Phage Typing Set for E1, E2, E3, and E4 Salmonellae

(1)

Phage Typing

Set for E1, E2, E3,

and

E4

Salmonellae

M. GERSHMAN

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

Received for publication 7May 1976

EighteencommonserotypesrepresentativeofgroupE,, E2, E3, and E4

Salmo-nella werecharacterized using asinglesetof phages.

Salmonellae

are

commonly

implicated

in

casesof

food

poisoning

and

constitute one

of the

major problems confronting

health authorities

everywhere.

An

estimated

two to

four

million

people

are

infected

annually

in

the

United

States

alone

(5).

Outbreaks of salmonellosis,

however,

can

be

controlled, if

not

prevented,

if

the presence of the organism can be

established.

Many

outbreaks of this

disease have been

re-solved by

locating

the source

of

an

infection

by

serologically identifying the species involved

(2). However, if a

particular

species is

fairly

common

and

widely

distributed,

it

becomes

dif-ficult,

if not

impossible,

toassess itsrole in an

outbreak.

One of the

most

important

properties

of

bac-teriophage

is its

host

specificity

and

its

accom-panying

ability

to

distinguish

types among

ap-parently

identical

serotypes.

Epidemiological

use

has been

made

of this

property,

and

a

num-ber of

commonly

occurring

Salmonella

sero-types are now

routinely

characterized

through-out

the

world

by phage

typing (1).

In

the

course

of

our activities to

develop

phage typing

sets

for

some

of the

more

impor-tant

salmonellae,

itwas

observed that

one

sin-gle

set

of

phages

could be used

to

delineate

a

variety of

Salmonella

C,

and

C,

serotypes (4).

Given

this

revelation,

we

proceeded

to

develop

a

single

set

of

phages for typing Salmonella

serotypes

representative of

serological

groups

E1, E2, E3, and E4.

Using

a

collection of

Salmonella

anatum,

S.

binza, and S.

senftenberg

phages

developed

in

our

laboratory

(3;

Gershman,

J.

Milk

Food

Technol.,

inpress), we were

able

totype 412 of

the 420

Salmonella

El,

E2, E3, and

E4 cultures

examined

at

random. The

lytic

pattern of

these

phages

is

noted

in

Table

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

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 by using

a 1-ml

syringe with

a 26-gauge

needle. All of the

phages employed

in

this

study

were

used

at a

routinetest dilution of not less than 10-3. The

plates

were

incubated overnight

at

37°C

and

read the

The cultures were

examined

with

the

aid

of an X10

aplantic

hand lens and

viewed through the bottom of

the

plate. Phage

activity was

recorded

on

the

basis of the

reac-tions

described

in

Table

1.

The

serotypes

used

in

this study

and the

ensuing patterns are

listed

in

Table

2.

The

convenience

inherent

in

this

expanded

typing

set appears to

be

epidemiologically

sig-nificant.

Appreciation is expressed to Jacqueline Hunter for her mostvaluable laboratory assistance,and to Billie0. Black-burn of the National Animal Disease Laboratory, Ames, Iowa, and L. Le Minor of the Pasteur Institute, Paris, France, for their assistance in securing cultures for this project.

LITERATURE CITED

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

2. Gayler,G. E., R. A. MacCready, J. P. Reardon, and B. F. McKernan. 1955. An outbreak ofSalmonellosis tracedtowatermelon. Public Health Rep. 70:311-313.

3. Gershman,M. 1974. Aphage typing system for Salmo-nella anatum. Avian Dis. 18:565-568.

4. Gershman,M. 1976.Phage typing set for groupC,and C2Salmonellae. J. Clin. Microbiol.3:214-217.

5. Okey,C. H. 1967. Salmonellosis. J. Maine Med.Assoc.

58:85-94.

196

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J. CLIN. MICROBIOL.

TABLE 2. Reactions of test strains at routine testdilutionsa

Type strain Culture no. Pattern

Salmonella group EI S. amsterdam S.amsterdam S. amsterdam S. amsterdam S. amsterdam S. amsterdam S.amsterdam S. amsterdam S. amsterdam S. amsterdam S. amsterdam S. amsterdam

S.anatum S.anatum S.anatum S. anatum S.anatum S.anatum S.anatum S.anatum S.anatum S.anatum S.anatum S.anatum S.anatum S.anatum S.anatum

S. give S. give S.give S.give S. give S. give S. give S.give S. give

S. lexington

S. meleagridis

S. muenster S. muenster S. muenster S. muenster S. muenster S. muenster S. muenster S. muenster

S. uganda

S. westhampton

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

6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

1 2 3 4 5 6 7 8 9

1 1 1 2 3 4 5 6 7 8

1 1

1/6/7/13/16 6/7/13/16 1/6/7/16 1/6/7/13/17/18 1/6/7/13 1/7/13

1/6/7/13/16/17/18 1/2/3/6/7/16/18 6/7/13 7/13 1/6/7/10/16 7/13/16/17

1/2/3/4/6/7/8/9/10/12/15/16/18 1

1/4 2/4/6 6/17/18

1/2/4/5/6/7/8/9/10/12/13/15/16/18 1/2/4/5

1/2/4/5/6/7/9/10/13/16/18 7/13

1/2/3/4/5/6/7/8/9/10/12/13/15/16/18 1/2/4/5/6/9/10/13/16/17/18 1/2/3/4/6/7/9/10/11/13/16/18 1/2/3/4/6/7/8/9/10/12/13/15/16/18 1/2/3/4/5/6/7/9/10/13/14/16/18 1/2/3/4/5/6/7/8/9/10/12/13/14/15/16/18

1/6/7/13 1/4/6/7/9/10/16

1/2/4/5/6/7/9/10/12/15/16/17/18 6/7/13

6/7/13/17/18

1/2/4/6/7/9/10/12/15/16/17/18 1/2/3/4/5/6/7/9/12/13/14/16/17/18 6/7

2/6/7/13

6

1/4/7/9/10/13/16

1/2/4/5/6/7/9/10/16 1/2/4/5/6/7/9/10/13/16 1/2/3/4/6/7/9/10/13/16/18 2/3/5/9/10/13/16 1/2/3/4/5/7/9/10/13/16 1/2/4/5/7/9/10/13/16 2/3/4/5/7/9/10/13/16

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

1/2/4/5/6/7/8/9/10/12/15/16/17/18

16/18

198 NOTES

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Type strain Salmonella groupE2

S. binza S. binza S. binza S. binza S. binza S. binza S. binza S. binza S. binza S. binza S. binza S. binza S. binza S. binza S. binza S. binza S.drypool S.drypool S. drypool S.

drypool

S.drypool S.

drypool

S.drypool S.drypool S. drypool S.drypool S. halmstad S. halmstad S. manila S. newington S. newington S. newington S. newington S. newington S. newington S. newington S. newington S. newington S. newington

Salmonella groupE3 S. illinois S. illinois S. illinois S. illinois S. illinois S. illinois S.illinois S.minneapolis S.minneapolis S.minneapolis S.minneapolis Culture no. 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 1 2 3 4 5 6 7 8 9 10 1 2 1 1 2 3 4 5 6 7 8 9 10 1 2 3 4 5 6 7 1 2 3 4 Pattern 1/4/6/8/12 1/4/6/8/9/10 1/4/5/8/11/12 1/4/6/7/9/10/11 1/4/6/8/11 1/4/6/8 1/4/6/8/9/11 1/2/3/4/9/12 1/4/6/10/11 1/4/6/8/9/10/11 1/4/5/6/8/9/10/11/12/15/16 1/2/3/4/6/7/8/9/10/11/12/13/14/15/16 1/4/6/8/9/10/11/12/15/16 1/4/5/6/8/9/10/12/14/15/16 1/4/6/9/10/11/15/16 11 6/7/16 1/6/7/16 1/6/7/16/17 7 1/6/7 6/7/17 7/17 1/6/7/10/16 2/3/5/6/7 1/7/16 1/18 1/12/15 6 1/4/6/7/8/9/10/16 4/7 1/6/7/10/18 1/4/6/7/9/10/16/18 1/4/6/7/8/9/10/16/17 1/4/5/6/7/9/10/16/17/18 1/4/6/9/10/16/18 1/4/5/6/7/9/10/14/16 1/6/7/17 1/4/6/9/10/16 1/6 1/6/16 1/4/6/7/9/10/16 1/6/7/10/16 6 1/2/3/4/5/6/7/8/9/10/14/15/16 1/4/6/9/10/16 1/4/6/7/8/9/10/12/15/16/18 6/7 1/2/3/4/5/6/7/9/10/14/15/16/17/18 1/4/5/6/7/8/9/10/14/15/16/18

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J. CLIN.

TABLE 2-Continued

Typestrain Cultureno. Pattern S. thomasville 1 1/2/3/4/5/6/8/9/10/12/15/16

S.thomasville 2 1/2/4/5/6/7/8/9/10/11/12/13/14/15/16/18

S. thomasville 3 6

S. thomasville 4 6/7/11/16/17

S. thomasville 5 1/4/6/11/16

S. thomasville 6 1/4/5/6/8/9/10/11/14/15/16

S.thomasville 7 1/4/5/9/10/11/14/15/16

S. thomasville 8 1/6/11/16

S.thomasville 9 1/4/6/7/9/10/11/15/16/17/18

S.thomasville 10 1/6/11/14/15/16/17/18

Salmonella group E4

S.senftenberg 7 1/6/17/18

S.senftenberg 8 1/6/13/16/17

S.senftenberg 9 1/6/13/15/16/17/18

S.senftenberg 10 1/6/16

S. senftenberg 11 1/6/13/16

S.senftenberg 12 1/13

S.senftenberg 13 1/6/13/15/17

S.senftenberg 14 6

S.senftenberg 15 1/6/8/13/15/16/17

S. taksony 1 1/4/6/9/10

S.taksony 2 1/4/9/10/16

S. taksony 3 1/13

Only strong reactions (++ +orabove) arerecorded.

200 NOTES