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0095-1137/84/010063-05$02.00/0

Copyright ©1984, American Societyfor Microbiology

Characterization

of

Neisseria cinerea,

a

Nonpathogenic Species

Isolated

on

Martin-Lewis Medium Selective for Pathogenic Neisseria

spp.

JOAN S.

KNAPP,'*

PATRICIA A. TOTTEN,1 MARTHA H. MULKS,2AND BARBARAH. MINSHEW3 Neisseria ReferenceLaboratory and Department of Medicine,

Univ'ersity

of Washington, Seattle, Washington 981951;

DepartmentofMedicine,

Tuifts-New

England Medical Center Hospital, Boston, Massachiusetts 021112; and Clinical

Microbiology Laboratory, Department of Pathology, SeattlePlublicHealth Hospital, and Department of

Laboratory

Medicine, University of Washington, Seattle, Washington

981143

Received 16 June 1983/Accepted 3 October 1983

An asaccharolytic, gram-negative, oxidase-positive diplococcus was isolated on Martin-Lewis medium

from thecervix of apatient attending an arthritisclinic at Seattle Public Health Hospital, Seattle, Wash.

Thisstrain,NRL32165, did not producedetectableacid from glucose, maltose, sucrose, fructose,mannitol,

orlactose in eithercystineTrypticase agar (BBL Microbiology Systems, Cockeysville, Md.) or modified

oxidation-fermentation medium and was identified presumptively as aglucose-negative Neisseria

gonor-rhoeaestrain,but wasidentifiedlater asNeisseriacinerea on the basis of itsbiochemical reactions. Nitrate

was notreduced,nitrite(0.001%,wt/vol) wasreduced,andpolysaccharidewas notproduced fromsucrose.

Proline, arginine,andcystine-cysteine wererequired for growth on defined medium. Strain NRL 32165 did

not react with antigonococcal protein I monoclonal antibodies and did not produce immunoglobulin A

protease.InDNA:DNA homology studieswith N.gonorrhoeae NRL8038(F62) andN.cinerea type strain

NRL30003, strain NRL 32165 showed95%homologyrelative to N. cinereaand 44% homology relative to

N. gonorrhoeae. Thus, the identity of strain NRL 32165 was confirmed as N. cinerea (von Lingelsheim

1906)Murray 1939. Of allNeisseriaspp., N. cinerea is most likely to bemisidentifiedas aglucose-negative

N. gonorrhoeae strain.

Commensal

colistin-susceptible

Neisseria spp. and

Bran-hamellacatarrhalis areonlyrarelyisolatedon media

selec-tive for the pathogenic Neisseria spp., Neisseria

gonor-rhoeae andNeisseria meningitidis, andaredifferentiatedon

the basis of their cultural characteristics, distinctive sugar

utilization reactions, ability toreduce nitrate,and ability to

produce

polysaccharide

fromsucrose. Accordingto current

literature(23, 33), among commensal species isolated from

humans, only Neisseria flavescens and B. catarrhalis are

asaccharolytic.

Another asaccharolytic Neisseria sp., Neisseria cinerea,

was first described by von Lingelsheim in 1906 (35) and

namedMicrococcus

cinereius.

Itwasdescribed subsequently

as Neisseria

pseudocatarrhalis

(F. M. Huntoon, Abstr. Annu.Meet. Am. Soc.Bacteriol. 1934,M50, p. 108) but was

assigned

to the species N. cinerea (27) since the

epithet

cinereahad

priority

taxonomically. Since then,manystrains appear to have been isolated butincorrectly identifiedasB.

catarrhalis(8, 11). Thesespecies differ biochemically onlyin

their ability to reduce nitrate, a test which was not

intro-ducedintotheclassification ofNeisseria spp. until1961(1).

B. catarrhalis strains reduce nitrate, whereas N. cinerea

strains do not. Bergerand Paepcke (3) "rediscovered" and

describedN. cinerea in 1962 andshowed thattherewas no

antigenic relatedness between N. cinerea and Neisseria

catarrhalis;the latter

species

was

subsequently reassigned

to

the genus Branhamella (5) in 1970. N. cinerea was not

described inBergey's Manual

of

Determinative

Bacteriolo-gy, 8thed. (33)ortheManual

of

Clinical

Microbiology,

3rd

ed.(23),butit will bedescribedin the9th edition of

Bergey's

Manual(N.A. Vedros,

personal

communication).

Early studies indicated thatN. cinerea strains colonized

the nasopharynx (2, 8, 11). Commensal Neisseria spp. and

*Correspondingauthor.

B. catarrhalis were isolated only rarely fromthe

genitouri-nary tract (12, 14, 36, 38). The prevalence ofN. cinereais

uncertain because early investigators identified N. cinerea

strains as a subtype ofB. catarrhalis (8, 11). Berger and

Paepcke (3) foundthat N. cinereastrains accounted forca.

93% of 27 asaccharolytic Neisseria

(Branhamella)

spp.

isolated from the nasopharynx of healthy subjects. N.

cin-erea is regarded as a nonpathogenic species, although it

appears to have been isolated from the genitourinary tract

(14, 36) and from the cerebrospinal fluid of a

patient

with acute meningitis (31). In some cases, the isolates were

identifiedas N.

flavescens,

N.

gonorrhoeae,

orB.

catarrha-lis, butreevaluation ofthese

publications

(2) indicates that

they wereprobably N. cinerea strains.

Inthisstudy,wedescribetheisolationon

colistin-contain-ing Martin-Lewis medium of a

colistin-susceptible,

asac-charolytic,

gram-negative, oxidase-positive diplococcus

whose biochemical characteristics did not conform to the

description

of either N.

flavescens

or B.

catarrhalis,

but whosecultural

characteristics

resembled those ofN.

gonor-rhoeae. This strain was

subsequently

identified as N.

cin-erea(von

Lingelsheim

1906) Murray1939

(27).

Weevaluated

biochemicaltestsandreference

techniques

whichwillaidin

identifying N. cinerea strains and

differentiating

them from

N.

gonorrhoeae.

MATERIALSAND METHODS

Isolation andmaintenance. An

asaccharolytic,

gram-nega-tive, oxidase-positive diplococcus was isolated on

Martin-Lewis medium (Prepared Media Laboratory,

Tualatin,

Oreg.) fromthecervix ofa

19-year-old

woman

subsequently

diagnosedashaving

psoriatic

arthritis. The strain,

designat-ed NRL 32165, wassubcultured onchocolateagarmedium.

Sincereceiptin theNeisseriaReference

Laboratory (NRL),

63

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TABLE 1. Summary ofdifferential characteristics for N. gonorrhoeae, asaccharolyticNeisseria spp.,and B.catarrhalisa

Acid from: Polysac- Require-

Produc-Species G M S F Ma Nitrate charide ment for tion of

Crlistin

gron

h

Species G M S F Ma L reduction fromsu- cystine- IgApro- reit onM

crose cysteine tease ance agar

N. gonorrhoeae + - - - + + +

N. cinerea - - - + - - +

N.flavescens - - - + + - - +

B.catarrhalis - - - + - - - - +

'Abbreviations:G, glucose; M,maltose;S,sucrose;F,fructose; Ma,mannitol; L,lactose;MH,Mueller-Hinton. +,Positiveforallstrains

tested; -, negativefor all strains tested. DataforN.gonorrhoeaearebasedonliterature citations(2,6, 23, 33).

the strain has been cultured on GC base medium (Difco

Laboratories, Detroit,

Mich.) plus

defined supplement

(GCK) (37)at36°CinaCO2-enriched

atmosphere.

The strain

was stored in50%gammaglobulin-freehorse serum (GIBCO

Laboratories, Grand Island, N.Y.) in tryptic soy broth

(Difco) at -70°C.

Strains. N. cinerea type strain NRL 30003 (ATCC 14685)

and N.

gonorrhoeae

NRL 8038

(F62)

were used asreference

strains for DNA:DNA homology studies. Species used in

comparative studies to determine differential biochemical

characteristics arelisted in Table 1.Two N. cinerea strains

(NRL 32824andNRL32828)isolatedonsheep bloodagar in Seattle in 1981 werealsostudied. Inaddition, 10strains ofB. catarrhalis and the taxonomic type strain ofN.flavescens

(NRL 30009) were studied.

Colonymorphology. Observations of the colonial

morphol-ogy of strains were made on GCK medium, human and sheep blood agar, and chocolateagar. Strains were streaked

onto each medium and colony

morphology

was described

after incubation for 24 h at 36°C in a CO--enriched atmo-sphere.

Sugar utilization tests. Strains were tested in cystine

Trypticase agarmedium(BBLMicrobiology Systems,

Cock-eysville, Md.) for theirabilitytoproduce acidfromglucose,

maltose, lactose, and sucrose. Subsequent

confirmatory

tests were made inmodifiedoxidation-fermentationmedium

containingglucose, maltose, sucrose,fructose, mannitol, or

lactose (19). Modified oxidation-fermentation base medium

has thefollowing composition (per liter): 0.2% Difco

prote-asepeptone no. 3-0.5% NaCI-0.03%dipotassium hydrogen

phosphate-0.3%agar-0.5 ml of a0.17%phenol red solution.

Additional biochemical tests. Tests to detect nitrate and

nitrite reduction and polysaccharide production from

su-crose wereperformed as described previously (3, 19). Auxotyping. Strains were tested on chemically defined

medium (NEDA) as described previously (18).

Require-ments for proline, arginine, hypoxanthine, uracil,

methio-nine,andcystine-cysteine were determined. Type strains of

asaccharolytic species N. flavescens and B.

catarrhalis

were

compared with N. cinerea strains.

Antibioticsusceptibility tests. Susceptibility ofN. cinerea strains topenicillin,tetracycline, spectinomycin,

erythromy-cin, and vancomycin were determined as described previ-ously (18). Colistin susceptibility was determined by disk diffusion tests on GCK agarwith BBL disks (10

jig).

Serology. Serogrouping ofgonococciwasperformed using

thecoagglutination technique. Reagent staphylococci

(pro-vided by Lars Rudin, Pharmacia AB, Uppsala, Sweden)

were sensitized with each of 10 monoclonal antibodies

(provided by Milton Tam, Genetic Systems Corp., Seattle,

Wash.) against N. gonorrhoeae (34). Each test was

per-formed by mixing one drop of a suspension of boiled

organism

with one drop of a suspension of the reagent

staphylococci,rotating for 2min,andobserving the mixture

under oblique transmitted light for agglutination. The re-agents used were chosen to subgroup gonococci into sero-groups WI, WII, and Wlll, but cumulatively they may be used to identify gonococci since cross-reactions with other Neisseria spp. have not been observed (29).

Detectionof IgA protease.Immunoglobulin A(IgA) prote-ase was detected using sodium dodecyl

sulfate-polyacryl-amidegel electrophoresisas describedpreviously (20,25).

Isolationof DNA. DNA was isolated bythe procedure of Brenneretal. (4), and the final concentration was calculated from 260 to 280 nm absorption in a Gilford model 2400 spectrophotometer (Gilford Instrument Laboratories, Inc.,

Oberlin, Ohio).

Preparation of radiolabeled DNA. For hybridization ex-periments, whole-cell DNA from strain NRL 32165 was labeled to a specific activity of 2 x 106cpm/Lgof DNA by

the nick translation technique of Maniatis et al. (21) with

[3H]deoxyribosylthymine triphosphate (77 Ci/mM; New

En-gland Nuclear Corp., Boston, Mass.). Nick translation was performed with the DNase concentration adjusted to yield

DNA with a final fragment size of 3,000 base pairs, after which the DNA was sonicated to500 base pairs.

Whole-cell DNA:DNA homology studies. Whole-cell DNA was hybridized overnight, and homology was analyzed by thesingle-strand endonuclease procedure of Crosa et al. (7) asmodifiedby Piotet al. (30).Homologieswere determined afterhybridization at 65°C overnight in 0.42 MNaCl, which were optimal conditions for hybridization, i.e., 25 to 30°C

belowthemelting point ofthe probe DNA.

RESULTS

An asaccharolytic, gram-negative, oxidase-positive

diplo-coccus was isolated on Martin-Lewis medium from the cervix ofa patient attendingan arthritis clinic. This strain, NRL32165, was tested in the clinical laboratory for its sugar

utilization pattern in cystine Trypticase agar and failed to

produce acid fromglucose,maltose, sucrose, or lactose. The strain was also resistant to erythromycin by disk diffusion

testingand was studied further.

Colonialmorphology. N. cinerea colonies on GCK

medi-umincubatedat36°C for 24 h weregold-brown;the cell mass

was

often more

distinctly

gold-brown than N.

gonorrhoeae.

Colonies were ca. 1 mm in diameter with a glistening

surface, entire margin, and convex elevation (similar to T3

coloniesofN.gonorrhoeae) and weretranslucent and easily

emulsified in broth. Cellsshowed the diplococcal morpholo-gy, in whichcells hadadjacentsides flattened and varied in

size. Pairs or clumps predominated over individual

diplo-cocci. Giant cells were sometimesseenin 48-hcultures(2).

Coloniesonsheepand humanbloodagar and chocolateagar

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were grey-white. Cell and colonial morphology conformed with the description of N. cinerea (3).

Biochemical characteristics. The biochemical characteris-tics of strains NRL 32165, NRL 32824, and NRL 32828 agree with those reported previously for N. cinerea (3). Sugar utilization tests performed in both cystine Trypticase agar medium and modified oxidation-fermentation medium con-taining glucose, maltose, sucrose, lactose, mannitol, or fructose confirmed that the strains did not produce acid from any of these carbohydrates even after incubation for 14 days. Strains did not reduce nitrate or produce polysaccharide from sucrose, but they did reduce nitrite (0.001%, wt/vol),as did N. gonorrhoeae strains (J. S. Knapp, Abstr. Annu. Meet. Am. Soc. Microbiol. 1983, D27, p. 63). On the basis of biochemical tests, the strains were identified as N. cinerea.

Nutritional requirements. Determination of the require-ment of the strains for cysteine-cystine was made after incubation for 48 h at

36°C

in a CO2-enriched atmosphere. All N. cinerea strains required cysteine-cystine for growth on NEDA medium. In addition, all strains also required proline and arginine (Pro- Arg-). N. flavescens also re-quired cysteine-cystine for growth, but B. catarrhalis did not.

Serology. N. cinerea, B. catarrhalis, and N. flavescens strains did not react with any of 10 antigonococcal protein I monoclonal antibodies in coagglutination tests.

Antibiotic susceptibility patterns. N. cinerea strains were susceptible to various antibiotics measured in minimal in-hibitory concentrations (micrograms per milliliter) as fol-lows. Penicillin (0.125 to 1.0), tetracycline (0.125 to 0.5), vancomycin (32 to .64), erythromycin (2.0 to 4.0), and spectinomycin (16 to .32). The results demonstrated that antibiotic susceptibility patterns for N. cinerea strains were not notably different from those for N.gonorrhoeae. Resist-ance to erythromycin was higher than normally found forN. gonorrhoeae strains (13, 28). Colistin disk susceptibility

tests with N. cinerea strains showed that zonesofinhibition were :10 mm in diameter with anunstandardized inoculum.

In contrast, N. gonorrhoeae strains showed no inhibition. These results are consistent with those reported by Berger (2).

IgA protease activity. None of the N. cinerea strains produced IgA protease. Strains of N. gonorrhoeae and N. meningitidis produce IgA proteases, whereas commensal Neisseria spp. do not (26). In accordance with these find-ings, the strains of N. cinerea examined in this study failed to produce IgA protease, thus providing anadditional refer-ence test which can aid in their identification.

DNA:DNA homologystudies. Homology studieswere per-formed using 3H-labeled whole-cell DNA from N. cinerea

NRL 32165 and unlabeled DNA from the test organisms.

The actual extent of reassociation of homologous (strain NRL 32165) and heterologous (calfthymus) DNA averaged

75 and 7%, respectively. Although thetypestrain N. cinerea

NRL 30003 was tested, we did not use it as the reference

strain for these studies because it contained a small, 2.0-megadalton (Mdal) plasmid. Since most small plasmids oc-cur in multiple copies (often 20 to 30) per cell, we were concerned that the presence ofthisplasmidmight affect the homology results measurably. Results ofhomology studies are summarized below; each value istheaverageof four or five determinations. The type strain NRL30003 and strain NRL 32824 showed95and 94%relative homology, respec-tively, with strain NRL 32165; strain NRL 32828 showed

81% homology relative to strain NRL32165. The

compara-tively low value of 81% relative homology between strains

NRL 32828and NRL 32165compared withhigher values for

theother strains might be accounted for inpart

by

the

3.2-Mdal plasmidcarried by strain NRL 32828.If thisplasmidis presentin 20 to 30copies percell, it wouldaccountforca.

6% ofthe DNA in this strain (22). In contrast, N.

gonor-rhoeae NRL 8038 showed only 44%

homology

relative to strain NRL 32165. These studies confirm that strain NRL 32165 and other recently isolated strains were N. cinerea

(vonLingelsheim 1906) Murray 1939.

DISCUSSION

A strain of N. cinerea (von

Lingelsheim 1906)

Murray

1939 wasisolatedfromaclinical

specimen

on Martin-Lewis

medium selective for pathogenic Neisseria spp. This

isola-tion is noteworthy since N. cinerea is not

currently

listed

among human Neisseria spp. (23,

33).

N. cinerea has not

been correctly identified

previously

in the United

States,

although it appears that strains of this

species

have been

isolated from clinical

specimens

in the United States and

Europe on severaloccasions but

incorrectly

identifiedasB. catarrhalis (14),N.flavescens (31),N.

gonorrhoeae

(32),

or

N. pseudocatarrhalis (Huntoon, Abstr. Annu. Meet. Am.

Soc. Bacteriol. 1934).

Previoushomology studieshave shownthat strains

classi-fiedin thesame

species

show70%or morerelative homolo-gyunderoptimal conditions(4). DNA:DNA

homology

stud-ies by different methods have

reported

different relative

homology values for

interspecific

reactionsbetween strains

representative ofNeisseria spp. Hoke andVedros

(15)

used

the thermal renaturation

technique

and

reported

60%

rela-tive homology between N.

gonorrhoeae

and N.

cinerea,

Neisseria sicca, Neisseria

subflava,

and N.

flavescens.

In contrast, Elwell and Falkow

(9),

using

the more restrictive S1 endonuclease assay, demonstrated relative

homology

values for interspecific reactions between the DNA of N. gonorrhoeae, N. sicca, N.

subflava,

and N.

flavescens

ranging between 24 and

29%;

N. cinerea strains were not

tested in that

study.

Although

no DNA:DNA

homology

studiesby theS1 endonuclease

procedure

have been

report-edpreviously betweenN.

gonorrhoeae

and N.

cinerea,

the

relative homology of 44% between these

species

suggests

that N. cinerea may be more

closely

related to N. gonor-rhoeae than are other commensal Neisseria spp. Evidence

that N. cinerea is more

closely

relatedtoN.

gonorrhoeae

in

fatty acid

composition

rather than to the other commensal

Neisseria spp. and B. catarrhalis has been

reported

previ-ously (16, 17).

Theisolation ofN. cinerea

emphasized

weaknesses inone

reference test used to confirm the identification of

gono-cocci. Catlin(6) noted that

although

all strains ofN.

gonor-rhoeaerequired

cystine-cysteine

for

growth

onNEDA

medi-um, more than90% ofN.

meningitidis

strains did not, and

some N. lactamica strains

required

cystine-cysteine,

where-asothers didnot. Ourobservationsin

auxotyping

N.

gonor-rhoeae,N.

meningitidis,

andN. lactamica strains since 1973

haveconfirmed thoseofCatlin. In

addition,

in this

study

we

found thatN. cinerea andN.

flavescens

strains

required

not only

cystine-cysteine

for

growth

on NEDA

medium,

but N. cinerea strains also

required

proline

and

arginine

(Pro-Arg-)forgrowth,thus

contributing

furthertoconfusion with

gonococci.

Consequently,

the

requirement

for

cystine-cyste-ine cannot be used to differentiate between N. cinerea and

N. gonorrhoeae.

Although in

coagglutination

tests all

gonococci

reacted

with at least 1 of 10 research

antigonococcal

protein

I monoclonal antibodies

(29, 34),

no cross-reactions were

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observed with other Neisser-ia spp., including N. (inerea strains and B. (at(lrrhalis. However, because commercial serological diagnostictestsforN.

gonorrhoeae

donot detect allstrains (10;J. P. Libonati,R. L. Leilich, and L. Loomis. Abstr. Annu. Meet. Am. Soc. Microbiol. 1983, C19, p.314), commercial diagnostic reagents are oflimited usefulness for distinguishing N.

gon0orrh-loeae

strains from N. (inerei strains.

Additional tests which proved useful in differentiating N. (cinerei(l from N. gono-rrhoeie were the ability of N. cinerea

strains to grow on simple media such as tryptic soy agar (Prepared Media Laboratory)orMueller-Hintonagar(Difco) and their susceptibility to colistin. Tryptic soy agar and Mueller-Hinton agar supported good growth of N. cinerea strains but not N. gonor-hoeae strains. N. (inerea strains were susceptible tocolistin (10-pLg disk; BBL) but resistant

toerythromycin and of intermediate resistance topenicillin. Although N. cinerea appears to be an extremely rare isolate fromgenital sites (12, 14, 36, 38), it has been isolated more frequently from the nasopharynx (3) and oropharynx (J. S. Knapp, P.A.Totten, B. H. Minshew, andE. W. Hook

III, Abstr. Annu. Meet. Am. Soc. Microbiol. 1983, C26, p. 316). The presence of N. cinerca in these sites poses two

areasofpotential confusion for clinical microbiologists iden-tifyingNeisseriaspp.First,since N. cinerea coloniesclosely

resemble N. gonorrhoeae colonies, these species may be confusedwhenacid productionfromglucose is weak. Thus, N. cinerea strains may be misidentified as N. gonorrhoeae when isolated on selective media oron nonselective media often used nowto detect vancomycin-susceptible

gonococ-cal strains. Second,N. cinerea strainsmaybeconfusedwith

B. catarrhalis strains when isolated from pharyngeal speci-mens onnonselective media employedtoisolate Haemophi-lus influenzae. Since B. catarrhalis is being increasingly implicatedas arespiratory pathogen, it is important that the

etiological agents in theseinfections be identified correctly. AlthoughN. cinereacolony morphology is distinctfrom that

ofB.catarrhalis strains,N. ciner-eastrainswereconsistently misidentified as aB. catarrhalis subtype (8, 11) before the

introduction of nitrate reduction as a differential test in Neisseria spp. classification in 1961 (1). Confusion of N. (ine)(ea withN.

flai'escens

(31) resulted because the strains exhibit similar colony morphologies and also because pro-duction ofpolysaccharide from sucrose was not introduced into Neisseria spp. classification until 1961.

SinceN. (cinerea isnotlistedamonghumanNeisseriaspp.

incurrentreference texts, asummary of differential

charac-teristics useful for differentiating between asaccharolytic species of N. (inerea, N. flaves(cens, B. caitarrhalis, and weakacid-producing N. gonorrhoeae strains is provided in Table 1. Unfortunately, DNA:DNA homology studies

pro-videthe onlyconfirmatorytestavailablecurrentlytoidentify N. cinerea. Tests used in clinical laboratories permit the

identification ofN. cinerea onlyby the elimination ofother

currently recognized Neisseria spp. or B. catarrhalis. In

viewof thelegal and scientific implications of misidentifying N. (cinerea strainsas N. gonorrhoeae orB. catallhalis, we

wish toalert clinical microbiologists to the existence ofN.

(inerea and to advise caution when identifying apparently

glucose-negative, gram-negative, oxidase-positive

diplo-cocci.

ACKNOWLEDGMENT

This researchwas supported by Public Health Servicegrant

Al-12191fromthe National Institutes of Health.

LITERATURE CITED

1. Berger, U. 1961. Reduktion vonnitrat andnitritdurchNeisseria. Z. Hyg. 148:45-50.

2. Berger, U. 1963. Dieanspruchslosen Neisserien. Ergeb. Mikro-biol. Immunitaetsforsch. 36:97-167.

3. Berger, U., and E. Paepcke. 1962. Untersuchungen uber die asaccharolytischen Neisselriendesmenschlichen Nasopharynx. Z. Hyg. 148:269-281.

4. Brenner, D. J., A. C. McWhorter,J. K. L. Knutson, and A. G. Steigerwalt. 1982.Escherichiaiulnaris: a newspeciesof Elitero-bacteriaceae associated with human wounds. J. Clin. Micro-biol. 15:1133-1140.

5. Catlin, B. W. 1970. Transfer ofthe organism named Neisseria c-atatrrhhalis to Brcanhainella gen. nov. Int. J. Syst. Bacteriol. 20:155-159.

6. Catlin, B. W. 1973. Nutritional profiles of Neisseriia

gonor-rhoeae, Neisserii ineningitidis, Neisseria Iactainica in chemi-cally defined media and the use of growth requirements for gonococcal typing. J. Infect. Dis. 128:178-194.

7. Crosa, J. H., D. J. Brenner, and S. Falkow. 1973. Use of single-strand specific nuclease for analysis of bacterial plasmid deoxy-ribonucleic acid homo- and heteroduplexes. J. Bacteriol. 115:904-911.

8. Elser, W. J., and F. M. Huntoon. 1909. Studies onmeningitis. J. Med. Res. 20:371-541.

9. Elwell, L. P., and S. Falkow. 1977. Plasmids of the genus Neisseria, p. 127-154. In R. B. Roberts (ed.). The gonococcus. John Wiley & Sons, Inc.. New York.

10. Futrovsky, S. L., C. A.Gaydos, and J. Keiser. 1981. Comparison of the Phadebact gonococcus test with the rapid fermentation method. J. Clin. Microbiol. 14:89-93.

11. Gordon, J. E. 1921. The gram-negative cocci in "colds" and influenza. VII. Influenza studies. J. Infect. Dis. 29:462-494. 12. Gurd, F. B. 1908. A contribution to the bacteriology of the

female genital tract with special reference to the detection of the gonococcus. J. Med. Res. 18:291-303.

13. Hall, W. H., E. A. Schierl,and J. E. Maccani. 1979. Compara-tive susceptibility ofpenicillinase-positive and -negative Neis-seria gonorrhoeie to 30 antibiotics. Antimicrob. Agents Che-mother. 15:562-567.

14. Henriksen, S. D. 1946. Isolation of atypical strains ofNeisselia catarrhalisfromthe genito-urinary tract. Acta Derm. Venereol. (Stockholm) 26:506-514.

15. Hoke, C., and N. A. Vedros. 1982. Taxonomy of the Neisseriae: deoxyribonucleic acid base composition,interspecific transfor-mation. and deoxyribonucleic acidhybridization. lnt. J. Syst. Bacteriol. 32:57-66.

16. Jantzen, E., K. Bryn, T. Bergan, and K.

Bovre.

1974. Gas

chromatography ofbacterial whole cell methanolysates. IV. A procedure for fractionation and identification of fatty acids monosaccharides ofcellular structures. Acta Pathol. Microbiol. Scand. Sect. B 82:753-766.

17. Jantzen, E., K. Bryn, T. Bergan, and K.

Bovre.

1974. Gas chromatography of bacterial whole cell methanolysates. V. Fatty acid composition of neisseria and moraxellae. Acta Pathol. Microbiol. Scand. Sect. B 82:767-779.

18. Knapp, J. S., and K. K. Holmes. 1975.Disseminatedgonococcal infections caused by Neisseria

gonorrhloeae

with unique nutri-tional requirements. J. Infect. Dis. 132:204-208.

19. Knapp, J. S., and K. K. Holmes. 1983. A modified oxidation-fermentation medium for detection of acid production from carbohydrates byNeisserial spp. andBrain/wilcnellacatarrhalis. J. Clin. Microbiol. 18:56-62.

20. Laemmli, U. K. 1970. Cleavage of structural protein during the assembly of the head of the bacteriophage T4. Nature (London) 227:680-685.

21. Maniatis, T., A. Jeffery, and A. G. Kleid. 1975. Nucleotide sequence of the rightward operator of phage lambda. Proc. NatI. Acad. Sci. U.S.A. 72:1184-1188.

22. Mayer, L. D., K. K. Holmes, and S. Falkow. 1974. Characteriza-tion of plasmid deoxyribonucleic acid from Neisseriia

gonor-rholecae. Infect. Immun. 10:712-717.

on April 10, 2020 by guest

http://jcm.asm.org/

(5)

23. Morello, J. A., and M. Bohnnoff. 1980. Neisseria and Branh/a-mella,p.111-130. In E. H.Lennette,A.Balows,W.J. Hausler,

Jr.,and J. P. Truant(ed.),Manual of clinicalmicrobiology,3rd ed.American Society for Microbiology, Washington, D.C. 24. Morello, J. A.,S. A. Lerner, and M. Bohnhoff. 1976.

Character-istics of atypical Neisseria gonorrhoeae from disseminated and localized infections. Infect. Immun. 13:1510-1516.

25. Mulks, M. H., S. J. Kornfeld, andA. G. Plaut. 1980. Specific proteolysis of human IgA by Streptococclus pneiurnoniiae and Haemnopli/us infliuenzae. J. Infect. Dis. 141:450-456.

26. Mulks, M. H., and A. G. Plaut. 1978. IgAproteaseproductionas

a characteristic distinguishing pathogenic from harmless

Neis-seriaceae. N. Engl. J. Med. 299:973-976.

27. Murray, E. G. D. 1939. Genus I. Neisseria Trevisan, p.

278-288. In D. H.Bergey, R.S. Breed, E. G.D.Murray,and A. P.

Hitchins (ed.). Bergey's manual of determinative bacteriology. 5th ed. The Williams & WilkinsCo.. Baltimore.

28. Ng, W. S., P. Anton, and K. Arnold. 1981. Neisserial

gonor-r-lioeae strains isolatedin Hong Kong: invitrosusceptibility to

13antibiotics. Antimicrob. Agents Chemother. 19:12-17. 29. Nowinski, R. C., M. R. Tam, L. C. Goldstein, L. Stong, C.-C.

Kuo, L. Corey, W. E. Stamm, H. H. Handsfield, J. S. Knapp, and K. K. Holmes. 1983. Monoclonal antibodiesfordiagnosis of infectious diseases in humans. Science 219:637-644.

30. Piot, P., E. Van Dyck,M. Goodfellow, and S. Falkow. 1980. A

taxonomic study of Gardnerella ivaginalis (Haetnophillus iagi-na/lis) Gardner & Dukes 1955. J.Gen. Microbiol. 119:373-396. 31. Prentice, A. W. 1957. Neisseria flai'escens as a cause of

meningitis. Lancet i:613-614.

32. Reyn, A. 1948. Atypical gonococcal strains. Acta Derm. Vener-eol. (Stockholm) 28:381-386.

33. Reyn, A. 1974. Genus 1. Neisseria Trevisan, p. 428-432. In R. E. Buchanan and N. E. Gibbons(ed.), Bergey's manual of determinative bacteriology, 8th ed. The Williams & Wilkins Co.. Baltimore.

34. Tam,M.R., T. M. Buchanan, E. G. Sandstrom, K. K. Holmes, J. S.Knapp, A. W. Siadak, and R. C. Nowinski. 1982. Serologi-cal classification of Neisseriia gonorrhoeae with monoclonal antibodies. Infect. Immun. 36:1042-1053.

35. Von Lingelsheim, W. 1906.Die bakteriologischen Arbeitender Kgl Hygienischen Station zu Beuthen 0. Schl. wahrend der Genickstarreepidemie in Oberschlesien im Winter 1904/05. Klin. Jahrb. 15:373-489.

36. Wax, L. 1950.Theidentity ofNeisseriiaother than the gonococ-cusfrom thegenito-urinary tract.J. Vener. Dis. Inf. 31:208. 37. White, L. A., and D. S. Kellogg, Jr. 1965. Neisseria

gonor-rhloeae identification in directsmearsbyafluorescent antibody-counterstain method. Appl. Microbiol. 13:171-174.

38. Wilkinson, A. E. 1952. Occurrence of Neisseria other thanthe gonococcus in thegenital tract. Br. J. Vener. Dis. 28:24-27.

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http://jcm.asm.org/

References

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