International Journal of Food Microbiology 81 (2003) 169 173

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Short communication

Occurrence of salmonellae in retail chicken carcasses and their

products in Spain
lvarez-Astorga, Carlos Alonso-Calleja, Benito Moreno,
Rosa Capita*, Maite A
Mara del Camino Garca-Fernandez
Department of Food Hygiene and Food Technology, Veterinary Faculty, University of Leon,
Campus Universitario de Vegazana, s/n., 24071 Leon, Spain
Received 6 October 2001; received in revised form 8 March 2002; accepted 28 April 2002

Abstract
This study examined the incidence of Salmonella in Spanish poultry products. Samples included chicken carcasses, chicken
parts (wings, legs and gibletslivers and hearts) and processed chicken products (red sausages, white sausages and
hamburgers). The average detection rate was 49%, with the highest (55%) in chicken carcasses (skin) and the lowest (20%) in
hamburgers. The chicken carcasses purchased in supermarkets were more contaminated (75%) than those from poulterers shops
(25%). Salmonella Enteritidis, S. Poona, S. Paratyphi B and S. Worthington were isolated in 34.3%, 11.4%, 2.8% and 1.4% of
the samples, respectively. One (1.4%) red sausage sample harboured two serotypes (S. Enteritidis and S. Worthington). This fact
emphasizes the usefulness of subtyping several Salmonella isolates from the same sample in epidemiological studies.
D 2003 Elsevier Science B.V. All rights reserved.
Keywords: Salmonella; Incidence; Serotypes; Poultry meat; Chicken products; Spain

1. Introduction
Salmonellosis is the most prevalent foodborne
disease in many countries worldwide (DAoust,
1997). Chicken products are widely acknowledged
to be a significant reservoir for Salmonella and have
frequently been incriminated as a source of salmonel-

*
Corresponding author. Instituto de Ciencia y Tecnologa de
los Alimentos, C/La Serna, no. 56, 24007 Leon, Spain. Tel.: +34987-243123/238162; fax: +34-987-243123.
E-mail address: dhtrcg@unileon.es (R. Capita).

lae contamination (Mulder, 1999; Baeumler et al.,

2000). The incidence of Salmonella in poultry has
been well determined in many countries. However, in
Spain, no specific studies to determine the contamination rates of Salmonella in chicken products have
been published.
The aims of this study were (1) to determine the
prevalence of Salmonella in chicken products in
Spain, (2) to compare the level of contamination of
two groups of chicken carcasses (from supermarkets
and from poulterers shops) and (3) to survey the
Salmonella serotypes present in chicken meat and
their products.

0168-1605/03/$ - see front matter D 2003 Elsevier Science B.V. All rights reserved.
PII: S 0 1 6 8 - 1 6 0 5 ( 0 2 ) 0 0 1 9 5 - 2

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R. Capita et al. / International Journal of Food Microbiology 81 (2003) 169173

2. Materials and methods

2.1. Samples
A total of 40 chicken carcasses, 15 chicken parts
(five wings, five legs and five gibletslivers and
hearts) and 15 processed chicken products (five red
sausages, five white sausages and five hamburgers)
were collected at different locations in the northwest
Spain. Chicken carcasses were purchased from supermarkets (24) and from poulterers shops (16). Chicken
parts and also processed chicken products were purchased from poulterers shops. Each sample was
placed in a separate sterile plastic bag. Samples were
transported to the laboratory immediately after collection in a ice chest and tested upon arrival or stored at
2 jC for no longer than 4 h.
2.2. Microbiological analysis
Sampling, isolation and identification of strains
were made as previously reported (Capita et al.,
2001). A 25-g sample of each product was taken
aseptically by scalpel excision and placed in a stomacher bag containing 225 ml of nutrient broth (NB,
CM1, Oxoid, Hampshire, UK). For products containing skin (carcasses, wings and legs), 25 g of skin was
cut as a sample. For whole birds, the 25 g was made
up of samples of breast, wing, leg, neck, dorsal and
ventral skin. The samples and the NB were treated in a
Stomacher 400 (AJ Seward, London, UK) for 2 min
and incubated together in the bag for 24 h at 35 jC.
After incubation, 0.1 ml of the preenriched sample
was diluted in 10 ml of tetrathionate brillant green
(QA Laboratories, San Diego, CA, USA) and incubated at 35 jC in a waterbath. After 6 8 h, decimal
dilutions were prepared in peptone water (CM9,
Oxoid) and then 1 ml of the second dilution (10 2)
filtered through an Iso-GridR hydrophobic grid membrane filter (HGMF) with the aid of a sterile IsoGridR Filtration Unit and a vacuum pump, as developed by QA Laboratories (Anonymous, 1989). Filters
were aseptically transferred to the surface of predried
EF-18 medium plates (QA Laboratories) and incubated for 24 h at 42 jC. Four green, blue-green or blue
colonies (presumptive salmonellae) were selected in
each sample for subsequent identification on the basis
of the following tests: Gram stain (Harrigan and

McCance, 1976), catalase reaction, oxidase reaction,

oxidation/fermentation of glucose, motility test,
nitrate reduction (Cowan, 1974), sulphide production,
indole production, phenylalanine deaminase (Koneman et al., 1989), triple sugar iron (TSI), lysine iron
agar (LIA), citrate, methyl red, Voges Proskauer,
urease, ornitine and lysine decarboxylase, arginine
hydrolase (Ewing, 1986) and h-galactosidase (Anonymous, 1998a). The 280 strains were grouped in
biochemical profiles depending on the results in the
previous tests. The allocation to one particular genus
and species was initially done following the identification schemes proposed by Brenner (1984) and
Ewing (1986). Moreover, in order to confirm the
identifications carried out, one strain from each profile
was inoculated into microtubes of API 20E strips
(20,100, bioMerieux, Marcy-lEtoile, France) following the manufacturers instructions. The bacteria were
identified using the database provided by the manufacturer (API LAB Plus ver 3.2.2., bioMerieux).
Strains were serotyped in the Centro Nacional de
Microbiologa, Virologa e Inmunologa Sanitarias del
Instituto de Salud Carlos III (Majadahonda, Madrid,
Spain).

3. Results and discussion

Salmonellae were detected in 49% of the samples
(Table 1). The incidence of Salmonella in chicken
products obtained by other authors varied between 0
and 100% (Cox and Bayley, 1987; Bryan and Doyle,
1995; Waldroup, 1996). It should be mentioned that
isolation rates depend upon the country where the
study was carried out, the sampling plan and the
detection limit of the methodology (Roberts, 1982;
Uyttendaele et al., 1998). It must be indicated that the
HGMF method used in this study in theory allowed us
to detect one Salmonella cell per gram of sample
(Entis, 1981, 1986; Sharpe, 1994).
Contamination rates of chicken carcasses (55%)
were higher than those of chicken parts (40%). This
fact can be explained as breast skin has a high contamination rate, due to fecal contamination as broilers
lie on their breasts on the floor of the broiler house
(Shackelford, 1988). In addition to breast areas, whole
carcasses would retain heavily contaminated regions
such as the neck skin (Uyttendaele et al., 1998).

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171

Table 1
Rates of Salmonella contamination and frequency of serotypes in each class of chicken samples
Type of chicken sample

Salmonella

S. Enteritidis

S. Poona

S. Paratyphi B

S. Worthington

Chicken carcasses
From supermarkets
From poulteres shops
Chicken parts
Wings
Legs
Giblets
Processed chicken products
Red sausages
White sausages
Hamburgers
Total

22/40 (55%)a
18/24 (75%)
4/16 (25%)
6/15 (40%)
2/5 (40%)
2/5 (40%)
2/5 (40%)
6/15 (40%)
2/5 (40%)
3/5 (60%)
1/5 (20%)
34/70 (49%)

14
12
2
4
2
2

6
2
3
1
24/70 (34.3%)

8
6
2

8/70 (11.4%)

2/70 (2.8%)

1
1

1/70 (1.4%)

Number of Salmonella-positive samples/number of samples (percentage of Salmonella-positive samples).

Plummer et al. (1995) also found, as we did, higher

contamination rates in whole birds than in chicken parts
(wings and legs). On the other hand, as mentioned
above, Uyttendaele et al. (1998) show that the number
of Salmonella positive chicken parts exceeded those of
chicken carcasses. These authors have suggested the
importance of cross-contamination as the origin of
Salmonella contamination in poultry parts.
The incidence of Salmonella in processed chicken
products (40%) was the same as that found in chicken
parts and less than that in chicken carcasses. Even
though the production of sausages and hamburgers
requires a significant amount of handling of meat and
thus increases the risk of cross contamination, these
products contain spices and herbs that are antimicrobial
and could help to reduce contamination (Jay, 1992).
Contamination of raw meat with Salmonella is not
generally considered a risk to the consumer because
the food is expected to be heated sufficiently before
consumption thus eliminating the pathogen. Although
in the majority of countries this microorganism must
be absent from ready-to-eat food products, there are
few Microbiological Norms referring to the contamination of raw poultry products, even though they can
cause disease, either directly or indirectly by crosscontamination. There are no Standards in Spain referring to the contamination of refrigerated chicken
carcasses or chicken parts. Pascual Anderson (1982,
1992) establishes guidelines for microbiological contamination in poultry carcasses: Salmonella should be
absent in 25 g of sample. In some American states

standards are less strict than in Spain: absence in 1 g

sample in Iowa and Nebraska (Wehr, 1982). In France
the Centre Nationale dEtudes et de Recommendations sur la Nutrition et lAlimentation (CNERNACNRS, 1996) specifies for these food products
absence in 1 g of neck skin, with n = 5 and c = 2.
With respect to ground meat products, the Spanish
Microbiological Standards (Anonymous, 1998b)
specifies absence in 10 g of sample (with n = 5 and
c = 2).
As shown in Table 1, the contamination rate in
supermarket carcasses (75%) was higher than in
poulterers shops carcasses (25%), possibly due to
the short period of time that the chicken carcasses are
kept in the poulterers shops (generally less than 16 h)
because these establishments are supplied daily by
chicken slaughterhouses. These results differ from
those obtained by Plummer et al. (1995), who detected a lower number of Salmonella contaminated
carcasses from supermarkets (18.6%), than from poulterers shops (24.5%). These authors suggest that a
possible cause of this lower number is the greater use
of use by dates by the supermarkets and also of
packaging that would prevent further cross contamination between samples. Moreover, according to these
authors, there is also likely to be a fast turnover rate
in supermarket chicken.
Four different serotypes, S. Enteritidis, S. Poona, S.
Paratyphi B and S. Worthington, were found as seen
from Table 1. One sample of red sausages harboured
more than one Salmonella serotype (i.e. S. Enteritidis

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R. Capita et al. / International Journal of Food Microbiology 81 (2003) 169173

and S. Worthington). The presence of both serotypes in

the same red sausage sample emphasizes the usefulness
of subtyping several Salmonella isolates from the same
food sample in epidemiological studies.
The high percentage of samples with S. Enteritidis
coincides with the results obtained by other investigators (Abu-Ruwaida et al., 1994; Jerngklinchan et al.,
1994; Plummer et al., 1995; Telo et al., 1998; Uyttendaele et al., 1998; Zewde, 1999). The elevated presence
of this serotype in chicken products (including eggs)
agrees with S. Enteritidis being the predominant serotype associated with outbreaks due to the consumption
of eggs (Desenclos et al., 1996; Telo et al., 1998). In
Spain, this serotype is the most frequently isolated one
from poultry meat and it is the most prevalent found in
human cases of salmonellosis over the last few years
(Capita et al., 2000). The increase of S. Enteritidis in
poultry meat which has occurred over the last few years
and the consequent increase in human salmonellosis
cases caused by this serotype are due to the increase in
the incidence of S. Enteritidis in worldwide poultry
exploitations (Suzuki, 1994).
S. Poona was isolated from 11% of the chicken
samples examined and was detected only in chicken
carcasses. This serotype has been isolated from meat
and chicken products (Nabbut and Al-Nakhli, 1982;
Adesiyun et al., 1988; Jerngklinchan et al., 1994).
Nevertheless, S. Poona is more associated with water
than with food (DAoust et al., 1990; Shane et al.,
1990; Anonymous, 1991; Usera et al., 1991, 1995;
Golden et al., 1993).
S. Paratyphi B was isolated from two giblet
samples. Although it is not common to find this
serotype in chicken products, Jerngklinchan et al.
(1994) also isolated it from raw chicken meat and
liver samples in Thailand, and Zewde (1999) from
chicken carcasses and chicken pieces in Ethiopia.
S. Worthington was detected in a red sausage sample. None of the authors consulted isolated this serotype from chicken meat. Its presence in sausages could
be due to other different to chicken meat ingredients.
The results of this study indicate that poultry products could be considered a major potential source of
human salmonellosis, given the high incidence of S.
Enteritidis. For this reason, efforts should be made for
consumers to be informed and follow the basic instructions regarding storage temperature, cooking and prevention of contamination and cross-contamination.

Acknowledgements
This work has been financially supported by the
Spanish Comision Interministerial de Ciencia y
Tecnologa (CICYT) (reference no. ALI91-0294).
Dr. Capita was beneficiary of a fellowship from the
Spanish Ministerio de Educacion y Ciencia (PFPI).
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