Abstract:
This invention relates to a reaction product of allantoin and formaldehyde made in a molar ratio of about 1:2.75–1:3.25, preferably 1:3, respectively, preferably under controlled pH (5.0 to 7.0) and temperature (40° to 85° C., preferably 50–60° C.) conditions, which product contains free formaldehyde (&lt;0.1%), and methylene diol of about 580–1385 ppm, preferably at a pH of 7.2, with advantageous long-lasting, anti-microbial properties, particularly against the organism  B. cepacia , and in combination products with parabens and iodopropynyl butyl carbamate (IPBC).

Description:
CROSS REFERENCE TO RELATED APPLICATION 
     The present application is a continuation-in-part of application Ser. No. 10/302,093, filed Nov. 22, 2002, now abandoned. 
    
    
     BACKGROUND OF THE INVENTION 
     1. Field of the Invention 
     This invention relates to reaction products of allantoin and formaldehyde and, more particularly, to such products which contain very low levels of free formaldehyde (&lt;0.1%), while retaining advantageous, long-lasting anti-microbial properties, particularly against the organism  B. cepacia.    
     2. Description of the Prior Art 
     Berke, P. A., in U.S. Pat. No. 3,248,285 described a reaction product of allantoin and formaldehyde (Germall® 115) from the reactants in a mole ratio of 1:1.5, respectively; and in U.S. Pat. No. 4,271,176 and Reissue 32,848, (Germall® II) in a mole ratio of 1:4. However, during recent evaluations using modern C 13  NMR techniques, these reaction products were found to contain 0.5% to 1.0% of free formaldehyde. This amount was considered in the past to be necessary to retain its anti-microbial activity; although now it is recognized that a considerable amount of free formaldehyde in the product is disadvantageous from a safety (irritation) and environmental standpoints. 
     Accordingly, it is an object of this invention to prepare reaction products of allantoin and formaldehyde which have long-lasting anti-microbial properties with the presence therein of only very low levels of free formaldehyde (i.e. &lt;0.1%). 
     Another object of the invention is to provide a method of making such effective reaction products. 
     Still another object of this invention is to provide a synergistic combination of such products with other known fungicides to obtain broad spectrum activity against bacteria and fungi, yeast, molds, and the like. 
     SUMMARY OF THE INVENTION 
     This invention relates to a reaction product of allantoin and formaldehyde, 
                                
which is
 
                                
or mixtures thereof, made in a molar ratio of about 1:2.75–1:3.25, preferably 1:3, respectively, preferably under controlled pH (5.0 to 7.0) and temperature (40° to 60° C.) conditions, which contains substantially no free formaldehyde (&lt;0.1%), with advantageous long-lasting, broad spectrum anti-microbial properties, particularly against the organism  B. cepacia.    
     What has been discovered herein is that reaction products of allantoin and formaldehyde result in free formaldehyde being present in equilibrium with methylene diol and N-methylol. Unexpectedly, it was discovered that N-methylol and methylene diol, work in synergy to act as long-lasting anti-microbial agents, i.e. both can slowly react to provide anti-microbial protection against a wide range of microbes, including the difficult to kill  B. cepacia , while free formaldehyde is present in very low concentrations (&lt;0.1%) in equilibrium with methylene diol. 
     
       
                 
         
             
             
         
      
     
     Both equilibrium moieties can react with the amide moieties of allantoin to form several N-methylol of the allantoin compounds, which are stabilized by H-bonding; however they can slow hydrolyzed in water to give the methylene diol intermediate, which can react with amide to form N-methylols. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The following examples describe preparation of Comparative Reaction Products (CRP) for (A) Germall® 115 (1:1.5); at low caustic levels; and (B) at high caustic levels; and (C) Germall® II (1:4), low caustic and (D) high caustic; and Invention Reaction Products (IRP) Germall® III (1:3), (A) laboratory and (B) commercial runs, with only enough caustic to neutralize the formic acid present in the Formalin solution. 
     Comparative Reaction Products 
     
       
         
               
             
               
               
               
             
           
               
                   
               
               
                 A. GERMALL ® 115 
               
               
                 (1:1.5) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Allantoin 
                 15.8 g (0.1 mole) 
               
               
                   
                 Formalin (37%) 
                 12.2 g (0.15 mole) 
               
               
                   
                 Water 
                 28.5 ml 
               
               
                   
                   
               
             
          
         
       
     
     The above mixture was refluxed for one hour to form a clear solution. 
     
       
         
               
             
               
               
               
             
           
               
                   
               
               
                 B. GERMALL ® 115 (High Caustic) 
               
               
                 (1:1.5) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Allantoin 
                 600 g (3.8 mole) 
               
               
                   
                 Formalin (37%) 
                 450 g (5.5 mole) 
               
               
                   
                 Sodium Hydroxide 
                 123 g 
               
               
                   
                   
               
             
          
         
       
     
     Refluxed for one hour to form a clear solution. Concentrated acetic acid was added to adjust the Ph to 4.0. Removed water to give a white powder. 
     
       
         
               
             
               
               
               
             
           
               
                   
               
               
                 C. GERMALL ® II 
               
               
                 (1:4) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Allantoin 
                 1053 g (6.66 mole) 
               
               
                   
                 Formalin (37%) 
                 2160 g (26.64 mole) 
               
               
                   
                   
               
             
          
         
       
     
     The white suspension was heated to 85° C. and held for an additional hour; upon cooling a clear solution was obtained. Removed water under reduced pressure to give a white powder. 
     
       
         
               
             
               
               
               
             
           
               
                   
               
               
                 D. GERMALL ® II 
               
               
                 (1:4) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Allantoin 
                 158.1 g (1.0 mole) 
               
               
                   
                 Formalin (37%) 
                 324.2 g (3.99 mole) 
               
               
                   
                 Sodium Hydroxide 10% 
                  32.0 g (0.08 mole) 
               
               
                   
                   
               
             
          
         
       
     
     Refluxed at 85° C. for one hour. The clear colorless solution obtained was dried under reduced pressure to give solid white powder residue. 
     Comparative Test Results 
     Activity against yeast and mold: 
     0.3% test solutions. 
     
       
         
               
               
               
             
           
               
                   
               
               
                   
                 
                   C. Albican 
                 
                 
                   A. Niger 
                 
               
               
                 TEST SOLUTIONS 
                 ATTC1023 
                 ATCC 9642(for 3 Days) 
               
               
                   
               
             
             
               
                 0.3% Germall ® 115 (Exs. A/B) 
                 + 
                 + 
               
               
                 0.3% Germall ® II(Exs. C/D) 
                 − 
                 − 
               
               
                   
               
               
                 + Growth 
               
               
                 − No Growth 
               
             
          
         
       
     
     Invention Reaction Product 
     
       
         
               
             
               
               
               
             
           
               
                   
               
               
                 A. (LABORATORY SCALE) (GERMALL ® III) 
               
               
                 (1:3) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Allantoin 
                 1616 g (10.23 mole) 
               
               
                   
                 Formalin LM*(37%) 
                 2488 g (30.68 mole) 
               
               
                   
                 Sodium Hydroxide 50% 
                  24 g 
               
               
                   
                   
               
               
                   
                 *LM = low methanol (&lt;0.5%) Borden Chemicals 
               
             
          
         
       
     
     Mixed and heated at 60° C. for 3 hours to give a clear solution. The Ph of the product was adjusted to 7.2 with the sodium hydroxide solution to neutralize formic acid in Formalin® and the solution was spray dried to give a free-flowing, white powder. 
     Invention Reaction Product 
     
       
         
               
             
               
               
               
             
           
               
                   
               
               
                 B. (COMMERCIAL SCALE) (GERMALL ® III) 
               
               
                 (1:3) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Allantoin.wet cake 
                 2095 lbs (10.23 mole) 
               
               
                   
                 Formalin LM (37%) 
                 2488 lbs (30.68 mole) 
               
               
                   
                 Sodium Hydroxide 50% 
                  23.6 lbs 
               
               
                   
                 Ph 
                 6.5–7.0 
               
               
                   
                 Reaction Temp 
                  40–60° C. 
               
               
                   
                   
               
             
          
         
       
     
     The resultant mixture then was further reacted at 85° C. for 3 hours to give a clear solution at pH 7.2. The solution was spray dried to remove water and other volatile by-products to give a free-flowing, white powder. 
     A study was conducted to determine the level of methylene diol in the reaction products versus the number of equivalents of formaldehyde added during formation. These results are based on quantitative 13C-NMR analysis and summarized in Table 1 below. 
     
       
         
               
               
               
             
               
               
               
             
           
               
                   
                 TABLE 1 
               
               
                   
                   
               
               
                   
                 Number of Equivalents 
                   
               
               
                   
                 Formaldehyde/Allantoin 
                 ppm Methylene Diol 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 4.00 
                 3545 
               
               
                   
                 3.50 
                 1916 
               
               
                   
                 3.25 
                 1385 
               
               
                   
                 3.00 
                 790 
               
               
                   
                 2.75 
                 583 
               
               
                   
                 2.50 
                 340 
               
               
                   
                 1.50 
                 250 
               
               
                   
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 BIOACTIVITY OF GERMALL ® COMPOUNDS 
               
             
          
           
               
                 INVENTION 
                   
                   
                   
               
               
                 EXS. PRESERVATIVE 
                 ORGANISM 
                 STATIC 
                 CIDAL 
               
               
                   
               
               
                 IRP - Germall ® III (1:3) 
                 
                   Staph aureus 
                 
                  300 ppm 
                 1250 ppm 
               
               
                   
                 
                   E. coli 
                 
                  300 ppm 
                 1250 ppm 
               
               
                   
                 
                   P. aeruginosa 
                 
                  300 ppm 
                  600 ppm 
               
               
                   
                 
                   B. cepacia 
                 
                  150 ppm 
                  300 ppm 
               
               
                   
                 
                   C. albicans 
                 
                 &gt;5000 ppm   
                 — 
               
               
                   
                 
                   A. niger 
                 
                 2500 ppm 
                 2500 ppm 
               
               
                 CRP - Germall ® II (1:4) 
                 
                   Staph aureus 
                 
                  300 ppm 
                 1250 ppm 
               
               
                   
                 
                   E. coli 
                 
                  600 ppm 
                 1250 ppm 
               
               
                   
                 
                   P. aeruginosa 
                 
                  600 ppm 
                 1250 ppm 
               
               
                   
                 
                   B. cepacia 
                 
                  150 ppm 
                  600 ppm 
               
               
                   
                 
                   C. albicans 
                 
                 5000 ppm 
                 &gt;5000 ppm   
               
               
                   
                 
                   A. niger 
                 
                 2500 ppm 
                 2500 ppm 
               
               
                 CRP - Germall ® 115 
                 
                   Staph aureus 
                 
                 1250 ppm 
                 2500 ppm 
               
               
                 (1:1.5) 
                 
                   E. coli 
                 
                 1250 ppm 
                 2500 ppm 
               
               
                   
                 
                   P. aeruginosa 
                 
                 1250 ppm 
                 2500 ppm 
               
               
                   
                 
                   B. cepacia 
                 
                  600 ppm 
                 1250 ppm 
               
               
                   
                 
                   C. albicans 
                 
                 &gt;5000 ppm   
               
               
                   
                 
                   A. niger 
                 
                 5000 ppm 
                 5000 ppm 
               
               
                   
               
             
          
         
       
     
     Protocol 
     Minimum Inhibitory Concentration (MIC) Test Method 
     Scope 
     The purpose of this test procedure is to screen experimental compounds for anti-microbial activity. 
     Principle 
     The measurement of the lowest effective concentration of an anti-microbial or anti-microbial blend is important for recommending use concentrations. The MIC test is an in vitro tube dilution procedure used to identify effective concentrations of anti-microbials. In this test, the experimental compound is diluted by serial concentrations into nutrient culture media. Test organisms are then inoculated into the anti-microbial solutions. 
     If the experimental compound is effective, there is no growth observed in the test dilution tubes and they are clear. If the experimental compound is not effective, the test dilution tubes are cloudy, indicating growth. This test will determine static as well as cidal activity concentrations. 
     Materials 
     
         
         1. Laminar flow hood (Baker Sterilgard SG 400) 
         2. 18×150 mm culture tubes 
         3. Stock antimicrobial solution 
         4. Media: Trypticase soy broth (BBL 11043) and AOAC Letheen broth (BBL 10914) 
         5. Test organisms:  Staphylococcus aureus  ATCC 6538,  Escherichia coli  ATCC 8739,  Pseudomonas aeruginosa  ATCC 9027,  Burkholderia cepacia  ATCC 25416,  Candida albicans  ATCC 10231, and  Aspergillus niger  ATCC 16404. 
         6. Spectrophotometer (Spectronic 20D, Milton Roy) 
       
    
     Minimum Inhibitory Concentration Test 
     Procedure 
     
         
         1. Antimicrobial stock solutions are prepared at predetermined concentrations (i.e., 10% through 0.07%) depending on the test material. Serial doubling dilutions are made as follows. Each culture tube contains 5 mls of trypticase soy broth. Five mls of the stock solution are added to the first tube and vortexed. 5 mls are then removed and placed into the second tube, (and so on, until the last tube). At the final test concentration, 5 mls of the broth/antimicrobial mixture is decanted out. 
         2. The test organisms are prepared as with any organism inoculum (MLM 100-3, MLM 100-4, and MLM 100-5). A saline suspension of each organism is prepared. The bacterial organisms and the yeast are a standardized at a concentration of 1×10 6  cfu/ml. The mold inoculum is approximately 1×10 5  cfu/ml. 
         3. Inoculate each culture tube with 0.10 mls of organism inoculum and vortex. 
         4. Incubate bacterial tubes for 24 hours at 35° C. Incubate yeast or mold tubes for 48 hours at 25° C. Read for growth; turbid tubes for bacteria and yeast; mold clearly visible tubes. This is the minimum inhibitory concentration (static activity). 
         5. After the tubes are read, transfer all “clear” tubes and the first cloudy (growth) tube into Letheen broth containing neutralizers. Incubate the Letheen broth tubes for 48 hours at the bacterial or fungal incubation temperatures. Read for growth; turbid tubes for bacteria and yeast; mold clearly visible in mold tubes. This is the cidal activity concentration.
 
Discussion
 
       
    
     The cidal activity of an anti-microbial can be rapidly screened by means of a MIC test before further evaluation tests, such as longer preservative efficacy tests, are performed. This test is a tube serial dilution procedure limited only by the water solubility of the material. Where anti-microbial materials are slightly insoluble, leaving the TSB broth turbid, a procedure modification can be made. Tubes are incubated for 24 hours (bacteria) or 48 hours (fungi) but instead of transfer to Letheen broth, the TSB tubes are streaked onto Letheen agar. The agar plates are then incubated appropriately and then read for absence or presence of growth. Depending on the degree of insolubility, a measure of cidal activity may be the only parameter measured. 
     Anti-microbial neutralization is important in this screening test. Letheen broth or agar contains neutralizers but if these do not neutralize the anti-microbial adequately, others can be added. These are to be determined prior to testing. 
     Aseptic technique is important in any microbiological procedure. All functional operations are performed under the laminar flow hood with use of sterile pipettes, tubes and media to eliminate cross-contamination. Surface sanitizers (i.e., alcohol) are used on the work surface before and after each operation. Ample time is allowed for recirculation of air within the sterile chamber of the hood. 
     The bioactivity data show particular effectiveness against the organism cepacia B. (cidal=300 ppm vs. 600 ppm and 1250 ppm for Germall® II and Germall® 115, respectively). However, if desired, even broader spectrum antibacterial activity can be achieved by combination products with the invention composition whose formulations are given below. 
     Invention Combination Products 
     
       
         
               
             
               
               
               
             
           
               
                   
               
               
                 COMBINATION BLENDS (BY WEIGHT) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 (1) 
                 Germall ® III 
                 20–30% 
               
               
                   
                 MP - methyl paraben 
                  8–12% 
               
               
                   
                 PP - propyl paraben 
                 2–4% 
               
               
                   
                 PG - propylene glycol 
                 q.s. 100 
               
               
                 (2) 
                 Germall ® III 
                 40–45% 
               
               
                   
                 IPBC - iodopropynyl butyl carbamate 
                 0.5–5%   
               
               
                   
                 PG - propylene glycol 
                 qs 100 
               
               
                 (3) 
                 Germall ® III 
                 98.5–99.5% 
               
               
                   
                 IPBC - iodopropynyl butyl carbamate 
                 0.5–1.5% 
               
               
                   
                 (Powder) 
               
               
                   
               
             
          
         
       
     
     Preservative Activity (Challenge Test) 
     A typical cosmetic emulsion was prepared for microbiological challenge testing and predetermined admixtures of a methylol compound and IPBC were added at various use levels. The emulsion thus prepared had the following composition: 
     
       
         
               
             
               
               
               
             
               
               
               
             
           
               
                   
               
               
                 NONIONIC EMULSION (UNPRESERVED CONTROL) 
               
             
          
           
               
                 Phase 
                 Ingredient 
                 % wt. 
               
               
                   
               
             
          
           
               
                 A 
                 Water 
                 69.80 
               
               
                 A 
                 Carbomer 
                 10.00 
               
               
                 B 
                 Octyl Palmitate 
                 5.00 
               
               
                 B 
                 Cetearyl alcohol and Ceteareth-20 
                 2.00 
               
               
                 B 
                 Glyceryl Stearate and Laureth-23 
                 2.50 
               
               
                 B 
                 Mineral Oil 
                 5.00 
               
               
                 C 
                 Triethanolamine (99%) 
                 0.20 
               
               
                 D 
                 Preservative 
                 0.00 
               
               
                 E 
                 Hydrolyzed Collagen 
                 0.50 
               
               
                 E 
                 Water 
                 5.00 
               
               
                   
                 Total 
                 100.00 
               
               
                   
               
               
                 Procedure: 
               
               
                 Heat Phase A to 75° C. Heat Phase B to 75° C. 
               
               
                 Add Phase B to Phase A. Mix until uniform. 
               
               
                 Add Phase C. Remove heat. 
               
               
                 Add Phase D at the appropriate temperature. 
               
               
                 Add Phase E at 40° C. 
               
               
                 Continue mixing to 30° C. 
               
             
          
         
       
     
     
       
         
               
             
               
               
             
               
               
               
             
           
               
                   
               
               
                 STANDARD SCREENING EMULSIONS 
               
             
          
           
               
                   
                 % wt. 
               
               
                   
                   
               
             
          
           
               
                   
                 Phase A 
                   
               
               
                   
                 Stearic Acid 
                 5.00 
               
               
                   
                 Mineral Oil 
                 2.50 
               
               
                   
                 Cetyl Alcohol 
                 1.00 
               
               
                   
                 Lareth-5 and Ceteth-5 and 
                 0.50 
               
               
                   
                 Oleth-5 and Steareth-5 
               
               
                   
                 Glycerol Monostearate and 
                 1.50 
               
               
                   
                 Polyoxyethylene Stearate 
               
               
                   
                 Phase B 
               
               
                   
                 Deionized Water 
                 88.0 
               
               
                   
                 Triethanolamine 99% 
                 1.00 
               
               
                   
                 Citric Acid 30% aqueous solution 
                 0.60 
               
               
                   
                 Preservative Admixture 
                 qs 
               
               
                   
                   
               
             
          
         
       
     
     To prepare the emulsion, Phases A and B were heated separately to 75°–80° C. Phase A then was added to Phase B with mixing. The mixture then was cooled to 55°–60° C. At this point the desired amount of the preservative admixture was added and the product was cooled to 50° C. while stirring. The citric acid solution then was added to adjust the pH and the mixture was stirred until a temperature of 30° C. was reached. 
     The challenge tests were carried out using the following microorganisms: SA, ECOLI, PSA, PC, AN and CAN, in this manner. 50 g aliquots of the test emulsion containing various amounts of the preservative admixture were inoculated with approximately 10 7 –10 8  of the challenge organisms. The test samples then were stirred to disperse the challenge inoculum. The samples were incubated and assayed at 48 hours, 7, 14, 21 and 28 days. The assays were performed on 1 g of the test sample by serially diluting 10 1  to 10 6  of the original concentration. The plating medium for bacteria was Letheen agar and for fungi it was low pH Mycophil agar with Tween 20. Each plated sample was incubated for 48 hours at 37° C. for bacteria, 5 days at 25° C. for mold, and 3 days at 25° C. for fungi. After incubation, readings of the number of colonies per milliliter (cfu/ml) were made. At 21 days the test product was reinoculated with half of the original inoculum. The data is presented in Tables 3–11 below. 
     Challenge Tests 
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 COMPARISON OF ACTIVITY OF GERMALL III TO GERMALL II AND 115 
               
               
                 (SCREENING EMULSION) 
               
             
          
           
               
                 Preservative 
                 Conc. 
                 Organism 
                 48 hrs 
                 7 days 
                 14 days 
                 21 days 
                 28 days 
               
               
                   
               
             
          
           
               
                 Germall II 
                 1000 ppm 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 160,000 
                 78,000 
                 63,000 
                 260,000 
                 210,000 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall III 
                 1000 ppm 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 160,000 
                 380,000 
                 380,000 
                 810,000 
                 640,000 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall 115 
                 2000 ppm 
                 SA 
                 3000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 490 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 230,000 
                 2,000,000 
                 650,000 
                 1,500,000 
                 1,200,000 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall II 
                 2000 ppm 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 150 
                 9,600 
                 48,900 
                 490,000 
                 210,000 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall III 
                 2000 ppm 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 6000 
                 195,000 
                 460,000 
                 690,000 
                 1,070,000 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Unpreserved 
                   0 
                 SA 
                 2,100,000 
                 57,000 
                 90 
                 &lt;10 
                 68,000 
               
               
                   
                   
                 EC 
                 37,000 
                 96,000 
                 96,000 
                 43,000 
                 790,000 
               
               
                   
                   
                 PSA 
                 70 
                 4600 
                 500 
                 10,100 
                 170,000 
               
               
                   
                   
                 BC 
                 2,100,000 
                 860,000 
                 1,520,000 
                 3,520,000 
                 &gt;10 E6 
               
               
                   
                   
                 CAN 
                 1,100,000 
                 168,000 
                 67,000 
                 270,000 
                 460,000 
               
               
                   
                   
                 AN 
                 700,000 
                 56,000 
                 44,000 
                 190,000 
                 320,000 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 COMPARISON OF ACTIVITY OF GERMALL III TO GERMALL II AND 115 
               
               
                 (NONIONIC EMULSION) 
               
             
          
           
               
                 Preservative 
                 Conc. 
                 Organism 
                 48 hrs 
                 7 days 
                 14 days 
                 21 days 
                 28 days 
               
               
                   
               
             
          
           
               
                 Germall III 
                 2000 ppm 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 820,000 
                 1,680,000 
                 1,350,000 
                 700,000 
                 &gt;1E6 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall II 
                 2000 ppm 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 320,000 
                 720,000 
                 650,000 
                 730,000 
                 &gt;1E6 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall 115 
                 2000 ppm 
                 SA 
                 1,500 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 52,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 &gt;1E6 
                 &gt;1E6 
                 &gt;1E6 
                 700,000 
                 &gt;1E6 
               
               
                   
                   
                 AN 
                 &lt;10 
                 20 
                 390 
                 370 
                 &gt;1E4 
               
               
                 Germall III 
                 4000 ppm 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 24,000 
                 &gt;1E6 
                 &gt;1E6 
                 730,000 
                 &gt;1E6 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall II 
                 4000 ppm 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 50 
                 10,000 
                 620,000 
                 460,000 
                 &gt;1E6 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall 115 
                 4000 ppm 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 1,500 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 1,060,000 
                 1,000,000 
                 &gt;1E6 
                 &gt;1E6 
                 &gt;1E6 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Unpreserved 
                   0 
                 SA 
                 &gt;1E6 
                 6,300 
                 &gt;1E4 
                 &lt;10 
                 &gt;1E4 
               
               
                   
                   
                 EC 
                 &gt;1E6 
                 900,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 PSA 
                 20,000 
                 &gt;1E6 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 BC 
                 &gt;1E6 
                 &gt;1E6 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 CAN 
                 &gt;1E6 
                 &gt;1E6 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 AN 
                 500,000 
                 510,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 COMPARISON OF ACTIVITY OF GERMALL III TO GERMALL II AND 115 
               
               
                 (SCREENING EMULSION) 
               
             
          
           
               
                 Preservative 
                 Conc. 
                 Organism 
                 48 hrs 
                 7 days 
                 14 days 
                 21 days 
                 28 days 
               
               
                   
               
             
          
           
               
                 Germall III 
                 250 ppm 
                 SA 
                 69,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 11,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 200 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 430,000 
                 120,000 
                 70,000 
                 150,000 
                 850,000 
               
               
                   
                   
                 AN 
                 100,000 
                 200 
                 70 
                 40 
                 1,300 
               
               
                 Germall II 
                 250 ppm 
                 SA 
                 55,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 5500 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 290,000 
                 170,000 
                 71,000 
                 46,000 
                 680,000 
               
               
                   
                   
                 AN 
                 50,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 100 
               
               
                 Germall 115 
                 250 ppm 
                 SA 
                 117,000 
                 30 
                 &lt;10 
                 &lt;10 
                 1,500 
               
               
                   
                   
                 EC 
                 40,000 
                 20 
                 &lt;10 
                 &lt;10 
                 3600 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 320 
                 &gt;1E4 
                 &gt;1E6 
                 &gt;1E6 
               
               
                   
                   
                 BC 
                 11,000 
                 &gt;1E6 
                 &gt;1E6 
                 &gt;1E6 
                 &gt;1E6 
               
               
                   
                   
                 CAN 
                 1,090,000 
                 270,000 
                 1,120,000 
                 770,000 
                 &gt;1E6 
               
               
                   
                   
                 AN 
                 90,000 
                 20,000 
                 20,000 
                 29,000 
                 300,000 
               
               
                 Germall III 
                 500 ppm 
                 SA 
                 38,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 18,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 100,000 
                 210,000 
                 310,000 
                 270,000 
                 &gt;1E6 
               
               
                   
                   
                 AN 
                 9000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall II 
                 500 ppm 
                 SA 
                 17,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 610 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 170,000 
                 100,000 
                 90,000 
                 320,000 
                 930,000 
               
               
                   
                   
                 AN 
                 40 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall 115 
                 500 ppm 
                 SA 
                 140,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 24,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 290,000 
                 760,000 
                 790,000 
                 1,210,000 
                 &gt;1E6 
               
               
                   
                   
                 AN 
                 130,000 
                 1,000 
                 40 
                 290 
                 80,000 
               
               
                 Unpreserved 
                  0 
                 SA 
                 &gt;1E6 
                 34,000 
                 6,800 
                 20 
                 &gt;1E4 
               
               
                   
                   
                 EC 
                 18,000 
                 4,900 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 PSA 
                 50 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 BC 
                 &gt;1E6 
                 &gt;1E6 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 CAN 
                 970,000 
                 270,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 AN 
                 150,000 
                 280,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                 COMPARISON OF ACTIVITY OF GERMALL PLUS AND GERMALL III/IPBC 
               
               
                 (SCREENING EMULSION) 
               
             
          
           
               
                 Preservative 
                 Conc. 
                 Organism 
                 48 hrs 
                 7 days 
                 14 days 
                 21 days 
                 28 days 
               
               
                   
               
             
          
           
               
                 Germall Plus 
                   
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall II 
                 1980 
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 20 
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall III 
                 1980 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 20 
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 100 
               
               
                 Germall III 
                 1960 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 40 
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Unpreserved 
                   
                 SA 
                 580,000 
                 3200 
                 180 
                 &lt;10 
                 &gt;1E4 
               
               
                   
                   
                 EC 
                 5,200 
                 70,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 PSA 
                 18,000 
                 40,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 BC 
                 &gt;1E6 
                 &gt;1E6 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 CAN 
                 &gt;1E6 
                 200,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 AN 
                 210,000 
                 270,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 7 
               
             
             
               
                   
               
               
                 COMPARISON OF ACTIVITY OF LIQUID GERMALL PLUS AND 
               
               
                 GERMALL III/IPBC-LIQ (SCREENING EMULSION) 
               
             
          
           
               
                 Preservative 
                 Conc. 
                 Organism 
                 48 hrs 
                 7 days 
                 14 days 
                 21 days 
                 28 days 
               
               
                   
               
             
          
           
               
                 LiqGermPlus 
                   
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall II 
                 790 
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 10 
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 8,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 LiqGermPlus 
                   
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall II 
                 1580 
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 20 
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 100 
               
               
                 Germall III 
                 790 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 10 
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Liquid 
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 26,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall III 
                 1580 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 20 
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Liquid 
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Unpreserved 
                 0 
                 SA 
                 580,000 
                 3200 
                 180 
                 &lt;10 
                 &gt;1E4 
               
               
                   
                   
                 EC 
                 5200 
                 70,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 PSA 
                 18,000 
                 40,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 BC 
                 &gt;1E6 
                 &gt;1E6 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 CAN 
                 &gt;1E6 
                 200,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 AN 
                 210,000 
                 270,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 8 
               
             
             
               
                   
               
               
                 COMPARISON OF ACTIVITY OF GERMALL PLUS AND GERMALL III/IPBC 
               
               
                 (SCREENING EMULSION) 
               
             
          
           
               
                 Preservative 
                 Conc. 
                 Organism 
                 48 hrs 
                 7 days 
                 14 days 
                 21 days 
                 28 days 
               
               
                   
               
             
          
           
               
                 Germall Plus 
                   
                 SA 
                 42,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall II 
                 495 
                 EC 
                 40 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 5 
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 48,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 100 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall Plus 
                   
                 SA 
                 300 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall II 
                 990 
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 10 
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall III 
                 495 
                 SA 
                 46,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 5 
                 EC 
                 25,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 11,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall III 
                 990 
                 SA 
                 24,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 10 
                 EC 
                 1,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 19,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &gt;1E6 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 2,500 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall III 
                 490 
                 SA 
                 23,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 10 
                 EC 
                 900 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 18,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &gt;1E6 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 2800 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall III 
                 980 
                 SA 
                 2,700 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 20 
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Unpreserved 
                 0 
                 SA 
                 &gt;1E6 
                 54,000 
                 4,400 
                 20 
                 &gt;1E4 
               
               
                   
                   
                 EC 
                 80,000 
                 67,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 PSA 
                 2,000 
                 4200 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 BC 
                 &gt;1E6 
                 &gt;1E6 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 CAN 
                 990,000 
                 320,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 AN 
                 380,000 
                 170,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 9 
               
             
             
               
                   
               
               
                 COMPARISON OF ACTIVITY OF LIQUID GERMALL PLUS 
               
               
                 AND GERMALL III/0.5% OR 0.8% IPBC 
               
               
                 (SCREENING EMULSION) 
               
             
          
           
               
                 Preservative 
                 Conc. 
                 Organism 
                 48 hrs 
                 7 days 
                 14 days 
                 21 days 
                 28 days 
               
               
                   
               
             
          
           
               
                 LiqGermPlus 
                   
                 SA 
                 110,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall II 
                 195 
                 EC 
                 2,600 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 2.5 
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 240,000 
                 120 
                 &lt;10 
                 &lt;10 
                 &gt;1E4 
               
               
                   
                   
                 AN 
                 230 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 LiqGermPlus 
                   
                 SA 
                 2,800 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall II 
                 390 
                 EC 
                 1100 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 5 
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 11,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 20 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 100 
               
               
                 Germall III 
                 195 
                 SA 
                 260,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 2.5 
                 EC 
                 4,300 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Liquid 
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 150,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &gt;1E4 
               
               
                   
                   
                 AN 
                 200 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall III 
                 390 
                 SA 
                 170,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 5 
                 EC 
                 2,500 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Liquid 
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 50,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &gt;1E4 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall III 
                 195 
                 SA 
                 70,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 4 
                 EC 
                 1400 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Liquid 
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 41,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 40 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germall III 
                 390 
                 SA 
                 76,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 IPBC 
                 8 
                 EC 
                 3,400 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Liquid 
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 14,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Unpreserved 
                 0 
                 SA 
                 &gt;1E6 
                 54,000 
                 4,400 
                 20 
                 &gt;1E4 
               
               
                   
                   
                 EC 
                 80,000 
                 67,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 PSA 
                 2,000 
                 4200 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 BC 
                 &gt;1E6 
                 &gt;1E6 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 CAN 
                 990,000 
                 320,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 AN 
                 380,000 
                 170,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 10 
               
             
             
               
                   
               
               
                 COMPARISON OF ACTIVITY OF GERMABEN II AND GERMABEN III 
               
               
                 (SCREENING EMULSION) 
               
             
          
           
               
                 Preservative 
                 Use Level 
                 Organism 
                 48 hrs 
                 7 days 
                 14 days 
                 21 days 
                 28 days 
               
               
                   
               
             
          
           
               
                 Germaben II 
                 0.30% 
                 SA 
                 480 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 20,000 
                 100 
                 2,600 
                 380,000 
                 380,000 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germaben II 
                 0.75% 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 100 
               
               
                 Germaben III 
                 0.30% 
                 SA 
                 7,000 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 14,000 
                 120 
                 &gt;1E4 
                 470,000 
                 190,000 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Germaben III 
                 0.75% 
                 SA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                 5 
                 EC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 PSA 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 BC 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 CAN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                   
                   
                 AN 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
                 &lt;10 
               
               
                 Unpreserved 
                 0 
                 SA 
                 &gt;1E6 
                 46,000 
                 &gt;1E4 
                 60 
                 &gt;1E4 
               
               
                   
                   
                 EC 
                 &gt;1E6 
                 170,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 PSA 
                 690 
                 24000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 BC 
                 &gt;1E6 
                 &gt;1E6 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 CAN 
                 440,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
                   
                 AN 
                 87,000 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
                 &gt;1E4 
               
               
                   
               
             
          
         
       
     
                                                                                         TABLE 11                   COMPARISON OF ACTIVITY OF GERMABEN IIE AND GERMABEN IIIE       (SCREENING EMULSION)            Preservative   Use Level   Organism   48 hrs   7 days   14 days   21 days   28 days                    Germaben IIE   0.30%   SA   580   &lt;10   &lt;10   &lt;10   &lt;10               EC   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               PSA   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               BC   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               CAN   1,600   &lt;10   &lt;10   &lt;10   &lt;10               AN   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10       Germaben IIE   0.75%   SA   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               EC   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               PSA   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               BC   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               CAN   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               AN   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10       Germaben IIIE   0.30%   SA   270   &lt;10   &lt;10   &lt;10   &lt;10               EC   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               PSA   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               BC   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               CAN   4,000   &lt;10   &lt;10   &lt;10   90               AN   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10       Germaben IIIE   0.75%   SA   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               EC   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               PSA   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               BC   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               CAN   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10               AN   &lt;10   &lt;10   &lt;10   &lt;10   &lt;10                    
Discussion of Challenge Testing Results
 
     The 28 day challenge results reported in Tables 3–11 above demonstrate the effectiveness of the preservative composition of the invention in a use emulsion composition against a wide range of bacteria and fungi organisms. 
     While the invention has been described with particular reference to certain embodiments thereof, it will be understood that changes and modifications may be made which are within the skill of the art. Accordingly, it is intended to be bound only by the following claims, in which: