Salmonella spp.
A Wingstrand and S Aabo, Technical University of Denmark, Søborg, Denmark
r 2014 Elsevier Ltd. All rights reserved.
Glossary
Enzyme-linked immunosorbent assay (ELISA) method A
detection method in which antigens or antibodies are fixed
in wells. Antigen−antibody complexes are formed after
addition of a sample containing the corresponding
antibody or antigen. Complexes are labeled with enzymes
and visualized by an enzymatic color reaction.
Hazard analysis critical control point (HACCP)
systems A scientific approach to establishing plant- or
process-specific control systems in the food industry.
Host-adapted The adaptation of a microorganism to a
specific host species. Therefore, the microorganism is found
mostly in the host species and only infrequent in other
species.
Lipopolysaccharides (LPS) An antigenic part of the outer
cell membrane of Gram-negative bacteria.
Introduction
Salmonella is a pathogen, the main reservoir of which is the
gastrointestinal tract of warm-blooded animals. Among nu-
merous serovars of Salmonella enterica subsp. enterica (S.),
serovar Typhi and the serovars Paratyphi A and B infect only
humans and are not spread from animals. The remaining
(nontyphoid) Salmonella serovars originate from the animal
reservoir. Through fecal contamination of meat during
slaughter, meat animals are among the most important sour-
ces of human salmonellosis.
The effort to reduce human salmonellosis is challenged by
a widespread, mostly subclinical occurrence of Salmonella in a
variety of meat animals together with an ability of Salmonella
to adapt to and survive changing environmental conditions.
Since the early 1990s, many new detection and typing
methods, surveillance programs, and control methods for
Salmonella in the farm-to-fork continuum have become avail-
able and have been implemented in an increasing number of
countries worldwide. The growing awareness of a global food
market has led to international initiatives toward global con-
trol of Salmonella.
Characteristics of Salmonella
The genus Salmonella belongs to the family Enterobacteriaceae.
Salmonella are Gram-negative, facultatively anaerobic, motile
rods that are catalase positive and cytochrome oxidase nega-
tive, produce gas from glucose, and are able to reduce nitrate.
Salmonella and Escherichia coli are closely related and are be-
lieved to share a common ancestor. During evolution E. coli
has acquired the ability to utilize lactose through being closely
Encyclopedia of Meat Sciences, Volume 2 doi:10.1016/B978-0-12-384731-7.00034-9
Macrophage A cell that is specialized for engulfing foreign
material and microorganisms that gain access to the body.
Macrophages break down the foreign substances and
present them to the immune system.
Muscle fluid The fluid obtained from thawed meat (also
called meat juice).
Salmonellosis A general term for clinical disease caused
by Salmonella, but frequently restricted to clinical infections
from nontyphoid Salmonella serovars.
Sequelae The secondary clinical conditions following a
primary disease.
Subclinical infection An infection without recognizable
clinical symptoms.
Zoonoses The diseases that can be transmitted between
animals and humans.
associated with mammals, whereas Salmonella is unable to
utilize lactose and is more associated with reptiles and
birds. The acquisition of pathogenicity islands, of which SPI-1
and SPI-2 are the most prominent, conferred virulence on
Salmonella.
The genus Salmonella is comprised of two species: Salmon-
ella enterica and Salmonella bongori. Salmonella enterica is com-
posed of more than 2500 serovars, but Salmonella bongori has
only 22 serovars. Salmonella enterica is divided into 6 sub-
species. Subsp. I (enterica) is comprised of approximately 1500
serovars and is particularly associated with infections in warm-
blooded animals (Table 1).
Seroagglutination of outer membrane O-antigens (LPS)
and flagella H-antigens define the serovar according to the
Kaufmann−White scheme. For example the antigenic structure
O:1,4,5,12 and H1/H2 b:1,2 defines Salmonella enterica subsp.
enterica serovar Typhimurium (in short Salmonella Typhimur-
ium or S. Typhimurium).
A few serovars are termed host specific or host adapted.
Salmonella Typhi, S. Paratyphi A, S. Paratyphi C and S. Sendai
infect only humans. Salmonella Choleraesuis is associated with
pigs and S. Dublin is adapted to cattle but both serovars can
cause serious infections in humans. Salmonella Gallinarum and
S. Pullorum are specific for poultry. The factors that determine
host specificity have not been clarified. In general, the re-
maining serovars of subspecies 1 are zoonotic and have a
wider host range. The two most prominent serovars in human
disease are S. Typhimurium, which in particular has a broad
host spectrum, and S. Enteritidis, which can infect many hosts
but has a predilection for poultry.
In general Salmonella grow between 5 °C and 46 °C with
growth being optimal at temperatures between 35 °C and 37 °C.
Physical conditions such as temperature, salinity, pH, and water
367
368 Microbiological Safety of Meat | Salmonella spp.
Table 1 Taxonomy of the genus Salmonellaa and top five serovars of S. enterica subsp. enterica reported to World Health Organization, Global
Foodborne Infections Network (GFN), 2009. Isolates reported to GFN are dominated by reports from Europe
Genus Species Subspecies (subspecies number)
Salmonella S. enterica S. enterica subsp. enterica (I)
S. enterica subsp.
S. enterica subsp.
S. enterica subsp.
S. enterica subsp.
S. enterica subsp.
S. bongori
a
Grimont, P.A.D., Weill, F.-X., 2007. Antigenic formulae of the Salmonella serovars. 9th ed., WHO Collaborating Center for Reference and Research on Salmonella. France:
Institut Pasteur.
b
World Health Organization, Global Foodborne Infections Network. Country Databank. Available at: http://thor.dfvf.dk/portal/page?_pageid=53,1&_dad=portal&_schema=
PORTAL (accessed 30.03.12).
activity will affect the growth rate. Salmonella can grow between
pH 4.5 and pH 9.0 with optimal growth at pH 6.5 to 7.5.
Salmonella do not grow at water activities below 0.93. Although
the generation time for Salmonella is rather long at low tem-
peratures, significant growth in fresh meat can occur at tem-
peratures above 5 °C, which may pose a consumer risk.
Although the background flora in meat can be numerous and
interactions with the meat flora can reduce the growth rate of
Salmonella, they will not stop Salmonella growing. A range of
Salmonella serovars have been shown to survive freezing for
months without any substantial reductions in numbers.
Isolation and Identification of Salmonella
Conventional Culture Detection
Several media have been developed for culture and isolation of
Salmonella from food. Owing to low numbers of Salmonella in
meat, direct plating of samples on selective agars lacks sensitiv-
ity. Salmonella detection in foods requires three culturing steps:
1. Preenrichment, to allow recovery and growth of injured
and uninjured cells. Typical preenrichment media are buf-
fered peptone water (BPW) and lactose broth (LB).
2. Selective enrichment in a broth medium that suppresses
growth of most bacteria but supports growth of Salmonella.
Typical media are Rappaport−Vassiliadis broth (RV), sel-
enite cystine broth (SC), or tetrathionate broth (TB). From
the RV medium a modified semisolid agar has been de-
veloped (MSRV), which allows detection due to swarming
of motile Salmonella.
3. Plating on indicative media. The indicative media take
advantage of biochemical features such as the ability to
grow in the presence of bile salts and fermentation of su-
crose or xylose, but not lactose. Examples are brilliant green
agar (BGA); bismuth sulfite agar (BSA), and xylose lysine
deoxycholate agar (XLD).
Number of serovars Global top five serovars reported in
humans 2009b (% of isolates)
1531 S. Enteritidis (69.0)
S. Typhimurium (14.0)
S. Infantis (5.7)
S. Virchow (1.3)
S. Newport (1.0)
salamae (II) 505
arizonae (IIIa) 99
diarizonae (IIIb) 336
houtenae (IV) 73
indica (VI) 15
22
Suspect colonies are subcultured on nonselective media
that allow seroagglutination and verification of the serovar.
International standardization committees such as the Inter-
national Standards Organization (ISO), the American Associ-
ation of Analytical Chemists (AOAC), the International Dairy
Federation (IDF), and the Nordic Committee on Food Analysis
(NMKL) provide procedures for Salmonella detection to sup-
port international harmonization.
Rapid Detection Methods
As culture detection of Salmonella requires 3–5 days to provide
a positive result, rapid detection methods have been de-
veloped. They are typically DNA based, e.g., polymerase chain
reaction (PCR), or immunological- (antibody) based methods,
e.g., enzyme-linked immunosorbent assay (ELISA). Although
termed rapid, preenrichment is often necessary to obtain the
required numbers of Salmonella, 104–105 cells ml−1, needed
for detection. These methods have reduced detection times to
between 12 h and 24 h, which enable release of meat for
shipment direct from cooling facilities at the slaughterhouse.
Immunological methods are developed based on inter-
actions between Salmonella antigens and specific antibodies
raised against Salmonella. The format can vary. The linking of
an enzyme to the antibodies allows antigen−antibody com-
plexes to be detected by conversion of a substrate of the
enzyme with development of a visible color, light, or fluor-
escence when Salmonella is present.
For DNA-based methods, PCR methods are predominant.
The principle of PCR is an enzymatic-driven multiplication of
a Salmonella-specific portion of the Salmonella genome. The
reaction is exponential and can produce a positive result in less
than 2 h. However, the reaction takes place in a very small
volume and a preenrichment step is usually needed to obtain
sufficient cells from which the required amount of DNA (or
RNA when relevant) can be extracted. With the recent devel-
opment of real-time PCR, a robust and sensitive DNA-based
method of detection has been made available.
 Microbiological Safety of Meat | Salmonella spp. 369

Typing Methods for Epidemiology and Outbreak Investigation
Phage typing
Phage typing of Salmonella is based on the ability of specific
bacterial viruses, i.e., bacteriophages, to destroy Salmonella. In
phage typing, a number of bacteriophages are spotted onto
agar plates with a confluent culture of the isolate to be typed. If
a phage is able to infect and destroy the isolate, a spot cleared
of cells appears. The pattern of spots determines the phage
type. A phage typing scheme has been developed for a range of
Salmonella serovars, and it has been part of the classical char-
acterization for S. Enteritidis and S. Typhimurium for half a
century (Figure 1).
prominent in Salmonella epidemiological and outbreak
investigation.
Multiple loci variable number of tandem repeats analysis
(MLVA)
MLVA typing is based on the occurrence of short, repetitive
base sequences in the DNA of the chromosome. The numbers
of sequences vary, and strains with an identical number of
sequences are considered identical. Each of five regions of
the chromosome is analyzed for the number of repetitive
sequences. The method has become important for typing of
Salmonella (Figure 1).

Detection of Antibodies to Salmonella by Enzyme

Pulsed-field gel electrophoresis (PFGE)
In these methods, the bacterial chromosome is cut into large
fragments by restriction enzymes and the fragments are sep-
arated by electrophoresis in a pulsing electric field. Bacteria
showing the same bands on the gel are considered to be the
same. PFGE has been widely used and has long been the
standard method for investigation of Salmonella outbreaks
(Figure 1). Other novel technologies are now replacing PFGE.
These include multilocus sequence typing, multiple loci vari-
able number of tandem repeats analysis and full genome
sequencing.
Multilocus sequence typing (MLST)
MLST is based on sequencing of internal fragments of seven
housekeeping genes. Different base sequences (alleles) can
occur in each of the seven genes. Sequences for all seven genes
are stored in an international database. To obtain a MLST type,
the seven gene sequences determined for an isolate are for-
warded to the database and a sequence type is returned.
If sequences for two isolates are identical they will be classified
as the same MLST type. MLST is becoming increasingly
Immunoassay
Since the mid-1990s detection of specific antibodies to Sal-
monella LPS in blood samples, muscle fluid, milk, or egg yolk
from food animals has been possible. Despite the inherent
delay in antibody response (1–2 weeks after infection), an
association between seropositivity and shedding of Salmonella
has been documented. Serology is a convenient, inexpensive
and sensitive method for monitoring and classification of
Salmonella infection in groups of animals, but it is less
suitable for testing individual animals. ELISA test kits are
commercially available. Serological assays tailored to targeted
Salmonella serovars are parts of Salmonella surveillance pro-
grams and are used for research worldwide.
Characteristics of Salmonellosis
Salmonellosis in Meat Animals
Most Salmonella infections in meat animals are subclinical;
but infections with the host-adapted Salmonella serovars can

Lane Comparison of MLVA profile based on PFGE profile MLVA Serotype Source 1 Phage Source 2
MLVA profiles number of repeated units Type – XbaI profile type type
100
70
60

80

90

Loci STTR 9-5-6-10-3

1 4 12 7 9 211 A JPX.0158.DK Typhimurium Human U288

2 4 13 7 9 211 A JPX.0936.DK Typhimurium Pig U288 Colon contents
3 4 13 8 9 211 A JPX.0926.DK Typhimurium Cattle U288 Isolate from fresh beef
4 4 16 9 9 211 B JPX.0995.DK Typhimurium Human U288
5 4 16 9 9 211 B JPX.0995.DK Typhimurium Human U288
6 4 16 9 9 211 B JPX.0995.DK Typhimurium Human U288
7 4 16 9 9 211 B JPX.0995.DK Typhimurium Pig U288 Isolate from food
8 4 16 9 9 211 JPX.0995.DK Typhimurium Pig U288 Pen sample
9 4 13 9 9 211 A JPX.0533DK Typhimurium Pig U288 Isolate from animal
10 4 14 11 9 211 A JPX.0840.DK Typhimurium Human U288
11 4 14 11 9 211 A JPX.0840.DK Typhimurium Human U288
12 4 14 10 9 211 C JPX.0845.DK Typhimurium Human U288
13 4 14 10 9 211 A JPX.0845.DK Typhimurium Human U288
14 4 14 9 10 211 A JPX.0167.DK Typhimurium Pig U288 Isolate from animal

Figure 1 Subtyping methods applied to a selection of five S. Typhimurium isolates from an investigation of a human outbreak of S. Typhimurium
phage type U288 in Denmark, Norway, and Sweden in 2008a (lane 4–8). All had identical MLVA type (JPX.0995.DK), and four were PFGE-typed
and had identical PFGE types (type B). Lane 4–6: Three human outbreak isolates from Danish cases. Lane 7: Food isolate from Danish raw pork
sausage meant for heat treatment. Lane 8: Isolate from a pen fecal sample from a Danish sow herd. The two food and animal isolates of MLVA
JPX.0995.DK were epidemiologically related to a specific pig slaughterhouse and cutting plant identified as the site of contamination. Isolates in
lane 1–3 and 9–14 are added for comparison and were not related to the outbreak. Source 1: Animal species or human origin. Source 2: Sample
type of nonhuman isolates. Printed with permission from Sørensen, G., Diagnostic Engineering, Division of Food Microbiology, National Food
Institute, Technical University of Denmark. aBruun, T., Sørensen, G., Forshell, L.P., et al., 2009. An outbreak of Salmonella Typhimurium infections
in Denmark, Norway and Sweden, 2008. Eurosurveillance 14, 1–6.
370 Microbiological Safety of Meat | Salmonella spp.

cause severe, acute, clinical diseases. Outbreaks of such
diseases can have high mortality rates and cause heavy
economic losses. Symptoms are mainly related to septicemia
(fever, weakness, loss of appetite), but enteritis is also
common, and pneumonia, reproductive failure, and abor-
tion may occur.
Non host-adapted serovars of Salmonella are considered
potentially capable of infecting most meat animals, but certain
serovars are more commonly isolated from certain meat ani-
mal species (e.g., S. Enteritidis in poultry, S. Derby in pigs)
(Table 2). Infection of meat animals with the non host-
adapted serovars may occasionally cause herd outbreaks of
mainly gastrointestinal infections (diarrhea, fever, and de-
hydration) in young animals. Compared to infections with
host-adapted serovars, infections with non host-adapted ser-
ovars generally have a lower mortality.

Table 2 Annual incidence of notified human salmonellosis (confirmed cases) in European Union (EU) Member States, 2010, and detection of
Salmonella in broilers, turkeys, and slaughter pigs in baseline studies conducted in EU Member States between 2005 and 2007. Top five
Salmonella serovars from humans and meat animals are listed below

EU member state Human salmonellosis Salmonella detected by culture in EU baseline studies

2010a,b (cases per 100 000)
Broilers 2005−2006e Turkeys 2006−2007f Slaughter pigs 2006−2007g
(% of flocks) (% of flocks) (% of intestinal lymph nodes)

Austria 26.0 5.4 25.5 2.1

Belgium 29.2 12.4 17.8 13.0
Bulgaria 15.2 – 0.0 19.9
Cyprus 16.9 9.1 57.6 13.1
Czech Republic 78.1 19.3 42.7 5.8
Denmark 29.1 1.6 4.0 8.0
Estonia 28.4 2.0 – 6.4
Finland 45.3 0.1 0.0 0.0
France 11.1 6.2 13.3 18.5
Germany 30.4 15.0 9.2 12.7
Greece 2.6 24.0 16.5 21.2
Hungary 59.4 68.2 78.5 11.6
Ireland 7.8 27.6 27.6 15.4
Italy 4.5 28.3 38.8 16.4
Latvia 39.2 6.2 – 5.4
Lithuania 58.9 2.9 5.3 1.7
Luxembourg 42.0 – – 16.0
Malta 38.7 – – –
Netherlands 13.6c 7.5 14.1 8.5
Poland 24.3 58.2 26.9 6.4
Portugal 1.9 43.5 6.3 23.7
Romania 6.0 – – –
Slovakia 91.1 5.7 22.9 7.8
Slovenia 17.7 1.6 21.1 6.3
Spain 38.4d 41.2 56.3 30.7
Sweden 38.7 0.0 0.0 1.5
United Kingdom 15.6 8.2 32.2 21.8
EU Total 21.5 23.7 30.7 13.9
Top five serovars S. Enteritidis (45.0) S. Enteritidis (33.8) S. Bredeney (16.5) S. Typhimurium (40.0)
(% of isolates) S. Typhimurium (22.4) S. Infantis (22.0) S. Hadar (12.9) S. Derby (14.6)
S. Infantis (1.8) S. Mbandaka (8.1) S. Saintpaul (10.9) S. Rissen (5.8)
S. Typhimurium S. Typhimurium (3.0) S. Derby (9.8) S. Typhimurium
4,[5],12:i:-(1.5) S. Hadar (3.7) S. Kottbus (7.5) 4,[5],12:i:-(4.9)
S. Newport (0.9) S. Enteritidis (4.9)
a
European Food Safety Authority, European Center for Disease Prevention and Control, 2012. The European Union Summary Report on Trends and Sources of Zoonoses,
Zoonotic Agents and Food-borne Outbreaks in 2010. The EFSA Journal 10(3): 2597 (442 pp). doi:10.2903/j.efsa.2012.2597.
b
Reporting systems for human cases vary considerably. Human cases related to travel abroad are included.
c
Sentinel system, calculated from estimated population coverage.
d
Calculated from estimated population coverage.
e
The European Food Safety Authority, 2007. Report of the Task Force on Zoonoses Data Collection on the Analysis of the baseline survey on the prevalence of Salmonella in
broiler flocks of Gallus gallus, Part A. The EFSA Journal 98, 1−85.
f
The European Food Safety Authority, 2008a. Report of the Task Force on Zoonoses Data Collection on the Analysis of the baseline survey on the prevalence of Salmonella in
turkey flocks, Part A. The EFSA Journal 134, 1−91.
g
The European Food Safety Authority, 2008b. Report of the Task Force on Zoonoses Data Collection on the Analysis of the baseline survey on the prevalence of Salmonella in
slaughter pigs, Part A. The EFSA Journal 135, 1−111.

Salmonellosis in Humans
Nontyphoid salmonellosis in humans is one of the most
common gastrointestinal bacterial zoonoses worldwide. Sal-
monellosis peaks in warm seasons and the annual incidences
per 100 000 population varies between regions and countries
(e.g., Australia, 2009, 43.6; Canada, 2009, 18.0; European
Union (EU), 2010, 21.5 (Table 2); New Zealand, 2010, 26.2;
USA, 2010, 17.6). The incidence is probably underestimated
by a factor of between 5 and 20, and comparisons between
countries are confused by different sensitivities of the sur-
veillance systems.
All Salmonella are considered potentially pathogenic to
humans, but worldwide the serovars S. Enteritidis and S.
Typhimurium are predominant in human disease (Table 1).
The infective dose for healthy adult persons is believed to be
approximately 100 000 cells, but it can be as low as 10 cells
depending on the susceptibility of the person, the food ve-
hicle, and the strain of Salmonella.
Typically, onset of symptoms is 12−72 h after exposure, and
the duration of the illness is 4−5 days, often followed by a
period of fatigue. Symptoms are mainly gastrointestinal, often
accompanied by fever, headache, and muscle or joint pains. The
infection is usually self-limiting and clinically indistinguishable
from other common bacterial gastrointestinal infections. Se-
quelae are observed in 1 to 2% of the patients, and the mortality
rate with many serovars is a few percent. In contrast, human
infections with the host-adapted serovars (e.g., S. Dublin and
S. Choleraesuis) often leading to septicemia, require antibiotic
treatment, and show mortalities of 20−30%.
Food vehicles, commonly associated with sporadic and
outbreak-related salmonellosis, are table eggs, and fresh pork,
beef, and poultry meats. Increasing attention is being paid to
nonanimal foods (fruit, vegetables, fresh herbs, sprouts) as
vehicles for Salmonella. Various models have been developed
to relate human cases to Salmonella reservoirs.
Mechanisms of Pathogenicity
Six serovars carry virulence (spv) genes on virulence plasmids.
These include the host specific/adapted zoonotic serovars, and
S. Typhimurium and S. Enteritidis.
Infection Cycle
Salmonella has both an animal and an extra-animal phase.
Salmonella can survive in humid environments (soil, slurry) for
months and in dry matter for years. Once ingested, Salmonella
has to survive passage through the stomach. Although the
stomach is perceived as presenting a harsh acidic environment,
recent studies suggest that a pH gradient allows substantial
fractions of ingested Salmonella to pass to the duodenum alive.
On entering the small intestine the pH increases and the
bacteria face intestinal proteases, bile salts, the gut microbiota,
and the release of inhibitory antimicrobial peptides. Further-
more, Salmonella has to pass the mucus layer and overcome
secreted antibodies (mainly IgA) before invasion of the in-
testinal epithelium provides the organisms with a relative safe
Microbiological Safety of Meat | Salmonella spp. 371
intracellular compartment. Multiplication in the epithelial
lining leads to shedding of Salmonella in feces.
Virulence Mechanism
Salmonella controls its own invasion of enterocytes and M-cells
associated with intestinal lymphoid tissue. The uptake in
M-cells occurs in close connection with macrophages, which
process and exposes Salmonella antigens directly to the im-
mune system. Salmonella is transported in macrophages to the
lymph nodes. From there Salmonella enters the lymphatic
system and the blood stream, from which it is taken up by
macrophages in liver and spleen. If Salmonella growth exceeds
its uptake by macrophages, generalized infection results. Sal-
monella can also be taken up directly from the intestinal lumen
by dendritic cells (DC). Loosening of tight junctions enables
DCs in the submucosa to protrude through the intercellular
space and take up Salmonella directly from the lumen.
Epidemiology
Reservoirs
The primary reservoir for nontyphoid Salmonella is the gas-
trointestinal tract of animals and humans. Fecal counts of
106 cfu g−1 and 1012 cfu g−1 are common in the first week of
subclinical and clinical infections, respectively. Asymptomatic,
intermittent shedding may continue for months. Salmonella
may also be isolated from intestinal lymph nodes, the oral
cavity, and tonsils of infected animals and from blood culture
of septicemic individuals.
Birds and wild animals, including rodents, particularly in
the environments of infected farms and near dumps with
human waste, may harbor Salmonella. Manure, slurry, or
human sludge used as fertilizer and irrigation water of poor
hygienic quality can contaminate crops used for animal feed or
human consumption.
Transmission to Meat Animals
In populations of farmed animals in which Salmonella is
prevalent, exposure of meat animals to Salmonella occurs
mainly through direct or indirect fecal−oral transmission from
animals brought in from outside the herd, or from other
animals in the herd as a result of insufficient cleaning, drying,
and disinfection of pens. In poultry production there is an
additional vertical transmission from hens to chickens from
internal contamination of eggs (often by S. Enteritidis). The
serovars S. Typhimurium, S. Infantis, S. Derby, S. Dublin, and
S. Enteritidis are examples of serovars that are mainly isolated
from and transmitted by animals. Introduction of Salmonella
into herds and maintenance of Salmonella infection within
herds may also occur via infected wild birds or rodents and
contaminated equipment or personnel. When Salmonella is not
prevalent in farmed animals, the importance of transmission
of Salmonella from contaminated animal feed or feed in-
gredients increases.
372 Microbiological Safety of Meat | Salmonella spp.
Salmonella in Animal Feed
In animal feed, Salmonella is found mainly as a contaminant of
oil seeds (soy and canola) and in feed of animal origin, but the
prevalence in commercial heat-treated feed often is below 1%
of samples. The most common serovars in meat animals
are rarely contaminants of feed whereas other serovars,
often referred to as ‘exotic’ or ‘feed-borne’ serovars (e.g., S.
Livingstone, S. Mbandaka, S. Putten, S. Rissen, and S. Senf-
tenberg), are rare in meat animals and humans but common
in feed.
Because sampling of the huge amounts of feed used for
meat animals is necessarily limited, monitoring of Salmonella
in feed by culture methods is inadequate for control of Sal-
monella. In many countries end product monitoring is sup-
plemented with process control based on processors’ own
check programs and hazard analysis critical control point
(HACCP), and in several countries heat treatment or acidifi-
cation of commercial feed is part of Salmonella control
programs.
Salmonella in Meat Animals
Monitoring of Salmonella in meat animals may be conducted in
herds or at slaughter, by detection of Salmonella in fecal material,
dust, or internal organs, or by detection of antibodies against
Salmonella in blood samples, egg yolk, muscle fluid, or milk.
Poultry
Salmonella is commonly isolated from broiler chickens
worldwide, although large differences between countries exist.
In a baseline study in 23 EU Member States during 2005–06,
Salmonella was detected in 23.7% of the flocks (Table 2). More
than 50% of the isolates were S. Enteritidis or S. Infantis.
Targeted mandatory surveillance programs for S. Enteritidis
and S. Typhimurium in the EU have led to reduced flock
prevalence, determined in routine monitoring to be 4.1% in
2010 (0.4% target serovars). Salmonella is also commonly
isolated from other poultry. An EU baseline study in 2006–
2007 found Salmonella in 30.7% of turkey flocks (Table 2).
Common serovars in turkeys are S. Hadar, S. Heidelberg, and
S. Saintpaul. Commercial flocks of geese and ducks frequently
harbor a wide variety of Salmonella serovars.
Pigs
With the exception of some, mainly North European countries,
Salmonella is widespread in pig populations. In an EU baseline
study in 2006–2007, 13.9% of intestinal lymph nodes from
slaughter pigs were positive for Salmonella, with S. Typhimur-
ium and S. Derby being the predominant serovars (Table 2).
Prevalence of Salmonella in slaughter pigs is as high as 40%, as
measured by various sampling techniques, have been reported.
An EU baseline study found boars and sows culture positive in
approximately 30% of breeder and production units. An in-
creasing number of countries worldwide have implemented
surveillance programs for Salmonella in pigs. The Danish in-
tegrated program, which covers the entire production con-
tinuum, is among the most comprehensive of these. The pig
herds are classified on the basis of serology and microbiology,
enabling risk-based prevention and control actions to be taken
in herds and at slaughter.
Cattle
The occurrence of Salmonella in cattle is often low (0–1%) but
variable, with reports of 5–10% of animals shedding Sal-
monella being rare. S. Dublin and S. Typhimurium are the
predominant serovars in cattle. Owing to the severe illnesses
from S. Dublin infection in humans and cattle, this serovar
deserves special attention as a target for control efforts. Sal-
monella in cattle is mainly detected as a result of animals
showing clinical symptoms. Comprehensive monitoring
programs in the primary stages of production are rare. In
Denmark, a S. Dublin eradication program was initiated in
2007. It is based on routine serological monitoring of milk or
blood samples from all cattle herds in the country. The pro-
gram has significantly reduced the prevalence of seropositive
dairy and other herds.
Salmonella at Slaughter
Salmonella in poultry slaughterhouses
In many countries, decontamination of carcasses is part of
normal slaughter procedures for broilers, for which there is a
lack of effective slaughter hygiene measures. Decontamination
has only recently been accepted in the EU. Salmonella carcass
prevalence varies considerably between countries. In a US study,
approximately 10% of carcasses were positive for Salmonella.
In the EU a declining trend has been observed for Salmonella
in broiler carcasses, parallel to the declining prevalence of
Salmonella-positive broiler flocks, after targets for reduction of
S. Enteritidis and S. Typhimurium in broiler flocks were set.
Salmonella in pig slaughterhouses
Salmonella is widespread in slaughter pigs in many countries.
During transport and lairage Salmonella will be exchanged
between pigs, which will add to the Salmonella load on the
slaughter line. Recent epidemiological studies suggest that
direct contamination between carcasses is not the main driver
of carcass contamination but rather the many stages of pro-
cessing at abattoirs, in particular the operations of carcass
polishing and splitting. There are large variations between the
extents of Salmonella contamination of carcasses at different
plants, and many factors are likely to contribute to this. Be-
tween 1% and 8% of dressed carcasses have been reported to
be Salmonella positive.
Salmonella in cattle slaughterhouses
S. Typhimurium and S. Dublin are often isolated from cattle. In an
American study during 1998−2000, Salmonella was isolated from
70% of cattle hides, 13.3% of fecal samples, and 6.7% of carcasses,
and most frequently during August to October. Seasonality in
Salmonella prevalence has been reported from Europe also.
Salmonella in Cutting Plants
Meat can be cross-contaminated during fabrication of cuts,
and too high temperatures may allow growth of Salmonella on
meat and meat plant equipment. HACCP-based own control
 Microbiological Safety of Meat | Salmonella spp. 373

systems in cutting plants should enable the industry to The prevalence of Salmonella in meat cuttings from Danish
maintain product hygiene. Dutch data suggests that if Sal- butcher shops was 8.1% compared to a prevalence in meat
monella contamination is high on meat delivered for cutting, cuttings from supermarkets of 2.6%.
only little control over contamination of product with Sal-
monella can be expected from the plants’ own HACCP-based
control systems. Control and Preventive Measures

Control and Preventive Measures on Farms

Salmonella at Retail
The purpose of control strategies for Salmonella in meat animal
The microbiological status of retailed meat is largely deter-
farming is to reduce or eliminate Salmonella in animals that are
mined by the hygienic adequacy of handling practices and
presented for slaughter. The need for preharvest control de-
temperature control during distribution. At retail, the mincing
pends on subsequent post harvest control. Focus of preharvest
process in particular adds to cross contamination and possibly
control varies with the animal species, but the basic principles
allows growth of Salmonella. Minced beef and pork both pre-
of preharvest control are common for all. These are:
sent risks to consumers from Salmonella. In Ireland in 2007, a
Salmonella prevalence of 5% has been reported as compared to – Prevent introduction into the herd
1.8% in Denmark in 2006, whereas a prevalence up to 50% – Prevent transmission within the herd
was estimated for minced meats in The Netherlands (1998). – Reduce prevalence and shedding in infected populations.

Table 3 Main risk factors and recommendations for reduction of subclinical Salmonella in slaughter pig herds

Risk Protective factors/Recommendations

Purchase of animals Purchase of infected pigsa Purchase from noninfected supplier herd
High number of supplier herdsb
Management Continuous production Strict batch production (all-in/all-out)b,c
Mixing of pigs One-way flow of pigs – no mixingd
Slurry flooding Keep low levels in slurry pitsc
Hygiene Insufficient cleaning of pens Thorough cleaning and desiccation and proper choice and use of disinfectantc
Transmission from tools/boots Separate tools and boots for each unitc
Transmission from rodents/herd Rodent control and other biosecurityb,c
environment
Feed and feeding Dry feedd,e,f Wet feed – preferably fermented to pHo4.5
Alternatively acidification of feed (0.5–1% formic- or lactic acidg or wheyb)
Finely grinded feed cornh Coarser grinding of feed corn
Commercial pelleted feedb,d,f,h Home-mixed meal feed
Alternatively 25% of corn fed as nonheat treated, coarsely grindedi
Low % of barley in feed cornj 425% barley of feed corn
Herd size Increasing herd sizea,d,e but less Practical and financial potential to implement the reduction means above
Salmonella in the largest herdsd
a
Kranker, S., Dahl, J., Wingstrand, A., 2001. Bacteriological and serological examination and risk factor analysis of Salmonella occurrence in sow herds, including risk factors
for high Salmonella seroprevalence in receiver finishing herds. Berliener und Münchener Tierarztliche Wochenschrift 114, 350−352.
b
Lo Fo Wong, D.M.A., Dahl, J., Stege, H., et al., 2004. Herd-level risk factors for subclinical infection in European finishing-pig herds. Preventive Veterinary Medicine 62,
253−266.
c
Alban, L., Baptista, F.M., Møgelmose, V., et al., 2011. Salmonella surveillance and control for finisher pigs and pork in Denmark − A case study. Food Research International
46, 656665. Available at: http://dx.doi.org/10.1016/j.foodres.2011.02.050 (accessed 08.11.11).
d
Dahl, J., 1997. Cross sectional epidemiological analysis of the relations between different herd factors and Salmonella seropositivity. Epidemiologie et Sante Animale, 31−32,
1−3.
e
van der Wolf, P.J., Wolbers, W.B., Elbers, A.R.W., et al., 2001. Herd level husbandry factors associated with the serological Salmonella prevalence in finishing pig herds in
The Netherlands. Veterinary Microbiology 78, 205−219.
f
Bager, F., 1994. Salmonella in Danish pig herds. Risk factors and sources of infection. In: Proceedings from the XVII Nordic Veterinary Congress, pp. 79–82. Reykjavik,
Iceland: The Icelandic Veterinary Association.
g
Dahl, J., Wingstrand, A., Baggesen, D.L., Nielsen, B., Thomsen, L.K., 1996. The effect of a commercial organic acid preparation on seroprevalence and shedding of Salmonella
in finishing pigs. In: Proceedings of the International Symposium on the Epidemiology and Control of Salmonella, p. 178. Italy: Bologna.
h
Jørgensen, L., Dahl, J. Wingstrand, A. 1999. The effect of feeding pellets, meal and heat treatment on the salmonella-prevalence in finishing pigs. In: Monetti, P.G., Vignola,
G. (Eds.), Proceedings of the 3rd International Symposium on the Epidemiology and Control of Salmonella in Pork, pp. 308−312. Washington, DC: USA.
i
Dahl, J., Jørgensen, L., Wingstrand, A., 1999. An intervention study of the effect of introducing Salmonella controlling feed strategies in Salmonella high prevalence herds.
In: van der Wolf, P.J. (Ed.), Proceedings of the 4th International Symposium on the Epidemiology and Control of Salmonella and Other Food Borne Pathogens in Pork,
pp. 340−342. Germany: Leipzig.
j
Jørgensen, L., Kjærsgaard, H.D., Wachmann, H., Jensen, B.B., Knudsen, K.E.B., 2001. Effect of wheat bran and wheat:barley ratio in pelleted feed on Salmonella prevalence
and productivity of finishers. In: Proceedings of the 4th International Symposium on the Epidemiology and Control of Salmonella and Other Food Borne Pathogens in Pork,
pp. 112−114. Germany: Leipzig.
374 Microbiological Safety of Meat | Salmonella spp.
Control in poultry farms
In poultry production the very efficient vertical dissemination
of Salmonella through the breeding system demands a
top−down eradication control strategy. Successful elimination
of Salmonella from broiler flocks has taken place in many
countries. Good farming practices, with strict batch production
and biosecurity in confined production systems, and heat
treatment of commercial poultry feed have been important
elements. Thus, the flock prevalence in Danish broiler flocks
was reduced from approximately 70% in 1990 to few percent
since 2001. Reductions of Salmonella in other types of poultry
have been achieved with similar procedures.
Control in pig farms
Eradication of Salmonella from pig farms has taken place in
only a few North European countries with an a priori low
prevalence, but monitoring and reduction of Salmonella in pig
populations in which it is endemic have become more com-
mon. Risk factors for high prevalence of Salmonella in pig herds
are well known, and it has proven possible to reduce high
infection levels to moderate or low levels by purchase of
noninfected pigs, strict all-in/all-out production, improved
hygiene, and biosecurity. The use of certain feed and feeding
practices can cause microbiological and chemical changes that
reduce proliferation of Salmonella in the gastrointestinal tract
(Table 3).
Control in cattle farms
In the last decade, evidence-based interventions to control S.
Dublin in cattle herds have been developed. Introduction of
Salmonella from purchased animals must be avoided, and
introduction from equipment and herd environments may be
prevented by strict hygiene and biosecurity measures. In in-
fected herds, the risk of triggering an outbreak of salmonellosis
can be reduced by good hygiene in calving and calf units.
Infection from cow to calf through contact after calving should
be avoided by supplementing calves with colostrum from a
colostrum bank, and a one-way flow of animals in the farm
should be established.
Vaccination against Salmonella is mainly used with poultry
and may under some circumstances and for some serovars
reduce Salmonella in the flocks. Serological vaccine reactions in
routine monitoring assays should be taken into account when
vaccination is considered.
Antibiotics should never be used to control subclinical
infections with Salmonella due to the risk of development of
antimicrobial-resistant Salmonella.
Control and Preventive Measures Postharvest
Salmonella cannot be dealt with through classical meat in-
spection practices as production animals mostly are asymp-
tomatic carriers. In many countries physical (e.g., steam) and
chemical (e.g., lactic acid or chlorine) decontamination of
carcasses is used to reduce pathogen levels. Laboratory and in-
line investigations of decontamination effects show that hot
water and steam on average reduce Salmonella levels by 100-
to 1000-fold, whereas the effect of chemical decontamination
often is within the range of 10- to 30-fold. The use of two or
more decontamination methods is common and can further
improve efficacy. In Denmark, serological classification of
herds has been used to direct 1% of all pig carcasses to
hot-water decontamination. Plant control systems are often in
place with routine monitoring of slaughter hygiene. This can
also be required in relation to export to countries such as the
USA. In the USA, Salmonella performance standards at dif-
ferent levels in the meat chain have been mandated, and in
the EU process hygiene criteria have been instituted.
See also: Animal Breeding and Genetics: Traditional Animal
Breeding. Conversion of Muscle to Meat: Slaughter-Line Operation
and Pig Meat Quality. Cutting and Boning: Traditional. Foodborne
Zoonoses. Hazard Analysis Critical Control Point and Self-
Regulation. Manure/Waste Management: Manure Management.
Microbial Contamination: Decontamination of Fresh Meat;
Microbial Contamination of Fresh Meat; Microbial Contamination of
Processed Meat. Microbiological Analysis: DNA Methods; Standard
Methods. Modeling in Meat Science: Microbiology; Refrigeration.
Nutrition of Meat Animals: Pigs. Risk Analysis and Quantitative
Risk Management. Slaughter-Line Operation: Cattle; Pigs;
Poultry. Species of Meat Animals: Cattle; Pigs; Poultry
Further Reading
Alban, L., Baptista, F.M., Møgelmose, V., et al., 2011. Salmonella surveillance and
control for finisher pigs and pork in Denmark − A case study. Food Research
International 46, 656–665. Available at: http://dx.doi.org/10.1016/j.
foodres.2011.02.050 (accessed 08.11.11).
Department of Agriculture (1996). Pathogen reduction: Hazard analysis and critical
control points (HACCP) systems; Final Rule. Federal Register. Department of
Agriculture, Food Safety and Inspection Service. USDA.
Anonymous, 2005. Commission regulation (EC) No 2073/2005 of November
2005 on microbiological criteria for food stuffs. European Commission. Official
Journal of the European Union.
Bager, F. 1994. Salmonella in Danish pig herds. Risk factors and sources of
infection. In: Proceedings from the XVII Nordic Veterinary Congress, pp. 79−82.
Reykjavik, Iceland: The Icelandic Veterinary Association.
Berends, B.R., Knapen, F.V., Mossel, D.A.A., Burt, S.A., Snijders, J.M.A., 1998.
Impact on human health of Salmonella spp. On pork in the Netherlands and the
anticipated effects of some currently proposed control strategies. International
Journal of Food Microbiology 44, 219–229.
Borch, E., Nesbakken, T., Christensen, H., 1996. Hazard identification in swine
slaughter with respect to food borne bacteria. International Journal of Food
Microbiology 30, 9–25.
Dahl, J., 1997. Cross sectional epidemiological analysis of the relations between
different herd factors and Salmonella seropositivity. Epidemiologie et Sante
Animale 31−32, 1–3.
Dahl, J., Jørgensen, L., Wingstrand, A., 1999. An intervention study of the effect of
introducing Salmonella controlling feed strategies in Salmonella high prevalence
herds. In: van der Wolf, P.J. (Ed.), Proceedings of the 4th International
Symposium on the Epidemiology and Control of Salmonella and Other Food
Borne Pathogens in Pork, pp. 340−342. Germany: Leipzig.
Dahl, J., Wingstrand, A., Baggesen, D.L., Nielsen, B., Thomsen, L.K., 1996. The
effect of a commercial organic acid preparation on seroprevalence and shedding
of Salmonella in finishing pigs. In: Monetti, P.G., Vignola, G. (Eds.), Proceedings
of the International Symposium on the Epidemiology and Control of Salmonella,
p. 178. Italy: Bologna.
European Food Safety Authority, European Centre for Disease Prevention and
Control, 2012. The European summary report on trends and sources of
zoonoses, zoonotic agents and food-borne outbreaks in 2010. EFSA Journal 10,
442.

Grimont, P.A.D., Weill, F.-X., 2007. Antigenic formulae of the Salmonella serovars,
9th edition France: WHO Collaborating Centre for Reference and Research on
Salmonella. Institut Pasteur.
Hald, T., Vose, D., Wegener, H.C., Koupeev, T., 2004. A Bayesian approach to
quantify the contribution of animal-food sources to human salmonellosis. Risk
Analysis 24, 255–269.
Hald, T., Wingstrand, A., Swanenburg, M., Altrock, A., Thorberg, B.-M., 2003. The
occurrence and epidemiology of Salmonella in European pig slaughterhouses.
Epidemiology and Infection 131, 1187–1203.
Hansen-Wester, I., Hensel, M., 2001. Salmonella pathogenicity islands encoding
type III secretion systems. Microbes and Infection 3, 549–559.
Jørgensen, L., Dahl, J., Wingstrand, A. 1999. The effect of feeding pellets, meal and
heat treatment on the salmonella-prevalence in finishing pigs. In: Bahnson, P.B.
(Ed.), Proceedings of the 3rd International Symposium on the Epidemiology and
Control of Salmonella in Pork, pp. 308−312. Washington, DC, USA: Biomedical
Communications Center, College of Veterinary Medicine, University of Illinois at
Urbana-Champaign.
Jørgensen, L., Kjærsgaard, H.D., Wachmann, H., Jensen, B.B., Knudsen, K.E.B.,
2001. Effect of wheat bran and wheat: barley ratio in pelleted feed on Salmonella
prevalence and productivity of finishers. In: Proceedings of the 4th International
Symposium on the Epidemiology and Control of Salmonella and Other Food
Borne Pathogens in Pork, pp. 112−114. Germany: Leipzig.
Kranker, S., Dahl, J., Wingstrand, A., 2001. Bacteriological and serological
examination and risk factor analysis of Salmonella occurrence in sow herds,
including risk factors for high Salmonella seroprevalence in receiver finishing
herds. Berliener und Münchener Tierarztliche Wochenschrift 114, 350–352.
Kühler, H., Sakaguchi, T., Hurley, B.P., et al., 2007. Salmonella enterica serovar
Typhimurium regulates intercellular junction proteins and facilitates transepithelial
neutrophil and bacterial passage. American Journal of Physiology:
Gastrointestinal & Liver Physiology 293, G178–G187.
Lo Fo Wong, D.M.A., Dahl, J., Stege, H., et al., 2004. Herd-level risk factors for
subclinical infection in European finishing-pig herds. Preventive Veterinary
Medicine 62, 253–266.
O’Connor, A.B., Denagamage, T., Sargeant, J.M., Rajic, A., McKean, J., 2008.
Feeding management practices and feed characteristics associated with
Salmonella prevalence in live and slaughtered market-weight finisher swine: A
systematic review and summation of evidence from 1950 to 2005. Preventive
Veterinary Medicine 87, 213–228.
Smid, J.H., Heres, L., Havelaar, A.H., Pielaat, A., 2012. A biotracing model of
Salmonella in the pork production chain. Journal of Food Protection 75,
270–280.
The European Food Safety Authority, 2007. Report of the Task Force on Zoonoses
Data Collection on the Analysis of the baseline survey on the prevalence of
Microbiological Safety of Meat | Salmonella spp. 375
Salmonella in broiler flocks of Gallus gallus, Part A. The EFSA Journal 98,
1−85.
The European Food Safety Authority, 2008a. Report of the Task Force on Zoonoses
Data Collection on the Analysis of the baseline survey on the prevalence of
Salmonella in turkey flocks, Part A. The EFSA Journal 134, 1−91.
The European Food Safety Authority, 2008b. Report of the Task Force on Zoonoses
Data Collection on the Analysis of the baseline survey on the prevalence of
Salmonella in slaughter pigs, Part A. The EFSA Journal 135, 1−111.
The European Food Safety Authority, 2009. Analysis of the baseline survey
on the prevalence of Salmonella in holdings with breeding pigs, in the
EU, 2008, Part A: Salmonella prevalence estimates. EFSA Journal 7(12), 1377
(93 pp).
Threllfall, E.J., Wain, J., Lane, C., 2011. Salmonellosis. In: Palmer, S., Soulsby, L.,
Torgerson, P., Brown, D.W.G. (Eds.), Zoonoses. Biology, clinical practice and
public health control, 2nd ed. New York: Oxford University Press Inc., pp. 992.
van der Wolf, P.J., Wolbers, W.B., Elbers, A.R.W., et al., 2001. Herd level husbandry
factors associated with the serological Salmonella prevalence in finishing pig
herds in The Netherlands. Veterinary Microbiology 78, 205–219.
World Health Organization, Global Foodborne Infections Network. Country Databank.
Available at: http://thor.dfvf.dk/portal/page?_pageid=53,1&_dad=portal&_
schema=PORTAL (accessed 30.03.12).
Relevant Websites
http://www.cdc.gov/
Centers for Disease Control and Prevention (CDC) USA.
http://ecdc.europa.eu/en/Pages/home.aspx
European Centre for Disease Prevention and Control (ECDC), Sweden.
http://www.efsa.europa.eu/
European Food Safety Authority (EFSA), Italy.
http://www.ozfoodnet.gov.au/
OzFoodNet, Australia.
http://www.nml-lnm.gc.ca/NESP-PNSME/index-eng.htm
Public Health Agency of Canada, National Enteric Surveillance Program.
http://www.surv.esr.cri.nz/index.php?we_objectID=2933
Public Health Surveillance, Information for New Zealand Public Health Action.
http://thor.dfvf.dk/portal/page?_pageid=53,1&_dad=portal&_schema=PORTAL
World Health Organization, Global Foodborne Infections Network, Country
Databank.