HI I 1 Hill“ I I 145 964 HTHS . 4“ A 3". \‘ A H k “ rs_¢.¢.-. - 7" I; : Tl—IESIS A STUDY OF SOME BACTERIA ESOLATED FROM RETTED FLAX FIBER Antoinette Trevithick I 924 3129A 3263____ » x MIMI: mmmanmnnw ., A , ,fi.—_—_ PLACE IN RETURN BOX to move this checkout from your record. TO AVOID FINES rotum on or before date duo. ‘ DATE DUE DATE DUE DATE DUE msu luAn Afflrmativo mason/Emu Opportunity Institution W pus-9.1 V i —_....__—————_ I llfuillir »;fr.faln1v.»5.. .3: :1 f r; ‘3‘:th . iv. ‘ , Arr-EAEDIILL'wLLKK ., A STUDY OF SOME BAGTEIHA ISOLATED FRO]! RETTED rm FIBER A 31331 0313333 33333413 IDUlaTBU .1. .'2 .‘TI’ l-J - I .-‘ ‘. ‘, ARLM jjflluu Slug 2 JJR Theei 02 Submitted to the Easpfity of t:e hichiran A: \1 L; U“ ricultural College in pavtial fulfillment of the requirements for the degree of has er of Science. By Antoinette mtevithiok 1324. THES:S I3, U." ‘7- f\ g-. *1 "“11. ; ‘;'~,' ' "" .LJJJI 2J1 U'\:.‘:...J -:&L.I‘.L'_J-Ll.’l IQVquIJJ‘AJ EiiCIJ -CSI'CLIBD ELAX 1"]. BER. fl table or Jonten s I Introduction 11 leview of Literature III methods of Investigation and Jesults 1. Isolation of urgani sms and oouroe 3. Jaltural Studies on Standard Ledia a. Jellulose and :eotin Tests 4. Oultural Studies on Synthetic Kedia (a) Usehinsky's ASparagin Media (“1th and Without Ieutin) 1b) Jonn's Solution (with end Jithout reetin) (o) ainOgradsky's hedia (with and Jithout rectin) (d) “inOgradsky's ASparaginate Agar (Jith and aithout Ieotin) (e) JinOgradsky's agar for Aotinomyoetes (With and ”ithout reotin) (f) carrot Agar IV Discussion of Aesults V inmrmry VI BibliOgraphy 1'. ‘x ‘1“: ..... fli::") I - Iii 1’10 DU.) 1'1 i211 Jotton fiber, which is superior to flax fiber in purity, has frequently been the subject of investigations the purpose of which has been to date mine the factors causing disintegration in cases of wet storage. huch.more favorable are the conditions for microbial growth in scutched and heckled flax fiber. rhe reason for this lies partly in the chemical composition and rartly in the different anatomical structure of the two fibers. Bast fiber, which is found in flax, has no protective cuticle, and in the central lamellae there are mineral substances, carbohydrates, fatty and waxy substances.(l). Due to the biological process of decomposition by which fiber is retted there is naturally a large variety of organisms present on the surface of the fiber. Such.organisms as Bacillus mesentericus, B. subtilis, B. comesii, Jlostridium anylobaeter, Granlobacter Qectinovorum, g. aerogenes and many molds and yeasts have been pointed out as the chief causal agents in flax netting. At least it is definitely considered that rettin? Since a flax fiber is made up of individual cells held together by pectic substances which are exceedingly hygrosconic (15, 14), the purest fiber will always maintain its great water-attracting power. Even with moderate moisten- ing of scutehed fiber which contains such an abundance of food material, it seems reasonable to believe that many to I organisms deveIOp on the fiber. Under storage conditions, aerobic organisms probably deveIOp. This project was undertaken primarily with a view to determine the type of aerobic bacterial flora ey— isting on the scutched flax fiber. II - hIoIUJIJAL JBIISH LE LIQJAAL'JJJ fery little hork has b an done with the organisms present on rotted flazr fiber; more work has been done from the flax rotting standpoint. One would naturally xeec- some or J. the Organisms which are present in the rotting solution to [.3 be found on the retted fibe r. f. In 1885 uincgradsky (6) found an.aerohic spO"s~ J.“ no 9 . . .",. ‘T. . ,, N. ‘A 1 r ‘ , - ~ \ , - ‘0 iorner Nulii as cesi rated as the spec lie or: anism causing retting. In 1902 fa“mann (18) stated that many microorganisms, t C evicus, B. subtilis, streptothriz sj., such as J. meson _ Bseudomonas fl uores ens could set flan. h Wijei ick and VanDelden (1905) also found another organism, Granulobacter pectinovorum. jossi (1916)(lf) described the specific organism, Tanner (19.3)(5) found that the flora from retting vats varied, but Spore-forming bacteria of both aerobic and anaerobic tyres mere common. whe aerobic types were similar to the members of the subtilis-mesentcricus group; they formed large spreading colonies on solid media ar’ liquefied gelatin very rapidly. as found the anaerobe clostridium p...) C? amylobacter to be the most Specific in retti1 this organism as follows; (1) Vegetative Jells,grown on common media, large rods with dense protoPlasm. (a) shores, larger than vegetatiVe cells, clostridium shape. (5) fermentation deaction, large amounts Of gas formed in lactose, glucose, saccharose and glycerol. (4) Litmus milk, curd peptonized. (5) Gelatin, quickly liquefied at 20° 3. (6 Plain broth, rendered cloudy. (7) Nitrates, reduced with the formation of nitrites and arm on is . Auschmann (l) studief the organisms on the fiber of the heckled flax by staining them on the fiber. He used a process that had as its basis the ordinary Gram stain. He found that when the fiber was re ted by the steeping method the predominating organisms were dram positive, for example, 3. mesentericus, B. Asterosyorus,_§.‘megatherium, g. subtilis, the cocci and streptococci. By his Gram coloring method he found cocci,a "tender" narrow bacillus, small oval bacillus, rebust long bacillus, Spores, yeast-like organisms. he could not find b. an;lobaeter on the fiber. He recognized the fact that artificial drying was able to kill a number of living germs. as though the majority of coccus types were due to air contamination. He also worked with liting organisms on the fiber and found that the aerobic organisms occurring most D frecu ently on the heckled flexes, uater-retted and artificially dried,were 2. mesentericus and B. megatherium. lhe first would include B. mesentericus fuseus, h. mesentericus vul satus, _ m4 related or varieties. rhese, with 2° ieaatherium were the only aerobic spore-bearers found. he found that the anaerobic retting stimulator (clostridium anylobacter)adhered to the fiber in a living ferm in Spite of artificial drying, but he did not find it on the fiber when he .p3n lied the modified Gram stain. the vegetative forms of organisms were more numerous on the fiber of a naturally dried, dew-retted flax. while the aerobic Organisms predominate on this, the anaerobic and "potentially" anaerobic organisms were more numerous on the fiber of the artificially dried water-rotted flax. host of the orgaxn sms were capable of ecomposing pectic substances. he also stated that cellulose consumers are usually found. Makrinov (ll) referred to a pectin-fermenting o-ganism isolated from soil. he named this organism Iectina- bacter amquBhildm. This is a motile, aerobic rod, measuring 4 to 6 microns in length by 5 microns in diameter. uuring eporulation.the microbe takes on a fusiform aspect and in the enlarged end of the rod an ellipsoidal spore is formed. fhe organism is Gram positive. In bouillon a slight turbidity is found and at the surface, large lamellae are present. Milk is coagulated On boiled potato, vise id grey—1vhite col nie are formed With a strong production of gas. the ontimum temperature is 30°to 35°c. fermentation is most energetic on fresh potato, less energetic on cooked potato where the starch is partly hydrolized, and feeble on beets, carrots and turnips,uhich have sugar instead of starch. He decided that this organism was an active agent in pectin fermentation (in flax rotting) under aerobic condi- tions. 'ccording to Sustaf Sellergren (5), Antonio Bozzocchi (3), Thatcher (14) and others, the fiber bundles are composed of cellulose and the eel s are held together with a substance called pectin. The“e has been other work on flax retting and pectin fermentation, but only of a general nature. ‘ ‘1 11: (\1 \‘r,'\ w.: y -. -~ x,‘::?“( K. III bLJAALV \,a ll. JJQ-LLJ‘LJ. Cali 5..le -iA'JUU‘dAIQ 1. Isolation of Organisms a2cd Source. during the summer of 921 in connection with some :J expezim work on flaI: re “tin, a large number of organisns O was isolated from water-retted f ax fi3e r. Chis fiber had been scutched, that is, after having been through the breal :ers, the shives were re moved from the fiber by means of rotating heaters. Shilean, Dutch, Irish, Jourt1ai,hichigan and Japanese retted (5 H (D fibers were used. Bron these fibers aerobic dilution plates 3 o :3 “ade in the following way. A small piece of fiber was taken f the center of the fiber bundle and placed in a flask of normal salt solution, allowed to stand for a short time, and then shaken well. from these flasks ordinary dilution plates were made, using nutrient agar, pH 5.8. Lhes e rlates were incl 48 hours at 25° 3. and 30° J. leprese ntaciv-e t"“es of colonies were picked from these plates, transferred to agar slants, grown 84 hours at 55° 3. from the 24 hour growth iram stains were made. If the culture as were not ours they were replated until pure, as shown by microsconical examination. 33 this nethed 91 cultures were isolated. 2. Jultural studies on standard hedia. .3 ng, the e17tures were grown on the various H. H mm. - v. . alter puri J H- ,4. ‘d l kinds of media used n identification. for this work the (escr tive charts re ”c01;nende d bgr the society or aneriCan BacterioIOgis -8- II!!! and. Ear methods of medic-making the directions given by the “Mcty ofihmcricnn Bacteriologlctc and Giltncr'a Hanna). were hum. t‘or methods at making nutrient user (using 3 no. but Ctrcct per liter). gelatin. glycerin potato, nutrient broth (using 3 me but extract per liter). dextrose, lactose. saccha- rose and glycerin broth am starch agar the made): in ntcrred to those two manuals. All or the motto were adjusted to pH 6.8, and bran maul purple can used an indicator in the sugar hrotha. In.cll cacao the cultures were cot up four different time. 1n duplicate. and In all characteristics ca glvcn.thcy checked. Yer the last two dotennlnntlons a now not ot’mcdlu was made. unincculatod controls were run with each aXpordnmnt and all tubes cox-c incubated at mm. wlth tho acaptlm ct gelatin platen an! tubcc which can incubated at com. gcgultg. After purification.thc culturcc were given stock numbers as fellows: Stock no. source of Isolation stock ho. Source of Isolation z chum 111m: . fl chum fiber 4 I! I 23 N I! 5 '0 n 86 n n 6 " ' 31 Dutch fiber 10 " " 33 Irish " 13 n It 34 It I! 13 " w 35 " n 17 n n 37 n n 19 n u 38 n w stock MD. Source of Isolation steel: 30. source of ISO lotion 40 Irish fiber 76 Japanese fiber 4LZL z! I! fi7f3 _ I! :1 46 Courtrai fiber 82 " H 68 Iiichi gnu " 83 n n 69 " " 85 I! u 72 " . " 9O hichigan fiber 74 Japanese fiber 91 Goururai fiber rhose numbers uiich are missing in the numerical sequence of the nrevious table belong to orgeLisrs which are dueliectes of some of those described. .A Ehe organ is me in the followinq descriptive charts have been grouped as fellozs: (a). Groun I.- These rod-shaped organisms which have eccentric Spar es which ore not enlarged on slaorulstion. (b). Group II. ihose rod-shaped organisms which have eccentric spores trod \Wiicfll INN) club—shapeion eporuletion. (c). eroup III. whose rod-shaped org-nnisms which have central Spores and uhich are sgoindle- slwao d on Sporuletion. (1). GrOupQLI. small rod-shaped organisms which have )ores and “lich are not swollen on eporulation. (e). Group V. Jed—shaped orgunisns without Spores. (f). Group VI. 302:3 us forms. 5—.” - 1L - UXGAJISJB STUDIEJ Groun I. Organism HO. 5. I . L10 rphol 03;; 1. (‘9 00 4. legetative cells: (3) Born, medium rods, round ends. (b) Limits of size, 3.5 x 1 mLmon.- 5.5 x 1.5 nicrons. 25nd DeporeS: (a) Position, slightly eccentric (b) Form, elliptical (0) 30d not swollen on Sporulation motility, true staining, Gram positive II. Jultural Sharacteristics: l. Agar stroke (a) Growth, abundant (b) Form of growth, echinulate (3) Elevation of grwoth, flat (d) Luster, dull (e) EOpOgraphy, contoured (f) Optical characteristics Opaque &. Gelatin stab. (a) Growth best at ton (b) Line of puncture, filiform (c) Liquefaction, saccate to stratif rm 3. Potato media (a) Growth abundant \ (b) Eorm of growth, Spreading (c) Luster, dull (e) Torograrhy, contoured, wrinkled (f) JhromOgenesis (g) Betato, dzrhene 4. Nutrient broth (a) surface growth, none (b) Cloudiuv U, moderate (c) Sediment, viscid 5. Agar colonies (a) Growth, rapid (b) Form, circular; sub-surface, irregular (d) Alevation, flat (0) Edge, undulate 6. Gelatin colonies (a) Growth, rapid (b) Elevation, flat (O) Edge, lobate (d) Liquefaction, complete in a dag III. PhysiolOgical Features 1. fermentation tubes, brom cresol purple plus Gas , none (3) Destrose deaction, acid Gas, none (b) Lactose AAA AAA Reaction, neutral (c) saccharose reaction, neutral ( Gas, none (d) Glycerin ( ( Reaction, acid 2. Litmus hilk, peptonization in 2 o. Ammonia test,positive in 10 days 4. Indol, positive ‘1‘. “J8 (nitrate pentone) 5. Diastatic .ction, positive in 4 days 6. Temperature relations (3) Grows at room temperature (b) Grows best at 50° 3. (c) Grows at 45° C. Organism Ho. 41. This organism compares well with NO. 5 except as follows: I. morphology 1. Vegetative cells (3) Same (b) Size, 2 x 1 micron - o x 11. Jultural dharacterisitcs 1. Agar stroke (a) Form of Growth, Spreading (b) Luster, glistening to dull lII.PhysiolOgical characteristinq 1. Fermentation, same except ( Gas, none Glycerin ( ( deaction, neutral 1 I111 Bron. Organism NO. 69. Same as ho. d1 except II. cultural Sharacteristi C) s l. Agar colonies (a) Eorm of growth, surface—irregular. III. Physioloqical Sharacteristi L3 8 1. Fermentation ( Gas, none ( Jeaetion, acid ~rganism LO.10. I. MorthIOgy 1. Vegetative cells (a) Form, medium rods, round ends (b) size, 8.5 x 1 micron - 5 X 1.5 microns 2. EndOSpores (a) Position, about central (b) host rods not swollen 5. motility, true 4. Staining, gram stain positiVe II. Jultural Sharacteristics 1. Agar stroke (a) Grow h, moderate (b) Norm growth, filiform (c) Jlevatien, flat (d) Luster, glistening (9) Optical cha acteristics, Opaque r (f) Jhromogenesis 3. lelatin stab (a) Growth, uniform (c) Linuefaotion, saccate 3. Potato (a) Roderate (b) Spreading (c) dolor, tan (d) Smooth (e) shiny e. Nutrient broth (a) surface, ring (b) Jlouding, slight (c) sediment, none 5. Agar ccflonies (a) .}ro=.'.~'th , rapid (b) Form, surface,circular; subsurface, irre (a) (d) 1“ wtl (f. H; ace, smooth Po lev on, raised '6 L—_. m (e) Jdge, entire . Gelatin colonies (:3 (a) lronth, rapid (b) EOrm, punctiform (c) Jlevation, flat (d) Jdge, entire (e) eligrt liquefaction. III. PhysiolOgical characteristics 1. Fermentation tubes, brom cresol purple plus ( (a) Dextrose ( ( ( ( Gas, none (b) Lactose ( ( lies ction, acid ( Gas, none (c) saccharose ( ( Reaction, acid (d) Glycerin ( ( 2. Litmus milk (a) b113huuy 001d first day (b) Smooth curd second day 5. Ammonia test, positive (nitrate pcptone) i. Indol tee ,prsitive o. Diastatic action, positive 6. ”a emperat ure relations (a) Grows at room temperature (b) Grows best at 50° 3. (3) Grows at 45° 3. Organism no. 5d. 1. Vegetative cells (a) Jerm, medium rods, round end, filaments ‘1 H0 or on (b) Limits of size, x l mic;on — 5 x u 3. Jndos spo res (a) location, slightly eccentric (b) Size, fills most of cell (3)1” :ot swollen (33 O r—J p. u 0 Cl- ility, none 4. dtaininr Gram positive J II. Jultnrnl Jharsct3ristics l. “gar stroke (a) Growth abundant (h) EOrL echinalate 9 (c) filevstion, flat (d) Lister A I I; (J b H O :3 O \J I C. :5 (D L) H. U) a L4 “5 ‘J "J 3 3. fotsto (a) inset-h , a s t ulsnt (h) Jolor, fade: rel—bnogn (3) Lester, dill (1) Growth, like sewn on rotate i. hltrient b:ot3 (a) s1:£;zo grixth, : n3 (b Jloacing, clear (3) ocoirmntt, flakg :3. “3:2: cOlOJ’lic‘ ('1! J- vu'th, 3"qu (b) £03m, sazfacc, cisculnr; Lihsnrjsce, irre;.,“nell (314310W1J‘f, -, -7. {1, uddc, undulate 5, 'el“ii1 colonies (u ) juJCI.L., -rj‘l l 4. - at, 5‘ t -‘1 t- - 0 1 .V .1. , c:L*-e 3 in a ufiga A C)‘ i... £3 (J cf 0 (l (0 AAA w .l. ., :3. him-as milk ( (a) rcptonizcd in a dxgs , positive (nitrate pepton) /' “ ,. , L. .L ' ‘_ "o e. UldStcle action cosi L1 0 Temperat"re relations (a) Grows at reow tenpereturc (h) irons l‘est at ”0° d. 6. Indol, negative Organism NO. 68 I. Lorph0103y l. Vegetative cells (a) Form, medium rods, ends round (b) limits of size, 2 x 1 micron - 2. Endospores (a) yosition, eccentric (b) form, elongated or elliptical (c) Aods not swollen 3. hotilitg, true Li. J II. Jnliirnl )harneteristics 1. gar stroke (a) drowth moderate (h) Jorm, Eiliform (c) SleVstion, flat (d) Luster, glistening (G) TOnOgrnnhy smcoth J. J 9 ‘elatin stab gt, saccatc to stratiform Botato (a, Jolor, white (‘0) Shiny to dull (3) Potato br'wn (d) Ionography, smuoth (e) Anised (f) Potato, not chen 0(1 .JV Nutrient broth, slight growth Agar colonies (a) Growth, slow (h) form, circular (3) Surface, (d) Alevation, flat (e) Edge, entire (f) Shiny, white Gelatin colonies (a) Small (b) complete liniefactic III. PhysiolOgical Jharec aristics l. Ecrmentatien tubes, brom cresol nurple plus ’1 18.5 110116 ‘ 9 (a) nertrose AAA deaction, acid ( eas, none (h) Lactose ( ( Reaction, acid Ja s , none ( (e) daccidrose ( ( deactien, acid '1- ans, none (d) dljcerin ( ( Jeection, neutral 3. Litmus milk, reduction in 10 days 3. Ammonia test, negative (nitrate peptone) 4. Diastatic reaction, negative 0. Temperature relations (3) draws at room temrerature (b) drows best at 30° 3, GXUU? 11 tv ()4 0 sm N0. ; p) H. orgm‘ I Morph0103y l..Tegetutive cell (a) it‘orm medium rods round ends 9 9 (b) limits of size 2.5 3.5 m'crons - a X .5 micr01 ’ 3. Endosnores (8) Position, eccertric (b) dods, slifihtlz club-shaped. 5. motility, true 4. Staining, Gram stain positive II. Jultiral Jharac-eristics l. Agar stroke (a) Growth, scanty (b) Horn of growth, echinulate (c) Slevation of growth, flat (u) EOpography, contoured to rugose (8) Optical characteristic, Opaque (f) dhrom genesis, White 3. Gelatin stab (a) ironth uniform (b) Liquefaction crateriform to stratiform . 0. Potato (a) Growth, moderate (b) Perm of growth, chalky or lichen-line (c) raster dull (d) Toy'graphy, wrinkled (e) rotato, grey (f) Jolor of growth, grew to tan 4. hutrient broth (a) Surface growth, pellicle (b) Jlouding, Sll3ht (c) Sediment, viscid 5. Agar colonies (a) Form of growth, irregular (b) Surface, smooth (c) Elevation, flat ,3 "\ , (u) Adfic , miTulate _ 31 - 6. Seletin colonies (a) drowth, slow (b) A'Orm, circular (c) jlevatien, flat (d) Jdge, entire (e) Liquefaction, couplete in 4 days III. Physiolo;i cal Sharactoristics l. Eerznentation tubes, brom cresol purple plus (a , none (a) hext 3( ( Reaction, acid Gas none 9 (b) Lactose AAA Reaction, neutral ( ( Jas, none (c) Saccharose ( ( deaction, acid ( Gas, none (d) glycerin ( ( Reaction, neutral 2. Litmus milk, nertonizeu J o. Ammonia tests, positive in lo days (nitrate pertone) 4. Indol, positive 5. Diastatic action, positive in 4 days 6. Tempe re ture relati on (.0 (a) erovs at room temverature (b) urOWS b35313 51' 50° 3. (c) Grows at 45° 3. - »- ° . M 9) Organism no. a I. I.l0r}:h.OlOgJ . l. Vegetative cells (a) rorm medium rods, round ends 3.3 x 1 micron - 2.2 x .05 micron .‘ (b) form elliptic‘l 4. staining, Gram stain positive II. Jiltural Jharactcris 188. 1. Agar stroke (a) drowth, moderate (b) Eorm, filiform to spreading (c) filevation, flat (1) Luster, glistening (e) EOpOEraphy, smooth (f) 3hr0m03enesis, cream L. Gelatin stab (a) drouth, uxiform (‘0) Line of ounc‘ure, filiform (c) Liquefaction, saccate 5. Potato, slight 4. Nutrient broth (a) Surface growth, none (b) clouding, slifi) (c) Sediment o. agar colonies f. o. Gelatin colonies (a) Small (b) Slight liouefaction III. Physical Jharacteristics l. fermentation tube brom cresol purple plus Gas, none (3) Dextrose acid Rea-3 ti on , Gas none 9 AAA AAA deaction, acid U) ca , none (c) Saccharose ( ( Reaction, acid ( Gas, none (d) Glycerin ( ( Reaction, acid 2. Litmus milk (a) doagulation,in 7 days (b) deduction in 7 days 5. Indol, positive 4. Ammonia test, positive (nitrate pertone) 5. Iemperature relations (a) Grows at room teuierature (b) Grows best at 30° 3. - DD — Crganism Lo. 40 I. Morphology l. Vegetative cell (a) Form, Slender rods (b) Size, 2 x .5 microns 2. EndOSpore, no spore 3. hotility, slight 4. Staining, Gram negative II. Jultural Sharactcristics - l. Agar stroke (a) Growth, scan 3 (b) form of growth, filiform (d1 Lu ter glistening (0 (e) 311.1'3110301'10sis, light yellow 2. Gelatin stab 1 (a) GrOWtfl, uniform (b) Line of puncture, filiform (c) Linuefaction, none 3. Potato (3) Growth, moderate, thin (b) dolor, lemon-yellow (c) TOEOgraphy, soooth (d) Glistening 4. Nutrient broth, slight growth 5. Agar colonies, small 6. Gelatin colonies (a) Small (b) Liouefaction, slight if any in 1 month - «)1; - III. lrsiolo 1 al Jracacteiisuic 1. fermentation tubes, b; om crc sol purple plus ( Gas, none (a) next rose ( ( Jeaction, acid ( Gas, none (b) nactose ( ( deaction, acid. (G as, none (0) Baccharose ( ( Jie ction, acifi ( Gas, none (d) Glycerin ( ( deaction, acid 2. Litrn1s milk, reduction in 7 days 5. Has test, positive in 4 uays 4. Ammonia test, positive in 4 ‘a7s 5. Remperature relations (a) Grows at room temperature (b) G; ous bes t at 30° C. GiUJB VI Crganism Ho. 82 I. MorpholOSJ l. le3etr tive cells form, cocci, small 3. Motility, none 3. staining, Grsm no gative II. cultural Jharacteristics 1. Agar stroke (a) Growth, moderate (b) form of gr wth, filiform (e) (d) ulBVh ti “.13 , flat Luster, glistening 7'? o. Potato 4. Agar colonies (a) Jrouth, moderate (b) -orm circular '2‘) 9 (c) surface, smooth (d) Elevation, flat (3) TOpOgraphy, smooth (fl) Cptical characteristics, Opaoue (g) JhromOgenesis, white 5. Gelatin stab ( (b) Line of nuncture, filiform £1) ) Growth, best at top (c) Liouefaction, none 0. Potato slight if any gronth V. Afar colonies (a) Growth, moderate (b) Form, circular (3) surface, smooth (c1) ijdge, entire 8. Gelatin colonies (a) Growth, moderate (b) Small (a) Partial linuefaction in v dags complete in 1 month 9 1. Hermentation tubes, trom crosol purple r as ( (c) saccharose ( ( das, none .-— 77‘: deaction, acid (a) Acid in 7 days (b) Joagulation in 7 days ‘ O a. Ammonia test, positive (nitrate penuone) 4. ramperaturc elations (a) Grows at room temnerature (b) droys best at 30° J. Ciganism no. 90 I. morph0103y II. W V l. Tesctative cell 3 rm, cocci, small I Ch 2. Motility, slight if any H. 1 b. staining, Gram negative iltural Jharac aristics l. Agar s rolte (a) Jrowth, moderate (b) Jorm, filiform (c) elevatiin (d) Luster, rlistcnir; (e) Jhromogenes s cream to white. a. Gelatin (a) Growth, uniform (b) Line of puncture, filiform (c) Liquefaction, none Potato C21 0 (a) Growth, moderate (b) monography, smooth (0) Jolor, cream white (d) Luster, glistening (e) Potato, not changed 4. near colonies (a) hoderate growth (b) Ledium size 5. Gelatin colonies (a) small colonies (b) E0 liquefaction in 1 month III. nJuiOlO ical oiaiactcsistics 1. 3e entation tu‘o oes brom cresol purple plus ( has none (3) Jentrose ( ( acaction, acid A ( Gas, none (b) Lactos e( ( ieacticn, acid Gas, none (0) baccharose ( Reaction, acid 2. Ammonia test, positive (nitrate pegtone) 5, Temperature relations (a) Grows at room texperature 91 v. _‘ Q I ' v L“Sa-11811 1J0. I . Llorphol 0.6;}; l. Vegetative cells (a) form, small s7ort vols (b) size, 3 x 1.5 Ticrons EndoSpores, probably centrél . Lotil stain A‘gar (a) (b) (c) (d) (e) (f) '11r (a) (b) ity, non-motile , Gram negative stroke r . . 1,- J. Jrowuh, moderate Earn of grow-h filiform Elevation, flat Luster, glistening .L. 10pography, smooth or. romogene sis , uhi te in stab line of luncture, filiform Liouefaction, crateriform to stratiform Potato (8) Nutri A ~ ,‘ 558i 3: r'owth smooth Dolor, cream Luster, glistening Potato , not changed ent broth, slight growth colonies small white Gelatin colonies (b) Jomplete liouefaction in 1 month 1. jcrmentaticn tunes, bros cresol purple fills .‘ , ‘1 ( Gas, none (d) }l;cerin ( ( -icacti on, lrditféil s. litnus milk (a) acid (b) Joa3u1atiin r ' q U . 131.“..‘27'101’11 J“ ’3 53‘4” ““ ‘ U‘JU Ll J C U} H. tive (nitrate peptcne) (.33 J. 4. '“T78”"‘LC3 relaoi’nis (a) Grows at room tez‘njerat are (b) Grows best at 50° C. 76 p—J ‘ O O O 1‘: 31111 em I . 1.101‘pl1. 31 0,537 1. Vegetative cells, cocci, me 117 N O 5—: O r)- p. 3.1 p. ('1‘ t1; . :25 \J :3 l :3 0 rt (4. 3. Sta ning dram stain pos tive II. Jultura Jhara cte ristics l. Agar stroke (a) Growth, moderate (b) Eorm, filiform (c) elevaLion, flat (d) 10710.311:'h;,, smooth ( e) Shrcm03enes1s, cream 8. Gelatin stab (a) Growth, unijorm (b) Form, filiiorr (c) Liouefaction, saccate ‘ \1 . Eotato (a) Growth, moderate (b) color, light yellow (0) ”fl-38.13] . 4. Nutrient broth (3) moderate (a) Growth, moderate (b) Eorm, circular (3) Surface, smooth (d) JlOVation, flat (e) edge, entire A Fig V Luster, dull 6. Gelat‘n colonies (a) Small (b) Eartial liquefaction in 7 days (0) complete liquefaction in 1 month III. Physiological Jlaracteristics 1. Fermentation tubes, bron cresol purple plus ( Gas, none (3) Dextrose ( ( Reaction, abid Gas, none ( (b) Lactose ( ( Aeaction, acid 4 - ‘11 - (c) Saccharose “A“ I r4 (9 L1 L4 ct- H 0 £3 . {:1 0 (J. p. (d) Glycerin “A“ neutral 1k *Jo a. Litmus m (a) AOid in 4 days (b) coagulation in 7 days 5. Ammonia test, positive (nitr1te peptone) 4. r1"9.t;13*,ea.7."ature r elati ens ‘ ' ~v‘ ‘\ " 7MV\~" “ “ ’ (2; Grows at room tel oratire (b) Grows best at 80’ J. 1. horpholOgy 1. Vegetative cells (a) Form, cocci (b) Size, 1 x 1.5 microns 2. Lotil ty, non—motile II. Jultural Gharadteristios 1. Agar stroke (a) Growth, moderate (b) Form, filiform (c) jlevation, flat (d) Luster, glistening (e) Ghr mogene sis , white 2. Gelatin (a) Growth best at tOp (b) rorm, filiform (c) Linuefaction, saccate in 1 month 5. Potato (a) Growth, moderate (b) IOpOgrsphy, smooth (0) Jolor, cream (a) Potato, dark grey (3) null to glistening 4..Agar colonies (a) érowth, moderate (b) Eorm, circular (c) Surfsc smooth 9 (a) Growth, slow (b) Small (0) Partial liquefaction in 1 month III. Physi0103ical dharnctorist cs 1. fermentation tubes, brom cresol purple plus ( (a) gen rOS' E ( (b) Lactose ( ( fieaction, acid ( Gus, none (c) Saccharose 1 ( devotion, acia ( Gas, none (d) Glycerin ( ( fiesction, neutral 1 5. gemperntare relations ' - ‘V r WV * “' -x ‘ (a) lrons at roon temgerntnre J (b) Grows best at So ° 3. .. ()4. - III. EhjSiOlOJI331 Jhar cteristics l. fermentation tubes, brom cresol purple plus ( Gas none (a) nextrose ( ( Jeaction, acid I" _ ' UBB, none (b) Lactose AAA neaetion, aeid ( Gas, none (0) baseharose ( ( deaction, acifi ( Gas , none (d) Glycerin ( ( Jeaction, acid {\3 o Litmus milk, reduction in 7 days 3. st test, positive in 4 days 4. Ammonia test, positive in 4 dzgs 5. fienperature relations (a) Grows at room temperature (b) Grows best at 30° 3. I. morpholOgJ l. Vegetative cells Dorm, cocci, small 2. Motility, none 3. staining, cram negative 11. Julturel Jharaeteristics l. Agar stroke (a) irowth, moderate (b) form of growth, filiform (e) Blevation, flat (d) Luster, glistening - a» _ (e) TOpOSrapry, smooth (f) Jolor, white 2.3eletin stab (a) Growth, best at ton (b) Born of growth, (c) Liouefaction, none 3. rotato, slight if any growth 4. Agar colonies (a) Jronth, moderate (b) form, circular (0) surface, Enaooth (d) Elevation, flat (3) BOpOgraphy, smooth (f) Cptical characteristics, Opanue (g) JhromOgenesis, white 5. Gelatin stab (a) drowth, best at ton (b) Line of nuncture, filiform (c) Liouefaetion, none 0. Potato, slight if any growth . Agar colonies (a) Growth, moderate (b) Form, circular (c) surface, smooth (d) Jdge, entire 8. Gelatin colonies (a) Growth, moderate (b) Small (c) Partial liouefaction in V lays, complete in 1 month 1. \V b.) iolOgical Jhareeteris Jermentaticn tubes ( (d) Glycerin ( ( '- c)o '- eas, none Reaction, acid (0 3a , none deflation, 801d ( Gas none ’ c) saccharose ( ( Reaction, acid Gas, none Litmus milk (a) Acid in 7 days (b) coagulation in 7 days Ammon a tee , positive (nitrate peptone) temperature relations (a) Grows at room temperature (b) erons best at "%° J. Crganism RC. 90 I. Morphology II. 1. Yeretative cell form, cocci, small 2. Motility, slight if any 5. staining, Gram negative Jultural Jharactoristics l. (e) chromosenesis Agar stroke (a) Jrowth, moderate (b) dorm, filiform (c) dlevation, flat (d) Luster, glistening , brom cresol purple plus , cream to white. 2. Gelatin (a) Growth, uniform (b) Line of puncture, filiform c) Liquefaction, none 5. Potato (3) Growth, moderate (b) 20pography, smooth (0) color, cream white (d) Luster, glistening (e) Potato, not changed 4. Agar colonies (a) hoderate growth Gelatin colonies C31 0 (a) Small colonies (b) Lo liquefaction in 1 month III. ihgsiolegical Jh rasteristics l. Fermentation tubes, brom cresol purple plus ( Gas none (a) nextrose ( ( deaction, acid ( Gas, none (b) Lactose ( ( leaction, acid Gas, none ( deaction, acid (0) saccharose 2. Ammonia test, positive (nitrate peptone) J.o l. fejetativ; cells, large cocci 3. Lotility slight if any 0. staining reaction, Jram Stlii negatiVe J_ . olcS ultural Jhnracteris l. ‘ear stroke (a) Growth, moderate (b) form, filiform (c) Luster, jlistening (i) Iooography, smOOth (e) Jbromogcnesis, cream (f) Edge, echinulate 2. Gelatin bBSL at tor c) linuei‘action, none Stratifor: in l montl 3. Potato .' ‘- y. L‘ .' 1“ _ ~ - 'VQ . (J) awa'ull, TQ)U.GJZ;td’ null]. (b) Lrster, cull e. Gelatin colon cs -._ m,:_. ~ .2, I... ~_, .g‘ ,~. 9' (b) raluiel liouelaoulcn 1L 7 QQQB J 111. Physiological JLEEmGtOIlSLiUS l. Jermentation babes, broa cresol ( (a) ueLtEOS‘ : ( &as, none (b) Lactose ( ( .Rezt:titn1, .cicl " s ‘ urns , 11 371.0 I (c) saccharosc AAA )XLDple 1filus jeaction, acid ( Sas, none (d) }l;cerin ( ( -flrzctixxx, irritfu ;. litmus milk (3) acid (b) Seagulation ’1 o. Ammonia test, yo-itive (nitrate ( '1 -_ . - _ w, J. 4. lenperatiie relations 4. (a) Grows at room tenterauure (b) Grows best at 30° C. Organism No. 76 I. Morpn3105y l. Vegetative cells, cocci, me ium 2. Lotility, no ~motile 5. Staining Gram stain pos tive II. Julturel Jharacteristics l. Agar stroke (a) Growth, moierate (b) fora, filifiorm (c) elevation, flat (d) EOQOgrapby, smooth ( e) Jhrcmogenesis, cream c) «r _ '5' 0- ‘1. no J31‘d ell]. {3be (a) lrowth, uniform (b) Jorm, filiform (e) Liouefaction, saocnto J. Eotgto (a) eli¢htly spreading (b) Creamy (3) Normal potato .l ‘1, . ' ..,,‘ - -.‘ J.‘/ ‘1. i1 thi‘l GILL! bio 4;] ct- ‘ i) (a) Glofiding, €115“ (b) Seliment, viscii 5. Age: colonies (a) aront , :arid (c) Alevatian, flat (d) Jigs, undulate III. Physiological Jharactoristics l. Fermentation (a) nortrose ( " “ gas, none xeacticn, acid ( (d) Glycerin ( 3. Litmus milk (a) Joaguletion, smooth in 9 days (b) Reduction in 3 days 5. Indol, positive 4. Ammonia test, positive 5. Diestatic action, positive 6. Temperature relations (a) Grrows at room temperature (b) Grows beat at 50° C. (c) Grows at 45° 3. . .0-,,, ., Organisn no. 31 ibis o- anism is similar to L0. 17 except m 0] be 0 1...! H J I I H O morpholOgy l. motility, non-motile II. Phasiological Jharacteristics ' _. LAL r‘A l. Fermenuaui.n "\ .0 b (J 1" c?- H 0 C1 CD ARI—K (b) Lactose “a“ g Gas, none ( Reaction, acid IlOlla Gas, A p. v Q) (.1 K; C.) (D *S P. pd H “ma-I». negation, neutra ibis organism died bejore I had completed my studj; therefore I could not check my results. I. norpholOgy - as follows: (a) Short tlick rods (b) Size 1 x 1.5 - 2 x 1 micron ( 0) True motility (d) Gram rositive 5. )ellulose Decornosition, —. All of the organisms were tested for cellulose de- 10moosition in the following way; at ips of filter oaner me; a cut to fit into test tubes and these tubes were filled with Dunham's ablation and sterilized. rue tubes were inoculated with each culture; one was left and called aerobic (ccmditions similar to those in other tube had sterile oaiau fin oil pllt on ton :1 l L.’ (u etting ); H of the liquid (anaerobic). robes with controls were placed at 50° 3. for four: months. from all Gulo'MIO both aerobic and anaerobic (when oaper was tested with a platinum needle), the res lts were negative. 4. )ulturel studies on Slnthetic hedia. some of the typical organisms were t ici on vari 111s kinds of s t11o tic media. In th (J n first 1.,1laoe the kinds of media listed in the Iable of contents were used in licuid form but it was inn ssible to tell anything about growth, due to sediment of chemicals and pectin,even when smears were made. “hose L10 uia were made as follows: (a) Usc11i11m1y’s heparagin medium. lCOO c.c. 'schinskj's Asparagin medium as given in Jiltner's Lanual was used as the basic substance, and 1.5 percent fermented a 3a1 was added. lhen this solutign was divided into four parts ‘s1a1a5in medium plus agar pH 3. Uschinsky's asearaain medium plus agar ph 1 percent pectin. uy's As13aragin medium plus {:1 011' :13 *1 (>1 0 C.‘ U] (3 51 H. p. ,-J (‘54 I I and made into the following: (natural DE). A d. Uschinsky‘s Asparauin meiium plus agar (natural pH plus 1 percent pectin. This gives four different combinatiuns irom medium. Jhe pectin used was DOG will '. (b) Jonn‘s solution as -he great up as above, but due to haOH was added, the medium was adjusted to pH 6.8. (c) winOgradsky's medium for symbiotic given 'n liltner's flannel a derman product, probably "free was made ar,1.ount of 17‘e1,i1,itate 1' hen nitrogen fixation combinations as given in Jiltner's manual was made up into four as described previously and half of it adjusted to ph 7.2. (d) donn's ASparaginate agar. Agar 30 gms. Sodium as} are ginate 1 gm. Lcnobasic ammonium ph083hate (LHQH P01) 1.5 gm. 383 2 0.1 gm. Mgfioi 0.3 gm. 19313 t 1 e ch 1000 0.0. it: . Half of this was adjusted to pH 6.8 and the other half to pH 7.3+; to tLis last was added 5 gas. of nectin. The agar was washed in distilled water arfi.allowed st stand over night. (e) Jonn’s agar for actinomycetes Agar 30 gas. ml; 38:. in 10' 33:3. sodium asparaginate 1.0 gm.. Lonobasic as monidm (hydrog n) phOSphate 1.5 gm. hgdoé ‘.2 gm. )alciam Jhloride 0.1 gm. Lerrio Jhloride trace Half of this was adjusted to pH 6.8, the other half to pH 7.3; to this last was added 5 gas. of rectin. (f) Jarret-juice agar. Jarrots were cleaned and prepared, out into small pieces and cooked in water as foll‘ws: 250 gms. of raw carrot ylus 500 0.0. of distilled water. lhis was added to a half-liter of 3 perce‘zlt fe;~cented agar plus 5 gms. of peptone and 2 1/2 gms of sal t, and adjzsted to 3,11 7.0 desalts. It was fedud.that in some cased when pectin was added the -3diam would not M011 if #30 whis was pi~obablv due to the 2 - . ”a m 41,- !J‘ . '1, "1 4‘ ruJ‘---‘.1 . ‘ '3‘,‘ fl '1" ‘ .5 acid iormex M581 use ectin haw steiiiized. ia3:eiere, 1 D81 the £0113 :in0 solid me die to ivork with: Synthetic fledia. Lutrient agar. III - IV - VII - VIII - \ Varrot ‘ s ‘ ~fi-fi '? U luJ- Q ‘ .‘s I ‘ u 014.1". 0 | ~ '1 Jonn'b agar. agar for astinovgaatas,pfi 7.2,p1us pectin. agar for actinomywetes, UK 6.8. l .‘N- . J— r r) 8Jp8r831n8u8 agar, ‘u 7.“, p113 solution, ph 6.8, Flap pestin. bolution,pfi 6.8. golat 32, Tu 5.8. ‘f a ‘n . - 1..."? ' ‘ .I ‘ T ‘-‘ A -:Jb3glflbuyro aurarabln agar, pd ..H. " -.‘ ‘-\‘ ‘1 v 1 O ‘ ‘ fl , - ”liobradpx '5 medlun, pm v.4. ' . ‘a ~ -. 1 v ' V var ~ :7 n 2.1 " nllllr);3‘:3 (18-: E; 1nd (111111, 13;; '7. Cl. / -,. . A - . r" 4' v" ~q1v :5 I 1 4‘ .- u3t Lav 215 tlde uO we“. Mb all 0 I u- 3 * v ‘ VA . ' 'A '~¢ .' +- OffiLulSlb I 43131 tyhical sod-shaped 0?gtnis.s to pick out six of W‘ I - 1 (’1.an .L ,_,—...-r f. v I I I I I I I T I I I I I I I I I I n0 0. p . ~ . _ _ _ - . a d X e E +U I I I I I I I I I I I I I I I a S 2 a]. 1 o u 0 UL . . — _ . u _ . . . . . C r K 0 .Tu Tu n n I I I I I I I I I I I I I I I .l O 1 n C . . U .04 p . . . . . — . . ~ . . . . n . x E I I I I I I I I I I I I I I I I I I no . h . + + + + + + T +_ + t 2 .0 + + + + W X + 0 ml 0 V r O O I I I I I I I I I I I I I I I . HO N 2 . h o + + + + + + + + + + .Tu t m P + .T + + + -T + h .x S X .T + 8 h 0 .1 ”PM .1 L...” r n I I I I I I I I I I I I I I I 1 V «U a l S O 00 o + + + + + + + + + + + T r P + + + . + To g 0 X + Wu. “LL 6 O T I I I I I I I I I I I I I I I I I v n n0 0 + + + + + + + + + = = 5 p + + + + + + + + + X + + .. +_ . f o E O O I I I I I I I I I I I I I I I .me 2 h o + +. + + + + + + + + + t m P T + + + + + + W S X + .T + T o 1-1 r m .II I I I w- I I I I I I I I I I G a 1 g o + .T + T + + + + + + + r .04 .T + + .T L... .T .T O X .T + + T 7. r. I I “I... I I I I I I I I I I I I I I n0 . .T + + .T + +_ T + .T T a ”D p + + + + + + + + + .1 X + «3. “I o a.“ e O O I .I I I I I I I I I I I I I I I I I I I I I I I I T I I I I I fl. I I I I I w. 2 .. . L L J. . T + + + -T T T + + -T + C .T.v m .0“ + + + + +. + + + + + + .1 W E X 4. ,3. O .1 .M e r n I I r I T I I I I I I I I I I. c\ L. h . G a l t go o + + + + T + + +_ + .T + 1 I my + +. + + + + + + Wu 0 L + S E 14 L; t - - , I I I I I I I I I n m0 a I m1 0 5 L. + + + + + T + + _. . + + no I .T + + T + + + S I -T m. f . E (u nu O I I I I I I I I I I I I I I i E 2 ..,.L h . + -T ,T + + + T + T T + u. ,3 1 m-.. + + + T + + + + + T .., W S .3” m- h .fl 3 C .1 H .L t t flv mi 21“ I I T I I I I I I I I I I I W Wu h . me Q l O O t X S .. + T_ +_ .T T + + . + -T + + r T W .1 r . ,..__ + + + -.r + + + 00 on O M O .4 + r “14.. t B at. an-“ r: I I I I I I I I I I I I I I I I m. 4.0 a . + L a t o . T + + + + T . + + + d r h h 5 E T T .T T + + + .1 0 TL +.L f + m d .1 W O o .y . TD 0 l ‘U 0 I I I I I I I I I I I I I I nu m S I..-” L... ,2 C t L . + .T + + T + + + + T -T = __ = W m _..L .T + T + T + T + T N...V Um“. L a IT LT + + P. .1 L . + + an...“ MA I I I I I I I I I I I I I I + L 1..“ C . T T + T + .T + T + + .T T 0L -T + + T + + + + + I. .v .._ u .fU 1..” .7 CU j I 1“. My I I I I I I I I I I I I I 2.. . L «C . ...- + 1 LT _ + + . + + + — LAX. + T + LT + + + + o . w + f O .. O _-... I I I I I I I I I I I I I I I 3. ...L. S o T ._. .T + + + + .T + -T +... .u _ T. + + + T + w” S L + .U .1 . 1" ML. n I I I I 8 I I I I I I I I I I N: . 1 . r...,.L,. O + .-. T LT LT LT .T *- LT .l ._,.. r + T + T T . + + o a + I I I I I-_ I I I I #- I I I I I T I I I I I I I I T I I in I I I I I f. I I fi- I I I I I f .74 C n . at m. 45 S I i a. O T L. C .Q. I E r ..W I I V V. I I I .7. X I Q . L Tu. g r 5 I I I V I I I X m _ “ flu V I! |IIJ IV .LJ‘ILJJ‘~ (‘ f‘ 3-". i o _. L, L1. ' i q .1 ,' 1‘ 13: Li C O ‘3 inad801at0 kuowlolge HS 'W‘fl'f, div-r36: I"; an ,t 3.1. 0141.11- n.) a.) l U141: .L‘ 0 - ’wr- U -119 about Subject difi i O 0} 'b nodded to the 0011 r3ani_sns stii ‘“Oaps as follows: Groa‘ I ingluded r‘ 11181118 3.) :sLlLL .10 7H: 68. 033' and 41 f:om Irist fiber 1’ 9 T» 11‘]. J. S C) to FL .- ., - --.1. -.., 3:01) GOJUEiJS rounded e11ds,'ceeburiz1; io which 1" C“; 633 311 Spores whey are all Gram JOSiti of them liquefy gelatin. and vary as attack starch. availifbla on "Grunt media do to ac tion on mi n 2‘. ‘Tu 17‘: p; -LQUU-..LD odeCh .... - ,1. .,, ‘I ”.01- 1;, .e. I.“ 4330:1313 0 “(iv 1.1LUOd \LJIJ U0 ULG \. the Cdltdral olaracterlstics f.‘ :r J? ,, 7 f .. on 11aL 1ibos. ha Lor mignt navaYKhnltgod.in O 9 7‘"! y': ' "(j ' 11.4. \fJ-L 14“ T‘. L) C \ nf‘“ N f“ ”.(‘L *(DJLLHLU 0 S Los. -, 10, 54 and O ,-. . are isolated from Jhilean fiber, Ja ‘ and 68 and 69 from Lishfigan finer nisns which ar= medium sized rods with 1v- about 5 A 1 microns, having Slightly not affect the shape of the rods. :1 $1-“... ”1 ve. All but 54 are very motile, 1-1 whey grow very well on totato media (V AL). k and sugar bro tF All of them In this wo;1c an atten,t was "ado to name than 9 o ganioms uith.the aid of Bergay's (16) and Jnester‘s (23) “annals. It was found, however, that the dasariptians wars too moa3el‘ . It was also found that ver 1i: lo vm1‘k has been done with aerobic Spore -be arin3 organitms. Jhe'% fire it was impossible de1initely to name any of tha organisms isolated. Numbers 68, 5, 41 and 69 -58.. are sixiler to B. r333thesinm (mergey) 31.3e t that they are not so large. These LeiaOueris 10 e throw this group into the mesenterieus-megetberiam reip (15). Green 11 inc 13108 Los. 83, 2, 38, 55 end 15. Organisms 33, 2 ani 15 w3re isolet3d from Chilean fiber, are 35 and 58 from Iris} fiber. whis 3roup cont mils organisms which are medium sized rods uith rounded ends, meesurin: about 2.5 X 5 microns ! and having eeentrie srores nhioh cawz e the rods to appear ,1 .1. ,. '-‘ - .. 4 ° : 1’ s ab-sneneu. 1.933] 813 Grafi no itive emu ‘J 1 cases there is 1'18 3-!- tr3e motility. Gelatin is lidlefie d; gro‘:fih on potzto is moderate 1y abufidant .0 the 0:3 n'ene vary as to ac tion on milk and sugar 25, 53 and 13 belon; because the shape of the Sporulatir3 rod is not deiinite. in these cases the or3znisxs seem to be 3130— v - .‘3 . '; \ I'Vv‘ ”'L“ , l)" -~-- .- ‘v-Is - '2'\f-' - " q >v 1:103:0131'3 (~1). ume‘éio :3, £3.) 22110. ()8 whfi,’ .LL Jul 3230.8 1131: twollen to sli3htly club-sLaped :OdS, which M0111 place tbeS3 organisms either in Group II,or the megatherium-mes enterieas group (Jer3ey) (15). Numb r 15 varies from spindleushaped to club—shaped. If they are considere delub-shaoed, theJ fal into the group which, 333 ordin3 to $ergeJ (16) included §.3£seadotetanious, B. terminelis and B. mac ens. ihev wauld fall into G1 on _ _ ____________ d VII (Leubaeh Jiee and 30rd) (10) tee round termirsl—eeored 9 ’ , ‘ group or into 3-:oup VIII (Laabeeh, dice ena Lord), the OJJiLdri- fioation they would fall into 313 as XIX, the tetanus type, uhieh included 3. sublenetus and 3. ggtrifieus. KO '4 V Groip III includes hos. 6 d end 35. Organisms 6 and 9 4 were isolated from shileen fiber, and 5: from Irish fiber rhe organisms of this group are ”edium sized rods tith rounded ends, 13 asuring about 3.5 x 1.5 1cron Levin: centrally 10- }J0 cetcd Shores. Some of the snore—hearinr rods are s, .L .. -J .5 ndle-sheped. whey are all Gram positive and have true motility; they linuefJ A 33 Metin there is little ifs Mlj greets 01 notato‘ mill: is 33st torize-d by all; there is 3 Variation in tleir ssger reactions: they give a oositivc die ste tic test. Due to the pleomorphic characteristics of these organisms, as in Group II, their identity is doubtful. Due to the close res emblznc cc to choides and. subtilis theJ seem to belong to this group (16). Grout 11 includes hos. 91, 78 end 72. Orvcnism 91 was isolated from Courtrei fiber, 73 from Lichi3sn fiber and 78 from Jepenese fiber. ibis erode contains organisms which ere small rods measuring about 8 x 1.5 microns. gheJ heve s30 es which are "3‘ very small and centrally located. Hunts rs :1 and 78 are gram negative end 73 is Gram positive. All of the organisms in tlis 3r01p e3 cept "’ are non—motile: gelatin is linuefied bJ $1 aid 78 but not bJ 78; the orgen ens grow very well on potato; they w- ‘ vary as to reactions on litmus milk and sugar oroths. whese seem to belong to the group having 3. suhtilis as type species (15) 3513 1pleces 72 in the aerobic motile group and 91 end 78 in the aerobic non—motile group. Group V consists of one rgenism, No. so, which was isolatel from Irish fiber. It is a slender rod with round ends, measurin ‘ about ) x 5 microns. 1t is not a spore-producer, is 19:. ’ 4— 0313 81 ightly motile and s 1ram nccstive. lu doxerun livuefy - 60 - gelatin and grows well On potato. It reduces litmus milk and produces acid in dextr se, lactose, saccharose and glyeerin broths. rsnisn belonms to the TenilJ bee eriaceae (16), Q \l Tribe Jbromobactereae and ienus Flavobacterium, but due to lack of description it was infossible to trace this organism further. lroue 71 includes 30s. 8;, 90, 79, 76 74, 19, 2, 26 and 17. Numbers 12, 19, 17 and 26 were isolated irom shilean 76, 7C and 83 from Japanese fiber ‘h a ‘ ‘ ‘1" . 1 O' U ‘ | 4' 7‘ O " \ , ’9 4". ,’ 2" ‘ 2‘ 1"” ' V" ‘v‘ . ‘- f‘ "\ "th 1103:. Lilli; 15.3311.) Conneilhb 811 J; bile COJJJEJ #01145). I’Sflnxlbl’ub v 26 and 17 are ersm fOsitiVe while 90, 79 and 12 are drum negative. Numbers 73, 90, 36 17 and 21 do not liquefJ gelatin, while 83, 79 , 78, 7“: l‘.’ and l2 do. L‘hesa organisms vary in ' J“ - fl . J‘r. \ . -‘ ~fl~ V- ' 1 4‘ fl ' "‘ I r? . -.,Q '-' 1'- aceian on liumLs milk 331 sugar brothe. ineu all 3103 hell on .. . , .c 1-1, M . , -1! 4.1,, , - , .9. . , 4. .1. ., It 3-s 131nd net .oee or she 4:3en1sns Jested mould . ‘ . ., 4.1 . 1-1-.. . i, _., ,1 deeombose cellullse bx UCD Debqu used. crow well on synthetic media as l C) . .3 J_'! .- m H“, o H 5L1]. 0-- 0.41123 OL‘:5.-111DXS ‘A, .. 0 ‘,,fi , mlruinxlibele I. V ,‘I‘ ‘I‘ T ’ .' T7 ' '"V I D \J.L..L.L..L.‘XJ.LJ. 'z‘ , . - , ‘ .- .. . 0 _, . r, _ _ : ,W F -9 , ,7! ., J," 7,. 1. ;ne x93¢031u13t141§ ufpffiliuflfi --01nfl.;;1 s 3}. ,3; ‘ R "o ,._r.-, .5 I .>‘«. s 5'?-‘. V. ' h‘” ~41 .‘H ' , . _ ' 3' S" ' ‘ .. ‘ ”\fi" '.". ". ,f ‘9 -" .' v I '. .5 .' a . ‘15-» ‘r.‘-‘, fi‘ “‘- I {34’ o ‘- .“‘...‘-r’ L}: \- . ‘ "Fvif.t ~ .‘. I I , . H ,“ _ . . -: ‘7 IL "-i " _ . “,,- ' A ‘S‘fl.’ \ ‘ . 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