Table of content:

Introduction……………………………………………2-3
Taxonomy………………………………………………4
Description……………………………………………..5-6
Scientific classification……………………………….8
Epidemiology …………………………………………8-9
Pathogenesis ………………………………………….10-11
Small noncoding RNA………………………………..11
Nomenclature ……………………………………….12
Diagnosis……………………………………………13
Treatment……………………………………………14
References…………………………………………..16-19
Introduction
Salmonella Enteritidis is a pathogen that infects both
animals and humans, causing gastroenteritis or systemic infections.
To colonize the host, S. enterica must overcome the acidity of the
stomach, the microbial flora of the gut, and the intestinal barrier in
the host (ÁLVAREZ-ORDÓÑEZ et al., 2012). This study evaluated S.
enterica serovar Enteritidis under various stress conditions, as it is
the most prevalent serovar among the brazilian human population
(ROWLANDS et al., 2014). Although the main source of Salmonella
are eggs and egg products (GANTOIS et al., 2009), other foods such
as raw milk, beef and pork was associated to S. enterica infection
outbreaks (ROWLANDS et al., 2014; CDC, 2015). Several reports
documented the presence of S. Enteritidis in swine abattoirs in Brazil
(BESSA et al., 2004; KICH et al., 2011). Strategies used in abattoirs to
prevent S. enterica growth include lower storage temperatures and
surface decontamination using acids (SUN et al., 2003; PIPEK et al.,
2006). Although these strategies create environments of moderate
stress, S. enterica is capable of adapting to these adverse conditions,
not only in natural environments in animal or human hosts, but also
in commercial ones such as abattoirs and food industries (WINFIELD
& GROISMAN, 2003; ÁLVAREZORDÓÑEZ et al., 2012).

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 However, stress factors encountered during processing and
storage of food as well as during passage through host physiological
barriers (For e.g. gastric fluid) had a significant effect on the survival
of S. enterica. The physiological pH of human fluid gastric can range
from 1.5 to 3.5 (MARIEB & HOEHN, 2010). Studies reported that S.
enterica is resistant to acid and low temperatures conditions
(MÜLLER et al., 2012). Bacteria adapted to mild acid stress survived
in similar or different stress conditions due to a cross protection effect
(SPECTOR & KENYON, 2012). Moderate acidic conditions could
trigger resistance to gastric fluid in S. enterica, increasing the risk and
severity of illness (YUK & SCHNEIDER, 2006).

There was significant increase in the prevalence of S. Enteritidis

and incidence of human salmonellosis in recent years. Moreover, acid
tolerance is an important virulence factor related to survival of
foodborne pathogens at low pH conditions of the gastric barrier.
Studying the stress factors, therefore contributes to the resistance
development in S. Enteritidis, which became increasingly relevant.
The objective of this study was to evaluate the effect of pH and low
storage temperatures on S. Enteritidis and its subsequent growth in
simulated gastric fluid (SGF).

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Taxonomy

The genus Salmonella is part of the family of Enterobacteriaceae. Its

taxonomy has been revised and has the potential to confuse. The
genus comprises two species, S. bongori and S. enterica, the latter of
which is divided into six subspecies: S. e. enterica, S. e. salamae, S. e.
arizonae, S. e. diarizonae, S. e. houtenae, and S. e. indica.The
taxonomic group contains more than 2500 serotypes (also serovars)
defined on the basis of the somatic O (lipopolysaccharide) and
flagellar H antigens (the Kauffman–White classification). The full
name of a serotype is given as, for example, Salmonella enterica
subsp. enterica serotype Typhimurium, but can be abbreviated to
Salmonella Typhimurium (Brenner FW. et al., 2000).

Further differentiation of strains to assist clinical and

epidemiological investigation may be achieved by antibiotic sensitivity
testing and by other molecular biology techniques such as pulsed-field
gel electrophoresis, multilocus sequence typing, and, increasingly,
whole genome sequencing. Historically, salmonellae have been
clinically categorized as invasive (typhoidal) or noninvasive
(nontyphoidal salmonellae) based on host preference and disease
manifestations in humans (Brenner FW. et al., 2000).

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Description
Salmonella is a Gram-negative, intracellular pathogens ,facultative
anaerobic bacteria, belonging to the family of Enterobacteriaceae
(Yan SS. et el., 2004 ). Salmonella spp. is widely distributed in the
environment, but the intestinal tract of animals is the main habitat of
the bacteria (Le Minor 2003). Salmonella contamination occurs
through the consumption of contaminated foods like egg, milk and
poultry meat (Gillespie B. et al., 2003). Twenty percent of world
poultry products are contaminated with Salmonella, and they can
persist for a long time in the animal and human environments and
facilities through biofilm formation (Vestby LK. et al., 2009). In most
of the salmonellosis outbreaks resulted from poultry products
consumption, Enteritidis and Typhimurium serovars have been
isolated (Vose D. et al., 2011). In Iran, poultries are reported to be the
predominant reservoirs for Salmonella enterica and serovar
Enteritidis was isolated in 51.4% (35/68) of the samples (Zahraei-
Salehi T. et al., 2005). S. enterica, serovar Enteritidis is implicated in
60% of salmonellosis in European people and is the world’s leading
cause of salmonellosis (Thorns C. 2000). In the United States, S.
Typhimurium is mostly associated with Salmonellosis (CDC
1999).Multidrug-resistant (MDR) due to Salmonella is known as a
major public health problem around the world and there is an
increased use of antibiotics in human and animal settings (Hsu SC. et

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al., 2006). MDR Salmonella may be transmitted to human throughout
the production chain and so important risk factors have been
identified during processing (Heyndrickx M. et al., 2002).

Polymerase Chain Reaction (PCR) is a valuable method for

investigating food-borne outbreaks and pathogens identification
(Riyaz-Ul-Hassan S. et al., 2004). PCR provides fast results and a high
degree of specificity. The incorporation of a routine PCR test in
combination with traditional culture methods could be effective in
providing a more accurate profile of the prevalence of this pathogen in
broiler carcasses (Carrasco E. et al., 2012).

Because the main source of carcasses contamination with Salmonella

are intestinal tract, skin and feathers of chickens, which may develop
along the processing line (Whyte P. et al., 2002), this study was
conducted on carcasses of a slaughterhouse to evaluate the
contamination rate of poultry carcasses with Salmonella spp.,
especially S. Enteritidis and S. Typhimurium in a poultry processing
plant in Mashhad, Iran, using culture and multiplex PCR method by
detecting prot6E and fliC genes(YUK & SCHNEIDER, 2006).

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6
Scientific classification

Domain: Bacteria

Phylum: Proteobacteria

Class: Gammaproteobacteria

Order: Enterobacterales

Family: Enterobacteriaceae

Genus: Salmonella

Species: S. enteritidis

Epidemiology

Most cases of salmonellosis are caused by food infected with S.

enterica, which often infects cattle and poultry, though other animals
such as domestic catsand hamsters have also been shown to be sources
of infection in humans. Investigations of vacuum cleaner bags have
shown that households can act as a reservoir of the bacterium; this is
more likely if the household has contact with an infection source (i.e.,
members working with cattle or in a veterinary clinic) ( Swanson SJ. et
al., 2007).

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Raw chicken eggs and goose eggs can harbor S. enteritidis, initially in
the egg whites, although most eggs are not infected. As the egg ages at
room temperature, the yolk membrane begins to break down and S.
enterica can spread into the yolk. Refrigeration and freezing do not
kill all the bacteria, but substantially slow or halt their growth.
Pasteurizing and food irradiation are used to kill Salmonella for
commercially produced foodstuffs containing raw eggs such as ice
cream. Foods prepared in the home from raw eggs, such as
mayonnaise, cakes, and cookies, can spread salmonellae if not
properly cooked before consumption(ÁLVAREZ-ORDÓÑEZ et al.,
2012).

S. enteritidis genomes have been reconstructed from up 6,500 year old

human remains across Western Eurasia, which provides evidence for
geographic widespread infections with systemic S. enterica during
prehistory, and a possible role of the Neolithization process in the
evolution of host adaptation. Additional reconstructed genomes from
colonial Mexico suggest S. enterica as the cause of cocoliztli, an
epidemic in 16th-century New Spain(MARIEB & HOEHN, 2010).

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Pathogenesis

Salmonella species are facultative intracellular pathogens.Salmonella

can invade different cell types, including epithelial cells, M cells,
macrophages, and dendritic cells.As facultative anaerobic organism,
Salmonella uses oxygen to make ATP in aerobic environment (i.e.,
when oxygen is available). However, in anaerobic environment (i.e.,
when oxygen is not available) Salmonella produces ATP by
fermentation; by substituting one or more of four less efficient
electron acceptors than oxygen at the end of the electron transport
chain: sulfate, nitrate, sulfur, or fumarate (LaRock et al., 2015).

Most infections are due to ingestion of food contaminated by animal

feces, or by human feces, such as by a food-service worker at a
commercial eatery. Salmonella serotypes can be divided into two main
groups—typhoidal and nontyphoidal. Nontyphoidal serotypes are
more common, and usually cause self-limiting gastrointestinal disease.
They can infect a range of animals, and are zoonotic, meaning they
can be transferred between humans and other animals. Typhoidal
serotypes include Salmonella Typhi and Salmonella Paratyphi A,
which are adapted to humans and do not occur in other animals
(LaRock et al., 2015).

Secreted proteins are of major importance for the pathogenesis of

infectious diseases caused by S. enteritidis. A remarkably large

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number of fimbrial and nonfimbrial adhesins are present in
Salmonella, and mediate biofilm formation and contact to host cells.
Secreted proteins are also involved in host-cell invasion and
intracellular proliferation, two hallmarks of Salmonella pathogenesis
(Hensel M 2009).

Small noncoding RNA

Small nonprotein-coding RNAs (sRNA) are able to perform specific

functions without being translated into proteins; 97 bacterial sRNAs
from Salmonella Typhi were discovered.

AsdA (antisense RNA of dnaA) is a cis-encoded antisense RNA of

dnaA described in S. enterica serovar Typhi. It was discovered by deep
sequencing and its transcription was confirmed by Northern blot and
RACE analysis. AsdA is estimated to be about 540 nucleotides long,
and represents the complementary strand to that encoding DnaA, a
protein that plays a central role in the initiation of DNA replication
and hence cellular division (GANTOIS et al., 2009).

In rich media, it is highly expressed only after reaching the stationary

growth phase, but under limiting iron or osmotic stress, it is already
expressed during exponential growth. Overexpression of AsdA
stabilizes dnaA mRNA, increasing its levels and thereby enhancing its
rate of translation. This suggests that AsdA is a regulator of DNA
replication (GANTOIS et al., 2009).

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Nomenclature

S. enterica has six subspecies, and each subspecies has associated

serovars that differ by antigenic specificity. S. enterica has over 2500
serovars (Vestby LK. et al., 2009).

Salmonella bongori was previously considered a subspecies of S.

enterica, but it is now the other species in the genus Salmonella. Most
of the human pathogenic Salmonella serovars belong to the enterica
subspecies. These serogroups include S. Typhi, S. Enteritidis, S.
Paratyphi, S. Typhimurium, and S. Choleraesuis. The serovars can be
designated as written in the previous sentence (capitalized and
nonitalicized following the genus), or as follows: "S. enterica subsp.
enterica, serovar Typhi"(Vestby LK. et al., 2009).

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Diagnosis

Isolation and identification. In clinical cases direct plating on

Brilliant Green and McConkey agar may be adequate. Enrichment
media such as buffered peptone followed by selective broth or semi-
solid media (e.g. Rappaport-Vassiliadis) followed by plating on two
selective media will greatly increase sensitivity. However this has the
potential to reveal the presence of salmonellae that are irrelevant to
the clinical problem under investigation(BESSA et al., 2004).

Differentiate from Pullorum/Typhoid, other enterobacteria such as E.

coli. S.Enteritidis causes cross-reactions which may be detected with
S.Pullorum serum agglutination tests. It is possible to detect reactions
with specific antigens in agglutination tests but competitive and direct
Elisa tests are more commonly used today ( KICH et al., 2011).

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Treatment
Medications:
Because salmonella infection can be dehydrating, treatment focuses
on replacing fluids and electrolytes. Severe cases may require
hospitalization and fluids delivered directly into a vein (intravenous).
In addition, your doctor may recommend:

 Anti-diarrheals. Medications such as loperamide (Imodium A-D)

can help relieve cramping, but they may also prolong the
diarrhea associated with salmonella infection (YUK &
SCHNEIDER, 2006).
 Antibiotics. If your doctor suspects that salmonella bacteria have
entered your bloodstream, or if you have a severe case or a
compromised immune system, he or she may prescribe
antibiotics to kill the bacteria. Antibiotics are not of benefit in
uncomplicated cases. In fact, antibiotics may prolong the period
in which you carry the bacteria and can infect others, and they
can increase your risk of relapse. Sulphonamides, neomycin,
tetracyclines, amoxycillin, fluoroquinolones in accordance with
the sensitivity. Good management. Chemotherapy can prolong
carrier status in some circumstances(YUK & SCHNEIDER,
2006).

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14
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