Phage Typing Set for Group C1 and C2Salmonellae

JOURNALOFCLINICAL MICROBIOLOGY, Feb. 1976,p.214-217 Copyright ©)1976 American Societyfor Microbiology

Vol. 3, No. 2 Printed inUSA.

Phage Typing Set for

Group

C1

and

C2

Salmonellae

M. GERSHMAN

DepartmentsofMicrobiology and Animal and Veterinary Sciences, University ofMaine, Orono, Maine 04473

Received forpublication 26 September 1975

Fifteencommonserotypesrepresentative ofgroupCl and C2 Salmonellawere

characterized usingasinglesetof phages.

Salmonellosis isawidespread disease

affect-ing man, livestock, and companion animals.

The incidence of Salmonella

infections

is inan

ascendancy and the ubiquitous natureof these

organisms appears toemphasize the futility of

total eradication. Nevertheless, effective

con-trolmeasures canbeapplied through the early

recognition ofcases.

The multitude of

Salmonella

serotypesin

ex-istence isnotparticularly disturbing for

diver-sitycanbe, andfrequently is,epidemiologically

useful. Many apparently unrelated cases have

beenlinked throughsomeuniquecharacteristic

possessed by anisolate.

Serological delineations are advantageous

butmaybe inadequate for

epidemiological

pur-poses. Commonplace serotypes, for example,

are difficult, ifnot impossible, to relate to a

particular outbreak unless measures are first

used to characterize the serotype involved.

Amongthe methods available for

differentiat-ing

Salmonella,

phage typing has

emerged

as a

technique providing both reliability and

maxi-mumstraindifferentiation(1).

Craigie and Yen (4) establishedaphage

typ-ingscheme for characterizing

Salmonella typhi

(4, 5). Its success led to the development ofa

numberof

schemes

for other

Salmonella

sero-types.They include S.

paratyphi

A(2), S.

para-typhi

B(1),

S. typhimurium

(3, 8, 14), S.

braen-derup (12), S.

blockley

(13), S. dublin and S.

enteritidis (9), S.

gallinarium,

andS.

pullorum

(10).

Inkeeping withour ownimmediate interests

and circumstances, phage typing sets for S.

thompson

(6)

and S.newport(7,

11)

were

devel-oped in our laboratory. In the course ofour

research itwasobserved that these phagesets

could be used todistinguishanumber ofother

Salmonellaserotypesalso

representative

of

se-rological groupsC and C2

Using

acombinedsetofS. thompson and S.

newportphageswewereabletotype 397ofthe

400 cultures examined at random. The lytic

pattern of thesephagesisnotedinTable 1.

Isolates to be typed were lightly inoculated

into 3 ml of nutrient broth and incubated at

37 C for 1.5 h or until turbidity was barely TABLE 1. Lytic pattern of typing phages at routine testdilutionsa

Salmonellathompson phages Salmonella newportphages

Typestrains - _

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

S.thompson 1 CL - CL - - _ _ _ _ CL CL +++ - OL CL

S. thompson 2 - CL - - <SCL ++ - CL _ - - SCL - OL

-S. thompson 3 +++ <SCL CL - ± <SCL CL OL - ++ OL - - OL

-S. thompson4 +++ - +++ CL _- <SCL ++± +++ ++ - CL

S. thompson 5 ++ ++ CL - CL CL CL OL - ++ OL ++ _ OL ++

S. thompson6 <SCL - CL - SCL CL SCL OL - - OL <SCL

-S.thompson 7 +++ CL CL - CL CL CL CL - CL CL - _ _ OL

S.thompson 8 _ - - SCL +++ SCL CL - _ _ _

S.newport 9 CL - SCL- _- - CL CL CL CL CL SCL CL

S. newport 10 _ - ++ . . _ CL SCL - +-++ CL

S. newport11 CL CL

+±-+-

- - - CL CL SCL - - CL

S.newport 12 SCL - SCL - - _ _ _ SCL SCL - CI. CL OL CL

S. newport13 SCL - _ _ _ _

_-

SCL - CL CL OL CL

S.newport 14 - _ - ++ CL OL

S. newport 15 +++

ICLI

- CL SCL SCL - OL CL

aAbbreviations: CL,

confluent

lysis; OL,opague lysis(opacitydue tosecondary

growth);

SCL, semicon-fluent lysis;<SCL, less than semiconfluent lysis; + + +, 120 plaques; + + ±, 81 to 120 plaques; ++, 61to80

plaques; +±, 41 to 60plaques; +, 21 to 40 plaques; +, 6 to 20plaques; -, 0 to 5 plaques.

214

on February 7, 2020 by guest

http://jcm.asm.org/

Downloaded from

on February 7, 2020 by guest

http://jcm.asm.org/

Downloaded from

on February 7, 2020 by guest

http://jcm.asm.org/

TABLE 2. Reactions of test strains atroutine testdilutionsa

Salmonella groupC, Culture

Patterns

Salmonella groupC, Culture Patterns

type strains no. typestrains no.

S. bareilly S.bareilly S. bareilly S. bareillyb S.bareillyb S.bareillyb S. bareillyb S. bareillyb S. bareilly S. braenderup S. braenderup S. braenderup S. cholerae-Suis var. kunzen-dorf S.eimsbuettel S. eimsbuettel S.eimsbuettel S. eimsbuettel S. eimsbuettel S. eimsbuettel S. eimsbuettel S. eimsbuettel S. eimsbuettel S. eimsbuettel S.eimsbuettel S. eimsbuettel S.eimsbuettel S. eimsbuettel S.infantis S. infantis S.infantis S. infantis S.infantis S.infantis S. infantis S. infantis S. infantis S. infantis S. infantis S. infantis S. infantis S.infantis S. montevideo S. montevideo S. montevideo S. montevideo S. montevideo S. montevideo S. montevideo S. montevideo S. montevideo S. montevideo S.montevideo S. montevideo S. montevideo S. montevideo S. montevideo 1 3/11 2 2/3 3 3 4 2/3/9/10/11 5 2/3/9/10/11 6 2/3/9/10/11 7 2/3/9/10/11 8 2/3/9/10/11 9 2/3/11 1 3 2 10/15 3 14 1 14 1 2/3/8/11/14 2 1/3/10 3 1/3/10/11/14/15 4 2/3 5 3/10/14 6 3/11/15 7 3 8 2/3/11 9 3/11/14 10 3/14 11 2/3/11/14 12 14 13 1/3/10/11/15 14 3/11 1 1/2/10/12/14/15 2 1/3/14 3 1 4 3/11/12/14 5 12/14 6 1/2/3/12/14 7 3/12/14 8 3 9 1/3/10/11/14/15 10 1/2/12/14/15 11 1/4 12 1/10/14/15 13 14 14 3/14 1 14 2 2/12/14 3 2/14 4 12/14 5 3/10/11/12/14/15 6 11/15 7 1/3/11/14 8 1/2/3/14 9 3/14 10 2/3/8 11 3/12/14 12 2/3/8 13 2/8 14 3 15 8/14 S. montevideo S. montevideo S. montevideo S. montevideo S. montevideo S. montevideo S. montevideo S. montevideo S.oranienburg S.oranienburg S.oranienburg S. oranienburg S.oranienburg S.oranienburg S. tennessee S. tennessee S. tennessee S. tennessee S. tennessee S. tennessee S.tennessee S. thompson S. thompson S. thompson S. thompson S. thompsonr S.thompsonr S. thompsonc S. thompsone S. thompsonc S. thompsonc S. thompsonc S. thompson S. thompsone S. thompsonc S. thompsonc S. thompsone S. thompsonc S. thompsonc S. thompson S.thompson S. thompson S. thompson S. thompson S. thompson S. thompson S. thompson S. thompson S. thompson S. blockley S. blockley S. blockley S. blockley S. blockley S. blockley S. blockley S. blockley 16 17 18 19 20 21 22 23 1 2 3 4 5 6 1 2 3 4 5 6 7 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 1 2 3 4 5 6 7 8 3/11/12/14 2/9/14 2 1/3/10/11/12/14/15 2/3/5/6/7/8/11/14 3/8 2/8/14 3/10/15 14 3/11 2/3/11 3/15 3/5/11/12/14 3/11/14 1/2/10 2/4/7/10/12/13/14 10/14 2 1/10 1/3/12/14 14 3/5/6/7/8/10/11/14/15 5/6/7/10/11/14 3/5/6/7/8/10/11/12/13/14 3/5/6/7/8/11/12/14/15 3/5/6/7/8/11/14 3/5/6/7/8/11/14 3/5/6/7/8/11/14 3/5/6/7/8/11/14 3/5/6/7/8/11/14 3/5/6/7/8/11/14 3/5/6/7/8/11/14 3/5/6/7/8/11/12/14 3/5/6/7/8/11/14 3/5/6/7/8/11/14 3/5/6/7/8/11/14 3/5/6/7/8/11/14 3/5/6/7/8/11/14 3/5/6/7/8/11/14 5/7/8/9 1/3/5/6/7/8/11/14 3/5/6/7/8/11/14/15 1/3/5/6/7/10/11/15 1/2/3/4/5/6/7/10/11/12/14/ 15 3/5/6/7/8/11 2/3/5/6/7/8/11 1/3/5/6/7/8/10/11/12/14 1/2/3/5/6/7/8/11/14 1/2/3/5/6/7/8/11/13/14 4/10/11/12/15 3/4/10/11/15 1/3/4/10/11/15 4/10/12/15 1/2/4/10/11/15 4/10/11/12/13/15 1/3/4/10/11/12/15 1/3/4/10/11/14/15

VOL. 3, 1976 NOTES 215

on February 7, 2020 by guest

http://jcm.asm.org/

TABLE 2-Continued

Salmonellagroup Cl Culture Patterns Salmonella group Cl Culture Patterns

type

strains

no. type

strains

no.

S. blockley 9 4/12 S. muenchen 1 1/10/12/13/14/15

S. blockley 10 1/11/13/14/15 S.muenchen 2 1/3/4/9/10/11/12/13/15

S. kentucky 1 3/10/11 S. newport 16 1/12/13/15

S.kentucky 2 9/10/11/13 S. newport 17 1/4/7/10/11/12/14/15

S.kentucky 3 10/11 S. newport 18 1/10/15

S. kentucky 4 11/14 S. newport 19 12/13/14

S. kentucky 5 3/11/12 S. newport 20 9/12/13/14/15

S. kentucky 6 3/9/11 S. newport 21 3/11

S. kentucky 7 10 S. newport 22 1/4/10/11/14/15

S. newport 23 14

S. kottbus 1 1/4/10/12/13/15 S. newport 24 1/4/10/12/13/14/15

S. kottbus 2 1/9/10/15 S. newport 25 4/9/10/12/13/14/15

S. kottbus 3 1/4/9/14/15 S. newport 26 1/4/10/15

S. kottbusb 4 4/10/12/13/15 S.newport 27 12/13/14/15

S. kottbusb 5 4/10/12/13/15 S. newport 28 1/3/4/10/11/13/14/15

S.kottbusb 6 4/10/12/13/15 S. newport 29 10/14/15

S. kottbusb 7 4/10/12/13/15 S. newport 30 1/4/11/12/13/15

S.kottbus' 8 4/10/12/13/15 S. newport 31 10/12/13/14/15

S. kottbusb 9 4/10/12/13/15 S. newport 32 12/13

S. kottbusr 10 4/10/12/13/15 S. newport 33 15

S. kottbusr 11 4/10/12/13/15 S.newport 34 2/10/14/15

S. kottbusr 12 4/10/12/13/15 S.newport 35 10/14 S. manhattan 1 1/4/10/11/12/15 S.newport 36 3/10/12/13/15

aOnly strongreactions (++ + orabove)arerecorded.

bCultures were isolated from a hospital outbreak.

r Cultureswereisolated fromacampusoutbreak.

detectable.

A

small

quantity

of the broth

cul-ture was

then

flooded

onto a nutrient agar

plate,

allowed

to

dry for

approximately

15min,

and

then

spotted with

phages using

a1-ml

sy-ringe

with

a 26-gauge

needle. The

plates

were

incubated overnight

at 37C

and read the

next

day. The cultures

were

examined with the

aid

of

an x10

aplanatic hand

lens

and viewed

through

the

bottom

of the

plate.

Phage activity

was

recorded

on

the

basis

of the reactions

de-scribed

in Table 2. The serotypes used in

this

study and the ensuing

patterns

are

listed

in

Table 2.

Itappears, from our initial results,

that

spe-cific phage

sets are not necessary to type

indi-vidual

serotypes.

Indeed,

acommon

well-devel-oped

set ortwomay be

adequate

tocharacterize

most

commonly encountered

salmonellae.

Over 1,600

Salmonella serotypes

have been

identified

to date. All share the

potential

for

sudden,

and

unexpected, prominence. Under

the

circumstances it is

inconceivable

that a

spe-cific

set

of

phages

will

always

be available

for

any given serotype.

Given the

universality

of

salmonellosis

and

the

realization

that a

large

variety of serotypes

are routinely isolated, the convenience and

availability of a

single collection of phages,

for

typing Salmonella in

general,

assumes a

sig-nificant dimension.

Appreciation isexpressed to Jacqueline Hunter for her mostvaluable laboratory assistance, and to Billie0.

Black-burn of the National Animal Disease Laboratory, Ames, Iowa.

LITERATURE CITED

1. Anderson, E. S. 1964. Phage typing of Salmonella other than S. typhi, p. 89-110. In Van Oye (ed.), The world problem of salmonellosis. Junk, The Hague.

2. Banker, D. D. 1955. Paratyphoid a phage typing.

Na-ture(London) 175:309-310.

3. Callow, B. R. 1959. A new phage-typing scheme for

Salmonellatyphimurium.J.Hyg. 57:346-359.

4. Craigie, J., and C. H. Yen. 1938. The demonstration of types ofB.typhosus by means of preparations of type II Vi phage. I. Principles and technique. Can. J. Public Health 29:448-484.

5. Craigie, J., and C. H. Yen. 1938. The demonstration of types ofB.typhosus by means of preparations of type IIVi phage. II. The stability and epidemiological significance ofVform types ofB.typhosus. Can. J. Public Health 29:484-496.

6. Gershman, M. 1972. Preliminary report: asystemfor

typing Salmonella thompson. Appl. Microbiol. 23:831-832.

7. Gershman,M. 1974. Aphage typing system for

Salmo-nella newport. Can.J.Microbiol.20:769-771. 8. Guinee, P. A. M., W. J. Van Leeuwen, and D. Pruys.

1974.Phage Typing of S. typhimuriuminthe Nether-lands. I.Thephage typing system. Zentralbl. Bakte-riol. Parasitenkd. Infektionskr. Hyg. Abt. 1 Orig.

ReiheA. 226:194-200.

9. Lilleengen, K. 1950. Typing of Salmonella dublin and Salmonella enteritidis by means ofbacteriophage.

Acta Pathol.Microbiol. Scand.27:625-640. 10. Lilleengen,D. 1952. Typing of Salmonellagallinarum

216 NOTES J. CLIN. MICROBIOL.

on February 7, 2020 by guest

http://jcm.asm.org/

NOTES 217

andSalmonella pullorum bymeansof bacteriophage.

Acta Path. Microbiol. Scand. 30:194-202.

11. Petrow, S., S. S. S. Kastiya, J. Pelietier, H. W.

Acker-mann, andJ. Peloquin. 1974. Phage typing scheme

forSalmonella newport. Ann. Microbiol. Inst. Pas-teur125:433-445.

12. Sechter, I., and C. B. Gerichter. 1968. Phage typing

schemefor Salmonella braenderup. Appl. Microbiol.

16:1708-1712.

13. Sechter, I., and C. B. Gerichter. 1969. Phage typing scheme for Salmonella blockley. Ann. Inst. Pasteur

Paris116:190-199.

14. Wilson, V. R., G. J. Herman, and A. Balows. 1971. Reportofanewsystemfor typingSalmonella

typhi-murium in the United States. Appl. Microbiol. 21:774-776.

VOL. 3, 1976

on February 7, 2020 by guest

http://jcm.asm.org/

ERRATA

Phage Typing

Set for

Group

C1

and

C2 Salmonellae

M. GERSHMAN

Department ofMicrobiology and Animal and Veterinary Sciences,UniversityofMaine, Orono, Maine04473

Volume 3, no. 2, p. 214,

column

2,

paragraph

1,line 8:

Change "enteritidis

(9), S. gallinarum,

and S.pullorum" to read

"enteritidis

(9),

S.

newport (11),

S. gallinarum,

and S.

pullorum

. . .

p. 214, column 2,

paragraph

2, line 3:

Delete

"11."

p. 215,

column

2:

Cultures beginning

with "S.

blockley"

should

appear

under the heading of

Salmonella group

Co

type strains.

p. 216, column 1: "S.

kottbus" cultures

no. 10, 11, and 12" should all read "S. kottbusb ...

Clinical Comparison

of

Aerobic,

Hypertonic, and Anaerobic

Culture

Media for the

Radiometric

Detection of

Bacteremia

R. MARIE COLEMAN, W. WAYNE LASLIE, AND D. W. LAMBE, JR.*

School of Allied Health andDepartment ofPathology andLaboratory Medicine,* Emory University,

Atlanta,

Georgia

30322

Volume

3, no. 3, p. 284, Table 5,

column

4:

Change

"Gram-positive rods"

to

"Gram-positive

cocci."