of claim 5, wherein said organic acid is selected from the group consisting of benzoic acid, sodium benzoate, p-hydroxybenzoic acid, methyl paraben, propyl paraben, dehydroacetic acid, octanoic acid, nonanoic acid, formic acid, sorbic acid, potassium sorbate, acetic acid, oxalic acid, glycolic acid, citric acid, malic acid, propionic acid, lauric acid, undecylenic acid, their respective salts and esters, and mixtures thereof.
7. The composition of claims 1-6, wherein the weight ratio of to is from about 1:30 to about 30:1.
8. The composition of claim 7, wherein the weight ratio of to is from about 1:5 to about 5:1. I: 32
9. The composition of claims 1-6, wherein the weight ratio of concentrations are from about 0.1 to about 500 ppm of methylene-bis (thiocyanate), and from about 0.1 to about 2000 ppm of the organic acid. A method of controlling the growth of at least one microorganism in or on a product, material or medium susceptible to attack by said microorganism comprising the step of applying to said product, material or medium a composition in any one of claims 1-9.
11. The method of claim 10, wherein said product, material or medium is S wood pulp, wood chips, lumber, paints, leather, adhesives coatings, animal hides, tanning liquors, paper mill liquors, metalworking fluids, petrochemicals, pharmaceutical formulations, cooling tower water, cosmetics, toiletry S-formulations, textiles, geological drilling lubricants or agrochemica! compositions for crop or seed protection.
12. The method of claim 10, wherein said components and are applied separately to the product, material or medium, or are first combined and .then added to the product, material or medium.
13. A method for preventing spoilage of a product, material or medium caused by bacteria, fungi or both comprising the step of applying to said product, material or medium, a composition in any one of claims 1-9 in effective amounts to prevent said spoilage. DATED this 22nd day of February, 1999. BUCKMAN LABORATORIES INTERNATIONAL INC WATERMARK PATENT TRADEMARK ATTORNEYS 290 BURWOOD ROAD HAWTHORN VICTORIA 3122 AUSTRALIA IAS/SH DOC 8 AU1525795.WPC I