\ ’1 -. Is’ . u A ([93 u 1..I YO; .u a .1 I. .l . I .I‘u If..‘. ..3IIIII, ultquo.cla.1\a‘tou4-.\ - $.31...‘ :14. ‘ _. ‘ ._ .r: {E altfil... I a k. .1 of» '4 r A y a ,‘ (0 .9? a A ,f l! . ‘ N \ a s I “x \ an n A a .. .\ . . ‘ A or! . ‘ .. a ; .1 ‘ , bk \ r . . L '1. '. I. a 5 u n. v 1.951} I. r . 3‘ at I. If X. n , PLACE IN RETURN BOX to remove this checkout from your record. To AVOID FINES return on or before date due. MAY BE RECALLED with earlier due date if requested. 6/07 p:/ClRC/DaleDue.indd-p.1 A STUDY OF THE BA TERIAL FLORA OF THE FECES AND IETESTIHAL CONTENTS OF THE FOWL. Thesis Submitted to the Faculty of Michigan State College of Agriculture and Applied Science in partial fulfill- ment of the requirements for the Degree of Master of Science. Mark Wirth Emmel a ;7 June 1930. guests I. II. III. IV. V. VI. VII. VIII. TASK-*3 CF COI‘FTEZTIS. Introduction ................ 3 Historical ----------------- 4 The bacterial flora of the feces of healthy mature foals -------------- 6 The bacterial flora of the feces of two weeks old healthy chicks -------- 10 The bacterial flora of the intestinal con- The tents of ten chicks affected with pullorum disease ------------ 12 bacterial flora of the intestinal con- tents of twenty adult birds affected with enteritis -------------- 15 Cellulose decomposition -------- - - 18 pH of the feces of normal adult birds - -'-— 21 Discussion ................. 22 Conclusions --------------- - 25 Bibliography - - - - - - - _ a ....... 27 q {LC' :53 U‘d 'u I. INTRODUCTION For many years it has been known that the intestin- al tract of man and animals is the habitat of millions of micro-organisms. Comparatively little work has been directed to the intestinal flora of the fowl especially in-so-far as a systematic study is concerned. many work- ers have reported various organisms as occurring in the feces of the fowl but none have attempted a study of the percentage occurrence of the organisms present. It appears that such a study would be of extreme importance as enteritis, apparently due to many factors, frequently occurs. The fact that this condition often becomes chronic would lead one to believe that the bacterial flora of the feces and intestinal contents of the fowl may be important in such cases and may throw a new light on certain factors which have here-to-fore been over- looked. II. Historical Kern (1) found twenty one species of bacteria in the intestines of twenty four birds investigated by him. A common characteristic of the bacteria isolated was their ability to liquefy gelatin. A relatively large number reproducedby spores. Only a few pro- duced color and these were principally micrococci. Bacillus coli was the predominating organism. Although Schottelius (2,3,4 ) did not identify organisms of the intestinal flora, he found that the presence of micro-organisms in the intestinal canal was necessary for the normal development of the chick. He concluded from his classical experiments, in which chicks were hatched, fed and grown under strictly sterile conditions, that the micro-flora of the digestive tract assists in the resolution of food con- sumed, that substances which stimulate peristalsis by then» irritating action on the intestinal wall are produced, and, that the normal flora destroys and prevents proliferation of pathogenic organisms which gain access to the intestinal canal. King (5) found the flora of the intestinal mucosa to be to a certain extent constant. This flora appears to depend upon the environment and varied within cer- tain limits according to external conditions. Bacillus coli was the predominating organism and it was almost always found in the ceca and colon, frequently found in the lower portion of the small intestine and rarely found in the duodenum. Gage (6) found that the intestinal flora of 48 healthy fowls varied to some extent with the conditions of environment and different stages of life. Bacillus 0011 was the predominating organism. Few obligate an- aerobes were present. Rahner (7) found Eggh..ggli, gram-positive cocci, molds, Bacillus megatherium and lactic acid bacteria in the intestines of hens. He thinks that Eggh.‘ggli is a constant inhabitant and tends to crowd out other organ- isms whose nature depends to a great extent upon the food consumed. Eggh._ggli increased in number as the cloaca was approached. Henes and Rochlin (8) found the flora of hens to contain but few species of bacteria and to be identical in all parts of the intestines. The uniformity of the micro-flora is explained by the presence of energetic acid builders. Thistheqthink. also explains the absence of putrefactive bacteria and their associated decomposition processes. III. TIE BACTERIAL FLORA OF THE FECES OF HEALTHY MATURE FOWLS. PROCEDURE. Samples of fresh feces were collected in sterile test tubes from the cloaca of apparently healthy birds. A portion about the size of a pea from each sam- ple was transfered to a tube containing 5 c.c. of sterile nutrient broth. The mixture was thoroughly emulsified. Dilutions of 1 to 100, 1 to 500, l to 1000, l to 10,000, and l to 100,000 were plated on nutrient agar medium. All samples were diluted and plated in duplicate. The plates were then incubated for 48 hours at 37°C. In most instances all of the colonies were picked from.the plates. In certain instances when desirable plates were‘ not obtained all of the colonies were picked from a representative portion of the plate. Transfers were made to slants of nutrient agar and incubated for 48 hours at 37'C. The slants were then replated and in- cubated for 48 hours at 37'C after which the organisms were transfered to nutrient agar slants. These slants were kept as stock cultures, transfers being made from them for work on identification. All of the colonies growing aerobically on the original plates were marked. These plates were then placed in anaerobic jars and incubated for 7 days at 57.0 in order to isolate any obligate anaerobes which might be present. The organisms, after being obtained in pure culture, were first stained by Burke's modification of the Gram stain (9) and streaked on nutrient agar slants in order to study their morphological and cultural characteristics. Bergey's manual (10) was followed in a large measure in identifying the organisms although frequently it was necessary to resort to other sources ( Weldin (11) and Ford (12) ) for a more detailed description of specific organisms. 'Acid and gas fermentation studies were first made on dextrose, lactose and sucrose peptone water media. These data together with those already obtained were sufficient in many instances to place the organisms in their proper genus. Growth on mannite, maltose, arab- inose, raffinose, dulcite, dextrin, salicin, rhamnose, xylose,and sorbitol peptone water media as well as tryptophane_broth, litmus milk and nitrate solution were very useful in identifying many of the organisms. However, it was necessary to resort to other media for the identification of certain organisms. The acetyl-methyl-carbinol test of Voges and Pros- kauer (13) and the methyl red reaction as described by Clark and Lube (14) were used in differentiating between the genera Escherichia and Aerobacter. There proved to be a very marked correlation between these tests. This corresponds to the findings of Levine (15), Clark (16) and others. RESULTS. The occurrenceof Escherichia coli in the feces of these hens varied from.5.4 to 78.3 per cent, the average occurrence of this organism.being 33.66 per cent. Escherichia communior varied in the feces of the same hens from 3 to 78.8 per cent, the average occurrence of this organism being 31.02 per cent. The occurrence of these two organisms collectively rarely fell below 40 per cent in individual birds. Escherichia neapolitana and Escherichia 533g; lactici occurred in the feces of two hens while Escherichia alcalescens occurred in the feces of but one bird. Staphylococcus citreus occurred in the feces of eight birds. While this organism.was not present in great abundance it occurred more frequently than any organisms with the exception of Eggh.'ggli and Eggh. communior, its occurrence varying in individual birds from 3.1 to 20.4 per cent. 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O a U . 9 i 0 c u g. _. fl "' - 3‘ r ' l . Q a a g ¢ Q, g e o I! 0' e l 9 S 5 I: a 1 , a 4 4! B. ClI'CluifiElno o a o ., o o v -‘ a . o ., : .~ 5 .. .3 ‘f - — - ”nuns-'- “« ‘_ nu -. _' » M v-mwm.“ —-.- a. ‘-\v-- ~o-q -—-... :- -_‘-‘.A~- 4-"... mw~vaw w m _" *rlrwwru -.-r'-- u. .W-c . u - ‘ “'3. era-v v. 4- *WO-a‘ —- V. HI. V ~ .-,.... v... "“0“ a o c- o o n '3 a a s .~ - A A .. Q ~’ ’_ . 1 3' (J '.- € ‘0 (' 1V 1‘ 3 q} , 4 -, \ ; _\ 7‘ 'o 3 ~. ‘ w“ J. 7 o a a s ‘ ;_ ‘ {-w (-1 . 5/ 1 1 Q n (I (I f t A , . O "‘ ‘- 1 a '0 a ‘ ' .Lj . 1216‘ ,C14 baler]- #511,]. 9 0 o t c T ‘ " " ' C 3' ' a 2 a a 4. 2 9 4 “.4...— . --»_- _. up— —. w..— _. -.-.-a.. -——-- w —1~ ' m ”m *w‘- .-. I- v v >- - -. ...' 4M,”_W~ I ‘- I‘ .“ amb" ~ “‘ ’ . a a n .l ’ I. O 0 O ‘I R '7 a g g a fi 3 a 2 v.- r: :1 r3 ‘3 “ " ° 3 7 ' q ‘ ‘ " Z ' ' ' A 1" . 1 . . . ~ . a Q g. o s O O a 6 —. r n A l a ‘LA , . » ‘ ' V ’ " 3 ‘ II 9 n .:-.C...-.I'O.CIO. --lCLLU.Ia.Cl€flS . a v ‘ L -‘ 9 . e *1 04 " " 3' ~ 0 7- Uo . 3 a 4' ‘ r .. _ m ..—-=-—..-r.~..-s. .‘--. H»... ——-—-— — . v — no “wt-"~- 4m”... .- -.---o~“r . ’vw-v—v-o- .- ,, 4 ... _. _ ‘ »-m‘ «awn-Wm“ g p a o o n a m ; c a o n a g a ., a "’ c a a .' s n o e A. g o a; g .. a , 4. .. .3 a t j 3 , + ‘ ' n o o’ e '1 a a I! 17"; (a o c o a o 9 2 , 'F n e» c 3 a ‘1’ AC .1 0 I511 C - 0 flag tip.) a u c- o v '3 n c e- o 0 O - 5-4 Q ‘\J s; ’3 r. v, '5 . f 1: - -, "v la- < »- m7... .-.‘- c m -... gnu—.4 -‘ —-— '— —. H .4.- "—0- "Jo‘s--v — __ 1 -“" ‘. -"~“I"'w"t- a. 1...‘ --.w ‘ c v. ~-—-v-I.- Mam-'- “1..“ p a n o a o o b 9 o T " ‘3 4‘ e 4- .p ~ .. c m c. a k r. 9 I: r: a a " " "‘ ’ 4 a ‘ I : B t \ . 1 a o o a a Q a a - a a I: 7 O n ‘ a a A. a _. _ . V . _ . . a . . 13188613 LI‘lCdS a a. . r c c o a . , do n .- - ». > a. a 3 _ a. M -so~-.~ Mt. A ._ - - ‘Aanvn‘ “ma—p v-. r...“ J. .. . ——-—V__._._._ o v o 1 z- .o 9 o o a ‘3’ “ 5 —» a, . a «an .9 9 A, g A _. . e . l a . u u . r ‘ . . , '5 o f‘ B cereus - « - - » . , . . . m : " o 9 ’ “ * 3 7 ° ~ - . fl « O a u o 9 a p o .. a ., ' . ’ '7' f O i‘ . ,. L . o u w — » .Q - a. wow we. ‘ x.‘ 1' v—v— -.—_ i . e .._.. _ a c n a o c a o f a . , g z " " ° “ a n a a a o I: o o n u o o ' a o , ° 0 “ '9 ‘-‘ I, 7 7 r . “I“. - ' . I A ’ D 1e 381 as - . . . . . . . . . ~ 9% a . . . 0 I o e a . ,9 J o 49 g Q .5 n n . 2 D a A U 5 n - J \ . N a ’ 3 a . ~ L I _ «n- . a fig. w — _ rv— owl. “I“. m o 9 Q or o o a a 0 Q .0 o g . . . a a o a O O p a a z a o o c . e 9 5 o u a C O . Pct ruber ° ° - ‘ ° ~ ' 4 a - e , - - . . . . g ,, g n c c r - 9 a O o o ' ° ‘ ‘ . o . 0 o a s .1 a a n 9 o o a a C o n- a O o g g Q . 0 o o o o .- 6 u» o c 9 n O o o g ’ 0 I ’ 3 ‘ C D O . fl t ' ‘i 1 ' '1 o ., ,. g . 9 n a . ‘ ¢ 0 - _ - . . 5 5 . . . . Lac 0. aCicopiiius . . . . . . . . a . . . . .H . . , o g g : p o a. g C O W: a A.» ’. . ,ln 0 o g .. . . ._ . ‘4 D 3 I O O O O O ‘ t. O . . . G C O C _ O f . C .. S s 0 O o o .0 o o .u o a 4- .. o a 3 ., 4 , - 6 G O ramo U. 9. . a . . 4 a ._. . o a I o o d k I C 0 . . o a q. . g . . . ’ ‘ D a o a . . a,“ --—.... 4‘ . C . O Q 7’ Q . . . O C C D o o o o a o 9 g 9 a Q n a . ¢ . o o a t M ' O 5 fla ' - ' G O 0 0 a D O II 1cr . Ub vus . . . . . . . . . ' . ° . ‘ “l l ' ' ° ° “ * o D o n c ._,' . e a u o o 9 ‘ . —— ——7 D o a a o a g Q . . . O o c a o n n 4 . . . z : . 0 e o o c a haer ” ' “ “ * - a B O C o ens . . O n a a g 0 . o g ‘ " 0 ¢ 9 o a a a '7 .. x O , Q g . g ’ O \ ,. g 0 , 0 l o 9 c U . c e 1; n o 9 4 . o O n A. O n C D t b b' ‘ ’ ’ ' ’ ' ' ‘ a - a z z 2 r 2 ~ ° . . . AC o O l 1 . . . . c . . . 9 M 0 9 4‘ Q ' 9 o m ’ ‘ a I s y o 9 : = : 3’ o o a P n O n a . o . I, ’ p o J 5 d J . A) D IO ‘ ,3 U ' 9 I 0 n n a " m B vul atu ' ' - ' . - . : : . z . ~ . - - . . - . . . s . . . l '9 u n a c O a 0 n o a o o a o ‘ m 9 . . 0 A. Q . : . . O O 0 Q Q q 0 g Q a Q . o c- . o o o o 0 L4 . 0 c e *2 . 0 ’ 0 o a o 0 ° ° 9 ' ’ ‘ a O l . o o a . p . . 0 o a a «a o a a O l 3 c . D s q a n I, 9 0 a "' 9 0 o ' O . . Q Q 3" O Q C “‘— 9 . . n l . . C Q Q . D . 9 . Q‘- ‘ O , . 9 o a; 5 1, r G v O 0 . 9 J 'J o .0 0 v. .. . . C I. Q . . V . ’ . " u . z o z 9 1' a a a g . . . O O C . . . " ‘3 5 O a Q . a l ' - OO ' ' * . '. 0 a o . . o o - - - - ‘ ' . 00 ~ 10 - oo ’ o ~ « . O V‘ C O D V 9 3 v '1 9 OI‘Sanism in each bird. Aerobacter aerogenes was found in the feces of but two birds and constituted, respectively, 5.1 and 6.1 per cent of the total intestinal flora. Two obligate anaerobes were found in the feces of the twenty birds, Bacteroides bifidus occurred twice while Clostr"ium sporogenes occurred but once. Achromobacter liquifacigns occurred in the feces of three birds. Salmonella icteroides and Actinoqyces micro- flavus occurred in the feces of two birds while the remain- -v .1. 0 in; organisms occurred UUb once in the feces of various An organism not identified occurred in the feces of three birds. It represented, respectively, 5.4, 38.3 and 36 per cent of the total flora. The behavior of this or- ganism'sas so nearly alike in each instance that it seems highly probable that all three cultures were identical. The organism constantly stained negative by the Gram stain but showed a very high per-cent of involution forms when grown on various media. Its motility and physiological activities on different media varied so greatly from time to time that identification was impossible. 10 IV. THE BACTERIAL FLORA OF THE FECES OF TWO WEEKS 01D HEALTHY CHICKS. PROCEDURE. The method of procedure was the same as that followed in determining the bacterial flora of the feces of healthy mature birds. RESULTS. The presence of :E_s_c_13._c_:_o_l_i and 13393;. 995g- munior in the flora of the feces of the baby chicks studied appeared to be rather variable. gaggg..gg;; was absent in the feces of two of the ten chicks while @533, communior was absent in the feces of one chick. Eggg,‘ggli.was most abundant in the feces of chick No. 4 where it constituted 74.1 per cent of all organisms present. In the feces of chick No. 1 80.7 per cent of the organisms proved to be Esch. communior. The average per cent of 32220.22ii found in the feces of the ten chicks was 26.3, while the average per cent of Eggh. communior was 55.01 . .§§320.§£i§£ lactici occurred in the feces of four chicks and constituted from 10.7 to 30.2 per cent of the total flora. Eggh. leporis occurred in the feces of one hick. Staphylococcus citreus and Staphylococcus albus occurred in the feces of two chicks while Staggylococcus aureus occurred in the feces of one chick. Organisms of the genus Bacillus occurred in the feces of many of the chicks but usually represented a small percentage of the total flora. 3, mycoides was found in the feces of six chicks, 3, mesentericus and o S. k m. r . 11a .0. O. 0. ~ _ C I. o. _ f— m ) a. no no oe w _ O (J 5 so on on o 6 l O - .... a. w l O ...-a on on ... M1 Wm w .o a. .54.... O t .9 Mu» .. .. a. .. .. i ._ ~ .. .. .. o. . m 9 1v“ 5 an on «o ..w “ fl . o ca .9 o. on o no .3 WI. .. .- .. .. _ 7 .. a. .. n on on no 9 "AW 4 so :4 a. o. _ _ 0 no on a. a; a 9 or at Cu 0 «no go... 0 o 0. on m .. ... a. .l... O . a. a. o. l .o p. o. 8 Op 4 .0 «a a. 70 n h on on... a. no on p 1 L _. .. : : . .. : : . -. : : : a ... a. o. o. . . o ... o o 4 no .— n .q .. .- .m». ”no.“ s E. o. ..~. A o o a. a. .. o w . o no a. .- .. .- H...“ k ... .. ca 9. r . comes an a. r . . so a. n. to V0 . . o. o. oo o. S c 7 ... ... so . a . no a. a. w a. go ... i o. a. on e 01 5 o. In. 6. G! v u n n WDO .0 0c .ofi 0. O. a. 0. w 9 1 I. O. O. o. .. h s. . H . . M 2" . 3:: . . 4 O n nu so .0 an 6 _ oafloo ouw .- . _ a r a o.» our. . 2 fl - an a. .9 o 7 2 O O. O o o at no. . u o l l o a... go on a w _ ma.» 00‘0. u“ u m _. ~ . Q on on o O o co .0 so 2 3 O . ::. . . .... . I. . , Z... 5 a ...: ... 0 e 6 l A. on no as o M a w .— mo 9... oc*oc .. m * PA.” on a. no a l . O 2 l #0 a. o. H [mm A10. . G 00 6° ’9 O. H “ r p ..¢ Av IO~OD c9 0 m w M. O ... O. O. .0 w Phi“ ‘ O. O. , l 9 09 _ . _ 0 00.90 . o e 3:... 4... c ..:: M . h. . _ ..::. a ... ......z... . o n . i 7.:_ . . , l a . o m ......— na go an an al.— m i «...... oama . MN n q . w on so I. a _ O O 7 a a. a. a. l 5 6 ‘J on 00 .9 l . ¢ N < 0 .0 .9 OK. « M m . O .0. '0... w 8 5 . O. .9. O. a . L .. : . . . .. . _ . _ _ .. .. 6 ::. T S O o. a. -. h . ‘ «Tao ...“. ~ h .- .90.!!!- 9 on m 1 41 a. o. .a _ . , ._ . « ...- : 9 n 5 . a no no 8 a. on 0. Au. a...“ nu“: ...... a + m _ . .. w o co as so“. 0 fin” 6...»... an. 0.. O .m no .u a” 4. o h a . oouoo new.» a m . a ... co ea c. 94 no co co off-bulk?! _ m.— a l a no a. a. _ M . ~ 0”: q.- Ou . O 0. no 04 ..Ob'htih Ow .. .- . o .. .. - . . i m _ .. -. i . . a .. .. .. .. . -4: A... a: a ..u a a . O ‘ a. I. O 7 r A.” . O O. .4 a . ...-n. D .0 . w 0 . . I O. .O at O. 7 0.” O 9 .84.. on“ a w . ~ . O on“.. O .o l 8 fix“ 9. O. .0 .0 O .9 on no a: ~ 0 M .on our: on. w m a U 4 . owoo no .9 .. -. 0.9 a. on a O f o_ : : .. .. . Wu: F l M ... i... s; w m . aw“ 3... :4: : .. : . : : : : l O 5 N 8 w t as a. no w a M . F .59 o. as an. ~ 1 . a .v w .9 09 O! O. O. .9 _ e a . .. .. .. .. h . . _ . L .72....” 4 9~ . m m .. .. ...: , _ : 4 0 .. .. : _ a m ._ .... ...:1 - . : : . 0 _ m a. no a 4 W . on no 0 fi «.23. a 04. w as oo o w a a no a. a O , 0 on 0 a , 3...? on n a _ -.. . a co 9. . o O. . e 5 O D 0. a. a. 8 “ M . QQ~OD a" w ” LW H M O. 00.30 Co“ « GO O. O. .00 O 1 r 2 a .. .. . . . . .... .7. . _ . . .. .. . .. WW. A” 0 no a. a... o n _ u » 7r” . a. an": « r M . .. a. o. co .0 o a a a h L . n o . .. C a. a. V: «.../N WWW _ 5 . 3“: ...... . “ flJo‘. RN, 1 . ... 3”: ..v. w m h .. o o. .. .. .o a m MW , no . _ . 9 on o . a m . c on. . . MW .0 no no to PM“ 00. fl . ... 69 C. 00‘. 5“ firm” A" m ~ m “’0‘. CD .0 0.“. t a. .0 O. C. O. 99 l .. .. .. . l 9. O4 ...: .. . .. 2 .... W . ... . . .. .. . v .. .. .. l 7 . .. .. 9... .. _ 5 .. r .. h . w a .. : . .. e . o 5 .. .. . r. 7. .2 .. o” .. :7. . . w .. . .. .. ... _ Wu on a. on 0 AU. 0“ Ru 5 o no go... o . / AM.” all... n _ . 0—.9 0.. as o a H g a w v .o a. n. oo o _ O +0 .. .. .. .. mu 1 A. Q” o A“ A. . .u: .. 2. .. F . a " fl . _ .... .74.. 3w a. " .... .. .. .. m n .o a. no on on on “Q 0 L w H .0. ‘0'... no". u . w r .- . o .o‘oo 49" o . .. an o. no a. u.- a on a... , _ . o o o . . . new on 99 on on mm H ~ M M “ ‘ ...": :T . _ v m m g g .o no no a. * M c. at . _ a e g. , On . MW _ .- ... on"... on“ .- M ~ ~ ~ .. . o...- .f. on so a m m on coma. 49”.. ~ MW a... as. o . a. x q o- n; a 9 H on . “DI..— 8 HQ ~ W “.0 onvou now a H r a ’ ... owns an no _ w r 0. onuoo on a l .l m... “ ... . r: ...... ...... . ... H . L . Z .. .. . _ H v . .. 2 .. S O C . a -. .oo co. m , F . to»... .l S e. 8— a H ..a 0.. , a no... ~ 0 .m n .. .. .. ._ . .. n . . i. i . .. .7 ... .. . . l m .. .. .. .. i m F0 .1 m .w u S m v 4 M a. W: . C. . .- m4 w m .. 4.. o. ...:- o. . 4 _ o. .. : .. . .. T. m .. ... .- ... a ..v. .. ...-a . T... N _ m m . m . 7.. . . l S a .. . __ . h c .- . WW MW 0 m u ....m e a H 5” Tc; 8” 4 C.” . _. C. u T 4 y W m .. .. ...... ..w 4 _ O C a .o d h SD 6 nut". 0vw .. 9.... S . mat: w 60 ~ 8. . m a.» 9. C9 to V O ,4 . . a i u t b. .-.: . l e . I. . . u. a n W . . fl : ... l _ an ... O o e O a S S .l U... M m n S. .M a O. ..4 6' _ _ “ ... .. .. 2 C an.- t h fl C ”U ..l S S“ n“ Hm e .l_ t. .. W S a m“ C _ . . .. .. S _ C p 10. VJ O ...u 4....” _ . S C .. T. «J. -. .l u .1 S S .u S n4 .... .0 a C .< m... n._ . .l. f O S e +0 8 T4 mug «MUN PW ha ...... r O a JD PI. ...): U C 0 NJ n u C e n O T _ S .1 _ 7.. ._ t r r 7. e . . . t v. r 3.. e. _ a _ r. a g . _ u c a 3 ...) . C . J. .1 a” P s eh d. .... 8 +2 u .0. C n S r; 7mg 8 .l . .... C hC l 4 C... l a .1 v r ”- 1U S S . a .1 fi 8 7.. L .. r m. f... h .. w . 3 m. c. w . l i n .m e m s . m , mm 3. a o. ...a . q... o l _ _ . f t e e t a i .1 . ...; (O. .3 o S T. .60.“ V m o .0 O O o .l M... ..Tu u l l S l _ C n -. s. ...-m e s. a . a .m 0 m o t w n 1 m m o w a -... 7.; u 0 O ..m r 0 d ....DL h 6— a a .1 l p a: 4 r: .. a... a r L S . 7* a e .l . i 7... .... t C 1 u o n 1 . u s _ 4/... o it! 8 m W “A 1.4. l a e t e u .L . _ OJ 7 C e C .l T C t ...:l A m I. a C r e or“ . cl 1 01 B” Dr 0 M T. +0 My." 0 C at l B S e . h . c S B . w t . ...“ DD .1..,J C t C 7..” A C J A 8 B 11 g, subtilis were found in the feces of two chicks while §_B_. lactis, E. Retasites, 3. flavus, g. vulgatus, E. terminalis and g, circulans each occurred in the feces of but one chick. Aerobacter aerogenes was found in the feces of two chicks to the extent of 4.5 and 20.4 per cent respectively. The remaining organisms occurred but once in the feces of the various chicks. V. THE BACTVRIAL FLORA OF THE INTESTIHAL COE- TENTS OF TEE CHICKS AFFECTED WITH PULLORUH DISEASE. PROCEDURE. A sample of the intestinal contents was placed in 5 c.c. of sterile physiological salt solution. In instances where no contents were readily available the mucosa was scraped with a sterile scalpel, the scrapings being placed in 5 c.c. of sterile physiolog- ical salt solution. In either case the samples were thoroughly emulsified and a procedure similar to that employed in determining the bacterial flora of the feces of healthy mature birds was followed. It is regretable that no record was kept as to the exact location from which the samples were taken as Kallmann (17) has shown that there may be considerable variation in the presence of Salmonella pullorum at different levels of the intestin- al tract. Sal. pullorum.was isolated from the various organs of the ten chicks with the exception of chicks No. 2 and No. 8. No attempt was made to isolate any ob- ligate anaerobes which might be present. RESULTS. The occurrence of Esch. coli and Esch. com- munior in the intestinal contents of these ten chicks did not correspond to their occurrence in the normal chicks studied. §§§§§° coli was absent in the intestinal contents of two chicks and varied in the remainder from 4.1 to 50 per cent, being present in an average of 10.9 per cent. An average of 2 per cent of the flora of the 96 90 . T 80 ' 70 - 60 30 percentage occurrence 80 10 Chicks 12345078910 Chart No. 1. Percentage occurrence of §g;.'§2%;gggg. represented in white, Esch. coli, represents n black, and Esch. communIor, efieded, in the feces of ten two weeE oId cHIcEe affected with pnllcrnn disease. 13 of the intestinal contents of ten chicks consisted of Esch. coli, this organism being absent in eight chicks and constituting 13.9 to 6.9 per cent of the flora of the intestinal contents in the remaining two chicks. -- There was also a marked reduction in the prevalence of spore F5) vanisms in tie intestinal contents 0 s y- . .- V spore bearing or these ten chicks as compared to the normal chicks studied. Snore bearers constituted an average of 3.9 per cent of .L. B. mycoides occurred in the _+‘“‘- the total organisms present. I. subtilis, E. intestinal contents of two chicks while ramosus and 2. fusifcrmis occurred but once in the intes- . 4. a : 1 .110 ten C-1-LC;CS. ti.al contents of There seemed to be very little chance in the prevalence ‘_) J. of Staphylococcus in the intestinal contents of the above chicks. Stash. citreus and Stash. aureus both occurred in the intestinal contents of two chicks while Staph. albus occurred in the intestinal contents of but one chick. An interestinr feature of the bacterial flora of the intestinal contents of these hicks was the presence of ‘ ~n- . 7a .C‘ .L"‘ . "‘ A L‘, a». OlumfllSdS oi the Cenera .bsitnella, :roteus Salmonella 1 (31.. J. . 8.310 L) o;“?v3 JOCOCCL‘E . Ch "‘1 *1 “' N La, “I 7 ‘- .e \ ‘1': "P ‘1 :Jal. ll 1 rtm1»¢as inn: QTCCJLllfl8AuJL3 orvxniism.i11 the . ’L IN . s n - J- . J- a e . e \ intestinal consents 01 all of the chicks with tae ex- am 1-; fl rim , _, . 4n .: - ' Coptic“ 0 one. ins occurrence of this organism ranged '0 a 1 ’7' W ‘. 1'3 7 .1. C‘ 4.1 . liom lo.l to 1%eu per cent 01 the total organisms present in the flora. "l'. ‘l’ r.‘ ., ’ [Lin-LI‘ so sojfio so so so so so so so so so‘ss so os so so so so sows. so so so so so so so so so so so so so so so.so so so so so so so so so so so so so so so so 9 2 . O o e 4 5 O l w mm _ s 1 1w L m _ so so so os so so so solo. so so so so so so so solos so so so": so so so so so so so so so so ss_oo so so so so so os so“ so 2.: so so so so so so wolv- so so 0 s 5 4 .9. o 3 s O 8 7 was 5 O .. .. _. .. .. .. .. .. .. .. _ _ 1 ... so so so so so so so so so so so so so so so so so so so so so sowss so so. so so so so so, so so_ so so so so so so so so so so 5 5 O F 8 O O 0 KW n/W l 9 O . O O O s . O 2 5 l m 3 N 7 2 l I so so T. so so so so so so so so so 2. so so so so so so so so so so so so so so so. so so so so so so so so os so so so so so so so so o“— oo so so so oo oo 1 6 J w, w 7 o,» a J J O i A. 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I. .0 8 Lb _. as n H t. t n H s e O .l S T .1 h... a. e 8 ..Tu a S n .l e P r C C .1 S a. C d S a m 0 a .1 r e ..b a .l G .l S .l m S S. l ..O a n S a: .1: f u n r .1 nm U .l U n 8 S. e .l u l S e si e e n U r t e i a l U S m e O i u i so . . S S l a .l m 0 C I ..O C S a U .0 O r r P V Ci 1 O .l e a a g l m «I. a u .l u u v l r 0 .Tu a O .l v.» om .Tu 10. ... V. r 0 O l l mm C 0 8 G0 a e f .l e b l 0 DJ .1“ .l .1 1 O c c u a O O i s a .l c n o ..e_ t O l s p e e m n; ..D u o S e e m e n . ..D C e u I O P h 0 O a «H... e h 0 u h 0 DA D. O p u I m e ,_, h h . e P h r r a t D, o f B h e e r e m s m _ ._ t i. C C l r a C h 0 a r a C r r h r I r O v s s a t t s c . . r t. e . t 8 tv t c t . . . M r E E S S S E A B T P S A. B S E S S A S mm B B E P v 15 VI. THE BACTERIAL FLORA OF THE INTESTINAL CON- TENTS OF TWENTY ADULT BIRDS AFFECTED WITH ENTERITIS. PROCEDURE. The method of procedure was similar to that followed in determining the bacterial flora of the intestinal contents of the ten chicks affected with pullor- um disease. It was necessary to secure more samples from scrapings from the mucosa as diarrhea was a com» mod symptom in these birds and the intestinal tract was in most instances devoid of abundant contents. No record was kept as to the exact location from which the samples were taken and no attempt was made to isolate obligate anaerobic organisms which might be present. RESULTS. There was a marked variance in the prev- alence of Each. coli and Esch. communior in the intestin- al contents of these twenty hens as compared to the flora of the feces of apparently healthy birds. Eggh. 23;; was absent in the intestinal contents of eight birds while Esch. communior was absent in the intestinal contents of six birds. Eggh..ggli represented an average of 12.2 per cent of the flora of the intestinal contents of the twen- ty birds. Esch. communior comprised an average of 10 per cent of the total flora in the same birds. E, cereus, E. mycgides,‘§. subtilis,‘§. megatherium, 2, cohaerene, Q. fusiformis and.§. ramosus occurred in the intestinal contents of the various birds and repre- ', - .h .9 * 0 -,.,~ 4. 1‘ L1 J 4-“ - 1 .. sentgd an A ere e oi o.J gcr cent or tue bObzl ,lolo. /’ .l I! '1‘ n ‘5 ma (“J-n \ 1a -n H n (— ~ 4‘ Stare. cu_eus ,nd no-lh. alous re3_ecenccc on ‘eisgc M occurrence in the intestinal contents of The presence of Stren. lactis, Strep. pitrovorus, q ren. Eyorenes and Stre . bovis constituteu an average t of 4.6 per cent of tne organisms present in the intestinal contents, occurring in 5, l, 2, and 1 birds respectively. As a rule the majority of the organisms making up the flora of the intestinal contents of these birds belonged c+ to he genera Salmonella and fiberthella. An average of 1-1 20.9 per cent belonged to tne former genus while an average of 55.5 per cent belonged to tEe genus iberthella. .p‘. Table IIO‘OZJCI' " “ “ _t . a Percentage “occurrence Of organisms in the rntest 1nal contents of’twenty hens affected with enteritis a- -- -a.-.—-. UN '- —..——-r-. “Y,- --v-r' *‘-.l -- mr‘ ”0‘ ‘O-"I. 'rwvwa‘ n.- Or :anism : Q - Bird _ _ _. - . . L- : v‘ V :3 3 It?“ ' 2 I‘— ' "'""’ 4. * P o 1 o s 2 O a o ‘ ‘ " " . ° 0 I I I o o . I O D D 0 D I a o 0 c F‘F . a . - J ‘ 9 “ 0 ° 4 : 5 z 6 2 7 e 8 z . 30 . 1] . 12 - 37 . J4 . 5 . . . . . 99 . 9 ~ w 1' ' '~ a b o I 0 I L n o a t, c A; \ ‘ -, _,:::.q._ ...-g . . . « m- p . *w-v x.- “vutm- ave-coo” - w V‘IW” ' r... NM* 0 r.:....0. nr—Iv ---—.> H 'M-homw' H “"'" O 4 J _ .. -. _, _ _ m*w' rw- H ...: 1~ - :— . ... 1 - .- :—.: 0". M --I ou-‘nrv-u—«co-e—u. n-a'r - -. I~WW-- .O-v- .---"‘WF.W ..- VW-ng-v A- n. . .o -. .1 V . t w r?*-:_':--u.: ‘3‘: :—: r : m r-‘v—vo-Iz. .- ----~-- ‘40- ~~ ’2‘“... “::. -. .o ...—.- uo- .. - q .. ~W~>A-~-~vw 'M--l ~-~' ~ «nov- . :l a . O ’ . . a a o o 9 = 9 a o c a a o o n I r Q t 3 I '— R h coli *0) 9 f e . : a? 2 2. :32. 2 '45 7 ‘ 0 l ' 4 O ' 4 6 :34. 6 * ' o - ' - 9 ' - , ° . , -‘_ ...(\“‘,.wJ . - .. ~ J Q - ., / . o a. 9 . o . a ‘ . n 9 (-4. g I I H‘nw‘p—n w-.‘ .m *- ...- ‘neo-c“ .4-1" —,. -.- «,- a" o. nww -— v ‘ " "' " ~ 7 -~ “aw--m-‘n-W H—n.~." -r-m w.-- vum ,‘.,\_'..- I"... m1-~-m-Hn—wmuwm:m I o 8 " "' : “ : a 2 . 9 O O O : I o n I I I Q B I I G O 0 G O D O O I C) . a . f. "r"! _ .. o c e 2.1 , o t 2, o . Sb . ) N l c 1n1n10r 9 4 10 . . . _ . . y . Jescn4 11~1 1 . ~ . . - o~ ~ .: '1 '- 111-: 93- . ~ 7. - - - v- ~~ o~ . ~~Jo~ - F— w w—— 9 ,,. —--n 51%“ a . Ah-w-nfi f -“ '- " ”" ’H'.“ . A" "3" 3-. " H.3- H" -’ W- 0- wmww ..o- m'ifl"- .. ‘— » v. mm:- a <0-” -——- cow war—war... . g a o a I U C o A. I I I I a o O a o . g g. a D 0 i I o I o I o I a o n 9 U ., . ' o C‘ , o 9 I a * 9 0 I o 9 5 o ‘:) f) 5 a I e o a I e 5 '7 g. I ..e‘D. C 18 o.\)l'!. . O a 'v 9 D ‘8 N ILJLQ o a I a ' o QL) n o A ... --. v.4 4‘ v.- ”90”" —~ av ‘ m‘. '0' 4"" a... a. “. I'u‘. at '00” -~.—.‘ ..- -w‘. l‘ - 3 ‘7‘ v v“ W -‘W '. . 3’- -—. -ourbl -‘ :... , o. e- -.- .. A- --. n-- -7 »- — t ‘ -- «a- .. qn-~.~—.- —-‘r-—-0-~.-r .4 ‘ ‘- . ... ...—w ......c‘ A.” , ”Lm’ “.....- . O I O O '0 .0 I I I I O I o g 8' t D ‘ O D I O D 9 t O 9 0 q. o g Q a n n a I 9 I I I I I I a a C 2 I I I g . ; L) . n a I I G I v o o n w ' _ w- - -v.‘ --. ’1'“ fl’ - ’wwm“n wm'wuaafl- -. ”'04. v-o--—‘. x. n.‘ W". .—“'~mm *“fi 0“ ~ ‘1.”- fl ‘ _ “ r— A“. - .‘V'. - ‘. -‘- o I D o I I I o a I e o a 9 p n .9 g 9 t n a O 9 v u o 9 I o p a g a . . J. 3' .1 . A . O . g a o n o n I I a I o o O c o 7 O a o 3' o S 1)» 'J .1. J. S a a. a E?) 9 I o o I a I I o I . ' ° ° 'v) C ‘J u- -5- ' ...‘-.‘ - ..- -~., ‘ _-‘__‘A-“ "__._ '- H“; "v ”Mm, rwfiq...‘ -...1' ."_'_- m .v H. mm» W q -I.. M- -- ' - - ...-.. --— ——o‘—- H— , m‘n—oowv ’n-~ mm a I I I I I I l o o a I a I o I o I I 0 O O I? D I O D a I u 4;. o u o a a g a 3 a o f) l .4 I o a o I I a a o o o o o a c . rot “Gus VI] NC‘JI ‘iS . I \J. n Irv/.1 o a I a I an a o 9 .,. a , n ‘t- r: 0 Q wv~~ .0 ... Io. -- -~ ~. to-r-I- -- 9 - nrm- no co -' ”we... .- - . .1 .r ng—u..- .m.v m‘mfi -.~- ...-w H u:- ~~M~ ’-.< _-—_. -\.-v- «I... _ -"- n---~.-—~cp- M n r—ou-‘Iu—hm n. .., ... 7- -.——.-mrau ... >W -.-WMA ~---.-rwa~ a. - ~—-" m o o n a 0 F a o o O I o o o O I I 9 o I I I a a" o I a o u I I u I o- o L 't,‘ r‘ a T7 3 ‘ a {it I 8 a 9 I I I o a e a V o a o a i 'I J r 18 a, a . Q .2 o O Q \ e a o I r o I o I o W ‘7‘."‘ww-h _. - . '._ ...... ...”..- _‘ ' .. ”u ~~~ J m “_ _- ' ‘ «a m _ ‘ ,r v _. _-——.. .- ' ow-v . .... ‘4» I." A‘— -- ‘- «...... m .- ..- -. ... ~o. .- vo—w ~04“ v. .. M ~ -- nun—4' ..-.-.r‘_‘.~»m-—I-u.- -k‘I—-~ «o—vm m a m o a o 4.- 0 O o c I I o o a e ' I I 0 ‘4 W 9 0 F ‘D ' I o a a e a .. o c I \ . Sta oh albus " “ ‘19 4 ’ ° ' ‘ 0 ‘ ° ° ' 2 ‘ ' ° '15 ' ’ 6. 8 * ' * ' 1 O '- |- s O 9 v, v/ 0 O O o a . a a s I .a. Q Isl A. n u u I n w”. an“ w- r. -.-.- -‘M~‘\.’ -.r .....— , v-v W_A*m‘~*1' ‘H “ "V’A"“~"' *W“ ‘QMW‘HN’fi‘ *“‘ """"‘ "'.".'" - ya...— u» --- 1 _ m~‘x‘-r.~"-w- "’1‘. --«~ ~ - n a.» mer cH' «c—m O I I o I I c a a o o a a a O I o o ‘ r- a a n a 9 O o w k . e a I I O I O p I r O L ‘3 1 Salmonella ~ . ~59 - 3 ~ ~50 9 - . . .57 4 . «on o . . a ~ s 4 . "56 n " “ ° .2) ‘J ' ‘ 3‘ (I O 0 a “£4 :2... s: o- . i w a 4-4 ‘4'. 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O 3 O F O O ‘- —- .. ,‘___ I o O O O O I O o n I b v Q o an A4 Q I D O 5 BF 0 o p 0 0 0 a O I O I I -. a . ,a 9 J.- ’9 o z c I! o I O D I - 93 c- 1' <- O . cereus ' 9 ' * “ c w, o g o I O n I .0 I D o n a g a I 9 O G 0 ° N D g g p I 9 D C Ir 0 Q a Q 2 U . . 9 Q g b 1..-...“ w w 0 9 C E O Q ,q 0 0 1| . . . O O I O G C O O B. me atherlum ' - . n e * ' ° - 0 . . ~ . . . . . . , : a I I O . a O O G O o o o p .p O o 5 I I ' g g I I I o O D O O I 3 I o D I O D 9 1' ——.~_ v— f. :v Salmonella‘p\ : - . . ¢ 9 ‘* . - - . - . . . - 9 . . » . - 4 . .1 ... , - -- _ I o I 6 0 I 0 o a I a a a a O o i 0 ‘ ' fl : a o I o o a I a I- e o a , n 0 .o 0 G 2? 6 " 9 C 0 0 0 D 0 O o I I a i o c u g o o - . Q c 9 _ ".———__ . w B O C Ohaer e n S g z a 0 O o o 9 I D a e I a o .9 I 9 o 4' O I 0 . O I = I O t a I a o o o D I o . . . g . O C 0 o g C O a 0 a a a g I e -o 2 O a -... g g I . 9 o . I p g 4- a o o '0 " O 9 . ‘ fl.” . . 0 O O . . O . O o g , . Q g o .0 .0 . . . O o O O 9 l D O I I a o o a 9 ‘ 100 J O - . 00 z 100 ’ lOO ' lOO ‘ ' lOO ' lOO ’ 100 ’ lOO ' lOO ' lOO ‘ 100 ° lOO ' ' O ' OO ' OO ' lOO ' lOO ' > i " I g I I 9 Ir 9 o o a 9 a _ 9 (t a .- 18 CELLULOSE DECOKPOSITIOH PROCEDbe. As cellulose decomposing organisms have been found in the feces of practically all domesticated animals an experiment was set up to study the cellulose decomposing organisms occurring in the feces of the nor- mal adult bird. Samples of fresh feces were collected from the cloaca of fifty hens in sterile test tubes. A portion of feces the size of a large pea was transfered from each sample to test tubes containing Omeliansky's medium (18). After transfering the feces to the medium one c.c. of sterile oil was placed on the surface of the medium although it was later found that the addition of this oil was un- necessary providing the tube was tightly sealed. The tubes were then incubated at 37°C overnight after which they were allowed to stand at room temperature. RESULTS. Readings were taken at seven day intervals as indicated in table No.5. The first evidence of cellulose decomposition occurred at the end of the sixth week. Additional tubes showed evidences of cellulose decomposition until the fifteenth week after which 17 tubes remained in which decomposition failed to take place. At this time the filter paper in 55 tubes was completely wilted and many of them completely decomposed. Microscopic examination of smears from the medium revealed a variety of organisms. However, in most of the Table No.5. CELLULOSE DECOKPOSITIOH J Time 5 Condition of filter paper :1~:.. ‘* : No Ehange : SIightIy wiltedft‘CbmpIeter : Q : wilted :No. of tubes : No. of tubes : No. of tubes 7 days Q 50 Q :1 14 " Q so Q Q 21 " Q 50 Q fi_ Q 2e» " Q 50 Q Q __ 55 " Q 50 Q Q 42 " Q 48 Q 2 Q 49 .. 4o a w 2 56 7 Q 31 Q 15 Q 4 63 7 Q 27 Q_ a Q 15 7o " Q 24 Q 5 Q 21 77 " Q 20 Q 7 Q 25 91 " Q 17 Q 5 Q 28 98 " Q 17 Q 2 Q 51 105 7 Q 17 Q 0 Q 33 _112 " Q 17 Q 0 Q 33 19 tubes gram negative rods and gram positive cocci prevailed. The cocci always seemed to be in closer contact with the decomposing filter paper while the rods seemed in great- er abundance in suspension in the medium. In many tubes involution forms, while not in abundance, were consider- ably in evidence. Spores were numerous in many tubes, while Actinomyees were abundant in a few. All attempts to isolate the organisms responsible for the decomposition of the cellulose in pure culture failed. While many of the organims of the fecal flora are not capable of growth in this medium, they are no doubt able to survive. It seems that more than one species of organism or symbiotic relationship of organisms was responsible for the decomposition of the cellulose as the filter paper in all of the tubes was apparently not attacked in the same manner. In some tubes, translucent spots sur- rounded by a yellowish brown ring appeared, in others, the filter paper became fringed at the edges, while in others, the surface of the paper became fuzzy. In most of the tubes the sediment tended to become black or slate gray probably due to the fact that hydrogen sul- phide was formed in most of the tabes. It is interesting to note that repeated transfer of small pieces of filter paper or at least 1 c.c. of. the liquid portion of the medium after the filter paper began to wilt greatly shortened the period in which the filter paper was decomposed. In several cases after four transfers the period was shortened from eight weeks 90 originally required for cellulose decomposition to three days. The transfer of very small pieces of filter paper or of very small quantities of the liquid portion of the medium failed in each instance to produce growth. If small pieces of filter paper were smeared on nutrient agar and salt free nutrient agar abundant growth occurred during twenty four hours of incubation at 37°C. If this growth was heavily transfered back to Omeliansky's medium cellulose decomposition again be- came very slow and required almost as long for decompos- ition as originally. This was true even though the cul- tures were transfered after twelve hours. If these cul- tures were transfered after 50 to 36 hours cellulose de- composition failed to take place. Transfers made to nutrient broth containing filter paper and incubated at room temperature failed to produce cellulose decomposition. 21 VIII. pH OF THE FECES OF EORHAL ADUIE BIRDS. PROCEDURE. Samples of feces were collected from the cloaca of normal adult birds in sterile test tubes. The sample was thoroughly emulsified in neutral sterile dis- tilled water. The pH of the sample was then determined by Youden's hydrogen-ion concentration apparatus. RESULTS. 1. 7.115 28. 8.040 55. 6.690 2. 7.285 29. 6.945 56. 7.615 5. 8.210 50. 6.860 57. 7.570 4. 6.775 51. 6.550 58. '7.570 5. 7.450 52. 6.945 59. 6.455 6. 7.785 55. 6.265 60. 5.585 7. 6.095 54. 6.860 61. 6.180 8. 6.455 55. 7.200 62. 6.550 9. 6.265 56. 7.200 65. 6.945 10. 6.455 57. 6.690 64. 7.115 11. 7.700 58. 6.265 65. 7.115 12. 6.945 59. 6.550 66. 5.840 15. 7.615 40. 7.550 67. 6.690 14. 7.050 41. 6.945 68. 6.860 15. 7.550 42. 6.5204 69. 5.925 16. 6.265 45. 6.520 70. 6.860 17. 6.860 44. 6.455 71. 7.285 18. 6.690 45. 6.605 72. 6.945 19. 6.180 46. 6.180 75. 6.550 20. 7.550 47. 5.500 74. 5.585 21. 7.115 48. 5.250 75. 7.200 22. 6.605 49. 6.455 76. 6.945 25. 8.210 50. 7.285 77. 5.500 24. 8.210 51. 6.945 78. 5.500 25. 6.520 52. 7.200 79. 6.095 26. 6.860 55. 6.690 80. 6.520 27. 6.690 54. 6.945 81. 6.455 The pH of the above samples of feces varied from 5.5 to 8.2, the average of the eighty one samples being 6.72 . Q ‘3’ 1.5.0 ..V‘ IX. DISCUSSION. Fundamentally there seems to be very little differ- ence in the bacterial flora of the feces of the ten two uhekkeld normal chicks and the twenty normal adult birds studied. However, there seems to be more stability in the occurrence of M. .921; and 2.5.9.30 gommunior in the feces of the adult birdthhan in the feces of the chicks. One or the other of these two organisms was absent in the feces of three chicks. Thirty four organisms were found in the feces of the ten chicks while the same number was found in the feces of the twenty hens studied. In health it is probable that there is a tendency in early life to acquire a flora in which $2220.2911 and Esch. com- munior are to predominate. It seems very apparent that Eggh.‘ggli and Eggh. communior are the predominating organisms in the bacter- ial flora of the feces of the normal adult bird as well as being constant inhabitants. The remaining organisms apparently are of little significance and their presence 'is no doubt governed by certain symbiotic relations that exist from.time to time or by chance. Even birds under similar conditions of environment have a wide range of variance in the species of organisms present, excepting of course the colon organisms. Sal. pullorum was found in the intestinal contents of all of the chicks affected with pullorum disease. 25 The fact that this organism was found in the intestin- al contents of two chicks even though it was not iso- lated from the organs of these birds would lead one to believe, as suggested by Mallmann (17), that the intestin- al contents as well as organs should be cultured when autopsies are held on suspicious chicks. ‘§§l.'pullorum was found to be present in the intestinal contents of one diseased adult bird to the extent of 26.4 per cent. Thus it would appear that adult birds may in some in- stances spread this organism in their droppings. It is not known whether this bird was a reactor to the agglutination test for pullorum disease. Organisms found in the intestinal contents of the twenty diseased birds consisted chiefly of organisms of the paratyphoid and typhoid group. It seems very likely that the presence of worms or coccidia or both in the intestinal tract may produce a condition in which the percentage occurrence of Eggh. gal; andggggggggg- munior is greatly reduced thus allowing the establish- ment and multiplication of pathogenic organisms. The marked presence of paratyphoid and typhoid organisms in the intestinal contents of the diseased adult birds would lead one to the conclusion that the colon type of or- ganism does much to prevent invasion of the intestinal tract by pathogenic bacteria. It would appear that possibly some of the acute deaths occurring in coccidiosis and worm infestation may be due to inva81on by pathogenic organisms through the inflamed or broken mucosa and in a few instances may even be due to invasion by the colon organisms them- selveSo It is very unlikely that cellulose is decomposed in the intestinal tract of the normal fowl. From the experimental data it would appear that cellulose is not decomposed with any degree of rapidity unless great numbers of cellulose decomposing organisms are present. Since the time required for food to pass through the in- testinal tract of the bird is comparatively short it would appear that this time is much too short for cell- ulose decomposition to take place. It appears that the colon organisms are constant inhabitants of the intestin- al tract of normal adult birds and since the intestinal contents are a more favorable medium for the growth of these organisms it would seem that they would tend to in- hibit the growth of cellulose decomposing organisms. While it is true that the feces of some birds are semi- fluid in consistency and contain practically no cellulose no accurate estimate can be made of the amount of cell- ulose individual birds actually ingest. “It is no doubt true that many organisms that invade the intestinal tract of birds cahnot be isolated by the procedure used in this study. These organisms are prob- ably of little concern,however, as far as disease in the fowl is concerned. The average pH of eighty one samples of feces taken from the same number of normal adult birds was 6.72 l. 4. 5. 25 X. CC “I"SIC'TC . Organisms of the colon group appear to be constant in- habitants of the intestinal tract of mature birds. They represe.ted an average of 60 per cent of the total organisms present in tne feces of the twen- that 01 the adult birds except that there was more 0" less instability as far as the presence of or- In this study all of t1e ten c1iclcs affected with pullorum di ise ese har‘t ored Sal. pullorum 1n their intestinal contents. Twenty adult birds affected with enteritis showed a pr edominence of organisms of tlie paratyphoid and tyghoid group in the intestinal contents and a very narked decrease in the number of colon organisms present. The findirg of Sal. pullorum in the intestinal contents of one hen w011ld lead one to believe that some adult C11 2170‘)“ —L._ birds may carry and spread this organism in their Ho 3 02 U1 . 0"" "D “I'm .F' ..H_ -..V' .l 0 '0 ~5‘ :] .... r. ‘ 1' N n a. L _1, : ‘ r . -::.‘L.~I'lgj 34-3.14; ' k. A 11;: l. I‘ 11'.Aj . 0., ' .’M. 324.21 DD 03 —m_--'”i-_w.'~“r " M J‘w; 4 ‘4 . i. 6. A large number of samples of feces of adult birds decomposed cellulose when placed in Omeliansky's nedium.but it is doubtful if cellulose decomposition 1‘_ occurs in the intestinal tract of birds. 7. The average pH of the feces of 81 healthy adult birds was 6.72 . ... -' '1 _ srwrurr .1 _ . '. 7 —=r~—* E‘:‘-" -4!- , wu‘ r‘_‘ (l) (2) (3) (4) (5) (6) (7) (8) (9) (10) (ll) 27 XI. BIBLIOGRAPH‘. Kern. Beitrag zur Kenntniss der in Darme und magen der Vogel vorkommenden Bakterien--Arbeiten aus dem Bakteriologischen Institut der Techinschen Hochschule zu Karlsruhe. Vol.1, (1897), p377. Schottelius. Die Bedeutung der Darmbakterien far die Ernghrung. Arch. f. Hygiene Vol.34,(1899),p210. Schottelius. Die Bedeutung der Darmbakterien far die Ernflhrung. II. Arch.f.Hygiene Vol.42,(1902),p48. Schottelius. Die Bedeutung der Darmbakterien far die Ernahrung.III. Arch.f.Hygiene‘Vol.67,(l908),pl77. King. The Bacterial Flora of the Intestinal Eucosa and Conjunctiva of the Hormal Chicken. Jour. Amer. Med. Vol.lO,(1905), p400. Gage. Bacteria and Animal Organisms found in the Feces and Intestinal Hucosa of Healthy Chickens. Bul. 155,(1911).~Haryland Agric. Exp. Station. Rahner. Bakteriologische Hitteilungen fiber die Darm- bakterion der Hfihner. Zbl. Bakter. Vol.30, 1901, p 259. Henes and Rochlin. Darmmikroflora des Hausgeflflgels. Zbl.f.Bakter,Orig. Vol.115,(1929), p521. Burke. Burke's Modification of the Gram Stain. Jour. Bact. Vol.7, (1922), p178. Bergey. Manual of Determinative Bacteridlogy. (1925) Weldin. The colon-typhoid group of bacteria and re- lated forms. Jour. Science (Iowa State College) Vol.1, (l927),2, p121. (12) (13) (14) (15) (16) Ford. Textbook of Bacteriology. (1927) Voges and Proskauer. Beitrag zur Ernahrungs phy- siologie und zur differential diagnose der Bakterien der hamorrhagishen septicamie. Ztschr.f.Hygiene.Vol.28, (1898), p20. Clark and Lubs. The differentiation of bacteria of the colon-aerogenes family by the use of in- dicators. Jour. Inf. Dis. Vol.17,(l915),p160. Levine. The correlation of the Voges-Proskauer and ‘Methyl Red reaction in the colon-aerogenes group '“ of bacteria. Jour.Inf.Dis. Vol.18,(l916), p558. Clark. Differentiation of bacteria of the colon- aerogenes family. Jour. Amer. Water Wks. Assoc. (1918), p26. (17) hallmann. Salmonella pullorum in the intestinal con- (18) tents of baby chicks. Jour. Inf. Dis. Vol.44, (1929).l. D16. Giltner. Laboratory Kanual of General Eiorobiology. (1921) 29 ACEIOWIJGDGEIEJHT . The writer wishes to acknowledge his indebtedness to Dr. H. J. Stafseth and others for their assistance and helpful suggestions during the course of these studies. . .-. . 1 . . . .i: - ,I , i . . .... .11: :1 -_ 2.41... “tithing 1 I. .3“ 33.1 1 s. . 8. 23.147... .5 .. ...,wfluanwsnfiflflnum‘ hm...) 35339.4. .I (I! ll 1 ‘fnyx. I . . x 1 .,.VJ(‘|N,Y l. :1 billbvvl 9?” Raw ,. (€1.81ka LOAN USE UM? M1? 14 1192; Fr MY 0 '53“) .r . .... . ._ ._ . I n . {I - .... . ...)...Jfiu u. \«r.. 9- .43.. .111 a. 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