This article appeared in a journal published by Elsevier.

The attached
copy is furnished to the author for internal non-commercial research
and education use, including for instruction at the authors institution
and sharing with colleagues.
Other uses, including reproduction and distribution, or selling or
licensing copies, or posting to personal, institutional or third party
websites are prohibited.
In most cases authors are permitted to post their version of the
article (e.g. in Word or Tex form) to their personal website or
institutional repository. Authors requiring further information
regarding Elsevier’s archiving and manuscript policies are
encouraged to visit:
http://www.elsevier.com/authorsrights
 Author's personal copy

Food Control 36 (2014) 199e204

Contents lists available at ScienceDirect

Food Control
journal homepage: www.elsevier.com/locate/foodcont

Antimicrobial susceptibility of Salmonella strains isolated from retail

meat products in Poland between 2008 and 2012
_
qukasz Ma˛ ka a, *, Elzbieta Ma  zy
ckiw a, Halina Scie _ n
 ska a, Kamila Paw1owska a,
Magdalena Popowska b
a
Laboratory of Food Microbiology, Department of Food Safety, National Institute of Public Health e National Institute of Hygiene, Chocimska 24,
00-791 Warsaw, Poland
b
Department of Applied Microbiology, Institute of Microbiology, University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland

a r t i c l e i n f o a b s t r a c t

Article history: A total of 106 Salmonella strains were isolated in the years 2008e2012 from retail meat products
Received 5 March 2013 sampled in Poland. Strains from poultry meat (n ¼ 81), pork (n ¼ 7), beef (n ¼ 3) and mixed meat
Received in revised form (n ¼ 15) were serotyped and subjected to antimicrobial susceptibility testing by the disk diffusion
13 August 2013
method (19 antibiotics). Twenty-one Salmonella serotypes were identified, with the three most
Accepted 20 August 2013
common being Salmonella Enteritidis (34.9%), Salmonella Infantis (14.2%) and Salmonella Typhimurium
(10.4%).
Keywords:
The majority of the Salmonella strains (68.9%; n ¼ 73) were resistant to one or more antimicrobial
Salmonella
Retail food
compound. Among the resistant isolates, 31 were resistant to one antibiotic, 4 to two, 10 to three, 13 to
Meat products four, and 15 to five or more antibiotics.
Prevalence Of the Salmonella Enteritidis isolates, 54% were resistant to at least one antibiotic, while much higher
Antimicrobial susceptibility frequencies of resistance were found in Salmonella Newport (100%), Salmonella Typhimurium (91%),
Salmonella Hadar (85.7%), Salmonella Virchow (80%) and Salmonella Infantis (80%).
The most common resistance observed among the Salmonella isolates was to nalidixic acid (52.8%). The
isolates were also frequently resistant to tetracycline (32.1%), ampicillin (28.3%), streptomycin (28.3%)
and sulphonamides (26.4%). All of the tested strains were susceptible to cefepime, cefotaxime, ceftazi-
dime, ceftriaxone, ciprofloxacin, ertapenem and imipenem.
Salmonella strains isolated from poultry meat showed the widest spectrum of resistance (to 12 of the
19 tested antimicrobials) compared with isolates from the other meat sources.
The level of resistance among Salmonella strains isolated between 2008 and 2012 was consistently
high: 59.1% in 2010, 84.6% in 2011 and 64.7% in 2012. In addition, there was an increase in the number of
multiresistant strains over this period, from 23.1% in 2010 to 81.8% in 2012. The demonstration that meat
products are a source of antibiotic resistant Salmonella strains is a serious concern for public health and
food safety.
Ó 2013 Elsevier Ltd. All rights reserved.

1. Introduction
Members of the bacterial genus Salmonella, classified within
the family Enterobacteriaceae, are among the most common food-
borne pathogens. Salmonellae are widespread in nature, being
present in domestic and wild animals as pathogens or
commensal microorganisms. These robust bacteria can survive in
the environment outside their hosts for long periods. More than
* Corresponding author. Tel.: þ48 225421384; fax: þ48 225421225.
E-mail address: lmaka@pzh.gov.pl (q. Ma˛ ka).
2500 serovars comprise the Salmonella genus (Grimont & Weill,
2007) and new serovars are regularly described
(Guibourdenche et al., 2010).
S. Enteritidis and S. Typhimurium are the most commonly re-
ported serovars in the European Union (EU), being associated with
52.3% and 23.3% of all confirmed human infections (salmonellosis),
respectively. Since 2006, S. Infantis has been the third most common
serovar in the EU (EFSA and ECDC, 2011). Over the same time period,
S. Enteritidis and S. Typhimurium represented over 80% of the Sal-
monella isolated from human cases in Poland. The percentage of cases
associated with S. Enteritidis decreased from 77.6% in 2006 to 68.5% in
2010. Moreover, in 2010, the second most prevalent serotype was

0956-7135/$ e see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodcont.2013.08.025
 Author's personal copy
200 Ł. Ma˛ ka et al. / Food Control 36 (2014) 199e204
S. Mbandaka (8.79%), replacing S. Typhimurium (8.02%) (Czarkowski,
Cieleba˛ k, Kondej, & Staszewska, 2009; 2011). Based on the Reports on
cases of infectious diseases and poisonings in Poland (http://www.
pzh.gov.pl/oldpage/epimeld/index_p.html#01) can be concluded,
that Salmonella spp. are a major bacterial agent of food poisoning in
Poland.
In the USA, the annual costs resulting from salmonellosis
amount to several billions of dollars (Voetsch et al., 2004). The
European Food Safety Authority (EFSA) has estimated that the
overall economic burden due to human cases in the EU could be as
high as 3 billion euros per year (EFSA, 2011a). The majority of
Salmonella infections are associated with contaminated food:
chicken, pork, dairy products, eggs, fruits, vegetables and others
(Oliveira, Flores, Santos, & Brandelli, 2005; Yan et al., 2010; Zhao
et al., 2008).
Antimicrobial agents are widely use in human and veterinary
medicine, and to improve the growth of animals and plants.
Although the use of antibiotic growth promoters in animal nutri-
tion is now prohibited by EU legislation (Regulation (EC) No. 1831/
2003), some practices in human and animal healthcare, often
resulting from commercial pressure to prescribe and sell antibi-
otics, may lead to inappropriate and overuse of these agents
(Council of the European Union conclusions, 2012). The use of an-
tibiotics in animals, not only to treat disease, but also as growth-
promoting substances and to prevent from diseases, promotes the
development and spread of antimicrobial-resistant bacteria
potentially worldwide (Aminov, 2010). Antibiotics released into the
environment can provoke the formation of resistance, and even
cross- and multiple resistance in bacteria (Allen et al., 2010;
Aminov, 2011; Popowska et al., 2012; Popowska, Miernik,
Rzeczycka, & qopaciuk, 2010). Pathogens as well as commensal
bacteria are affected, the latter constituting a potential reservoir of
resistance genes for pathogenic bacteria. The transfer of such
pathogens through the food chain is possible and consequently
lowers the success of pharmacotherapies for curing humans (Doyle
et al., 2006; EFSA, 2008; Singer et al., 2003).
Resistance genes active against beta-lactams (3rd and 4th
generation) and fluoroquinolones, which are extremely important
antimicrobial agents in human medicine, have been identified on
many livestock farms, and particularly in the poultry industry
where fluoroquinolones are used, so that meat products may be
contaminated with bacterial strains resistant to this antimicrobial
agent. Genes encoding enzymes of the AmpC and Extended
Spectrum Beta-lactamase (ESBL) families are often found in Sal-
monella and E. coli isolates (EFSA, 2011b; Zhao et al., 2008). Ge-
netic analyses of the bacterial strains and resistance genes isolated
from farm animals, foods and human subjects have identified
strong similarities (Leverstein-Van Hall et al., 2011). Such studies
indicate that resistant strains, ESBL genes and mobile genetic el-
ements might be transmitted to humans through the food chain
(Gastmeier, 2010).
The molecular mechanisms of resistance to antibiotics have
been studied extensively and a great deal is now known about the
genetics and biochemistry of many different facets of bacterial cell
function (Alekshun & Levy, 2007; Andersson & Hughes, 2010; Roe
& Pillai, 2003; Walsh & Amyes, 2004). However, it is also very
important to identify any trends in antimicrobial resistance by
conducting regular screening. Most surveys of resistance in Sal-
monella have focused on clinical and veterinary strains. To fully
appreciate the scale of antimicrobial resistance in order to counter
this growing problem, it is also necessary to investigate consumer
exposure to resistant strains present in food. In this study, we have
examined the antimicrobial susceptibility of Salmonella strains
isolated from retail meat products in Poland over the last five
years.
2. Materials and methods
2.1. Strain collection
Between 2008 and 2012, a total of 106 Salmonella strains were
collected by 26 Sanitary and Epidemiological Stations across Poland
in the course of their Official Control and Monitoring Program (18
strains in 2008, 23 in 2009, 22 in 2010, 26 in 2011 and 17 in 2012).
The strains were isolated from retail meat products sampled ac-
cording to PN-EN ISO 6579:2003/A1:2007 “Horizontal method for
the detection of Salmonella spp.”, from poultry meat (n ¼ 81), pork
(n ¼ 7), beef (n ¼ 3) and mixed meat (n ¼ 15) (pork-beef meat).
2.2. Antimicrobial susceptibility testing
The antimicrobial susceptibility of Salmonella isolates was
assessed by tests performed in 2011 and 2012. The antibiotic
resistance profile of each strain was determined using the disk
diffusion method on Mueller-Hinton agar (OXOID, PO5007A), ac-
cording to the methodology of the European Committee on
Antimicrobial Susceptibility Testing e EUCAST (EUCAST, 2012).
Discs containing the following antibiotics (OXOID) were used:
Aztreonam (30 mg), Amoxycillin/clavulanic acid (20/10 mg),
Ampicillin (10 mg), Cefepime (30 mg), Cefotaxime (5 mg), Cefoxitin
(30 mg), Ceftazidime (10 mg), Ceftriaxone (30 mg), Chloramphenicol
(30 mg), Ciprofloxacin (5 mg), Ertapenem (10 mg), Gentamicin
(10 mg), Imipenem (10 mg), Nalidixic acid (30 mg), Sulphonamides
compound (300 mg), Streptomycin (10 mg), Tetracycline (30 mg),
Trimethoprim (5 mg), Trimethoprim/sulphametoxazole (1.25/
23.75 mg).
Individual colonies were suspended in saline to a density of 0.5
on the McFarland turbidity standard, measured using a densi-
tometer (Biomerieux). Each cell suspension was spread over the
entire surface of a plate by swabbing in three directions and the
antibiotic discs were applied. The plates were then incubated at
35  C for 16e20 h. The diameters of the zones of inhibition were
measured and compared with EUCAST (EUCAST, 2012) or Clinical
and Laboratory Standards Institute (CLSI) standards (CLSI, 2012).
In cases where EUCAST breakpoints were absent, the results were
interpreted according to the CLSI breakpoints. Quality control
tests were performed using E. coli ATCC 25922.
2.3. Salmonella serotyping
Isolates were serotyped by Sanitary and Epidemiological Sta-
tions and sent to the Laboratory of Food Microbiology, National
Institute of Public Health e National Institute of Hygiene (NIPHe
NIH), Warsaw, Poland. Where the serotype of a Salmonella isolate
was undefined, serotyping was carried out within the Department
of Bacteriology at the NIPHeNIH. Strains were serotyped by slide
agglutination test with specific O and H antisera (Immunolab,
Biomed) and classified according to the WhiteeKauffmann-Le Mi-
nor scheme (Grimont & Weill, 2007).
3. Results and discussion
3.1. Prevalence of Salmonella serotypes in retail meat products
The Salmonella serotypes isolated from retail meat products of
different animal origin in Poland are listed in Table 1. Most of the
106 Salmonella strains were isolated from poultry meat (n ¼ 81),
while isolates from pork (n ¼ 7), beef (n ¼ 3) and mixed meat
(n ¼ 15) samples were less frequent. A total of twenty-one different
Salmonella serotypes were identified.
 Author's personal copy

Ł. Ma˛ ka et al. / Food Control 36 (2014) 199e204 201

Table 1 Table 2
Incidence of Salmonella serotypes isolated from different retail meats. Incidence of Salmonella serotypes in retail meat products.

Source No. of Serotype % n Salmonella serotype % n

isolates
Salmonella Enteritidis 34.9 37
Poultry 81 Salmonella Enteritidis 34.6 28 Salmonella Infantis 14.2 15
Salmonella Infantis 18.5 15 Salmonella Typhimurium 10.4 11
Salmonella Hadar 8.6 7 Salmonella Hadar 6.6 7
Salmonella Newport 7.4 6 Salmonella Newport 5.7 6
Salmonella 7.4 6 Salmonella Virchow 4.7 5
Typhimurium Salmonella Chester 3.8 4
Salmonella Virchow 6.1 5 Salmonella Agona 2.8 3
Salmonella Saintpaul 3.7 3 Salmonella Saintpaul 2.8 3
Salmonella Chester 2.5 2 Salmonella Derby 1.9 2
Salmonella Duisburg 2.5 2 Salmonella Duisburg 1.9 2
Salmonella Sandiego 2.5 2 Salmonella Sandiego 1.9 2
Salmonella Agona 1.2 1 Salmonella Anatum 0.9 1
Salmonella Anatum 1.2 1 Salmonella Brandenburg 0.9 1
Salmonella Derby 1.2 1 Salmonella Eko 0.9 1
Salmonella Glostrup 1.2 1 Salmonella Glostrup 0.9 1
Salmonella Mbandaka 1.2 1 Salmonella Heidelberg 0.9 1
Pork 7 Salmonella Enteritidis 42.9 3 Salmonella Indiana 0.9 1
Salmonella 14.3 1 Salmonella Kottbus 0.9 1
Typhimurium Salmonella Mbandaka 0.9 1
Salmonella Brandenburg 14.3 1 Salmonella Wippra 0.9 1
Salmonella Agona 14.3 1 Total 106
Salmonella Wippra 14.3 1
Beef 3 Salmonella Enteritidis 66.7 2
Salmonella Indiana 33.3 1
Mixed 15 Salmonella Enteritidis 26.7 4 Yamaguchi, 2012). The findings summarized above indicate that
Salmonella Typhimurium 26.7 4
different Salmonella serotypes are prevalent in separate regions of
Salmonella Chester 13.3 2
Salmonella Agona 6.7 1 the world and that the trade in food has had an impact on world-
Salmonella Eko 6.7 1 wide serotype distribution.
Salmonella Derby 6.7 1
Salmonella Heidelberg 6.7 1 3.2. Antimicrobial susceptibility of Salmonella
Salmonella Kottbus 6.7 1
Total 106
The results of antimicrobial susceptibility tests are shown in
Table 3, Fig. 1 and Fig. 2. Salmonella strains isolated from poultry
meat displayed the widest spectrum of antibiotic resistance (to 12
of 19 tested compounds). Isolates from mixed meat samples
Salmonella Enteritidis was the prevalent serotype in all studied showed the same resistance spectrum as poultry meat products,
types of retail meat product and accounted for 34.9% of all strains. excluding aztreonam, cefoxitin and gentamicin. It is likely that most
The second most common serotype was Salmonella Infantis, which Salmonella isolates from mixed meat originated from poultry. The
comprised 14.2% of all strains. Salmonella Typhimurium was the seven isolates from pork showed resistance to three antimicrobials
third most common serotype. The three Salmonella serotypes most (ampicillin, nalidixic acid and tetracycline), while the three isolates
frequently isolated from retail meat products (S. Enteritidis, S. from beef were resistant to two (nalidixic acid and gentamicin).
Infantis, S. Typhimurium) accounted for 59.4% of all isolates However, due to the small number of isolates from pork and beef it
(Table 2). is hard to draw firm conclusions from this finding. All of the tested
Salmonella Enteritidis and Salmonella Typhimurium, which strains were susceptible to cefepime, cefotaxime, ceftazidime, cef-
comprised 45.3% (n ¼ 48) of all isolates in the present study, are triaxone, ciprofloxacin, ertapenem and imipenem.
also the most frequently isolated Salmonella serotypes in many The most common antibiotic resistance among all Salmonella
countries of Europe, and both are clinically important (EFSA and isolates (52.8% of strains) was to nalidixic acid. Resistance to this
ECDC, 2011). In the UK, S. Typhimurium was the predominant quinolone was observed in strains isolated from all meat sources.
serotype (54.2%) in fresh raw red meat samples from retail outlets This prevalence of resistance to nalidixic acid is in agreement with
or food services (Little, Richardson, Owen, De Pinna, & Threlfall, the findings of previous studies. All tested Salmonella strains iso-
2008). According to Zhao et al. (2008), Salmonella Heidelberg was lated from poultry meat in Spain were resistant to nalidixic acid
the serotype isolated most frequently from retail meats in North (Álvarez-Fernández, Alonso-Calleja, García-Fernández, & Capita,
America. However, Aslam et al. (2012) recently reported that the 2012). In Mexico, higher levels of quinolone resistance were found
most prevalent serotype found in retail chicken meat in Alberta, in isolates from poultry meat compared with those from other
Canada, was S. Hadar, followed by S. Heidelberg and S. Kentucky. In foods (Miranda, Mondragon, Martinez, Guarddon, & Rodriguez,
turkey samples, S. Heidelberg was the most common, followed by S. 2009). Nalidixic acid resistance was most frequent in isolates
Hadar. This study also isolated Salmonella from 2% of pork samples, from chicken meat, but it was also found in the most common
but no strains were detected in ground beef. Bosilevac, Guerini, serotypes (except for S. Derby) isolated from fresh pork sausages
Kalchayanand, and Koohmaraie (2009) reported that the overall (Mürmann, Dos Santos, & Cardoso, 2009; Yan et al., 2010).
incidence of Salmonella isolated from ground beef in the USA was Besides nalidixic acid, the most frequent antibiotic resistance
4.2%. A study of Salmonella strains in retail meat samples from occurring in the Salmonella isolates tested in the present study was
Tehran, Iran, found that S. Thompson was the most prevalent also resistance to tetracycline (32.1%), ampicillin (28.3%), strepto-
serotype followed by S. Hadar (Dallal et al. 2010). In Vietnam, S. mycin (28.3%) and sulphonamides (26.4%). The incidence of resis-
Anatum was the most common serotype found in retail chicken and tance to the aforementioned antimicrobials is similar to that
pork samples, followed by S. Infantis and S. Emek (Thai, Hirai, Lan, & found in strains isolated from raw, chilled, retail chickens in the
 Author's personal copy

202 Ł. Ma˛ ka et al. / Food Control 36 (2014) 199e204

ATM e Aztreonam; AMC e Amoxycillin/clavulanic ac.; AMP e Ampicillin; FEP e Cefepime; CTX e Cefotaxime; FOX e Cefoxitin; CAZ e Ceftazidime; CRO e Ceftriaxone; C e Chloramphenicol; CIP e Ciprofloxacin; ETP e
100(1)
2.7(1)

9.1(1)

50(1)
SXT

0
0
0

0
0
0
0
0
0
0
0
0

0
0
0

0
0
100(1)
2.7(1)

9.1(1)

50(1)
W

0
0
0

0
0
0
0
0
0
0
0
0

0
0
0

0
0

Ertapenem; CN e Gentamicin; IPM e Imipenem; NA e Nalidixic Ac.; SUL e Sulphonamides comp.; STR e Streptomycin; TE e Tetracycline; W e Trimethoprim; SXT e Trimethoprim/sulphametoxazol.
28.6(2)
81.8(9)

33.3(1)

100(1)

100(2)

100(1)

100(1)
100(1)
100(1)
2.7(1)
60(9)

50(3)

50(2)
TE

0
0

0
0

0
71.4(5)
54.5(6)

66.7(2)

100(1)

100(2)

100(1)
2.7(1)
60(9)

50(2)

50(1)
STR

0
0

0
0

0
0
0

0
0
Fig. 1. Antimicrobial susceptibility of Salmonella isolated from various meat sources to
tested antibiotics. ATM e Aztreonam; AMC e Amoxycillin/clavulanic ac.; AMP e
66.7(10)

54.5(6)
33.3(2)

66.7(2)

Ampicillin; FEP e Cefepime; CTX e Cefotaxime; FOX e Cefoxitin; CAZ e Ceftazidime;

100(2)

100(1)

100(1)
2.7(1)

50(2)

50(1)

CRO e Ceftriaxone; C e Chloramphenicol; CIP e Ciprofloxacin; ETP e Ertapenem; CN e

SUL

Gentamicin; IPM e Imipenem; NA e Nalidixic Ac.; SUL e Sulphonamides comp.; STR e

0
0

0
0
0
0

0
0
0

Streptomycin; TE e Tetracycline; W e Trimethoprim; SXT e Trimethoprim/

73.3(11)

sulphametoxazol.
71.4(5)
36.4(4)
54(20)

100(3)

100(1)
100(2)

100(2)

100(1)

100(1)
60(3)

50(3)
NA

0
0

0
0
0
0

UK: sulfonamides (52%), streptomycin (26%), tetracycline (22%) and

IPM

ampicillin (17%) (Wilson, 2004). Among Salmonella strains isolated

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

from poultry and beef products in Iran by Dallal et al. (2010),

resistance to nalidixic acid (82%), tetracycline (69%), trimethoprim
18.2(20)

66.7(2)

100(2)

100(1)

(63%) and streptomycin (52%) was extremely widespread. The

CN

percentage of antibiotic-resistant strains among Iranian isolates

0
0
0
0

0
0

0
0
0
0
0

0
0
0

0
0
0

was far higher than found in the present study, most notably, the
ETP

incidence of resistance to trimethoprim (63% vs. 3.8%). As in Poland,

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

all Salmonella isolates from Iran were sensitive to ciprofloxacin,

imipenem, ceftazidime (Dallal et al., 2010).
CIP

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

Resistance to sulphonamides, streptomycin, tetracycline and

ampicillin has been frequently reported among Salmonella isolates
45.4(5)

33.3(1)

100(1)
6.7(1)

from retail meats sampled in East Asia, but unlike the present study,
the tested strains were often also resistant to chloramphenicol
C

0
0

0
0

0
0
0
0
0
0
0
0
0
0
0

(37.3e42.1%), ciprofloxacin (42.1%) and trimethoprim (34.0%) (Thai

CRO

et al., 2012; Yan et al., 2010). None of the Salmonella strains isolated
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

in North Vietnam by Thai et al. (2012) were resistant to ceftazidime.

In a study examining Salmonella in raw red meats in the UK, the
CAZ

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

most frequent antimicrobial resistance found in isolates was to

tetracyclines (67.5%), sulphonamides (56.6%), streptomycin (50.6%),
20 (1)
6.7(1)
FOX

ampicillin (43.4%) and chloramphenicol (33.7%) (Little et al., 2008).

0

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

In Austria, the highest resistance rate among Salmonella isolated

from meat samples was seen for nalidixic acid (42%), followed by
CTX

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
Antimicrobial resistance among different serotypes of Salmonella % (n).

FEP

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
71.4(5)
72.7(8)
83.3(5)

100(3)

100(2)

100(1)

100(1)

100(1)
2.7(1)
6.7(1)
20(1)

50(1)
AMP

0
0
0
0
0

0
36.4(4)
16.7(1)

100(3)

100(2)

100(1)
6.7(1)
20(1)
57(4)
AMC
Antimicrobial

0
0
0
0
0

0
0
0
0
0

0
16.7(1)
ATM

0
0
0
0
0

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
isolates
No. of

37
15
5
7
11
6
3
3
1
1
1
2
4
2
2
1
1
1
1
1
1

Fig. 2. Percentage of Salmonella isolates resistant to the tested antibiotics. ATM e

Typhimurium

Brandenburg

Aztreonam; AMC e Amoxycillin/clavulanic ac.; AMP e Ampicillin; FEP e Cefepime;

Heidelberg
Mbandaka
Enteritidis

Saintpaul

Duisburg

Sandiego
Newport
Serotype

Glostrup
Virchow

CTX e Cefotaxime; FOX e Cefoxitin; CAZ e Ceftazidime; CRO e Ceftriaxone; C e

Anatum

Kottbus
Chester
Infantis

Wippra
Indiana
Agona
Hadar

Derby

Chloramphenicol; CIP e Ciprofloxacin; ETP e Ertapenem; CN e Gentamicin; IPM e

Table 3

Eko

Imipenem; NA e Nalidixic Ac.; SUL e Sulphonamides comp.; STR e Streptomycin; TE e

Tetracycline; W e Trimethoprim; SXT e Trimethoprim/sulphametoxazol.
 Author's personal copy

Ł. Ma˛ ka et al. / Food Control 36 (2014) 199e204 203

tetracycline (almost 33%), streptomycin (27%), ampicillin and Table 4

chloramphenicol (both 17%), and ciprofloxacin (9.6%) (Mayrhofer, Multiresistance in various Salmonella serotypes.

Paulsen, Smulders, & Friederike, 2004). Serotype Number of antimicrobials

The results of the present study support the notion that the 1 2 3 4 5 6 7 8
frequency of antimicrobial resistance is higher among Salmonella
Salmonella Enteritidis 19 1
strains isolated from poultry meat than in those from other meat
Salmonella Infantis 1 1 1 8 1
sources, e.g. pork (Thai et al., 2012). Salmonella Typhimurium 2 1 1 1 1 2 2
Examination of the resistance profiles of individual Salmonella Salmonella Hadar 1 1 3 1
serotypes (Table 3) showed that among Salmonella Enteritidis iso- Salmonella Virchow 3 1
Salmonella Newport 2 1 2 1
lates, 54% (n ¼ 20) were resistant to nalidixic acid and one was
Salmonella Saintpaul 1 1
multiresistant (AMP, NA, SUL, STR, TE, W, SXT). Of the Salmonella Salmonella Eko 1
Infantis strains, 73.3% (n ¼ 11), were resistant to nalidixic acid, Salmonella Derby 1
66.7% (n ¼ 10) were resistant to sulphonamides, while 60% (n ¼ 9) Salmonella Heidelberg 1
showed resistance to both streptomycin and tetracycline. Salmo- Salmonella Glostrup 1
Salmonella Wippra 1
nella Typhimurium displayed resistance to the widest spectrum of
Salmonella Indiana 1
antimicrobials: 10 of the 19 tested compounds. Among these Salmonella Duisburg 2
strains, resistance to tetracycline was most prevalent (81.8%, n ¼ 9) Salmonella Kottbus 1
and many strains were resistant to ampicillin (72.7%, n ¼ 8), and Salmonella Chester 2
Salmonella Sandiego 2
sulphonamides and streptomycin (54.5%, n ¼ 6). Of all the isolates,
Total: 31 4 10 13 6 6 3
only one strain of Salmonella Newport was resistant to aztreonam.

Our findings are similar to those of previous studies with respect

3.3. Multiresistance
Of the 106 Salmonella strains isolated from meat samples in the
present study, 68.9% (n ¼ 73) displayed antibiotic resistance. The
frequency of resistance among isolates was high and varied in
particular years between 59.1% in 2010 and 84.6% in 2011 (Fig. 3).
Overall, 42.5% (n ¼ 31) of the resistant isolates showed resistance to
one antibiotic, 5.5% (n ¼ 4) to two, 13.7% (n ¼ 10) to three, 17.8%
(n ¼ 13) to four, and 20.5% (n ¼ 15) were resistant to five or more
antibiotics (Table 4).
Among the 37 Salmonella Enteritidis isolates, 54% (n ¼ 20) dis-
played antibiotic resistance. A higher percentage of resistant iso-
lates was found in some of the less frequently identified serotypes:
Salmonella Newport (100%, i.e. 6 resistant strains/6 isolates), Sal-
monella Typhimurium (91%, 10/11), Salmonella Hadar (85.7%, 6/7),
Salmonella Virchow (80%, 4/5) and Salmonella Infantis (80% 12/15).
Of the 20 resistant S. Enteritidis isolates, 19 strains were resis-
tant to one antibiotic (nalidixic acid) and one was resistant to 7
antibiotics. Multiresistance (resistance to 3 or more antimicrobials)
was seen in a higher proportion of some less frequently identified
serotypes: Salmonella Newport (3 multiresistant strains among 6
resistant isolates), Salmonella Typhimurium (7/10), Salmonella
Hadar (5/6), Salmonella Infantis (10/12). The percentage of multi-
resistant strains among antibiotic resistant isolates varied between
23.1% in 2010 and 81.8% in 2012. The greatest differences was
observed between the year and the frequency of multiresistant
isolates for the scope of years 2010e2012 (Fig. 3).
Fig. 3. Percentage of resistant and multiresistant Salmonella isolates by year.
to the antibiotic resistance rate in particular Salmonella serotypes:
resistance has been observed more frequently in S. Typhimurium, S.
Infantis and S. Virchow than in S. Enteritidis (Ma˛ ka et al., 2010; Thai
et al., 2012; Threlfall, Ward, Frost, & Willshaw, 2000). In Turkey, 62%
of Salmonella strains isolated from meat were resistant to three or
more antimicrobial agents (Arslan & Eyi, 2010). In Spain, the
average number of antibiotic resistance determinants per strain
was lowest in isolates of Salmonella Enteritidis (4.64), while higher
values were seen in S. Infantis (5.5), S. Newport (6.5) and S.
Typhimurium (6.0) (Álvarez-Fernández et al., 2012). Among strains
isolated from meat samples in Austria, 36% of S. Enteritidis isolates
were resistant to at least one of the antimicrobials tested, all four S.
Virchow isolates showed resistance, one of the three isolates of S.
Typhimurium was sensitive to all tested antimicrobials, and the
single S. Hadar isolate was multiresistant (Mayrhofer et al., 2004).
Multiresistance has become a significant public health problem.
Antibiotic resistance is a factor that can influence the severity of
salmonellosis. Moreover, infections caused by multiresistant bac-
teria are often much more difficult to treat because the panel of
effective antimicrobials is reduced and antimicrobial therapy can
be delayed or inadequate (Magiorakos et al., 2012; Roberts et al.,
2009).
4. Conclusions
This study examined Salmonella strains isolated in Poland be-
tween 2008 and 2012 from retail meat products; mostly from
poultry, but also from beef, pork and mixed meat. The serotype
found most often was Salmonella Enteritidis. Salmonella strains
isolated from poultry products were resistant to a wider spectrum
of antimicrobials that those of other origins. Among the isolates,
resistance to nalidixic acid was prevalent, but tetracycline, ampi-
cillin, streptomycin and sulphonamides resistance was also com-
mon. Multiresistance was more frequently observed among isolates
of S. Newport, S. Typhimurium, S. Hadar, S. Virchow and S. Infantis,
than in the more abundant S. Enteritidis strains.
Although the number of Salmonella strains isolated from retail
meats in Poland decreased over the 5-year study period, other
potential problems remain. In particular, levels of antibiotic resis-
tance and multiresistance are high among the isolated strains. This
represents an additional risk for consumers because therapy be-
comes more complicated in the case of illness. Even where the
consumption of contaminated food does not cause illness, the
 Author's personal copy
204 Ł. Ma˛ ka et al. / Food Control 36 (2014) 199e204
introduction of antibiotic resistant bacterial strains can lead to the
transfer of resistance genes to the host bacteria via HGT. In view of
the observed increase in the resistance level of Salmonella strains,
regular surveillance is essential for the early detection of any shift
in the antimicrobial resistance profile of isolates from food and the
environment. Our findings indicate that food is a potential source of
resistant bacteria that are a serious concern for public health and
food safety.
Acknowledgments
This work was supported by a grant from the National Center of
Science DEC-2011/01/N/NZ9/00197 and partly supported by a grant
from the State Committee for Scientific Research, Poland (741/N-
COST/2010/0). The research was conducted as part of European Co-
operation in the field of Scientific and Technical (COST) Research
Action TD0803 “Detecting evolutionary hot spots of antibiotic re-
sistances in Europe (DARE)” (2009e2013).
The authors would like to thank Dr. Jolanta Szych, Monika
Wasiak and the Department of Bacteriology of the National Insti-
tute of Public Health e National Institute of Hygiene (Warsaw,
Poland) for serotyping the Salmonella isolates.
References
Alekshun, M. N., & Levy, S. B. (2007). Molecular mechanisms of antibacterial
multidrug resistance. Cell, 128, 1037e1050.
Allen, H. K., Donato, J., Wang, H. H., Cloud-Hansen, K. A., Davies, J., & Handelsman, J.
(2010). Call of the wild: antibiotic resistance genes in natural environments.
Nature Reviews Microbiology, 8, 251e259.
Álvarez-Fernández, E., Alonso-Calleja, C., García-Fernández, C., & Capita, R. (2012).
Prevalence and antimicrobial resistance of Salmonella serotypes isolated from
poultry in Spain: comparison between 1993 and 2006. International Journal of
Food Microbiology, 153(3), 281e287.
Aminov, R. I. (2010). A brief history of the antibiotic era: lessons learned and
challenges for the future. Frontiers in Microbiology, 1, 134.
Aminov, R. I. (2011). Horizontal gene exchange in environmental microbiota.
Frontiers in Microbiology, 2, 158.
Andersson, D. I., & Hughes, D. (2010). Antibiotic resistance and its cost: is it possible
to reverse resistance? Nature Reviews Microbiology, 8(4), 260e271.
Arslan, S., & Eyi, A. (2010). Occurrence and antimicrobial resistance profiles of
Salmonella species in retail meat products. Journal of Food Protection, 73, 1613e
1617.
Aslam, M., Checkley, S., Avery, B., Chalmers, G., Bohaychuk, V., Gensler, G., et al.
(2012). Phenotypic and genetic characterization of antimicrobial resistance in
Salmonella serovars isolated from retail meats in Alberta, Canada. Food Micro-
biology, 32(1), 110e117.
Bosilevac, J. M., Guerini, M. N., Kalchayanand, N., & Koohmaraie, M. (2009). Preva-
lence and characterization of Salmonellae in commercial ground beef in the
United States. Applied and Environmental Microbiology, 75(7), 1892e1900.
CLSI. Clinical and Laboratory Standards Institute. (2012). Supplement M100-S22.
Performance standards for antimicrobial susceptibility testing; twenty-second in-
formation (Vol. 32) (3). Wayne, PA.
Council conclusions of 22 June 2012 on the impact of antimicrobial resistance in the
human health sector and in the veterinary sector e a “One Health” perspective.
Official Journal of the European Union C211, 55, 2e5.
Czarkowski, M. P., Cieleba˛ k, E., Kondej, B., & Staszewska, E. (2009). Infectious diseases
and poisonings in Poland in 2008. Warsaw, Poland.
Czarkowski, M. P., Cieleba˛ k, E., Kondej, B., & Staszewska, E. (2011). Infectious diseases
and poisonings in Poland in 2010. Warsaw, Poland.
Dallal, M. M. S., Doyle, M. P., Rezadehbashi, M., Dabiri, H., Sanaei, M., Modarresi, S.,
et al. (2010). Prevalence and antimicrobial resistance profiles of Salmonella
serotypes, Campylobacter and Yersinia spp. isolated from retail chicken and beef,
Tehran, Iran. Food Control, 21(4), 388e392.
Doyle, M. P., Busta, F., Cords, B. R., Davidson, P. M., Hawke, J., Hurd, H. S., et al. (2006).
Antimicrobial resistance: implications for the food system e an expert report,
funded by the IFT Foundation. Comprehensive Reviews in Food Science and Food
Safety, 5, 71e137.
EFSA. (2008). Scientific opinion of the Panel on Biological Hazards on a request from
the European Food Safety Authority on foodborne antimicrobial resistance as a
biological hazard. The EFSA Journal, 765, 1e87.
EFSA (2011a). http://www.efsa.europa.eu/en/corporate/doc/factsheetsalmonella.pdf.
EFSA, & EFSA Panel on Biological Hazards (BIOHAZ).. (2011b). Scientific opinion on
the public health risks of bacterial strains producing extended-spectrum b-
lactamases and/or AmpC b-lactamases in food and food-producing animals. The
EFSA Journal, 9(8), 1e95, 2322.
EFSA and ECDC. (2011). The European Union summary report on trends and sources
of zoonoses, zoonotic agents and food-borne outbreaks in 2009. EFSA Journal,
9(3), 2090.
EUCAST (the European Committee on Antimicrobial Susceptibility Testing).
(February 2012). Antimicrobial susceptibility testing. EUCAST disk diffusion
method, Version 2.1.
Gastmeier, P. (2010). Healthcare-associated versus community-acquired infections:
a new challenge for science and society. International Journal of Medical
Microbiology, 300(6), 342e345.
Grimont, P. A. D., & Weill, F.-X. (2007). Antigenic formulae of the Salmonella serovars”
(9th ed.). WHO Collaborating Centre for Reference and Research on Salmonella,
Institut Pasteur.
Guibourdenche, M., Roggentin, P., Mikoleit, M., Fields, P. I., Bockemühl, J.,
Grimont, P. A. D., et al. (2010). Supplement 2003e2007 (No. 47) to the White-
Kauffmann-Le Minor scheme. Research in Microbiology, 161(1).
Leverstein-Van Hall, M. A., Dierikx, C. M., Stuart, J. C., Voets, G. M., Van Den
Munckhof, M. P., Van Essen-Zandbergen, A., et al. (2011). Dutch patients, retail
chicken meat and poultry share the same ESBL genes, plasmids and strains.
Clinical Microbiology and Infection, 17, 873e880.
Little, C. L., Richardson, J. F., Owen, R. J., De Pinna, E., & Threlfall, E. J. (2008).
Campylobacter and Salmonella in raw red meats in the United Kingdom: prev-
alence, characterization and antimicrobial resistance pattern, 2003e2005. Food
Microbiology, 25(3), 538e543.
Magiorakos, A.-P., Srinivasan, A., Carey, R. B., Carmeli, Y., Falagas, M. E., Giske, C. G.,
et al. (2012). Multidrug-resistant, extensively drug-resistant and pandrug-
resistant bacteria: an international expert proposal for interim standard
definitions for acquired resistance. Clinical Microbiology and Infection, 18,
268e281.
 zy
Ma˛ ka, q., Scie _ n
 ska, H., Grochowska, A., Paw1owska, K., Windyga, B., &
Kar1owski, K. (2010). Antimicrobial susceptibility of Salmonella strains isolated
from food in Poland. Bromatologia i Chemia Toksykologiczna, 43(3), 260e265.
Mayrhofer, S., Paulsen, P., Smulders, F. J. M., & Friederike, H. (2004). Antimicrobial
resistance profile of five major food-borne pathogens isolated from beef, pork
and poultry. International Journal of Food Microbiology, 97, 23e29.
Miranda, J. M., Mondragon, A. C., Martinez, B., Guarddon, M., & Rodriguez, J. A.
(2009). Prevalence and antimicrobial resistance patterns of Salmonella from
different raw foods in Mexico. Journal of Food Protection, 72, 966e971.
Mürmann, L., Dos Santos, M. C., & Cardoso, M. (2009). Prevalence, genetic charac-
terization and antimicrobial resistance of Salmonella isolated from fresh pork
sausages in Porto Alegre, Brazil. Food Control, 20(3), 191e195.
Oliveira, D. S., Flores, F. S., Santos, L. R., & Brandelli, A. (2005). Antimicrobial resis-
tance in Salmonella Enteritidis strains isolated from broiler carcasses, food,
human and poultry-related samples. International Journal of Food Microbiology,
97, 297e305.
Popowska, M., Rzeczycka, M., Miernik, A., Krawczyk-Balska, A., Walsh, F., & Duffy, B.
(2012). Influence of soil use on prevalence of tetracycline, streptomycin, and
erythromycin resistance and associated resistance genes. Antimicrobial Agents
and Chemotherapy, 56(3), 1434e1443.
Popowska, M., Miernik, A., Rzeczycka, M., & qopaciuk, A. (2010). The impact of
environmental contamination with antibiotics on levels of resistance in soil
bacteria. Journal of Environmental Quality, 39(5), 1679e1687.
Regulation (EC) No 1831/2003 of the European Parliament and of the Council of 22
September 2003 on additives for use in animal nutrition.
Roberts, R. R., Hota, B., Ahmad, I., Scott, D., II., Foster, S. D., Abbasi, F., et al. (2009).
Hospital and societal costs of antimicrobial-resistant infections in a Chicago
teaching hospital: implications for antibiotic stewardship. Clinical Infectious
Diseases, 49(8), 1175e1184.
Roe, M. T., & Pillai, S. D. (2003). Monitoring and identifying antibiotic resistance
mechanisms in bacteria. Poultry Science, 82, 622e626.
Singer, R. S., Finch, R., Wegener, H. C., Bywater, R., Walters, J., & Lipsitch, M. (2003).
Antibiotic resistance e the interplay between antibiotic use in animals and
human beings. The Lancet Infectious Diseases, 3(1), 47e51.
Thai, T. H., Hirai, T., Lan, N. T., & Yamaguchi, R. (2012). Antibiotic resistance profiles
of Salmonella serovars isolated from retail pork and chicken meat in North
Vietnam. International Journal of Food Microbiology, 156(2), 147e151.
Threlfall, E. J., Ward, L. R., Frost, J. A., & Willshaw, G. A. (2000). The emergence and
spread of antibiotic resistance in food-borne bacteria. International Journal of
Food Microbiology, 62, 1e5.
Voetsch, A. C., Van Gilder, T. J., Angulo, F. J., Farley, M. M., Shallow, S., Marcus, R.,
et al. (2004). FoodNet estimate of the burden of illness caused by non typhoidal
Salmonella infections in the United States. Clinical Infectious Diseases,
38(Suppl. 3), S127eS134.
Walsh, F. M., & Amyes, S. G. B. (2004). Microbiology and drug resistance mech-
anisms of fully resistant pathogens. Current Opinion in Microbiology, 7(5),
439e444.
Wilson, I. G. (2004). Antimicrobial resistance of Salmonella in retail chickens, im-
ported chicken portions, and human clinical specimens. Journal of Food Pro-
tection, 67, 1220e1225.
Yan, H., Li, L., Alam, M. J., Shinoda, S., Miyoshi, S.-I., & Shi, L. (2010). Prevalence and
antimicrobial resistance of Salmonella in retail foods in northern China. Inter-
national Journal of Food Microbiology, 143(3), 230e234.
Zhao, S., White, D. G., Friedman, S. L., Glenn, A., Blickenstaff, K., Ayers, S. L., et al.
(2008). Antimicrobial resistance in Salmonella enterica serovar Heidelberg iso-
lates from retail meats, including poultry, from 2002 to 2006. Applied and
Environmental Microbiology, 74, 6656e6662.