Properties of the genus Salmonella

○ A member of the family Enterobacteriaceae

○ Colon flora; sometimes carried asymptomatically
○ Gram negative rods
○ Facultatively anaerobic
○ Most are motile with peritrichous flagella
○ Chemoorganotroph, metabolize by both respiratory
and fermentative pathway
○ Oxidase negative
○ Catalase negative
○ Catabolize carbohydrates; produce acid and gas
○ Generally produce H2S
Official nomenclature by CDC
 Genus Salmonella
 Two species:
1. Salmonella enterica: 2423 serovars
six subspecies – I: S. enterica supsp. enterica
II: S. enterica supsp. salamae
IIIa: S. enterica subsp arizonae
IIIb: S. enterica subsp diarizonae
IV: S. enterica subsp houtenae
VI: S. enterica subsp indica
2. Salmonella bongori (V): 20 serovars
Human pathogens
 Salmonella enterica serovar Typhi
 Salmonella enterica serovar Enteritidis
 Salmonella enterica serovar Paratyphi
 Salmonella enterica serovar Typhimurium
 Salmonella enterica serovar Choleraesuis
Diseases caused are mainly
○ Enteric or Typhoid fever
○ Uncomplicated enterocolitis
○ Systemic disease
 Enteric Fever
 Typhoid: Salmonella typhi
 Paratyphoid: Salmonella paratyphi A, B, C
 These two strains are well adapted for invasion and survival
in host tissues
Symptoms
Incubation period: ranges from 7-28 days
After about 10-14 days, the disease begins with
 lethargy
Malaise
fever
generalized pain
Constipation
2nd week (bacteremia)
○ Prolonged and spiking fever
○ Sensitive stomach, Diarrhea
○ Abdominal pain
○ Headache
○ Prostration
Enterocolitis
 Cause: nontyphoid Salmonella spp.
 Examples
○ Salmonella enteriditis
○ Salmonella gallinarum
○ Salmonella typhimurium
 Appears 8-72 hours after contact with the pathogen
 Symptoms:
○ Cramps
○ Nausea
○ Vomiting
○ Diarrhea
 Self-limiting
 Remission within 5days of onset of symptoms
 treatment is by water & salts replacement
 Antibiotics are not usually needed
 in infants, elderly or immunosuppressed may be
complicated by bacteremia
 Lead to infection in the meninges, joints, bone
 mortality rate of 10-20%
 Use of antibiotics might prolong the carrier state and
intermittent excretion
Systemic infection (Septicemia)
caused by - Salmonella typhimurium
- Salmonella paratyphi A and B
- Salmonella choleraesuis
Mainly affect young children, adults and immunocrompromised
Other infections:
Various chronic conditions like - Reactive arthritis
- Ankylosing spondylitis
 Especially in those with preexisting physiological,
anatomical and immunological disorders
 Distribution
 The primary habitat is the intestinal tract of animals such as
birds, reptiles, farm animals, humans, and occasionally
insects
 Animal fecal matter
 Animal hides
 maintained within an animal population by
 Non symptomatic animal infections
 in animal feeds
 slaughter animals reinfected in a cyclical manner
Secondary contamination:
 handlers
 direct contact of non contaminated foods with
contaminated foods
 TRANSMISSION
• Infection follows ingestion of contaminated food or water.
•Meat, poultry, eggs & diary products are frequent sources.

• Pets, domestic animals and infected human are potential

reservoirs
•Person to person & animal to human transmission is
recognized
• In healthy humans a dose of about one million bacteria is
necessary to produce symptoms.
The Salmonella Food-Poisoning
Syndrome
 Symptoms usually develop in 12–14 hours, although
shorter and longer times have been reported
 The symptoms consist of
 Nausea
 Vomiting
 abdominal pain (not as severe as with staphylococcal
food poisoning),
 Headache
 chills,
 and diarrhea
 usually accompanied by prostration
 Muscular weakness
 Faintness
 moderate fever
 restlessness, and drowsiness.
 Symptoms usually persist for 2–3 days
 The average mortality rate is 4.1%
 varying from 5.8% during the first year of life, to
2% between
 the first and 50th year, and 15% in persons over
50 years
 Among the different species of Salmonella,S.
Choleraesuis has been reported to produce the
highest mortality rate—21%
 up to 5% of patients may become carriers of the
organisms
 Numbers of cells on the order of 107–109/g are
generally necessary for salmonellosis
Pathogenesis
Bacteria in oral cavity

Pass to intestinal tissues and mesenteric lymph follicles

phagocytes migrate to the infected tissue

increased mucus secretion and mucosal inflammation releasing

prostaglandins

Adenylate cyclase in intestinal epithelial cells activated

Increased fluid secretion into the lumen (Diarrhoea)

failure of host to control Salmonella

lead to septicemia and other chronical clinical conditions

 Attachment
 Bacterium attach and invade intestinal columnar epithelial cells
(enterocytes) and M cells in Peyer’s patches
 Depends on-
 successful competition with normal flora for attachement sites
 Evasion of sIgA present on epithelical cells
 Colonization involves interaction between host glycoprotein
receptors on the intestinal surface with
 bacterial Type I or type 3 fimbriae
 Surface adhesins
 Nonfimbriate hemagglutinins
 Enterocyte induced polypeptides
 Motility is not essential for adherence
 Increases frequency of contacts between pathogen and
host
 Some Proteinaceous invasion appendages develop upon
contact with epithelial cells-
 Thicker than flagella
 thinner than type 1 fimbriae
 Assembly is energy-dependent
 Short lived
 shed with the appearance of membrane ruffles on
epithelial cells
 Invasion
Different ways:
1. uptake by antigen-sampling M cells
2. phagocytic uptake by macrophages present in the
epithelial monolayer
3. Self mediated entry into phagocytic and non-phagocytic
cells using a type III secretion system 1 (T3SS1)
4. attachment and internalization via a T3SS1-independent
way
Phagocytic uptake
 Non-specific
 Involves interaction between Pattern-recognition receptors
on phagocytes and pathogen-associated molecular
patterns:
○ LPS
○ Flagellin
T3SS1-mediated invasion
 highly specific process
 depends on the tightly regulated expression of a number of
bacterial factors
 Following adherence, Salmonella promote Ca2+ influx to
host cell
 It acts as a signal that influences actin polymersiation
 A subset of SPI-1 effector proteins are delivered via T3SS
 function to induce membrane deformation and actin
rearrangement (‘membrane ruffling’)
 SipA and SipC: induce actin polymerization
 SopE2: activates signal transduction cascade involved in
cytockeletal rearrangement
 Bacterial uptake by pinocytosis
 enclosed within an intracellular phagosomal compartment
Salmonella-containing vacuole (SCV)
 maturing SCV moves towards the Golgi apparatus
 positioned within the perinuclear area
 the SCV-enclosed bacteria replicate
 characterized by formation of Salmonella-induced filaments
(Sifs, SifA)
 SCV move towards the basal pole of host cell
 Released into laminar propria
 remain localised to the intestine
 Typhoid: - survives in intestinal macrophages
- disseminate to the liver and spleen via the
bloodstream and lymphatic system
 Following engulfment, cytoskeleton returns back to its
resting state
 mediated by SptP
○ Disrupts actin cytoskeleton
○ Reverses ion of actin filaments and membrane
ruffles into their initial state
Evasion of host defense
1. Enzymes to neutralize toxic oxygen radicals; eg.,
Superoxide dismutase
 About 30 proteins are synthesized
2. The phoP/phoQ two component transcriptional regulator
system
 Enables survival within high acidity of phagolysosomes
 Encodes cell surface protease to degrade defensin
 Outer membrane protein PagC (serum resistance)
3. SPI-2 effectors
 SpiC: inhibits endosome-endosome fusion and
phagosome-lysosome fusion
4. SPI-3 : needed for intracellular survival
Eg., products of mgtCB
Inflammation
 SPI-1 effectors induce acute intestinal inflammation
1. Stimulate epithelial cells to produce cytokines (eg., IL-8)
 polymorphonuclear leukocytes (PMNs) are recruited
 SopE,SopE2,SopB
2. Induce release of PMN attractant
 SipA
3. induce apoptosis of epithelial cells, neutrophils and
macrophages
 intensifies inflammation
 SipB, Flagellin
Diarrhea

Caused by several ways

1. SopB, SopE, SopE2 and SipA:
 Inflammation and PMN transmigration
 Disruption of intestinal epithelial cell tight junctions

2. SopB:
promotes cellular chloride ion secretion and fluid flux
Virulence factors
 Fimbriae and non-fimbrial adhesins
 T3SS -1
 T3SS-2
 Type I secretion system
 Flagella
 ion transporters
Fimbriae
 Salmonella have 13 predicted fimbrial loci
 many are induced in vivo
 required for
 biofilm formation
 attachment to host cells
 colonization
 not required for intracellular survival
T3SS1
 Encoded by Salmonella pathogenicity island-1 (SPI-1)
 Contigiuos and functionally related loci located within a
40-50kb segment of chromosomal DNA
 Deliver at least 15 effectors involved in invasion and
post-invasion processes
 Many have multiple activities within host cells
T3SS2
 Encoded by SPI-2
 Function of effectors are not clear
 required for
○ survival within macrophages
○ systemic virulence in the mouse
several are involved in
 SCV positioning and intracellular replication : SseF and
SseG
 the formation of Sifs: SifA
Flagella and flagellin
 Flagellar-based motility can increase the invasiveness of
Salmonella
 Controversial

Type I secretion system

 Secretes two surface-associated proteins BapA and SiiE
 Involved in invasion/adhesion
Virulence plasmid (SPV)
 Present in Salmonella serovars Typhilurium
- Dublin
- Gallinarum
- Enteritidis
- Cholerasuis
- Pullorum
- Abortusovis
 Absent in S. typhi
 8kb in size
 Involved in systemic spread and infection of extra intestinal
tissues
 Not in adhesion and invasion of epithelial and M cells
 Enables rapid multiplication in host cell
 Evasion of host defense
 Lysis of macrophage
 Elicit inflammatory response
 Induce enteritis in host

Signals that trigger spv transcription are-

 Hostile environment within macrophage
 Iron limitation
 Elevated temperature
 Low pH
 Nutrient deprivation
 Contains Five genes: spvR
SpvR: transcriptional regulator of spvABCD
 Function remains unclear
 both SpvB and SpvC are translocated via the
T3SS2
SpvB:
 ADP-ribosyl transferase
 associated with host cell cytotoxicity
SpvC:
 phosphothreonine lyase
 Act on host signal transduction
 The plasmid contains a fimbrial operon pef
 Encodes adhesin
 Colonization of small intestine
Superoxide dismutase
 Salmonella uses a superoxide dismutase, SodCI
 confers protection from extracellular reactive oxygen
species
 SodCI is ‘tethered’ within the periplasm
 protease resistance
 allows this enzyme to function in the harsh environment of
the phagosome
Iron acquisition
 in response to iron deprivation Salmonella produce two
siderophores:
 Enterochelin or enterobactin
 salmochelin (a glucosylated derivative of enterobactin)
 this modification may be important for resistance to
lipocalin-2, an antimicrobial protein that prevents
bacterial iron acquisition in the inflamed intestinal
epithelium
 Synthesis of siderophores regulated by the fur (ferric
uptake regulator) gene
 Degree of virulence related to siderophore content
 Another mechanism identified within SPI-I
Mechanism of iron uptake
 Siderophore binds Fe+3
 Fe+3 - Siderophore complex formed
 Complex binds outer membrane receptor (induced by
low intracellular iron concentration)
 Complex transported into cytoplasm
 Fe+3 reduced to Fe+2 state
 Release of Fe+2 (low affinity of siderophore to Fe+2 )
Mg2+ acquisition
 Salmonella has three distinct systems for uptake
of Mg2+:
○ CorA
○ MgtA
○ MgtB
 each is essential for virulence
MgtC:
 encoded on the same operon as MgtB
 not a Mg2+ transporter
 required for intra macrophage survival
 and growth in magnesium-depleted medium
Zn and K uptake system
 K+ and Zn2+ are required for intracellular survival

The ZnuABC uptake system:

 high-affinity Zn uptake system
 required for growth of S. typhimurium in low-zinc
conditions and intracellularly in some cultured cells
 ZnuABC mutants are defective for virulence in both
susceptible and resistant mouse strains
The Trk system:
 a multiunit protein complex
 functions as a low-affinity K+ transporter
 may function in resistance to antimicrobial peptides
Enterotoxin
 A virulence phenotype in Salmonella serovars, including S
typhi
 Encoded by a chromosomal gene stx
 activates adenylate cyclase (located in epithelial cell
membrane)
 Increase in the cytoplasmic concentration of cAMP in host
 Fluid movement out of cell into lumen
 Increased Cl-secretion and reduced Na+ absorption
 role in intracellular invasion and the subsequent
pathogenesis is unclear
 Production of other cytotoxic proteins has also been
reported in nontyphoid salmonellae
Cytotoxin
 Some strains produce a thermolabile cytotoxin
 Localized in the outer membrane
 Inhibits protein synthesis
 Lyses host cells
 Thus promotes spread of bacteria into host tissues
 Addition of Ca2+ blocks the toxin in vitro
 Indicates that it may acts a chelator
 This might also be an invasion protein that causes
apoptosis
Capsular polysachharide
 Capsular polysaccharide prevents opsonization of
complement
 Prevents phagocytosis
 Two genes are involved in capsule formation:
 viaA and viaB
 viaB encodes 6 proteins
 Key to the capsule formation

LPS
 Prevents lytic attack by complement system
Susceptibility and immunity
 all people equally susceptible to infection
 acquired immunity can keep longer, reinfection are rare
 immunity is not associated with antibody level of “H”,
“O”and “VI”.
 No cross immunity between typhoid and paratyphoid.