Copyright 0 1976 American SocietyforMicrobiology Printed in U.S.A.
Long-Chain
Fatty Acids of Peptococci and Peptostreptococci
CAROL L. WELLS* ANDCHARLES R. FIELD
Wisconsin StateLaboratory of HygieneandDepartment of Medical Microbiology,* University of Wisconsin, Madison, Wisconsin 53706
Received for publication23July 1976
The long-chain fatty acids extracted from the whole cells of 12 clinically
significant species of peptococci and peptostreptococci werecharacterizedby
gas-liquid chromatography. The resulting methylated fatty acid profiles (andsome
unidentified compounds) of 82 strains allowed the 12 species to be separated into four groups. Fifteen strains of Peptostreptococcus anaerobius were placed in
group Ibecause they had aunique, prominent compoundthat occurred inthe
area where a 0, to
C01
fatty acid would be expected. Group II, consisting ofPeptostreptococcus intermedius, Peptostreptococcus micros, Peptostreptococcus parvulus, Peptococcus morbillorum, and Peptococcus constellatus, produced
C14,
C16:1, C18:1
,andC18fatty acids. Peptococcus prevotii, Peptococcus variabilus,Peptococcus magnus, Peptococcus asaccharolyticus, and Peptostreptococcus
productus wereplaced in group III becausethey contained three to six
addi-tional, unidentified compounds that strikingly differentiated them fromgroup
II. Peptococcus saccharolyticus was the single species assigned to group IV
because it yielded C14, C16,
C18:1,
C18, and C20 fatty acids and a prominentunidentified peak that occurred between C14 and C16 fatty acids. This study
indicated that cellular long-chain fatty acids may be an important tool in
clarifying thetaxonomy of the peptococci and peptostreptococci.
Peptococci and peptostreptococci arefoundas
normal flora of the skin, upper respiratory
tract, oral cavity, large intestine, and female
genitalia (3); they are frequently associated withinfectiousdiseases(13, 18),and their
path-ogenicityformanis now widely accepted.
Accurate laboratory speciation of the medi-cally important anaerobic gram-positivecocciis difficult. Different species of peptococci and
peptostreptococci have similar biochemical
re-actions, andgaschromatography of short-chain acidmetabolites (C1to C8)often reveals identi-calfermentation products. Thespeciationof the peptococciandpeptostreptococci isfurther
com-plicated by theexistence ofmany strains that
donotfitthe already describedspeciesof
anaer-obiccocci (3). In addition, four of the foremost
authorities in clinical anaerobic bacteriology
differ markedly in theirguidelines for
specia-tion ofpeptococci and peptostreptococci (5, 7,
14-16).
This study presents an analysisof the
long-chain fatty acids (LCFAs) extracted from the
whole cells of 12 clinically significant species
(82 strains) of peptococciand peptostreptococci.
The LCFAs (C8 to C20) of the anaerobic cocci
havenotyet beencharacterized,and such
anal-yses have been useful in
clarifying
thetaxon-omy of other organisms (10, 12).
MATERIALS AND METHODS
Organisms and media. The organisms studied wereobtained from stock cultures maintained at the Wisconsin State Laboratory of Hygiene, Madison, Wis. All cultures were clinical isolates sent to the State Laboratory for identification over a 4-year period from 1972 to 1975. The following identifica-tion tests were routinely performed: Gram stain from agarandpeptone-yeast-glucose broth; fermen-tation tests oncellobiose, esculin, fructose, glucose, lactose, maltose, mannitol, and sucrose; esculin
hy-drolysis;gelatinliquefaction; indole production;
ni-tratereduction; gas-liquid chromatography of short-chainacid metabolites. In addition, other tests, such as stimulation by Tween 80, reactions in milk and meat, salicin and starch fermentation, starch
hy-drolysis,andhemolysis, were occasionally done. All
tests were performedaccording to, and with results compatible with, the Virginia Polytechnic Institute (VPI) Anaerobe Laboratory Manual, 2nd ed. (7). Twelve species were studied: Peptostreptococcus an-aerobius, Peptostreptococcus intermedius,
Pepto-streptococcus micros,Peptostreptococcus productus,
Peptostreptococcusparvulus, Peptococcus
asaccharo-lyticus, Peptococcus constellatus, Peptococcus mag-nus,Peptococcusmorbillorum, Peptococcus prevotii, Peptococcus variabilus, and Peptococcus
saccharo-lyticus.Afteridentification, the cultureswerestored
at -70°C in chopped-meat-glucose broth. Before
analysis, each culturewasthawed and transferred
tochopped-meat-glucose broth for24to48 hat370C. 515
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The purity of each culture was checked by Gram stain. Allcultures were then transferred to 7 ml of peptone-yeast-glucose broth prepared according to the formula given in the VPI Anaerobe Laboratory Manual (7) (peptone made by GIBCO Laboratory, Madison, Wis.; yeast extract and glucose made by DifcoLaboratories, Detroit, Mich.) for 48 h at370C.
The cultures were centrifuged, the supernatant was pouredoff, and the cell pellet was stored at -70°C until the fatty acids were extracted.
LCFA extraction. Fatty acids were extracted from the wholecells according to the method of Moss
etal. (11) with some modifications. The cells were thawed, suspended in 5 ml of 5% NaOH in 50% aqueous methanol and heated at 100°C for 15 min. The saponified material was cooled and acidified with 6 NHClto apHbelow2.One milliliter of boron trifluoride methanol (BF3-CH30H, Applied Science Laboratories, State College, Pa.) was added to the saponified mixture and it was heated at 100°C for 5 min. The methylated solution was then added to 10 ml of saturated NaCl and the fatty acid methyl esterswereextracted twice with an equal volume of 1:1 ether-hexane. The combined ether-hexane ex-tracts were evaporated at room temperature to about5mlunder agentlestreamofdrynitrogen and anhydrous sodium sulfate was then added to remove any water. Aprecipitatefrequently formed during the evaporationprocedure and was removed by cen-trifugation at 1,800 rpm for5 min. Evaporationof the extract withdry nitrogen was continued until a final volume of 0.1 to 0.2 ml was achieved. The extractwasstoredat-70°Cbefore fatty acid analy-sis.
Gas chromatography. A Hewlett-Packard gas
chromatograph (model 5710A) with automatic
sam-pler(model 7671A) and reporting integrator(model
3380A) was used throughout this study. The gas
chromatograph wasequipped with aflame
ioniza-tiondetector. The flamewasmaintainedbya
mix-tureofhydrogenand air with flowratesof 30ml/min
and 250 ml/min, respectively. The reporting inte-grator was set for aslope sensitivity of1 mV, an
attenuationof16 or32, andachartspeedof0.5cm/ min. The integrator printed the retention time of eachpeak, calculated theareaundereachpeak, or
identified each peak relative to anexternal
stan-dard. The external standard contained caprylate
(C8), caprate (C10), laurate (C12), myristate (C14),
palmitoleate (C16:), palmitate (C16), oleate
(C8:1),
stearate (C18), and arachidate (C20) methyl esters
(Applied ScienceLaboratories, State College, Pa.). Themethylestersof LCFAswereseparatedon a
coiled-glasscolumn6feet(ca. 183cm)longwitha
4-mmIDandaone-fourth-inch(ca. 0.64cm) OD. The carrier gaswasprepurifiednitrogen(Badger Weld-ingSupplies, Madison, Wis.), withaflowrateof50
ml/min.Methylatedfattyacidextractswere run on
bothapolar andanonpolarcolumn. Thenonpolar
columnwas packed with3% SE-30on100/120 Gas-Chrom Q(Hewlett-Packard, Skokie,Ill.). The
tem-perature of the nonpolarcolumn was
programmed
from110 to240°C at4°C/min, withatotalanalysistimeof45min.Foranalysisonthenonpolarcolumn,
thesamplesize was1,ul, the injection port
tempera-turewas 200°C, and thedetector temperature was
350'C.Thepolar column, used primarily for confir-mation of identified peaks, was packed with 15% EGSS-X on 80/100 Gas-Chrom P(AppliedScience,
State College, Pa.). The temperature of the polar column wasmaintained at172°C for a totalanalysis timeof50 min. Foranalyses on the polarcolumn,
thesample size was 10
gl,
theinjection porttemper-ature was200°C, and the detector temperaturewas
4000C.
All samples werefirst analyzed on the nonpolar column. Theremaining extract was diluted 1:3 in
ether-hexanetoobtainasufficient volume for anal-ysis on the polar column. Consequently, a 10-,ul sample size was needed to obtain adequate peak size fromthe polar column, whereas a 1-,ul sample size gaveadequate peak size from the nonpolar column.
RESULTS
Results of this study demonstrated that the
peptococciand peptostreptococci,onthe basisof
gas chromatographic profiles of their
methyl-atedLCFAs, could bedividedintofourgroups.
Group I (Fig. 1) contained onlyone species, Peptostreptococcus anaerobius. Fifteen strains
of thisorganismallyieldedasimilar
chromato-graphic profile which containeda
characteris-tic, prominent, unidentified peak with a
reten-tion time between C8 and C10. The average
percent fatty acid composition of this
unidenti-fied compoundwas 20% (witharangeof 2.4to
29.0%).This compound was always present and
could be usedasamarker forgroupI anaerobic
cocci since it was not produced by the other
anaerobic cocciassayed. P.anaerobius was also
unique among the other peptococci and
pepto-streptococci because each of 15 P. anaerobius
strainsanalyzed contained from 14 to 25
differ-ent peaks (with an average of 19 peaks). All
other species of peptococci and
peptostrepto-cocci studied never produced over 12
differ-ent peaks. After analysis of 15 P. anaerobius
strains, thefollowing statements could be made
about group I: (i) a prominent, unidentified
peak is located betweenC8andC01;(ii) a total of
14to 25differentpeaks areproduced; (iii)
C14,
C16,
C18:1,
andC18
areusually present; the latterfatty acidswerepresent in13 outof15strains
studied.
On thebasis of theircellular LCFAcontent,
Peptostreptococcus intermedius,
Peptostrepto-coccus micros, Peptostreptococcus parvulus,
Peptococcus morbillorum, and Peptococcus
constellatus were all placed in group II. The
average percentages ofthe LCFA composition
oftheseorganismsaresummarizedinTable 1.
All strains within group II had C16 or C18:1 as
the mostprominent LCFAs on the
chromato-gram. Group II alsoproduced smaller, but
de-tectable amounts ofC14,
C16:1,
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llJ (I) z 0 a. Uf) LLI
w
a:
a:
0
wL
a:
GROUP I
18:1
ULAJ
GROUPmf
18
14
16
18:1
GROUP EI
181
16
161
14
8
14
16:1
18 16
18:1
l
0 7 14 21 28 35 0 7 14 21 28 35
MINUTES
FIG. 1. Representativechromatograms ofeachofthefourgroupsofpeptococciandpeptostreptococcibased onlong-chain fattyacidanalysis ofwhole cells.
sentative
chromatogram
for this group isshowninFig. 1.
Table2shows theaverage percentagesof the fattyacid composition of group III organisms.
GroupmincludedPeptococcus variabilus,
Pep-tococcus magnus, Peptococcus
asaccharolyti-cus,Peptococcusprevotii, and
Peptostreptococ-cusproductus. Themostprominentfatty acids
ingroup III wereidenticaltothefatty acidsin
group 11
(C16:1,
C16,C18:1,
and C18). Again, as ingroup II, the highest peak on the
chromato-gram was consistently C16 or
Cj8:l.
GroupIll
differed fromgroupII,
however, by
thepresenceof three to six additional
compounds (Fig.
1). Thesepeaks wereallunidentified and occurredbetween C14 and
C16:1,
between C16 and C18:1,and between C18 and C20.
GroupIVcontainedoneorganism,
Peptococ-cussaccharolyticus. The LCFAs extracted from
P.saccharolyticusareshown inTable3.
Promi-nentpeaksofC16,
C08:1,
and C18weresimilartothose seen in groups II andIII;however, C16:1
was conspicuously absent from group IV. P.
saccharolyticus
contained significant amounts4,
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TABLE 1. Average percentages of the long-chain fatty acids extracted from group II cocci
Peptostreptococcus sp. Peptococcus sp. Fatty acids P.
intermedius
P.micros
P. parvulus P. morbillorum P. constellatus(15 strains) (8strains) (1strain) (5strains) (5strains)
C12 0.7 1.4
(0-3.7)a (0.6-2.6)
Unidentified 1.2 0.3
(0-6.2) (0-1.0)
C14 4.6 3.2 4.2 3.1 4.8
(2.3-7.3) (0-6.8) (1.9-4.0) (3.4-6.1)
C16:1 8.4 5.0 9.6 10.2 7.3
(4.0-28.3) (2.3-6.7) (2.9-30.1) (6.4-8.9)
C16 35.1 22.5 18.4 20.1 32.5
(13.7-44.7) (19.6-35.2) (14.2-31.5) (19.5-40.0)
C18:1
39.8 53.7 54.8 53.5 43.1(27.3-50.2) (41.5-63.2) (23.2-69.1) (34.9-58.3)
C08 11.2 15.8 13.0 12.1 10.7
(9.4-16.1) (11.6-20.7) (5.1-18.0) (8.9-12.1)
aThe numbers in parentheses are the range (in percent) for all organisms tested.
of C20 and an unidentified compound with a
retention timebetween
C14
andC16,
whichap-pearedtobe unique characteristics of this
spe-cies in group IV. Fig. 1presents a
representa-tivechromatogram ofgroup IV(P.
saccharoly-ticus).
DISCUSSION
Thetaxonomy of the peptococci and the
pep-tostreptococci is unclear. Four of the most
nota-ble publications in this area differ greatly in
thespeciationof theseorganisms. Publications
ofRogosa (14, 15),Sutteretal. (16),Dowell and
Hawkins (5), and Holdeman and Moore (7) de-scribe 11species ofpeptococci and six species of peptostreptococci, and they differon the
num-berand identity of species within eachgenus.
Sinceall of the peptococci and peptostreptococci
(exceptPeptococcus niger)areconsideredtobe
associated with infectious disease in humans, thereshould be reliable methods for identifying theminthe clinicallaboratory. Rogosa (14, 15)
listssixspeciesofpeptococciand fivespeciesof peptostreptococci. Of thelatter 11species, five species (Peptococcus niger,
Peptococcus
aero-genes, Peptococcus activus, Peptococcus
an-aerobius, and Peptostreptococcus lanceolatus)
are not
recognized
by either Sutteret al. (16),Dowell and Hawkins (5), or Holdeman and
Moore (7). Table4 presents a summaryofthe
current classification of the 12 most
generally
recognized species of peptococci and
peptostrep-tococci and includes the grouping based on
LCFAprofiles. Of the 12 specieslistedinTable
4,thereisgeneralagreement on the taxonomic
status ofonly two species, Peptostreptococcus
anaerobius and Peptostreptococcus
interme-dius. It is evident that much work needs to be
doneonthetaxonomyof thepeptococciandthe
peptostreptococci.
This study, for the first time, characterized
the LCFAs of the12mostuniformly recognized species of peptococci and peptostreptococci. On the basis of their LCFA chromatographic
pat-terns, each species of peptococcus and
pepto-streptococcus assayedcould be placedinto one
offourdifferent groups.
Asaresult ofLCFAprofiles,
Peptostreptococ-cusanaerobiuswasplacedina groupbyitself. One distinguishing feature ofP. anaerobius
wasthe large number (14 to 25)of peaksonthe chromatograms. No other species studied
pro-duced over 12 peaks. The most characteristic feature of the profile ofP. anaerobius was a
unique, prominent peak between C5 and C00.
Unfortunately, the latter peak hasnotyetbeen identified andmaybe an iso-acid.
Group II cocci containedPeptostreptococcus intermedius, Peptostreptococcus micros,
Pep-tostreptococcusparvulus, Peptococcus
morbil-lorum, and Peptococcus constellatus. The
spe-cies in group II produced characteristic
amounts ofC14,
C16:i,
C16,Cl8:1,
andCl5
fatty
acids. The latterprofileissimilartothe
profile
reported for fatty acids in some ofthe
faculta-tivestreptococci (2, 9). Some of the organisms
inthisgroup are now considered tobe members
of the genus
Streptococcus
sincethey produce
lacticacidas amajor metabolicproduct. Onthe
latter basis, P. intermedius is now
generally
recognized as belongingto the genusStrepto-coccus (4, 8, 14-16). Similarly,P.morbillorum
and P. constellatus are now
widely
consideredto be members of the
streptococci (4, 8, 16).
Peptostreptococcus micros, whichproduces
lac-tic acid as a minor product, is still
generally
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TABLE 2. Average percentagesofthelong-chain fattyacids extractedfromgroupIII cocci
Peptococcus sp.
Peptostreptococcus
Fatty acids P. variabilus P.magnus P.asaccharolyti- P. prevotii productus(1 strain) (5 strains) (8strains) cus(9 strains) (6 strains)
C12 0.4 2.9
(0-1.8)a
Unidentified 1.8 0.8
(1.0-2.5) (0-3.4)
C14 2.4 5.7 2.5 6.9
(0-3.3) (2.9-8.5) (0-9.0)
Unidentified 0.2 0.6 0.8 0.8 3.0
(0-1.0) (0-1.6) (0-4.1) (0-2.9)
Unidentified 2.3 0.7
(0-3.6) (0-2.8)
C16:1 5.2 4.7 10.5 6.2 2.3
(2.9-6.6) (3.4-6.4) (3.1-15.3) (2)2-13.0)
C16
11.6 11.1 24.7 13.0 26.8(6.8-17.3) (9.3-13.4) (17.7-31.1) (2.6-27.1)
Unidentified 17.2 13.3 5.0 9.0 15.8
(13.2-20.0) (10.3-21.1) (2.0-9.5) (2.9-16.7)
Unidentified 1.9 4.2 5.5
(0-5.6) (0-12.2) (0.5-8.8)
C18:1
43.6 36.6 31.3 49.1 13.4(38.0-48.9) (30.4-44.4) (18.7-49.2) (30.0-77.6)
C18
6.6 7.6 5.8 8.4 7.5(4.4-8.6) (6.8-8.8) (3.9-10.4) (5.6-11.4)
Unidentified 10.2 18.8 8.3 4.6 14.2
(2.9-21.4) (17.0-21.7) (3.7-12.0) (2.3-8.1)
Unidentified 1.4 4.6 0.9 0.3 7.3
(0-4.7) (2.8-6.5) (0-2.2) (0-1.1)
aThe numbersinparentheses are the range (in percent)for all organisms tested.
TABLE 3. Average percentages ofthelong-chain fatty
acids extractedfromgroup IV cocci
FattyFattyacidsacids Peptococcus saccharolyticus
~~(4
strains)
C14
Unidentified
C16
Unidentified
C18:1 C.8
Unidentified
Unidentified
C20
2.9
(1.4-5.0)a
26.5 (16.9-32.5)
12.8 (5.6-23.9)
2.5 (0-9.8) 14.3 (9.5-22.7)
18.3 (13.3-22.7)
2.5 (2.0-2.7)
5.7 (0-8.8) 13.2 (8.5-15.7)
aThe numbersinparentheses aretherange (in percent)forallorganisms tested.
consideredtobeamember of the
peptostrepto-cocci.Thefatty acidsinP.parvulus also placed
this organism in group II; however, only one
strain was analyzed and this should not be
considered a definitive characterization. The
VPI Anaerobe Laboratory Manual also de-scribes P. parvulus on the basis of a single isolate and the Manual also notes that this
organismproduceslacticacidas amajor
meta-bolic product (7). Thus, the most significant
interpretationsof the LCFA profiles ofgroupII
organismsare: (i) theseLCFAprofilesare
con-sistent with theprofilesof the facultative
strep-tococci;and(ii)allspeciesnowbeingconsidered
for classification with thestreptococci have all
beenplacedingroup II(Table 4) on the basis of
LCFAanalysis.
Group
IIIcocci, consisting ofPeptococcusvar-iabilus, Peptococcus magnus, Peptococcus
asaccharolyticus, Peptococcus prevotii, and Peptostreptococcusproductus, appeared to be a
miscellaneous assortment of organisms that produce similar LCFA profiles that consist of
C16:1, C16,
C08:1,
andC1l
fatty acidsin amountssimilartothoseseeningroupII. The
chromato-grams oftheorganisms in group m contained
from threeto sixadditional, unidentified peaks
notseen in group H organisms, and thus
war-rantedthe formation of a separate group. The
organisms in group III possessed some
associa-tions that were consistent with the published
literature on anaerobic cocci. Holdeman and
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TABLE 4. Current classification of the species of peptococci andpeptostreptococcia
Organism Rogosa (14,15)8 Sutter et al. Dowell and Holdeman and Long-chain fatty
(16) Hawkins (4, 5) Moore (6-8) acidgrouping
Ps. anaerobius R R Rc R 1
Ps. intermedius S S S S 2
Pc. morbillorum U S S S 2
Pc. constellatus R R S S 2
Ps. micros R R Q R 2
Ps. parvulus R R Q R 2
Pc. magnus Ud Rd Q Re 3
Pc. variabilus Ud Rd Q Re 3
Pc. asaccharolyticus R R Rf R 3
Pc. prevotii Ug R Rh R 3
Ps. productus R R Q R 3
Pc. saccharolyticus U U RI R 4
a Abbreviations: Pc., Peptococcus; Ps., Peptostreptococcus; R, recognized species; U, unrecognized species;
Q, questionable status; S, Streptococcus species.
b Numbers in parentheses indicate literature reference.
c Designated Peptostreptococcus Center for Disease Control (CDC) group 3.
dConsidered the equivalent of Peptococcus anaerobius.
eConsidered equivalent organisms.
' Designated Peptostreptococcus CDC group 1.
9Considered the equivalent of Peptococcus asaccharolyticus.
hDesignated Peptostreptococcus CDC group 2. iDesignated Peptococcus CDC group 2.
Moore(6)considerP.magnusandP. variabilus tobe thesame organism sincetheir
biochemi-calprofilesareidenticalifoneadds Tween 80 to
their growth medium. Rogosa (14) claims that
P.variabilus and P. magnus are the equivalent
of aPeptococcus anaerobius. As a solution to
this obvious confusion, West and Holdeman
(17) propose that Peptococcus anaerobius
should be
rejected
sinceitisa nomenconfusumdue to the existence ofPeptostreptococcus
an-aerobius. An example oftwo other organisms
thatmay notbeseparatespecies are P.prevotii
and P.
asaccharolyticus.
Rogosa (14)statesthatP.prevotii and P.asaccharolyticusare
indistin-guishable since they differ only by an indole
reaction out ofover 100 other characteristics.
The placement ofP. productus in group III
must notbeconsidereda reliableclassification
sinceonlyonestrain wasavailable foranalysis.
Thus,as seen inTable4, group
Ill
containsfiveorganisms and any two organisms considered
equivalent by any taxonomist were always
placedinthe same LCFAgroup.
On the basis of the LCFAprofiles ofwhole cells, asingle species, P. saccharolyticus, was
placed in group IV. P. saccharolyticus
con-tained
fatty
acids C14, C16,Ci8:1,
and C18 that appeared to be typical of the other peptococci and peptostreptococci.Thisorganism,however,had twounique characteristics not seen inthe
other peptococci and peptostreptococci: (i) sig-nificantamountsof
C20
wereproduced;and (ii)a prominent, unidentified peak with a
reten-tion time between C14 and
C16
was present.This peak may be a branched-chain 15-carbon
fatty acid similar to that reported in cell
ex-tractsofthe genus Ruminococcus (1). Some of
theliterature on the peptococci and
peptostrep-tococci support placing P. saccharolyticus in a
separate group. Dowell and Hawkins(5),inthe
Center for Disease ControlLaboratoryManual,
listed P. saccharolyticus astheonly speciesin
the peptococci. Rogosa (14) claimed thatP.
sac-charolyticus was frankly saccharoclastic and
should be excluded from thegenusPeptococcus,
without, however, mentioningwhere the
orga-nism should be placed. On the basis of LCFA
analysis, these studies agree with Rogosa (14,
15) and with Dowell and Hawkins (5), thatP.
saccharolyticus is significantly different from
the other peptococci andpeptostreptococci and
should beplacedinagroup by itself.
This study showed that LCFA analysis can
be used as an aid intaxonomy,but cannot be
relied on as a definitive test forspecies
designa-tion. Further work must be doneifthe
specia-tion of thepeptococci andthepeptostreptococci
is tobe clarified. Additional biochemical tests,
serological studies, guanine-cytosine ratios, or
deoxyribonucleic acid homologystudies might
prove helpful for speciation. The grouping of
speciesobtained from the LCFAprofilesmaybe
an indicationthat thesespeciesdooverlapeach
other agreat deal.Perhapsourgoalshould not
be to describenew species ofthese organisms,
but to consolidate the species known to exist.
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LCFAs OF PEPTOCOCCI AND PEPTOSTREPTOCOCCI 521
Thisapproachseems morelogical,atleast until
definitivepathogenicitystudieshave been
com-pleted.
ACKNOWLEDGMENTS
Thisstudywassupported by the Wisconsin Alumni
Re-search Federation. Wearegrateful forthecooperationof the members of the GeneralBacteriology Laboratoryatthe WisconsinStateLaboratory of Hygiene. Weespecially ap-preciateassistance givenby A. Helstad. We also wishto acknowledge E. Balish for his advice in thepreparation of this manuscript.
LITERATURE CITED
1. Allison, M. J., M. P. Bryant, I. Katz, and M. Ke_a_y. 1962. Metabolic function of branched-chain volatile fatty acids,growth factors for ruminococci. II.
Bio-synthesis of higher branched-chain fattyacids and aldehydes.J. Bacteriol. 83:1084-1093.
2. Amatein,C. F., and P. A. Hartman.1973. Differentia-tionofsomeenterococci bygaschromatography. J.
Bacteriol. 113:38-41.
3. Balows, A., R. M. DeHaan, V. R. Dowell, Jr., andL.B. Guze(ed.).1974.Anaerobic bacteria: role in disease. Charles C Thomas,Springfield, ill.
4. Dowell, V. R.1974.Anaerobic cocci.Newsletter of No-vember, 1974. Center for Disease Control, Atlanta, Ga.
5. Dowell, V. R., Jr., and T.M.Hawkins. 1974. Labora-torymethods inanaerobic bacteriology. CDC labora-torymanual. DHEWpublicationno.(CDC) 74-8272. U.S. Government Printing Office,Washington, D.C. 6. Holdeman, L. V. 1974. Anaerobe newsletter, no. 2. Anaerobe Laboratory, Virginia Polytechnic Institute andStateUniversity, Blacksburg, Va.
7. Holdeman, L. V., and W. E. C. Moore (ed.). 1973. Anaerobe laboratory manual,2nded.Anaerobe Lab-oratory, Virginia Polytechnic Institute and State
University,Blacksburg,Va.
8. Holdeman, L. V., and W. E. C. Moore. 1974. New genus, Coprococcus, twelve new species, and emended descriptions of four previously described speciesof bacteria fromhuman feces. Int. J. Syst. Bacteriol. 24:260-277.
9. Lambert, M. A., and C. W.Moss.1976. Cellular fatty acid composition ofStreptococcusmutansand related streptococci. J. Dent. Res. Special Issue A 55:A96-A102.
10. Mos,C.W., D. S.Kellogg, D. C. Farshy, M. A. Lam-bert, and J. D.Thayer. 1970.Cellularfatty acidsof pathogenic Neisseria. J. Bacteriol. 104:63-68. 11. Moss, C. W., M. A.Lambert, and W. H. Kerwin. 1974.
Comparison of rapidmethodsfor analysis ofbacterial fatty acids.Appl.Microbiol. 28:80-85.
2. Moss, C. W., S. B. Samuel, and R. E. Weaver. 1972. Cellularfattyacidcompositionofselected Pseudomo-nasspecies.Appl.Microbiol. 24:596-598.
13. Pien, F. D., R. L.Thompson, and W. J. Martin. 1972. Clinical and bacteriologicalstudiesof anaerobic gram positivecocci.MayoClin. Proc. 47:251-257. 14. Rogosa, M. 1974.Genus I. Peptococcus, p. 518-522. In R.
E. Buchananand N. E. Gibbons (ed.), Bergey's man-ual of determinativebacteriology, 8th ed. The Wil-liamsand Wilkins Co., Baltimore, Md.
15. Rogosa,M. 1974. GenusII.Peptostreptococcus,p. 522-525. In R. E. Buchannan and N. E. Gibbons (ed.),
Bergey'smanual ofdeterminative bacteriology, 8th ed. The Williams and Williams Co., Baltimore, Md. 16. Sutter, V. L., V. L. Vargo, and S. M. Finegold. 1975. Wadsworth anaerobic bacteriology manual, 2nd ed. Anaerobic BacteriologyLaboratory, Wadsworth Hos-pital Center, Los Angeles, Calif.
17. West, S. E. H.,and L. V. Holdeman. 1973. Placement of the namePeptococcus anaerobius (Hamm) Douglas onthe list of nominarejicienda.Int. J.Syst. Bacte-riol.23:283-289.
18. Zabranski, R. J. 1970. Isolation of anaerobic bacteria from clinical specimens. Mayo Clin. Proc. 45:256-264. VOL. 4, 1976