\II M NH 1 1 iii. 7 A, ‘— \M ”01411121211111“ 3100? at)? THE 1311311311011 11511-101: ‘ AND ". t? «MENU; COEFFECHENT 11111.31: CF ‘E‘ES‘1‘ENG V1133: NAM DifiWFECTANTS 134313 for 1119 33-3130 011%??- $131011? ‘AN STATE 031.3... USE ; Anita “13111111 Leavzt’t I 1111; .I l . ,l g I I . I. . 1 c9 . . fij?‘[ 1". L (IC‘J u, . n .. . . , . . . . .u . . . . . . .. , . y .P . I. . .. .4 ..,. L... $13.31.... 1.! «I... ‘1‘. ....~o .. . . . . . . o . A 4 . ’ ‘_—~_—‘q—--—ru'. .1 d..- N . Lb}: ””1 at Ma“ 3’3 i w; ' (11;. ~. - “ \ . I . . I , . w ‘. -". .1 x ’ ' t ‘ -. - . ‘15:; 5 _I 1“a I. n. ‘ L‘I ‘ _ I . . L‘L 1-.I . “"352“ .. . I ‘ . - . k1”- ' 1!, “1"." ’ " . ‘ A . ~ 1 1’1“" l 1. . ' ‘ .. - p ‘ '7‘ . 51‘1"! ‘ r~ ‘u . l ' ll”? 1 .*L‘\ I, i ' ,~ '1') i .I '1‘ I I ~ I » ‘, ‘1 ‘, "1‘“ ‘ - g . " "1 1“ ? l J H ' ‘1 h 1‘ r 7t. . ‘ f , . ‘_. fa \' '. . 1 . 4 N " ifi ’7 y,» “M " 1" QA .3 “"3," ‘s t ‘p 1"?“ ‘JIZ‘ -. ‘ J, "I if -{ 1 'W ' . ’ " 3.9-. i‘ It... 9‘. p» N ‘4. I -__ _ _. -_ ; a ___ p .u‘,’ .1‘ 39‘s -. '6' I “x .p ., rJ 32-: _ 'w H ‘ . 'u‘T'L -,= .1 g‘ » a ' = '5‘ __-.-.'~ ur This is to certify that the thesis entitled "Comparative etudy of the Use-Dilution Method and the F. D. A. Phenol Coefficient Method of Kesting Veterinary Disinfectants". presented by Anita harriet Leavitt has been accepted towards fulfillment of the requirements for Major professor Date—AngnBLJM 9.454... GNPABATIVE STUD! 01’ THE USE-DILUTION METHOD AND THE F. D. A. PHEOL COEFFICIENT IIETHOD 0F TESTING VETERINARY DISINFECTANTS by ANITA HARRIET Lynn A WIS Submitted to the School of Graduate Studies of Michigan State College of Agriculture and Applied Science in partial fulfillment of the requirements for the degree of IASTER OF SOIIHCE Department of Bacteriology and Public Health 1946 . l I l ‘1‘I.L[.[[|II|J\'IIII [ {[[llll'llil‘r'llil II I ACKNOWLEDGE! The author viehee to acknowledge her sincere appreciation to Dr. I. L. lallnann for his interest and assistance in these studies. : "1 Fa Q J (I) d W: i. Il'l'RONGTIO Since it was first noticed that various compounds had an in- hibitory effect on the life cycle of some microorganisms, many methods have been tried to find a satisfactory means of neaeuring this inhibitory effect. As early as 1750 Pringle (1) carried out some experiments com- paring the value of different substances to hinder mtrefaction. Robert Koch in 1881 (2), however, was the first to compare the germi- cidal value of various disinfectants upon pure cultures of bacteria. He worked with emulsions of anthrax spores dried upon silk threads and estimated the time of survival in solutions of mercuric chloride. In 1889-91, Geppert (3 and I.) criticized Koch's thread method in view of the unavoidable carrying over of traces of disinfectant into the test culture; these traces were able to prevent growth of the organim adherent to the thread, although they night not have lost their viability at the moment of subculture. Grnber (5) in 1891 ad- vocated successive dilution of the test samples to overcome the nie- leading results from the carrying over of the mercuric chloride. In 1897 Kronig and Paul (6) devised a method of estimating the germicidal value of disinfectants, using garnets coated with anthrax spores as test objects. Kronig and Paul were the first to realise that the relative value of disinfectants depends very largely upon the conditions under which they are tested; they stated that in the comparison of disinfectants close regard should be paid to the “con- stancy of umber and species of bacterium used, constancy of taupera- ture, constancy of nutrient medium for test cultures, and absence of organic matter during disinfection" . In 1903 Rideal and Walker (7) proposed the "drop method" of testing disinfectants. This method measured not the absolute ef- fect of the disinfectant but its relative effect as compared with phenol. Anderson and Icclintic (8) in 1911 proposed a phenol coeffi- cient test known as the Hygienic Laboratory lethod. A number of changes were made in this method by the Conittee on Standard lethods of Examining Disinfectants of the American Public Health Association (9) in 1918; it proved to be too complicated and too expensive a method to be used routinely and was later abandoned by the Go-ittee. One important suggestion was made, however, namely that future work on disinfectants should include the use of disease producing types of bacteria other than m 3%. After ten years experience with both the Rideal-lalker and the Hygienic Laboratory methods, a modification was devised in the De— partment of Agriculture in which the best features of both were“ in- corporated and the most objectionable features eliminated (10). Shippen devised this method which, with few changes, was published later by Reddish, his successor. This procedure, known as the I. D. A. method (11) is most comonly used today in testing disin- fectante. It is hard to evaluate disinfectants on a comparative basis as we have no single reliable method of comparative testing. The F. D. A. phenol coefficient method indubitably gives couparable re- sults when testing coal tar products, but it cannot be used to compare the disinfectant value of a quaternary ammonium compound, a mercurial and a chloro—phenol. Tilley (12) has stated that '... a phenol coefficient has no practical value, whatever, except as a means of preventing the use of positively worthless preparations and as an aid to the manufacturer in maintaining the uniformity of his product”. Varley and Reddish (13) have stated that, “the phenol coefficient figure is a suitable measure for the practical value of phenol-like disinfectants when the factor 'twenty times the phenol coefficient' is aployed for calculating the dilution for use in practice". However, since the manufacturer usually recom- mends a dilution for his product, this recommended dilution should be used in evaluating the disinfectant. Also, this dilution should be based on definite test against the organism it is desired to kill. It is unfair to classify a disinfectant as poor merely be- cause its F. D. A. phenol coefficient value is low. At the recom-- mended dilution it may readily kill the desired organism in a time comparable to actual use. If it is not effective at this dilution a new use-dilution should be recomended. Hymn (11.) has demonstrated that the dilution coefficient should be taken into consideration when evaluating a disinfectant by its phenol coefficient. Upon diluting a disinfectant to one-half its original concentration, the disinfecting power may be reduced anywhere from two to sixty-four times depending upon the disinfectant. This factor of dilution could be eliminated if the disinfectant is evalua- ted at the manufacturer' s recommended use-dilution. The use-dilution method of testing disinfectants was devised in an effort to improve upon the F. D. A. phenol coefficient method and to evaluate disinfectants under conditions more comparable to their actual use. In disinfection the obj act to be treated is usually covered with layers of dried organisms. The glass rod used in the use-dilution technic is also covered with layers of the dried culture and, therefore, would more nearly simulate ac- tual use conditions for the disinfectant. The glass rods are rinsed before they are put into the culture medium so there should be less carrying over of disinfectant such as there is in the phenol coefficient method of testing. A comparative study of the 1". D. A. phenol coefficient method and the use-dilution method was undertaken. Veterinary disinfec- tants were used for two reasons; one, because most of them have a relatively low phenol coefficient which means that they have a smaller dilution range in which to work efficiently; secondly, veterinarians depend a great deal on disinfectants, and it was de- sired to know if these capounds were effective at the race-ended dilution. A few quaternary ammonium coupounds also were tested for comparison. W The test organisms used were a standard strain of W m and Wm strain 209, obtained from the Food and Drug Administration. AM was grown in standard broth described in 1'. D. A. Bulletin 198 (11) and m M. was grown in Difco disinfec- tant testing medium. n1. broth cultures were transferred daily and fresh transfers were made monthly from the stock agar slant culture. The M; m cultures, when grown in Difco disinfec- tant testing broth are always adjusted with llaOH to a pH of 7.0 to eliminate changing the pH of the disinfectant solution in the usual phenol coefficient procedure. Although, in the use-dilution pro- cedure, the culture is dried and would introduce no material change in pH when dropped into the disinfectant dilution, the culture was neutralized in order to keep it comparable in the two methods. Phenol coefficient tests according to the F. D. A. regulations were run on the disinfectants under study, using both Ea mag and m m as test organisms. In the use-dilution method, 22 to 26-hour broth cultures of Mm and gm are shaken for fifteen minutes to break up bacterial claps. Sterile glass rods, one inch in length and one-fourth inch in diameter, having a loop at one end for hand- ling, are dipped into the broth cultures and then carefully laid for drying on sterile filter paper in Petri dishes, care being ex- ercised so that the rods do not roll while drying. Iedication tubes contuning ten m1. of the test disinfectant and the tubes containing the rinse physiological saline are placed in a 20° 0., water bath. The bath is held at this temperature throughout the test. At the end of the drying period, four rods are dropped simul- taneously into each medication tube containing the test disinfectant. At intervals of one, five, ten and thirty minutes, a rod is removed and immersed for one minute in ten ml. of sterile physiological sa- line. It is then placed in ten ml. of nutrient broth. The tube is shaken vigorously to remove all organisms adhering to the rod and suitable dilutions are plated to measure quantitatively the extent of kill. Both the tubes and plates are incubated at 37° C. for 1.8 hours. Lack of growth at the end of this period has been accepted, in this thesis, as evidence of sterilisation. Controls are run by dropping rods covered with the dried or- ganisms into the rinse tubes of physiological saline for one minute and then into the tubes containing ten ml. of broth. Dilutions of 1:100 and 131000 are plated. The count is recorded as the total umber of organisms washed of the rod in the nutrient broth. The use-dilution is considered as the lowest concentration of a disinfectant that will kill in ten minutes but not in five. Hanes (15) stated that the drying time of the bacterial broth culture on the glass rods was not significant. After obtaining some negative controls when L m was used, it was decided to carry out some experiments involving this time factor. Inoculated rods were dried for varying lengths of time, then dropped in 10 ml. of physiological saline for one minute and then into 10 ml . of nutrient broth . Suitable dilutions were plated. The results are shown in Table 1. It would appear that thirty minutes is sufficient for dry- ing the rods inoculated with M2. am. A much lower count is encountered when L m is used. As no growth occurred after ninety minutes of drying and a fair count was obtained at twenty minutes, it was decided to use the shorter drying period for §._ m. Com atve c more; Although the use of phenol as a disinfectant has been general- 1y discontinued because of its low killing power and its toxicity, it has been maintained as a standard for the comparison of disin- fectants. The use-dilution against §_t_apl_i_,_ m was betwoen 1:70 and 1:80 as compared with a dilution of 1:60 using the F. D. A. phenol coefficient method (Table 2). It is interesting to note that M m was killed in less than one minute at a dilution of 1:60. An examination of Table 3 shows that am was killed at 1:110 by the use-dilution method whereas by the phenol coefficient method it was 1:90. lam was killed at 1:80 in one minute. The use-dilution required a greater concentration of phenol for kill than the F. D. A. method, but the difference in concentration is not very great. m1. Amphyl, which is made up of chloro-xylenol, butyl and amyl phenol isomers and soap in a 20 per cent alcohol, is recommended by the manufacturers to be used in a dilution of approximately 1:160. Table 5 shows thatt. this dilution kill was not obtained in ten minu- tes against m m. The use—dilution was 1:100 as compared with 1:250 by the phenol coefficient method as shown in Table 6. Tables 4 and 5 show the variation that was obtained in the control counts. Although the control gave a count of 850,000 in one test and only 40,000 in the other, the culture tubes from the 1:100 di- lution of disinfectant gave the same result in both tests. An evaluation of Tables ’7 and 8 shows a use-dilution of more than 1:700 and less than 1:800 as compared to a phenol coefficient dilution of 1:850 against §_._ m. The manufacturer's race-en- ded dilution gave mediate kill . Amines Aseptogen which is ortho phenyl phenol, iso propyl alcohol and soap is reccnended by the manufacturer to be used in a dilution of 1:20. lhen tested with em 521.1122. the use-dilution was less than 1:5 (Table 9) as compared to a phenol coefficient dilution of 1:10 (Table 10). The manufacturer's recommended dilution of 1:20 failed to kill even after 30 minutes exposure. When tested with E; m aseptogen had a use-dilution of more than 1:200 and less than 1:300 (Table 11) and a phenol coefficient dilution of 1:150 (Table 12). Phenolor Phenolor which is also a flienol derivative is recomended for use at a dilution of 1:30. Although this dilution did not kill m gum in one minute, it did in five minutes. Examination of Table 13 shows that the use-dilution was 1:100 against _S_t_am,_ 52% while Table 1!. show that the phenol coefficient dilution was 1:150. Using E; m as the test organism the results are shown in Tables 15 and 16. The use-dilution was 1:400 and the phenol coefficient dilution was 1:300. In general, the concentration of these disinfectants required to kill m m in ten minutes was greater using the use-dilution technic than it was by the phenol coefficient nethod. However, when £31.19;th was the test organis- the results tend to show that the use-dilution was less concentrated than the phenol coefficient dilu- tion. This may have been due to the fact that Q, ms; was not as resistant to drying as flag; m, and it was difficult to get comparable results, even in the controls. kgol and Qtigeptine Both lysol and antiseptine are cresylic acid preparations. Lysol is generally used in a 1:100 dilution while the recommended dilution for antiseptine is 1:40 . Examination of Table 17 shows that lysol was effective in five minutes at its race-ended dilution. Its use- dilution against M m was 1:200 as compared with its phenol coefficient dilution of more than 1:150 and less than 1:200 (Table 18). As shown in Tables 19 and 20 both the use-dilution and the phenol co- efficient dilution of lysol were 1:600 against L W. Results on testing antiseptine against _S_t_a_pl_1_._ m are shown in Tables 21 and 22. M, m was not killed in five minutes 10 at the remanded dilution, but it was in ten. Its use-dilution was 1:50 and its phenol coefficient dilution was 1:60. Using A m as the test organism the use-dilution was 1:400 (Table 23) and the phenol coefficient dilution was 1:350 (Table 24) . lith both of these disinfectants, comparative values were obtained by the two methods of testing. lein Creolin, a coal tar product, is remanded to be used at a concentration of approximately 1:80 . As shown by Table 25 growth of m m occurred even after ten minutes exposure to this concentration; a lower dilution which would produce bacterial kill in a shorter length of tine would be more satisfactory for general use. The use-dilution against 33.99.: §_u_r_e_u_s_ was 1:40 and the phenol coefficient dilution was between 1: 50 and 1:75 (Table 26) . Examina- tion of Tables 27 and 28 show that using L m as the test or- ganisn the use—dilution was 1:900 and the phenol coefficient dilu- tion was between 1:800 and 1:900. Dr. Hess Dip Dr. Hess Dip is also a coal tar derivative. It is evident from the results obtained in Table 29 that the recommended dilution of 1:70 would exhibit no germicidal effect against £13.34. 993311; after thirty minutes exposure. Its use-dilution was 1:25 as compared to a phenol coefficient dilution of 1:60 (Table 30). Results from 11 Tables 31 and 32 show that both the use—dilution and the phenol co— efficient dilution was 1:400 when L M was used for the test organism. W Liquor cresolis saponatus, also a coal tar product, was effec- tive against £13.92; m at its reed-ended dilution of 1: 50 after one minute exposure (Table 33). Its use-dilution was more than 1:50 and less than 1:75 against M M while the phenol coeffici- ent dilution was 1:80 (Table 34). Tables 35 and 36 show the results obtained using L Me; as the test organism. The use-dilution was 1:200 as compared to a dilution of 1:150 using the phenol coef- ficient technic. An evaluation of these results again show that the use-dilution was lower than the phenol coeffici ent dilution when Staph. M was used but that it was higher when A m was the test organism. ' zgngleum Zenoleun, the composition of which is not stated, is recommended for use at a dilution of 1: 50. Examination of Table 37 shows that a dilution of 1:20 was needed to effect kill in ten ninutes against m m by the use-dilution technic while a dilution of more than 1:20 and less than 1:30 was required using the phenol coefficient method (Table 38) . When L 3129—03: was used the use—dilution was be- tween 1:300 and 1:400 (Table 39) and the phenol coefficient dilution was 1:350 (Table 40). 12 An examination of Table 41 shows the results obtained on all the forementioned phenol-like compounds. The results are fairly comparable for the use-dilution method and the F. D. A. phenol co— efficient method. In general, a slightly lower use-dilution was obtained when m m was used. When g; m was the test organism, slightly higher dilutions were obtained for the use- dilution than the phenol coefficient dilution. It is noted that of the ten disinfectants studied in this group, only five have a recom- mended dilution that is below the use-dilution. Phenolor was the only compound that had a remanded dilution that was five times its phenol coefficient dilution when tested against M em. Five times the phenol coefficient dilution is recognised as being that dilution which falls within the range of effective disinfection. When E; M was the test organism, the majority of these disinfectants had a race-ended dilution with- in this range. However, in general disinfection, it is desirable to kill gram-positive organisms of which mg m is representa— tive, as well as gran—negative ones, represented by h m. Exa- mination of Table 42 shows that all the disinfectants except senolem killed E: me. during one minute exposure. Zenoleum was effective within five minutes. If disinfection was merely against gran-nega- tive rods they all could be rated as efficient. On the other hand, three of them, eseptogen, Dr. Hess Dip and senolenn did not kill Maggie even after thirty minutes exposure. Alphyl and creolin killed m manor ten minutes which is too long for general use. Antiseptine was effective after five minutes but still does not 13 have a recommended dilution that is five times its phenol coeffi- cient dilution against M, w. Phenolor, lysol and liquor cresolis saponatus were effective after one minute exposure; phenol was the only one that killed £3.29; m within one minute. 14 Benchlogen and Dowicide C Dowicide C which is chloro ortho phenyl phenate and Benchlophen which is a mixture of monobenzylated ortho chloro phenols and mono- bensylated phenols provided some interesting results when tested with M m. The use-dilution for- Benchlophen was 1: 50 (Table 43) while by the phenol coefficient method the killing dilu- tion was 1:1300 (Table 44) . lhen _E;_,_ m was the test organism the results were more comparable, as shown in Tables 45 and ‘46, the use-dilution being between 1:150 and 1:200 while the phenol coef- ficient was 1:200. Dowicide 0 gave similar results in that its use-dilution and phenol coefficient were both 1:5000 against E; m (Tables 49 and 50) Idlile its use-dilution using £221.; m was 1:100 (Table 1.7) while its phenol coefficient dilution was 1:5000 (Table 48). Although Dowicide C in this form is not intended to be used as a disinfectant, it was included because it was desired to sum a cupound having a phenol coefficient between the phenol—like com-- pounds and the quaternary amnion preparations . 15 Ammonium Con d Quaternary ammonium compounds are known to give "spotty" re- sults when tested by phenol coefficient methods. In order to as- certain whether more uniform results could be obtained by the use- dilution technic, tests were run on B. T. 0., tetrosan and emulsept. By examining Tables 51, 55 and 59, it is evident there was enough carry over of the disinfectant into the test medium to prevent growth of M; w in the broth tubes. Growth did occur on the plates. That may have been due to the fact that dilution was made beyond the point of bacteriostatic action. Another theory is that the positively charged quaternary ammonium compounds hook onto the negatively charged bacteria; on vigorously shaking the tube of broth, enough of the disinfectant might be shaken off to allow some of the organisms to grow if they were plated immediately. All of the tubes that showed no growth at the end of 48 hours incubation, when tetrosan was tested with M 91339;, were inoculated with m m and again showed no growth at the end of 48 hours in- cubation, indicating bacteriostatic action. There was an indication that lower dilutions would be obtained when the use-dilution technic was used than when the phenol coefficient method was mployed. llore work needs to be done before definite conclusions can be drawn. M It is evident from the concluded experiments that most manu- facturers base the recommended dilution on its killing power against L, m. for general disinfection this is not enough. It should be based on tests against the more resistant gram—positive organises which are apt to be encountered. M; w is rep- resentative of this group.. In order to have a.margin of safety the recommended dilution should be five times the phenol coeffi- cient dilution or the use-dilution for.§§gpgguggz§n§e lhen phenol-like compounds were tested by the use-dilution method, slightly lower dilutions usually were obtained than by the filenol coefficient technic when m m was used. lhen E; jyphg§9,was used, as a rule slightly higher dilutions were obtained by the use-dilution method of testing. Benchlophen and Dowicide 0, both chloro phenols gave similar results for the two methods when £5.312hosa was the test organism. lhen m m was used very much lower dilutions were obtained by the use-dilution technic than.by the phenol coefficient technic. Indications are that lower dilutions would be obtained when testing quaternary ammonium compounds by the use-dilution.method than by the phenol coefficient method, but more work needs to be done on these compounds. W l. Pringle: Some Experiments on Substances Resisting Putrefaction. Phil-e We, vale XLVI, me 2. Koch, Robert: Ueber Disinfection. Mittheil, A. I. kaiserl. Gesundheitsampte, vol. I. 3. and 4. Geppert: Zur Lehre von den Antisepticis. Berl. klin. Wachenschr, 789 and 819, 1889. 5. Gruber: Transaction of the VII Internet. Congress of Hygiene and Demography, Vol. II, pp. 255. Centralb. of Bakt. l, Abt. Vol. II, 115, 1892. 6. Kronig, B. , and Paul, T. L.: Die chemischen Grundlagen der Lehre von der Giftwirkung und Disinfection. Zeitechr. f. Hygiene, Vol. m, 1. 7. Rideal, S. and Walker, J. T. A.: The Standardization of Disin- fectants, Jour. Roy. Sanitary Inst., 24, 424, 1903. ' 8. Anderson, J. 1". and llcClintic, T. B.: A lethod for the Bacterio- logical Standardisation of Disinfectants. J our. of Inf. Diseases, 8, 1, 1911. 9. Reddish, George: Determining the Germicidal Efficiency of Dis- infectants. A. J'. P. H. 16, 283, 1926. 10. Reddish, George: Examination of Disinfectants. A. J. P. H. 17, 320, 1927. 11. Ruehle, G. L. A. and Brewer, C. IL: U. 8. Food and Drug Admin- istration Hethod of Testing Antiseptics and Disinfectants. U. 8. Dept. of Agriculture Circular #198, 1931. 12. Tilley, F. IL: Phenol Coefficients. A. J. P. H. 11, 513, 1921. 13. Varle and Reddish: Disinfection and Sterilization, 2nd edition (llcCulloch), page 186. 14. Hymn, Andrew H.: To Devise a Method of Expressing the Bacteri- cidal Efficiency of a Disinfectant Other than the Standard 1'. D. A. Phenol Coefficient Method. II. S. Thesis, Hichigan State College, 1940. 15. Hence, 1!. E.: The Use-Dilution Method of Testing Disinfectants. I. S. Thesis, Michigan State College, 1945. Resistance of L m to drying Drying period tube Plate counts 20 minutes + 14,000 12,000 30 minutes + 6,000 7,000 60 minutes + 19,000 17,000 90 minutes - 0 0 120 minutes - 0 0 Resistance of Stem. aureus to drying Drying period tube Plate counts 30 minutes + 1,130,000 1,190,000 60 minutes 1» 1,120,000 1,0m,000 90 minutes + 1,300,000 1,430,000 120 minutes 4» 1,480,000 1,550,000 150 minutes + 1,090,000 1,330,000 Test compound - Phenol Tut crowin- - sum. m *Use dilution - > 1:70 (1:80 Dilution 1-60 1-60 1.70 1.70 1-80 1.30 1-90 1-90 Control Counts Time of Exposure ma 1 minute 5 mimlte_s__ 10 minutes m 539,,“ B gut; Broth 3.91;;— s 10:? 0 “thump. *‘T‘ o - 0 0 — 0 - 0 - 0 260 - 0 - 0 - 0 290 - 0 - o - 1 o 5030 + 10 + O - O 4520 + 10 + 0‘ - O In. + 30 + 10 - 0 In. + ( so + 0 - 0 620,000 810,000 me P. D. A. phenol coefficient method at a dilution 1:60 effects kill in ten minutes but not in five. of Test compound - Phenol Test organism - §_._ m ‘lUse dilution - 1:110 Time of Exposure 1 minute Lminutes 10 minutes 30 minutes Wu Plate Plate Plate ‘ Dilution m M ro C t 'MW m 1.80 - 0 - 0 - O -- 0 1-80 - o - 0 - o - 0 1-100 + 0 - 0 - 0 - 0 1-100 + 0 - 0 - O - 0 1-110 + 130 + 0 - 0 - 0 1-110 + Z!) + 0 - 0 - 0 1-120 + 210 +- 30 + O — 0 1-120 + 500 + 20 + 0 - 0 Control 54,000 # , Counts 141,000 *The P. D. A. phenol coefficient at a dilution of 1:90 effects kill in ten minutes but not in five. 3.. .- : arsflflr|lsilbllflli 0‘ . a. . . ,..I»II Hf . Test compound - Amphyl Test organism - taph. m Use dilution - 1:100 Ianufacturer' s recommended dilution - 1:160 Tine of Exposure . 1 minute tes 10 minutes Plate Plate Plate Plate Dilution 1 coun 8mg} Count th 0 t Bro 00 t 1-50 + 230 - 0 - 0 - 0 1-50 + 240 - 0 - 0 - 0 1-75 + 330 - 0 - 0 - 0 1-75 + 270 - 0 - 0 - 0 1-100 + 220 + 20 - 0 —‘ 0 1-100 + 410 + 10 - 0 - 0 Control 850,000 Counts 600,000 ' Test compound - Amphyl Toot cranni- - £22111 same Use dilution - 1:100 Dilution 1-100 1—100 1-160 1-160 1-200 1-200 1-300 1-300 Control Counts Time of Exposure l unute 5 minutes 10 minutes 30 minutes a an m1 3.1;: 14:13,: m. PM: Lgrfi 500 «I» 150 - 1 0 - 0 + ' 600 + 190 - o - 0 + 900 l + 160 + 0 - 0 +- 980 + 1 150 + O - O T 3000 + 150 + 10 - 0 .. 3500 + 190 + 20 - 0 4. 3600 + 240 + 10 - 0 + 8500 + 260 4- 20 - 0 1+0 ,000 42,000 Test compound - Amphyl Test crania- - flame am Phenol coefficient dilution - 1:250 Time of Exposure Dilution of M1 5 minutes 10 minutes 15 minutes 1-50 - - '- 1-60 + - - 1-70 + + .. Dilution of Anphyl ' 1-100 - - - 1-200 - - - 1—250 1» - - 1-300 + + .. 1-350 + 6 + + 14.00 + + 4- 1-450 + + «- Test coupound - Alphyl Test organism - Eberthella mg Use dilution - 71:700 <1:800 Time of Exposure ,rhulmla. M Dunn» wig. Bro rent“? :3 fiflmfiifil 1—160 - 0 - 0 0 - 0 1-160 — 0 - _ 0 o - 0 1—700 4- 110 - 0 0 .. 0 1-700 4» 160 - 0 0 - 0 1-800 + 6100 + 10 0+ - 0 1-800 , 4- 5600 + 20 0 - 0 1-900 + L Innj 1+ so 0 - I 0 1.900 + In. + 110 10 _ - r 0 Control 568,000 _ Counts 37,000 V fill-L3. Test compound - Amphyl Test organism - Eberthella M Phenol coefficient dilution - 1:850 Dilution Phenol 1-80 1-90 1.100 of Amphyl 1-700 1-750 1-800 1-850 1-900 1-950 1-1000 Time of Exposure ‘ 5. mimic; 10 mt» 15 mg; , up -. e. + + - + C c- 4- + - — d- 4- + + + + nun: Test compound - Aseptogen Test organism - Stag. m Use dilution - < 1:5 lanufacturer' s recommended dilution - 1:20 Dilution 1-5 1-5 1-10 1-10 1-20 1-20 1-40 1-40 Control Counts Time of Exposure 1 minute 5 minutes 10 W W Plate Plate Plate Plate 0 t B th Count Broth, Count MM + 600 + 70 + 0 - 0 + 2800 + 60 + 10 - 0 + 1430 + 130 + 1+0 - 0 1 + 3710 + 150 + 10 - 0 + 5520 + w1690 + 120 + 0 + 4500 + 1260 + 100 + 20 +- 7500 + 1630 + 630 + 90 + 8000 + 1960 4- | 500 + 70 487,000 439'“ I F l Test compound - Aseptogen Test organism - tag. auregg Phenol coefficient dilution - 1:10 Time of Exposure Dilution of Phenol . minutes 10 minutes 15 Ligteg 1-50 .- - 1-60 - .. 1-70 + + Dilution of Aseptogen 1-5 - .. 1-10 «I- - 1-20 1- +- 1-40 + 4- 1-80 4- + 1-100 4» 4- 1-200 + + Test compound - Aseptogen Test organism - Eberthfl m Use dilution - >1:200 (1:300 Time of Emosure _ 1 minute 5 minutes 1L minutes 30 minutes Dilution B £324 th £13111, Broth :1“: 33191.33 ' 1-50 ‘ Fijg o FEE-1 o 77% - 0 1-50 - 0 -— 0 - 0 - 0 1-200 + 90 - 0 - 0 - 0 __ 1200 + 220 - 0 - 0 - 0 1-300 + 5770 + 0 + 0 - 0 1-300 + 4250 + ' 0 + 0 - 0 14.00 + 8100 + 790 + 0 - 0 14.00 t + 7720 4- 510 + 0 1 - 0 Control 20,000 7 L Counts 24,000 W Test compound - Aseptogen Test organism - Eberthella M Phenol coefficient dilution - 1:150 Time of kposure Dilution of Phenol 1 5 li___m;tefl 10 minute 15 giggteg 1-80 - .. 1 1-90 + 4- 1.100 + +- Dilution of Aseptogen 1-50 - .. 1-100 - .. 14.50 + .. 1-200 + 1- 1-250 + + 1-300 + 4- 1-350 + + 1-400 + + Test cmpound - Phenolor Test organism - m, aureus Use dilution - 1-100 Ianufacturer' s recommended dilution - 1-30 Time of Exposure 1 to 5 5%“: 10 tes 0 mute Bro Plat: Broth 10013:: BrothEi B oth P0681“ D1136” J + 0 — i 0 L._._:..1 0 hit-1% 1.30 + 0 - 0 - 0 - 0 1-50 + 0 - 0 — 0 - 0 1-50 + 0 - w—0 - 0 - 0 1.100 + 130 + 10 - 0 - 0 1100 + 150 + 20 - 0 v — 0 1.200 + 2,000 + 100 + 0 - 0 1-200 + 7,000 + '70 + 10 - 0 Control 780,000 Count 670,000 Test canpound - Phenolor Test organism - .1922; 9122112 Phenol coefficient dilution - 1:150 Time of hposure Dilution of Phenol . 2 plates 10 minutgs 15 mt» | 1-50 - i - .. 1-60 +——~ .. _. 1-70 + + 4- Dilution of Phenolor 1.25 - c- .- 1-100 ‘ - - .. 1-150 + .- .. 1-200 + 1. .. Test compound - Phenolor Test organism - Eberthfl Mpg; Use dilution - 1:400 Dilution 1-200 1-200 1-300 1-300 1-400 14.00 1-500 1-500 1-600 1-600 Control Counts renal: Time of hposure 1 minute 5 minutes 10 minutes 30 minutes Plate Plate Plate Plate .. 0 - 0 - 0 - O o- 0 - 0 - 0 - O + 0 - 0 - 0 - o + 530 t- 0 - 0 - o . 1.30 + 9 - o - o L + 850 + 0 + 0 - 0 + 760 + O + 0 -- 0 k 4» 2450 + 90 + 0 + 0 + 191.0 + 70 + 10 + 0 20,000 20,000 llllrr£EEJB ll. 1.. bu v? n ‘ Test cmpound - Phenolor Test organim - flagella M Phenol coefficient dilution - 1:300 Time of Exposure Dilution of Phenol 5 minutes 10 minutes 15 minutes 1-80 + - - 1-90 + v " 1-100 4- + - Dilution of Phenolor 1-100 - - - 1—200 - - - 1-300 + - - 1-400 + + '- 1.5“) + + "‘ 1-600 + + 4- Test compound - Lysol Test organism - tag. aureus Use dilution - 1-200 lanufacturer‘ s recouended dilution - 1-100 Time of Exposure 1 minute minutes 19 mu; 0 minutes Plate Plate Plate fim Dilution ,8 t mmmamiaamama 1-50 - 0 - O - 0 - 0 1-50 - O - O - 0 - 0 1-100 + 0 - O - 0 - 0 1-100 + 0 - O - 0 - 0 1-200 4» 960 + 10 - 0 - 0 1-200 4- 420 1- 10 - O - 0 1-300 + 3730 4- 50 + 0 - 0 1-300 + 4450 f- 30 + 0 - 0 Control 740,000 Counts 520,000 TAPE—1.8. Test compound - Lysol Test organism - fl, gureus Phenol coefficient dilution - >1:150 (1:200 Time of Exposure Dilution of Phenol 1 minute 5 minutes 10 minutes 1-50 - .. .. 1-60 + - .fi 1-70 F + . . Dilution of Lysol 1—50 - - f 1-75 - - - 1-100 - - .. 1-150 - .. _. 1.200 + .. _ ._ 1-300 + + 4- 14m + + + Test compound - Lysol Test organism - Mel}; twee Use Dilution - 1-600 Time of Exposure ‘ ; giants m to 1 ts W Plate Plate Plate Plate Dilution , Broth Count Broth l Cgunt ro Co B C t 1-500 + 30. - 0 - 0 - 0 1-500 + 10 - 0 - 0 - 0 1-600 + 980. 1. 30 - 0 - 0 1-600 4. 780 + 50 - 0 - 0 1-700 + 2760 + 10 + O - 0 1-700 + 3500 1- 10 + 0 - 0 Control 35,“ 00131“ 39,000 11211.22 Test compound - Lysol Te at cream!- - W M Phenol coefficient dilution-1:600 Time of Exposure Dilution of Phenol 5 minutes 10 minutes 15 minutes 1-80 - d - - 1-90 + - W - 1'10° 1- 4- .. Dilution of Lysol 1-400 - - - 1.150 .. .. .. 1-500 + - - 1-550 + r '- 1—600 4- - - 1-650 + + + 1&2}. Test compound - Antiseptine Test organism - £13211: M Use dilution - 1:50 Manufacturer's recommended dilution - 1:40 Time of Exposure 1 minute J minutes 10 minu 0 minute Plate W fl Plate Dilution MM Broth Cou_nt__ Broth Count M ML 1-25 + 10 - 0 - 0 - 0 1-25 + 60 - 0 r - 0 - 0 1-40 4- 200 + 0 - 0 - 0 1-40 4- 170 1- 0 - 0 - 0 1-50 4- 330 + 20 - 0 - 0 1-50 4- 100 + 10 - 0 — 0 1—100 4- 1610 f- 80 + 10 - 0 1-100 4. 1200 4- 50 4- 10 - 0 1-200 4- 15 .000 1- 4980 + 750 + 30 1.200 4 9000 4- 31.00 + 400 +* 70 Control 562,000 Counts ‘ 6007000 ‘— £311.22 Test compound - Antiseptine Test organism - __§§ph_,_ aurer Phenol coefficient dilution -- 1:60 Time of moms Dilution of Phenol ‘ 5 minutes __ 10 mi_m_lt_e_g 15 minutes i 1-50 - - 9 .. 1-60 - - 1 - 1-70 + + .. Dilution of Antiseptine 1‘20 - - - 1‘30 - - .. 1—40 - - - 1-50 + - -- 1-60 + -. .. 1-70 + ‘ + .. 1-80 + + ‘— _. 1.100 .. ' ' 1. 1 Test compound -.Antiseptine Test organism - Eggrthglla‘typhggg Use Dilution - 1-400 Dilution 1-300 1—300 1<400 1-400 1-500 1-500 Control Counts Time of Exposure 1 minute 5 minnggs [_Jfllgg%ggtggW Plate Plate Plate Plate igggigg,0032t. ngfth#929:§_ .Broth'Coznt Brothrgggn§_ + 10 - 0 - o - o 130 ex 0 - O — 0 100 + 0 - O - 0 190 + 10 4;: 0 - O } 210 + 10 + 0 - 0 i 50,000 45,000 Test coapound - Antiseptine Test organism - 59erthella typhogg Dilution of Phenol 1-80 Time of Exposure 15 g5ggtes 1-90 1—100 Dilution of Antiseptine 1-50 14100 1-200 1-250 1-300 1-350 19400 1-500 + Test calpound - Creolin 112L121 Test organism - mg gureg Use dilution - 1:40 flanufacturer' s reconended dilution - 1:80 Dilution 1-20 1-20 1-40 1-40 1-80 1-80 1-100 1-100 1-200 1-200 Control Counts Time of Exposure 1 minute minutes 1 t rflfimfi , 175 - 0 - 0 - ° + 330 + 0 - 0 - 0 + 2'75 1- 10 - 0 - 0 + 1500 +- 40 + 0 " 0 + 2100 + 40 + 0 - 0 + 7950 + 1150 4- 100 - 0 4* 4650 1- 780 + 90 - 0 + Inn. «r 8200 4- 6710 + 410 + Inn. «I» 7950 + ‘ 5100 + 1 610 340,000 890,000 M Test coupound - Creolin Test organism - Jame m Phenol coefficient dilution - 71: 50 < 1:75 Time of Exposure Dilution of Phenol .___.§____tt__1m1n tea 1121112219.; M. 1-50 - .. 1-60 4- - 1—70 + + 1-80 + 1. Dilution of Creolin 1-25 - .. 1—50 - .. 1.75 + 1. 1-100 + 4- 1-200 + 4- 1w 4' 1- Test compound - Creolin Test organism - Earthella we; Use dilution - 1:900 Dilution 1-800 1—800 1-900 1-900 1-1000 1-1000 Counts Time of Exposure 1 minute minutes 10 tes 29 m; Broth 10:32. Broth Egt‘ B;othi101a:;u Broth £313; .11 W, ‘71 o - o r“? o 1- 2830 - 0 - O - 0 + 4310 + 0 - 0 - 0 + 4600 + 0 - 0 i- 0 + In. + 1000 + 20 - 0 + In. + 1090 + 10 - 0 #580,000 \ 350,000 i Test compound - Creolin Test organism - Eberthella tmhosa Phenol coefficient dilution - Dilution Phenol 1-80 1-90 1—100 Dilution of Creolin 1-500 1-600 1—700 1—800 1-900 1-1000 >1:800 < 1:900 Time of Exposure 5 minutes 10 minutes 15 minutes .1 .. .. + + + t + ' + + +- + Test compound — Dr. Hess Dip Test organism - M gureus Use dilution - 1:25 Hanufacturer's recommended dilution - 1:70 Dilution 1-10 1-10 1-25 1-25 1-50 1-50 Cont‘rol Cougts Time of Exposure , 1 min to minutes 10 minutes 0 ‘utes I (Plate Plate Plate late Bgogh Coun} ngth ngnt ‘ B Count Bro C unt 1» 30 - 0 - 0 - 0 1- 50 - 0 - O - Oi + 210 + O - 0 - 0 + 520 + 10 - 0 - 0 + 1650 + 160 + 60 + 0 + 2510 + 160 + 30 + ’ 0 540,000 610,000 ‘ Teet compound - Dr. Bees Dip Test organism - m m Phenol coefficient dilution - 1:60 Time of Exposure Dilution of Phenol . 2 gauge 19 Mtge mi 1-50 - — .. 1-60 1- - - 1-70 + 1- Dilution of Dr. Bees Dip l-lO - .. 1.20 - W .. 1—40 - .. .. 1-60 + .. .. 1-80 + + + 1.100 4- 4. 1-200 ‘ + + +- Teet coupound - Dr. Hess Dip Test organis- - Emwelle 3% Use dilution - 121.00 Dilution 1-200 1-200 1-300 1-300 1-400 1-400 1-500 1-500 Control Counts Time of Exposure Plate Plate 1 Plate Plate 1.9.1292. Gong: Bro Count Broth C t Bro m .. 0 - O - O - 0 .. o .. o - o - o + 20 + 0 - 0 - 0 +- 10 + 0 - 0 - 0 + 60 + O - O - 0 + 50 + 0 - 0 - 0 + 650 + 0 + 0 - O + 820 1- O + 0 - 0 I 380,000 140,000 Teet conpound - Dr. Heee Dip Toot cranium - M1221 m Phenol coefficient dilution - 1:400 Tine of Exposure Dilution of Phenol 5 minutes 9.0 gm; ;5 m: 1-80 1 + - 1-90 + , .. 1-100 4- + Dilution of Dr . Hess Dip 1-2100 — ‘ .. 1-200 - .. 1-300 1- - 1-400 {- - 1-500 w + + I.“ L 1- 1- W Test compound - Liquor Creeolie Seponetue rut organism - m 3242922 209 Use dilution - ) 1-50 < 1-75 lumfecmrer' e recomended dilution -- 1-50 Dilution 1-25 1-25 1.50 1.50 1-75 1-75 1400 1-100 Counts 1 linnte ninute W minute: Plate Plate Plate Plate firothllgount roth Count Broth (3th ! ro Co - o .. o - o 4 O - If - '5 - o o + 30 - o - o f o + 60 - 0 .- 0 0 + 31.10 «I- 140 . + 0 0 I 2180 + 50 + 0 0 + In. + 700 + 180 0 4 In. + | 141.0 } T 120 o r 460,000 640,000 W Test compound - Liquor Cresolis Saponatus Test organism - §_,_ sureus Phenol coefficient dilution - 1:80 Time of Exposure Dilution of Phenol 5 minutes 10 minutes 15 minutes 1-60 - - - 1-70 1 1' + '- Dilntion — of Liquor Cresolis Seponet‘us 1-80 + .. - 1—90 + + - 1-100 + + 4. 1-110 + + + 1-120 «5- + 4- 1-130 + + 4. 1.140 - + + + W Test conponnd - Liquor Cresolis Seponetus Toot arsenic! - Mlle. 122192 Use Dilution - 1-200 Dilution 1-100 1—100 1-200 1-200 1-300 1-300 Control Counts ‘ minute Plate Time of Exposure tes 10 5%qu I i Plate Plate +2333; Cogt £39311 Count #1:; Count Bro let; .. o - 0 - 0 - 0 .. 0 - 0 - 0 - 0* + 6500 + 2750 - 0 - 0 + 5800 + 1250 - 0 - O + Inn. + In. + 850 - 0 4- In. 1 + Inn. + 1.20 _- 0 870,000 — W Test compound - Liquor Cresolis Seponstus Test organism - Eberthgga m Dilution of Phenol _5 Ignites 10 utes 1: mm , 1-80 - .. - 1-90 4- - .. 1-100 + t + + Dilution of Liquor Cresolis Seponatus 1-150 4» - - 1-200 + + + 1-250 4- 4» 4. 1'300 + + + 1-400 + + 4- 1-500 +- + 1- Time of Exposure Test conpound - Zenoleun Test organism - §tem. egreus Use dilution - 1-20 11010.21 lenufacturer' s recomended dilution - 1—50 Dilution 1-10 1-10 1-20 1-20 14.0 1-40 1-50 1-50 Control Counts Time of Exposure 1 minute minutes 10 Q grates Plate Plate iPlete { Plate Broth Cmt Broth Com , Bmth Count Broth Count 1 4. 300 + | 10 — 0 - 0 + 180 + 10 - 0 - 0 + 150 + 20 - 0 - 0 4» [.80 + 10 - O - O + 21.20 I + 80 + 10 - 0 + 2600 + 120 + 10 - O + 3500 + 360 + 30 + o + 1.200 + y 220 + 90 + 0 330,000 1 m,” m Test compound - Zenoleu Test omnis- - £229.. 99:93.9; 209 Phenol coefficient dilution - >l:20 (1:30 Tine of Exposure Dilution of Phenol 5 minutes _ 1_0__mim1tes 15 mimtes 1-50 - - . - 1—60 + + - 1‘70 + + 4 Dilution ”“ of Zenoleun full st. _ 1-5 -—- - - 1-20 - - - 1-30 + + + Test conpound - Zenoleun Test organism - Eberthelle typhose Use Dilution - >1—300 < 1-400 Tine of Exposure minutes tes Plate Plate OOOOOO Dilution M 993;: Broth 1-200 .. 0 1-200 4» 10 - 0* 1-300 + 1400 - 0 1-300 + 1; 60 - 0 1-400 + 5500 + 10 14.00 + 3990 + } 20 Control 200,000 290 , 000 ‘ Counts ' 700,000 ‘ 730,000 1 Test compound - Zenoleun Test organism - Eberthella moss #6 Phenol coefficient dilution - 1:350 Dilution of Phenol 1-80 1-90 1-100 Dilution of Zenoleun 1—100 1—200 1-250 1-300 1-350 1400 Time of Exposure 5mm low 15M...— 1. - a. + + +- 4. .. .. 4. — .. + + + omuav ONSA ‘ om: _ 8.3080? can; _ A on: _ §H30N . anaconda 2.3083“ 8: 8a; 033 on: rinses page «N: 83 SSH 8.3 2.: 03 30m .8 35.... mnuav out: 80“..” oomuVoowuflA own.” 3300.5 8m: STEVE»? 8o: 803 8”: H83 on: 83 . 8.3 can: 93 058832 83 8.3 8.3 80” on; Sausage 832.3“ 83 on: 83 on: Hosea «3 v 33 8n58~3A 033 on: nonsense 81H 8m£v8tfi one; 8.3 315 {i h . Home? _. .u 4. 8333 u . o .m . 33a 0% g 33 Seances l 033mg 0.33090 .8qu 35 «com, . .ha c.3095 H83 .flfiaufifi. hedge—E H223 someone: H585 3305803 o .8233.— 3.11.3.4: Test compound - Benchlophen Test organism - tag. aureus Use dilution - 1:50 Manufacturer' s recomended dilution - 1:200 Time of Exposure 1 minute 5 minutes 10 minutes 0 minute Plate 1513?;— Plate Plate Dilution roth Count 1% Count FEE-Egg 39.9.9.5... Broth 3931}; 1-25 + O - O - 0 - 0 1-25 + 10 - 0 - 0 —- ‘0 1-50 + 400 + 300 - 0 - 0 1-50 + 200 + 120 - 0 - 0 1-100 + 1050 + 345 + 80 -- 0 1.100 + A 1230 + 1.70 + 120 - 0 1—200 4» 2000 + 11.00 + 1.00 + 30 1-200 4» 21.00 + 160;; + 300 + 60 Control 110,000 Counts 110,000 1 #- 1 1021.22.11 Test compound - Benchlophen Test organism - .1220. mm Phenol coefficient dilution - 131300 Time of Exposure Dilution of Phenol , minutes 10 mm; 1.5 giants; ‘ 1-60 4- .. 1-70 + 7+ Dilution of Benchlophen ‘ l-1100 - - 14200 i - .- 1-1300 + - 1-1400 + + 1-1500 + + 1-1600 + + 1-1'700 + 4. w Test compound - Benchlophen Test organism -- @5me 31299.29. Use dilution - Dilution 1-100 1-100 1-150 1-150 1-200 1-200 Control Counts >18150 (18200 Time of Exposure 1 minute 5 minutes 10 m Plete Plate Plate ‘ Plate Broth Cgunt Broth Count 4 Broth Cogt Cogt " 0 - 0 - 0 0 + 0 - 0 - ‘ 0 0 + 0 - 0 - 0 0 + 11.20 + 0 + 0 0 + 1000 + 10 + 0 0 450,000 308 ,000 W Test compound -’Benchlophen Test crannie- - W m Phenol coefficient dilution - 1:200 True of’Exposure Dilution of Phenol ‘ 2 ninutgg 10 minutes 1980 - ‘- I 1-90 + - 1-100 +» + Dilution of Benchlophen lélOO - ._ 19150 -’ - 1-200 + .. 19250 + + 1-300 + 4- 1-400 + + TABLE 52 Test compound - Dowicide C Test creams- - .22; m Use dilution - 1:100 Tile of'Emposure 1.minute tes 10 s [Plate I Plate Plate Plate Dilution. _B32§B‘§oun§ Broth count _ngtB*gggg§4_jygyg;lgggflL1 O 3 O 0 O 0 I 0 1-100 1» 50 q. I 1—100 + 40 + 1—200 + 90 + 1.200 + 80 1. 1-300 + 120 + ssoooIo + I 00000 19300 +' 130 + Control 585,000 Counts 610,000 Test compound - Dosicide C Test organism - tag. m Phenol coefficient dilution - 1:5000 Time of Exposure Dilution of Phenol minutes _ 10 minutes B: We; _ 1.5o I - - - 1-60 - - - 1-70 + 1- .. Dilution of Dowicide C 1.2000 -. _ - .- 1-3000 - .. “— .. 14,000 - .. .. 1-5000 1» .. .. 1-6000 1- + .. 1-7000 w + + 1» Test compound - Dowicide C Test organism - Eberthella $2212.13. #6 Use Dilution - 1-5000 lanufecturer' s recommended dilution «- Time of Exposure 1 minute 5 minutg 10 minutes Plate Plate TEE;— Dilution r th Count [Brita Count M Count 1-4000 + 20 - O - 0 14.000 + 10 - 0 .. 3— 1-5000 + 31:0- 4» 0w -- o 1-5000 + 180 + 10 - 0 1-6000 + 11.70 + 30 + 10 14000 1. 2390 + 10 + 10 Cl 15,000 19 ,000 02 30,000 I 20, 000 Test compound - Dosicide C 202141.59. Test organism - Eberthella m #6 Dilution of Phenol 1-80 1-90 1-100 Dilution of Dowicide C 1-3000 1-4000 1—5000 1—6000 1-7000 1-8000 1-9000 140,000 Time of Exposure 4 minutes 10 minutes 15 minutes _ + - - + + + + .. .. + + + + + + + + 2 + + + + + + + Test compound - Tetrosen Test organism - Eberthella M Use dilution - >1:10,000 <1:20,000 1 minute 5 minutes 10 minutes Plate Plate Plate Plate Dilution Broth Count Broth Count r0 __C_9_1_:_n_t__ r0 Count , 9 140,000 - 0 - 0 - 0 - 0 1-10,000 - 0 - 0 - 0 - 0 1-20,ooo + 830 + 39 + 0 - 0 1-20,000 + 1500 + 30 + 0 - 0 140,000 + 2190 + 360 + 30 + 50 l-A0,000 + 31.00 + 60 + 10 + 0 Control 120,000 Counts 1 240,000 All negative tubes were inoculated with 1.. pm. All tubes Time of Exposure showed growth at the end of [.8 hours incubation. Test commund - Tetrosan Test organism - Eberthella txphosg Phenol coefficient dilution - 1360,000 Time of Exposure Dilution of Phenol minutes 10 minutes 1 1-90 - 9 1-100 L + r + I Dilution of Tetrosan 140,000 .. .. 1-50,000 + - 1-55,000 + - 1—6o,000 l + .. 1—65,000 + + 1-70,000 + ,_ “fl + “ 1-so,ooo I _ + + f W Test compound -'B. T. C. Lot # 900 Test organism - tag. auregg Time of Exposure .11.. 1.53:: ganja—1339:5122 1.2.03“ 93% “a“ r: r—--°“. r-‘fi-w “3 +4“?- 1-1000 - 0 - 0 - I 0 - 0 1-4000 1- Inn. -fi 3500 - 20 - 90 14.000 + I In. < - 5'70 - o - 0 1.6000 + * Inn. + 11.00 + 370 .. o 1—6000 + Inn. 4» 1520 + 30 - 0 1-8000 + Inn. 4. 1620 T. 20 — 0 1-8000 + In. + Inn'. + I 290 - 0 Control 790,000 Counts 820,000 W TQLE 56 Test compound - B. T. C. Test organism - Staph. aureus Phenol coefficient dilution - >lz4000 (136000 Time of Exposure Dilution of Phenol . 5 minutes _ 10 minutes 15 minutes 1-60 - , - 1-70 + I .. 1-80 + . I v 4. Dilution of B . T . C . 1-1000 - - 1-2000 - - 1—4000 - - 1—6000 + ~0- 1-8000 + + 1-14 ,000 1- + TABLE 5'7 Test compound - B. T. C. lot #900 Test organism - Eberthella Mose Use dilution - 71:15,000 <1:20,000 Time of Exposure I 1 “% “1%.; rm? p122. rmtifit Dilution h_B_1_‘p_t_h_J Count I Broth MAM Cgunt mm 1-10,000 1- 0 - I 0 - 0 - . 0 1-10,000 + 0 - 0 - 0 - 0 ' 1.15.000 f 0 - 0 - 0 - 0 1-15 ,000 + 10 - 0 - 0 - 0 1-20,000 + 580 + 0 + 0 - 0 1-20,000 + I 420 + I 0 + 0 - 0 Control 30,000L I Counts . 19,000 I — r Test compound - B. T. C. Test organism -BBerthella tnggsa Phenol coefficient dilution - >1zl8,000 (1320,000 Dilution of Phenol 1-80 1-90 1-100 Dilution of B . T . C . 142,000 144,000 146,000 147,000 148,000 1.20.000 1.25.000 Time of Exposure I 2 minuteL 10 minutes W I + + .t. - + + Test compound - Emulsept Test organism - Sta Dilution 1-20 1-20 1-40 1-40 1—60 1-60 Control Counts TABLE 0 aureus Time of Exposure 1 minute minutes 10 minutes 0 minutes Plate Plate Plate I Plate Bro Count ,BrothICount , Broth Count Broth Count + 6000 + 2140 + 3300 + 6100 + 2160 + 1190 - 3500 + 5600 + 800 + 690 + 550 - 3000 + 810 + 1810 - 360 + 3700 + 2920 + 380 + 340 + 30 + 3220 + 490 - 420 + I 160 210,000 I 265,000 Test compound - Emulsept T§§§§ 60 Test organism - tang. aureus Phenol coefficient dilution -1:40 Dilution of Phenol 5 minutes Time of Exposure 1-50 1-60 1—70 Dilution of Emulsept 1-10 1-20 1-40 1-80 1-100 1-120 I ++++ Test compound - Emulsept Test organism - Eberthella M Dilution 1-20 1-20 14.0 1-40 1-60 1-60 Control Counts Time of Exposure 1 minute minutes “was 111 Plate Plate 1 Plate Plate F'B'Li'tp‘ Congo t Br: Cognt grim Coogt {grgfhgqflgfini + 0 -- 0 + 0 - 0 + O - 0 + O - 0 + O - O - ‘0‘ - 0 1. 0 - 0 -— 0 - 0 w 4. 0 - 0 - 0 - 0 L 76,000 Test compound - mulsept Test organism - Eberthella tmsa Phenol coefficient dilution - 1:300 Dilution of Phenol 1—80 1—90 l—lOO Dilution of Ehmlsept 1-100 1-200 1-300 1400 Time of Exposure . 2' minutes 10 minutes 15 minutes .. J .. .. + l - - + ~I-w + + - .. + + + V . ,0 .1? "v I 1; '- s 1 J 0i“ ‘. . ’fx . 4,, m .5. ft . E; )4; , 5. og‘;1$ :35 ~ 1 O J p o n I ' 1 I. 5.; fl. 5.. & v ‘3. t 5, . '1' e,__ i'i s M "a 1‘- .}P '1 is»... 5‘1. D‘ e t “ . . .. fives c... .91.”... Ppatd 13..-. l .a 1.. at. -?3-.F. .._‘ Iv...“ .. . 1... . I . duh , ,.../.PV... '1‘ . ( ‘ l .0 V \l . .5 I V . ...(. . n i. . r . ... . ‘wm . .IN . ”a. .2 . . :32... 6 . - u v e .. .. .. - L sl.’ . .r .. .w\l.. . . I J A .rv {vii .. . r .J I . A . *.Ua. O .l at. .‘I 1. . .r .. . m \ H L .. . . v. .v . (WWI): o :1. 53151.. . I. l- 3. ,7 want J .s 'ruf: . . 3.1,...“ 3.1.2.”..- o 0.. ..1- I. «I. ..n..uda. ‘1. .- We m.-. . r. J. 1A.... .0... r...‘.... - .. . .c . .. .9.— . . .11!” {.DN 0 JV: 4: .u. 1. ‘9 7 . y . 1. . .‘C‘. u 1": 8. “If. .fl 6 t :64 V. nknuthr...,$r..0.~ .. . .h .. ../ fl" \ ~. .. . .. .. r I o.\,.‘ .‘f L". 3.! .. v ...I.. .l I. . .h. 7. ..h9vx..¥\b/...1i .... a... .. v . . 1.... .. I a}. . ti; .1... rr ., I“... u x . . .3 . 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