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Transfer of infection by handshakes Mathews, Helen M. 1926

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TRANSFER OF INFECTION BY HANDSHAKES.,, BY HELEN M. MATHEWS, B . A . UB.C. LIBRARY CAT, met L£s6?- mi fa- ny? S V- <~s i? 7 • win f i \ TBAESFE& OF IKFECIIOK M HAKPSHAKSS l x Helen I . MMlMm, S^A, A T h e s i s s u b m i t t e d f o r the Degree of MASIE& OF ASIS i n the Depar tmen t of B a c t e r i o l o g y . THE UNIVERSITY OF BEITISH COLUMBIA A p r i l 1926 . TEAESFEE OF IHFECTIOE BY HANDSHAKES In a s e r i e s of experiments we have shown that organisms both non-pathogen io and pathogenic are t rans ferred from one hand to another in hand shaking , -a f a o t of great importance in the study of the spread of i n f e c t i o n . Technique The f o l l o w i n g technique was used in each experiment . At f i r s t (Experiments 1-8) c u l t u r e s of B. prodigioaup Of v a r y i n g age (4-10 days? were used, u s u a l l y the pure eul ture c o n t r o l of the prev ious experiment; but l a t e r , (Experiments 9 - 1 7 ) , the c u l t u r e s of B. p r o d i g i o s a n were, on the average , two days o l d , grown at room temperature on n u t r i e n t agar . An emulsion of the c u l t u r e was made in . 7 5 $ s a l i n e s o l u t i o n . l a experiments when pure c u l t u r e s of B. t u b e r c u l o s i s . 3 . tUghthflf Uft *nd 9. typfeftittt »er® used , emuls ions were a l s o made in .75$ s a l i n e . Because of i t s dry growth the B. tuberculosa i s c u l t u r e was thinned out to a smooth suspens ion by rubbing the c u l t u r e in a few drops of s a l i n e s o l u t i o n wi th a s t e r i l e loop a g a i n s t the s i d e of the t e s t tube. The sputum weed was obta ined from the 2 p a t i e n t the day b e f o r e the e x p e r i m e n t was p e r f o r m e d . J u s t be fo r e u s i n g , the sputum was w e l l shaken . T e s t - t u b e s , each c o n t a i n i n g a s t e r i l e swab were l a i d mouth out on t he edge of the bench . In t e s t - t u b e s were the emule ion of the o r g a n i s m to be u s e d , a l a r g e s t e r i l e swab f o r sw&bblne- the emuls ion on t h e hp.nd, and s e v e r a l t ubes of s t e r i l e s a l i n e f o r m o i s t e n i n g the swabs . A l l the p l u g s were then t aken from the swab-t u b e s and the hands made ready f o r the e x p e r i m e n t by t h o r o u g h wash ing w i t h warm w a t e r and s o a p . When the " g i v e - h a n d " was d r y , or n e a r l y s o , the s u s p e n s i o n was swabbed on the g i v e - h a n d w i t h a l a r g e swab and l e f t m o i s t o r a l l o w e d to d r y a c c o r d i n g to the form of the a x p e r i m e a t . The terms u s e d d e s c r i b i n g the c o n d i t i o n of the hand* have been, f i r s t , " d r y " ; s e c o n d , "dry to eye and m o i s t to f e e l " ; and t h i r d , " m o l 6 t " . By " d r y " we mean hands t h a t l o o k and f e e l d r y and do n o t f e e l the o o o l n e s s of e v a p o r a t i o n when swung t h rough the a i r ; by " d r y to eye and m o i s t to f e e l " the hands look dry bu t f e e l damp when r u b b i n g the thumb and f i n g e r s t o g e t h e r and f e e l coo l when moving in the a i r ; and by " m o i s t " , hands t h a t b o t h look and f e e l m o i s t . - 3 When B. n r o d i g i o s u s was used , the l e f t hand was the " g i v e " hand and the r i g h t the " t a k e " hand, i . e . the r i g h t and l e f t hands shook each o t h e r ; but l a t e r , when pa thogen ic organisms were used , the hand shake was from one person to ano the r , u s ing the l e f t hand of each so t h a t the r i g h t hand of each was f r ee to handle appara tus when necessa ry . Swabs were taken of both the "g ive" and the " t a k e " hands, j u s t a f t e r washing, by mois ten ing a s t e r i l e swab in s t e r i l e s a l i n e and then rubbing over the su r face of the hands t h i s was done to make sure t h a t the b a c t e r i a to be smeared on were no t a d v e n t i t i o u s l y p r e s e n t . Then, a f t e r smearing the "g ive" hand wi th the emulsion or sputum, swabs were taken from the middle of the palm up the mid f i n g e r (M^) before and a f t e r the hand-shake to make su re tha t the b a c t e r i a put on were p r e s e n t . Swabs of the " t ake" hand were made a f t e r the handshake. These - c o n s i s t e d of swabs of the base of the palm (P ), the upper ha l f of the palm a t the base of the f i n g e r s (Pgim each of the f i v e f i n g e r s ( 1 - 5 ) , and each of the f ive f i n g e r t ip s (T 1-5) . The swabs were c u l t u r e d on aga r s l a n t s , except those of B,. d i p h t h e r l a e , which were smeared on serum. Means of i d e n t i f i c a t i o n When us ing B.. prodie:ioSUB no s t a i n i n g was necessa ry - 4 -as the o o l o r of the B_. prp dl-g^psus c o l o n i e s i d e n t i f i e d i t . However when u s i n g £ , t u b e r c u l o s i s , B, d i p h t h e r i a s ftnd £ • t y p h o s u s , s t a i n i n g of smears from the c o l o n i e s was n e c e s s a r y , J£, t u b e r c u l o s i s was s t a i n e d by Z i e h l -E e e l s o n ' s me thod , B-, d i p h t h e r i a s by A l b e r t ' s , and A* . typhosus by Gram' s , Hanging d r o p s and a g g l u t i n a t i o n t e s t s were a l s o u s e d in i d e n t i f y i n g B. t y p h o s u s , (1 ) The d i s i n f e c t a n t u»ed was made up o f : -640 c o , commerc ia l a l c o h o l (95$ EtOH) 300 oo , d i s t i l l e d w a t e r 60 oo , commerola l H y d r o c h l o r i c (35$ HOI) 0 , 8 gms. HgClg The a g a r u s e d c o n t a l n e d : -1$ p e p t o n e - Bac to 0 . 5 $ KaCl - C . P . , 3 $ Beef E x t r a c t (Lemco } 1,5$ A g a r - B a o t o L o e f f l e r ' s serum was u s e d f o r d i p h t h e r i a . S a l i n e - .75?* NaCl ( C . P . } The experiments were so performed as to duplicate the chief varieties of natural conditions of the hands when shaking,- i.e. surface (a) dry, (b) moist; and the process (a) firm, (b) dragged, fiecord of Experiments In the preliminary trial and the first experiment with B,. prodieiosusf the hands were disinfected, after washing but before Inoculation, to kill the organisms found naturally on the hands. The cultures made from - 5 -the swabs were incuba ted a t 37^- C, In n e i t h e r i n s t a n c e was there growth of B. prQdlplosu.g or of sk in c o c c i , presumably because of the d i s i n f e c t a n t . Therefore in ths second exper iment we did not use the d i s i n f e c t a n t before the handshake a l though the c u l t u r e s s t i l l were Incubated a t 37# C. B_. prodlKlosus p o s i t i v e was found on c u l t u r e s of the take-hand made from the base and upper p a r t of the palm and the f ive f i n g e r s . For the handshake in t h i s experiment both give and take-hands were moist to f e e l though they looked dry . In the t h i rd experiment both hands were dry , the handshake was f i rm, and the c u l t u r e s were l e f t a t room t empera tu re . Cul tu res made from the top of the palm, the f o u r t h f i n g e r and the t ip of the second were p o s i t i v e for ,§,, p r o d i e i o s u e . Experiment 4 was s i m i l a r to Experiment 3 In tha t the hands were dry and the c u l t u r e s were grown a t room temperature but the shake was dragged. There was growth of JL, p rod ig iosus on c u l t u r e s from base and top of the palm and from the f i r s t f i n g e r . There was no evidence of t r a n s f e r in Experiment 5 with give-hand mo i s t , the take-hand dry , and the handshake dragged. At th is p o i n t s l i d e s and c u l t u r e s were made of the v a r i o u s co lon i e s found on the agar s l a n t s both from the -6-swabs of the hands before shaking and after. These included -1, Cocci - white pigment. Gram • ; 1-1-J- microns in diameter (probably Staph Albus) 2. Cocci - lemon yellow Gram • ; lj- microns in diam-eter (Staph Citreus ) 3. Cocci - orange, Gram •*• ; 1^ - microns in diameter (Staph Aureus] 4, Cocci - cream white, dark centre, Gram +; 1-1-i-r microns In diameter. 5, Cocci - pale yellow, Gram + ; 1-1-jj; microns in diame ter. 6. B a c i l l i - wh i t e , Gram + ; spores 1x3 micron, (probably B s u b t i l i s ) Therefore because of the absence of B. p rod ig io sus on the hands before i n o c u l a t i o n i t was thought unnecessa ry to make swabs of f i n g e r s and the t i p s of the f i n g e r s s e p a r a t e l y of the give and take-hands before shaking; and t h e r e a f t e r swabs were taken of each f i n g e r as a whole. Experiment 6 was a double exper iment . The f i r s t p a r t was a f i rm shake (A) between dry hands . Swabs were taken of the take-hand; then the give-hand was moifltened by ho ld ing i t in f lowing steam two or three f e e t from the end of the hose . A f i rm shake (B) between the moistened give-hand and the dry take-hand was made and swabs made of the take-hand. However there was no growth of B. p rod ig iosus except on the c o n t r o l c u l t u r e s made from swabs of the give-hand (M.M2M3), The seventh experiment was a d u p l i c a t e of Ho.6 and the re was t r a n s f e r B. p r o d i g i o s u s on the base and upper p a r t of the palm - 7 and the f i r s t f i n g e r in the A p a r t and the upper p a r t of the palm in shake B. Shake A of exper iment 8 was f i rm and both hands dry but i n Shake B the g ive -hand was l e f t dry and the t a k e -hand mois tened . Cu l tu re s showed t r a n s f e r from Shake A to the base of the palm and f i n g e r 2 on the t h i r d day and the top of the palm in a week, and from Shake B on the top of the palm. On experiment 9 Shake A was f i rm and both hands dry and Shake B f i rm and both mois tened . Transfer was made in Shake A to both base and top of the palm, the f i r e f i n g e r s and the f i v e f i n g e r t i p s , i . e . to every p a r t of the hand; in Shake B to both base and top of palm the f ive f i n g e r s and the t i p s of the second, f o u r t h and f i f t h f i n g e r s . The c u l t u r e used f o r experiment 9 was only two days old while those in the exper iments before had been four days to a week o l d . As the t r a n s f e r was so n e a r l y complete in experiment 9 we came to the conc lu -s ion tha t young c u l t u r e s were b e s t ; t he re fo re most of the p rev ious experiments were repea ted wi th the young c u l t u r e s - on the average two days o ld . A c u l t u r e grown for t h r ee days a t room temperature was used fo r experiment 10. The hand-shake was f irm with the give-hand mois t and the take dry . The c u l t u r e s made from the swabs were incubated a t 37^- C and no & n r o d i e i o s u s grew except in the o o n t r o l grown a t room tempera ture , which showed tha t - 8 -a l though B. p r o d i g i o s u s was t r a n s f e r r e d i t would not grow a t 37-g- C. There were a few cocci c o l o n i e s on some of the smears . In experiment 11 a twenty-four hour c u l r u t e was u sed . The shake was f i rm with a mois t g i v e -hand and a dry take-hand , but the hands had been d i s i n -f e c t e d before shaking . The c u l t u r e s on agar were grown a t room tempera ture . On the t h i r d day there was no growth except on the con t ro l on agar made d i r e c t l y from the emuls ion. Af te r a week the c u l t u r e s made from the swabs taken of the g ive-hand before and a f t e r shaking showed the p resence of B. p r o d i g i o s u s . Experiment 12 was almost i d e n t i c a l wi th experiment 3 , the only d i f f e r ence being a th ree day c u l t u r e i n s t e a d of ten days . The growth of JU prodig ioaua on the agar s l a n t s showed t r a n s f e r to both the base and top of the palm, the second, t h i r d , f o u r t h and f i f t h f i n g e r s , and the t i p s of f i n g e r s 1 , 2 , 3 , and 5 . Experiment 13 was c a r r i e d out wi th the same c o n d i t i o n s a s Experiment 4 with the excep t ion of the c u l t u r e - three days old in s t ead of s i x . Transfer was made to the base and top of the palm, f i n g e r s 1,2, and 3 and the t ip of the f i r s t f i n g e r . A two day old c u l t u r e was used in Experiment 14 i n s t ead of a n ine day c u l t u r e as in Experiment 6 and 7, o therwise cond i t ions were the same. Trans fe r was made in Shake A to the base and top of palm and f i n g e r s 2 and 4 and in Shake B to base and 9 -top of palm and f i r s t f i n g e r . Experiment 15 was s i m i l a r to experiment 8 with the exoep t ion of the age of the c u l t u r e - two days i n s t e a d of seven days, During Shake A t r a n s f e r was made to the base and top of palm. Al l the f i n g e r s and the t i p s of the f i r s t and f i f t h and in Shake B to the base and top of the palm to f i n g e r s 2 - 5 and the t i p s of f i n g e r s S and 5, Experiment 16 was a d u p l i c a t e of experiment 9 in a l l r e s p e c t s and t r a n s f e r was made to a l l p a r t s of the hand but the f i n g e r t i p s in both hand-shakes A and B, Experiment 17 was dragged, with the g ive-hand moist to f e e l and dry to eye and the take d ry , A two day old c u l t u r e was used and t r a n s f e r w»s shown by growth of B_, p r o d l e i o a u s in c u l t u r e s from the base and top of the palm f i n g e r s , 1 ,2 ,4 and 5 and the t i p s of the f i r s t and second f i n g e r s . In the nex t exper iment , No. 19, a week o ld c u l t u r e to of B. t u b e r c u l o s i s was used f o r some time and good growth was obta ined in a week or t e n days . Because of the dry g r a n u l a r growth of tuberc le b a c i l l i a loopful of the germ-mass was macerated wi th two o r three drops of s t e r i l e s a l i n e by rubbing a g a i n s t the s ide of a s t e r i l e t e s t tube with a s t e r i l e l oop . When the emulsion was f i n e , f i v e or s i x oo.. of s t e r i l e s a l i n e was added. This was swabbed on a well-washed g ive-hand and al lowed to d ry . Handshake A - 1 0 -was f i rm with botu hands dry . The take-hand was then washed and scrubbed with soap and water and al lowed to d ry . Both hands were moistened by hold ing in the flowing steam two or th ree f e e t from the end of the hose . Af ter the swabs had been rubbad over th© sur face of th© agar slants, smears were made and s t a i n e d by the Ziehl -Neelson method. The smears showed B.. t ube rcu los Is from the give-hand (M, ), from the base and top of the palm and from the f i f t h f i nge r of the take-hand in Shake A and from the top of the palm in Shake B. At the end of a week B^  tubercu lo s is c o l o n i e s were showing up and a f t e r ten days growth they appeared as t y p i c a l dry creamy c o l o n i e s . S l i d e s were made, s t a i n e d by the Ziehl -Keelson ne thod and they proved the growth of J . t u b e r c u l o s i s as shown by the c o l o n i e s in the c u l t u r e . Transfe r was made dur ing Shake A to the top of the palm, the fou r th and f i f t h f i n g e r s and the t i p of the t h i r d . During Shake B the base and the top of the palm, the f i r e f i nge r s and the t i p s of the second, t h i r d and f o u r t h f i n g e r s of the take-hand rece ived B_. tuberculoa i s from the g ive-hand. Experiment 19 was c a r r i e d out in the same manner as No 8, u s ing t u b e r c u l o s i s sputum obta ined from a p a t i e n t the day before . The d i r e c t smears made from the swabs and s t a i n e d by Ziehl-Heel son ' s showed t r a n s f e r to the second, f ou r th and f i f t h f i n g e r s and the t i p of the t h i r d dur ing Shake A and the f o u r t h - 1 1 -and f i f t h d u r i n g Shake B. A c u l t u r e of J5. d l p h t h e r i a e was u s e d i n a s i m i l a r manner to the two p r e v i o u s e x p e r i -m e n t s . The swabs were c u l t u r e d on a g a r but owing to the d i f f i c u l t y of d i f f e r e n t i a t i n g between B.. d, i ph t h e r i a e ooooold forms on a g a r and the s k i n ooco l on the same s l i d e t h e c u l t u r e s were t r a n s f e r r e d to serum f o r t w e n t y -f o u r h o u r s a t 37^ C and then smears made. These showed t r a n s f e r p n l y in shake B to the f o u r t h f i n g e r and the t i p s of the f i r s t , t h i r d and f o u r t h f i n g e r s . Two days l a t e r smea r s were made from the a g a r g rowths and t r a n s f e r was shown in Shake A from the base and top of pa lm, f i n g e r s I and 2 and the t i p of the s econd f i n g e r from Shake B - the b a s e of the pa lm, the f i r s t and t h i r d f i n g e r s and t h e t i p of t h e s e c o n d , B_. d l p h t h e r i a e was a g a i n u s e d f o r e x p e r i m e n t 2 1 , Shake A was f i r m w i t h b o t h hands d r y . A f t e r wash ing the t a k e - h a n d w e l l Shake B was made w i t h the t a k e - h a n d m o i s t to f e e l b u t dry to eye and the g i v e damp from h a v i n g the hand c l o s e d between hand s h a k e s , i . e . w h i l e the t a k e - h a n d was b e i n g swsbbed, washed and d r i e d . This gave a t r a n s f e r as shown by smears made a f t e r t w e n t y -f o u r h o u r s a t 37^- C to the base and the top of t he palm and f i n g e r s 3 , 4 and 5 i n Shake A and the base of the palm and f i n g e r s 1 and 2 i n Shake B, Sputum c o n t a i n i n g J3. d l p h t h e r i a e and s t r e p t o c o c c i was used unde r s i m i l a r c o n d i t i o n s as t he p r e v i o u s e x p e r i m e n t w i t h the t r a n s f e r - 1 2 -du r ing Shake A to the top of the palm f i n g e r s 2 ,4 and 5 and the t i p s of f i n g e r s 1,4 and 5 and d u r i n g Shake B to the base and top of the palm and f i n g e r s 2 and 3 . After 2 2 a week smears again were made and D E types were found in the c u l t u r e s from the t h i r d f i n g e r and the t ip of the f i r s t and a l so from the top of the palm in Shake B. S t r e p t o c o c c i were found from the t i p of the f i f t h f inge r in Shake A and the base of the palm in Shake B. For experiment 23 a day old cul ture of B^ _ typhosus was smeared on a we l l washed hand. Shake A was f irm and both hands were dry and Shake B was f i rm with the g ive-hand moiet to f e e l ( hand c losed dur ing swabbing of take-hand a f t e r Shake A) and the take-hand mois t to f e e l and dry to eye . No t r a n s f e r was made dur ing Shake A but in Shake B there was t r a n s f e r to the base of the palm and to the t i p s of the f i r s t and second f i n g e r s , of a Gram-negative organism, d o u b t l e s s B.. t yphosus . Feces from a typboid p a t i e n t was used for experiment 24, Shake A was f i rm with both hands dry and Shake B was f i rm wi th both hands moist to f e e l as in experiment 23. Gram nega t ive organisms were found in the c o n t r o l , made by s t r e a k i n g an emulsion of feces in s a l i n e on aga r , in the c u l t u r e from the give-hand and in c u l t u r e s made from the f i f t h f i n g e r and the t i p of the f i f t h f i n g e r in Shake A and from the second and t h i r d f i n g e r s in Shake B. - I I -Bo ib c u l t u r e s «ar« made from agar as4 the e o a t r o l t i l the o a l y o a l t a r e tha t va t a g g l u t i n a t e d by immune serum, faa abeeace af t r a n s f s r of 1 . tjchcang) cannot be e x p l a i n e d aad • • t e g to the laek of time tbo experiment eon Id not be aup l lmated . Shaking hands la erne %t tho most common easterns of oar c i v i l i s a t i o n . Bus iness aon meeting f r i e n d s , aeqaa ia -taaeoo ant bus iness a s s o e i a t o o shako hands many t l aoo each dap, P r o f e s s i o n a l aon and WOMB shako hands o f t e n through tho day and tome timet vbaa s o o t i n g a group af poo pi o a l i i shako hands many times 1ft c l o s e s u c c e s s i o n . femee, t o o , ohako hando a l though perhaps aot ao o f t e n at men. Of couroo there are p e o p l e , e s p e c ! a l l / of tho l eOer lag e l e e o , who seldom ehaka herds whi le ot work, out do ee o e t of working hours . Although not la ft fa r a f t handshake, c h i l d r e n o laop hende many s e n t l s t i than a d u l t s aad t h e i r hando ara more l i k e l y to be d i r t y aad are aero of tea la t h e i r mouthe - a f a c t that may holy to e a p l a t a tea preva lence af "contagious" d leeaeo among e a l l d r e e . Es t imat ing as an average one handehake par pereen per d a / , tho popula t ion of amerioa ( l£9 m i l l l o a } 1 weald / l o l d a t l e a o t l t d m i l l i o n handshakeo yag d a / . If oae y t r t e a t of *• t e l e * 106 ,710 ,6*0 ( 1 « 2 0 ) , Mexieo - 1 4 , 1 6 4 , 7 t t l l t t l ) | Canada - 6 , 7 8 6 , 4 8 3 ( 1 8 2 1 ] , -14-1 the population were diphtheria carriers and one half of them had virulent organisms on the hands there would be a half-million transfers of diphtheria bacilli each day by handshaking alone. If 1 to 2% of the population were 2 ohronic typhoid carriers and 10$> were temporary carriers -"JL. ty oho sua Itself as well as other faeoal organisms can 3 be isolated from the fingers of carriers" - there would be several million transfers of jJ. typhosus each day. However because of the intermittency of excretion every carrier has not virulent organisms on his hands, or, if he has, they are not always transferred; perhaps because, the number of organisms is few or they are not on the part of the hand that touches during the handshake. If the organisms are transferred they may not cause disease; first, they may not enter the body, second, they may not reach the part of the body suited to their re-production and third, the person infected may be immune. Chart Ho. 2 shows that the greatest number of transfers is made Jul the palm of the hand. This is probably from the fingers as shown by a study of the mech-anism of the handshake. The fingers are contaminated more 1, Ohapin- Sources and Modes of Infection. 2, Chap in- Sources and Modes of Infection. 3, Journal, Koyal Army Medical Corps, July 1913, p. 657. -15-often by excreta than the rest of the hand and hence they transfer,by handshaking, any adhering organisms to the palm of the take hand; but, considering body infection, the palm of the hand is. less likely to transfer infection than the finger tips. Owing to these factors disease is not transmitted as much as these experiments would seem to indicate. Handshaking seems even more important in the trans-ference of disease than the use of a common towel - the former direct contact between unwashed hand, the latter indirect contact between washed hands. The hands are used for everything - eating, dressing, work of all kinds and for the removal of excreta from the body. If then we shake hands and receive some bacteria, unless we are careful we shall have transferred in a short time the organisms to our mouths. On the other hand if «e ourselves do not keep our hands clean we may transfer infection from ourselves to others. Our hands are continually in or on our mouths. The number of people in a crowd who have their fingers in the mouth has been calculated, to be about 10%, If is often just a nervous habit or ibIMmg the finger nails. Another habit is that of moistening the fingers before turning pages. It is a habit common to all classes and ages of people at any time of the day, whether the hands are clean or not. It might be - 1 6 a f t e r shaking hands wi th a d i p h t h e r i a c a r r i e r or a f t e r a c a r r i e r has moistened h i s f i n g e r s to turn the l eaves of the same book. We do not r e a l i z e how of ten we put our f i n g e r s in our mouths u n t i l we have some organisms or d i s c h a r g e s on our hands and then we have our f i n g e r s to our mouth only to r e a l i z e that in doing so we would i n f e c t o u r s e l v e s , 1, Organisms have been t r a n s f e r r e d from one hand to another dufing handshaking in a l l oases (30 hand-shakes) except in the fo l lowing i n s t a n c e s : -a. typhoid feces were used (Experiment 24) . b, the hands were d i s i n f e c t e d (Experiment 1 & 11) e . the J., p .rodlglosus c u l t u r e s were incubated a t 37-g-°C. (Exper iment ,1 , 2 & 10) . 2 . The t r a n s f e r of organisms was as good i f not b e t t e r when the g ive-hand was dry than when both hands were mo i s t . S. The g r e a t e s t t r a n s f e r was found on the palms of the hand. In fewer i n s t ances on the f i n g e r s and l e a s t of a l l on the f i n g e r t i p s . -17-P^ - base of palm P - top of palm 1-5 - fingers T - tips of fingers APPIVDII D t t a l l a d j a c a n i Mtffifi I&Ul i - C n l t a r e - 10 d a / s a l i a 1. t. S. • • 6. e. e. t. Hands was had with soap and watsr, than disinfectant, dried by waring owar gas Jat. Swabs taken fro> left hand - fire. Swabs of right hand - take. Swabbed glwa hand with broth on 1 tare of B, prodlglosas. Shook handa. Swabs taken from both fire Mid take hands. Hands washed and dlslnfeeted. Swabs of be th hands. Swabs rubbed over surface of slant agar and Incubated at ff^ C. RESULT -no growth. Experimaii t , ,^ - O o t o b e r £ 8 t h , 1 9 1 6 , 8 a l t a r e sowa Ootobar 2 1 s t , 1 9 2 6 , *a is ie S. 6 . 6 . e. B. f . Hands —S)aMJa wi th L i f e b u o y S e e p , d l s l a f e e t e d (Mid w a s h e d . Swabs taken o f c l e a n ( T | haade , f . , ? . , l - E i n o l , , '1^-5 i n e l . i e e d l e g r e s ) r f tge-V»1TS hand swabbed w i t h e a e l s l o a e f 1 . g r a i l -g l f l f y , | end d r i e d l a a i r a a t l i head d i d a a t feel d amp. Swab ef give hand ••Aire shaking -M^- aid pal a ap aid finger. Shake - fIra- dry. Swabs ef give hand as la • 4 I . Swabs Of take hand P , P , 1-8 f T . Haade washed, dislnfietal and wesfcel? ftfrttsl e r a r f l a a e . Swabs of both g l e e and take haade ? . , P - . 1 0 , Give hand swabbed w i t h e a r a l s i e a e f J^. o r o d l -£l£LULa* 11. Shake - aoist though not wet p flra i i . 12, Swab of give hand as in # 4, 13, Take hand swabbed ?lt P£, 1-5, T1-E, 14, Hands washed, disinfected and washed, 15, Swabs taken of both give and take hands Plf P2, 1-5, Ti.5. 16, Swab» streaked over surface of slant agar and cultures Incubated at 37^ C, RESULT - no growth, November 2nd added two drops of very dilute (HH4J2S to neutralize HgCl2~ no effect. .Experiment 2 - November 3rd, Culture sown, Ootober 28 th. Procedure - same as in Experiment 1 1, Control by awabblng suspension on slant agar, 2, Hands washed with Sunlight soap and well rinsed, 3, Shake - firm and damp. 4, Hands washed and disinfected, BSSOLT - £4 hgurs, at, 37# C, Control - good growth, slight pink color, 1, prodigiosus on Plt 2. 1. prodlelosus doubtful on ?lt 1,3,4,5, Skin cocci on cultures of clean (?) hands and odd ones mixed in with £, prodiclosua on slants* 48 hours - 2nd, 24 hours at room temperature. Control - turning red, B, orodlgiosus in Mif M2, Pi, P2, 1-5. Experiment 3 - November 13th, 1925, Cultures sown on November 3rd, Procedure. 1. Hands washed and sorubbed with Sunlight soap. 2. Shake firm and dry. 3. Hands washed with soap and water. iii. RESULT - 48 hours at rooa temperature, 1. Prod,lKlosuj in lllt ME, P2, 4, T2, Experiment 4 - November 19th, 1925 - Culture sown on November 13 th, Prooedure, 1. Hand washed and scrubbed £, Shake - dry - dragged 3, Hands washed with soap and water. RESULT - 48 hours at 21° 0 B, Prodlgiosus in Mx, M2, P1# P 2, 1, Experiment 5 - November 23rd, 19 26, Culture sown November 19th, Prooedure, 1, Hands washed and scrubbed 2, Give hand - dry to eye, moist to feel - Take dry 3, Shake dragged RESULT - 48 hours at 21° C B. prodlgiosus only in control, U± and M2 Experiment 6 - December 2nd, 19 25, Culture sown, November 23rd, Procedure. A, 1, Swabs taken of c lean hands . Pi P2 1-5, f i n g e r t i p s taken in with f i n g e r s , 2, Swab give hand before and a f t e r shaking Mj_, M« 3 , Shake - f i rm, dry 4 , Swabs of take hand P 1 , P 2 , l - 5 , T 1 - 5 B. 5 . Swab give hand as in # 2 M3 6, Shake firm, g i v e - moistened with cool flowing steam, take dry 7, Swab of take hand P i , P 2 , l - 5 , T l - 5 8, Swab of washed hand as in # 1, RESULT - 48 hours a t 21° C J . profllftlPSUS on Hj, , ME ? ? fta.Yfr B.prod lg iosus on A-M^, M2 B-M3 i v . Experiment 7 - December 7 th , 1925, Cu l tu re sown, December End, P rocedu re , Same as Experiment 6. RESULTS - 48 houra a t 21° 0 . Control r ed , 4 days _JL* prod,iKiQBttf. A- i^ .P P 1 Experiment 8« December 14th , 1925, Cul ture sown December 7 th , P rocedure . Like Experiment 6 exoept,-A Shake firm and dry B Shake firm, give- dry, take- moistened with cool % steam. RESULTS - 48 hours at 21° C 1. orodlgiosus in control Mx M3 ? 5. days 1. prodielosus in A-M^, P , 2 B-M3, 2 \ gajUt B. pro dialogue in A-Pg Experiment 9 - December 16th, 1925, Culture sown December 14th, Procedure. A Shake - f i rm, both hands dry B Shake - f i rm, both hands moistened with cool steam RESULT - S davs a t 21° C B. prodiglQSTifl in A - M l t P l f P j , 1-5, T1.5 B-M 3 ,P l t P 2 , 1-5, T2 T4 T5 • . Experiment 10 - December 19th, 1925, Culture sown, December 16th. Procedure. Shake - firm. Give - damp, Take - dry RESULT - 48 liours at 3?$ °C Control at 21° 0 and at 37£° 0. 1. orodigJosuB In control at 21° 0. Experiment 11 - January 8th, 1926. Culture sown January 7th. Prooedure. Hands washed, disinfected and washed Shake - firm. Give - moist, Take - dry. RESULTS - 3 days at 21° C JL. orodiglosua in control 1. nrodlelosus in Mlt M2 (feeble) Experiment 12 - January 12th, 19 26. Culture sown, January 9th, Hands wa Prooedure Hands washed, soap and w a t e r , RESULTS - 48 hours a t 21° C. £«. MJL&i&JLftiaJ in o o n t r o l M l t P l t & P 2 ? 8 days 1 . FrQflE#PStiff in P i , P 2 2-5 T i _ 3 T 5 Experiment l g - January 15th , 1926, Culture sown January 1 2 t h . Hands washed w i t h soap and water Shake dry - dragged BESULT8 - 24 hours a t 21° C. J> p r p d i p l o s u s in c o n t r o l - p ink ? flayt 1 , p r o d i e l o s u s in M2 P x P 2 1-3 T1 • 1 . Experiment 14 - January 2 1 s t , 1926 . Culture sown, January 19 th , Hands washed w i t h soap and water A Shake f i rm, dry B Shake f i rm, Give - m o i s t , t a k e - dry RESULTS - 4$ hours a t 21° C A Pia&islaJL&i m k-m1 , ?x , p 2 B-M3 , Pi , P 2 , 1 9 A ay «. B. p r ^ J K i p s u g in A-M^, Px , P 2 2 , 4 B - P L P 2 , 1 Experiment 15 - January 22nd, 1926. Culture sown, January 2 0 t h . Hand washed w i t h soap and water A Shake firm - both hands dry B Shake f i r m , - g i v e dry, take - mois tened with f l o w i n g s team RESULTS - 8 days l.PfQ&.ifiiQStta in A-M l f P x , P 2 , 1 -5 , Tx , Tg B-M3, Pi , P 2 , 2-J5,T3 , T5 1 week »• FfQdAfflftmff in B - 1 IiTfiffff\*ent 16 - February 5 t h , 1926 , Culture sown, February 3rd . A shake f irm - both hands dry B Shake f i r m - both moistened w i t h steam. RESULTS - 3 days a t 21° C J . pro d i g lo SUB i n A-M l t Px , P 2 , 1-5 B-M3, ?i , P 2 , 1-5 Experiment 17 - February 8th, 19 26, Culture sown, February 6 th. Shake - dragged, G i v e - dry to e y e , moist to f e e l , Take-dry. RESULTS - 3 days a t 2 1 • C. B. prpdlftioatts in II x , P x , P 2 , 1 , 2 , 4 , 5 , Tx , T2 v i i . Experiment 18 - February 10thf 1926, Tubercle Bac i l l i Culture sown, February 3rd, Procedure, 1, Fine emulsion made by rubbing culture and few drops of saline against side of test tube with inoculator then adding 5 or 6 cc. sterile saline, 2, A Shake - firm - dry 3, B Shake - firm. Give - moistened with cool steam, Take- dry. 4, Hands disinfected, washed and disinfected. 5, Swabs rubbed over surface of slant agar. 6, Smears made by rubbing swab on slide, RESULTS - 2 weeks 1, Smears, tubercle bacilli present in M,,P2»?i, 5, M3 2, Eye and slide, Tubercle bacilli in A-M^Pg, 4,5,T3 B ?l.p2»1-5» *2-4 Experiment 19 - February 1.2th, 19 26. Tuberculosis sputum from February 11th, A Shake - dry, firm B Shake - both hands moistened with steam, firm pro-longed. RESULTS -1, Smears - A 1 - 2, 4,5, T3 2, Culture and slides Experiment 20 - February 24th, 1925, B. diphtheriae AOD, Culture sown, February 23rd. 1. A Shake - firm - dry 2. Take hand scrubbed 3. B Shake - firm both hands moistened 4. Swabs cultured on agar. Transferred to serum on February 25th, RESULTS - 1, Smears - A-Mi , Px , 1 B-? 2. Serum A-Mi, Pi, P2, 1,2,T2 B-Pi, 1,3,4,5, TX_4 viii. Experiment 21 - Maroh 3rd, 1926. Diphtheriae ACD. Culture sown, March 2nd. 1, A Shake - f i r m , d r y 2 . Take hand s c r u b b e d 3 , B. Shake - f i r m - g i v e m o i s t , f i s t f o l d e d , t a k e m o i s t to f e e l , d r y to e y e . 4 . Swabs c u l t u r e d on serum, RESULTS - March 4 t h D i p h t h e r i a i n A-M l t P l t P 2 , 3 -5 B - P x , 1,2 E x p e r i m e n t 22 - Maroh 5 t h , 1926 . D i p h t h e r i a ACD sputum t aken March 4 t h , P r o c e d u r e 1 . A Shake - f i r m , d r y 2 . Hands s c r u b b e d 3 . B Shake - t a k e m o i s t , g i v e m o i s t , f i s t f o l d e d 4 . Swabs o u l t u r e d on serum. RESULTS -24 h o u r s D i p h t h e r i a i n A-Hj., T4 B 3 3 days A P2» 2 - 5 , T i , T4 B P l s 2 , 3 E x p e r i m e n t 23 - March 9 t h . . Typhosus - C u l t u r e sown, March 8 t h . A Shake - f i r m , d ry B Shake - f i r m b o t h m o i s t to f e e l RESULTS - Typhosus i n A - Mj_ B - P i , T l t T2 Experiment 24 - March 17th, Typhoid feces obtained March 16th. A Shake - firm, dry B Shake - firm, both hands moist to feel RESULTS - A -B -

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