Salmonella

Salmonella One

infect all types of domestic animals

of the most prevalent agents causing foodborne disease-salmonellosis

Classification

is based on serology and phage susceptibility assays

More S.

than 2600 serovars

enterica is divided into seven subspecies with group I causing disease in humans and other warmblooded animals
Examples

:Typhimurium, Enteritidis, Choleraesuis, Dublin, Gallinarum, and Pullorum

Salmonella
Salmonella Facultative Salmonella Resistant

are gram (-) motile rods aerobes grow at an optimum of 37oC

to heat if in foods with lower pH, or low water activity or in food with a high fat content
Viability

declines during frozen storage, especially if near the freezing point

Optimum

pH for growth of Salmonella is 6.5-7.5; may grow at a pH range of 4.5-9.0

Succinic

and formic acid reduce the persistence of Salmonella in foods

Salmonella Prevalence

Factors which favor the continued presence in the food chain

Ubiquity of salmonella in the environment Intensive husbandry practices in the meat, fish, and shellfish industries Lack of microbiological control on animal feeds favors the continued prevalence of this pathogen in the food chain

Poultry products remain the main reservoir of salmonella Eggs and egg products are also a concern since there is Transovarian transmission of the pathogen into the interior of the egg

Salmonella Prevalence

Developed countries= more than 80% of the Salmonella cases occur individually rather than in outbreaks Large Salmonella Outbreaks

Foods linked to transmission include milk powder, raw milk, cheddar cheese, egg products, and liver pate Some fruits and vegetables have also been linked to outbreakscantaloupes, chocolate, mustard dressing

Infectious dose will vary, as few as 1 to 10 cells

Foods with a high fat content may have a low infectious dose due to the organism being trapped in micelles which protect it against acidic ph

Salmonella Prevalence

Food-borne infections account for 1.3 billion cases of acute diarrhea with 3 million deaths world-wide Salmonellosis in the U.S. is 40,000 cases annually Recent years a notable increase in cases related to a multi-drug resistant S. typhimurium DT104 Case-fatality and hospitalization rates due to this strain are twice that of other Salmonella infections

Salmonella Pathogenesis
Disease
Disease

depends on age of the host-more severe in newborns, infants, the elderly

Disease
oHost

also depends on the serovar and type of host

specific serovars: typhi, paratyphi, sendai cause disease only in humans

oPullorum/gallinarum

in poultry, dublin in cattle; choleraesuis serovars in pigs can also infect humans; typhimurium and enteritidis are the major serovars that cause disease in humans, cattle, poultry, sheep, pigs, horses, and wild rodents.

Salmonellosis

Salmonellosis
Diarrhea,

abdominal pain, vomiting and fever

Bacteremia

has been associated with highly invasive serovars such as choleraesuis or dublin
Other

clinical sequels include erythrema nodosum, meningitis, osteomielitis, septic arthritis, pnuemonia, choleocydtitis, endocarditis, pericarditis, and cystitis
A

complication of infection is the establishment of a carrier state which lasts for 5-6 months.
Carrier

state is responsible for the spread of the bacterium to others, farms, food-handlers, and consumers.

Salmonella Invasion
Salmonella

passes through the stomach and survives the acid pH by eliciting an acid tolerance response
Colonizes

the distal ileum; portal of entry is the M-cell and it binds by means of fimbriae from the lpf operon or from the pcf operon
Heritable

phase variation mechanism

Triggered

by intestinal-derived

signal

Salmonella Invasion

Penetration and destruction of M-cell

Localized accumulation of cytoskeletal proteins such as actin, vinculin, vimentin, and ezrin at site of entry
Formation of membrane ruffles occurs with every cell type examined

Recruitment of lymphoid cells and PMN

Secretion of large amount of fluid Leads to inflammation of the area

Salmonella Invasion

Colonizes intestinal epithelial cells In vivo

Microvilli denuded transiently, membrane ruffling Bacterial penetration 1-2 h Cytosketetal rearrangements, 30-60 min Actin filament rearrangement No microtubule involvement Ca++ flux Distruption of tight junctions Causes depolarization of epithelial cells

In vitro

Salmonella Invasion
Pathogenecity

island of 40kb has been identified which also contains a type III secretion system
SPI

operon encoded type III secretion system

InvA-C

operon

oFunctional

filamentous appendages appear upon cell contact and require a function type III secretion system
oActivated

by a shift from acidic to alkaline pH in the absence of eukaryotic cells; also activated by environmental conditions such as high osmolarity, low oxygen tension, DNA supercoiling, and alkaline pH
oMore

than 15 proteins are inserted into the inner and outer membrane. Classes: inner and outer membrane proteins InvA, SpaP, SpaQ, SpaR, SpaS, InvG, InvH, PrgH, PrgK; an energizer of the system, the inner membrane ATPase InvC; secreted proteins with putative targets in the host cell known as effector proteins SipB, SipC, SptP, AvrA; other secreted proteins SipA; cytosolic chaperons which prevent premature degradation of secreted proteins SicA, SicP; transcriptional regulatory proteins InvF, HilA, and InvE

Salmonella Invasion
Other

Invasion Factors
is needed LPS is also required for invasion are also needed

Motility Intact

Fimbriae

Host

Factors for Invasions

of host cell essential functions
ruffling
oActin

Subversion

cytoskeleton reorganization

oMembrane oSignal

transduciton pathways

oSopE

(substances of the SpI-1 encoded system) stimultes actin reorganization by increasing GDP/GTP nucleotide exchange in several GTPases
oStimulates

production of proinflammatory cytokines as IL-8, and activates transcriptional factors

Proliferation within Nonprofessional Phagocytes

Salmonella Salmonella

proliferates in membrane-bound vacuoles

containing vacuole fuses with host compartments containing lysosomal-membrane glycoproteins and bypasses compartments of the endocytic route
SPI-2

is required for intracellular proliferation within epithelial cells

Recent

findings suggests that macrophages and specialized epithelial cells allow Salmonella to proliferate intracellularly

Proliferation within Nonprofessional Phagocytes

Although non-growing, remain viable for periods of 1014 days Salmonella have been shown to reduce their metabolic rate to perpetuate with the host cell The PhoPQ system is involved in preventing growth of intracellualr Salmonella; phoP mutants grow actively in non-permissive cells First example of a bacterial-mediated mechanism that prevents its own proliferation in a specific host cell Long persistance of Salmonella within non-phagocytic cells could be linked to Salmonella derived reactive arthirits, chronic infections, or carrier asymptomatic

Proliferation within Professional Phagocytes

Active

membrane ruffling and macropinocytosis occurs upon ingestion of Salmonella by phagocytic ells
Trafficking

route used by Salmonella within a cultured macrophage is different than within epithelial cells
Fusion

of the endosome with mature lysosomes seems to occur

Nevertheless

that compartment does not behave as a conventional mature lysosomal since there is no processing of lysosomal enzymes contained within
Acidification

triggers bacterial response for survival and replication within these cells
Salmonella

may exist within two populations; one activity growing and the other static but not viable

Proliferation within Professional Phagocytes

Salmonella

is also able to trigger apoptosis of the infected

macrophage
Products

involved in triggering apoptosis are: SPI-1 oxygen regulated protein (OrgA), OmpR/EnvZ, SPI-1 secreted proteins such as InvJ, SpaO SipB, SipC, and SipD
Induced

apoptosis may destroy phagocytic cells before they process antigen and activate T-cells may react to infection with Salmonella and program its own death to avoid excessive release of immune mediators
Macrophage

Survival within Phagocytic cells

Proteins involved in DNA repair are needed such as RecA and RecBC for survival Increased levels of SodA (Mn dependent) results in increase survival within macrophages in vitro but not for survival in the mouse model Increased levels ot SodC (Zn2+-Cu2+) results in increased survival in the mouse model but not in macrophages in vitro SlyA is a transcriptional regulator involved in Salmonella survival by giving it resistance to toxic oxidative products Sap proteins are required for antimicrobial peptide resistance

Salmonellosis
PhoP-PhoQ Two

regulatory System

component system=pair of proteins, one of which, the sensor, underoges a change in response to a change in the environment and communicates this change, usually in the form of a phosphate to another protien, response regulator, which then causes the appropriate cellular response
Sensing

extracellular signals such as limiting amounts of Mg2+ and mild acidic pH

Salmonellosis

PhoQ is a inner membrane integral protein capable of binding Mg2+ and Ca2+

Upon binding PhoQ auto-phosphorylates and transfers the phosphate to PhoP, a cytosolic protein
Phosphorylated PhoP acts then as a transcriptional regulator or at least 40 genes
o

Some virulence genes are activated as pag and others repressed such as prg Virulence traits linked to this regulatory system include:(1) bacterial survival within macrophages, epithelial cell (2) resistance to cationic antimicrobial peptides, (3) invasion of epithelial cells, (4) control of Ag presentation by bacteriainfected macrophage, (5) resistance to bile acids

Salmonellosis
PmrAB
Two

regulatory system
component system

Controls

bacterial resistance to polymixin and other LPS-binding antimicrobial peptides

Auto-regulated

Expression

of these genes is dependent on low pH or Mg2+ limitation

Salmonellosis

OmpR/EnvZ regulatory system

Two component system Senses changes in external medium osmolarity OmpR is the transcriptional regulator EnvZ is its cognate membrane sensor
o

Controls expression of major outer membrane proteins:OmpC and OmpF

Salmonellosis
o

High osmolarity favors expression of OmpC while low osmolarity shifts the expression to PmpF OmpF mutants are avirulent in the mouse typhoid model OmpCF mutants are attenuated in the in vivo model when administered orally but not interperiotoneally

Salmonellosis
RpoS

(katF) Sigma factor

Important High

regulatory factor in starved bacteria

levels of this protein in cells that are reaching stationary phase

Needed Needed

for survival in stationary phase

also for nutrient stresses in Salmonella such as carbon, nitrogen, or phosphate starvations
Increased
Required

in intracellular bacteria
for virulence

Salmonellosis
Roles

of a Salmonella virulence plasmid (60-80 kb)

Needed

for the production of systemic disease; cured strains are unable to produce systemic disease in the mouse typhoid system
Mutants

retain capacity for colonization the intestinal tract and spreading to target tissues and organs
Spv

operon
control of the RpoS sigma factor

Under

Protein

play a role in signaling between intracellular Salmonella and the host cell
Only

synthesized in intracellular bacteria