United States Patent (19) (11) Patent Number: 4,766,113 West et al. 45) Date of Patent: "Aug. 23, 1988

54) ANTMICROBAL COMPOSITIONS AND (521 U.S. Cl...... 514/187; 514/191; METHODS OF USENG SAME 514/576 (75) Inventors: Michael H. West; Fritz J. Nagel, both 58) Field of Search ...... 514/187, 191,576 of Memphis, Tenn. 56 References Cited (73) Assignee: Chapman Chemical Company, U.S. PATENT DOCUMENTS Memphis, Tenn. 2,755,280 7/1956 Feigin et al...... 424/245 X 3,141,821 7/1964 Compeau ...... 424/347 X * Notice: The portion of the term of this patent 3,223,584 12/1965 Luckenbaugh et al...... 424/273 X subsequent to Jul. 22, 2003 has been 4,602,011 7/1986 West et al...... 514/187 disclaimed. OTHER PUBLICATIONS 21 Appl. No.: 854,612 Chemical Abstracts 62:12392b (1965). Chemical Abstracts 72:65702m (1970). 22) Filed: Apr. 22, 1986 Chemical Abstracts 52:12437.h (1958). Primary Examiner-Leonard Schenkman Related U.S. Application Data Attorney, Agent, or Firm-Fred Philpitt 63 Continuation of Ser. No. 419,396, Sep. 17, 1982, Pat. No. 4,602,011, which is a continuation of Ser. No. (57) ABSTRACT 175,073, Aug. 4, 1980, abandoned, which is a continua Our invention pertains to various new compositions, tion-in-part of Ser. No. 002,555, Jan. 11, 1979, aban methods for using such compositions and products doned, which is a continuation of Ser. No. 842,933, Oct. 17, 1977, abandoned, which is a continuation-in treated with such compositions. Our new compositions part of Ser. No. 625,741, Oct. 24, 1975, abandoned, include, among other things, certain antimicrobial which is a continuation-in-part of Ser. No. 364,018, agents solubilized with certain disubstituted aryl com May 25, 1973, abandoned. pounds. 51 Int. Cl.' ...... A01N 43/00; A01N 55/02 19 Claims, No Drawings 4,766,113 1 2 "starved' by chemically disrupting or sequestering the ANTMICROBAL COMPOSITIONS AND excreted enzymes. METHODS OF USING SAME With respect to those antimicrobials that penetrate the semipermeable membranes of the cell, little is This application is a continuation of Ser. No. 419,396, known of the nature of their activity once inside the Sept. 17, 1982 (now U.S. Pat. No. 4,602,011) which in cell. Some researchers believe that they may chelate turn is a continuation of Ser. No. 175,073 Aug. 4, 1980 trace amounts of metals that are necessary to support (now abandoned) which in turn is a continuation-in-part the life of the microorganism, others have suggested of Ser. No. 2,555 Jan. 11, 1979 (now abandoned) which that they bring with then small amounts of metals, such in turn is a continuation of Ser. No. 842,933 Oct. 17, O as mercury, silver and copper, that are toxic to the 1977 (now abandoned) which in turn is a continuation organism, others have suggested that some antimicrobi in-part of Ser. No. 625,741 Oct. 24, 1975 (now aban als may oxidize fatty acids and proteins within the cell, doned) which in turn is a continuation-in-part of Ser. and, in the absence of any positive proof of the precise No. 364,018 May 25, 1973 (now abandoned). The bene activity within the cell, one expert has suggested that fits of 35 USC 120 are claimed relative to all of these 15 when an effective antimicrobial agent enters the highly prior applications. complex and delicately balanced chemical system or the microbe, the result is massive disruption of the system, SECTION I not unlike throwing several cats over a clothesline One embodiment of our invention pertains to an anti while their tails are tied together. microbial agent solubilized with a disubstituted aryl 20 In those instances in which the antimicrobial must compound having a hydrophilic and an oleophilic sub pass through the wall and enter the microorganism cell stituent. In one preferred embodiment the antimicrobial to be effective, it was believed to follow that, since agent is an organometal compound, such as a metal practically all microorganisms live in an aqueous envi complex of 8-hydroxy quinolinol (oxine), and the disub ronment, it should be a criterion that these antimicrobi stituted aryl compound is an alkyl benzene sulfonic 25 als be hydrophilic. While this supposition was correct acid. with respect to penetration of the outermost layers of This invention relates to antimicrobial compositions the microorganisms, it was later recognized that hydro and places particular emphasis on antimicrobial compo philic materials would be repulsed by the lipoid layers sitions that have low toxicity to plant and animal life. of the cell. It was then theorized that an oleophilic The increasing pressures of government and the awak 30 antimicrobial agent should be selected, but, of course, ening concern of the public to environmental protection this was not always effective since the oleophilic (hy are greatly restricting the use of time-honored antimi drophobic) agent could not traverse the outer aqueous crobials for toxicological reasons. For example, in the barrier of the cellular structure. It is now recognized case of fungicides which have been widely used in the that this dilemma may be solved by structuring a com outdoor environments of agriculture and lumber treat 35 posite compound in which one portion of it is hydro ment, many of the known effective fungicides are based philic and another portion of it is oleophilic. upon compounds of toxic metals, such as mercury and It has also been recognized for a number of years that lead, and organic compounds such as chlorinated phe an oleophilic material, in order to penetrate the walls of nols. With increasing population growth, the demands a microorganism, must have some degree of steric com for more and more food production and the increased patibility with the structure of the cell wall. At least one need for wood and its preservation have caused these respected writer, James G. Horsfall, in Principals of classic fungicides to be overworked to the extent that Fungicidal Action (Chronica Botanica Co. 1956, Library observable adverse effects upon man's environment of Congress No. 56-8265), likens the nature of an effec have been felt. tive oleophilic group to a shaped charge such as may be Vast demands exist for compounds to control micro 45 used in armor-piercing explosive shells. Horsfall sug organisms in fields other than agriculture and wood gests that the oleophilic or fat-soluble groups should preservation. These include the treatment of fabrics to more or less match the shape of the fatty groups in the prevent mildew and rot; to inhibit and kill bacterial cell's semipermeable membrane as, by so "shaping the growth; the treatment of surfaces and substrates to ob charge,” it will permit permeation through the critical tain antiseptic conditions for medical, industrial, food 50 fatty barrier of the cell. This may be taken to mean, first processing and household purposes; the formulation of of all, that the fat-soluble group must have a length that ink and paints to prevent mold growth and bacterial is significant with respect to the thickness of the cell decomposition; the prevention and treatment of human wall. Next, the oleophilic group should not be unduly and animal diseases; and on through an almost infinite branched nor contain substituents so large so as to cause spectrum of applications touching our daily lives. 55 steric hindrance to the penetration of the "shaped The disruption of the life cycle of microcellular struc charge' into and through the cell wall. A simple illus tures has received considerable study and, while there tration of an oleophilic "shaped charge' is a straight are still many unproved theories as to their mechanism, chained hydrocarbon having at least about six carbon a number of generalities may be made. For example, it atoms in the chain. has been suggested that in probably a majority of in 60 A classic example of the use of a "shaped charge' to stances, the toxic effect of an antimicrobial agent de penetrate a semipermeable membrane of a microcell lies pends upon its gaining access to the interior of the cell. in the discovery of hexylresorcinol. Here the inventor In somewhat lesser instances, the antimicrobial may be found that the germicidal properties of resorcinol, an effective agent through the functions it performs which were known to be only mediocre, could be sig outside the cell. In this latter regard, such as in the case 65 nificantly improved by substituting a straight-chained of fungi that excrete enzymes to metabolize cellulosic six-carbon alkyl group on the benzene ring of a resorci materials around them and absorb the digested materi nol molecule. The effect was dramatic, at least from the als, it has been proposed that some fungi may be viewpoint of commercial success, as hexylresorcinol a 4,766,113 3 4. generation or so ago became a household word and was been observed that judicious chlorine substitution may a disinfectant that could be found in many a home medi permit the use of shorter alkyl chains. cine cabinet. Throughout the specification and examples, the alkyl To summarize, the prior art recognizes that it is often arylsulfonic acids are referred to in language that might necessary for an effective antimicrobial agent to pass 5 suggest pure substances. For instance, dodecyl benzene through the semipermeable membrane of a microcell if sulfonic acid (DDBSA) is benzene sulfonic acid with a it is to be effective in disrupting the microcellular life. It 12-carbon alkyl chain attached to the benzene ring, but is also known that penetration of the cell may be en the DDBSA actually employed in the examples is a hanced by the use of certain oleophilic "shaped typical sulfonic acid commercially available comprised charges' that have stereochemical compatibility with 10 of a complex alkylbenzene which is a reaction product the structure of the lipoid layer of the cell membrane. It is further known that the ability of the oleophilic of benzene with the tetramer of propylene. Such alkyl "shaped charge' to penetrate a cell wall will be in benzenes are mixtures wherein the average alkyl chain creased if the oleophilic group is associated with a hy length determines the name given to the product. This drophilic group that will serve as its passport through 15 so-called DDBSA, with a typical M.W. of 237 as speci the outer aqueous barrier layer of the cell. It has also fied by one manufacturer, may contain side chain been recognized that many surfactants, due to their lengths, from about C2 to C20, more than one side chain, combined hydrophilic and oleophilic properties, are and side chains in varying positions on the aryl ring. A sometimes effective as antimicrobials to varying de usual specification for DDBSA will call for a minimum grees and in certain environments. 20 of 85% C10 through C12 alkyl side chain lengths. An objective of our invention is to provide an antimi Similarly, while C6 to C24 side chain lengths are crobial composition that is effective in controlling a claimed in this invention, this actually refers to the wide spectrum of microorganisms and which has rela approximate average length, such averages being ob tively low toxicity toward animal and plant life. These tainable by use of alkyl aryl sulfo compounds as re and other objects of this invention are accomplished by 25 ceived from a producer, or by mixtures of two or more formulating a solution comprised of an active antimi compounds, each with a different average alkyl side crobial agent and a disubstituted aryl compound in chain length. which the first substituent is an oleophilic group Pure long-side-chain alkyl aryl sulfonic acids will adapted to penetrate the lipoid layers of microcells and function in this invention, but have no practical value the second substitutent is a hydrophilic group. While it 30 because they are not available commercially and be is believed necessary to form an initial solution of the cause of the extremely high cost. antimicrobial agent with the disubstituted aryl com To determine differences in antifungal efficacy and pound, the antimicrobial agent may, when diluted for physical characteristics of compositions of the inven use, begin to precipitate from solution. This precipita tion prepared with Cu-8-Q and a range of alkyl aryl tion from solution may and usually will become com 35 sulfonic acids, carbon chain lengths of the alkyl group plete after the composition has been applied to a sub attached to the aryl group varying from C8 to C15 were Strate. tested. These included C8 (octylbenzene sulfonic acid), C10 (decyl benzene sulfonic acid), C12 (dodecyl benzene DETAILED DESCRIPTION sulfonic acid-DDBSA), C13 (tridecyl benzene sulfonic The Oleophilic Substituent acid) and C15 (pentadecyl benzene sulfonic acid). Deter The oleophilic substituent of the composition of this mination of antifungal efficiency was made by the test invention should have a degree of stereochemical com method of Example 1 set forth below. patibility with the structure of the semipermeable mem It was determined that optimum antifungal efficacy brane of the cell of a microorganism. As recognized in 45 was obtained in the C10 to C13 range, with C12 the prior art, a common structure meeting this criterion (DDBSA) being the best, by a small margin of little is an essentially unsubstituted straight-chained hydro practical consequence for most applications, all-around carbon having a length that is significant with respect to solubilization of antimicrobial agents such as metal the thickness of a cell wall. This generally requires, as a complexes of oxine. A small but significant reduction in minimum, an alkyl chain with about six carbon atoms in 50 efficacy resulted from the use of C8 and C1s, although it. The alkyl chain, on the other hand, should not be too use of both are within the scope of this invention. long since the mobility of an alkyl chain increases with It also was determined that optimum handling cha increasing chain length and alkyl chains will begin to racteristics-concentrate and use-dilutions stability, coil if they are too long. When the coiling becomes ease of dilution with water, low temperature concen significant, it can cause steric hindrance and make per 55 trate viscosity, maximum amount of antimicrobial meation of the cell wall difficult. While the maximum agents incorporatable in the concentrate, etc.--of com length of the alkyl chain can only be determined with positions of the invention were obtained generally with respect to a given environment and a specific cell struc C12, although differences with C10 and C13 were small ture, it is believed to be a fair generalization to suggest to insignificant in most use instances. that an alkyl chain much longer than 18 carbon atoms C15, because of its greater lipophilic character, exhib and, more especially, one longer than 24 carbon atoms, its diminished water solubility, a matter of practical will lose its effectiveness in penetrating the cell wall. consequence since water is the preferred use diluent for The preferred alkyl groups of this invention are not most end uses of the compositions of this invention. The excessively branched or substituted in the sense that increased lipophilic character imported to the composi they will result in steric hindrance. Nonetheless, some 65 tions by the C15 alkyl chain exerted no adverse effect substitutes along the hydrocarbon chain, such as chlo upon general organic solvent solubility of the invention rine, may improve the oleophilic properties of the compositions and is of benefit in those end uses where, "shaped charge' and may be used to advantage. It has for a variety of reasons, a non-water carrier is desired. 4,766,113 5 6 Lastly, it is within the scope of the invention to em It has been discovered that not only are antimicrobial ploy mixtures of sulfonic acids including alkyl side agents such as metal complexes of oxine readily soluble chains of Co and C1 with C8 to C24. in the substituted aryl compounds of this invention, but No rigid definition can be given as to the length, also their effectiveness as an antimicrobial is materially composition and configuration of the oleophilic group increased over formulations of these antimicrobial and these parameters necessarily must be adjusted for agents known to the prior art. As will be demonstrated the microbial genus and species that are to be con in the following examples, when Cu-8-Q, for example, is trolled. It is believed, however, that given the insight solubilized by dodecyl benzene sulfonic acid (a substi provided by this specification, it will be within the skill tuted aryl compound of this invention), its effectiveness of a worker of ordinary skill in the art to select, with 10 in controlling the growth of microorganisms improves minimum effort, an oleophilic substituent that will per dramatically. meate the lipoid layer of a microcellular organism in the In addition to Cu-8-Q, other metal complexes of practice of this invention. oxine are known to be effective antimicrobials. Of these, mercury is generally regarded as being more effective The Hydrophilic Substituent 15 than copper, and the following is a listing taken from The hydrophilic substituent of the aryl compound literature references of the relative effectiveness of sev should be ionizable and contribute to the solubilization eral metal quinolinolates in descending order of activ of the antimicrobial agent. Two of the most effective ity: mercury, copper, cadmium, nickel, lead, cobalt, and chemically accessable such substituents that will zinc, iron and calcium. Aluminum and tin are also use accomplish this purpose are the sulfo and the hydroxyl 20 ful. radicals. Antimicrobials other than metal oxines may be solu bilized in the alkyl-aryl sulfonic acids of the invention, The Aryl Compound both alone and in the presence of the metal oxines. For A preferred aryl compound is benzene which is be a variety of specialized requirements, such other antimi 25 crobials have significant utility, not only for antimicro lieved to be most effective and, perhaps to a lesser ex bial properties, but also because of the considerable tent, naphthalene. Aryl compounds comprised of more handling and application advantages of true solution than two ring structures may lose their effectiveness for formulations in those instances where the antimicrobial several reasons, among them being the fact that their normally is not water-soluble and must be applied either size becomes large as compared with the oleophilic 30 as a dust or as a particulate suspension in water. Among substituent and will tend sterically to hinder the oleo the antimicrobials which are useful are these classes: philic substituent in penetrating the wall of the microor 1. Oxine and derivatives thereof. ganism. While not essential in the practice of this inven 2. Chlorophenols such as tri-, tetra- and penta-chloro tion, tertiary substituents of the aryl group may some phenol. times prove desirable, particularly if they are capable of 35 3. Anilides such as 2,5-dimethyl-3-furylanilide. withdrawing electrons to improve the solubilization of 4. Benzimidazoles such as 2-(methoxy-carbamoyl)- the antimicrobial agent. Substituents which may func benzimidazole. tion in this manner are, for example, -NO2, -CN and 5. Nitrophenols such as dinitrophenol. -CHO. Electron-withdrawing substituents should be 6. Nitrocresols such as dinitrocresol. used sparingly and with discretion to avoid overloading 7. Crotonates such as 2,4-dinitro-6-(2-octyl)phenyl of the ring structure of the aryl compound. crotonate. The Antimicrobial Agent 8. Organotins such as triphenyltin acetate and tributyl tinoxide. As set forth above, the antimicrobial agent must be 9. Antibiotics such as streptomycin and griseofulvin. able to form a solution with the substituted aryl com 45 10. Organic acids such as acetic, sorbic, salicylic, pound, at least in the initial formation of a concentrate benzoic, dehydroacetic and undecylenic acids. which may later be diluted prior to use. 11. Oxathiius such as 5,6-dihydro-2-methyl-1,4-oxath One particular advantage of this invention lies in the in-3-carboxanilide. fact that antimicrobial agents that have known low 12. Sulfones such as diiodomethyl-p-tolyl sulfone. toxicity toward plant and animal life may be made more 50 13. Iodine, compounds thereof, and iodofors. effective by solubilizing them in accordance with this 14. Aliphatic alcohols from C1 to C18 chain length. invention, thus greatly increasing their utility. Of these, 15. Alkyl, benzyl and phenyl alcohols. for example, the metal complexes (chelates) of 8 16. Hydroxyl diphenyl oxide and sulfide isomers. hydroxy quinolinol (oxine), and especially copper-8- 17. Carbanilides such as trichlorocarbanilide. quinolinolate (Cu-8-Q), are quite prominent due to their 55 18. Bis phenols such as hexachlorophene. comparatively low toxicity toward human, animal and 19. Phenyl mercurials such as phenyl mercuric ace plant life. tate and nitrate. Another unexpected result that accrues through the 20. Complex organic mercurials such as thimersol practice of this invention lies in the fact that when an and nitromersol. aryl-alkyl sulfonic acid is solubilized with a metal-8-Q, 60 21. Aldehydes such as formaldehyde and glutaralde the solution is far less irritating to body tissue and skin hyde. than is the aryl-alkyl sulfonic acid by itself. For exam 22. Aniline derivatives such as 2,6-dichloro-4- ple, in tolerance tests conducted on sensitive areas of nitroaniline. the human skin, solutions of DDBSA and Cu-8-Q di 23. Diphenyl ethers such as 2,4,4'-trichloro-2'- luted 1:200 with water were not only found to be less 65 hydroxyl diphenyl ether. irritating than DDBSA at the same dilution, but also 24. Miscellaneous types including silver oxine; chlor less irritating than cleansing agents generally accepted hexidine and water-soluble acetate and gluconate as being "mild' such as Ivory soap. derivatives thereof; imidazolidinyl urea; hexylre 4,766,113 7 8 sorcinol; 3(2-(3,5-dimethyl-2-oxycyclohexyl)-2- Broadly speaking, for every part by weight of the hydroethyl)glutarimide; benzoaminobenzene sulfo metal-8-quinolinolate, it is preferred to include from 5 nate; 3,ethylidene-L-azetidine-2-carboxylic acid; to 50 parts by weight DDBSA and 1 to 50 parts by 4,6-dichloro-N-(2-chlorophenyl)-1,3,5-triazin-2- weight of the polar diluent. The most preferred compo amine; N,N'-(piperazinediyl bis(2,2,2-trichloroe sition according to the present invention contains from thylidene) bis(formamide); salicyanilide-2- 2 to 10 parts by weight of Cu-8-Q, 25 to 83 parts by hydroxy-N-phenylbenzimide; hexachlorobenzene; weight of the alkylbenzene sulfonic acid, and 15 to 35 boric acid; 5,6,7,8-tetrachloroquinoxaline; 1 parts by weight of the diluent per 100 parts by weight of hydroxypyridine; 2-n-octyl-4-isothiazolin-3-one; the concentrate. One specific composition produced 1,2-benzisothiazolin-3-one; p-chlorophenyl-3-iodo O according to the present invention contains about 5 propargyl formol; dimethylol dimethylhydantoin parts by weight of Cu-8-Q, about 64 parts by weight of 1,3-bis(hydroxymethyl)-5,5-dimethyl-2,4- DDBSA, and about 31 parts by weight of methanol. imidazolidine-dione; 7-(cis-3-chloro-2-propenyl Additionally, it has been found useful to add minor 1,3,5-triazo-7-azoniatricyclo[3.3.1.1.3"decane. amounts, e.g., 5% by weight, of ethylene glycol to In the examples that follow, two methods were used 15 improve shelf life. for incorporating a metal-oxine complex into the com The concentrates prepared as above are diluted with position of the invention: (a) the addition of the complex a carrier, preferably water, prior to use, and essentially itself, and (b) the in situ formation of the complex in the may be diluted to any degree. Other carriers may be compositions. The latter often is preferable for eco used, including xylene, isopropanol, ethylene glycol nomic and versatility reasons. 20 and naphtha. The diluted solutions can be applied by a For in situ formation, an appropriate metal com known technique, such as brushing, spraying, dipping pound is used for reaction with oxine. In the case of or wiping. Cu-8-Q, for example, copper hydrate, copper sulfate, copper acetate, copper chloride and copper naphthen EXAMPLES ate are among the suitable sources of copper. The actual 25 choice depends on the economics (some copper com Examples 1-22 pounds are cheaper copper sources than others), nature In the examples that follow, the effectiveness of the of by-product (copper hydrate and oxine form water; various formulations was determined by treating freshly copper acetate plus oxine form acetic acid, etc.); ease of cut pine boards by dip immersion for 10 seconds in the reaction (some copper compounds release copper to 30 formulation to be tested. The boards, along with an form the copper-oxine chelate easily; others do so more untreated control, were placed in a chamber for the slowly and only under more severe reaction condi period of time indicated and maintained at a tempera tions). All are within the scope of this invention. This is ture of about 80' F. and a humidity of about 70%. true also of in situ formation of metal-oxine chelates At the end of the test period, the boards were re other than copper. 35 moved from the chamber and rated for effectiveness in Generally, oxine itself is the preferred raw material terms of percentage of total surface area covered by for preparation of the metal chelate inasmuch as oxine fungal stain and mold growth. Accordingly, the lower itself is effective, more readily available and lower in the percentage, the higher the effectiveness of the test cost than substituted chelating oxines. The latter, how formulation. ever, are within the scope of this invention and have Unless otherwise indicated, the tabularized formula utility in certain medical applications where more tions were concentrates that were diluted with a carrier, costly substituted oxines is not such a significant limita as indicated, prior to application to the substrate. All tion. parts given in these and other examples are parts by Generally, the full metal chelate of oxine is the pre weight, unless otherwise noted. ferred form, but two variants of full chelates are within 45 the scope of this invention: (a) half-chelates in the case Ex- % Fungal of metals with a valency of two or more, such as cop am- Cu-8-Q Solubilizing Diluent Growth per, and (b) a stoichiometric excess or deficiency of ple Amount Agent (amount (amount) (42 days) metal required to produce the full chelate. 10 DDBSA (50) Methanol (40) 29 A polar diluent is used with the mixture of DDBSA 50 2. 10 98% H2SO4 (20) Water (70) 53 and Cu-8-Q which not only serves as a viscosity-reduc 3 10 Maleic acid (50) Water (40) 58 ing agent, but also permits ionization of the alkylben 4. Untreated control boards 74-90 5 5 DDBSA (64) Methanol (31) 17 zene sulfonic acid to achieve complete solubility of the 6 5 p-toluene sulfonic Methanol (31) 52 Cu-8-Q. It has been found that complete solubilization is acid (64) effected through the use of a highly polar organic sol 55 7 5 1-naphthalene sul- Methanol (31) 74 vent which, for purposes of economy in diluting the fonic acid (64) 8 5 Benzene sulfonic Methanol (31) 92 solution for end use applications, is preferably water acid (64) miscible. A partial list of suitable diluents for use in the 9 5 Methane sulfonic Methanol (31) 62 present invention is given below: acid (64) Methanol 10 Untreated control boards 90 Ethanol Isopropanol (Use dilution in above examples 400:1 with water carrier.) n-Butanol In each of the above examples, the Cu-8-Q formed a Dimethylformamide true solution with the solubilizing agent, both as a con N-methyl-2-pyrrolidone 65 centrate and when diluted for use with the water car Ethylene glycol rier. These examples show that solubilization of the Propylene glycol antimicrobial agent is not sufficient to obtain the desired Water results of this invention, but that the solubilizing agent 4,766,113 10 must be selected in accordance with the criteria dis cussed above, such as is the case with DDBSA which is -continued a disubstituted aryl compound having the defined oleo % Fungal Growth philic and hydrophilic substituents. Example Meta-Oxine (28 days) In the following examples, the very real improved 5 22 Zinc 46 antifungal activity achieved in the practice of this in vention is demonstrated. Cu-8-Q and DDBSA were The foregoing metal-oxines (metal-8-quinolinolates) tested separately and combined to determine efficacy in also may be prepared in situ in the compositions by control of sapstain and mold on fresh cut green lumber. reacting oxine with any of a number of appropriate Also compared is Nylate 10 (Seymour Chemical Com 10 metal compounds. Although the copper chelate of pany), a commercially available, solubilized Cu-8-Q and oxine generally is the most effective and versatile for a water emulsifiable concentrate (10% Cu-8-Q). For use, broad range of end uses, other metal-oxine chelates Nylate 10 was diluted 1:200 with water. have utility. 15 Example 23 Ex am % Fungal Growth The following DDBSA/Cu-8-Q solution was pre ple Composition (28 days) pared in accordance with previously stated techniques. 11 Nylate 10 aqueous emulsion, 0.05% 72 (In this instance, Cu-8-Q was formed in situ from copper Cu-8-Q 20 hydrate and oxine, also known as 8-hydroxy quinoline.) 12 0.025% Cu-8-Qaqueous suspension 89 Copper hydrate 1.70 (prepared by ball milling) Oxine 4.44 13 0.35% DDBSA aqueous solution 49 DDBSA 64.81 14 Prepared by mixing the composition 6 of Examples 12 and 13 to form a solu Methanol. 15.05 tion comprised of 0.025% Cu-8-Q and 25 Isopropanol 14.00 0.35% DDBSA This composition was diluted with a water carrier, as tabularized below, and tested in comparison with a sodium tetrachlorophenate (23%) liquid concentrate, The performance of the composition of the invention also diluted in a water carrier, against organisms on (Example 14) is much superior to that of Cu-8-Q or three species of green lumber-Douglas fir, Amabilis fir DDBSA alone and to that of a conventional solubilized 30 and Ponderosa pine. The organisms were: Cu-8-Q/water emulsion. Cephaloascus fragans a brown mold that infects cer In order to demonstrate the effectiveness of this in tain wood species vention as an antifungal, sapstain and mold control, Trichoderma virgatum a common mold comparisons were made with two standard, widely used Mixed spores a combination of two molds (Penicil antifungal compositions. 35 lium sp. and Aspergillis niger) and a fungus (Cerato cystis pilifera that causes blue stain in wood. % Fungal The freshly cut wood samples were dip treated (15 Growth second immersion) with the test fungicides and then Ex- - (28 days) - innoculated with spore suspensions of the above al- Test Test 40 described fungi. The test boards plus untreated control ple Composition i1 #2 boards were then placed in a warm, humid chamber for 15 Sodium tetrachlorophenate 16.40%. 20 7 four weeks. The results are set forth in the table below Other sodium chlorophenates 4.43% Phenyl mercuric lactate 0.40% in which: Inerts 78.77% 45 (Use dilution - 1:100 in water) 16 Sodium pentachlorophenate 31.6% 20 0 A = Douglas fir O = no growth Other sodium chlorophenols 4.4% B = Amabilis fir 1 = no growth for 2 weeks Borax (sodium tetraborate 57.0% C = Ponderosa pine 2 = medium growth 10 H2O) 3 = heavy growth in 4 weeks Inerts 7.0% 50 4 = heavy growth in 2 weeks (Use dilution - 10 pounds per Use C. T Mixed 100 gallons of water) Dilu- fragans virgatum SPOCS 17 Cu-8-Q 10.0% 17 O tion A B C A B C A B C DDBSA 50.0% Methanol 40.0% Tetrachloro- 1:100 4 4 4 0 O 3 3 4 4 phenol (Use dilution - 1:200 in water carrier) 55 composition DDBSA/Cu-8-Q 1:240 2 0 1 3 0 4 4. 3 4 To demonstrate the antifungal properties of various solution Tetrachloro 1:50 2 0 4 0 0 1 3 2 4 metal-oxine chelates, compositions were prepared using phenol 6 parts by weight of the indicated meta-oxine plus 64 composition parts DDBSA plus 31 parts methanol. These composi- 60 DDBSA/Cu-8-Q 1:120 0 0 1 3 0 1 2 2 2 tions were diluted 1:200 with a water carrier for use. solution Tetrachloro 1:25 2, 2 3 0 0 1 0 0 3 phenol % Fungal Growth composition Example Metal-Oxime (28 days) 65 DDBSA/Cu-8-Q 1:60 0 0 0 1 0 O O 0 0 solution 8 Copper 17 Tetrachloro 1:12.5 0 1 0. O 0 0 O 0 2 19 Tin 20 phenol 20 Aluminum 28 composition 2 Nickel 39 DDBSA/Cu-8-Q 1:30 0 0 O O 0 0 O 0 O 4,766,113 11 12 -continued the following table. Minimum fungicidal concentrations solution were determined against a broad spectrum of fungi that Control (no 4 4 4 3 4 4 4 4 4 are detrimental to man, foodstuffs and materials and treatment) which can result in metabolite formations (mycotoxins) of extreme toxicity to man and animals. In the table below, Composition A is that of this Example 24 Example 25 and the active ingredient is Cu-8-Q;0; Com To further illustrate the antifungal properties of com position B is DDBSA; Composition C is pentachloro positions of the invention, the formulation below was phenol; and Composition D is 2,3,5-trichloro-4-propyl prepared and tested as a preservative against microbio 10 sulfonyl pyridine. Where no concentration is listed, no logical deterioration of 10 oz. cotton duck cloth and data is available. The stated value of '1' means 1 or compared with untreated cotton duck as a control and fewer ppm. with Cunilate 2174, a commercially available concen trate containing 10% Cu-8-Q which is made soluble in 15 Minimum Fungicidal Concentration petroleum hydrocarbon solvents via use of nickel ace - (ppm of active ingredient) - tate and 2-ethylhexoic acid. The previously described A B C D Nylate 10 also was tested. Aspergilis niger 1. 10,000 1-3 - Oxine 4.1 parts by weight (ATCC 9642) Copper hydrate 1.4 parts by weight Aspergilis terreus 1 100 - 36 DDBSA 64.0 parts by weight 20 (ATCC 10609) Aspergillis flavus 1 1,000 22-54 - Propylene glycol methyl ether 30.5 parts by weight (ATCC 11655) This composition was use-diluted 1:24 with a water Alternaria alternata l - - - carrier; the Cunilate 2174 was diluted 1:19 and 1:9 with (ATCC 13963) mineral spirits for use; the Nylate 10 was diluted 1:19 Aureobasidium pullulans l 100 - - 25 (ATCC 16624) and 1:9 with water. The cotton samples were dipped to Lenzites trabea 1 100 1-3 - refusal in the test compositions, air-dried and buried at (ATCC 11539) 75' F. for 29 days in sheep manure moistened with Polyporus tulipiferae l 100 1-3 - water. Microorganism attack on the cotton cloth in this (ATCC 11245) Penicilium brevi 1 100 - - test medium is both rapid and severe as can be noted 30 compactum (ATCC 16024) from the essentially total destruction of the untreated Rhizopus stolonifer 1 100 1-3 --- control cloth sample. The results of this test are tabu (ATCC 24794) lated as follows: Trichoderna viride 0 100 - - (ATCC 8678) Trichoderma sp. 100 - u Weight of Cu-8-Q Estimated 35 (ATCC 12668) Composition (dilution) in Cloth Strength Loss' Candida albicans 1,000 - 3 (ATCC 10259) Untreated control O 100% Cunilate 2174 (1:19) 0.41 gram 50% Cunilate 2174 (1:9) 0.84 gram 25% Nylate 10 (1:19) 0.61 gram 50% These results illustrate the high efficacy of the com Nylate 10 (1:9) 1.21 gram 0% position of this invention and confirm the fact that an DDBSA/Cu-8-Q (1:24) 0.18 gram 0% antifungal composition prepared from a Cu-8- "As measured by tear strength reduction: Q/DDBSA solution is much superior to DDBSA alone. 100% = total loss of strength 50% = moderately difficult to tear by hand The results also indicate the favorable relative efficacy 25% = difficult to tear by hand of the test composition compared to the two commer 0% = impossible to tear by hand 45 cially available fungicides of recognized high perfor aCe The superiority of the formulation of the invention over other Cu-8-Q compositions is clearly evident. Example 26 Example 25 Test compositions were evaluated as wood preserva 50 tives via a standard soil block culture procedure The following composition was tested for minimum (ASTM D4131-61) wherein the wood blocks were wa fungicidal concentration and compared to two well ter-leached in accordance with standard technique known antifungal chemicals-pentachlorophenol and prior to exposure to the test fungus. The test fungi were 2,3,5 trichloro-4-propyl-sulfonyl pyridine-and those specified for wood decay evaluation by the Amer DDBSA 55 ican Wood Preservers' Association (AWPA)-namely, Oxine 8.2 parts by weight Lenzites trabea (Madison 617, ATCC 11539) which is Copper hydrate 2.8 parts by weight the standard test fungus for above-ground wood expo DDBSA 59.0 parts by weight sure, and Poria monticola (Madison 698, ATCC 11538) Propylene glycol methyl ether 30.0 parts by weight which is the standard copper-tolerant fungus for ground All of the test composition concentrations to be 60 contact wood use. tested were incorporated in the fungal growth media In the table below, the results are expressed as per (agar) in accordance with standard microbiological centage weight loss of the wood test blocks from decay practices. Agar plugs containing the test fungicides then fungi attack. were inoculated with a sporulating culture and inocu Composition A is comprised of: lated at the temperatures and times specified by The 65 Oxine 4.42 parts by weight American Type Culture Collection (ATCC) recom Copper hydrate 1.51 parts by weight mendations. The plugs were then scored for absence or DDBSA 64.07 parts by weight presence of organism growth. The results are shown in Methanol 30.00 parts by weight 4,766,113 13 14 The composition was diluted 1:110 in a water carrier for 1 = traces of growth impregnation of the L. trabea test blocks and 1:55 for 2=slight growth test against P. monticola, 3= moderate growth Composition B was the same as Composition A ex 4= covered with fungi cept that it was diluted 1:55 with toluene carrier for block impregnation for test against both test fungi. Composition C was the same as the DDBSA/Cu-8-Q Test Composition solution of Example 25 diluted with a water carrier A B C D E 1:220 for test against L. trabea and 1:55 against P. mon Blue stain fungi growth 0.8 0.3 0.5 2.5 3.9 ticola. 10 Mold fungi growth 0.5 0 0.3 1.3 3.6 Cunilate 2174 was diluted 1:110 with a toluene car 1. The demonstrated efficacy of Compositions A and B The abbreviation "pcf” means pounds of Cu-8-Q per against the six listed fungal organisms has utility not cubic foot of wood. only on the tested substrate-wood-but also for pro 15 tection of a variety of other materials that are attacked by one or more of the fungi, including paint, concrete, Lenzites trabea Poria nonticola brick, textiles and leather. Retention Weight Retention Weight Composition (pcf) Loss (%) (pcf) Loss (%) Example 28 A 0.021 0.3 0.040 1.3 20 Using the standard AOAC fungicidal test method B 0.025 5.0 0.029 11.3 C 0.018 2.0 0.036 8.1 (12th Edition, 1975), the composition below was evalu Cunilate 2174 0.024 13.6 0.026 43.4 ated against two widespread fungi. Untreated control O 45.3 O 55.1 Oxine 2.08 parts by weight Copper hydrate 0.70 parts by weight 25 DDBSA 40.00 parts by weight These results demonstrate the efficacy of the compo Isopropanol 32.00 parts by weight sitions of the invention in both a water and an organic Demineralized water 25.22 parts by weight solvent (toluene) carrier. Also illustrated is the greatly The two fungi were Aspergillis niger, a ubiquitous improved efficacy over Cunilate 2174, especially for black fungus which flourishes on a broad range of sub wood in ground contact service. The wood protection 30 strates, and Trichophyton mentagrophytes, a cause of results with Compositions A, B and C compare favor "athlete's foot.' ably with those of pentachlorophenol (PCP), tested A. niger-at 1:200 use dilution in a water carrier, no simultaneously, wherein PCP, a world standard for growth after 10 minutes' exposure. wood preservation, exhibited 1.4% weight loss at 0.30 T. mentagrophytes-at 1:750 use dilution in a water pcfretention against L. trabea and 3.1% weight loss at 35 carrier, no growth after 10 minutes' exposure. 0.27 pcf retention in the wood against P. moniticola. Similar but somewhat lower efficacy results were Employing AWPA Test Method M12-72 (revised obtained by substituting zinc-8-Q or aluminum-8-Q in 1973) for testing wood block resistance to termite the composition of this example, produced by reacting (Reticulitermes flavipes) attack, it was determined that zinc oxide and aluminum hydroxide respectively with no attack occurred at retentions of Composition A ade quate to protect the wood from decay. OXC. Example 29 Example 27 The compositions of this invention exhibit efficacy The purpose of this test was to determine antifungal against a broad spectrum fungal plant pathogens, as efficacy of DDBSA/Cu-8-Q solutions and to compare 45 illustrated by various use dilutions in a water carrier of their efficacy to that of a world standard, sodium penta the following composition: chlorophenate, and a mixture of two well-known agri Copper hydrate 1.70 parts by weight cultural fungicides, Topsin M (a thiophenate) and 8-hydroxy quinoline 4.44 parts by weight Nabam (a thiocarbomate). The test method is desig Isopropanol 35.00 parts by weight nated as a proposal for the Finnish NWPC Standard 50 DDBSA 58.86 parts by weight No. 1.4.1.3/1974. The test substrate was fresh cut, green A. Valencia Oranges pine wood. The test fungi were: Tested on harvested fruit against Phomopsis stemend Blue stain fungi-mixture of Ceratocystis pilifera ZII rot and Diplodis rot, at a 1:100 use dilution, 2-minute Sclerophoma entoxylina Z17 Pullularia pullulans U2 dip application. After 3 weeks at 70' F., the following Mold fungi-mixture of Paecilomyces varioti X15 55 percentages of decay were noted: Cladosporium sphaerospermum R7 Aspergilis am Control (untreated) oranges-9.5% decay stelodami X19 Treated oranges-5.3% decay In the table below, Composition A is the formulation B. Sugar Cane of Example 5 diluted 1:200 with a water carrier. An agar seeding test against Ceratocystis paradoxa Composition B is the formulation of Example 5 di (pineapple disease) at a 1:10,000 (100 ppm) use dilution luted 1:100 with a water carrier. yielded a 3.0 mm. inhibition zone. Composition C is a 1.5% concentration of sodium C. Peach Trees pentachlorophenate in water. Tested against Taphrina deformans (causes leaf curl Composition D is a 0.4% concentration in water of a disease). Four test trees were sprayed twice, two weeks 47:53 weight ratio of Topsin M:Nabam. 65 apart, with a 1:400 use dilution. Three months later, 100 E refers to untreated control pine boards. leaves on each test tree were rated for leaf curl: The rating index is: Control (untreated) leaves-100% leaf curl 0=no visible growth Treated leaves-13.5% leaf curl 4,766,113 15 16 D. Cotton crotonate), Vitavax (5,6-dihydro-2-methyl-1,4-oxathin Effectiveness against 11 fungi and 1 bacterium (Xan 3-carboxanilide), Daconil (tetrachloroisophthalonitrile) thomonas malvacearum) that are associated with disease and Maneb (manganese ethylenebisdithiocarbamate). of cottonseed, seedlings and other plants was evaluated The plants and diseases tested were bean mildew in vitro, using the following compositions: 5 (Erysiphe polygoni), bean rust (Uromyces phaseoli typica) and rice spot (Helminthosporium orazae and Cerocospora Composition #1 Copper hydrate 1.70 parts by weight 8-hydroxy quinoline 4.44 parts by weight orazae). Methanol 4.00 parts by weight Isopropanol 30.86 parts by weight 10 DDBSA 59.00 parts by weight Bean Mildew Bean Rust Rice Spot Composition #2 Copper hydrate 2.80 parts by weight 8-hydroxyquinoline 8.20 parts by weight Example 28 composition 8 10 O Methanol 4.00 parts by weight Karathane 10 P - Isopropanol 26.00 parts by weight Vitavax - 9 MO DDBSA 59.00 parts by weight Daconil -- - 10 15 Maneb --- 8 Both compositions were prepared in accordance with procedures stated in previous examples. Example 31 The following results were obtained, expressed in parts per million (ppm) of total test composition in 20 The following composition was evaluated (diluted water carrier and the relative growth inhibition pro with water for use) in vitro for inhibition against two vided at each test strength on each tested organism. In fungal pathogens, Botrytis sp. and Alternaria sp., causal the tables below: agents of a variety of plant diseases. 0=no apparent inhibition Oxine 2.08 parts by weight l=some inhibition 25 Copper hydrate 0.70 parts by weight 2= considerable inhibition (little growth) DDBSA 40.00 parts by weight 3=total inhibition (no growth) Triton X-100 2000 parts by weight Isopropanol 22.00 parts by weight Concentrations (ppm) Water (demineralized) 15.22 parts by weight Test Organisms 0 1 S. 25 100 500 1000 30 The zone of inhibition agar plate test also was used to Composition #1 test Cunilate 2174 (diluted in mineral spirits for use) for Pythium ultimum (41B) 0 0 0 1 2 3 3 comparison. The composition concentrations in the Rhizoctonia solani (1D) 0 0 0 1 2 3 3 Fusarium (4A) 0 0 0 3 3 3 3 table of results below are expressed in parts per million Fusarium (4D) 0 2 2 2 3 35 (ppm) of Cu-8-Q. The larger the inhibition zone, the Fusarium roseum (4C) 0 0 0 1 2 2 3 greater is the efficacy of the composition. Colletotrichum 0 0 0 2 2. 2 gossypii (35A) Xanthononas 0 0 0 0 2 2 3 malvacearum (2A) Botrytis Alternaria Composition #2 DDBSA/Cu-8-Q Pythium ultimum (41B) 0 0 1 2 2 2 2 Concentration (ppm) 2 50 125 2 50 125 Rhizoctonia solani (1D) 0 0 1 2 2 2 2 Fusarium (4A) 0 0 1 1 1 2 2 Inhibition zone (mm) 13 15 22 O 16 21 Fusarium (4D) 0 0 1 1 2 2 2 Cunilate 274 Fusarium roseum (4C) 0 0 0 2 2 2 2 Concentration (ppm) 83 200 500 83 200 500 Colletotrichum 0 0 0 2 2 2 2 gossypii (35A) 45 Inhibition zone (mm) 3 14 16 0 0 14 Xanthomonas 0 0 0 1 2 2 malvacearum (2A) The DDBSA/Cu-8-Q solution of this invention ex Concentrations (ppm) hibits an improvement in efficacy against the tested Test Organisms 500 1000 organisms by a factor of 4x in the case of Botrytis to 10x Composition #1 50 in the case of Alternaria. Aspergillis sp. Helminthosporium oryzae Example 32 Mucor mucedo Penicillium sp. The DDBSA/Cu-8-Q solution of the preceding ex Rhizopus sp. 55 ample (31), diluted 1:400 in a water carrier, was applied by spray nine times, at two-week intervals, to peach and nectarine cultivars during the growing season. The Example 30 results against brown rot (Monolinia fructocola), com The composition of Example 28 was screened for pared to nontreated trees, is presented below. fungal pathogen response as a foliar spray on beans and 60 rice. The rating scale is from 0 (no pathogen control) to 10 (complete pathogen control). The concentration of % Fruit Affected active ingredient (in a water carrier) of all compositions Peach Nectarine tested is 33 parts per million (ppm). The active ingredi At harvest (treated) 1 3 ent in the composition of Example 28 is expressed in 65 At harvest (untreated) 15 43 terms of Cu-8-Q and the chemical as listed below for Five days later (treated) 3 6 four comparative products. The comparative products Five days later (untreated) 60 67 tested were Karathane (2,4-dinitro-6-(2-octyl phenyl 4,766,113 17 18 which are difficult to impossible to control with pres Example 33 ently available antibiotics, notably Pseudomonas sp. The DDBSA/Cu-8-Q solution of Example 31 was typified by Pseudomonas aeruginosa PRD-10, the stan tested in vitro against a major turf pathogen, Helmin dard strain in the United States for evaluation of anti thosporium vagans, via a standard agar plate culture 5 bacterials for mandatory Gram-negative control appli technique, with these results: cations. The compositions herein disclosed eliminate or sub stantially reduce these use limitations inherent in many Fungus Colony Diameter other germicides. The compositions are quite unique in 1:6700 use dilution 1 mm. 10 in water carrier having strong Gram-positive and Gram-negative activ 1:3350 use dilution O ity combined with broad versatility of formulation and in water carrier use plus a high degree of safety (low toxicity and zero to Control 21 mm low skin and eye irritation). Add to this the high effi cacy, broad spectrum antifungal activity of the compo The results demonstrate very high efficacy in con 15 sitions of this invention and the resulting range of toxic trolling this important pathogen. Complete control of ity to target organisms and safety to man, the most H. vagans was achieved in this assay between 3.7 and 7.5 sensitive of hosts, is unique indeed. ppm of Cu-8-Q. The balance of toxicity provided by this invention to As illustrated in the examples to follow, the composi Gram-positive, Gram-negative and fungal microorgan tions of this invention have high efficacy against a broad 20 isms has special value in the broad consumer field of spectrum of bacteria that are pathogenic to mammals skin deodorancy. Present antibacterials suffer from the and plant life, that contribute to reduced water quality, fact that they are effective primarily against Gram-posi that cause deterioration of foodstuffs, that degrade a tives, allowing Gram-negative and fungi overgrowth, a broad range of manufactured and natural materials and condition considered dangerous by many authorities. products, and which generate toxic metabolites (bac 25 Example 34 teria-toxins) that are among the most poisonous sub stances known to man. The DDBSA/Cu-8-Q solution of Example 25 was Of particular interest is high efficacy against Gram tested, along with a number of well known antimicro negative as well as Gram-positive microorganisms. Few bial agents, against abroad spectrum screen of economi antibacterial materials now available are effective 30 cally important Gram-positive and Gram-negative bac against the Gram-negatives and still fewer provide eco teria. All antimicrobial agents were incorporated in the nomical control of them. A number of available antibac agar bacterial growth media according to standard mi terials toxic to Gram-negative organisms have practical crobiological practices. The bacterial species were limitations which severely restrict use, including high grown in nutrient broth; 24-hour cultures, the inocu mammalian toxicity, phytotoxicity, corrosiveness to 35 lum, then were streaked onto the nutrient agar plates skin and a variety of materials, strong odor, strong containing the test antimicrobials. After a 24-hour incu color, high volatility, low or erratic shelf stability, low bation at the appropriate temperature, the plates were or nonexistent residual activity, and prohibition of use rated for presence or absence of bacterial growth. at elevated temperatures. Minimum bactericidal concentrations for each of the The basic significance in the need for Gram-negative tested agents are stated in the following tabulation of control lies in the fact that this bacterial category in results in parts per million (ppm) of active ingredient as cludes a number of widespread, virulent pathogens defined in the description of each agent.

Antimicrobial Agent (ppm of active ingredient) Bacteria (ATCC No.) A B C D E F G

Bacillus cereus 100 - 7 8 5-10 -- Bacillus lichenforms (27326) 1. 100 - 7 8 2-5 -- Bacillus megaterium (27327) 100 7 8 - - Bacillus subtilis (37328) 100 - 750 8 - 3 Micrococcus flavus (10240) 100 - 7 8 - - Mycobacteriumphlei (15610) 10 --- 7 8 - 3 Staphylococcus aureus (6538) 1 OO 2083 7 8 1-3 3 Gram-negative: Alcaligenes faecalis (337) 10 1000 - 750 80 - - Alcaligenes marshali (21030) 104 100 H 7 8 - - Esherichia coli (11229) 04 10,000 - 750 80 250-500 165 Flavobacterium arboresceus (4358) 10 10 4166 7 8 - - Klebsiella pneumoniae (4356) 10 10,000 - 750 - - Proteus vulgaris 10 1000 - 750 800 -- - Pseudomonas aeruginosa (15442) 104 1000 4166 750 800 1000-2500 65 Salmonella cholerasuis (10708) 04 1000 -- 750 80 250-500 65 4,766,113 19 20 -continued Antimicrobial Agent (ppm" of active ingredient) - Bacteria (ATCC No.) A B C D E F. G Salmonella typhi (6539) 104 1000 2083 750 80 - 165 'stated value of "1" means l or less A = the DDBSA/Cu-8-Q solution of Example 25 with the active ingredient expressed in terms of Cu-8-Q. B - DOBSA. C = phenol D = Betadine, an iodine/polyvinylpyrrolidone complex containing 0.75% iodine. The active ingredient is iodine. E = Alkyl dimethyl ammonium chlorides (61% C2, 23% C, li?a C16 and 3%. Co). F = sodium pentachlorophenate G = 2,3,5-trichloro-4-propylsulfonyl pyridine. These data demonstrate the high efficacy of the com- 15 position of Example 25. On the basis of the average of Example 36 the efficacies againstall the test organisms, Composition This composition was prepared and tested at one use A is 45 times superior to Composition B; 88 times better dilution, 1:50 in water carrier, against the causal agent than Composition C; 10.7 times better than Composition of potato ring rot bacteria (Corynebacterium sepedoni D; and 3.5 times superior to Composition E. 20 cum): On the basis of the average of the efficacies against Oxine 2.08 parts by weight the three test bacteria (Staphylococcus aureus, Salmo- Copper hydrate 0.70 parts by weight nella cholerasuis and Pseudomonas aeruginosa PRD-10) DDBSA 40.00 parts by weight required by the Environmental Protection Agency of a Triton X-100 20.00 parts by weight "hospital grade" disinfectant, Composition A is 10 times 25 Isopropanol 22.00 parts by weight better than Composition B; 7.2 times better than Com- Water (demineralized) 15.22 parts by weight position D; and 1.6 times better than Composition E. Infected potato seed readily contaminate potato seed cutters, sacks, bins, cellars, trucks and planting equip Example 35 ment with the highly infectious ring rot bacteria. The The composition set forth below was prepared by 30 result may be infected potato plants, tubers and reduced previously described procedures: yields. Oxine 2.08 parts by weight The test procedure consisted of dipping unpainted, Copper hydrate 0.70 parts by weight planed wood laths (6') into a slurry of infected ring rot Isopropanol 32.00 parts by weight tuber tissue, allowing excess slurry to drain off (3-5 DDBSA 40.00 parts by weight 35 minutes) and then spraying the contaminated lath with Water (demineralized) 25.22 parts by weight the test antibacterials. Three to five minutes later, When evaluated as a bactericide by the AOAC Use healthy Norgold Russet potato seed pieces were rubbed Dilution Method (12th Edition, 1975), 10 ring carriers vigorously against both sides of the contaminated and per organism, the following results were obtained (A=- antibacterial-treated laths. The process was repeated subculture and B = resubculture): 40 using laths not contaminated with C. Sepedonicum but treated with the test antibacterial agent. The rubbed Pio Negative - Positive seedappropriate pieces weretime. stored in bags and later planted at the in Water Carrier A B A B In addition to the composition of the invention, un Staphylococcus aureus 1:1000 19 19 9 9 45 treated controls, 20% Clorox (1.05% sodium hypochlo igi cholerasuis 1:1000 10 10 0 0 rite in water), formaldehyde (37% formalin diluted Pseudomonas aeruginosa 1:400 0 10 0 0 1:120 in water) and Roccal (benzalkonium chloride or Aerobacter aerogenes 1:400 10 10 0 0 sephiran chloride) diluted with water to 800 ppm con centration were tested. The results of the test are tabu A ten-minute kill is required against the first three 50 lated below and refer to plants and tubers produced pathogens for sale as a hospital grade disinfectant. Effi from the tubbed test seed pieces.

Ring Rot % Plant % Ring Rot % Ring Rot Yield Antimicrobial Contaminated Stored Plants Tubers cwtacre None (control) Yes 98 23 8 493 None (control) No 95 O O 609 DDBSA/Cu-8-Q Yes 98 0 2 631 DDBSA/Cu-8-Q No 100 O O 602 20% Clorox Yes 98 20 9 S00 20% Clorox No 98 O O 602 Roccal Yes 98 20 9 55 Roccal No 98 O O 638 cacy against the fourth organism, a major cause of slime 65 The composition of this example demonstrates supe in recirculated cooling water systems and pulp and rior control of the ring rot bacterium. Other species of paper mills, demonstrates utility of the composition as a the genus Corynebacterium are causal agents of disease slimicide. in man and a variety of plant life. 4,766,113 21 22 Example 37 Th e composition below was prepared and tested for UnmedicatedAverage No. Soap of TestAverage Composition No. of %. Re speed and range of antibacterial activity, in the absence Subject Basin Organisms/Basin Organisms/Basin duction and presence of organic matter (blood) for use in hospi- 5 2,500,000 60,000 97.6 tal disinfection, cold sterilization and antisepsis. 4 500,000 25,000 98.7 Oxine 4.1 parts by weight 5 1,300,000 15,000 98.8 55ESAg 14 parts by weight 2 : 38: : 2: .0 parts by weight 5 800,000 1,000 99.9 Propylene glycol methyl ether 29.5 parts by weight 10 3 l 800,000 40,000 95.0 Many antimicrobial agents are partially or totally 4. 600,000 20,000 96.7 deactivated in the presence of organic matter, constitut- 5 500,000 1,000 99.8 ing a severelimitation to effectiveness of such agents for 4 } : s: 3. a number of uses such as wound antisepsis and medical 5 800,000 loo 95.9 instrumentation and surface disinfection where large 15 5 1. 1,500,000 40,000 97.3 amounts of organic matter often are encountered and 4. 800,000 1,000 99.9 sometimes- are unavoidable. 6 5l 1900,000500,000 50,0001,000 97.499.8 The AOAC Use Dilution Confirmation Test (12th 4 sooooo 15,000 98.1 Edition, 1970) was modified as follows: 5 520,000 1,000 99.8 (a) The test temperature was 37 C. 20 (b) The ring carriers were soaked in sheep blood for two hours, air-dried for one hour, then contami- Example 39 nated with the test pathogen. Oxine 2.08 (c) The contaminated rings were contacted with the Copper hydrate 0.70 test antibacterial agent for 30-second, one-minute 25 Nonyl phenol/EO surfactant* 20.00 and three-minute periods. DDBSA 45.00 The results are set forth in the following table, in Isopropanol 15.00 which: Water (distilled). 17.22 0=no growth in 10 of 10 tubes tested (1 mol nonyl phenol to 6 mols ethylene oxide) 1=growth in 1 of 10 tubes tested 30 2=growth in 2 of 10 tubes tested The above composition was prepared by the proce 3=growth in 3 of 10 tubes tested etc. dure already described and evaluated for efficacy

Use Oilution in In Absence of Blood In Presence of Blood Test Pathogen Water Carrier 30 sec. 1 min. 3 min. 30 sec. 1 min. 3 min. Staphylococcus aureus Undiluted 0 O 0 O O O (ATCC 6538) :0 0 O O O 0 O 1:100 0 O O 3 6 3 Salmonella typhi Undiluted 0 0 0 0 O 0 (ATCC 6539) 1:10 O O 0 0 O 0 1:00 0 O O O 0 O Pseudomonas aeruginosa Undiluted O O O O O O (ATCC 15442) 1:10 0 O 0 10 10 O 1:100 O O O 10 3 3

These results indicate that the test composition is against ciliated protozoan and two types of viruses. capable of rapid antibacterial action in the presence of substantial amounts of organic matter against the three Hemaglutination Assays human pathogens generally considered as definitive for HA Titre 50 antibacterial efficacy evaluation. Virus Untreated Treated (1:50)* Treated (1:200) Example 38 Adenovirus 128 O O Newcastle Dis- 512 O O The composition of Example 28 was prepared for ease virus evaluation as a skin degerming agent against resident (1:50 and 1:200 use dilutions of composition in a water carrier in contact with virus and transient flora. Six subjects were tested using the 55 suspensions for 15 minutes) Modified Prices Multiple Basin Technique which mea sures reduction of skin flora as a percentage of that The HA titre is a measure of the number of infectious achieved by washing the hands with unmedicated soap. virus particles present in the test suspension. Bacterial counts were taken from the first, fourth and fifth basins. The counts from the first basin represent the 60 Plaque Assays transient bacterial flora and that of the fourth and fifth Via the same procedure as above, the untreated virus basins the resident flora. Prior to the test, none of the suspensions contained 6.4x 10 pfu/ml of Adenovirus subjects used a medicated soap for one week. virus particles and 21 x 10 pfu/ml Newcastle Disease The results set forth in the following table demon virus particles respectively. After treatment with the strate a very high order of efficacy for the test composi 65 1:50 and 1:200 use dilutions, readings of 0 pfu/ml were tion for critical degerming uses such as a surgical scrub obtained. Each pfu represents one infectious virus parti for operating room personnel and skin preparation at cle. A zero pfu reading represents total inactivation of the surgical site. the infectious virus. 4,766,113 23 24 (d) A 5.0% pentachlorophenol solution in mineral Protozoan inhibition spirits containing 4% propylene glycol ether to Inhibition of growth of ciliated protozoan (Tetrahy provide sufficient pentachlorophenol solubility and mena) in pond water was obtained at a 6 ppm concen to prevent sublimation from the wood. tration (based on Cu-8-Q) of the above test composition Dry southern yellow pine stakes, ' squarex 24' of DDBSA and Cu-8-Q after 6-hour and 72-hour con long, were dip-impregnated with the test solutions, tacttimes. The 6 ppm reading represents the MIC (Mini allowed to dry for one week, and then buried to a depth mum Inhibitory Concentration). These results demon of 12' in the ground for 15 months, at which time these strate high efficacy of the test composition against the results were observed: test microorganisms. 10 Example 40 Solution Pickup Termite In some instances, it may prove advantageous to Treatment (bs/ft) Rating" include other microbicides in a given formulation to None (control stakes) 36 increase the over-all spectrum of antimicrobial activity. 15 Example 12 1.79 83 Such additional microbicides need not necessarily be of Example 18 1.13 80 5% pentachlorophenol 1.28 84 the type thus far described in this specification. For solution example, when treating green lumber to inhibit sapstain Cunilate 2174 1.26 56 or mold, it is sometimes desirable, to get broader protec (Termiterating: tion, to include a chlorophenol such as pentachlorophe 20 O = stakes totally destroyed nol, tetrachlorophenol, or 2,4,5-trichlorophenol, in the 100 = stakes unattacked) composition of this invention. Also, to provide addi tionally against insect attack, it is within the scope of Both compositions of this invention exhibited termite this invention to include insecticides such as Lindane or control essentially equal to that of the pentachlorophe DDT. 25 nol solution and definitely superior to both the un Further, it has been found that boric acid can be treated control stakes and those treated with Cunilate added to the class of compositions of this invention with 2174. beneficial microbiocidal results: Copper hydrate 0035 Examples 42 to 50 8-hydroxyquinoline 0,104 30 AS further evidence of the effectiveness of antimicro Methanol 0736 DDBSA 1,630 bials prepared in accordance with this invention, anum. Orthoboric acid 2.500 ber of tests were conducted utilizing DDBSA as the Water 94.995 disubstituted aryl compound of this invention as a solu The above concentrate, prepared as previously de 35 tion with the below-listed microbicidal agents. A 1:200 scribed, was diluted 1:9 with water and fresh, rough use dilution in a water carrier was prepared from con sawn, green southern yellow pine lumber was treated centrates containing 65 parts DDBSA, 30 parts metha by 10-second dip immersion. Sample boards were nol, and 5 parts of the antimicrobial agent. Fresh-cut placed, together with an equal number of untreated green pine test boards were dip-treated and evaluated control boards and other test specimens in a chamber 40 for stain and mold after 30 days in the aforementioned maintained at 75 F. and 70% relative humidity for 28 constant atmosphere test chamber: days. %. Moid Example Antimicrobial Agent and Stain % Stain and Mold 45 42 Cu-8-Q 3 DDBSA/Cu-8-Q composition of Example 5 6 43 2-(4-thiazolyl)benzimidazole 14 1:200 use dilution in water 44 cis-N-(trichloromethyl)thio- 23 DDBSA/Cu-8-Q/orthoboric acid solution 7 described immediately above 4-cyclohexane-1,2-dicarboximide DDBSA/Cu-8-Q composition of Example 5 14 45 diiodomethyl-para-tolyl sulfone 8 1:400 use dilution in water 50 46 para-chlorophenol diiodomethyl sulfone 6 Orthoboric acid, 0.50% in water 62 47 2-n-octyl-4-isothiazolin-3-one O Orthoboric acid, 0.25% in water 86 48 2-benzisothiazolin-3-one 14 Untreated control boards 88 49 Mergal BCM 2 50 8-hydroxyquinoline 10 Example 41 55 Example 51 Some of the compositions of this invention exhibit unexpected insecticidal activity when compared to the The following composition was prepared and evalu conventional solubilized Cu-8-Q compositions and ated for control of pineapple disease of sugar cane when compared to a known insecticide (pentachloro (Ceratocystis pilifera): phenol). Against termites in a soil burial test in Mem 2-(methoxy-carbamoyl)-benzinidazole 4.0 phis, Tenn., the following compositions were evaluated: DDBSA 70.0 (a) Example 12, diluted with water to a 0.25% Cu-8- Propylene glycol methyl ether 26.0 Q content; Sugar cane seed was treated with this solution as (b) Example 18, diluted with mineral spirits to a 65 diluted with a water carrier. Seed also was treated with 0.25% Cu-8-Q content; benomyl, triophanate methyl, Panoctine (9-aza-1,17 (c) Cunilate 2174, diluted with mineral spirits to a diquanidinoheptadecane triacetate), hot water and cold 0.25% Cu-8-Q content; and water. The results are tabulated below: 4,766,113 25 26 which otherwise would remain in true solution in wa Concentration, ppm % Emergency ter. In some instances of use, it is desirable to maintain of active ingredients (1) Inoculated (2) the true solution character of the diluted composition. It DDBSA/2-(methoxy 50 48 has been found that a number of modifications to the carbamoyl)-benzimi 100 54 5 composition will either eliminate or delay the formation dazole 200 53 Thiophanate methyl 50 55 of solid matter in the use solution, the presence of such 150 51 particulate matter being evidenced generally by a 300 49 cloudy appearance. Among the more effective of such Panoctine 50 48 modifying agents are (1) partial or total replacement of 100 48 10 200 53 the diluting water with a lower alcohol such as metha Benonyl 150 50 nol, ethanol or isopropanol, or (2) additional of nonionic Hot water 0 36 surfactants such as alkyl phenol ethoxylates. Cold water O 27 The following compositions were prepared: (1) The active ingredient for the composition of this example is stated in terms of the benzimidazole content; all others are stated in terms of the listed chemical com 15 pound. (2) Prior to planting, the seed was inoculated with the causal disease organism, A. B Ceratocystis pilifera, Emergence data was collected 56 days after planting. Oxime 2.07 0.72 Copper hydrate 0.70 0.25 These results demonstrate the in vivo efficacy of the DDBSA 40.00 14.00 compositions of this example. Additionally noteworthy 20 Isopropanol 20.00 7.00 is the fact that, at the 27-day mark, the emergence rate Distilled water 37.23 5203 was noticeably higher for the composition of this exam Octyl phenol ethoxylate - 26.00 ple as compared to all other treatments. (9 EO) Example 52 25 The foregoing examples involve complexes between Composition A exhibited solution clouding when the disubstituted aryl compound and known antimicro diluted 1:300 with water containing 120 ppm hardness bials. Another feature of the invention is the discovery (total dissolved solids), whereas Composition B did not. that some agents not ordinarily considered to be antimi In terms of the Cu-8-O/DDBSA content, a 1:300 dilu crobial are effective when dissolved in the disubstituted 30 tion of Composition B is the equivalent of a 1:857 dilu aryl compounds of this invention. A case in point is tion of Composition A. phthalimide which is not offered commercially as a fungicide and which Horsfall, in Principles of Fungicidal Example 54 Action, describes as having very little fungicidal activ The alkyl aryl sulfonic acids of this invention, typi ity. Yet the composition described below exhibits excel- 35 fied by DDBSA, are strong acids and, as such, are lent control of stain and mold fungi in green lumber. harmful to many substrates and, in particular, are highly Phthalimide 0.44 lbs. corrosive to human skin and capable of producing se DDBSA 0.88 lbs. were irritation and burns. The addition of metal-oxine Sodium hydroxide 0.44 lbs. compounds to DDBSA, as described herein, reduces Water 100.00 gallons 40 the corrosive action of DDBSA significantly, even in The sodium hydroxide was dissolved in one quart of small amounts. This is entirely unexpected and no ade water, the phthalimide then added and the mixture quate explanation for this fact has yet been found. stirred until a clear solution resulted. The DDBSA was The following compositions were prepared (all parts then added with stirring, followed by the balance of the by weight): water. The composition was evaluated by treating 45 green, rough-sawn southern yellow pine boards. After two weeks' exposure: A. B C D Oxine 8.2 4.1 2. m Copper hydrate 2.8 1.4 0.7 m % Mold and Stain 50 DDBSA 59.0 59.0 40.0 59.0 DDBSA/phthalimide solution O Propylene glycol 30.0 35.5 57.2 40 Untreated control boards 45 methyl ether It is theorized, and indicated by Horsfall, that phthal These compositions were tested for skin irritation by imide is an active toxiphore, yet is essentially ineffective 55 rubbing 2 ml of each onto the hands of six test subjects. as an antimicrobial because of its low fat solubility. After 60 seconds, Compositions A, B and C were rinsed According to the concept of this invention, when a off the hands with water. None of the test subjects solution is formed between phthalimide and the disub noted any adverse effect such as stinging, burning, itch stituted aryl compound, the oleophilic substituent may ing or warming during the test or thereafter. The results lead the toxiphore into the microbial cell and act as an with Compositions D and E were observably different. active antimicrobial. Other active toxiphores, such as With 5 seconds after application, all test subjects noted benzimidazole, which lack only fat solubility for antimi a strong heating, burning sensation that required imme crobial activity, can be converted by this same concept diate and thorough removal with running water. A into active antimicrobials. relatively mild stinging sensation was noted thereafter Example 53 65 by four of the six test subjects for periods of about thirty minutes to two hours. High water hardness and/or high use dilutions can The compositions of this invention exhibit a unique cause precipitation of the compositions of this invention combination of cleaning effectiveness and mildness 4,766,113 27 28 heretofore unknown in cleaning agents. More specifi the antimicrobial activity below a usable level. Among cally: the types of usable and desirable other ingredients are Effectiveness-Equal to superior to the best tradi wetting agents, detergents and surfactants, emollients, tional types of cleaning agents. Cleaning efficacy, par other pesticides, thixotropes, film-formers, foam-build ticularly notable on metals, concrete, brick, porcelain ers and antifoamers, soil antideposition and antiredepo and grouting, is applicable both to inanimate substrates sition agents, wet slip enhancers, fillers, colorants, taste and human uses. modifiers, odor masking agents and fragrances, fire Mildness-Even at concentrations much higher than retardants, corrosion inhibitors, chelators for undesir normal use dilutions, no adverse effects upon the hose able metals including those causing water hardness, and substrate are noted when employed in typical cleaning 10 acidification and alkalization agents. Generally, the procedures. Such substrates include painted surfaces, compositions of this invention must be maintained, at most metals, concrete, brick, porcelain enamel, wood, the desired use dilution and at the time of use or incor plastics, wool, leather, glass, cotton, linen, synthetic poration into another composition, material or product, fabrics and the like. This unusual mildness and lack of at a pH of about 3.5 or below. The preferred pH, at least irritation is observed also in human uses, even upon the 15 when metal oxines are used, is below 2.8. most delicate skin areas, eyes and mucous membranes. The antimicrobial compounds of this invention are On dilicate body surface areas, the compositions proved effective to control odors caused by biological activity. significantly less irritating than Ivory Soap, a long-time Since many of the compounds become affixed to many standard for mildness. substrates, their value as an antimicrobial and order High cleaning efficacy for general purpose uses cou 20 destroyer will remain for a prolonged period of time pled with unusually low substrate damage potential to and even survive many washings of certain substrates, inanimate substrates and lack of irritation to humans and such as garments. animals is uncommon in low pH products and unusual The physical form of the compositions of this inven even in conventional cleaners in the relatively neutral tion can vary widely. The most commonly desirable pH range (6 to 8). By comparison, the pH of use concen 25 form is a low viscosity liquid-the form of the examples trations of the composition of the invention most gener given above. But other forms fall within the scope of ally range from about 1.5 to about 2.5. the invention and may be preferred for certain end uses, Historically, cleaning agents have been alkaline, and including solids, powders, high viscosity liquids, oint the higher the alkalinity, the greater is the effectiveness ments and creams, and gels. All of the foregoing exam and harshness. In recent years, a trend toward mildly 30 ples consist of compositions that are true solutions, both acid chances (pH 5 to 6) for certain specialty uses-hair in concentrate and use-diluted form. Such true solution shampoos, for example-has been underway, and compositions generally are preferred for the majority of highly acidic and oxidizing cleaning materials are tradi cases for a number of practical handling and application tional for certain highly specialized applications such as reasons. However, compositions that are in emulsion metal, dairy equipment and toilet bowl cleaning. In 35 form or which exist as liquid or solid suspensions are these cases, the acidity serves a specific purpose, such as within the scope of this invention and have definite removal of oxide (metals), hardened milk deposits (dair utility. ies) and iron discoloration (toilet bowls). Such products, The compositions described above will hereinafter be however, damage most substrates and are much too referred to as the "compositions of section l'. corrosive for human use. The low pH compositions disclosed herein not only are contrary to traditional SECTION 2 concepts, but exhibit an extremely broad range of con We have also discovered that the antimicrobial activ sumer, commercial and industrial uses ranging from a ity of known phenolic antimicrobials can be improved vaginal douche at one extreme to cast iron engine block by mixing a minor amount of such known phenolic cleaning at the other end of the spectrum. In certain antimicrobials with a major amount of a solution pre specialized applications where highly acid cleaners pared by solubilizing a non-phenolic antimicrobial have long been used, the compositions will provide agent as set forth in section I with a disubstituted aryl equal to superior cleaning action without the associated compound having a hydrophilic and an oleophilic sub dangers. For example, the compositions will clean most stituent (as described in section I hereinabove). In one leather surfaces more effectively than an oxalic acid/ 50 preferred embodiment said second antimicrobial agent water solution, a recommended leather cleaner which is is an organometallic compound (as described in section toxic, and corrosive to humans, animals and most sub I above) such as a metal chelate of oxine and the disub Strates. stituted aryl compound is alkyl benzene sulfonic acid It is within the scope of this invention to modify the (most preferably DDBSA in which the alkyl chain disclosed compositions to improve performance or alter 55 predominates in C10 to C13.) physical properties and meet final product needs for a Certain phenolic compositions are known to the prior wide range of end uses in many fields, including product art that have efficacy against certain microbials includ material and process preservation, medical, animal ing fungi and Gram-positive and Gram-negative bac health care, water treatment, plant disease control, dis teria. Many of these phenolics are falling into disfavor infection, sanitization and cleaning, cosmetics and toi because of their relatively high toxicity toward animal letries. and plant life. This is particularly true since, for many In many instances, the compositions of the invention applications, an undesirably large amount of phenolics constitute only a minor portion of the final product must be used to achieve a desired degree of antimicro formulation, in substantial measure because the high bial activity against certain microbes. The utilization of efficacy requires only small amounts. The rule for use of 65 phenolic antimicrobials is further limited in that they other ingredients in the disclosed compositions is simple are frequently limited in the spectrum of microbes and obvious-namely, such ingredients must be com against which they display effective antimicrobial activ patible with the compositions and must not deactivate ity. 4,766,113 29 30 Accordingly, a further object of this invention is to accordance with the aforementioned AOAC Use Dilu improve the efficacy of phenolic antimicrobial agents. tion procedure, with these results: A related object of this invention is to formulate phenolic antimicrobial compositions in a manner in which effective antimicrobial activity can be achieved Use Dilution No. of Tubes Tested with a minimum quantity of the phenolics, in Water Positive Negative" Another object of this invention is to widen the spec A. 1:500 9 trum of microbes over which phenolic compositions C 1:750 . O 10 display effective antimicrobial activity. (positive -growth; negative - no growth) These and other objects of this invention are 10 achieved by including a minor amount of an antimicro Reference tests on antibacterials and the manufactur bial phenolic composition in a solution comprised of a er's technical literature on 4-chloro-2-phenyl phenol non-phenolic antimicrobial composition as set forth in indicate that this phenol has very low activity against P. section I and a disubstituted aryl compound as set forth aeruginosa (a phenol coefficient of "less than 10'). Even in section I. 15 assuming a phenol coefficient of as high as 10, the This embodiment of our invention is illustrated by the amount of 4-chloro-2-phenyl phenol in Composition C following examples: should have improved, by calulation, the use dilution efficacy of Composition C by about 4% over that of Example 1 Composition A which contains no phenol. Instead an The following two compositions were prepared 20 improvement of an indeterminate amount in excess of 50% was obtained, indicating that the addition of the (amounts are weight percent): phenol to Composition A improved its antibacterial activity out of all proportion to the amount of phenol A. B added. 25 Oxine 2.05 2.05 The antifungal efficacy of these phenolics is im Copper hydrate 0.07 0.70 proved by formulation in accordance with this inven DOBSA 40.00 40.00 tion. Orthophenyl phenol, 2,3,5-trichlorophenol, tetra Isopropanol 32.00 32.00 Distilled water 25.25 15.25 chlorophenol and pentachlorophenol, for example, ex Orthophenyl phenol -- 10.00 hibit antifungal efficacy improvement significantly 30 higher than would be expected when formulated con ventionally, especially for use in a water carrier where Both compositions were tested against the Gram-pos customary formulation methods employ a water-soluble itive Staphylococcus aureus bacterium in accordance alkaline salt of such phenols, such as the sodium or with the AOAC Use Dilution Confirmation Method potassium salts. with these results: 35 The quantity of phenolic antimicrobial added to the antimicrobial solution is not critical and can be varied Use Dilution No. of Tubes Tested depending on the microorganism that it is desired to in Water Positive" Negative' control and the permitted toxicity of the total composi A 1:1000 O 10 tion. As noted in the examples, a 10% inclusion of a A. 1:1250 1 9 phenolic is quite effective and can vary over a wide B 1:1250 O 10 range, although 5-20% is generally preferred. B 1:500 1 9 ("positive -growth; negative - no growth) SECTION 3 We have also discovered a method for reducing the These results indicate that efficacy of Composition A 45 skin and eye irritation of alkyl arylsulfonic acids (as set was improved by about 20-25% by addition of 10% forth in Section 1). In a preferred embodiment, the orthophenyl phenol to Composition A (forming Com irritation-reducing agent is an organometallic antimi position B). By calculation, using technical bulletins crobial agent (as set forth in section I) which is solubi published by the producer of orthophenyl phenol for 50 lized in the alkyl aryl sulfonic acids and which in addi disinfectant formulation, the efficacy improvement tion provides for antimicrobial efficacy. The irritation should have been about 4%. In short, the orthophenyl of the solution can further be reduced by including a phenol addition actually improved efficacy by a factor second irritation-reducing agent such as polyvinyl pyr of five to six times the calculated theoretical improve rolidone or selected surfactants such as ethoxylated and Inent. 55 prothoxylated long-chain aliphatic alcohol type surfac tants, ethoxylated alkyl phenol type surfactants or a Example 2 sulfosuccinate of a higher fatty acid monoethanolamid. Composition C was prepared as follows (amounts are The following Examples are illustrative: weight percent): Oxine 2.05 Example 1 Copper hydrate 0.70 A commercial grade of undiluted DDBSA was DDBSA 40.00 poured onto and spread over the hands of several per Isopropanol 32.00 sons. An almost immediate warm and uncomfortable Distilled water 15.25 sensation was noted, causing the persons to flood their 4-chloro-2-phenyl phenol 10.00 65 hands with water to remove the DDBSA. Composition C was tested and compared with Com The experiment was repeated, using DDBSA into position A of Example 1 against the standard Gram which had been dissolved 10% by weight of copper-8- negative bacterium Pseudomonas aeruginosa PRD-10 in quinolinolate (Cu-8-Q). No immediate sensation of dis 4,766,113 31 32 comfort was felt as had been the case with the unmodi ples, using the same test procedures. These nonionic fied DDBSA. surfactant types do not appear, however, to be as effec tive as PVP, but have proved particularly useful for Example 2 many applications where the detergency characteristics The following composition was prepared: 5 Oxine 2.05 parts by weight of these additives to the basic compositions of the inven Copper hydrate 0.70 parts by weight tion are desirable. Examples of useful surfactants in DDBSA 40.00 parts by weight these classes include phenols with C8 through C12 alkyl Isopropanol 15.00 parts by weight side chains, alcohols with C10 to C22 chain lengths and Water 42.75 parts by weight 10 ethylene oxide and propylene oxide adducts thereon A second test solution also was prepared wherein five from about 4 to about 100. Suitable alkyl phenols in parts of the water in the above composition was re clude octyl phenol, nonyl phenol and dodecyl phenol placed by an equal amount of polyvinyl pyrrollidone plus dinonyl and didodecyl phenols. (PVP) solution in water (GAF's NPK-60). By way of example, the first composition of Example The two compositions were tested for skin irritation 15 2 was prepared except that 20 parts of water were re by placing two drops of one composition on the skin at placed with an equal amount of octyl phenol ethoxylate the inside of one elbow of five test subjects and two (9 EO). Tested as before, this composition exhibited drops of the other composition on the same spot on the distinctly less skin irritation than without the octyl phe other elbow of each subject. In all cases, the composi nol ethoxylate. tion was smeared over about a one-half square inch of 20 Another additive found to reduce the irritation of the skin and the subjects' reactions were noted over a 20 compositions of the invention is a sulfosuccinate of a minute test period. higher fatty acid monoethanolamid sold by Cyclo The first composition (no PVP) was described vari Chemicals Corporation under the designation Cyclopol ously as "burning,' 'stinging,' "tingling' and "warm SBR-3. Significant improvement in irritation character ing". The second composition (containing PVP) was 25 istics was observed using 3%, 10% and 20% replace described by the subjects over the range of noticeably ments of water in the first composition of Example 2 milder skin sensation to none at all. with this material, which has a 40% active ingredient A third composition was prepared, identical to the content. second except that the PVP content was increased to It should be noted that no critical lower or upper twenty parts, again replacing an equal amount of water. 30 limits have been noted as to the amounts of these irrita Tested as before, no skin sensation was reported by any tion reduction agents which may be used in the practice of the test subjects. of this invention. Generally, increased amounts pro Example 3 duced greater reductions in irritation. The quantity added may be varied not only to obtain the desired These compositions were prepared (all parts by 35 irritancy reduction, but also to obtain other desired weight): properties in the final formulation. SECTION 4 A. B Oxime 4. 4.1 We have also discovered a method for materially Copper hydrate 1.4 1.4 40 increasing the solubility of compounds having insecti DDBSA 60.0 60.0 cidal properties. Other compounds heretofore recog Isopropanol 34.5 24.5 nized as having only antimicrobial activity become PVP (NPK-60) - 10.0 useful for insecticidal purposes when solubilized, partic ularly through the use of a disubstituted aryl compound The compositions were tested on three subjects for 45 described in Section 1, particularly an alkylbenzene or skin irritation by spreading two drops of each composi naphthylene sulfonic acid. tion over a one square inch area of skin just below one Thus, our invention also includes insecticides which eye of each test subject and the other composition on may be made water-soluble, at least in concentrated the same spot beneath the other eye. form, and which may, in some of the formulations dis Composition A became uncomfortable on the three 50 play reduced toxicity towards mammalian and plant test subjects within one to three minutes and was re life. moved. Composition B exhibited no sensation of dis The compositions set forth in section I have been comfort to much milder sensation than Composition A found to exhibit an unexpected degree of insect control. for the ten-minute duration of the test. This has importance in a variety of end uses, notably wood preservation, wherein there is little advantage in Example 4 55 protecting wood from fungal decay if the wood is pre The first composition of Example 1 was prepared served only for insects, such as termites, to destroy. In containing 30% PVP solution. When tested in accor many areas of the world, including those with high dance with the procedure of Examples 1 and 2, no sen decay rates, destruction of wood by termites occurs sation of skin discomfort was noted by any of the test 60 more rapidly than does fungal degradation. subjects. It also has been found that the compositions set forth in Section 1 are compatible with a broad range of well Example 5 known insecticidal chemicals in use for abroad range of It further has been discovered that a range of ethoxyl agricultural, medical, animal health care, product and ated and propoxylated long-chain aliphatic alcohol type 65 material preservation, and micellaneous pest control surfactants and ethoxylated alkyl phenol type surfac applications. This discovery extends the end use range tants provide noticeable skin irritation reduction for of the compositions of Section 1 where insect control in compositions of the type disclosed in previous exam addition to that afforded by the composition itself is 4,766,113 33 34 desired, or in those instances where the basic composi An important area of use for the insecticide-antimi tion provides no control of the target pests. Providing crobial compositions described in this section relates to insect and antimicrobial control in one composition also has both economic and convenience advantages. the known usefulness of certain insecticides for protec Entirely unexpected is our discovery that a number tion of wood in marine service against marine borers of standard insecticides incorporated in the composi which attack and destroy wood rapidly. There are three tions of Section 1, including DDBSA alone or in combi important species of wood-destroying borers in two nation with an antimicrobial, form true solutions in the classes: (1) Molluscan-Teredo and Martosia sp., and formulation concentrates and, of far greater signifi (2) Crustacean-Limnaria sp. It is near-axiomatic that cance, true solutions when water-diluted for use. Most O no one toxicant is effective against all three species, and insecticides are water-insoluble and consequently must it is a fact that in certain tropical waters, no single wood be formulated for use as wettable powders, flowables, impregnant has provided satisfactory durability. For or as emulsifiable liquid concentrates. Generally, these example, some waters off Key West Florida are inhab products, when diluted for use with water as the carrier, ited by combinations of marine borers which will de form suspensions or emulsions of mediocre stability, 15 stroy a wood piling impregnated with 20 lbs of creosote requiring frequent agitation to maintain an acceptable per cubic foot in five years, despite the fact that heavy degree of dispersion in the carrier. True solutions are creosote loadings in wood are considered to be the easier to use and provide more uniformity of coverage preservative of choice for marine applications the of the substrate or host organism for protection against world over. Additions to creosote of chlordane, diel the target organisms. True solutions also make practical 20 drin, and TBTO among other insecticides is reported to certain end uses where suspensions and emulsions of inadequate stability and poor substantivity are not func substantially improve creosoted wood piling durability tionable. Such suspensions and emulsions, for example, in certain waters. will not penetrate wood adequately. For long-term Accordingly, addition of these insecticides to the protection of wood for many uses, deep impregnation is 25 basic antimicrobial composition of Section 1 provides mandatory. improved wood durability in marine environments. A number of additional standard insecticides form Additionally, our discovery allows use of suitable insec true solutions when incorporated into the compositions ticides in a water carrier for impregnation of wood for of Section 1 in concentrated form, but form-stable col the first time. loidal emulsions when water-diluted for use. Such emul 30 Many of the known insecticide compounds presently sions for most uses offer all of the advantages of true available are objectionable in varying degrees due to water solutions in terms of desired stability and ability the toxicity they display toward animal and plant life. to penetrate and adhere to substrates such as wood and This is in distinction to the antimicrobial compounds of tight-weave fabrics. Section 1, which at use dilutions, are generally consid While water generally is the carrier of choice for 35 ered non-toxic. Thus, when both toxicity and efficacy preparing dilutions, other carriers have utility in certain are important, it is possible to use the antimicrobials in instances, including hydrocarbons and a variety of polar combination with known insecticides in order to sub organic solvents. stantially retain effectiveness while reducing the A criterion for combining insecticides with composi amount of the toxic insecticides that must be used. tions of the basic invention is stability of the insecticide to acidic media. The amount of insecticide practically The examples which follow demonstrate the uses and incorporatable into composition concentrates for subse practice of our discoveries: quent use dilution in true solution or stable emulsion Example 1 form depends upon a number of factors including inher ent solubility characteristics of the insecticide in the 45 The composition below was prepared and tested for basic composition of Section 1 and the make-up of this protection of wood against termite attack (amounts are basic composition. The larger the amount of DDBSA in parts by weight): said basic composition the larger the quantity of a given Oxine 4.1 insecticide that may be solubilized in the basic composi Copper hydrate 1.4 tion. The ratio of insecticide to said basic antimicrobial 50 DDBSA 65.0 composition is relatively noncritical and the choice in Methanol 29.5 large measure depends upon the desired combination of The test procedure was AWPA (American Wood antimicrobial-to-insecticidal performance. Preservers' Association) Methods M12-72, revised Among the useful insecticides that may be incorpo 1973. The termite species employed was Reticulitermes rated into the solubilizing disubstituted aryl compounds 55 flavipes. The test blocks were southern pine sapwood of Section 1, either with or without antimicrobials ac and the test period was four weeks. cording to Section 1, are carbaryl, methoxychlor, lin dane, chlordane, malathion, parathion and methyl para thion, toxaphene, dieldren, aldrin, endrin, heptachlor, Composition Retention in Wood (pcf)" % Weight baygon and cygon. TBTO (tributyl tin oxide) which is Total Composition Cu-8-0 Loss not a recognized insecticide, also has utility against 0. O 10.1 marine wood borers. 0.38 0.20 0.1 In cases where antimicrobial action is not wanted, it is 0.62 0.033 0.6 within the scope of this invention to incorporate the 0.84 0.045 O insecticide in the disubstituted aryl compounds of Sec 65 tion 1 plus a diluent. However, some insecticides seem These data indicate that the test composition provides ingly exhibit reduced solubility characteristics in the adequate protection against termites at the lowest con absence of a metal-oxine chelate according to Section 1. centration evaluated. 4,766 ,113 35 36 The composition as prepared formed a true solution. Example 2 When diluted 1:99 with water, a true solution resulted. The following composition was prepared (amounts are parts by weight). The composition was a true solu Example 7 tion. When diluted 1:9 with water for use, a true, stable 5 The following compositions were prepared (amounts solution resulted. are parts by weight): Oxine 7.4 Copper hydrate 2.5 DDBSA 53.0 A B Chlordane 10.0 O Oxine 4. 4, Copper hydrate 1.4 1.4 Propylene glycol methyl ether 27.1 DDBSA 65.0 65.0 Malathion 5.0 m Example 3 Methyl parathion 5.0 The following compositions were prepared (amounts Methanol 24.5 24.5 are parts by weight): 15 Both compositions produced true solutions as pre A. B pared and when diluted 1:100 with water. Oxine 7.4 7.8 Example 8 Copper hydrate 2.5 2.7 20 DDBSA 53.0 53.0 The following compositions, all dilutable for use with TBTO 10.0 5.0 either water or petroleum hydrocarbon solvents, and Propylene glycol methyl ether 27.1 31.5 diluted in the range of 1:9 to 1:19, provide excellent marine and land service protection to wood when im Both formed true solutions are prepared. Diluted 1:9 25 pregnated into the wood at a 20-25 pcfretention. with water, Composition. A formed a stable colloidal emulsion. Diluted 1:9 and 1:99 with water, Composition B formed true solutions. l 2 3 4. 5 6 7 8 Oxine 4, 8 4. 8.2 4.1 40 4.0 8.2 4 Copper 3 7 1.4 2.8 - - m 2.8 l Example 4 30 hydrate The following compositions were prepared (amounts DDBSA 60 60 600 60.0 60.0 60.0 60.0 60.0 60 are parts by weight): Stannous -- - m- -- 7.0 4.5 4.5 - Sulfate Chlordane 10 - m - - 10.0 - - Toxaphene - 5 5.0 - - - 6.0 - A. B 35 TBTO --- - - a - 10.0 10 Oxine 7.4 7.8 Methanol 23 20 29.5 29.0 28.9 21.5 25.5 19.0 24 Copper hydrate 2.5 2.7 DDBSA 53.0 53.0 Toxaphene 10.0 5.0 Propylene glycol methyl ether 27.1 31.5 Example 9 40 The following antifouling compositions (to reduce Both produced true solutions as prepared. Composi barnacle and other marine organism encrustation of ship tion. A formed a stable colloidal emulsion when diluted hulls), suitable as additives to marine hull coatings, were 1:9 with water. Composition B produced 1:9 and 1:99 prepared. The amounts listed are parts by weight: dilutions in water with a slightly hazy appearance, indi 45 cating a borderline condition between a true solution l 2 3 and a colloidal emulsion. Oxine 9.0 9.0 15.0 Example 5 Copper hydrate 7.0 - 10.0 Stannous sulfate 7.5 - The following composition was prepared (amounts DDBSA 60.0 60.0 40.0 are parts by weight): 50 Methanol 24.0 23.5 35.0 Oxine 4.1 Copper hydrate 1.4 These compositions can be incorporated in a number DDBSA 65.0 of marine coating formulations in amounts up to 15%. Malathion 5.0 55 Conventional antifouling components, such as cuprous Methanol 24.5 oxide, may be used to augment the protection against As prepared, the composition was a true solution. fouling organisms. Dilution 1:99 with water for use also produced a true solution. Example 10 The following compositions form true solutions in Example 6 water as prepared, and true solutions when diluted The following composition was made (amounts are 1:100 with water. parts by weight): Oxine 4.1 A. B C D Copper hydrate 1.4 65 DDBSA 65.0 DDBSA 60 60 60 60 Chlordane 5 - -- - Carbaryl 5.0 Toxaphene m 5 ------Methanol 24.5 Melathion m - 5 - 4,766,113 37 38 Asphalt roofing-Both the asphalt and the cellulosic -continued felt core in shingle and roll roofing are degraded by a A. B C D variety of fungal species. The first major indication is Methoxyclor --- u- 5 product embrittlement which also is the standard crite Methanol 35 35 35 35 rion of roofing failure. The compositions of Section 1 have unique utility by reason of high substantivity, In general terms, the diphenyl, carbamate and chlori resistance to water extraction, and heat resistance. nated cyclo organic compound classes of insecticides Roofing is a severe long-term test of these properties. function well in this invention. A number of insect con Plastics-A substantial number of formulated poly trol compounds in the organic phosphate class perform O mers for a wide range of interior and exterior applica satisfactorily although stability time usually is reduced tions are subject to fungal deterioration and aesthetic compared to the preceding classes. disfigurement, from bathroom shower curtains to exte rior building panels. The compositions of Section 1 SECTION 5 offer unusual heat stability for these uses. We have also discovered that the compositions of Textiles and fiber products-These products, includ Section 1 (comprising said antimicrobial agent and said ing rope, netting, yarns, twine and carpeting, and partic disubsituted aryl compound) are useful for protecting ularly those made from natural products such as cotton, various manufactured goods and industrial commodities hemp, linen and wool, generally are more suceptible to from microbial attack or degradation. The compositions biological breakdown than the synthetics although the disclosed in Section 1 exhibit broad spectrum antimicro 20 latter are not immune generally. Fungal deterioration is bial efficacy against microorganisms which degrade the most common, except in the case of wool which is materials of economic value. The range of activity and highly subject to bacterial degradation. Protection from high per unit efficiency covers all classes of fungi and microorganism attack is difficult in severe service con both Gram-positive and Gram-negative bacteria. Cou ditions because of the very rapid rate of product failure. 25 The compositions of Section 1 are uniquely suited for pled with a number of auxiliary properties which have many such preservative uses because of high unit effi few undesirable side effects during application or in cacy, low toxicity, low skin irritation, and resistance to service, said compositions have unexpected and unique actinic ray deactivation of antimicrobial efficacy. versatility for preservation of a wide range of materials Wood disinfection-Wood is employed in a number and products. Heretofore available high-efficacy, 30 of applications involving direct (butcher blocks) and broad-spectrum antimicrobial agents have one or more indirect (pallets) contact with a broad range of food of a wide range of undesirable properties that can se stuffs. The porosity of wood and other factors makes verely limit their utility. Among the frequently encoun disinfection, often a mandatory requirement, very diffi tered limitations are mammalian toxicity, odor and cult. Complicating this problem is the need for safety color, skin and eye irritation, volatility and instability, 35 and low odor, taste and color in the antimicrobial con low persistence, and narrow antimicrobial spectrum. position. The compositions of Section 1 uniquely fit The compositions of Section 1 eliminate these and other these needs and, in addition, provides uniformity of bars to usage. penetration into the substrate. The combination of properties of said compositions Water flood oil recover-Bacteria introduced with meet the specialized needs of a broad and diverse range water for secondary oil field recovery may proliferate, of material preservation applications, including both clogging porous oil sands and initiating corrosion of finished products and in-process manufacturing steps. pipe and other equipment. The two most important Among the suitable uses providing one or more unique organisms, Desulfovibrio desulfuricans and Thiobacillus results, the following may be given as representative: suboxydans, are controlled with high efficacy by the Leather-The compositions of Section 1 have high 45 compositions of Section 1 which also provide the eco substantivity to protein substrates, low toxicity, low logical safety that is desirable in this use because of the irritation to human and animal tissues, and high antimi enormous quantities of water involved. crobial efficacy against the fungal, and especially the Petroleum fuels-A number of fungal genera, includ bacterial, organisms that deteriorate leather. ing Hormodendrum, and bacterial genera, including Paints and adhesives-Water-based paints and adhe 50 Pseudomonas, Bacillus and Aerobacter, grow abun sives require an antibacterial and antifungal agent for dantly at the water/oil interface of many petroleum in-can preservation. Oil-based paints for exterior service fuels causing slime deformation and metallic corrosion. frequently include antifungal agents. The compositions This may have especially grave consequences in critical of Section 1 possess an unusual degree of appropriate fuel uses such as for jet aircraft engines. The high effi antifungal and antibacterial action relative to persis 55 cacy of the compositions of Section 1 plug long-term tence of action after application. Heretofore antibacte antimicrobial stability is of substantial value for fuel rial action has been limited to in-can prevention of prod preservation use. uct deterioration and to exterior service control of fungi Chemical toilets-The high efficacy of the composi that aethetically disfigure paint and cause its deteriora tions of Section 1 plus low environmental contamina tion. tion are of special value for such devices as planes, Cutting oils and metal drawing compound-These trains, boats, buses and compers, as well as in stationary water-based materials require antifungal and antibacte uses, for chemical toilets. rial protection to prevent rancidity, slime development, Logs-Logs for lumber and paper conversion fre and possible growth of organisms pathogenic to man. quently are stored under water (water-sprayed piles or The compositions of Section 1 provide the needed 65 in ponds) for protection against fungal degradation. broad spectrum antimicrobial activity with an unusual While generally useful, this procedure has limited value degree of safety to metal fabrication operating person because high moisture content in the wood does not nel. inhibit growth of many bacterial and certain soft-rot 4,766,113 39 40 fungi, and because of the considerable difficulty, espe are relatively nontoxic to mammals and have low irrita cially with water-sprayed piles of logs, in achieving tion to mammal tissue. The compositions of Section 1 all-over water/log contact. The compositions of Sec are relatively low in color, odor and taste; have long tion 1 are uniquely useful for addition to the water stability under normal ambient conditions; have low because they have the necessary high broad-spectrum volatility; are heat and light stable; can be diluted for efficacy and they are environmentally safe. use in many carriers; become insoluable and substantive Breweries-The brewing of beer is a microbiological to a host upon application; and are effective cleaning process with high susceptibility to ruinous results from and wetting agents. even minor contamination with stray organisms of While it is believed necessary in the practice of this many types. The broad-spectrum, safe effectiveness of 10 invention to form an initial solution of the antimicrobial the compositions of Section 1 provide unique equip agent with the disubstituted aryl compound (as these ment protection, and the persistence of antimicrobial are defined in Section 1) the antimicrobial agent may, activity is of great value in the dual area of fungal and when diluted for use, begin to form a very fine precipi bacterial control on walls, ceilings and floors in the tate. This precipitation from solution may and usually vicinity of storage, processing and handling facilities. 15 will become complete after the composition has been applied to a substrate. SECTION 6 The compositions of Section 1 eliminate overgrowth We have also discovered that the compositions of by reason of broad-spectrum control of all major types Section 1 are useful for the treatment of mammalian of microbiological flora. This range of activity is both diseases. microbiostatic and microbiocidal, in contrast to many The compositions of Section 1 are generally distin present products which are effective primarily as bio guished from the useful antimicrobials known to the stats. The activity is both initial and residual, exhibiting prior art in that they exhibit a high efficacy against a both rapid initial organism kill or inhibition, depending broad and non-selective range of numerous types of upon the concentration of active ingredients employed, microorganisms that are pathogenic to man and ani 25 and considerable residual persistence of antimicrobial mals. These microorganisms include both Gram-posi effectiveness. tive and Gram-negative bacteria, the four primary The compositions of Section 1 combine a number of classes of fungi (Ascomycets, Basidiomycetes, Phyco auxiliary properties that not only are prerequisites for mycetes and Fungi Imperfecti), yeasts envelope (lipoid) human and animal use, but which, in total, are unique in and naked (hydrophilic) viruses, protozoa and helmin 30 the field of antimicrobials. These include: thes including mematoda and trematoda. Broad spec Low mammalian toxicity. The compositions of Sec trum antimicrobials that are safe for human and animal tion 1 have much lower toxicity than the majority of use and have useful efficacy against Gram-positive and anticicrobials in use today and have been given labora Gram-negative bacteria and fungi are relatively rare tory ratings of "not toxic orally.” and essentially unknown in the field of antimicrobials, 35 Low skin and eye irritation. The range of skin and Broad spectrum antimicrobials are highly significant eye irritation is from low for concentrates to low to in overgrowth control. Since various types of microor zero in dilute concentrations effective for use. Opthal ganisms compete with each other for survival, both mic preparations containing the compositions of Sec inside and outside the mammilian body, many widely tion 1 for antimicrobial eye medication exhibit no eye employed narrow-spectrum antimicrobials which con irritation. trol only selected organisms, provide an environment Low color, odor and taste. Concentrates of the com for the growth of other organisms. This is of notable positions of Section 1 exhibit a low inherent odor level, concern to the medical profession in the widespread and impart minor to no color to most substrates. At consumer use of medicated soaps and other prepara suitable use dilutions, such compositions impart no tions for the topical application in which, as a rule, the 45 color, no odor, and very low to zero taste. It therefore antimicrobial agents used in the preparations have prac is generally practical in formulated products containing tical effectiveness essentially only against Gram-posi the compositions of Section 1 to incorporate colorants, tive bacteria. This can and does allow establishment and fragrances or masking agents, and taste flavoring agents proliferation of Gram-negative bacterial and fungal as may be desired. pathogens not usually present on the skin, which organ 50 Shelf stability. No significant changes take place in isms can be considerably more dangerous to the host antimicrobial properties in the compositions of Section than Gram-positive bacterial that are brought under 1 even after long storage periods under normal ambient control. By way of specific example, many of the com conditions. mon preparations will, with continued use, encourage Low volatility. Use dilutions of the compositions of the growth of the Gram-negative Pseudomonas sp., 55 Section 1 maintain initial efficacy upon standing in open which is difficult to control even with antibiotics. containers for long time periods. Evaporation of carri Many known antimicrobials for use in treating mam ers, usually water, will increase efficacy by raising the milian disease are also deficient in that they have rela ratio of composition to carrier in the use dilution. A tively high toxicity and may have such high irritability number of presently used antimicrobials require special to animal tissue as to render them practically of little use precautions because of volatility or sublimation use. These problems, particularly that of toxicity, are processes. assuming particular importance in more recent times as High air, heat and light stability. In contrast to a various health and governmental agencies place in number of present products, the compositions of Sec creasingly stringent requirements on antimicrobials tion 1 exhibit no deterioration or loss of efficacy from before permitting their use. 65 prolonged exposure to air or light. Heat stability up to It has been found that the compositions of Section 1 an estimated 300 F. allows a range of use conditions are efficacious against a broad and non-selective range not feasible with many currently available antimicrobi of microorganisms that cause mammilary disease, and als. Heat plus inherent antimicrobial action of the com 4,766,113 41 42 positions provide a flexibility of use and performance crobials. Such overgrowth can be both rapid and dan unmatched by any narrow-spectrum antimicrobials. gerous. This increased flexibility is particularly useful in that no Surgical antiseptics. The compositions of Section 1 special equipment is needed for utilization to offset provide extremely high skin degerming capacity as volatility, light or air instability, active ingredient subli measured by reduction in the transient and resident mation or the like. flora count as shown in Example. It is believed that, in Carrier versatility. While water is the most widely addition to the demonstrated high antimicrobial effi used and versatile use diluent for antimicrobials, the cacy, the compositions of Section 1 may have excep compositions of Section 1 may be carried in a very tional capacity to act upon the difficult to reach flora broad range of polar (including water) and non-polar 10 under the outer, cutaneous layer of skin which consists solvents, and combinations thereof. Carriers other than of the flat plaques of keratin. In view of the generally water may be very useful in a variety of end uses for a held belief that total degerming is not possible short of number of reasons including enhanced antimicrobial destruction of the skin itself, the greater the reduction of action, need for rapid carrier evaporation, general un flora, the lower the risk of the patient from a surgeon suitability of water, need for low or non-volatility of the 15 who is compelled to operate with hands that cannot be carrier, the desire for carrier lubricity or emolliency, fully disinfected and to make incisions through skin that the adverse effect of water upon the host organism or cannot be made entirely germ-free. The exceptional substrate and the like. Suitable carriers include a broad flora count reduction afforded on the skin by the com range of hydrocarbons, alcohols, ketones, ethers, gly positions of Section 1 is unexpected in view of the long cols, chlorinated organic solvents, esters, acetyls, phta 20 known difficulty of the skin degerming and the ac lates, adipates, aldehydes, anhydrides, acids, silicone knowledged fact that, as the count reduction ap fluids, nitroparaffins plus a variety of the more exotic proaches the theoretical 100% maximum, the problems solvents such as N-vinyl-2-pyrrolindone, dimethylsulf of achieving even a very small incremental improve oxide and dimethyl formamide. This carrier versatility ment in the count becomes progressively more difficult is in sharp contrast with most antimicrobials which are 25 by orders of magnitude. either imcompatible with, or are insoluble in, many Another reason for desiring to achieve the highest types of carriers. reduction in skin count that is feasible, both on the Insolubility after application. For general use, the surgical team and the patient, is the fact that surgical preferred embodiment of compositions of Section 1 are gowns, gloves, patient draping, folds in the skin, the soluble in the use-dilution carriers. Such ready-for-use 30 perineal and umbilicus areas and the like provide both compositions, notably in a water carrier, become water protectin and favorable (warm and moist) conditions insoluble after application to the host. This is in sharp for rapid and accelerated growth of the remaining or contrast to most antimicrobials which retain water solu ganisms. Hence the high order of initial flora reduction, bility after application. This feature offers obvious re and the substantivity and persistence of antimicrobial sidual persistence advantages since removal by aqueous 35 activity of the compositions, has important utility when media is a major means by which initial effectiveness of consideration is given to the fact that containment of antimicrobial agents is rapidly diminished. the rapid regrowth generally is very difficult. For in Substantivity. The compositions of Section 1 not only stance, perforation of gloves worn by the surgical team turn water insoluble rapidly after application as just 40 is a frequent occurrence with numbers as high as 70% mentioned, but also exhibit strikingly high affinity for a being reported. With glove punctures or tears, flora number of surfaces, most notably cellulosic and protein ladened sweat and the like may leak large numbers of aceous substrates. This is true under usual parameters of microorganisms into a surgical wound. use. It normally occurs near-instantly and in the wet The ultimate in degerming effectiveness of the com condition. The result for a number of uses is unusually 45 position is obtained by means of an after-application of high retention of antimicrobial action yielding surpris the composition following the surgical scrub or patient ingly long efficacy life. preoperative skin preparation. This may be a rinse in a Wetting-cleaning action. The compositions of Sec water solution of the composition, or an alcoholic tinc tion 1 possess strong, inherent wetting and detergent ture, or most effective of all, an ointment carefully activity. This feature is a significant aid in penetrating 50 rubbed into the skin, under the fingernails and into crevices and areas of difficult access. Further, the high protective folds of the skin and mucous membranes. For detergent-cleaning action is an important adjunct for the hands of operating personnel, an ointment which many medical antisepsis and disinfection applications. leaves the hands non-slippery can be readily formu Efficaceous in the presence of organic matter. Unlike lated. Not only does such an ointment or cream present many antimicrobial compositions such as a number of 55 physical barrier of sorts to the movement of microflora, the quaternary amines, the efficacy of the compositions but much more importantly, a finite layer of the highly of Section 1 are not destroyed by the presence of or toxic composition is interposed between the organisms ganic material. remaining after the degerming step and the surgical The following is an incomplete, but representative wound or any other area of the patient and surgical listing of uses for the compositions of Section 1 which 60 personnel subject to pathogenic contamination. uses are unique in kind or degree of effectiveness. A Where desired, the compounds clean as well as disin feature common to these uses is effective control of an fect. The cleaning action can be applied to unbroken uncommonly large number of the major classes of paho skin, wounds and body cavaties. gens as previously described. This feature has primary Disinfection of medical and surgical materials. The importance where kill or inactivation is mandatory and 65 compositions of Section 1 are suitable for disinfection of in uses where non-selective control of all organisms is critical items, such as scalpel blades, transfer forceps desirable to prevent overgrowth of pathogenic flora and cardiac catheters; semi-critical items such as ther otherwise not controlled by selective-spectrum antimi mometers, aspirator tubes and cystoscopes; and non 4,766,113 43 44 critical items such as face masks, rebreathing bags and a which antibotic treatment is relatively ineffective. Con variety of accessory items. trol of Pseudomonas is particularly important in critical The broad antimicrobial spectrum of the composi products such as opthalmic preparations. Ps, aeruginosa, tions coupled with ready adaptability to the use of heat, which few antimicrobials control reliably, is considered and compatibility with water carriers and mixtures with by some authorities to be the most troublesome as well higher solvents such as propylene glycol, provides as the most ubiquitious. unique high level disinfection of critical items. Items of Skin odor control. The compositions of Section 1 lesser criticality are disinfected rapidly in the cold. offer unique performance in this area. Skin odor, bacte Dental antisepsis and disinfection. Hot and cold disin rial in origin, generally is caused by resident Gram-posi fection and sterilization of instruments and a broad 10 tive skin flora picked up in sweat containing nutrients range of other items is practical with the compositions for rapid bacterial growth. Presently available antimi of Section 1. For scrub-up of oral surgery personnel the crobials in underarm and foot deodorants, medicated just preceding procedures are highly effective. The soaps and the like, are relatively effective against these compositions of Section 1 exhibit particularly useful offensive odor-producing organisms, but generally are efficacy for the frequent daily hand washings required 15 ineffective against a variety of other organisms that, of dental personnel between patients and for routine while not necessarily causing disagreeable odor, are procedures. dangerous to man, especially if given the earlier de The oral cavity harbors large microbial polulations scribed opportunity to proliferate in the relative ab which are most difficult to control. Limited and general sence of Gram-positive bacteria. This is especially true oral antisepsis, particularly for prevention of post-treat 20 of the Gram-negative Pseudomonas and to a lesser ex ment infections, is effectively accomplished with the tent Candida albicans, Present antimicrobials capable of compositions of Section 1. overall broad spectrum control are not suitable for wide Disease control. The broad spectrum antimicrobial consumer use for a number of reasons including toxic activity of the compositions of Section 1 make these ity, odor, color, skin sensivity or irritation. Indeed many uniquely suitable for a wide range of pathogenic condi 25 have been banned by governmental agencies except for tions including mastitis in lactating females, bacterial perscription use. and fungal hemorrhoidal conditions; ringworm; vulvo The compositions of Section 1 have a prerequisite vaginal candidiasis; athlete's foot; oral moniliasis; my safety and other features needed for broad consumer cotic skin infections with casual agents that include usage. These compositions uniquely provide a balance Candida albican, Trichophyton sp., Epidermaphyton 30 of antimicrobial activity that prevents overgrowth of floccosum, Microsporum canis and Microsporum audo uncontrolled organisms. Entirely unexpectedly, per uini; Trichomonal and non-specific vaginitis, urinary haps due to their substantive nature, the compositions infections; hair and scalp conditions; a variety of exhibit skin odor control for much longer periods than nemotodal and tremotodal conditions. present products. Contraception. The compositions of this invention 35 Even more unexpected is the long term odor protec are spermicidal. tion provided by residual action only. When the formu Specialty water disinfection. The compositions of lated compositions are applied with water, and dried, no Section 1 find utility in therapeutic baths and various odor will develop for much longer periods than present water soaking treatments. underarm deoderants allow. Environmental control of fungal allergies. A substan Medicated, deoderant soaps are in wide use for bath tial portion of human allergic conditions is caused by a ing. Used regularly, these provide a significant degree wide range of molds and mildews which infest homes, of body odor control, although the magnitude of con living quarters and other habitated buildings. Such al trol is considerably less than that afforded by underarm lergies can be alleviated by control of the fungal organ deodorants. Further the more effective medicants, such isms. To be effective, antifungal application must be 45 as hexachlorophene, have been banned for general use. made to all surfaces and articles including walls, floors, Bathing with formulated compositions of Section 1 ceilings, crawl spaces and basements, furniture, drapes provide cleaning power equal to the best of current and carpets, mattresses and pillows. products and, quite remarkably, provide sole control of The compositions of Section 1 have the necessary body and skin odor for much longer periods of time prerequisites for such application, e.g., no color or odor, 50 than present medicated soaps for underarm and foot low taste, and no damage to or unacceptable residue on deodorant preparations, medicated vaginal douches, treated surfaces: These requirements are combined with and medicated hair shampoos. extermely high fungistatic and fungicidal efficacy. Such It would appear that the after-applicaton water insol antifungal treatment can be coupled in a number of ubility of the compositions and high sutstantivity to skin cases with cleaning, leaving a substantive and persistent 55 has a significant role in the demonstrated long term residue on carpets, upholstery, walls, floors, tiles and odor control. The insolubility/substantivity factor is other surfaces. much greater than might be expected in view of the Pharmaceutical, toiletry and cosmetic product pres extreme severity of the service conditions. ervation. The compositions of Section 1 possess near General bathing with formulated compositions of the ideal properties for these uses, including very high mi invention also provides control of the ubiquitious condi crobiological activity against the representative organ tions known as athlete's foot, jock itch, vaginitis and isms that cause product deterioration and disease to the other assorted skin and mucuous membrane conditions. user such as Staphylococcus aureus, Escherichia coli, Can Our discoveries are illustrated by the following com dida albicans, Pseudomonas aeruginosa and Aspergillis positions: niger spores. Control of the latter generally is difficult to 65 achieve. Control of Ps, aeruginosa also is difficult and of Example 1 considerable importance because this organism causes The following composition was prepared and tested pathogenic conditions in man and animals, against for skin substantivity: 4,766,113 45 46 Oxine 4.1 parts by weight No underarm irritation was noted by any test subject, Copper hydrate 1.4 parts by weight although one test subject had a history of irritation DDBSA 65.0 parts by weight problems with a number of brand name products. Propylene glycol 29.5 parts by weight Ten parts by weight of this composition was blended 5 Example 4 (heated at 140 F. with constant agitation) with: The second formulation in the preceding Example Oleyl alcohol ethoxylate (20 EO) 15 parts by weight was applied once daily by eight test subjects to the skin Lauryl alcohol ethoxylate (25EO) 25 parts by weight between the toes for control of "Athlete's Foot'. The Octyl phenol ethoxylate (9 EO) 5 parts by weight condition was brought largely under control and defi Acetylated lanolin 2 parts by weight 10 nite signs of skin healing were exhibited in all cases by Distilled water 43 parts by weight the end of 24 hours. Two of the cases had previously The resulting formulation was a transparent pale resisted control with several over-the-counter brand yellow-green, medium viscosity liquid. One gram was name antifungals. One of the two, a virulent infection, used to wash the hands. After water rinsing and drying had reached the bleeding stage from deeply cracked the hands, a drop of 1% dithio oxamide copper indica 15 skin and a bacterial infection started, despite twice daily tor was applied to various parts of the hands. Immediate application for eight days of a 1% solution of Toluaftate development of a black color indicated presence of which has gained broad recognition for effective con copper (in Cu-8-Q) on the skin. Using Ivory soap, the trol of "Athlete's Foot'. The Toluaftate product not hands were washed, rinsed and dried twice more. The only failed to control the case of "Athlete's Foot' but black color remained, requiring several days to disap 20 also the ensuing bacterial infection. The test formula pear. tion brought both the fungal and bacterial infections This formulation was an effective hand cleaner, ex under control in 24 hours. hibited no skin irritation and left the skin with a residual The causal organisms of this last fungal infection soft, smooth feel. Used for bathing and as a hair sham almost certainly were not those defined medically as 25 causing "Athlete's Foot', but rather one or more other poo, the formulation exhibited the same results. fungi which to the layman (but not a dermatologist) Example 2 produces similar or identical symptoms and hence is called "Athlete's Foot'. The Tolunaftate failed in that The final formulation of the preceding Example was its narrow-spectrum antifungal activity had no effect used to wash the entire body, including the scalp and 30 upon the causal organisms in the test case and because it hair. No skin odor developed until the middle of the has no significant antibacterial efficacy. Very few "Ath fourth day. Commercially available underarm deodor lete's Foot' preparations provide even a low order of ants (two brands) previously tested by three subjects, antibacterial activity. following a bath with a commonly used medicated bar The compositions of the invention, therefore, offer soap, failed to stop odor development for even 24 hours. 35 not only potent control of true "Athlete's Foot' but Example 3 also of a wide range of fungal and sometimes resulting The following composition was prepared and evalu bacterial conditions which to the public is "Athlete's ated for underarm odor control: Foot'. The importance of the major improvement over Oxine 2.05 parts by weight existing technology is the fact that in the vast majority Copper gydrate 0.70 parts by weight of cases the public, not a dermatologist, makes the diag DDBSA 40.00 parts by weight nosis and determines the treatment. Propylene glycol 57.25 parts by weight Example 5 Three parts of this composition was blended as before Hemorrhoids, varicose veins of the rectum, are suf with: 45 fered by an estimated 40% to 50% of the adult popula Prophlene glycol 5 parts by weight tion. Prescriptions and over-the-counter preparations, Glycerin 5 parts by weight ointments and suppositories, are in wide, almost univer Ethanol 5 parts by weight sal use for alleviation of these widespread symptoms of Water 82 parts by weight hemorrhoids: (1) bleeding from the rectum, (2) bacterial Four test subjects applied this final formulation under 50 infection which can cause inflammation of tissues and one arm and a brand name underarm deodorant under pain and (3) fungal infection (caused by organisms re the other arm. Total odor control ranged from a mini lated to "Athlete's Foot' and ringworm causal agents) mum of 3 days to almost 6 days for the test subjects which brings about itching which at times can be near whereas none of the three brand name comparative intolerable. control deodorants provided complete odor control for 55 Present state-of-the-art hemorrhoidal preparations a full 24 hours. contain an antimicrobial in a soothing, emollient oint Essentially the same odor control results were ob ment base; some also contain a topical anesthetic for tained using the following formulation in ointment pain control. The antimicrobial is designed to serve a form: dual purpose: (a) to allow healing, thereby controlling Two parts of the foregoing composition were 60 rectal bleeding, bacterial infection and pain and (b) to blended with: control itching. These objectives are difficult ones to Oleyl alcohol ethoxylate (20 EO) 18 achieve because of present unavailability of sufficiently Lauryl alcohol ethoxylate (25 mol) 9 potent yet safe antimicrobials, and because of the great Acetylated lanolin 1 difficulty of obtaining effective antimicrobial action in Glycerin 9 65 the inevitable presence of large amounts of organic Propylene glycol 4 matter. Polyvinyl pyrrolidone, 60% in water 1 The first formulation of the preceding Example was Distilled water 56 applied for hemorrhoid symptom control. Rapid and 4,766,113 47 48 more effective relief of symptoms resulted, as compared to commercially available products. In one instance -continued involving almost daily rectal bleeding for over six years Minimum Fungicidal Concentration despite daily use of a brand name product, the test for (ppm of active ingredient) mulation brought about complete cessation of bleeding A B C D within 24 hours for almost three months with no further (ATCC 13963) Aureobasidium pullulans 1. 100 - - application of the test formulation. At first resumption (ATCC 16624) of bleeding, application of the test formulation again Lenzites trabea 100 -3 - controlled all bleeding and weekly and bi-weekly appli (ATCC 11539) O Polyporus tulipiferae 100 1-3 - cation thereafter for the next six months prevented all (ATCC 11245) further bleeding. Penicilium brevi 1 100 - -- The broad-spectrum efficacy of the compositions of compactum (ATCC 16024) the invention as demonstrated by the in-vivo and in Rhizopus stolonifer 1. 100 1-3 - vitro data herein, and in my copending application re (ATCC 24794) Trichodernia viride 0 100 - - ferred to above, provide results of a magnitude not 15 (ATCC 8678) heretofore available for treatment of pathogenic condi Trichoderma sp. 100 - - tions of man and animals, and for health related applica (ATCC 12668) tions. The ideal antimicrobial is highly and rapidly toxic Candida albicans 1 1,000 -- 3 to the target organisms and non-toxic to the host organ (ATCC 10259) isms and possesses to a high degree the desirable prere 20 quisites of low taste, odor, high stability and the like. These results illustrate the high efficacy of the com The compositions of the invention come closer to the position of this invention and confirm the fact that an theoretical ideal than anything heretofore known by a antifungal composition prepared from a Cu-8- margin ranging from significantly better to almost infi Q/DDBSA solution is much superior to DDBSA alone. nitely superior. 25 The results also indicate the favorable relative efficacy of the test composition compared to the two commer Example 6 cially available fungicides of recognized high perfor The following composition was tested for minimum alC. fungicidal concentration and compared to two well known antifungal chemicals-pentachlorophenol and 30 Example 7 2,3,5trichloro-4-propyl-sulfonyl pyridine-and Using the standard AOAC fungicidal test method DDBSA. (12th Edition, 1975), the composition below was evalu Oxine 8.2 parts by weight ated against two widespread fungi. Copper hydrate 2.8 parts by weight Oxine 2.08 parts by weight DDBSA 59.0 parts by weight 35 Copper hydrate 0.70 parts by weight Propylene glycol methyl ether 30.0 parts by weight DDBSA 40.00 parts by weight All of the test composition concentrations to be Isopropanol 32.00 parts by weight tested were incorporated in the fungal growth media Demineralized water 25.33 parts by weight (agar) in accordance with standard microbiological The two fungi were Aspergillis niger, a ubiquitous practices. Agar plugs containing the test fungicides then black fungus which flourishes on a broad range of sub were inoculated with a sporulating culture and inocu strates, and Trichophyton mentagrophytes, a cause of lated at the temperatures and times specified by The 'athlete's foot.' American Type Culture Collection (ATCC) recom A. niger-at 1:200 use dilution in a water carrier, no mendations. The plugs were then scored for absence or growth after 10 minutes' exposure. presence or organism growth. The results are shown in 45 T. mentagrophytes-at 1:750 use dilution in a water the following table. Minimum fungicidal concentrations carrier, no growth after 10 minutes' exposure. were determined against a broad spectrum of fungi that Simular but somewhat lower efficacy results were are detrimental to man, foodstuffs and materials and obtained by substituting zinc-8-Q or aluminum-8-Q in which can result in metabolite formations (mycotoxins) the composition of this example, produced by reacting of extreme toxicity to man and animals. 50 zinc oxide and aluminum hydroxide respectively with In the table below, Composition A is that of this OX11e. Example 25 and the active ingredient is Cu-8-Q; Com position B is DDBSA; Composition C is pentachloro Example 8 phenol; and Composition D is 2,3,5-trichloro-4-propyl The DDBSA/Cu-8-Q solution of Example 7 diluted sulfonyl pyridine. Where no concentration is listed, no 55 1:400 in a water carrier, was applied by spray nine data is available. The stated value of "1' means 1 or times, at two-week intervals, to peach and nectarine fewer ppm. cultivars during the growing season. The results against brown rot (Monolinia fructocola), compared to non treated trees, is presented below. Minimum Fungicidal Concentration 60 (ppm of active ingredient) A. B C D % Fruit Affected Aspergilis niger l 10,000 1-3 - Peach Nectarine (ATCC 9642) Aspergillis terreus 1. 00 --- 36 At harvest (treated) 1 3 (ATCC 10609) 65 At harvest (untreated) 15 43 Aspergilis flavus 1. 1,000 22-54 - Five days later (treated) 3 6 (ATCC 11655) Five days later (untreated) 60 67 Alternaria alternata 1 4,766,113 49 50 available antibiotics, notably Pseudomonas sp. typified Example 9 by Pseudomonas aeruginosa PRD-10, the standard strain The DDBSA/Cu-8-Q solution of Example 7 was in the United States for evaluation of antibacterials for tested in vitro against a major turf pathogen, Helmin mandatory Gram-negative control applications. thosporium vagans, via a standard agar plate culture 5 The compositions herein disclosed eliminate or sub technique, with these results: stantially reduce these use limitations inherent in many other germicides. The compositions are quite unique in having strong Gram-positive and Gram-negative activ Fungus Colony Diameter ity combined with broad versatility of formulation and 1:6700 use dilution in water carrier 1 mm use plus a high degree of safety (low toxicity and zero to 1:3350 use dilution in water carrier 0 low skin and eye irritation). Add to this the high effi Control 21 mm cacy, broad spectrum antifungal activity of the compo sitions of this invention and the resulting range of toxic The results demonstrate very high efficacy in con ity to target organisms and safety to man, the most trolling this important pathogen. Complete control of 15 sensitive of hosts, is unique indeed. H. vagans was achieved in this assay between 3.7 and 7.5 The balance of toxicity provided by this invention to ppm of Cu-8-Q. Gram-positive, Gram-negative and fungal microorgan As illustrated in the examples to follow, the composi isms has special value in the broad consumer field of tions of this invention have high efficacy against a broad skin deodorancy. Present antibacterials suffer from the spectrum of bacteria that are pathogenic to mammals 20 fact that they are effective primarily against Gram-posi and plant life, that contribute to reduced water quality, tives, allowing Gram-negative and fungi overgrowth, a that cause deterioration of foodstuffs, that degrade a condition considered dangerous by many authorities. broad range of manufactured and natural materials and products, and which generate toxic metabolites (bac Example 10 teria-toxins) that are among the most poisonous sub- 25 The DDBSA/Cu-8-Q solution of Example 6 was stances known to man. tested, along with a number of well known antimicro Of particular interest in high efficiency against Gram bial agents, against abroad spectrum screen of economi negative as well as Gram-positive microorganisms. Few cally important Gram-positive and Gram-negative bac antibacterial materials now available are effective teria. All antimicrobial agents were incorporated in the against the Gram-negatives and still fewer provide eco- 30 agar bacterial growth media according to standard mi nomical control of them. A number of available antibac crobiological practices. The bacterial species were terials toxic to Gram-negative organisms have practical grown in nutrient broth; 24-hour cultures, the inocu limitations which severely restrict use, including high lum, then were streaked onto the nutrient agar plates mammalian toxicity, phytotoxicity, corrosiveness to containing the test antimicrobials. After a 24-hour incu skin and a variety of materials, strong odor, strong 35 bation at the appropriate temperature, the plates were color, high volatility, low or erratic shelf stability, low rated for presence or absence of bacterial growth. or nonexistent residual activity, and prohibition of use Minimum bactericidal concentrations for each of the at elevated temperatures. tested agents are stated in the following tabulation of The basic significance in the need for Gram-negative results in parts per million (ppm) of active ingredient as control lies in the fact that this bacterial category in- 40 defined in the description of each agent. cludes a number of widespread, virulent pathogens which are difficult to impossible to control with present

Antimicrobial Agent (ppm of active ingredient) Bacteria (ATCC No.) A B C D E F G Gram-positive: Bacillus cereus 1 100 - 7 8 5-10 m Bacillus lichenforms (27326) 100 --- 7 8 2-5 r Bacillus megaterium (27327) 100 -- 7 8 ------Bacillus subtilis (37328) 100 - 750 8 --- 3 Micrococcus flavus (10240) l 100 ww. 7 8 - m Mycobacteriumphlei (15610) 1 10 - 7 8 - 3 Staphylococcus aureus (6538) 1. 100 2083 7 8 1-3 3

Alcaligenes faecalis (337) 10 1000 - 750 80 --- m Alcaligenes marshali (21030) 04 100 - 7 8 - m Esherichia coli (11229) 104 10,000 -- 750 80 250-500 65 Flavobacterium arboresceus (4358) 10 10 466 7 8 --- m Klebsiella pneumoniae (4356) 10 10,000 - 750 8 - -- Proteus vulgaris 10 1000 --- 750 800 - m Pseudomonas aeruginosa (15442) 104 1000 466 750 800 1000-2500 165 Salmonella cholerasuis (10708) 104 1000 - 750 80 250-500 165 Salmonella typhi (6539) 104 1000 2083 750 80 - 16S "stated value of "1" means l or less A = the DDBSA/Cu-8-Q solution of Example 6 with the active ingredient expressed in terms of Cu-8-Q. B = DDBSA. C = phenol D = Betadine, an iodine/polyvinylpyrrollidone complex containing 0.75% iodine. The active ingredient is iodine. E = Alkyl dimethyl ammonium chlorides (61% C, 23% C4, 11% C6 and 3%. Co). F = sodium pentachlorophenate G = 2,3,5-trichloro-4-propylsulfonyl pyridine. 4,766,113 51 52 These data demonstrate the high efficacy of the com result may be infected potato plants, tubers and reduced position of Example 6. On the basis of the average of the yields. efficacies against all the test organisms, Composition A The test procedure consisted of dipping unpainted, is 45 times superior to Composition B; 88 times better planed wood laths (6') into a slurry of infected ring rot than Composition C; 10.7 times better than Composition 5 tuber tissue, allowing excess slurry to drain off (3-5 D; and 3.5 times superior to Composition E. minutes) and then spraying the contaminated lath with On the basis of the average of the efficacies against the test antibacterials. Three to five minutes later, the three test bacteria (Staphylococcus aureus, Salmo healthy Norgold Russet potato seed pieces were rubbed nella cholerasuis and Pseudomonas aeruginosa PRD-10) vigorously against both sides of the contaminated and required by the Environmental Protection Agency of a O antibacterial-treated laths. The process was repeated “hospital grade' disinfectant, Composition A is 10 times using laths not contaminated with C. Sepedonicum but better than Composition B; 7.2 times better than Com treated with the test antibacterial agent. The rubbed position D; and 1.6 times better than Composition E. seed pieces were stored in bags and later planted at the appropriate time. Example 10A 15 In addition to the composition of the invention, un The composition set forth below was prepared by treated controls, 20% Clorox (1.05% sodium hypochlo previously described procedures: rite in water), formaldehyde (37% formalin diluted Oxine 2.08 parts by weight 1:120 in water) and Roccal (benzalkonium chloride or Copper hydrate 0.70 parts by weight zephiran chloride) diluted with water to 800 ppm con Iospropanol 32.00 parts by weight 20 centration were tested. The results of the test are tabu DDBSA 40.00 parts by weight lated below and refer to plants and tubers produced Water (demineralized) 25.22 parts by weight from the tubbed test seed pieces.

Antimicrobial Ring Rot Contaminated 2% Plant Stored % Ring Rot Plants % Ring Rot Tubers Yield cwt/acre None (control) Yes 98 23 8 493 None (control) No 95 O O 609 DDBSA/Cu-8-Q Yes 98 O 2 631 DDBSA/Cu-8-Q No 100 O 0 602 20% Clorox Yes 98 20 9 500 20% Clorox No 98 O 0 602 Roccal Yes 98 20 9 515 Roccal No 98 0 0. 638 The composition of this example demonstrates supe When evaluated as a bactericide by the AOAC Use 35 rior control of the ring rot bacterium. Other species of Dilution Method (12th Edition, 1975), 10 ring carriers the genus Corynebacterium are causal agents of disease per organism, the following results were obtained (A=- in man and a variety of plant life. subculture and B=resubculture): Example 12 The composition below was prepared and tested for Use Dilution in Negative Positive 40 speed and range of antibacterial activity, in the absence Water Carrier A. B A B and presence of organic matter (blood) for use in hospi Staphylococcus aureus 1:1000 10 10 O O tal disinfection, cold sterilization and antisepsis. Salmonella cholerasuis 1:1000 10 10 O O Oxine 4.1 parts by weight (PRD-10) Copper hydrate 1.4 parts by weight Pseudomonas aeruginosa 1:400 10 10 0 0 45 DDBSA 65.0 parts by weight Aerobacter aerogenes 1:400 10 10 0 O Propylene glycol methyl ether 29.5 parts by weight Many antimicrobial agents are partially or totally A ten-minute kill is required against the first three deactivated in the presence of organic matter, constitut pathogens for sale as a hospital grade disinfectant. Effi ing a severe limitation to effectiveness of such agents for cacy against the fourth organism, a major cause of slime 50 a number of uses such as wound antisepsis and medical to recirculated cooling water systems and pulp and instrumentation and surface disinfection where large paper mills, demonstrates utility of the composition as a amounts of organic matter often are encountered and slimicide. sometimes are unavoidable. The AOAC Use Dilution Confirmation Test (12th Example 11 55 Edition, 1970) was modified as follows: This composition was prepared and tested at one use (a) The test temperature was 37 C. dilution, 1:50 in water carrier, against the causal agent (b) The ring carriers were soaked in sheep blood for of potato ring rot bacteria (Corynebacterium sepedoni two hours, air-dried for one hour, then contami cum): nated with the test pathogen. Oxine 2.08 parts by weight 60 (c) The contaminated rings were contacted with the Copper hydrate 0.70 parts by weight test antibacterial agent for 30-second, one-minute DDBSA 40.00 parts by weight and three-minute periods. Triton X-100 20.00 parts by weight The results are set forth in the following table, in Isopropanol 22.00 parts by weight which: Water (demineralized) 15.22 parts by weight 65 0=no growth in 10 of 10 tubes tested Infected potato seed readily contaminate potato seed 1=growth in 1 of 10 tubes tested cutters, sacks, bins, cellars, trucks and planting equip 2=growth in 2 of 10 tubes tested ment with the highly infectious ring rot bacteria. The 3 = growth in 3 of 10 tubes tested etc. 4,766,113 53 54

Use Dilution in In Absence of Blood In Presence of Blood Test Pathogen Water Carrier 30 sec. 1 min. 3 min. 30 sec. min. 3 min. Staphylococcus aureus Undiluted O O O O O O (ATCC 6538) 1:10 O O O O 0 O 1:100 O 0 O 3 6 3 Salmonella typhi Undiluted O O O O O O (ATCC 6539) 1:10 O O O O 0. O 1:00 O O 0 O 0 0 Pseudomonas aeruginosa Undiluted O O 0 O O O (ATCC 15442) 1:10 O O 0 10 10 O 1:100 0 O 0 10 3 3

The above composition was prepared by the proce 15 dure already described and evaluated for efficacy These results indicate that the test composition is against ciliated protozoan and two types of viruses. capable of rapid antibacterial action in the presence of substantial amounts of organic matter against the three human pathogens generally considered as definitive for Hemaglutination Assays antibacterial efficacy evaluation. 20 HA Titre Virus Untreated Treated (1:50)" Treated (1:200) Example 13 Adenovirus 128 0 0 Newcastle 512 O 0 The composition of Example 7 was prepared for Disease virus evaluation as a skin degerming agent against resident (1:50 and 1:200 use dilutions of composition in a water carrier in contact with virus and transient flora. Six subjects were tested using the 25 suspensions for 15 minutes) Modified Prices Multiple Basin Technique which mea sures reduction of skin flora as a percentage of that The HA titre is a measure of the number of infectious achieved by washing the hands with unmedicated soap. virus particles present in the test suspension. Bacterial counts were taken from the first, fourth and fifth basins. The counts from the first basin represent the 30 Plaque Assays transient bacterial flora and that of the fourth and fifth Via the same procedure as above, the untreated virus basins the resident flora. Prior to the test, none of the suspensions contained 6.4x 10 pfu/ml of Adenovirus subjects used a medicated soap for one week. virus particles and 21 x 105 pfu/ml Newcastle Disease The results set forth in the following table demon virus particles respectively. After treatment with the strate a very high order of efficacy for the test composi 35 1:50 and 1:200 use dilutions, readings of 0 pfu/ml were tion for critical degerming uses such as a surgical scrub obtained. Each pfu represents one infectious virus parti for operating room personnel and skin preparation at cle. A zero pfu reading represents total inactivation of the surgical site. the infectious virus. Protozoan Inhibition Unmedicated Soap Test Composition % 40 Average No. of Average No. of Reduc Inhibition of growth of ciliated protozoan (Tetrahy Subject Basin Organisms/Basin Organisms/Basin tion mena) in pond water was obtained at a 6 ppm concen 1 l 2,500,000 60,000 97.6 tration (based on Cu-8-Q) of the above test composition 4. 1,900,000 25,000 98.7 of DDBSA and Cu-8-Q after 6-hour and 72-hour 5 1,300,000 15,000 98.8 45 contact times. The 6 ppm reading represents the MIC 2 1 1,700,000 30,000 98.2 4. 1,100,000 20,000 98.2 (Minimum Inhibitory Concentration). These results 5 800,000 1,000 99.9 demonstrate high efficacy of the test composition 3 1. 800,000 40,000 95.0 against the test microorganisms. 4. 600,000 20,000 96.7 5 500,000 1,000 99.8 50 SECTION 7 4. l 2,000,000 50,000 97.5 4. 1,000,000 1,000 99.9 We have also discovered than the compositions of 5 800,000 1,000 99.9 Section 1 are useful for controlling plant disease by 5 l 1,500,000 40,000 97.3 contacting plants with such compositions. More partic 4 800,000 1,000 99.9 5 500,000 1,000 99.8 ularly, this embodiment of our invention relates to the 6 l 1900,000 50,000 97. 55 control of plant disease and more particularly to the use 4. 800,000 15,000 98.1 of certain antimicrobial agents that are comparatively 5 520,000 1,000 99.8 nontoxic to animal and plant life but are highly effica ceous against fungal, bacterial and other classes of pathogens, including viruses and nematodes, which Example 14 60 microorganisms are important casual agents of disease Oxine 2.08 in a broad spectrum of plant life. Copper hydrate 0.70 We have found that the compositions of section 1 Nonyl phenol/EO surfactant 20.00 exhibit a wide range of utility against fungal, bacterial DDBSA 45.00 65 and other classes of pathogenes, including viruses and Isopropanol 15.00 nematodes, which are important casual agents of dis Water (distilled) 17.22 eases in a broad spectrum of plant life. Such plant life (1 molnonyl phenol to 6 mols ethylene oxide) includes field and horticultural crops, ornamentals and grasses. The mode of use includes foliar, seed, dormant 4,766,113 55 56 season and, in some instances soil treatment. It also reduces the initially low mammalian toxicity, as applied includes treatments designed to preserve the plant prod to the host plant or soil, up to about 95% in a few weeks uct in the post-harvest period, which treatment may be after application without a corresponding reduction in carried out both pre- and post-harvest. residual antimicrobial efficacy. Second, death in the The disclosed compositions present an unusual com- 5 event of accidental ingestion of a lethal dose of the bination of properties that are unexpectedly unique for compositions is a secondary and reversible mechanism control of a broad range of diseases of a large number of wherein the time period for initiation of medical treat plant species, including: ment is non-critical. Medical treatments for this condi Antifungal control. The spectrum of activity in the tion are well-known and are generally successful with use of compositions of this invention covers the four 10 no adverse after effects. classes of fungi-Ascomycetes, Basidiomycetes, Phyco High uniformity of coverage. Appropriate use-dilu mycetes and Fungi Imperfecti tions of the compositions of this invention exhibit pro Antibacterial control. The use of compositions of this nounced substrate wetting and spreading properties invention covers a broad spectrum of Gram-positive which provides a high degree of uniformity of coverage and Gram-negative plant pathogens. 15 of the host plant. Antiviral and Antinematodal control. The use of Ease and economny of handling and application. The compositions of this invention is effective against vi compositions of this invention generally are concen ruses and a substantial selection of nematode species. trated, readily pourable liquids which are easily and High antimicrobial efficacy. Minimum inhibition and rapidly misible with water for use to form either true minimum kill concentrations of the compositions of this 20 solutions or colloidal emulsions, as diluted for use, they invention are very low. are easy to apply with no settling out of active ingredi Low phytotoxicity. Generally, but not always, the ents. Due to the high product concentration, a mini use of the compounds of this invention have low phyto mum of product need be transported, handled and toxicity at adequate use. stored and disposal of containers is minimized. Water solubility. Concentrates prepared for use in 25 In the plant world, bacterial diseases are in the minor this invention may be use-diluted in a variety of polar ity relative to fungal diseases. Surprisingly, very few and non-polar carriers, including water which is the products are available for bacterial disease control. This most versatile. Diluted with water, the concentrates is in sharp contrast with the very large availability of generally form true solutions that in most cases provide products for fungal diseases. The antibacterials now substantial handling, mixing-for-use, application and 30 available have quite limited utility because of either low uniformity-of-coverage advantages over the typical efficacy or high cost. Compositions of the invention water-insoluble products. In some cases of use-dilution, exhibit considerable antibacterial promise for plant use stable and often colloidal emulsions are formed. These by reason of high unit efficacy and low cost. generally provide the same advantages as true solutions. The method of use and efficacy of the compositions Water insolubility. On application to the host organ- 35 of this invention are illustrated in the following exam ism (or the soil), the compositions of this invention turn ples. insoluble in water in most instances, which increases antimicrobial persistence of activity and results in in EXAMPLES creased rain wash off resistance. Example 1 Substantivity. When applied to a number of sub- 40 strates, the compositions of this invention become The following composition was prepared and tested tightly affixed (substantive) and consequently exhibit as a rice seed treatment: substantial resistance to removal by rain or other fac Oxine 2.05 parts by weight tors. Copper hydrate 0.70 parts by weight Heat and sunlight stability. The composition of this 45 DDBSA 40.00 parts by weight invention exhibit high stability to heat (to over 200 F) Octyl phenol ethoxylate (9EO) 20.00 parts by weight and light, particularly ultraviolet. Isoporpanol 20.00 parts by weight Safety. Acute oral mammalian toxicity of the highest Water 17.25 parts by weight practical concentrations of the compositions of this This composition was placed in water (800 cc per invention is moderately low, about halfway between 50 cwt. of seed) and used to coat the inside walls of a aspirin and common table salt. Typical use dilutions container. Weighed seed samples were placed in the thereof, 1:100 to 1:1000 in water, are nontoxic. container which was rotated for 15 minutes to transfer Skin and eye irritation properties of typical use-dilu the test composition to the seed. The treated seed was tions range from very low to zero. Highest practical stored until planted. This application procedure was concentrates of the compositions exhibit moderately 55 repeated for application of several commercially avail high skin and eye irritation. At one-tenth of maximum able antifungal products. concentration, use-dilutable in the 1:10 to 1:100 range, Treated and untreated control seed was planted in the compositions are inocuous to the skin and show low thirteen locations in four states using both water-plant eye irritation. ing and drill-planting methods. The effectiveness of all The toxicity characteristics of the compositions ex- 60 tested products in increasing the seedling emergence hibit two unusual and unique features of considerable rate is shown below as a consolidation of results from all practical value. First, biodegradation in most instances test plots.

(1) (2) (3) (4) Application Improvement Performance Range of Improvement (%) Rate over Control (%) Consistency High Low EXAMPLE 2 4. 18.3 36.2 plus 38 plus 2 4,766,113 -continued (1) (2) (3) (4) Application Improvement Performance - Range of Improvement (%) Rate over Control (%). Consistency High Low PRODUCTA 4 27.4 80.4 plus 56 minus 2 PRODUCT B 4. 16.6 63.1 plus 61 minus 2 PRODUCT C 4. 12.6 47.8 plus 32 minus 16 PRODUCTD 4. 12.3 71.3 plus 37 minus 35 (1) Ounces per 100 lbs. (cwt.) of rice seed of the chemical active ingredient in all instances except Example 2 which is given in terms of the total composition. "contains 0.1 ounce of Cu-8-Q. (2) Percentage increase in seedling emergence rate as compared to that of the untreated control seed in the thirteen test plots. (3) These numbers express the difference between the highest and lowest precent change in seedling emergence rate for each test product compared to the untreated seed results in the thriteen test plots. Thus, the lower the number, the greater is thetest-to-test consistency of emergence rate improvement provided by the test product. (4) This column lists the best (high) of the thirteen test results and the poorest (low). "Plus" represents improvement over untreated control seed emergence rate whereas "minus" represents a reduction therein. Product A - a coordination product of zinc ion and manganese ethylene bisdithiocarbamate. Product B - copper hydroxide formulated as a finely divided flowable composition. Product C - cis-N-(1,1,2,2-tetrachloroethylthio)-4-cyclohexene-1,2-dicarboximide. Product D - 5,6-dihydro-2-methyl-1,4-oxathiim-3-carboxanilide. The estimated costs of Products A through Drange from 3 to 5 times higher than that of the composition of Example 2.

Example 2 -continued The following composition was tested for minimum Minimum Fungicidal Concentration fungicidal concentration and compared to two well 25 (ppm of active ingredient) known antifungal chemicals-pentachlorophenol and A. B C D 2,3,5 trichloro-4-propyl-sulfonyl pyridine-and (ATCC 11245) DDBSA. Penicilium brevi 100 - - compactum (ATCC 16024) Oxine 8.2 parts by weight Rhizopus stolonifer 1. 100 1-3 - Copper hydrate 2.8 parts by weight 30 (ATCC 24794) DDBSA 59.0 parts by weight Trichoderna viride 10 100 - - Propylene glycol methyl ether 300 parts by weight (ATCC 8678) Trichoderma sp. 1 100 - - All of the test composition concentrations to be (ATCC 12668) tested were incorporated in the fungal growth media Candida albicans l 1,000 - 3 (agar) in accordance with standard microbiological 35 (ATCC 10259) practices. Agar plugs containing the test fungicides then were inoculated with a sporulating culture and inocu These results illustrate the high efficacy of the com lated at the temperatures and times specified by The position of this invention and confirm the fact that an American Type Culture Collection (ATCC) recom antifungal composition prepared from a Cu-8- mendations. The plugs were then scored for absence or 40 Q/DDBSA solution is much superior to DDBSA alone. presence of organism growth. The results are shown in The results also indicate the favorable relative efficacy the following table. Minimum fungicidal concentrations of the test composition compared to the two commer were determined against a broad spectrum of fungi that cially available fungicides of recognized high perfor are detrimental to man, foodstuffs and materials and which can result in metabolite formations (mycotoxins) 45 face. of extreme toxicity to man and animals. Example 3 In the table below, Composition A is that of this The compositions of this invention exhibit efficacy Example 2 and the active ingredient is Cu-8-Q; Compo against a broad spectrum fungal plant pathogens, as sition B is DDBSA; Composition C is pentachlorophe illustrated by various use dilutions in a water carrier of nol; and Composition D is 2,3,5-trichloro-4-propylsul- 50 the following composition: fonyl pyridine. Where no concentration is listed, no Copper hydrate 1.70 parts by weight data is available. The stated value of '1' means 1 or 8-hydroxy quinoline 4.44 parts by weight fewer ppm. Isopropanol 35.00 parts by weight DDBSA 58.86 parts by weight Minimum Fungicidal Concentration 55 A. Valencia Oranges (ppm of active ingredient) Tested on harvested fruit against Phomopsis stem A B C D end rot and Diplodis rot, at a 1:100 use dilution, 2 Aspergillis niger 10,000 1-3 - minute dip application. After 3 weeks at 70 F., the (ATCC 9642) following percentages of decay were noted: Aspergillis terreus 100 - 36 60 Control (untreated) oranges-9.5% decay (ATCC 10609) Aspergillis flavus 1. 1,000 22-54 - Treated oranges-5.3% decay (ATCC 11655) B. Sugar Cane Alternaria alternata ------An agar seeding test against Ceratocystis paradoxa (ATCC 13963) (pineapple disease) at a 1:10,000 (100 ppm) use dilution Aureobasidium pullulans l 100 - - 65 (ATCC 16624) yielded a 3.0 mm. inhibition zone. Lenzites trabea 1. 100 1-3 -- C. Peach Trees (ATCC 11539) Tested against Taphrina deformans (causes leaf curl Polyporus tulipiferae 1 OO 1-3 H disease). Four test trees were sprayed twice, two weeks 4,766,113 59 60 apart, with a 1:400 use dilution. Three months later, 100 (Erysiphe polygoni), bean rust (Uromyces phaseoli typica) leaves on each test tree were rated for leaf curl: and rice spot (Helminthosporium orazae and Cerocospora Control (untreated) leaves-100% leaf curl orazae). Treated leaves-13.5% leaf curl D. Cotton 5 Effectiveness against 11 fungi and 1 bacterium (Xan Bean Mildew Bean Rust Rice Spot thomonas malvacearum) that are associated with disease Example 28 composition 8 10 10 Karathane c 10 - - of cottonseed, seedlings and other plants was evaluated Vitavax - 9 - in vitro, using the following compositions: Daconil - --- 10 10 Maneb - - 8 Composition #1 Copper hydrate 1.70 parts by weight 8-hydroxy quinoline 4.44 parts by weight Methanol 4.00 parts by weight Example 5 Isopropanol 30.86 parts by weight DDBSA 59.00 parts by weight The following composition was evaluated (diluted Composition #2 Copper hydrate 2.80 parts by weight 15 with water for use) in vitro for inhibition against two 8-hydroxy quinoline 8.20 parts by weight fungal pathogens, Botrytis sp. and Alternaria sp., causal Methanol 4.00 parts by weight agents of a variety of plant diseases. Isopropanol 26.00 parts by weight Oxine 2.08 parts by weight DDBSA 59.00 parts by weight Copper hydrate 0.70 parts by weight 20 DDBSA 40.00 parts by weight Both compositions were prepared in accordance with Triton X-100 20.00 parts by weight procedures stated in previous examples. Isopropanol 22.00 parts by weight The following results were obtained, expressed in Water (demineralized) 15.22 parts by weight parts per million (ppm) of total test composition in The zone of inhibition agar plate test also was used to water carrier and the relative growth inhibition pro- 25 test Cunilate 2174 (diluted in mineral spirits for use) for vided at each test strength on each tested organism. In comparison. The composition concentrations in the the tables below: table of results below are expressed in parts per million 0=no apparent inhibition (ppm) of Cu-8-Q. The larger the inhibition zone, the 1=some inhibition greater is the efficacy of the composition. 2= considerable inhibition (little growth) 30 3=total inhibition (no growth)

Composition #1 Composition #2 Concentrations Concentrations (ppm Test Organisms O 1 5 25 100 500 1000 0 1 5 25 100 500 1000 Pythium ultimum (41 B) 0 0 0 1 2 3 3 0 0 1 2 2 2. 2 Rhizoctonia solani (1D) 0 0 0 1 2 3 3 O 0 1 2 2 2 2 Fusarium (4A) 0 0 O 3 3 3 3 0 0 1 1 2 2 Fusarium (4D) 0 1 1 2 2 2 3 0 0 1 1 2 2 2 Fusarium roseum (4C) 0 0 0 1 2 2 3 0 0 O 2 2 2 2 Colletotrichum gossypii (35A) 0 0 0 1 2 2 2 0 0 0 2 2. 2 2 Xanthomonas malvacearum (2A) 0 0 0 0 2 2 3 0 0 0 1 1. 2 2 Aspergillis sp. 1 l Helminthosporium oryzae 2 2 Mucor nucedo l Penicillium sp. Rhizopus sp. 1. 1.

Example 4 Botrytis Alternaria The following composition was screened for fungal 50 DDesAvcso pathogen response as a foliar spray on beans and rice. Concentration (ppm) 21 50 25 21 50 125 Oxine 2.08 parts by weight Inhibition zone (mm) 13 15 22 0 16 21 Copper hydrate 0.70 parts by weight Cunilate 2174 - DDBSA 40.00 parts by weight Concentration (ppm) 83 200 500 83 200 500 Isopropanol 32.00 parts by weight 55 inhibition zone (mm) 13 14 16 O 0 14 Demineralized water 25.22 parts by weight The rating scale is from 0 (no pathogen control) to 10 The DDBSA/Cu-8-Q solution of this invention ex (complete pathegon control). The concentration of hibits an improvement in efficacy against the tested active ingredient (in a water carrier) of all compositions organisms by a factor of 4x in the case of Botrytis to 10x tested is 33 parts per million (ppm). The active ingredi- 60 in the case of Alternaria. ent in the composition of Example 4 is expressed in E le 6 terms of Cu-8-Q and the chemical as listed below for xample four comparative products. The comparative products The DDBSA/Cu-8-Q solution of the preceding ex tested were Karathane (2,4-dinitro-6-(2-octyl phenyl ample (31), diluted 1:400 in a water carrier, was applied crotonate), Vitavax (5,6-dihydro-2-methyl-1,4-oxathin- 65 by spray nine times, at two-week intervals, to peach and 3-carboxanilide), Daconil (tetrachloroisophthalonitrile) nectarine cultivars during the growing season. The and Maneb (manganese ethylenebisdithiocarbamate). results against brown rot (Monolinia fructocola), com The plants and diseases tested were bean mildew pared to nontreated trees, is presented below. 4,766,113 61 62 or nonexistent residual activity, and prohibition of use at elevated temperatures. % Fruit Affected The basic significance in the need for Gram-negative Peach Nectarine control lies in the fact that this bacterial category in At harvest (treated) 3 At harvest (untreated) 15 43 cludes a number of widespread, virulent pathogens Five days later (treated) 3 6 which are difficult to impossible to control with pres Five days later (untreated) 60 67 ently available antibiotics, notably Pseudomonas sp. typified by Pseudomonas aeruginosa PRD-10, the stan dard strain in the United States for evaluation of anti Example 7 10 bacterials for mandatory Gram-negative control appli cations. The DDBSA/Cu-8-Q solution of Example 6 was The compositions herein disclosed eliminate or sub tested in vitro against a major turf pathogen, Helmin stantially reduce these use limitations inherent in many thosporium vagans, via a standard agar plate culture other germicides. The compositions are quite unique in technique, with these results: 15 having strong Gram-positive and Gram-negative activ ity combined with broad versatility of formulation and Fungus Colony Diameter use plus a high degree of safety (low toxicity and zero to 1:6700 use dilution 1 mm low skin and eye irritation). Add to this the high effi in water carrier cacy, broad spectrum antifungal activity of the compo 1:3350 use dilution 0. 20 sitions of this invention and the resulting range of toxic in water carrier ity to target organisms and safety to man, the most Control 21 mm sensitive of hosts, is unique indeed. The balance of toxicity provided by this invention to The results demonstrate very high efficacy in con Gram-positive, Gram-negative and fungal microorgan trolling this important pathogen. Complete control of 25 isms has special value in the broad consumer field of H. vagans was achieved in this assay between 3.7 and 7.5 skin deodorancy. Present antibacterials suffer from the ppm of Cu-8-Q. fact that they are effective primarily against Gram-posi As illustrated in the examples to follow, the composi tives, allowing Gram-negative and fungi overgrowth, a tions of this invention have high efficacy against abroad condition considered dangerous by many authorities. spectrum of bacteria that are pathogenic to mammals 30 and plant life, that contribute to reduced water quality, Example 8 that cause deterioration of foodstuffs, that degrade a The DDBSA/Cu-8-Q solution of Example 2 was broad range of manufactured and natural materials and tested, along with a number of well known antimicro products, and which generate toxic metabolites (bac bial agents, against a broad spectrum screen of economi teria-toxins) that are among the most poisonous sub 35 cally important Gram-positive and Gram-negative bac stances known to man. teria. All antimicrobial agents were incorporated in the Of particular interest is high efficacy against Gram agar bacterial growth media according to standard mi negative as well as Gram-positive microorganisms. Few crobiological practices. The bacterial species were antibacterial materials now available are effective grown in nutrient broth; 24-hour cultures, the inocu against the Gram-negatives and still fewer provide eco lum, then were streaked onto the nutrient agar plates nomical control of them. A number of available antibac containing the test antimicrobials. After a 24-hour incu terials toxic to Gram-negative organisms have practical bation at the appropriate temperature, the plates were limitations which severely restrict use, including high rated for presence or absence of bacterial growth. mammalian toxicity, phytotoxicity, corrosiveness to Minimum bactericidal concentrations for each of the skin and a variety of materials, strong odor, strong 45 tested agents are stated in the following tabulation of color, high volatility, low or erratic shelf stability, low results in parts per million (ppm) of active ingredient as defined in the description of each agent.

Antimicrobial Agent (ppm of active ingredient) Bacteria (ATCC No.) A. B C D E F G

Bacillus cereus 1 100 - 7 8 5-10 -- Bacillus lichenforms (27326) 1. 100 - 7 8 2-5 - Bacillus megaterium (27327) 1. 100 --- 7 8 m - Bacillus subtilis (37328) 1 100 - 750 8 m 3 Micrococcus flavus (10240) 1 100 w 7 8 m - Mycobacteriumphlei (15610) 1 10 -- 7 8 www. 3 Staphylococcus aureus (6538) 1 100 2083 7 8 1-3 3 Gram-negative: Alcaligenes faecalis (337) 10 1000 - 750 80 m - Alcaligenes marshali (21030) 104. 100 -- 7 8 - - Esherichia coli (11229) 104. 10,000 - 750 80 250-500 165 Flavobacterium arboresceus (4358) 10 10 466 7 8 m - Klebsiella pneumoniae (4356) 10 10,000 - 750 8 m --- Proteus vulgaris 10 1000 - 750 800 1m ------Pseudomonas aeruginosa (15442) 104 1000 4166 750 800 1000-2500 165 Saimonella cholerasuis (10708) 104 1000 --- 750 80 250-500 165 4,766,113 63 -continued Antimicrobial Agent (ppm of active ingredient) Bacteria (ATCC No.) A B C D E F G Salmonella typhi (6539) 104 1000 2083 750 80 165 'stated value of "1" means i or less A = the DDBSA/Cu-8-Q solution of Example 2 with the active ingredient expressed in terms of Cu-8-O. B = DidsSA. C = phenol D = Betadine, an iodine/polyvinylpyrrollidone complex containing 0.75% iodine. The active ingredient is iodine. E = Alkyl dimethyl ammonium chlorides (61% C12, 23%. C14, 1% C6 and 3% C10). F is sodium pentachlorophenate G = 2,3,5-trichloro-4-propylsulfonyl pyridine.

These data demonstrate the high efficacy of the com ment with the highly infectious ring rot bacteria. The position of Example 2. On the basis of the average of the 15 result may be infected potato plants, tubers and reduced efficacies against all the test organisms, Composition A yields. is 45 times superior to Composition B; 88 times better The test procedure consisted of dipping unpainted, than Compositions C; 10.7 times better than Composi planed wood laths (6') into a slurry of infected ring rot tion D; and 3.5 times superior to Composition E. tuber tissue, allowing excess slurry to drain off (3-5 On the basis of the average of the efficacies against 20 minutes) and then spraying the contaminated lath with the three test bacteria (Staphylococcus aureus, Salmo the test antibacterials. Three to five minutes later, nella cholerasuis and Pseudomonas aeruginosa PRD-10) healthy Norgold Russet potato seed pieces were rubbed required by the Environmental Protection Agency of a vigorously against both sides of the contaminated and "hospital grade' disinfectant, Composition A is 10 times antibacterial-treated laths. The process was repeated better than Composition B; 7.2 times better than Com 25 using laths not contaminated with C. sepedonicum but positions D; and 1.6 times better than Composition E. treated with the test antibacterial agent. The rubbed Example 9 seed pieces were stored in bags and later planted at the The composition set forth below was prepared by appropriate time. previously described procedures: In addition to the composition of the invention, un Oxine 2.08 parts by weight 30 treated controls, 20% Clorox (1.05% sodium hypochlo Copper hydrate 0.70 parts by weight rite in water), formaldehyde (37% formalin diluted Isopropanol 32.00 parts by weight 1:120 in water) and Roccal (benzalkonium chloride or DDBSA 40.00 parts by weight Zephiran chloride) diluted with water to 800 ppm con Water (demineralized) 25.22 parts by weight centration were tested. The results of the test are tabu When evaluated as a bactericide by the AOAC Use 35 lated below and refer to plants and tubers produced Dilution Method (12th Edition, 1975), 10 ring carriers from the tubbed test seed pieces. per organism, the following results were obtained (A=- subculture and B =resubculture): Ring Rot % % % Yield Contam- Plant Ring Rot Ring Rot cwt/ 40 Antimicrobial inated Stored Plants Tubers aCe Use Dilution Negative - Positive None (control) Yes 98 23 8 493 in Water Carrier A B A B None (control) No 95 O O 609 Staphylococcus aureus 1:1000 10 10 O O DDBSA/ Yes 98 O 2 631 Salmonella cholerasuis 1:1000 10 10 0 0 Cu-8-Q (PRD-10) 45 DDBSA/ No 100 O O 602 Pseudomonas aeruginosa 1:400 10 10 0 0 Cu-8-Q Aerobacter aerogenes 1:400 10 10 0 O 20% Clorox Yes 98 20 9 500 20% Clorox No 98 O O 602 Roccal Yes 98 20 9 515 A ten-minute kill is required against the first three Roccal No 98 O O 638 pathogens for sale as a hospital grade disinfectant. Effi 50 cacy against the fourth organism, a major cause of slime The composition of this example demonstrates supe in recirculated cooling water systems and pulp and rior control of the ring rot bacterium. Other species of paper mills, demonstrates utility of the composition as a the genus Corynebacterium are causal agents of disease slimicide. in man and a variety of plant life. 55 Example 10 SECTION 8 This composition was prepared and tested at one use According to another embodiment of our invention dilution, 1:50 in water carrier, against the causal agent we have found that wood can be preserved by treating of potato ring rot bacterial (Corynebacterium sepedoni the wood, using standard techniques such as dipping or cum): pressurre impregnation, with a composition as set forth Oxine 2.08 parts by weight in Section 2 (hereinafter referred to as "the composi Copper hydrate 0.70 parts by weight tions of this invention'). DDBSA 40.00 parts by weight Triton X-100 20.00 parts by weight DESCRIPTION OF THE PRIOR ART Isopropanol 22.00 parts by weight 65 Essentially, three different classes of compounds are Water (demineralized) 15.22 parts by weight currently being used for the preservation of wood. Infected potato seeds readily contaminate potato seed These include chlorinated phenols, such as pentachloro cutters, sacks, bins, cellars, trucks and planting equip phenol (PCP), mixed metal salts such as those contain 4,766,113 65 66 ing copper, chromium and arsenic (CCA salts), and depending upon the particular one used, in that they petroleum distillation by products such as creosote. sometimes are leached from the wood in service, they These preservatives may be applied to the surface of the are not as highly absorbed by the wood as may be de wood such as by dipping or brushing or, in more severe sired, uniformity of retention within the wood is vari applications where long service life is required such as able, elevated impregnation temperatures for purposes with telephone poles, railroad ties, marine pilings, mine of efficiency can sometimes not be used, deposition of timbers and the like, pressure impregnation processes some of the materials causes embrittlement of the wood, are frequently used. and green lumber cannot be impregnated unless it is first Basically, pressure impregnation processes are classi dried. fied as either "full-cell' or "empty-cell' processes. The 10 essential difference between the two processes lies in SUMMARY OF THE INVENTION the fact that in the full-cell process, the liquid forced Accordingly, it is a principal object of this invention into the wood is retained by the wood after impregna to provide a method for the preservation of wood in tion. In contrast to this, in the empty-cell process, most which the wood is treated with an antimicrobial mate of the treatment fluid is expelled from the wood after 15 rial that is effective in preserving the wood and is com impregnation. The terms "full” and "empty" derive paratively nontoxic toward animal and plant life. from the fact that the cells of the wood are substantially Another object of this invention is to provide a filled with impregnants in the full-cell process, but tend method for the preservation of wood with a compara only to be coated with the impregnant in the empty-cell tively nontoxic but effective antimicrobial that will process. 20 become affixed or substantive to the wood. Another The full-cell process makes use of a vacuum/pressure object of this invention is to provide a method whereby impregnant cycle in which the wood is first placed wood can be treated with a preservative that is compar under vacuum and then, without admitting air, the atively nontoxic and can be applied utilizing standard treatment vessel is filled with the treatment liquid. After treatment techniques including dipping, brushing, and the wood is fully immersed in the liquid, the pressure is 25 full-cell and empty-cell pressure processes. increased to perhaps ten atmospheres or so and the Another object of this invention is to provide a liquid is forced into the wood. After the wood has been method for treating wood with a preservative in which treated to refusal, or until a predetermined gross absorp the preservative becomes affixed to the wood, the uni tion of the treatment liquid has been achieved, the pres formity of retention is improved, embrittlement of the sure is relieved and the treatment fluid is drained from 30 wood is avoided, elevated temperatures can be used the vessel. Usually a short vacuum cycle follows to during treatment, diffusion of the preservative into remove excess fluid from the surface of the wood. green lumber can be achieved, the danger of working The distinctive feature of the empty-cell process is with toxic chemicals can be avoided, and higher gross that it does not make use of an initial vacuum, but obsorptions and uniformity of retention can be im rather, as most commonly and preferably practiced, the 35 proved. wood is placed under pressure prior to the time that it is Briefly, these and other objects of this invention are contacted with the treatment liquid. The initial pressure achieved by contacting wood with a solution comprised is maintained during the time the treatment vessel is of an antimicrobial agent that is dissolved in a disubsti filled with the treatment liquid, and then the pressure is tuted aryl compound having an oleophilic and a hydro increased to a second higher pressure, forcing the treat philic substituent. In a preferred embodiment of this ment liquid into the wood against the air pressure ini invention, for example, the antimicrobial agent can be tially established within the wood. As a result, when the an organometallic complex such as a metal chelate of pressure is relieved, the air compressed within the wood 8-hydroxy-quinoline and the disubstituted aryl com expels much of the liquid that was forced into the wood. pound can be an alkylbenzene acid such as a commer This expelled liquid is referred to as the “kickback'. 45 cial grade of DDBSA. Commonly, as is the case with the full-cell treatment, a Use of the herein disclosed wood preservative solu vacuum is pulled in the treatment vessel after the im tions find considerable utility in the method of this in pregnation pressure has been relieved to increase the vention by improving significantly on some of the de recovery of the treatment liquid and shorten the period fects and encountered with the commonly available of time in which liquid will drip from the surface of the 50 wood treatment materials now in general use, such as wood. PCP, CCA salts and creosote. In somewhat more detail The empty-cell process is especially advantageous as than as briefly mentioned above, these improvements compared with the full-cell process when treating wood over the prior art include the following: with water-borne materials since the wood, after treat The compositions of this invention, when applied to ment, is ready for economic shipment, further treat 55 wood, have a high degree of fixation to the wood. Al ment, or immediate use since there is no large quantity though the preservative compositions as applied to or of water which must be removed as by kiln or air dry impregnated into the wood are solutions, many of the 1ng. compositions, including those based on a metal chelate Numerous disadvantages attach to the use of the most of oxine, become water-insoluble after application and commonly used wood preservation materials, not the drying and become highly fixed or substantive to the least among them being the more recently recognized wood the result is that they are resistant to removal dangers to the environment and to persons and animal even under severe water leaching procedures. life that come in contact with the treated wood. Gov It is known in the wood technology that when non ernmental tolerance for the PCP, CCA salts and creo polar solvent carriers, such as petroleum oils are used; sote treatment materials only exists due to the complete 65 these carriers penetrate the wood more quickly and absence of any alternative preservation material of provide higher retention and deeper penetration into proven effectiveness to take their place. The standard wood than preservatives utilizing polar carriers or sol treatment materials also suffer, to varying degrees and vents, such as water. The reason for the considerably 4,766,113 67 68 greater flow rate of nonpolar liquids is not understood, is seldom penetratd by the preservatives, especially in but one proposed explanation is that polar liquids, like the case of the hard woods, the upper temperature limi water, form strong hydrogen bonds which create fric tation imposed through the use of CCA salts does not tional drag between the wood and the liquid during permit good sterilization of the unpenetrated core. flow through the pores of the wood. Atwood moisture 5 Should decay fungi be present in the wood prior to contents below the fiber saturation point (about 30% treatment, these can continue to grow unchecked in the water on a dry wood base), flow rates of preservatives untreated portion of the wood. This may be of little in water carriers are reduced up to 50% as compared to consequence if the untreated center is small cross sec petroleum oils as reported in typical studies that have tion, since the major contributor to wood strength is in been published. Additional published studies of water 10 the outer treated portion of the cross section, but in the borne preservatives indicate that a number of these in case of species of wood that contain comparatively 0.5% concentrations in water exhibit average retention large, generally impenetrable section of heart wood, the reductions of up to 57% compared to the retention of loss of the untreated center to decay can bring about water alone when wood is treated via full-cell pressure rapid wood failure in service. The compositions of this procedures. It has also been indicated that the addition 5 invention, on the other hand, have no practical limita of a surface active wetting agent to water or water tions as to temperatures to which they can heated in the borne preservatives does not provide a to be expected treatment of wood, being limited only by the upper limit increase in flow rate through the wood. In point of fact, to which the wood itself can be heated without damage the flow rate is reduced when surfactants are used. It which is generally considered to be about 245 F. has been proposed that by reducing the surface tension 20 As noted above, the empty-cell process has many of the water, the rate at which the wood is swelled by advantages since the wood, after treatment, is not left the water becomes more rapid, with an attendant reduc saturated with the treatment liquid. Empty-cell pro tion in the size of the pores or passage through which cesses cannot economically be practiced with any de the water must flow. Unlike some of the compositions gree of efficiency with water-borne preservatives as of the prior art, those of this invention do not behave in 25 typified by the CCA salts. These salts are incompatible accordance with the above observations or theory of with the water-leachable materials in the wood and, as flow. The subject water-borne compositions penetrate a result, the contamination contained in the kickback wood more rapidly, not less rapidly, than do oil-borne (particularly solid soluble sugars) causes precipitation preservatives, and yield higher retention in the wood within working solution of sludge-like deposits that using any given method of application to the wood 30 render the working solution useless for further treat involving significant contacttimes between the wood ment. Not only does this represent a significant eco and the preservation solution. nomic loss, but further, due to the toxic nature of the The preservatives of this invention provide for CCA salts, a difficult disposal problem is presented. For greater uniformity of retention. Wood is a natural mate this reason, water-borne preservatives, such as CCA rial which is subjected to wide variations in physical 35 salts, are generally impregnated into the wood using properties, not only from species to species, but within full-cell processes, despite their disadvantages, rather different portions of a single piece of wood. For this than accept the problem of dealing with the kickback reason, piece-to-piece and within-a-piece retention of that results from empty-cell processes. However, in the preservatives is erratic and not always satisfactory. method of this invention, there is no incompatibility Published information indicates that in the case of creo 40 with the compositions of this invention and the materi sote end PCP, the retentions in a given pressure treating als leached from the water so that no unmanageable charge of wood may vary as much as from 1 to 4 and precipitates are collected in the kickback. Thus, in the sometimes even higher. Thus, to obtain a typical reten practice of this invention, empty-cell processes may tion, it is necessary to accept the fact that many pieces conveniently be used, gaining the inherent advantages of the wood will be grossly undertreated. Treating to a 45 of an empty-cell treatment with a water-borne high minimum retention specification is not economically -performance preservative not readily attainable in the feasible since the cost of greatly over-treating the bulk prior art. of the wood will be required. Thus, it is the industry Certain wood preservatives, and particularly water practice to treat only to an average which undoubtedly borne salts, tend to embrittle the wood to a degree that is an important cause of premature failures in service of, 50 depends on several factors including the level of salt for example, utility poles, railroad ties and the like. The retention in the wood. No such embrittlement is im compositions of this invention exhibit a much narrower parted to the wood in the practice of the instant inven range of retention than creosote and chlorophenol solu tion, and this may be of considerable significance, such tions. The improvement is substantial and provides as in the preservation of railroad ties, where continued obvious performance and economic advantages. Prema 55 impact of high-speed rolling stock may cause rail tie ture failure in service from undertreatment is reduced failure due to the brittle nature of the wood prior to the and the improvement allows significant reduction in time that the ties would fail from microbial decay. average retentions at a considerable cost savings. It is conventional to dry lumber prior to any pressure Certain of the prior art wood preservatives, such as impregnation treatment since green wood, having its CCA salts, are temperature sensitive in that they will cells filled with water, is essentially saturated and can precipitate rapidly from solution when heated and in not accept impregnating fluids. Water-soluble preserva contact with wood. For this reason, it is known to the tives can enter the wood by diffusion, but this is, at best, industry that CCA salts should not be used in wood unreliable, and at worst, of little value except at the preservation processes at temperatures above about surface of the wood. The requirements for a water-solu 120 C. This temperature limitation carries two draw 65 ble preservative that retains its solubility during a backs: (1) higher operating temperatures which gener lengthy diffusion process as is required with diffusion ally allow a shorter treating cycle cannot be used, and impregnation, limits the choice of preservatives to only (2) since the greater of a reasonable sized piece of wood a few permanently water-soluble toxicants such as 4,766,113 69 70 borax and zinc chloride which really cannot be consid The average retention of the former preservative was ered high performance preservatives. On the other 31 pcf as compared to 42 pcf for the latter. Taking into hand, the compositions of this invention combine the water-solubility and sap compatibility prerequisites of account the specific gravity differences, the composi successful diffusion preservative with high performance tion of Example 1 exhibited an average retention 22% and resistance to leaching since, after drying, the preser higher than the PCP solution. vative of this invention becomes water-insoluble and Example 3 affixed to the wood. The following composition was prepared and water There are some applications of wood preservatives in diluted for use: which a preservative is applied or reapplied after a 10 structure, such as building, has been erected. Since it is Oxine 4.1 parts by weight physically impossible to use pressure impregnation Copper hydrate 1.4 parts by weight methods in these instances, either brush or spray appli DDBSA 64.0 parts by weight cation must be relied upon. However, since the brush or Methanol 30.5 parts by weight spray application generally will penetrate only the sur 15 Also prepared were solutions of PCP in a petroleum face regions of the wood, it is generally desired to in oil. Both the foregoing composition and the PCP solu crease the concentration of the active ingredients in tion were used to pressure impregnate 3'x3'x18" dry, these treatment solutions to as high a level as possible so smooth-surfaced southern pine stakes to seven reten that their lack of penetration will, in part, be compen tions in the case of the composition of the invention and sated for by their strength. With present preservatives, 20 to four retentions for the PCP ingredient concentration there as severe limitations on the strength of the preser in solution was adjusted to yield the desired retention vative treatment solution that can be used including, on the basis of an average of 42 pcf total solution pickup low preservative activity per unit of weight, high mam in the case of the above composition and 31 pcf in the malian oral or skin toxicity, high skin and eye irritation, case of the PCP solutions. The retentions hereafter are strong odor and color, subsequent unpaintability, flam 25 reported in terms of total weight of the above composi mability, and high viscoities. In contrast, in the practice tion and PCP. of this invention, these limitations to the use of the pre servative in highly concentrated solutions are generally A full cell vacuum-pressure treatment procedure was eliminated, which allows the application of solutions used. The retort charge per retention was 100 stakes. with preservation efficacy or perhaps about ten times 30 Treatment operating schedule was the same for both that of the present products per unit of weight. Due to types of preservative solutions in all retentions. the low toxicity, the use of such highly concentrated Weighing each stake before and after pressure treat solutions does not present a danger to the health of the ment yielded these results: operator. Example 1 35 Low to High Retention Range (pcf) Difference (%) The composition below was prepared. (A) Example 3 Oxine 8.2 parts by weight Charge #1 0,29 to 0.37 28 Copper hydrate 2.8 parts by weight Charge #2 0.32 to 0.42 3. DDBSA 59.0 parts by weight Charge #3 0.52 to 0.72 38 Propylene glycol methyl ether 30.0 parts by weight Charge #4 0.68 to 0.97 43 Southern pine sapwood stakes were impregnated Charge #5 1.00 to 1.28 28 Charge #6 1.05 to 1.43 36 with the composition diluted 1:49 in water to a total Charge #7 1.44 to 1.78 24 solution retention of 42 pcf(lbs per cubic foot of wood). 33 Average After air-drying for 30 days, the treated 3'x3'x18' 45 (B) PCP long stakes were cut into 1" lengths and boiled in water Charge #1 0.16 to 0.25 56 for 4 hours. In this extremely severe leaching test, loss Charge #2 0.27 to 0.49 81 of preservative from the wood averaged 12% of that Charge #3 0.32 to 0.67 109 originally in the wood as measured by retained Cu-8-Q Charge #4 0.43 to 0.92 14 content. 50 90 Average Example 2 The composition of the preceding Example was di The foregoing retention ranges represent the spread luted in water 1:19 (5% concentration). Smooth-sur from the lowest to the highest retention obtained in the faced southern pine stakes ("X"X18' long) were 55 treatment of each 100 stake charge. immersed in the solution for 3 minutes, as were stakes in The improvement in retention uniformity of the com a 5% PCP solution in mineral spirits. Gross solution positions of this Example compared to PCP is large, retentions in the wood averaged 1.7 pcf for the Example both as to individual charges and overall averages. Of 1 composition (1.0 specific gravity) and 1.2 pcf for the further note to the relatively uniform spread in the PCP solution (0.82 specific gravity). Giving effect for high-low differences from charge-to-charge in the case the difference in specific gravities of the two solutions, of this Example whereas in the case of PCP, the reten the weight retention of Example 1 solution was 16% tion range is progressively larger as the average reten higher than the PCP solution. tion increases. This is of significance since the composi Under indentical conditions of vacuum-pressure im tion of this Example is normally used in the 0.2 to 0.6 pregnation, a 5% solution of PCP in a petroleum sol 65 pcf range whereas the normal use retention of PCP is vent (specific gravity of 0.9) and a 4% solution (1.0 0.4 to 0.6 pcf. The average high-low spread, therefore, specific gravity) of Example 1 composition were em in the most-used retention range is 32% versus over ployed to treat dry southern pine stakes (3'x3' x 18"). 100% for PCP, 4,766,113 71 72 been an age-old problem. Some authorities have stated Example 4 that more people in the Middle Ages died of ergot poi The composition of Example 1 was diluted 1:9 and soning, a toxic fungal metabolite of rye cereal grain, 1:99 in water and heated to 210 F. for 24 hours in a than of the plague. Even today it has been suggested by closed reflux system. No appearance difference was some authorities that food poisoning may cause some noted before and after the heating period, either at 210 90% of all human illness. F. and after cooling the solutions to room temperature. Since foodstuffs are intended to be ingested by hu Freshly prepared 1:9 and 1:99 use dilutions were then mans and animals, it goes without saying that the safety maintained at 210 F. 24-hours with the solutions sepa of their use and low toxicity are essential consider rately containing green and dry southern pine wood (10 10 ations. This fact has severely limited the availability of gms of wood per 100 gms of test solution). No solution known antimicrobials for food use. The antimicrobials appearance changes were noted. that currently find favor and are sanctioned for use Southern pine sap was obtained by compressing generally are of a low order of efficacy which fre fresh-sawn green pine at 2000 psi and collecting the quently results in a high cost-to-performance ratio, par discharged sap. A 1:49 use dilution of Example 1 com 15 position was prepared and admixed with pine sap in ticularly when used on raw foodstuffs for preservation. three weight ratios: 1 to 2, 1 to 1, and 2 to 1. After Mycotoxin formation in raw plant foods is a cause of 4-hours heating at 210 F., no appearance charge was growing concern. Those produced by certain Aspergil noted. After 3-month storage at room temperature, no lis, Penicillium, Alternaria, Trichoderma, Helminthos appearance charge was noted. 20 porium and other fungi are among the most toxic sub All solutions, before and after the test procedures stances known to man and are potent carcinogens, te were transparent, true solutions. All solutions after the ratogens and tremorgens. Proper handling and storage test procedures exhibited no significant change in active conditions drastically reduce, if not eliminate, these ingredient content. fungi, and hence the production of dangerous toxins. But even in the industrial nations, it is virtually impossi Example 5 25 ble to effect adequate control over hundreds of millions Fresh cut green southern pine sapwood stakes of tons of agricultural products which are grown, han (1" x 1"x72" long) were dip immersed into an 8% dled and stored by millions of persons. The result is water solution of the Composition Example 1, in a verti varying degrees of fungal growth on a significant por cal position, allowing the top 24' of the stake to remain 30 tion of the total. Subsequent processing generally de above the level of the solution. After one-week, the stroys the fungi but not the mycotoxins. stakes were removed and cut into 6' lengths and the Incidence of adverse effects on humans of mycotox cross-section of each piece qualitatively checked for ins in food is difficult to assess in the high technology depth and uniformity of preservative penetration. All nations, but is readily apparent in a number of the unde pieces originally immersed in the solutionn exhibited 35 veloped tropical areas where very favorable mold complete and essentially uniform penetration as mea growth conditions are coupled with few safeguards. sured by color change from natural wood to black, from Incidence of animal, particularly livestock, disease the placement on the wood of a 1% dithio oxamide (a from both fungi and mycotoxins usually is higher than copper indicator) in isopropanol. for humans. Some livestock and poultry diseases, In addition, preservative penetration completely caused by consumption of moldy feed, cause serious through the cross section of the stakes occurred up to economic losses. Aflatoxin M, an extremely toxic myco about 10" above the point of immersion. toxin, has been found in the milk of cows that have eaten moldy grain. SECTION 9 OBJECTS OF THE INVENTION According to another embodiment of our invention 45 we have discovered a method for the preservation of Accordingly, it is an object of this invention to pro foodstuffs and the prevention of mammalian diseases vide methods for the treatment of raw and prepared caused by contaminated food which involves using the foodstuffs that are effective to prevent food deteriora composition of Section 1 (hereinafter referred to as "the tion and diseases in mammals through the use of antimi compositions of this invention'). 50 crobial compositions that are generally nontoxic to More particularly, this invention relates to the preser animal and plant life. vation of foodstuffs and to methods for the treatment of Another object of this invention is to provide for the raw and prepared foodstuffs as well as food handling use of antimicrobial compositions that are compara equipment and packaging materials to prevent spoilage tively nontoxic to animal and plant life, but which are and mammalian diseases caused by pathogens. 55 highly efficacious in destroying microorganisms that cause food spoilage and disease-causing organisms. DESCRIPTION OF THE PRIOR ART Briefly, these and other objects of this invention are For many years antimicrobial compositions have achieved by treating foodstuffs and handling equipment been used in the treatment of foodstuffs to prevent their and packaging materials that come in contact with deterioration and to prevent diseases in humans and foodstuffs with a composition according to Section 1. animals that may be caused by food contamination. These antimicrobials have been used not only in treat DESCRIPTION OF THE INVENTION ing raw and finished food products, but also to prevent The compositions of this invention have unique util contamination that may be occasioned by the use of ity for foodstuff preservation by reason of safety, anti unclean handling and transportation facilities and pack 65 fungal and antibacterial efficacy that may be orders of aging materials. magnitude superior to current materials, and has virtu The problem of the preservation of food and the ally no color, odor or taste in use concentrations. Addi danger to mammals caused by contaminated food has tional features include residual persistence, low volatil 4,766,113 73 74 ity, high heat and sunlight stability and water insolubil ity after application, and excellent economy of use. Example 2 Among the suitable preservative applications for The following composition was tested for minimum compositions of the invention, the following may be fungicidal concentration and compared to two well mentioned: known antifungal chemicals-pentachlorophenol and (1) Fresh fruit, vegetables and nuts; 2,3,5 trichloro-4-propyl-sulfonyl pyridine-and (2) Dried fruits and vegetables during the dehydra DDBSA. tion process; Oxine 8.2 parts by weight (3) Cereal grains for human, animal and poultry con Copper hydrate 2.8 parts by weight sumption; 10 DDBSA 59.0 parts by weight (4) Animal silage; Propylene glycol methyl ether 30.0 parts by weight (5) Prepared foodstuffs; All of the test composition concentrations to be (6) Meat, fish and dairy products; tested were incorporated in the fungal growth media (7) Fruit and vegetable juices and fruit condiments; (agar) in accordance with standard microbiological and 5 practices. Agar plugs containing the test fungicides then (8) Handling and processing equipment, storage and were inoculated with a sporulating culture and inocu transportation facilities, and packaging materials lated at the temperatures and times specified by The for foodstuffs. American Type Culture Collection (ATCC) recom The most used preservatives typically are sanctioned 20 mendations. The plugs were then scored for absence or in the 100 to 1,000 ppm concentration range in food presence of organism growth. The results are shown in stuffs. These preservatives have acute oral LD50 toxicit the following table. Minimum fungicidal concentrations ies in the 100 to 5,000 mg/kg range. By contrast, the were determined against a broad spectrum of fungi that concentrates of this invention, exhibiting an LD50 of are detrimental to man, foodstuffs and materials and 1,500, are effective generally for preservation in the 25 which can result in metabolite formations (mycotoxins) range of 1 to 25 ppm against both fungal and bacterial of extreme toxicity to man and animals. attack. When the concentrates are diluted for effica In the table below, Composition A is that of this cious use, they may display LD50 toxicities above about Example 2 and the active ingredient is Cu-8-Q; Compo 20,000 mg/kg. Therefore, the compositions of the in sition B is DDBSA; Composition C is pentachlorophe vention have considerably lower toxicity in adequate 30 nol; and Composition D is 2,3,5-trichloro-4-propylsul use concentrations than present food preservatives. fonyl pyridine. Where no concentration is listed, no The following examples demonstrate the utility of data is available. The stated value of '1' means 1 or this invention for a variety of food preservation applica tions: fewer ppm. 35 EXAMPLES Minimum Fungicidal Concentration Example 1 - (ppm of active ingredient) The following composition was prepared and tested A. B C. D Aspergillis niger l 10,000 1-3 - for control of fungal growth on an outside, exposed-to (ATCC 9642) the-weather concrete slab which was covered com Aspergilis terreus 100 wom 36 pletely at the start of the test with black fungal growth (ATCC 10609) of unknown species (amounts are parts by weight): Aspergillis flavus 1 1,000 22-54 --- Oxine 4.4 (ATCC 11655) Copper hydrate 1.5 Alternaria alternata -- - - 45 (ATCC 13963) DDBSA 64.0 Aureobasidium pullulans 1 100 - --- Methanol 30.1 (ATCC 16624) After 1:150 use dilution in water, the composition Lenzites trabea 1 100 1-3 - (ATCC 11539) was applied to the concrete slab by spraying to run-off. Polyporus tulipiferae 100 1-3 - Within six months the black fungal growth had com 50 (ATCC 11245) pletely disappeared and no regrowth was noted during Penicillium breyl 1. 100 - - a two-year observation period thereafter. The six compactum (ATCC 16024) month period to initially remove the fungi is believed to Rhizopus stolonifer 1. 100 1-3 - (ATCC 24794) be that required for the dead organisms to dry, break up Trichodernia viride 10 100 - - and become unadhered to the concrete, and finally, to 55 (ATCC 8678) be washed away by rain. Trichoderma sp. 1 00 - In summary, the compositions of this invention pro (ATCC 12668) vide both a spectrum and a magnitude of performance Candida albicans 1 1,000 3 against plant and mammalian pathogens seldom seen in (ATCC 10259) any antimicrobial and heretofore unknown in one safe 60 enough for foodstuff use. These results illustrate the high efficacy of the com position of this invention and confirm the fact that an Examples 2-9 antifungal composition prepared from a Cu-8- Examples 2 through 9 that follow illustrate the broad Q/DDBSA solution is much superior to DDBSA alone. spectrum of the compositions of this invention for con 65 The results also indicate the favorable relative efficacy trolling fungi that cause foodstuff spoilage, that are of the test composition compared to the two commer pathogenic to mammals, and that produce dangerous cially available fungicides of recognized high perfor mycotoxins. aCe. 4,766,113 75 76 T. mentagrophytes-at 1:750 use dilution in a water Example 3 carrier, no growth after 10 minutes' exposure. The purpose of this test was to determine antifungal Simular but somewhat lower efficacy results were efficacy of DDBSA/Cu-8-Q solutions and to compare obtained by substituting zinc-8-Q or aluminum-8-Q in their efficacy to that of a world standard, sodium penta the composition of this example, produced by reacting chlorophenate, and a mixture of two well-known agri zinc oxide and aluminum hydroxide respectively with cultural fungicides, Topsin M (a thiophenate) and OX10. Nabam (a thiocarbomate). The test method is desig nated as a proposal for the Finnish NWPC Standard Example 5 No. 1.4.1.3/1974. The test substrate was fresh cut, green O The compositions of this invention exhibit efficacy pine wood. The test fungi were: against a broad spectrum fungal plant pathogens, as Blue stain fungi-mixture of Ceratocystis pilifera Z11 illustrated by various use dilutions in a water carrier of Sclerophoma entoxylina Z17 Pullularia pullulans U2 the following composition: Mold fungi-mixture of Paecilomyces varioti X15 Copper hydrate 1.70 parts by weight Cladosporium sphaerospermum R7 Aspergillis am 15 8-hydroxy quinoline 4.44 parts by weight stelodani X19 Isopropanol 35.00 parts by weight In the table below, Composition A is formulation DDBSA 58.86 parts by weight comprised of 64 wt.% DDBSA, 5 wt.% Cu-8-Q and 31 A. Valencia Oranges wt.% methanol diluted 1:200 with a water carrier. Tested on harvested fruit against Phomopsis stem Composition B is the same formulation diluted 1:100 20 end rot and Diplodis rot, at a 1:100 use dilution, 2 with a water carrier. minute dip application. After 3 weeks at 70 F., the Composition C is a 1.5% concentration of sodium following percentages of decay were noted: pentachlorophenate in water. Control (untreated) oranges-9.5% decay Composition D is a 0.4% concentration in water of a Treated oranges-5.3% decay 47:53 weight ratio of Topsin M:Nabam. 25 B. Sugar Cane E refers to untreated control pine boards. An agar seeding test against Ceratocystis paradoxa The rating index is: (pineapple disease) at a 1:10,000 (100 ppm) use dilution 0=no visible growth yielded a 3.0 mm. inhibition zone. l=traces of growth C. Peach Trees 2=slight growth 30 Tested against Taphrina deformans (causes leaf curl 3= moderate growth disease). Four test trees were sprayed twice, two weeks 4=covered with fungi apart, with a 1:400 use dilution. Three months later, 100 leaves on each test tree were rated for leaf curl: Control (untreated) leaves-100% leaf curl Test Composition 35 Treated leaves-13.5% leaf curl A. B C D E D. Cotton Blue stain fungi growth 0.8 0.3 0.5 2.5 3.9 Effectiveness against 11 fungi and 1 bacterium (Xan Mold fungi growth 0.5 0 0.3 1.3 3.6 thomonas malvacearum) that are associated with disease of cottonseed, seedlings and other plants was evaluated The demonstrated efficacy of Compositions A and B 40 in vitro, using the following compositions: against the six listed fungal organisms has utility not only on the tested substrate-wood-but also for pro Composition #1 Copper hydrate 1.70 parts by weight tection of a variety of other materials that are attacked 8-hydroxy quinoline 4.44 parts by weight by one or more of the fungi, including paint, concrete, Methanol 4.00 parts by weight brick, textiles and leather. 45 Isopropanol 30.86 parts by weight DDBSA 59.00 parts by weight Example 4 Composition #2 Copper hydrate 2.80 parts by weight 8-hydroxy quinoline 8.20 parts by weight Using the standard AOAC fungicidal test method Methanol 4.00 parts by weight (12th Edition, 1975), the composition below was evalu Isopropanol 26.00 parts by weight ated against two widespread fungi. 50 ODBSA 59.00 parts by weight Oxine 2.08 parts by weight Copper hydrate 0.70 parts by weight Both compositions were prepared in accordance with DDBSA 40.00 parts by weight procedures stated in previous examples. Isopropanol 32.00 parts by weight The following results were obtained, expressed in Demineralized water 25.22 parts by weight 55 parts per million (ppm) of total test composition in The two fungi were Aspergillis niger, a ubiquitous water carrier and the relative growth inhibition pro black fungus which flourishes on a broad range of sub vided at each test strength on each tested organism. In strates, and Trichophyton mentagrophytes, a cause of the tables below: "athlete's foot.' 0=no apparent inhibition A. niger-at 1:200 use dilution in a water carrier, no 60 1 s some inhibition growth after 10 minutes' exposure. 2=considerable inhibition (little growth) 3 = total inhibition (no growth)

Composition #1 Composition #2 Concentrations (ppm) Concentrations (ppm) Test Organisms O 5 25 100 500 1000 0 1 5 25 100 500 000 Pythium ultinum (41B) 0 0 0 1 2 3 3 0 0 2 2 2 2 4,766,113 -continued Composition #1 Composition #2 Concentrations (ppm Concentrations (ppm Test Organisms O 5 25 100 500 1000 0 1 5. 25 100 500 1000 Rhizoctonia solani (1D) 0 0 0 1 2 3 3 0 0 1 2 2 2 2 Fusarium (4A) 0 0 O 3 3 3 3 0 0 1 l 2 2 Fusarium (4D) 0 1 1 2 2 2 3 0 0 1 1 2 2 2 Fusarium roseum (4C) 0 0 0 1 2 2 3 0 0 0 2 2 2 2 Colletotrichum gossypii (35A) 0 0 0 1 2 2 2 0 0 0 2 2 2 2 Xanthomonas malvacearum (2A) 0 0 0 0 2 2 3 0 0 0 1 1 2 2 Aspergillis sp. l 1 Helminthosporium oryzae 2 2 Mucor mucedo 1 1 Penicillium sp. Rhizopus sp. 1. 1

Example 6 The composition of Example 4 was screened for fun- -continued gal pathogen response as a foliar spray on beans and Botrytis Alternaria rice. The ratings scale is from 0 (no pathogen control) to 20 Inhibition zone (mm) 13 14 16 0 0 4 10 (complete pathogen control). The concentration of active ingredient (in a water carrier) of all compositions tested is 33 parts per million (ppm). The active ingredi The DDBSA/Cu-8-Q solution of this invention ex ent in the composition of Example 4 is expressed in hibits an improvement in efficacy against the tested terms of Cu-8-Q and the chemical as listed below for 25 organisms by a facor of 4x in the case of Botrytis to 10x four comparative products. The comparative products in the case of Alternaria. tested were Karathane (2,4-dinitro-6-(2-octyl phenyl crotonate), Vitavax (5,6-dihydro-2-methyl-1,4-oxathin Example 8 3-carboxanilide), Daconil (tetrachloroisophthalonitrile) The DDBSA/Cu-8-Q solution of the preceding ex and Maneb (manganese ethylenebisdithiocarbamate). 30 ample (31), diluted 1:400 in a water carrier, was applied The plants and diseases tested were bean mildew by spray nine times, at two-week intervals, to peach and (Erysiphe polygoni), bean rust (Uromyces phaseoli typica) nectarine culivars during the growing season. The re and rice spot (Helminthosporium orazae and Cerocospora sults against brown rot (Monolinia fructocola), com orazae). pared to nontreated trees, is presented below. 35 Bean Mildew Bean Rust Rice Spot % Fruit Affected Example 28 composition 8 10 10 Peach Nectarine Karathane 10 - At harvest (treated) 3 Vitavax - 9 - At harvest (untreated) 15 43 Daconil --- 10 40 Five days later (treated) 3 6 Maneb - w 8 Five days later (untreated) 60 67

Example 7 Example 9 45 The following composition was evaluated (diluted The DDBSA/Cu-8-Q solution of Example 8 was with water for use) in vitro for inhibition against two tested in vitro against a major turf pathogen, Helmin fungal pathogens, Botrytis sp. and Alternaria sp., causal thosporium vagans, via a standard agar plate culture agents of a variety of plant diseases. technique, with these results: Oxine 2.08 parts by weight Copper hydrate 0.70 parts by weight 50 DDBSA 40.00 parts by weight Fungus Colony Diameter Triton X-100 20.00 parts by weight 1:6700 use dilution 1 mm Isopropanol 22.00 parts by weight in water carrier 1:3350 use dilution O Water (demineralized) 15.22 parts by weight in water carrier The zone of inhibition agar plate test also was used to Control 21 mm test Cunilate 2174 (diluted in mineral spirits for use) for comparison. The composition concentrations in the The results demonstrate very high efficacy in con table of results below are expressed in parts per million trolling this important pathogen. Complete control of (ppm) of Cu-8-Q. The larger the inhibition zone, the 60 H. vagans was achieved in this assay between 3.7 and 7.5 greater is the efficacy of the composition. ppm of Cu-8-Q. As illustrated in the examples to follow, the composi Botrytis Alternaria tions of this invention have high efficacy against abroad DDBSA/Cu-8-Q spectrum of bacteria that are pathogenic to mammals Concentration (ppm) 21 50 125 2. 50 125 65 and plant life, that contribute to reduced water quality, Inhibition zone (mm) 3 5 22 O 16 21 that cause deterioration of foodstuffs, that degrade a Cunilate 2174 broad range of manufactured and natural materials and Concentration (ppm) 83 200 500 83 200 500 products, and which generate toxic metabolites (bac 4,766,113 79 80 teria-toxins) that are among the most poisonous sub stances known to man. Example 10 Of particular interest is high efficacy against Gram The DDBSA/Cu-8-Q solution of Example 2 was negative as well as Gram-positive microorganisms. Few tested, along with a number of well known antimicro antibacterial materials now available are effective bial agents, against a broad spectrum screen of economi against the Gram-negatives and still fewer provide eco cally important Gram-positive and Gram-negative bac nomical control of them. A number of available antibac teria. All antimicrobial agents were incorporated in the terials toxic to Gram-negative organisms have practical agar bacterial growth media according to standard mi limitations which severely restrict use, including high crobiological practices. The bacterial species were mammalian toxicity, phytotoxicity, corrosiveness to 10 grown in nutrient broth; 24-hour cultures, the inocu skin and a variety of materials, strong odor, strong lum, then were streaked onto the nutrient agar plates color, high volatility, low or erratic shelf stability, low containing the test antimicrobials. After a 24-hour incu or nonexistent residual activity, and prohibition of use bation at the appropriate temperature, the plates were at elevated temperatures. rated for presence or absence of bacterial growth. The basic significance in the need for Gram-negative Minimum bactericidal concentrations for each of the control lies in the fact that this bacterial category in tested agents are stated in the following tabulation of cludes a number of widespread, virulent pathogens results in parts per million (ppm) of active ingredient as which are difficult to impossible to control with pres defined in the description of each agent.

Antimicrobial Agent (ppm of active ingredient) Bacteria (ATCC No.) A. B C. D. E F G Gram-positive: Bacillus cereus l 100 - 7 8 5-10 o Bacillus lichenforms (27326) l 100 - 7 8 2-5 al Bacillus megaterium (27327) 100 - 7 8 m Bacillus subtilis (37328) 1. 100 - 750 8 M 3 Micrococcus flavus (10240) l 100 - 7 8 - m Mycobacteriumphlei (15610) 1 10 - 7 8 - 3 Staphylococcus aureus (6538) l 100 2083 7 8 1-3 3

Alcaligenes faecalis (337) 10 1000 -- 750 80 MM Alcaligenes marshali (21030) 104 100 - 7 8 - Esherichia coli (11229) 104 10,000 -- 750 .80 250-500 165 Flavobacterium arboresceus (4358) 10 10 4166 7 8 - m Klebsiella pneumoniae (4356) 10 10,000 -- 750 8 m Proteus vulgaris 10 1000 - 7SO 800 m Pseudomonas aeruginosa (15442) 104 1000 4166 750 800 1000-2500 165 Salmonella cholerasuis (10708) 104 1000 -- 750 80 250-500 16S Salmonella typhi (6539) 104 1000 2083 750 80 m 65 'stated value of "" means or less A = the didBSA/Cu-8-Q solution of Example 2 with the active ingredient expressed in terms of Cu-8-Q. B = DDBSA. C = phenol D = Betadine, an iodine/polyvinylpyrrolidone complex containing 0.75% iodine. The active ingredient is iodine. E = Alkyl dimethyl ammonium chlorides (61% C2, 23% C, 11% C16 and 3%. Co). F = sodium pentachlorophenate G = 2,3,5-trichloro-4-propylsulfonyl pyridine. ently available antibiotics, notably Pseudomonas sp. typified by Pseudomonas aeruginosa PRD-10, the stan These data demonstrate the high efficacy of the com dard strain in the United States for evaluation of anti position of Example 2. On the basis of the average of the bacterials for mandatory Gram-negative control appli efficacies against all the test organisms, Composition A cations. 50 is 45 times superior to Composition B; 88 times better The compositions herein disclosed eliminate or sub than Composition C; 10.7 times better than Composition stantially reduce these use limitations inherent in many D; and 3.5 times superior to Composition E. other germicides. The compositions are quite unique in On the basis of the average of the efficacies against having strong Gram-positive and Gram-negative activ the three test bacteria (Staphylococcus aureus, Salmo ity combined with broad versatility of formulation and 55 nella cholerasuis and Pseudomonas aeruginosa PRD-10) use plus a high degree of safety (low toxicity and zero to required by the Environmental Protection Agency of a low skin and eye irritation). Add to this the high effi "hospital grade" disinfectant, Composition A is 10 times cacy, broad spectrum antifungal activity of the compo better than Composition B; 7.2 times better than Com sitions of this invention and the resulting range of toxic position D; and 1.6 times better than Composition E. ity to target organisms and safety to man, the most sensitive of hosts, is unique indeed. Example 11 The balance of toxicity provided by this invention to The composition set forth below was prepared by Gram-positive, Gram-negative and fungal microorgan previously described procedures: isms has special value in the broad consumer field of Oxine 2.08 parts by weight skin deodorancy. Present antibacterials suffer from the 65 Copper hydrate 0.70 parts by weight fact that they are effective primarily against Gram-posi Isopropanol 32.00 parts by weight tives, allowing Gram-negative and fungi overgrowth, a DDBSA 40.00 parts by weight condition considered dangerous by many authorities. Water (demineralized) 25.22 parts by weight 4,766, 113 81 82 When evaluated as a bactericide by the AOAC Use lated below and refer to plants and tubers produced Dilution Method (12th Edition, 1975), 10 ring carriers from the tubbed test seed pieces. per organism, the following results were obtained ( subculture and B =resubculture): Ring Rot % % % Yield Contam- Plant Ring Rot Ring Rot cwt/ Use Dilution Negative Positive Antimicrobial inated Stored Plants Tubers aCe In Water Carrier A B A B None (control) Yes 98 23 8 493 None (control) No 95 O O 609 Staphylococcus aureus 1:1000 10 10 O O DDBSA/ Yes 98 O 2 631 Salmonella cholerasuis 1:1000 10 10 O O 10 Cu-8-Q (PRD-10) DDBSA/ No 100 O 0 602 Pseudomonas aeruginosa 1:400 10 10 0 O Cu-8-Q Aerobacter aerogenes 1:400 10 10 0 O 20% Clorox Yes 98 20 9 500 20% Clorox No 98 O 0 602 Roccal Yes 98 20 9 515 A ten-minute kill is required against the first three Roccal No 98 O O 638 pathogens for sale as a hospital grade disinfectant. Effi 15 cacy against the fourth organism, a major cause of slime in recirculated cooling water systems and pulp and The composition of this example demonstrates supe paper mills, demonstrates utility of the composition as a rior control of the ring rot bacterium. Other species of slimicide. the genus Corynebacterium are causal agents of disease 20 in man and a variety of plant life. Example 12 Example 13 This composition was prepared and tested at one use dilution, 1:50 in water carrier, against the causal agent The composition below was prepared and tested for of potato ring rot bacteria (Corynebacterium Sepedoni speed and range of antibacterial activity, in the absence cum): 25 and presence of organic matter (blood) for use in hospi Oxine 2.08 parts by weight tal disinfection, cold sterilization and antisepsis. Copper hydrate 0.70 parts by weight Oxine 4.1 parts by weight DDBSA 40.00 parts by weight Copper hydrate 1.4 parts by weight Triton X-100 2000 parts by weight DDBSA 65.0 parts by weight Isopropanol 22.00 parts by weight 30 Propylene glycol methyl ether 29.5 parts by weight Water (demineralized) 15.22 parts by weight Many antimicrobial agents are partially or totally Infected potato seed readily contaminate potato seed deactivated in the presence of organic matter, constitut cutters, sacks, bins, cellars, trucks and planting equip ing a severe limitation to effectiveness of such agents for ment with the highly infectious ring rot bacteria. The a number of uses such as wound antisepsis and medical result may be infected potato plants, tubers and reduced 35 instrumentation and surface disinfection where large yields. amounts of organic matter often are encountered and The test procedure consisted of dipping unpainted, sometimes are unavoidable. planed wood laths (6') into a slurry of infected ring rot The AOAC Use Dilution Confirmation Test (12th tuber tissue, allowing excess slurry to drain off (3-5 Edition, 1970) was modified as follows: minutes) and then spraying the contaminated lath with (a) The test temperature was 37 C. the test antibacterials. Three to five minutes later, (b) The ring carriers were soaked in sheep blood for healthy Norgold Russet potato seed pieces were rubbed two hours, air-dried for one hour, then contami vigorously against both sides of the contaminated and nated with the test pathogen. antibacterial-treated laths. The process was repeated (c) The contaminated rings were contacted with the using laths not contaminated with C. Sepedonicum but 45 test antibacterial agent for 30-second, one-minute treated with the test antibacterial agent. The rubbed and three-minute periods. seed pieces were stored in bags and later planted at the The results are set forth in the following table, in appropriate time. which: In addition to the composition of the invention, un 0=no growth in 10 of 10 tubes tested treated controls, 20% Clorox (1.05% sodium hypochlo 50 1 = growth in 1 of 10 tubes tested rite in water), formaldehyde (37% formalin diluted 2=growth in 2 of 10 tubes tested 1:120 in water) and Roccal (benzalkonium chloride or 3=growth in 3 of 10 tubes tested etc.

Use Dilution in In Absence of Blood In Presence of Blood Test Pathogen Water Carrier 30 sec. 1 min. 3 min. 30 sec. 1 min. 3 min. Staphylococcus aureus Undiluted O O O O O 0. (ATCC 6538) 1:10 0. O O O 0 0. 1:100 0 O O 3 6 3 Salmonella typhi Undiluted O 0 O O O 0 (ATCC 6539) 1:10 O O O O 0 0 1:00 O O 0 O O 0 Pseudomonas aeruginosa Undiluted 0 O O O 0 0 (ATCC 15442) 1:10 0 O 0 10 10 0 1:00 O O 0 10 3 3 These results indicate that the test composition is Zephiran chloride) diluted with water to 800 ppm con capable of rapid antibacterial action in the presence of centration were tested. The results of the test are tabu substantial amounts of organic matter against the three 4,766,113 83 84 human pathogens generally considered as definitive for with Cunilate 2174, a commercially available concen antibacterial efficacy evaluation. trate containing 10% Cu-8-Q which is made soluble in Example 14 petroleum hydrocarbon solvents via use of nickel ace tate and 2-ethyl hexoic acid. The previously described The following DDBSA/Cu-8-Q solution was pre Nylate 10 also was tested. pared in accordance with previously stated techniques. Oxine 4.1 parts by weight (In this instance, Cu-8-Q was formed in situ from copper Copper hydrate 1.4 parts by weight hydrate and oxine, also known as 8-hydroxyquinoline.) DDBSA 64.0 parts by weight Copper hydrate 1.70 Propylene glycol methyl ether 30.5 parts by weight Oxine 4.44 10 DDBSA 64.81 This composition was use-diluted 1:24 with a water Methanol. 15.05 carrier; the Cunilate 2174 was diluted 1:19 and 1:9 with Isopropanol 14.00 mineral spirits for use; the Nylate 10 was diluted 1:19 This composition was diluted with a water carrier, as and 1:9 with water. The cotton samples were dipped to tabularized below, and tested in comparison with a 5 refusal in the test compositions, air-dried and buried at sodium tetrachlorophenate (23%) liquid concentrate, 75' F. for 29 days in sheep manure moistened with also diluted in a water carrier, against organisms on water. Microorganism attack on the cotton cloth in this three species of green lumber-Douglas fir, Amabilis fir test medium is both rapid and severe as can be noted and Ponderosa pine. The organisms were: from the essentially total destruction of the untreated Cephaloascus fragans a brown mold that infects cer 20 control cloth sample. The results of this test are tabu tain wood species lated as follows: Trichoderma virgatum a common cold Mixed spores a combination of two molds (Penicil lium sp. and Aspergilis niger) and a fungus (Cerato Weight of Cu-8-Q Estimated cystis pilifera that causes blue stain in wood. Composition (dilution) in Cloth Strength Loss" 25 Untreated control O 100% The freshly cut wood samples were dip treated (15 Cunilate 2174 (1:19) 0.41 gram 50% second immersion) with the test fungicides and then Cunilate 2174 (1:9) 0.84 gram 25% innoculated with spore suspensions of the above Nylate 10 (1:19) 0.61 gram 50% described fungi. The test boards plus untreated control Nylate 10 (1:9) 1.21 gram 0% boards were then placed in a warm, humid chamber for 30 DDBSA/Cu-8-Q (1:24) 0.18 gram 0% four weeks. The results are set forth in the table below As measured by tear strength reduction: 100% = total loss of strength in which: 50% = moderately difficult to tear by hand 25% = difficult to tear by hand 0% = impossible to tear by hand Mixed Use C. fragans T. virgatum - spores 35 The superiority of the formulation of the invention Dilution A B C A B C A B C over other Cu-8-Q compositions is clearly evident. Tetra- 1:100 4 4 4 0 O 3 3 4 4 chlorophenol composition SECTION 10 DDBSA/Cu-8-Q 1:240 2 0 1 3 0 4 4 3 4 40 According to another embodiment of our invention solution Tetra- 1:50 2 0 4 0 0 1 3 2 4 we have discovered that the compositions of Section 1 chlorophenol (hereinafter referred to as "the compositions of this composition invention') can be used to reduce the population DDBSA/Cu-8-Q 1:120 0 0 1 3 0 1 2 2 2 growth of microorganisms. solution Tetra :25 2 2 3 0 0 1 0 0 3 45 DESCRIPTION OF THE PRIOR ART chlorophenol composition Numerous antimicrobial agents are known to the DDBSA/Cu-8-Q 1:60 0 0 0 1 0 O 0 0 0 solution prior art for use in treating water for sensitizing, steriliz Tetra 1:12.5 0 1 0 0 O 0 0 O 2 ing or reducing the growth of microorganisms. These chlorophenol 50 treatments find utility for such purposes in conditioning composition circulation cooling water, waste water from pulp and DDBSA/ 1:30 0 0 O 0 O 0 0 0 0 Cu-8-Q solution paper processing, effluence from sewage treatment Control 4 4 4 3 4 4 - 4 4 4 plants, water in swimming pools, drinking water, and (no treatment) many various discharges and wastes. One of the pri A = Douglas fir 55 mary difficulties encountered in utilizing the prior art B = Annabilis fir C = Ponderosa pine antimicrobial agents lies in the fact that many of them 0 = no growth are quite toxic to animals and plants, and even the less st 2 = medium growth toxic ones, when mixed in water in amounts necessary 3 = heavy growth in 4 weeks to obtain suitable performance, may irritate tissues such 4 = heavy growth in 2 weeks as the skin and eyes, and they may impart odors or tastes to the treated water. Further, many of the prior art antimicrobials have a narrow spectrum of antimicrobial Example 15 activity which frequently results in controlling certain To further illustrate the antifungal properties of com specific organisms at the expense of permitting over positions of the invention, the formulation below was 65 growth of other organisms by the removal of checks prepared and tested as a preservative against microbio and balances. They also suffer in that they are often logical deterioration of 10 oz. cotton duck cloth and readily washed away and do not have high substantivity compared with untreated cotton duck as a control and to provide ongoing antimicrobial protection for water 4,766,113 85 86 treating equipment such as cooling towers, conduits, including materials such as cellulose (e.g., wood and vessels, basins and the like. natural fabrics), concrete and bricks. This is of particu lar value in reducing microorganisms in water-treating OBJECTS OF THE INVENTION equipment since the adherence of the antimicrobial Accordingly, it is an object of this invention to pro agents to various surfaces will provide some degree of vide a method for treating water to eliminate, reduce or permanence in preventing the growth of microorgan otherwise control the microbe population in the water isms in water-treating equipment, pipelines carrying through the use of compositions that have low toxicity water, and holding and storage devices such as pools, to animals and plants and low irritability to the skin and basins, tanks and the like. Particularly noteworthy of eyes. 10 applications in which this invention finds ability is in the Another object of this invention is to provide a treatment of water such as recirculating cooling water, method for the treatment of water and other substrates pulp and paper processing, sewage effluent control, for purposes of sanitization and sterilization through the sterilization or sanitation of drinking water, swimming use of comparatively nontoxic, odor-free, nonirritating pool treatment, and the general treatment of effluence compositions. 15 and waste water to avoid environmental contamination Another object of this invention is to provide a and prevent objectionable odors from being generated method for inhibiting the growth of microorganisms in by bacterial and fungal processes. water by treating the water with compositions that are The following examples illustrate the value of this comparatively nontoxic, odor-free and nonirritating. invention in selected water treatment applications: Another object of this invention is to provide a 20 method for the sterilization of water and the inhibition EXAMPLES of microorganisms that, through biological action, pro Example 1 duce unwanted odors. These and other objects of this invention are For swimming pool use, antimicrobials serve two achieved by treating water, such as waste water, recir 25 purposes: (1) elimination or reduction of pathogenic culating water, process water, swimming pool water, flora to an acceptable level, and (2) elimination or re and drinking water, with antimicrobial agents, solubi duction of organisms that degrade the aesthetic appeal lized with a disubstituted aryl compound having hydro of the water, the container (pool walls and floor), and philic and oleophilic substituents. In one preferred em the surrounding area (diving boards, steps, safety rails bodiment, the antimicrobial agent is an organometallic 30 and mats, etc.). Algae, nonpathogenic bacteria and fun compound, such as a metal chelate of 8-hydroxy-quino gal growths, for example, will turn clear, sparkling line (oxine), and the disubstituted aryl compound is an water into tubrid, slimy water. alkylbenzene sulfonic acid such as, for example, a com The most common antimicrobial for pool use is free mercial grade of dodecyl benzene sulfonic acid available chlorine supplied by a variety of compounds, (DDBSA). 35 the most widely used of which is calcium hypochlorite. This source of chlorine presents a number of complica DESCRIPTION OF THE INVENTION tions in use, including need to maintain critically close The utilization of the antimicrobial compositions of water pH control and reaction with nitrogen-containing this invention are especially distinguished from prior art substances in the water to form objectionable reaction compositions due to their high efficacy against a broad 40 products such as chloramines which irritate the eyes of spectrum of microorganisms while at the same time bathers. Also, a chlorine stabilizer may be needed to being comparatively nontoxic and nonirritating to ani inhibit ultraviolet light degradation, and an algeal con mal and plant life. The antimicrobial compositions of trol agent. this invention have high efficacy against a broad spec The use of the compositions of this invention substan trum of microorganisms including both Gram-positive 45 tially simplifies pool treatment. This reduces the oppor and Gram-negative bacteria, the four major classes of tunity of improper maintenance and attendant health fungi (Ascomycetes, Basidiomycetes, Phycomycetes risks. Need for close pH control of the water is elimi and Fungi imperfecti), algae, protozoa and viruses nated, sunlight stability is satisfactory, and algeal as well (naked and envelope types). as bacterial control is provided. The compositions also An unusual feature of the compositions used in the 50 provide satisfactory efficacy against Alcaligenes faecalis method of this invention is that they not only have which, though not considered generally to be a true broad antimicrobial activity, but they exhibit generally pathogen, is undesirable and can interfere with interpre low mammalian toxicity and skin and eye irritation. tation of total count tests for sanitary quality of the When mixed in water for suitable performance in re Water. quired amounts, they generally may be considered to 55 Using standard microbiological antimicrobial testing have essentially zero levels of toxicity and skin and eye techniques, the following composition was evaluated irritation. against representative organisms involved in swimming Another advantage that accrues in the practice of this pool maintenance. Amounts are parts by weight. invention is that the antimicrobial compositions have Oxine 8.2 low odor and taste and impart a low color to treated 60 Copper hydrate 2.8 water. When mixed in water in concentrations for DDBSA 60.0 stronger than generally required for effective perfor Ethanol 10.0 mance, the water has general acceptability since the Propylene glycol methyl ether 19.0 compositions impart little or no taste, odor or color to the water. 65 Microbicidal Concen Use of the compositions of this invention is also of tration (ppm) (based utility since their active ingredients have specific affin on Cu-8-Q) ity (substantivity) toward a broad range of substrates Mycobacteriumphlei (ATCC 15610) 10 4,766,113 87 88 -continued Example 4 Microbicidal Concen tration (ppm) (based The following composition was prepared (amounts on Cu-8-Q) are parts by weight): Staphylococcus aureus (ATCC 6538) 10 Oxine 4.1 Alcaligenes faecalis (ATCC 337) 100 Copper hydrate 1.4 Esherichia coli (ATCC 11229) 1040 DDBSA 64.5 Klebsiella pneumoniae (ATCC 4356) 1040 Pseudomonas aeruginosa (ATCC 15442) 1040 Propylene glycol methyl ether 30.0 Salmonella typhi (ATCC 6539) 1040 A 1:400 use dilution in water was made up and 5% Candida albicans (ATCC 10259) 10 10 paper pulp admixed therewith. Analytical determina Aspergillis niger (ATCC 9642) 10 tion of the active ingredient retained in the solution was made after three periods of time: Example 2 15 Time after Intro Active Ingredient Reduction The following composition was prepared (amounts duction of Pulp in the Solution (based on Cu-8-Q) are parts by weight): 3 minutes 23% Oxine 4.1 1 hour 47% Copper hydrate 1.4 24 hours 73% DDBSA 64.5 Methanol 30.0 20 The composition was tested for microorganism con These results provide an indication of affinity of the trol in paper mill water. The water samples, with and cellulose pulp for the active antimicrobial ingredient. without addition of the test composition, were sub SECTION 11 jected to a microbial count in accordance with standard 25 According to another embodiment of our invention mill quality control procedures. we have discovered that the compositions of Section 1 (hereinafter referred to as "the compositions of this No. of Microbe invention') are useful for cleaning, sanitizing or disin Colonies/ml fecting substrates by contacting the substrate with such % Composition Before -hr. % 30 compositions. in Mill Water Aging Aging Reduction Control water O 102,000 175,000 m DESCRIPTION OF THE PRIOR ART Treated water 0.09 3,000 98.3 As a rule, disinfecting and sanitizing antimicrobials Treated water 0.19 0 100.0 are included within cleaning agents only for specialty 35 uses and the combination of antimicrobials with clean In addition to high antimicrobial efficacy, substantiv ing agents has comprised only a small portion of the ity of the composition to cellulose removes much of the over-all cleaner markets. A variety of factors have dic antimicrobial from the process water by affixation to tated against large-scale incorporation of antimicrobials the pulp. This serves two unique functions: (1) substan into cleaning compounds for general usage because of tially reduces the quantity of antimicrobial that other the increased costs, various problems that arise, and wise would be discharged with the paper mill effluent limitations that are imposed upon their general use. into the environment, and (2) provides, at no extra cost Representative of the various problems that may be over that required to treat the water, an antifungal agent encountered in the use of antimicrobials in cleaning contained within the final products of the paper mill. compounds are the taste and odor that they frequently 45 impart to the cleaning compound; the lack of flexibility Example 3 in using water and organic, polar and nonpolar carriers; To illustrate the substantivity of the compositions of the high danger of skin and eye irritation; high water this invention to cellulosic surfaces (pulp, paper and resistance; poor heat and light stability; poor substan wood) the composition of the preceding example was tive properties which render the antimicrobials tran diluted 1:500 with water and 5% green oak wood flour SO sient and readily washed away; loss of antimicrobial added and thoroughly mixed. Within five minutes, activity in the presence of organic matter and dirt; poor 20-30% of the active ingredients in the test composition shelf life and stability; and high toxicity toward animal became affixed to the wood particles as determined by and plant life when the antimicrobials are used in effica testing the active ingredient concentration in the solu cious amounts. tion. 55 In a similar test, a 1:5 water dilution of the composi OBJECTS OF THE INVENTION tion was prepared and a piece of smooth-surfaced, dry Accordingly, it is an object of this invention to pro southern pine wood dipped in it for five seconds, re vide methods for cleaning dirt from substrates with moved and washed under running water for five sec simultaneous sanitization, disinfection or sterilization. onds. A qualitative color test for copper (1% dithiooxa Another object of this invention is to provide a com mide in isopropanol) indicated the presence of copper position which may be used not only to clean substrates on the wood surface. from organic and inorganic dirt, but also to destroy or This unique substantivity has utility in protection of greatly reduce the population of microorganisms on the wood cooling towers and other wooden components of substrate. recirculated water cooling systems. These wood parts 65 Another object of this invention is to prepare a com are highly susceptible to fungal and bacterial degrada position for use in cleaning, sanitizing, disinfecting or tion and to surface coatings of slime which reduces sterilizing substrates, which compound has low odor functional efficiency. and taste, high residual resistance to microbial activity, 4,766,113 89 90 high resistance to loss of antimicrobial activity, high sanitizing, disinfecting or sterilizing compound has been resistance to loss of antimicrobial activity in the pres compromised to accommodate present practical reali ence of organic matter, substantivity to many substrates, ties. This is not true of the compositions used in the inherently high cleaning properties over a wide range practice of this invention since they generally avoid the of substrates, high water resistance, low toxicity toward 5 limitations of products available in the marketplace and animal and plant life, low to zero skin or eye irritation, make practical and safe an antimicrobial cleaner that long-term shelf life and stability, and high compatibility may be classified as a hospital-type disinfectant or disin with water and organic, polar and nonpolar carriers. fectant cleaner. Briefly, these and other objects of this invention are By way of explanation of terms, it is noted that sanita achieved by treating substrates with the compositions of 10 tion is used to suggest a reduction in pathogenic organ Section 1. isms of selective genera to safe levels. The sanitization classification generally denotes a considerably lower DESCRIPTION OF THE INVENTION level of antimicrobial action than products classified as The cleaning, sanitizing and disinfecting composi disinfectants which, in turn, usually denotes a lower tions of this invention are characterized in that they are 15 degree of antimicrobial activity than does sterilization. effective for use both as cleaning agents to remove In most applications, the composition of this invention organic and inorganic dirt and at antimicrobials to kill makes it practical to obtain disinfection instead of saniti or greatly reduce the population of microorganisms. zation and, in some cases, even sterilization instead of The antimicrobial activity of these compounds has a sanitization or disinfection. very high per unit efficiency against a large group of 20 Cleaning agents having antimicrobial properties are microorganisms that are pathogenic to man and ani generally recognized as providing superior odor con mals, that cause unpleasant odors, and that may cause trol than do cleaners alone. The use of the compositions the deterioration of a wide range of materials. The com of this invention allows a broad upgrading in cleaning positions of this invention are further unique in that they sanitization since a broad general improvement is have a high order of safety relative both to mammals 25 achieved in odor control such as may be caused by and plant life and to various substrates that are sensitive fungal or bacterial decomposition. While control over to damage from contact with acidic, alkaline or oxidiz bacterial odor is greatly improved by use of the compo ing chemicals. sitions of this invention, improvement in the control of The compositions of this invention also overcome the odors caused by fungi is even more significant due to inherent limitations of many of the cleaning, sanitizing 30 the lack of presently available antifungals that are safe, and disinfection compounds of the prior art in that they cheap, practical enough from a safety standpoint for have low color, taste and odor; have a high residual broad usage, and still efficacious against a broad spec resistance of antimicrobial activity; are usable in both trum of fungi. In contrast to the odor-controlling clean water and organic, polar and nonpolar carriers; have ing agents now available, the use of the compositions of low to zero skin and eye irritation characteristics; have 35 this invention are highly effective at practical yet non high water resistance; have high heat and light stability; toxic levels in controlling the odors that arise, for exam have inherently high surfactant, detergent and cleaning ple, in homes, farm buildings, commercial and public properties that may be modified with a large number of buildings, hospitals and institutions, animal quarters, commercially available detergent, surfactant and wet zoos, foodstuff processing facilities, transportation ting agents; are substantive agents to many substrates; 40 retain significant antimicrobial activity in the presence equipment, fishing vessels, industrial plants and ware of organic matter and dirt; have long-term shelflife and houses, subway facilities, mortuaries, restaurants, sew stability; and last, but by no means least, display rela age, hospital and food-processing organic waste, recir tively little toxicity toward animal and plant life. culation water, air systems, and on through a host of The economy and safety of using the compositions of 45 other environments in which odor control may be de this invention, plus the range of their desirable proper sired. ties, make it practical, unlike the compositions of the It should be noted that in the application of the com prior art, to incorporate antimicrobials in general clean positions of this invention to various substrates, the ing compounds on a widespread basis. This upgrades primary nature of the deodorant action is in the preven and increases the utility of general cleaning composi 50 tion and blockage of further odor generation by the tions as, through their use, not only will dirt be re destruction of microbial action and not in the destruc moved, but control will be gained over the population tion of existing odors. and growth of microorganisms that cause disease, odors To illustrate methods for practicing this invention and destruction of organic materials. and to demonstrate the use of the compositions of this The compositions of this invention are also unique in 55 invention, the following examples are given: their high efficacy to cost ratio. EXAMPLES It may be noted that three classes of disinfectants are recognized by the federal government. They are, in Example 1 order of increasing stringency of efficacy requirements: The following composition was prepared (amounts (1) minimal claim given are parts by weight): (2) general, and Oxine 4.1 (3) hospital type. Copper hydrate 1.4 It is obvious that a hospital-type disinfectant should be DDBSA 64.0 the most desirable for all usage, but the other two classi Propylene glycol methyl ether 30.5 fications are made necessary as a practical concession to 65 Thirty parts by weight of this composition were the economic, technical and safety problems encoun blended, at 130 F., with agitation, with: tered in having a single, most stringent only, category, Laurel alcohol ethoxylate (20 EO) 15 Hence the need for a generally applicable cleaning, Octyl phenol ethoxylate (9 EO) 15 4,766,113 91 92 Water, demineralized 30 Isopropanol 10 Example 8 This formulated concentrate was use-diluted with The following DDBSA/Cu-8-Q solution was pre water. A nylon and a wool carpet, both of which were pared by reacting copper hydrate and solubilizing the uncleaned after two years of regular service, were resulting Cu-8-Q in DDBSA and mixing in the remain shampooed and placed back in use in a damp indoor ing ingredients (amounts are parts by weight): location. Both came out of the shampoo operation clean Copper hydrate 1.70 and bright. Neither exhibited any bacterial- or fungal Oxine 4.44 caused odor after fourteen months in service. DDBSA 64.81 O Methanol 15.05 Example 2 Isopropanol 14.00 A preparation made from: This composition was diluted with a water carrier, as Oxine 2.0 parts by weight tabularized below, and tested in comparison with a Copper hydrate 0.7 parts by weight sodium tetrachlorophenate (23%) liquid concentrate, 15 also diluted in a water carrier, against organisms on DDBSA 400 parts by weight three species of green lumber-Douglas fir, Amabilis fir Isopropanol 300 parts by weight and Ponderosa pine. The organisms were: Distilled water 27.3 parts by weight Cephaloascus fragans a brown mold that infects cer was diluted 1:100 with tap water, and strips of alumi tain wood species num foil, lightweight cotton cloth and dry, smooth-sur 20 Trichoderma virgatum a common mold faced southern yellow pine wood were dipped for three Mixed spores a combination of two molds (Penicil minutes therein. Treated and untreated strips were al lium sp. and Aspergilis niger) and a fungus (Cerato lowed to air dry and remain exposed to the air for seven cystis pilifera that causes blue stain in wood. weeks. The strips then were tested by placing a Staphy The freshly cut wood samples were dip treated (15 lococcus aureus culture on the test surfaces with these 25 second immersion) with the test fungicides and then results: innoculated with spore suspensions of the above described fungi. The test boards plus untreated control Average Zone of boards were then placed in a warm, humid chamber for Sample Inhibition (mm) four weeks. The results are set forth in the table below 30 in which: Untreated aluminum foil O Treated aluminum foil 1.0 Untreated cotton cloth O Treated cotton cloth 2.5 Mixed Untreated pine wood O Use - C fragans T. virgatum spores Treated pine wood 5.0 35 Dilution A B C A B C A B C Tetra- 1:100 4 4 4 0 0 3 3 4 4 chlorophenol composition Examples 3-7 DDBSA/Cu-8-Q 1:240 2 0 1 3 0 4 4 3 4 solution To demonstrate the antifungal properties of various 40 Tetra- 1:50 2 0 4 0 0 1 3 2 4 metal-oxine chelates, compositions were prepared using chlorophenol 6 parts by weight of the indicated metal-oxine plus 64 composition parts DDBSA plus 31 parts methanol. These composi DDBSA/Cu-8-Q 1:120 0 0 1 3 0 2 2 2 solution tions were diluted 1:200 with a water carrier for use, Tetra- 1:25 2 2 3 0 0 1 0 O 3 and freshly cut pine boards were dip immersed in the 45 chlorophenol formulation, to be tested. The boards, along with an composition DDBSA/Cu-8-Q 1:60 0 0 0 1 0 0 0 0 0 untreated control, were placed in a chamber for 28 days solution and maintained at a temperature of about 80 F. and a Tetra- 1:12.5 0 1 0 0 0 0 0 O 2 humidity of about 70%. After the test period, the speci chlorophenol men boards were evaluated in terms of percentages of 50 composition DDBSA/ 1:30 0 0 0 0 0 0 0 0 0 total surface area covered by fungal stain and mold Cu-8-Q solution growth. Control 4 4 4 3 4 4 4 4 4 (no treatment) A = Douglas fir Example Metal-Oxine % Fungal Growth 55 B = Annabilis fir C = Ponderosa pine 3. Copper 17 0 = no growth 4 Tin 20 1 = no growth for 2 weeks 5 Aluminum 28 2 = medium growth 6 Nickel 39 3 = heavy growth in 4 weeks 7 Zinc 46 4 = heavy growth in 2 weeks 60 The foregoing metal-oxines (metal-8-quinolinolates) Example 9 also may be prepared in situ in the compositions by The following composition was tested for minimum reacting oxine with any of a number of appropriate fungicidal concentration and compared to two well metal compounds. Although the copper chelate of 65 known antifungal chemicals-pentachlorophenol and oxine generally is the most effective and versatile for a 2,3,5, trichloro-4-propyl-sulfonyl pyridine-and broad range of end uses, other metal-oxine chelates DDBSA. have utility. Oxine 8.2 parts by weight 4,766,113 93 94. Copper hydrate 2.8 parts by weight sure, and Poria monticola (Madison 698, ATCC 11538) DDBSA 59.0 parts by weight which is the standard copper-tolerant fungus for ground Propylene glycol methyl ether 300 parts by weight contact wood use. All of the test composition concentrations to be In the table below, the results are expressed as per tested were incorporated in the fungal growth media centage weight loss of the wood test blocks from decay (agar) in accordance with standard microbiological fungi attack. practices. Agar plugs containing the test fungicides then Composition A is comprised of: were inoculated with a sporulating culture and inocu Oxine 4.42 parts by weight lated at the temperatures and times specified by The Copper hydrate 1.51 parts by weight American Type Culture Collection (ATCC) recom 10 DDBSA 64.07 parts by weight mendations. The plugs were then scored for absence or Methanol 30.00 parts by weight presence of organism growth. The results are shown in The composition was diluted 1:110 in a water carrier for the following table. Minimum fungicidal concentrations impregnation of the L. trabea test blocks and 1:55 for were determined against a broad spectrum of fungi that test against P. monticola, are detrimental to man, foodstuffs and materials and 15 which can result in metabolite formations (mycotoxins) Composition B was the same as Composition A ex of extreme toxicity to man and animals. cept that it was diluted 1:55 with toluene carrier for In the table below, Composition A is that of this block impregnation for test against both test fungi. Example 9 and the active ingredient is Cu-8-Q; Compo Composition C was the same as the DDBSA/Cu-8-Q sition B is DDBSA; Composition C is pentachlorophe solution of Example 25 diluted with a water carrier nol; and Composition D is 2,3,5-trichloro-4-propylsul 20 1:220 for test against L. trabea and 1:55 against P. mon fonyl pyridine. Where no concentration is listed, no ticola. data is available. The stated value of '1' means 1 or Cunilate 2174 was diluted 1:110 with a toluene car fewer ppm. re. The abbreviation "pcf' means pounds of Cu-8-Q per 25 cubic foot of wood. Minimum Fungicidal Concentration - (ppm of active ingredient) - A B C D Lenzites trabea Poria nonticola Aspergillis niger 1 10,000 1-3 - Retention Weight Retention Weight (ATCC 9642) 30 Composition (pcf) Loss (%) (pcf) Loss Aspergillis terreus 1 100 - 36 A 0,021 0.3 0.040 1.3 (ATCC 10609) B 0.025 5.0 0.029 11.3 Aspergilis flavus 1 1,000 22-54 - C 0.018 2.0 0.036 8. (ATCC 11655) Cunilate 2174 0.024 13.6 0.026 43.4 Alternaria alternata 1 - - - Untreated control O 45.3 O 55.1 (ATCC 13963) 35 Aureobasidium pullulans 1 100 - -- (ATCC 16624) Lenzites trabea 100 1-3 - These results demonstrate the efficacy of the compo (ATCC 11539) sitions of the invention in both a water and an organic Polyporus tulipiferae 1. 100 1-3 -- solvent (toluene) carrier. Also illustrated is the greatly (ATCC 11245) Penicillium brevi 1 100 - - improved efficacy over Cunilate 2174, especially for compactum (ATCC 16024) wood in ground contact service. The wood protection Rhizopus stolonifer 1 100 1-3 --- results with Compositions A, B and C compare favor (ATCC 24794) ably with those of pentachlorophenol (PCP), tested Trichoderna viride 10 100 - - (ATCC 8678) simultaneously, wherein PCP, a world standard for Trichoderma sp. 1 100 - - 45 wood preservation, exhibited 1.4% weight loss at 0.30 (ATCC 12668) pcfretention against L. trabea and 3.1% weight loss at Candida albicans 1,000 - 3 0.27 pcf retention in the wood against P. monticola. (ATCC 10259) Employing AWPA Test Method M12-72 (revised 1973) for testing wood block resistance to termite These results illustrate the high efficacy of the com 50 (Reticuliterimes flavipes) attack, it was determined that position of this invention and confirm the fact that an no attack occurred at retentions of Composition A ade antifungal composition prepared from a Cu-8- quate to protect the wood from decay. Q/DDBSA solution is much superior to DDBSA alone. The results also indicate the favorable relative efficacy Example 11 of the test composition compared to the two commer 55 The purpose of this test was to determine antifungal cially available fungicides of recognized high perfor efficacy of DDBSA/Cu-8-Q solutions and to compare aC. their efficacy to that of a world standard, sodium penta chlorophenate, and a mixture of two well-known agri Example 10 cultural fungicides, Topsin M (a thiophenate) and Test compositions were evaluated as wood preserva Nabam (a thiocarbomate). The test method is desig tives via a standard soil block culture procedure nated as a proposal for the Finnish NWPC Standard (ASTM D4131-61) wherein the wood blocks were wa No. 1.4.1.3/1974. The test substrate was fresh cut, green ter-leached in accordance with standard technique pine wood. The test fungi were: prior to exposure to the test fungus. The test fungi were Blue stain fungi-mixture of Ceratocystis pilifera Z11 those specified for wood decay evaluation by the Amer 65 Sclerophoma entoxylina Z1; Pullularia pullulans U2 ican Wood Preservers' Association (AWPA)-namely, Mold fungi-mixture of Paecilomyces varioti X15 Lenzites trabea (Madison 617, ATCC 11539) which is Cladosporium sphaerospermum R7 Aspergillis ann the standard test fungus for above-ground wood expo stelodami X19 4,766,113 95 96 In the table below, Composition A is the formulation strates, and Trichophyton mentagrophytes, a cause of of Example 3. 'athlete's foot.' Composition B is the formulation of Example 3 ex A. niger-at 1:200 use dilution in a water carrier, no cept that the dilution with a water carrier was 1:100 growth after 10 minutes' exposure. rather than 1:200. T. mentagrophytes-at 1:750 use dilution in a water Composition C is a 1.5% concentration of sodium carrier, no growth after 10 minutes' exposure. pentachlorophenate in water. Simular but somewhat lower efficacy results were Composition D is a 0.4% concentration in water of a obtained by substituting zinc-8-Q or aluminum-8-Q in 47:53 weight ratio of Topsin M:Nabam. the composition of this example, produced by reacting E refers to untreated control pine boards. 10 zinc oxide and aluminum hydroxide respectively with The rating index is: OXe. 0=no visible growth l=traces of growth Example 13 2=slight growth The DDBSA/Cu-8-Q solution of Example 9 was 3=moderate growth 15 tested, along with a number of well known antimicro 4= covered with fungi bial agents, against a broad spectrum screen of economi cally important Gram-positive and Gram-negative bac teria. All antimicrobial agents were incorporated in the Test Composition agar bacterial growth media according to standard mi A. B C D E 20 crobiological practices. The bacterial species were Blue stain fungi growth 0.8 0.3 0.5 2.5 3.9 grown in nutrient broth; 24-hour cultures, the inocu Mold fungi growth 0.5 0 0.3 1.3 3.6 lum, then were streaked onto the nutrient agar plates containing the test antimicrobials. After a 24-hour incu The demonstrated efficacy of Compositions A and B bation at the appropriate temperature, the plates were against the six listed fungal organisms has utility not 25 rated for presence or absence of bacterial growth. only on the tested substrate-wood-but also for pro Minimum bactericidal concentrations for each of the tection of a variety of other materials that are attacked tested agents are stated in the following tabulation of by one or more of the fungi, including paint, concrete, results in parts per million (ppm) of active ingredient as brick, textiles and leather. defined in the description of each agent.

Antimicrobial Agent (ppm of active ingredient) Bacteria (ATCC No.) A. B C D E F G

Bacillus cereus 1 100 - 7 8 5-10 m Bacillus lichenforms (27326) l 100 - 7 8 2-5 m Bacillus megaterium (27327) 1. 100 - 7 8 - - Bacillus subtilis (37328) l 100 - 750 8 - 3 Micrococcus flavus (10240) 1 100 - 7 8 - o Mycobacterium phlei (15610) 10 - 7 8 - 3 Staphylococcus aureus (6538) 1 100 2083 7 8 1-3 3

Alcaligenes faecalis (337) O 1000 -- 750 80 - --- Alcaligenes marshali (21030) 104 100 - 7 8 - --- Esherichia coli (11229) 104 10,000 -- 750 80 250-500 165 Flavobacterium arboresceus (4358) 10 10 4166 7 8 - --- Klebsiella pneumoniae (4356) 10 10,000 - 750 8 - m Proteus vulgaris 10 1000 -- 750 800 - m Pseudomonas aeruginosa (15442) 104 000 466 750 800 1000-2500 165 Salmonella cholerasuis (10708) 104 1000 - 750 80 250-500 165 Salmonella typhi (6539) 104 1000 2083 750 80 - 165 'stated value of 'i' means 1 or less A = the DDBSA/Cu-8-Q solution of Example 9 with the active ingredient expressed in terms of Cu-8-Q. B = odBSA C = phenol D = Betadine, an iodine/polyvinylpyrrollidone complex containing 0.75% iodine. The active ingredient is iodine. E = Alkyl dimethyl ammonium chlorides (61% C, 23%. C4, 11% C6 and 3% C10). F = sodium pentachlorophenate G = 2,3,5-trichloro-4-propylsulfonyl pyridine. Example 12 These data demonstrate the high efficacy of the com Using the standard AOAC fungicidal test method position of Example 9. On the basis of the average of the (12th Edition, 1975), the composition below was evalu 60 efficacies against all the test organisms, Composition A ated against two widespread fungi. is 45 times superior to Composition B; 88 times better Oxine 2.08 parts by weight than Composition C; 10.7 times better than Composition Copper hydrate 0.70 parts by weight D; and 3.5 times superior to Composition E. DDBSA 40.00 parts by weight On the basis of the average of the efficacies against Isopropanol 32.00 parts by weight 65 the three test bacteria (Staphylococcus aureus, Salmo Demineralized water 25.22 parts by weight nella cholerasuis and Pseudomonas aeruginosa PRD-10) The two fungi were Aspergillis niger, a ubiquitous required by the Environmental Protection Agency of a black fungus which fluorishes on a broad range of sub "hospital grade' disinfectant, Composition A is 10 times 4,766,113 97 98 better than Composition B; 7.2 times better than Com position D; and 1.6 times better than Composition E. Ring Rot % % % Yield Example 14 Contam- Plant Ring Rot Ring Rot cwt/ Antimicrobial inated Stored Plants Tibers acre The composition set forth below was prepared by None (control) Yes 98 23 8 493 previously described procedures: None (control) No 95 O O 609 Oxine 2.08 parts by weight DDBSA/ Yes 98 O 2 631 Copper hydrate 0.70 parts by weight Cu-8-Q DDBSA No. 100 O O 602 Isopropanol 32.00 parts by weight Cu-8-Q DDBSA 40.00 parts by weight 10 20% Clorox Yes 98 20 9 500 Water (demineralized) 25.22 parts by weight 20% Clorox No 98 O O 602 When evaluated as a bactericide by the AOAC Use Roccal Yes 98 20 9 515 Dilution Method (12th Edition, 1975), 10 ring carriers Roccal No 98 O O 638 per organism, the following results were obtained (A= - subculture and B=resubculture): 15 The composition of this example demonstrates supe rior control of the ring rot bacterium. Other species of the genus Corynebacterium are causal agents of disease Use Dilution Negative - Positive - in man and a variety of plant life. In Water Carrier A. B. A B The above examples illustrate the basic high order Staphylococcus aureus 1:1000 O 10 O O 20 inhibition characteristics of the disclosed compositions Salmonella cholerasuis 1:1000 O 10 O O against a range of odor-producing fungi and bacteria. (PRD-10) Pseudomonas aeruginosa 1:400 0 10 0 O We claim: Aerobacter aerogenes 1:400 O 10 0 O 1. An antimicrobial composition for controlling bac teria and fungi comprising in combination: 25 (a) a metal chelate of 8-hydroxy quinoline in which A ten-minute kill is required against the first three the metal is selected from the group consisting of pathogens for sale as a hospital-grade disinfectant. Effi copper, mercury, cadmium, nickel, tin, aluminum cacy against the fourth organism, a major cause of slime and zinc, in recirculated cooling water systems and pulp and (b) an alkyl benzene sulfonic acid wherein the alkyl paper mills, demonstrates utility of the composition as a 30 group is C6 to C18, said metal chelate being solubi slimicide. lized by said sulfonic acid, Example 15 2. A composition according to claim 1 wherein the composition includes 1-50 parts by weight of a polar This composition was prepared and tested at one use diluent. dilution, 1:50 in water carrier, against the causal agent 35 3. A composition according to claim 2 wherein said of potato ring rot bacteria (Corynebacterium Sepedoni polar diluent is methanol, ethanol, isopropanol, n cum): butanol, dimethylformamide, N-methyl-2-pyrrollidone, Oxine 2.08 parts by weight ethylene glycol or water. Copper hydrate 0.70 parts by weight 4. A composition according to claim 1 wherein said DDBSA 40.00 parts by weight metal chelate is copper-8-quinolinolate. Triton X-100 20.00 parts by weight 5. A composition according to claim 2 wherein said Isopropanol 22.00 parts by weight metal chelate is copper-8-quinolinolate. Water (demineralized) 15.22 parts by weight 6. A composition according to claim 1 wherein said Infected potato seed readily contaminate potato seed sulfonic acid is dodecylbenzene sulfonic acid. cutters, sacks, bins, cellars, trucks and planting equip 45 7. A composition according to claim 2 wherein said ment with the highly infectious ring rot bacteria. The sulfonic acid is dodecylbenzene sulfonic acid. result may be infected potato plants, tubers and reduced 8. A composition according to claim 3 wherein said yields. sulfonic acid is dodecylbenzene sulfonic acid. The test procedure consisted of dipping unpainted, 9. A composition according to claim 4 wherein said planed wood laths (6') into a slurry of infected ring rot 50 sulfonic acid is dodecylbenzene sulfonic acid. tuber tissue, allowing excess slurry to drain off (3-5 10. A method for controlling the growth of microor minutes) and then spraying the contaminated lath with ganisms which comprises applying to the locus of said the test antibacterials. Three to five minutes later, microorganisms an antimicrobially effective amount of healthy Norgold Russet potato seed pieces were rubbed a composition comprising: vigorously against both sides of the contaminated and 55 (a) a metal chelate of 8-hydroxy quinoline in which antibacterial-treated laths. The process was repeated the metal is selected from the group consisting of using laths not contaminated with C. sepedonicum but copper, mercury, cadmium, nickel, tin, aluminum treated with the test antibacterial agent. The rubbed and zinc, seed pieces were stored in bags and later planted at the (b) an alkyl benzene sulfonic acid wherein the alkyl appropriate time. 60 group is C6 to C18, said metal chelate being solubi In addition to the composition of the invention, un lized by said sulfonic acid, treated controls, 20% Clorox (1.05% sodium hypochlo 11. A method according to claim 10 wherein said rite in water), formaldehyde (37% formaline diluted composition includes 1-50 parts by weight of a polar 1:120 in water) and Roccal (benzalkonium chloride or diluent. Zephiran chloride) diluted with water to 800 ppm con 65 12. A method according to claim 11 wherein said centration were tested. The results of the test are tabu polar diluent is methanol, ethanol, isopropanol, n lated below and refer to plants and tubers produced butanol, dimethylformamide, N-methyl-2-pyrrollidone, from the tubbed test seed pieces. ethylene glycol or water. 4,766,113 99 100 16. A method according to claim 10 wherein said 13. A method according to claim 10 wherein said sulfonic acid is dodecylbenzene sulfonic acid. metal chelate is copper-8-quinolinolate. 17. A method according to claim 11 wherein said sulfonic acid is dodecylbenzene sulfonic acid. 14. A method according to claim 11 wherein said 5 18. A method according to claim 12 wherein said metal chelate is copper-8-quinolinolate. sulfonic acid is dodecylbenzene sulfonic acid. 19. A method according to claim 13 wherein said 15. A method according to claim 12 wherein said sulfonic acid is dodecylbenzene sulfonic acid. metal chelate is copper-8-quinolinolate. k sk k 10

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