STREPTOMYCES ALBUS (ROSSI-DORIA) WAKSMAN ET HENRICI:TAXONOMIC STUDY OF STRAINS LABELED STREPTOMYCES

ALB US'

T. G. PRIDHAM AND A. J. LYONS, JR.

Fermentation Laboratory, Northern Utilization Research and Development Division, Peoria, Illinois

Received for publication August 24, 1960

In streptomycete taxonomy and nomencla-ture the most important species is Streptomycesalbus (Rossi-Doria) Waksman et Henrici. Itis the type species of the genus. No type strainof this species has yet been designated nor arethere any of Rossi-Doria's original cultures ex-tant. The descriptions of this species that appearin a number of publications suggest that one ofthe strains isolated by Waksman and Curtisor Waksman could be designated as the typestrain.

There have existed, since about 1916, twoentirely different concepts with regard to thenature of Actinomyces (Streptomyces) albus. Oneconcept centers around strains with the follow-ing characteristics: flexuous fruiting bodies,colors of aerial mycelium in tints and shades ofolive-buff (yellowish gray or tan); nonchromo-genicity (inability to form brown, deep brown,or black diffusible pigments in organic substrata);and marked abundance in nature. The otherconcept concerns strains that are characterizedby coiled or spiralled fruiting bodies with caten-ulate ovoidal spores; by aerial mycelium colorsgenerally interpreted as cretaceus (chalk-white,often with faint tinges of pink); by nonchromo-genicity (inability to form brown, deep brown,or black diffusible pigments in organic substrata);and by their relative rareness in nature.No type strain of the species was designated,

although the collective works of Waksman andCurtis (1916), Waksman (1919), Waksman andLechevalier (1953), and the material on Actino-myces albus and Streptomyces albus appearingin the several editions of Bergey's Manual ofDeterminative Bacteriology (1923 through 1957)suggest that one of Waksman's strains mightbe most representative of the type.

I Supported in part by a grant from the Sub-committee on Taxonomy of the Actinomycetes ofthe Committee on Taxonomy of the Society ofAmerican Bacteriologists.

In an attempt to settle this point, we reviewedthe available early literature, made a taxonomicstudy of some 42 strains, and initiated prelimi-nary investigation of an additional 13 recentacquisitions. All these strains were receivedwith the epithet "albus," or as varieties or formsof albus. The results of this study are presented,and problems concerned with the concept ofStreptomyces albus are discussed.

HISTORY

To determine whether any of the prior taxon-omic work on strains of Actinomycetes, desig-nated by the epithets alba or albus, points withcertainty to any particular type of streptomyeete,we reviewed the most pertinent papers of theearly literature. Included were those of Cohn(1875) on Streptothrix foersteri; Almquist (1890)on three unnamed streptothrices, subsequentlystated by Rossi-Doria to be identical with hisStreptotrix [sic] alba; Gasperini (1889/1890) onStreptothrix foersteri, also claimed by Rossi-Doria to be identical with his Streptotrix [sic]alba; Rossi-Doria (1891) on Streptotrix [sic] albaand S. foersteri; the collective papers of Krainsky(1914), Waksman and Curtis (1916), Waksman(1919), Jensen (1931), Duch6 (1934), Baldacci(1939), Krasil'nikov (1941, 1949), Waksmanand Lechevalier (1953); and the several editionsof Bergey's Manual of Determinative Bacteriology(1923 through 1957).Review of the literature published prior to 1916

strongly suggests that Rossi-Doria, who firstused the epithet "alba" for an Actinomycete,was probably working with strains of strepto-mycetes now commonly referred to as Strepto-myces griseus. The fact that he commented onthe marked abundance of Streptotrix [sic] alba,that he recognized the ability of his strains toform concentric rings of aerial mycelium, andthat he implied that S. alba isolates were charac-terized by straight to flexuous fruiting bodies,

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aJ I1

a

Fig. 1. Illustrations of Streptothrix foersteri Cohn reproduced from Gasperini, G., Annales de Micro-graphie, 2, 449-473, Plates VI and VII (1889/1890).

all suggest Streptomyces griseus. The morphologyof Rossi-Doria's isolates is deduced from hisgeneral statements and through his reference toGasperini's work on Streptothrix foersteri (Fig.1). Drechsler (1919) came to this same conclusion,i.e., that Rossi-Doria's Streptotrix [sic] alba ap-peared to be identical with Krainsky's Actino-myces griseus (Fig. 2). The fact that Rossi-Doriareported white aerial mycelium for Streptotrix[sic] alba rather than the olive-buff, yellowishgray, or tan colors which are characteristic ofcommon strains of Streptomyces griseus, can beexplained. The substrata used by Rossi-Doriaand other early investigators contained materialsthat are not too conducive to sporulation andthat probably would lead to the formation ofwhite aerial mycelia with any number of differentstreptomycete types. Also, "white" has been avery loosely interpreted color designation inActinomycete taxonomy.

In support of these statements, more thanhalf the strains of albus we have received froma variety of sources clearly fall in the S. griseusgroup. But developments in streptomycete tax-onomy have come about rapidly, and most paperson taxonomy published prior to 1916 have onlyhistorical and nomenclatural interest. Had Krain-sky given any attention to micromorphology

and carefully considered Rossi-Doria's and Gas-perini's papers, there now would be fewer ques-tions with regard to Streptomyces albus.Around 1916 there was a marked change in

concept with regard to Actinomyces albus thathas been preserved in publications to the presentdate. The work of Waksman and Curtis (1916)and Waksman's subsequent revision in 1919formed the basis for this concept. Data in thesepapers presumably were used in preparing thedescription of Streptomyces albus in Waksmanand Henrici's paper of 1943.Waksman and Henrici's proposal of the genus

Streptomyces in 1943 states:"We have selected as the type species of this

newly-named genus, Streptomyces albus (Rossi-Doria emend Krainsky) comb. nov. This specieswas formerly known as Actinomyces albs Krain-sky and first described as Streptothrix alba Rossi-Doria. This is one of the commonest and bestknown species of the group, and while it maylater be subdivided into further species, it is atpresent as definite as any others. It has beenrecently studied intensively by Duch6 (1934)and by Baldacci (1939). It is colorless with whiteaerial mycelium, forming ovoidal spores in coiledchains on lateral branches of the aerial hyphae.It is proteolytic, liquefying gelatin and peptoni-

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Fig. 2. Illustration of Drechsler's Actinomyces X [Streptothrix alba Rossi-Doria; Actinomyces grisentsKrainsky (?)] reproduced from Botanical Gazette, 67, 147-169, Plate IV (1919).

zing milk with the production of an alkalinereaction in the latter. It does not produce anysoluble pigment either on an organic or syntheticmedium, but does produce a characteristic earthyor musty odor."One can easily appreciate the confusion that

would result were specialists in the field to recog-nize the original concept of Streptotrix [sic] alba.The name Streptomyces griseus has become soentrenched in the minds of specialists in thefield, and cultures of organisms that fall in theS. griseus group now are so readily recognizable,that it would be senseless to change this concept.The most logical and practical solution to thisproblem is to assess Streptomyces albus in termsof the 1916 to 1919 concept. In doing this, how-

ever, one must keep in mind that many publica-tions on Actinomyces albus or Streptomyces albusare concerned with organisms in the Streptomycesgriseus group. No generalizations on this speciesshould be made unless the particular strainsstudied are clearly detailed as to their micro-morphology, color of sporulating aerial mycelium,and inability to form brown, deep brown, orblack diffusible pigments.

MATERIALS AND METHODS

Strains studied. The 55 strains used in thisstudy were obtained from the following sources:Centraalbureau voor Schimmelcultures, Baarn,Netherlands; American Type Culture Collec-tion, Washington, D. C.; Institute of Vegetable

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Pathology, University of Milan, Italy; Institutefor Biochemistry of Soils, Investigative Institutefor Agriculture, Braunschweig-Volkenrode, Ger-many; Institute of Microbiology, Rutgers Uni-versity; Charles Pfizer and Company, Inc.,Brooklyn, New York; Parke, Davis and Com-pany, Detroit, Michigan; Stine Laboratory,E. I. Du Pont de Nemours and Company, Ne-wark, Delaware; Boots Pure Drug Company,Ltd., Nottingham, England; University of Liege,Belgium; Institute of Applied Microbiology,University of Tokyo, Japan; Colonial Micro-biological Institute, Trinidad, British WestIndies; and Oswaldo Cruz Institute, Rio deJaneiro, Brazil. We are much indebted to thevarious specialists who supplied us with theirstrains. The numbers assigned to these strainsby the contributors are listed in the presentationof results.

Methods. The general procedures used in thisstudy have been described elsewhere (Hesseltine,Benedict, and Pridham, 1954; Pridham andGottlieb, 1948; Pridham et al., 1951, 1957, 1958;Tresner and Danga, 1958; A Manual of Methodsfor Pure Culture Study of Bacteria, 1946; "Meth-ods for use in cooperative studies on criteria fordescription of the streptomycetes," June 1958,unpublished; "Final plan for the internationalcommon experiment on characterization ofstreptomycetes," December 20, 1958, unpub-lished).

Stock cultures and inocula. On receipt, soilcultures were prepared with each strain follow-ing the method described by Pridham et al.(1957). The air-dried soil cultures were used inall subsequent work. For inoculating the variousmedia, two wet loopfuls (loop, 3 mm i.d.) contain-ing about 90 mg of soil culture were transferred to10 ml of tryptone-yeast extract broth (Pridhamand Gottlieb, 1948) in test tubes (25 by 150 mm).The tubes were placed on a Gump rotary shakeroperating at 200 rev/min and incubated for48 hr at 28 to 30 C. About 0.2 ml of the resultantculture was added to each agar slant or dish.For dish cultures the inoculum was streaked ina cross-hatch design on the agar.

Determination of spore morphology and natureof spore surface. Spore morphology was deter-mined by examination of Drechsler impressionslides of strains that had been cultivated oninorganic salts-starch agar for 14 days at 28to 30 C. The material on the slides was fixedby immersion in 95% ethanol for 10 sec, stained

with 0.02% aqueous methylene blue, and ex-amined under oil immersion at 1,800 X. Anelectron microscope was not available, but dataobtained in other laboratories on some strainsselected as representative of S. albus are includedin the results.

Determination of micromorphology. To deter-mine micromorphology, each strain was culti-vated on petri dishes containing Czapek's solu-tion agar, or inorganic salts-starch agar (Pridhamet al., 1957). Also utilized was the glycerol-as-paragine agar recommended for the internationalcommon experiment on characterization ofstreptomycetes. Our interpretation of the direc-tions for preparation of this medium follows.

International glycerol-asparagine agar. L-As-paragine (anhydrous basis), 1.0 g; glycerol, 10.0g; K2HPO4 (anhvdrous basis), 1.0 g; distilledwater, 1,000 ml; do not adjust pH. The pH ofthis solution is about 7.3. Add agar, 20.0 g;melt by steaming at 100 C for 15 to 20 min,dispense, and sterilize for 20 min at 121 C. Thefinal pH of the medium after sterilization withoutagar is 7.8. The final pH of the medium aftersterilization with agar and solidification is 7.4(greenish blue color with bromothymol blue).

After incubation at 28 to 30 C for 14 days,each culture was examined in situ at magnifica-tions ranging from 100 X to 800 X, and the strainswere relegated to the indicated sections describedin Pridham, Hesseltine, and Benedict (1958).

Determination of color. Observations of thecolor of sporulating aerial mycelium, reverseof cultures, and color of diffusible pigments weremade on each strain with the dish cultures usedfor studies of micromorphology after 14 days.Each dish, minus cover, was held at an approxi-mate distance of 24 in. below a 15 watt, whitefluorescent lamp. Colors were first recorded usingcommon designations such as yellow-brown,cream-color, etc., and then each was keyed outto its approximate equivalent in Ridgway (1912).The Ridgway color designations later were keyedout to the approximate equivalents in the Maerzand Paul Dictionary of Color (1950). Colors ofsporulating aerial mycelia were obtained frominorganic salts-starch agar cultures, whereas thoseof the reverses and diffusible pigments weredetermined from the glycerol-asparagine dishes.Cultures were relegated to the approximateaerial mycelium color series as outlined in Prid-ham et al. (1958).

Determination of chromogenicity. In this work,

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chromogenicity is defined as the production of abrown, deep brown, or black diffusible pigmentafter 4 days of incubation at 28 to 30 C on thepeptone agar recommended for the internationalcommon experiment on characterization of strep-tomycetes. Our interpretation of the directionsfor preparation is given below:

International peptone agar. Yeast extract(Difco), 1.0 g; tryptone (Difco), 1.0 g; sodiumchloride, 8.5 g; distilled water, 1,000 ml. Ad-just pH to 7.0 with sodium hydroxide, if neces-sary. Add agar, 17.0 g. Melt by steaming at 100C for 15 to 20 min, dispense in test tubes, sterilizefor 20 min at 121 C, and allow medium to solidifyas slants.

Although this medium was recommended inthe instructions for the international commonexperiment for determination of melanoid pig-ment production by streptomycetes, it is notconsidered the best for this purpose. Culturesforming a greenish brown to brown to blackdiffusible pigment after 4 days were recordedas positive. Those forming no diffusible pigment,or yellow or orange diffusible pigments, were re-corded as negative.Hydrogen sulfide production. The procedure to

test the capacity for forming hydrogen sulfiderecommended by Pridham, Hall, and Shekleton(1951) was modified in accordance with the workof Tresner and Danga (1958). Peptone-iron agar(Difco) was supplemented with 0.1% yeastextract (Difco) and cultures were examined after24 hr incubation at 28 to 30 C, and again at 10days.

RESULTS

Our examination of the 55 strains labeled asalbus, or as varieties or forms of albus, indicatesthat the characteristics of only 17 of these con-form to those cited in Waksman and Henrici's1943 paper. Of these, only about 10 appear tohave come from different sources, and historiesof some of these strains are still incomplete.It is entirely possible that all these may representas few as 5 original isolates. The histories of the17 strains, determined thus far, are presentedin Table 1. Thirty-four of the strains are con-sidered incorrectly identified. Four of the 55strains formed very sparse to no aerial myceliumand spores could not be demonstrated. Studiesare underway to determine whether any of theseare related to S. albus. No further considerationwill be given to the 34 incorrectly identified

strains and the 4 nonsporulating ones. Dataare presented principally on the 17 strains whosecharacteristics conform most closely to thosecited by NVaksman and Henrici.

Characteristics of liquid shaken cultures. Cul-tures of the 17 selected strains grown in 48-hrshaken tubes were essentially identical. Growthwas abundant, flaky, very fine, opalescent-white,and in every instance so heavy that the culturesappeared viscous. No diffusible pigments werenoted.

Spore morphology and surface. Examination ofthe literature, and our results, confirms thatStreptomyces albus forms elongated, ovoid sporeswith smooth surfaces (Fig. 3). These spores(unstained preparations) measure approximately1.25 by 1.75 ji. This information is presentedin Table 2.

Micromorphology. From observations of thepetri dish cultures it was possible to place thestrains studied into four of the morphologicalsections outlined in Pridham et al. (1958):Section Rectus-Flexibilis ............ 31 strains.Section Spira................ 19 strains.Section Biverticillus ................ 1 strain.Section Unknown................ 4 strains.The typical morphology of the fruiting bodies

of the strains selected as representative of Strep-tomyces albus is illustrated in Fig. 4. The organismforms small, compact, coiled chains of spores.The fruiting bodies occur as relatively shortbranches on rather long sterile hyphae. Theyoccur singly, oppositely branched, and sometimesin verticils which are difficult to detect. Theverticillate aspect of these strains is quite dif-ferent from that of forms that occur in the genusStreptoverticillium Baldacci.

Color of sporulating aerial mycelium. Colorsnoted after 14 days of growth on inorganic salts-starch agar of the 17 strains of S. albus are pre-sented in Table 3. The general color of the aerialmycelium can best be described as pale pinkishcretaceus. The color of the aerial mycelium ofthese cultures approaches pure white more nearlythan do any other streptomycetes we have seen.

Color of reverse of cultures. Colors of the reverseside of 14-day glycerol-asparagine agar culturesof the 17 strains are presented in Table 4. Al-though the colors of the aerial mycelium are quitesimilar, some differences are noted from strainto strain with respect to colors of the reverse.These ranged from a pale cream color to brown.

Chromogenicity. Only 1 of the 55 strains ex-

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TABLE 1Histories of strains of Streptomyces albus thatconform most closely with the description of

Waksman and Henrici (1943)

Designation HistorythsePape Received at NRRL from

American Type Culture Collection(ATCC), as ATCC 618; receivedby ATCC from A. J. Kluyver,Delft, Netherlands in 1926.

R. Gordon, Rutgers University,as ATCC 618.

T. Yamaguchi, Institute of AppliedMicrobiology, University of To-kyo, Japan, as ATCC 618.

H. J. Kutzner, Institute of Micro-biology. Rutgers University(IMRU), as FAL H78; receivedby Kutzner from S. A. Waksmanas ATCC 618.

S. A. Waksman, IMRU, as No.3004; received by Waksman fromthe Pribram Collection, originallythe Kral Collection, in Viennacarrying the label that it was

named or isolated by Berestnew.ATCC in 1956, as ATCC 3004; re-

ceived by ATCC from S. A.Waksman in 1944.

ATCC in 1959, as ATCC 3004; re-

ceived by ATCC from S. A.Waksman in 1944.

Centraalbureau voor Schimmelcul-tures (CBS), Baarn, Netherlands,as C.B.S. strain; received by CBSfrom Schmid-Kunz, who isolatedthe strain from dental caries.

E. Baldacci, Istituto PatologiaVegetale, Universita di Milano,Italy (IVP), as stiain 82x; iso-lated in 1956-1957 from diseasedbees or near beehives.

E. Baldacci, IVP, as No. 1; isolatedfrom diseased bees or near beehives and assigned strain numberF/3 and later 82x by Baldacci.

CBS, as Pollacci C13R5 strain; re-

ceived by CBS from P. Redaelli,Institute of Pathological Anat-omy, University of Pavia, Italy,as Actinodiscomyces bovis Ronchistrain; presumably first isolatedby Cortese from a case of humanactinomycosis and designated as

Act. albus n. varietas for study ofits porphyrine content.

TABLE 1-Concluded

Designation History

this Paper Received at NRRL from

C13R5-2 E. Baldacci, IVP, as strain C13R6strain 849.

F/2 E. Baldacci, IVP, as strain 52x;isolated in 1956-1957 from diseasedbees or near beehives.

A/7 E. Baldacci, IVP, as strain 79x;isolated in 1956-1957 from dis-eased bees or near beehives.

298x E. Baldacci, IVP, as strain 298x;isolated from diseased bees ornear beehives.

299x E. Baldacci, IVP, as strain 299x;isolated from diseased bees ornear beehives.

FAL H123 H. J. Kutzner, IMRU, as strainFAL H123; received by Kutzneras Actinomyces albus from Agricul-tural Chemical Institute, Tech.Hochschule, Munich.

Fig. 3. Morphology and nature of spore surfacesof Streptomyces albus F/3-2 (Baldacci No. 1 strain).Magnification, 17340X. Electron microscopy by N.Coy and W. Trejo, Squibb Institute for MedicalResearch, New Brunswick, New Jersey.

amined formed a brown to black diffusible pig-ment when cultured on the international peptoneagar slants. It was not one of those selected as

representative of Streptomyces albus.Production of hydrogen sulfide. Only 1 of the

618-1

618-2

618-3

618-4

3004-1

3004-2

3004-3

C.B.S.

F/3-1

F/3-2

C,aR5-1

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TABLE 2Morphology and nature of spore surfaces of strains

of Streptomuces albus

DesignationNaueoSprUsed in Spore Morphologya SurfacethisPaperSufc

618-1 Elongated-ovoid Smooth-walledb618-2 Elongated-ovoid618-3618-4 Elongated-ovoid Smooth-wallede3004-1 Elongated-ovoid3004-2 Elongated-ovoid3004-3C.B.S. Elongated-ovoidF/3-1 Elongated-ovoidF/3-2 Elongated-ovoid Smooth-walleddCU3RO-1 Elongated-ovoidC53R5-2F/2 Elongated-ovoidA/7 Elongated ovoid298x Elongated-ovoid299x Elongated-ovoidFAL H123 Elongated-ovoid Smooth-walledc

a ., -1 +A - +;- 1,4 - - A r- - _ - FF-44_- uur ciata obtainea trom examination otDrechsler impression slides. Examination of un-stained preparations indicates that the sporesmeasure approximately 1.25 by 1.75 u.

b We are indebted to Dr. K. L. Jones, Depart-ment of Botany, University of Michigan, AnnArbor, for this datum.

Data taken from Kutzner (1956).d We are indebted to Dr. N. Coy and Mr. W.

Trejo, Squibb Institute for Medical Research, NewBrunswick, New Jersey, for this datum.

55 strains formed hydrogen sulfide with peptone-iron agar supplemented with 0.1 % yeast extract.The test results appear to correlate positively-with those of the test for chromogenicity, despitethe fact that at least two different metabolicproducts are being measured, i.e., melanin pig--ment and hydrogen sulfide. It is not definitelyknown whether this correlation would hold trueif a large number of chromogenic strains weretested for ability to forni hydrogen sulfide. Theresults of our general characterization programof many types of streptomycetes suggest thatthere may be a few exceptions.

Ability to utilize sucrose in Czapek's solutionagar. When the 55 strains labeled albus werecultured on Czapek's solution agar for 14 daysat 28 to 30 C, some strains grew well, and otherspoorly. The 17 selected strains of S. albus grewpoorly.

DISCUSSION

The results of this study have given us aclearer picture of Streptomyces albus, providedone accepts the 1943 description of the speciesby Waksman and Henrici. With this descriptionas a basis, it was relatively easy to select strainsof the species. Studies of the physiological charac-teristics of the 17 strains, selected as most repre-sentative, should allow the selection and designa-tion of a suitable neotype strain.

Since about 1916, two entirely different con-cepts of Streptomyces albus have existed, one of

Fig. 4. Micromorphology of Streptomyces albus F/3-2 (Baldacci No. 1 strain). Magnification, 80OX;14-day cultures on inorganic salts-starch agar at 28 to 30 C.

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TABLE 3Colors of sporulating aerial mycelium of strains of Streptomyces albus grown for 14 days at 28 to 30 C on

inorganic salts-starch agar

Designation Used ComnClrNm iyDsgatin Maerz and Paulin this Paper Common Color Name Ridgway Designation Designationa

618-1 Cretaceus (chalk-white) -618-2 Pale mealy-gray to off-white618-3 Cretaceus to off-white to very pale

olive-buff618-4 Pale buff Pale Pinkish Buff XXIXc Ivory 10B23004-1 Off-white3004-2 Off-white3004-3 CretaceusCH3R5-1 Pale pinkish buff Pale Pinkish Buff XXIX Ivory 10B2C13R5-2 Off-whiteC.B.S. Pale pinkish buff Pale Pinkish Buff XXIX Ivory 10B2F/3-1 Pale buff whiteF/3-2 Cretaceus to pale pinkish buff Pale Pinkish Buff XXIX Ivory 10B2F/2 Pinkish cretaceusA/7 Pinkish cretaceus298x Pale pinkish buff299x Pinkish cretaceusFAL H123 Pale pinkish white Pale Pinkish Buff XXIX Ivory 10B2

a The Inter-Society Color Council-National Bureau of Standards (ISCC-NBS) color names (Kellyand Judd, 1955) for the Ridgway and Maerz and Paul designations are, "73. pale orange yellow," and"89. pale yellow," respectively.bDash indicates that no color tabs, other than "white" were equivalent to the color observed.c The Ridgway designation represents the closest approximation that could be obtained. All colors

might more precisely be designated as "between the white tab and the Pale Pinkish Buff tab."

which centers around cultures now consideredas members of the S. griseus group. The otherincludes a group that form coiled fruiting bodieswith catenulate, elongated, ovoidal, smooth-walled spores and cretaceus (chalk-white, oftentinged with pink) sporulating aerial mycelium;are nonchromogenic; cannot form hydrogensulfide; and grow poorly on Czapek's solutionagar. Preliminary evidence suggests that thereare some physiological differences among the17 strains of this group thit we6have acquired.The fact that ther'h'ahve been two concepts

with regard to S. atbus suggests a careful re-examination of Atheo'`synonymy of this speciesshould be made.1 .

In our; t'taxonomic -wQik on streptomycetes,we recently adoptedl&tpifactice of using distilledwater in preparing all"of our media. This stepwas taken to eliminate the effect of inorganicsalts of unknown composition. If this practicewere adopted by other laboratories, it shouldtend to reduce the possibilities for interlabora-tory differences in results of various tests. Some

differences in results now noted could be at-tributed to variation in composition of tap waterobtained from widely differing sources.

ACKNOWLEDGMENT.-

We are indebted to Miss Collette Foley fortechnical assistance with certain portions of thework reported here; to Dr. Charles Drechsler,Mycologist, Vegetable and Ornamental ResearchBranch, Crops Research Division, AgriculturalResearch Service, ,U. S. Department of Agricul-ture, Plant Industry Station, Beltsville, Mary-land, an&to the Botanical Gazette for permissionto reprodluce the material. shown in Fig. 2; ,toDr. N.' Coy and Mr.' William H. Trejo, TheSquibb Institute for Medical Research, NewBrunswick, New Jersey, for preparation of theelectron micrograph illustrated in Fig. 3; and toDean R. E. Buchanan, Iowa State University,Ames, Iowa, and Professor David Gottlieb, Uni-versity of Illinois, Urbana, Illinois, for reviewingthe manuscript during its preparation.

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TABLE 4Colors of reverse of strains of Streptomyces albus grown for 14 days at 26 to 30 C on international

glycerol-asparagine agar

Designation Us d Common Color Name Ridgway Designationec Maerz and Paul Designationbe

618-1 Pale orange-brown Pale Ochraceous-Buff XV (No name) 9C4618-2 Orange-brown to olive- Tawny-Olive XXIX to Hazel 13J9 to Acorn 15E7

brown Saccardo's Umber XXIX618-3 Orange-brown to pale Chamois XXX to Warm Raffia liE5 to Nankeen 10F3

olive-yellow-brown Buff XV to Buckthorn to Chipmunk 13L9Brown XV

618-4 Light brown Tawny-Olive XXIX Hazel 13J93004-1 Pale cream to pale olive Light Buff XV to Deep Ivory 10B2 to (no name)

Olive-Buff XL 12C23004-2 Dull cream to pale orange- Cream Color XVI to Pale Straw 10F2 to Polar Bear

brown Ochraceous-Buff XV 9B23004-3 Light orange-brown Cream Buff XXX (No name) 10E3C,aR5-l Pale yellow-brown Naples Yellow XVI Nankeen 10F3Cc3Rs-2 Light olive-brown Buckthorn Brown XV to Chipmunk 13L9 to Olive

Dresden Brown XV Brown 15H7C.B.S. Pale cream to dull cream Light Buff XV to Cream Ivory 10B2 to Straw 10F2

Color XVIF/3-1 Pale brown Buckthorn Brown XV Chipmunk 13L9F/3-2 Pale orange-yellow-brown Buckthorn Brown XV to Chipmunk 13L9 to Hazel

to olive-brown to pink- Tawny-Olive XXIX to 13J9 to (no name) 10D4ish-buff Light Ochraceous-Buff

XVF/2 Dark brownish-yellow Warm Buff XV Nankeen 10F3A/7 Olive-brown Warm Buff XV Nankeen 10F3298x Pale olive-brown Cinnamon-Buff XXIX Toltec 11B7299x Pale olive-brown Cinnamon-Buff XXIX to Toltec 11B7 to Hazel 13J9

Tawny-Olive XXIXFAL H123 Pale olive-yellow Pinkish Buff XXIX Vanilla 10C3

a Results presented are those obtained by two different individuals and reflect individual differencesin color interpretation.

b The Inter-Society Color Council-National Bureau of Standards (ISCC-NBS) color names (Kellyand Judd, 1955) for the Ridgway and Maerz and Paul designations are given below;28. light yellowish pink 80. grayish yellowish brown29. moderate yellowish pink 86. light yellow57. light brown 87. moderate yellow71. moderate orange yellow 89. pale yellow73. pale orange yellow 90. grayish yellow74. strong yellowish brown 94. light olive brown76. light yellowish brown 95. moderate olive brown77. moderate yellowish brownNote the broader range of colors as compared with those of table 3.c Results presented are those obtained by two different individuals, keying out together, the color

tabs in Ridgway indicated in second column. When working together, there was a considerably greaterdegree of agreement on color interpretation.

SUMMARY entirely different concepts regarding StreptomycesA review of the literature and a taxonomic albus have existed since 1916.

study of a number of strains of Streptomyce8 albus The earlier concept suggests recognition ofhave been made. The results indicate that two some member of the Streptomyces griseu.s group

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as the neotype of Streptomyces albus sensu Rossi-Doria and other investigators.The other concept centers around strepto-

mycetes that possess these characteristics: coiledfruiting bodies with catenulate, elongated,ovoidal, smooth-walled spores; a cretaceus (chalk-white often tinged with pink) sporulating aerialmycelium; nonchromogenicity; inability to formhydrogen sulfide; and poor growth on Czapek'ssolution agar.

We believe that the second concept of Strep-tomyces albus should be accepted. This proposalis made to prevent the obvious confusion thatwould ensue were the original concept for S.albus recognized. In our opinion acceptance ofthe newer concept will impart a greater degreeof stability to streptomycete taxonomy andnomenclature. We recognize that much of theearlier literature concerning S. albus refers tostreptomycetes that now would be placed in theS. griseus group, that many strains now availableand labeled with the epithet albus are strainsthat would be placed in the S. griseus group,and that the synonymy of S. albus should becarefully examined. Specialists who must referto the literature on S. albus should be aware ofthese facts.

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BALDACCI, E. 1939 Contributo alla sistematicadegli attinomiceti V-VIII. Actinomyces albus,A. chromogenus, A. odorifer, A. thermophylus,A. viridis, A. viridochromogenus, A. hominis,A. innominatus. Atti ist. botan. (GiovanniBriosi e lab. crittogam. univ. Pavia, Ser. 4),11, 191-231.

Bergey's manual of determinative bacteriology,1923 through 1957, 1st through 7th editions.The Williams & Wilkins Co., Baltimore.

COHN, F. 1875 Untersuchungen uber Bacterien.II. In Beitr&ge zur Biologie der Pflanzen,vol. 1, Dritte Heft, pp. 185-188; 202-207;Plate V. J. U. Kern's Verlag (Max Muller),Breslau, Germany.

Committee on Bacteriological Technic 1946 Amanual of methods for pure culture study ofbacteria. Biotech. Publications, Geneva,New York.

DRECHSLER, C. 1919 Morphology of the genusActinomyces. II. Botan. Gaz., 67, 147-169(Plate IV).

DUCHt, J. 1934 Les Actinomyces du groupe

albus. In Encyclopedie mycologique, vol. 6,375 pp. P. Lechevalier & Fils, Paris, France.

GASPERINI, G. 1889/1890 Recherches morpho-logiques et biologiques sur un microorganismede l'atmosphere, le Streptothrix Foersteri-Cohn. Ann. de Micrographie, 2, 449-473(plates VI and VII).

HESSELTINE, C. W., R. G. BENEDICT, AND T. G.PRIDHAM 1954 Useful criteria for speciesdifferentiation in the genus Streptomycos.Ann. N. Y. Acad. Sci., 60, 136-151.

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KRAINSKY, A. 1914 Die Aktinomyceten undihre Bedeutung in der Natur. Zentr. Bak-teriol. Parasitenk., Abt. II, 41, 649-688.

KRASIL'NIKOv, N. A. 1941 The guide to theray fungi, Actinomycetales. Institute ofMicrobiology, Acad. Sci. U. S. S. R., 148 pp.,

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Moscow-Leningrad.KUTZNER, H. J. 1956 Beitrag zur Systematik

und Okologie der Gattung StreptomycesWaksman et Henrici. Dissertation, Land-wirtschaftlichen Hochschule, 187 pp.,

Hohenheim, Germany.MAERZ, A., AND M. R. PAUL 1950 A dictionary

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PRIDHAM, T. G., H. H. HALL, AND M. C. SHEKLE-TON 1951 The identification of some

Actinomycetales with particular reference toisolates producing vitamin B12 and relatedgrowth factors. Presented at, 51st GeneralMeeting of the Society of American Bac-teriologists, Chicago, Illinois, May 27-31,1951. Bacteriol. Proc., 1951, 27-28 (abstr.).

PRIDHAM, T. G., P. ANDERSON, C. FOLEY, L. A.LINDENFELSER, C. W. HESSELTINE, AND R. G.BENEDICT 1957 A selection of media for

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WAKSMAN, S. A. 1919 Cultural studies ofspecies of Actinomyces. Soil Sci., 8, 71-215.

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