Journal of Applied Bacteriology 1995, 79, 360-367

Efficiency of different enrichment and isolation procedures for

the detection of Salmonella serotypes in edible offal
G. Arroyo and J.A. Arroyo'

'

Departamento de Microbiologia 11, Facultad de Farmacia, and Departamento de Microbiologia 111, Facultad de
Biologia Universidad Complutense Madrid, Spain
I

5209/01/95: received 24 January 1995, revised 5 April 1995 and accepted 10 April 1995
G . A R R O Y O A N D J . A . A R R O Y O . 1995. Rapid detection systems for Salmonella in foodstuffs are
currently being developed. However, existing standards still call for application of traditional
methods employing pre-enrichment followed by selective enrichment and isolation. T h e
efficacy of various methods was tested using 264 chicken and lamb organ meats.
Pre-enrichment was carried out in Tryptone Soy Broth (TSB) and enrichment in
Tetrathionate Brilliant Green Broth (TTB) at 37"C, Selenite Broth with Brilliant Green and
Sulphapyridine at 37C and 43"C, and Rappaport-Vassiliadis Broth (RV 10) at 42C. T h e
isolation media were Brilliant Green Agar (BGA), Deoxycholate Citrate Agar, Hektoen
Enteric Agar (HEA) and Salmonella-Shigella Agar.
Enrichment in RV/42"C followed by isolation on BGA as recommended by I S 0 standard
no. 6579 and enrichment in TTB/37"C followed by isolation in HEA, no longer
recommended by that standard, produced the best results. Low percentages of positive
samples and difficulties in detecting Salmonella are the result of interference by competing
organisms (Enterobacteriaceae) and the number of salmonellas present after enrichment.
A total of 528 samples (TSB, eggs, lamb liver and chicken liver) were inoculated with
Salm. enteritidis, Salm. kapemba and Salm. virchow, and the preceding experiment was
repeated. All the TSB and egg samples tested positive, but the percentage of positive samples
from the lamb and chicken liver was only 81-92%. Recovery of the salmonellas did not
depend upon the method employed or the serotype inoculated but instead on interference by
competing flora and the numbers of Salmonella present in the samples.

INTRODUCTION

Salmonellas are responsible for most cases of food poisoning in the developed world. Foods such as meat, eggs,
poultry and organ meats are common vehicles of salmonellosis. For that reason, rapid detection methods based primarily on immunological and genetic characteristics are
under development. However, the standards presently in
force in many countries recommend traditional methods,
which are slower, requiring a t least 5 d, though they can be
quite reliable when applied by experienced laboratory technicians.
A variety of methods are in use, and their success rates
depend upon a number of different factors. They employ
pre-enrichment in buffered peptone water, followed by
Correspondence to : Dr

G.Arroyo, Julian Romea 9, 28003-Madrid, Spain.

selective enrichment in more than one medium, usually

Muller-Kauffmann Tetrathionate Broth and Selenite
Cystine Broth incubated at 37C or at 43"C, and subsequently selective isolation on various solid media (Anon.
1981). Van Schotthorst et al. (1987) recommended enrichment in Rappaport-Vassiliadis (RV) broth a t 43"C, and
Beckers et al. (1987b) proposed replacing the Tetrathionate
method with the RV method in the IS0 standard (Anon.
1990).
The selective effects of the different media are mainly
based on the addition of substances that inhibit the growth
of contaminating microflora and prevent the proliferation of
competing microflora, on the incubation temperature and
the inoculum size employed. Studies on some of these
inhibitors (Arroyo and Arroyo 1995) have often yielded disappointing results. The numbers of competing Entcrobacteriaceae and Pseudomonadaceae in many natural foodstuffs

0 1995 The

Society for Applied Bacteriology

METHODS FOR SALMONELLA DETECTION 301

must be assumed to be higher than the numbers of Salmonella, and those competing organisms are also capable of
withstanding the same inhibitor concentrations as salmonellas during pre-enrichment and isolation, thus masking the
presence of salmonellas and giving rise to false positives
(Rhodes and Quesnel 1986).
The addition of stains, e.g. brilliant green, to the preenrichment media combined with raising the incubation
temperature to 43C has produced varying results for
Tetrathionate Broth (Van Schothorst et al. 1977). Arroyo
(1990) reported that temperature affected the growth of
certain Salmonella serotypes, making detection more difficult.
The use of antibiotics and chemical therapeutic agents in
the media has also been proposed. Osborne and Stokes
(1955) recommended adding sulphapyridine to enhance the
effectiveness of Selenite Broth.
The success of the isolation media depends basically
upon the enrichment step employed and the number of
competing organisms that survive that step.
Besides the international standards, the recommendations of the National Reference Center at the Carlos I11
Health Institute in Madrid are usually followed when analysing food and drink in Spain (Pascual Anderson 1993).
The object of the present study was therefore to evaluate
the efficacy of the recommended enrichment media and the
influence of incubation temperature using Selenite Broth
with added brilliant green and sulphapyridine. The efficacy
of four commonly used solid isolation media was also
tested.
The foodstuffs employed in this study were typical naturally contaminated sources of salmonellas, namely, lamb
organ meats and chicken liver, purchased at markets in
Madrid (Experiment 1). Tryptone Soy Broth (TSB), eggs
and lamb and chicken liver were also artificially contaminated with three Salmonella serotypes (Experiment 2).

MATERIALS AND METHODS

Experiment 1 : Naturally contaminatedsamples

A total of 264 organ meats purchased retail at 10 markets in

Madrid were examined. The organs used were chicken
livers (78) and lamb organs (186), including liver (46), lung
(46), heart (46), oesophagus (46), and spleen (2). The organ
meats were transported to the laboratory in a portable
cooler at 4C. Examinations were performed on the day of
purchase.
Twenty-five grams of each sample were weighed out
using sterile instruments under aseptic conditions, added to

0 1995 The Society for

225 ml of Tryptone Soy Broth (TSB), and homogenized in

sterile bags in a Stomacher model 400 Lab Blender for 2
min.

Microbiological examinations

The microbiological examinations employed differed somewhat from standard methods.

(a) Pre-enrichment was performed in T S B incubated at
37C for 18 h (Arroyo 1990).
(b) Selective enrichment was performed by transferring 10
ml of the pre-enrichment culture to a flask containing
100 ml of Muller-Kauffmann Tetrathionate Broth
(TTB) containing 1 ml of a 0.1% (w/v) solution of brilliant green, followed by incubation at 37C for 24 h.
Two flasks, each containing 100 ml of Selenite Broth
(SC) containing 1 ml of a 0.1 % (w/v) solution of brilliant
green and sulphapyridine were likewise inoculated with
10 ml of the pre-enrichment homogenate; one of the
flasks was incubated at 37"C, the other at 43"C, for 24 h
(Arroyo 1990). Finally, 0.1 ml of pre-enrichment
medium was transferred to 10 ml Rappaport-Vassiliadis
Broth (RV 10) (Vassiliadis 1983) and incubated at 42C
for 24 h.
(c) Selective isolation was performed by streaking the surfaces of plates containing Brilliant Green Agar (BGA),
Hektoen Enteric Agar (HEA), Salmonella-Shigella Agar
(SSA) and Deoxycholate Citrate Agar (DCA) with the
T T B and S C selective enrichment media. The plates
were all incubated at 37C for 24-48 h. T h e RV selective
enrichment medium was inoculated only onto BGA
(Vassiliadis et al. 1978) and incubated at 37C for 24-48
h. Colonies suspected of belonging to the genus Salmonella were isolated in test tubes containing Tryptone Soy
Agar (TSA) incubated at 37C for 24-48 h.
(d) Identification of salmonellas was carried out using morphological, biochemical (Holt 1993) and serological tests.
Strains identified as Salmonella were serotyped at the
National Reference Center at the Carlos I11 Health Institute in Madrid.

Experiment 2: Artificially contaminated samples

Pure cultures. Pure cultures of Salm. enteritidis, Salm.
kapemba and Salm. virchow isolated from lamb and chicken

organ meats were grown on TSA slants for 24 h and then

washed from the slant with sterile physiological saline solution. The suspension was diluted with sterile saline solution
to an optical density of 0.2 at 540 nm measured with a
Bausch and Lomb model Spectronic 20 spectrophotometer.
Serial dilutions were prepared from this stock suspension to

Applied Bacteriology, Journal of Applied Bacteriology 79, 360-367

362 G .

ARROYO AND J . A . ARROYO

yield the appropriate number of cells. That number was

determined by plating 0.1 ml of the diluted suspension on
TSA plates and incubating at 37C for 24 h.

Table 2 Efticiency of the four enrichment procedures in terms of

the number of isolated serotypes

I n o c u l a t i o n . Inoculation took place as follows :

TTB/37"C*

SC137"C

(a) Three sets of 44 samples of 225 ml of T S B were each

inoculated with cultures containing 1000 cells of S a l m .
enteritidis, Salm. kapemba and S a l m . virchow, respectively. These 132 samples were incubated at 37C for 18
h. Inoculation of the enrichment and isolation media and
detection of salmonellas were then as already described
in Experiment 1.
(b) A total of 132 pasteurized eggs purchased at 10 markets
in Madrid were washed in sterile water under aseptic
conditions, cracked and the contents of each egg homogenized in 225 ml of TSB. Three groups of 44 of these
samples were then inoculated with cultures containing
1000 cells of each of the three aforementioned serotypes,
respectively, and incubated at 37C for 18 h. Inoculation
of the enrichment and isolation media and detection of
salmonellas were then as already described in Experiment 1.
(c) A total of 132 samples of each lamb liver and chicken
liver, purchased at 10 markets in Madrid, were homogenized in a Stomacher model 400 Lab Blender (each
sample consisting of 25 g of organ meat in 225 ml of
TSB). Three groups of 44 samples of each of these two
organ meats were inoculated with cultures containing
1000 cells of each of the aforementioned serotypes,
respectively, and incubated at 37C for 18 h. Inoculation

Table 1 Number and percentage of the different Salmonella

serotypes detected in the 260 processed and confirmed strains

Serotype

No.

Salm. enteritidis
typhimurium
virchow
worthington
infantis
kapemba
give
brandenburg
havana
anatum
arizona
agona
cubana
Salm. autoagglutinable
14 serotypes

25
22
110
45
15
12
7
3
4
2
7
3
1
4
260

9.61
8.46
42.30
17.30
5.76
4.61
2.69
1.15
1.53
0.76
2.69
1.15
0.38
1.53
99.92

Enrichment

YO

Serotype

No.

47

18.07

Salm. enteritidis
typ himurium
virchow
worthington
kapemba
give
infantis
cubana
Salm. autoagglut.

10
7
14
4
3
3
3
1
2

96

36.92

enteritidis
typhimurium
virchow
worthington
kapemba
give
havana
arizona

4
7

Isolationt

40
25
5
4
4
7

SC/43"C

58

22.30

enteritidis
virchow
worthington
kapemba
Salm. autoagglut.

6
37
12
1
2

RV142"C

59

22.69

enteritidis
typhimurium
virchow
morthington
kapemba
infantis
brandenburg
anatum
agona

260

99.98

5
8
19
4
3
12
3
2
3
260

Total

* Enrichment broths and incubation temperature : TTB, Tetrathionate Brilliant Green Broth, 37C; SC, Selenite Broth (with
brilliant green and sulphapyridine), 37C and 43C; RV,
Rappaport-Vassiliadis Broth (RV lo), 42C.
t Number and percentage of positive isolation.
of the enrichment and isolation media and detection of
salmonellas were as already described in Experiment 1 .
RESULTS

Experiment 1

Of the 264 lamb and chicken organ meats examined, 83

(31.43%) tested positive for Salmonella. A total of 260
strains isolated were confirmed to be one of 14 serotypes of

0 1995 The Society for Applied Bacteriology, Journal of Applied Bacteriology 79, 360-367

METHODS FOR

Table 3 Efficiency of enrichment and

isolation procedures in terms of positive

strains of Salmonella detected

S A L M O N E L L A DETECTION 363

Isolation procedure7
Enrichment procedure

BGA

DCA

HEA

SSA

No. (%)

TTB/37"C*

7
(2.69)
14
(5.38)
13
(5)
59
(22.69)
93
(35.76)

15
(5.76)
27
(10.38)
15
(5.76)

15
(5.76)
28
(10.76)
12
(4.61)

10
(3.84)
27
(10.38)
18
(6.92)

57
(21.9)

55
(21.13)

55
(21.13)

47
(18.05)
96
(36.9)
58
(22.29)
59
(22.69)
260
(99.93)

SC/37"C
SC/43"C
RV/42"C
No.

("/.I

* Enrichment broths and incubation temperature : TTB, Tetrathionate Brilliant Green

Broth, 37C; SC, Selenite Broth (with brilliant green and sulphapyridine), 37C and 43C;
RV, Rappaport-Vassiliadis Broth (RV lo), 42C.
t Isolation media: BGA, Brilliant Green Agar; DCA, Deoxycholate Citrate Agar; HEA,
Hektoen Enteric Agar; SSA, Salmonella-Shigella Agar.
No., Number of strains detected.
Percentage of positive isolations given in parentheses.
Table 4 Serotypes detected according to the enrichment and isolation procedures used

Isolation procedure
Enrichment
procedure
TTB/37"C

SC/37"C

SC/43"C

RV/42"C

BGA

DCA

HEA

SSA

Salm. typhimurium
virchow

Salm . enteritidis
typhimurium
virchow
worthington
infantis
give
Salm. autoagglut.

Salm. enteritidis
typhimurium
virchow
worthington
infantis
give
cubana
Salm. autoagglut.
virchow
worthington
kapemba
typhimurium

Salm . typhimurium
virchow
kapemba
infantis
give

enteritidis
virchow
worthington
give
arizona

virchow
worthington
Salm. autoagglut.
enteritidis
typhimurium
virchow
worthington
kapemba
infantis
brandenburg
anatum
agona

virchow
worthington
arizona

enteritidis
virchow
worthington

virchow
worthington
kapemba

For abbreviations see Table 3.

0 1995 The Society for Applied

Bacteriology, Journal of Applied

Bacteriology

78, 360-367

enteritidis
virchow
worthington
kapemba
give
havana
arizona
enteritidis
virchow
worthington

364 G . ARROYO AND J . A . ARROYO

Salmonella. The distribution of serotypes, including

autoagglutinable strains, appears in Table 1.
Table 2 shows that 96 Salmonella strains (36.92%) were
detected using the SC enrichment medium incubated at
37C. Incubation of this same medium at 43C resulted in
only 58 Salmonella strains (22.30%). Enrichment in T T B
incubated at 37C yielded 47 positive strains (18.07%),
while enrichment in RV 10 incubated at 42C yielded 59
(22.69%) positive isolations. Table 2 also lists the different
serotypes detected using each of the media. TTB/37"C and
RV/42"C produced nine of the 14 serotypes detected and
SC/37"C produced eight. SC/43"C yielded the fewest
serotypes (5).
Salmonella virchow was the most abundant serotype and
was detected with all four of the enrichment methods used.
On the other hand, not all the minor serotypes present were
detected by all the different methods.
Table 3 gives the number and percentage of positive isolations on the solid media for each of the selective enrichment media employed. BGA yielded 93 positive isolations
(35.76%). Detection rates differed substantially, with TTB/
37C yielding only seven positive isolations (2.69%) and
RV/42"C yielding 59 (22.69%).T h e results obtained using
enrichment in SC/37"C and SC/43"C were quite similar, 14
(538%) and 13 (5%) positive isolations, respectively. The
Sample

Enrichment procedure
TTB/37OC sc-37"~

Inocula

Nature

No.

Salm. enteritidis

TSB

4 4 4 4
(100)
4 4 4 4
(loo)
44
38
(86.4)
44
36
(81.8)
4 4 4 4
(loo)
4 4 4 4
(100)
44
38
(86.4)
44
38
(86.4)
4 4 4 4
(100)
4 4 4 4
(100)
44
39
(88.6)
44
39
(88.6)

Eggs
Lamb liver
Chicken liver
Salm. kapemba

TSB
Eggs

Lamb liver
Chicken liver

Salm. virchow

TSB
Eggs

Lamb liver
Chicken liver

44
(100)
44
(100)
39
(88.6)
38
(86.4)
44
(100)

44
(100)

40
(90.9)
39
(88.6)
44
(loo)
44
(100)
40
(90.9)
41
(92.3)

results obtained using DCA, HEA and SSA were more

uniform. T h e best results, 27 (10.38%) and 28 (10.76%)
positive isolations, were achieved following enrichment in
SC/37"C; results were not as good when enrichment was
carried out in SC/43"C.
In terms of the variety of serotypes detected (Table 4),
the best results were obtained by BGA after enrichment in
RV/42"C (9 serotypes) and HEA after enrichment in TTB/
37C (8 serotypes). On the whole, interference by competing organisms on the isolation media was lower
following enrichment in T T B and RV.
Experlment 2

Table 5 presents the recovery of Salm. enteritidis, Salm.

kapemba and Salm. virchow in the different samples. The
results show that direct inoculation with 1000 cells in T S B
yielded a recovery rate of 100% positive samples, irrespective of the enrichment and isolation method used and the
serotype inoculated.
Inoculation of a foodstuff devoid or with negligible levels
of contamination, namely, pasteurized eggs, with 1000 cells
and homogenization with T S B also produced a 100%
recovery rate. As in the preceding case, the recovery rate
was the same for all the serotypes inoculated.

SC/~~OC
RV/42OC

44
(100)
44
(loo)
37
(84.1)
36
(81.8)
44
(100)
44
(100)
37
(84.1)
38
(86.4)
44
(loo)
44
(loo)
38
(86.4)
36
(81.8)

Table 5 Recovery of Salmonella

enteritidis, Salm. kapemba and Salm.
virchow using different procedures of
enrichment

44
(100)
44

(1W
40
(90.9)
37
(84.1)
44
(10)
44
(loo)
39
(88.6)
41
(92.3)
44

(W
44
41
(92.3)
39
(88.6)

Percentage of positive samples in parentheses.

0 1995 The Society for Applied Bacteriology, Journal of Applied Bacteriology 79, 360-367

METHODS FOR S A L M O N E L L A DETECTION 365

In contrast, inoculation of the lamb and chicken livers,

foodstuffs with normally high levels of contamination
(Enterobacteriaceae counts of up to lo7), with 1000 cells in
TSB homogenate yielded more uneven results, ranging
from a low of 81.81% positive isolations to a high of
92.27% positive isolations and a slight difference in favour of
RV/42"C. SC/43"C produced the lowest recovery rates.
Again, the results were independent of the salmonella
serotype inoculated.
The results obtained for the selective isolation media
tested in turn depended upon the enrichment broth
employed. On the whole, interference by competing
organisms was high, particularly on the BGA, making
detection of Salmonella difficult even though the samples
had been inoculated with that bacterium.
DISCUSSION

The success of enrichment and isolation media is based on

the presence of inhibitors intended to act against Grampositive contaminating microflora and Gram-negative competitive flora, normally Enterobacteriaceae. A study of 12
inhibitors a t similar concentrations (Arroyo and Arroyo
1995) demonstrated that the level of inhibition on salmonellas was the same as on the other Enterobacteriaceae examined and accordingly that inhibition of the multiplication of
such organisms during enrichment was difficult. Direct
counts of total Enterobacteriaceae in lamb and chicken
organ meats on Violet-Red-Bile-Glucose-Agar (VRBG)
reached up to lo7 cfu g - ' (Arroyo and Arroyo 1995). The
high initial levels of Enterobacteriaceae and their further
growth in the enrichment media masked the presence of
Salmonella, giving rise to false positives (Watson and
Walker 1978; Rhodes and Quesnel 1986). Salmonellas
cannot begin to multiply during enrichment until the
number of competitors has fallen (Van Schothorst and
Renaud 1983; Rhodes and Quesnel 1986). This decrease is
brought about by a fall in pH and depends upon the foodstuff in question, bacterial metabolism, and the presence of
large numbers of Gram-positive bacteria, including lactic
acid bacteria (Van Schothorst and Renaud 1985). In the
samples examined by the authors, there were rather high
numbers of Enterobacteriaceae with low numbers of Grampositive organisms, and the p H did not fall below 6.2
(unpublished data).
The results for the enrichment media in Experiment 1
varied. The greatest variety of different Salmonella
serotypes were detected using RV 10 (Vassiliadis et al.
1981; Vassiliadis 1983) and the largest number of positive
isolations using SC/37"C, though it should be noted that
most of these belonged to the two most abundant serotypes,
Salm. virchow and Salm. worthington, and that detection of
the minor serotypes using this latter medium was more dif-

ficult. Fagerberg and Avens (1976) reported that certain

serotypes were easier to detect in certain media than in
others.
TTB, which Beckers et al. (1987a, b) recommended
replacing with RV in the I S 0 standard (Anon. 1990), produced the fewest positive isolations, but the number of different serotypes detected was higher than using SC/37"C.
This suggests that certain inhibitors may affect the growth
and multiplication of some of the minor serotypes present.
However, according to Van Schothorst et al. (1977), inhibition depends less upon the serotype than upon the sensitivity of certain cells to the inhibitor. Patil and Parhad
(1986) reported that T T B was the best enrichment medium
when E. coli and other lactose-positive Enterobacteriaceae
were the predominant competing organisms, whereas SC
yielded better results when organisms of the genus Proteus
predominated.
Raising the incubation temperature may increase the
number of positive isolations (Carlson and Snoeyenbos
1972). However, tetrathionate with brilliant green at 43C
is toxic to many salmonellas (Harvey and Price 1979;
Arroyo 1990). Both the number of positive isolations and
the variety of serotypes detected decreased using SC/43"C.
Studies with pure cultures have indicated that certain Salmonella serotypes cannot multiply at high incubation temperatures (Carlson and Snoeyenbos 1974; Van Schothorst
et al. 1977). On the other hand, survival of most of the
Enterobacteriaceae at 43C and at 44C (e.g. E. coli, whose
presence as a faecal coliform is detected in lactose broth
incubated at 44"C), made isolation of salmonellas extremely
difficult. Van Schothorst and Renaud (1983) reported that
isolation of salmonellas was virtually impossible when the
number of lactose-positive Enterobacteriaceae was 10' g or ml-', hence raising the temperature would appear to be
unnecessary when using the SC medium.
Isolation on BGA following enrichment in SC/37"C and
SC/43"C was quite difficult because of the large numbers
of competing bacteria present. For the salmonellas to be
detectable on that medium, their concentration in the
enrichment medium after incubation must reach a level of
lo3 ml- (Van Leusden et al. 1982); at lower levels they
remain undetectable, even if the level of the competing
flora drops to below lo3 ml - '.
Detection on BGA is facilitated by greater inhibition of
competing organisms in RV 10 (Vassiliadis 1983). Competing bacteria belong mainly to the genera Proteus, Morganella and Shigella. The presence of Yersinia is important,
because the Rappaport-Wauters medium is used for enrichment of that genus. That medium is similar to RV, and the
colonies of that genus on BGA do not differ in appearance
from Salmonella colonies.
Isolation on HEA and DCA after enrichment in T T B
yielded the most positive isolations, and anaerobic culture is

'

'

R ? 1995 The Society for Applied Bacteriology. Journal of Applied Bacteriology 79, 360-367

366 G. A R R O Y O A N D J . A . A R R O Y O

not required when using DCA (Edgar and Soar 1979).

However, contradicting the findings of Moriiiigo et al.
(1989), these media did not inhibit the growth of Pseudomonas aeruginosa or other Pseudomonadaceae and Vibrionaceae. Based on these results, RV would appear to be the
most appropriate enrichment medium, but complete rejection of T T B would appear to be premature, since that
medium yielded the greatest variety in the number of
serotypes and lower interference by competing organisms
than SC.
The results of Experiment 2 corroborated the results of
Experiment 1. In the T S B and commercial pasteurized egg
samples, in which there was no interference by competing
organisms, 100% of the samples tested positive, irrespective
of the serotype inoculated and the method employed, even
when lower concentrations of inocula were used (results not
presented here). Previously, inoculation with small
numbers of Salm. montevideo and S a l m . heidelberg yielded
excellent recoveries using Selenite Broth, whereas the concentration of the inoculum had to be increased to 500 cells
when using Tetrathionate Broth (Bailey et al. 1981). In the
present experiment the pre-enrichment broth was inoculated to facilitate the recovery of salmonellas in the T T B
(Taylor and Silliker 1962).
The level of competing organisms was high in the lamb
and chicken liver samples, and the percentage of positive
isolations was lower, though the results did not vary much
with the method used and the serotype inoculated. Accordingly, successful detection of salmonellas in those organ
meats was dependent upon interference by competitors.
The similar behaviour exhibited by Salm. kapemba, a minor
serotype in Experiment 1, indicates that it is able to tolerate
the same inhibitor concentrations as S a l m . enteritidis and
Salm. virchow and provides further confirmation that salmonella concentrations must be lo3 ml- after enrichment.
Difficulties in detecting salmonellas are heightened when
there are cross-reactions with other Enterobacteriaceae,
such as Proteus, Morganella and Citrobacter freundii.
Increasing interest in developing fast methods of analysis
and future application of those methods may help to mitigate these difficulties.
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Anon. (1990) International Organization for Standardization
Microbiology-General guidance on methods for the detection
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Arroyo, G. (1990) Estudio de las estirpes de Salmonella presentes
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