Salmonella, Shigella and Yersinia

● These are primary pathogens that are part of the enterobacteria family

Salmonella
● Its a coliform bacilli
● They are motile gram negative facultative anaerobes
● They are non-lactose fermenyters
● Glucose: Acid (+) and Gas (+) (this is the main difference between salmonella and shigella)
● They are resistant to bile salts
● They produce H2S
● They are catalase positive

Classification
● DNA homology classifies salmonella into 2 species, salmonella enterica is the main one and its further
divided into 6 subspecies, and the other species is Salmonella bongori
● There are thousands of serotypes in those 2 species and most can cause disease in humans
● Salmonella enterica serotype Enteritidis and Salmonella enterica serotype Typhimurium, the two most
important serotypes of Salmonella transmitted from animals to humans

Antigenic composition of Salmonella

● They have O antigen, H antigen, and Vi antigen
● Vi antigen is found in S.typhi and S.hirschfeldi
○ Its involved in the pathogenicity of the bacteria
○ It inhibits the agglutination of O
antigens with the O antibodies
● O antigen is capsular antigen and H is
flagellar antigen

Virulence factors
● It has an endotoxin that helps in intracellular
survival
● S typhi and some strains of S paratyphi
have a capsule
● It has both fimbrial and non fimbrial
adhesion
● Enterotoxins: salmonella strains may
produce heat labile enterotoxins, they cause
diarrhea
● Outer membran proteins: involved in the
ability of salmonella to survive inside macrophages
● Acid tolerance response gene (ATR): protects salmonella from stomach acid and the acidity of
phagosomes
● Catalase and superoxide dismutase: they protect the bacteria from intracellular killing
● Flagella: they provide motility
● Pathogenicity islands: there are 2 PAIs on the bacterial chromosome that regulate attachment,
engulfment and replication (very important)
○ PAI 1 contains gene that code for salmonella-secreted invasion proteins (Ssps) and – a
type III secretion system that injects protein into the host cell
○ PAI 2 contains genes that allow the bacteria to escape the immune response of the host,
and has a 2nd type 3 secretion system
Pathogenicity
● After the bacteria is ingested and passes through the stomach, salmonella attaches to the mucosa of the
small intestine
● They invade M cells in peyer patches as well as invade neterocytes
● The bacteria stay in endocytic vacuoles where they replicate and kill the hocy cell and spread to
adjacent cells
● Our inflammatory response keeps the bacteria in the gastrointestinal tract, we release proastaglandins,
this stimulates the secretion of cAMP, and this causes diarrhea (fluid loss)
● Bacteria can also be transported to the blood or lymph

Diseases caused by Salmonella

● Check the pic
● It can cause bloody diarrhoea
● 1- Gastroenteritis: (food poisoning)
○ Its the most common form of salmonellosis (the name
of infections caused by salmonella)
○ Happens after eating contaminated food or water
○ S.enteritidis is the most common strain source
○ The bacteria doesnt enter the blood. Its found in
stool instead
How does food poison happen?
-salmonealla is absorbed into the epithelium of the small intestine
-bacteria penetrates cells and migrates to the lamina propria
-they multiply in lymph nodes and then move to the reticular system
-they cause hypertrophy and hyperplasia
-this stimulates an inflammatory response where prostaglandins are released. This stimulates cAMP and fluid
loss, leads to diarrhea
○ It may be caused by any species but Salmonella Typhi, • Salmonella Paratyphi, and • Salmonella
Choleraesuis are the most common
○ The bacteria is found in blood, but not found in stool or intestines
○ The risk of developing bactermia is high in kids and old people, and people wth a weak immune
system
● 3- Typhoid Fever (Enteric fever)
○ Salmonella typhi produce typhoid fever
○ S. paratyphi A, Salmonella Schottmuelleri
(formerly Salmonella Paratyphi B), and
Salmonella Hirschfeldii (formerly Salmonella
Paratyphi C) produce paratyphoid fever which
is milder than typhoid
○ Typhoid fever occurs when the bacteria leaves
the intestine and multiplies within cells of the
reticuloendothelial cells
○ The bacteria then renters the intestine and
cause food poisoning symptoms

Asymptomatic colonization
● The species of salmonella that cause typhoid and paratyphoid fever can stay in the body after causing the
disease
● They stay without causing symptoms
● The gallbladder is the reservoir for this
Diagnosis
● If we have enteric/typhoid fever we collect specimens from blood, urine, stool or bone marrow
● Food poisoning: stool, vomit, suspected contaminated food
● Speticemia: blood
● We carry out biochemical tests to identify the species
● We carry out serotyping to determine whether this species is dangerous/significant
● We carry out the widal test for serotyping
● Cultures:
○ To test for the presence of salmonella in the blood stream we use blood culture
○ To test for the presence of salmonella in stool, we use specific media like Selinit F broth, EMB,
Mackonkey and Endo agar
● We use multiplex PCR

NAAT
● They are multiplex DNA amplification tests that are used for GI infections
● The biggest advantage is that this test can identify many common enteric bacteria, viruses and parasites
at the same time

Epidemiology
● Most infections happen after eating contaminated food like chicken, eggs and dairy products. In children
they can be transmitted from fecal contaminated food
● S. typhi and S.paratyphi are strict human pathogens and don't infect animals. Theyre passed person to
person. Asymptomatic long term colonization occurs commonly
● Infections happen mainly in the warm months of the year

Treatment
● Gastroenteritis/food poisoning: antibiotics are not given becuase they will prolong the inflammation and
infection
● Typhoid fever: antibiotics are given to avoid asymptomatic colonization: Ciprofloxacin, chloramphenicol,
SXT can be used
● There are 2 vaccines that reduce the risk of disease fo travellers going in areas with many infecte
dindivdiuals: an oral live attenuated vaccine and a Vi capsular polysachharide vaccine
—---------------------------------------------------------------------------------------------------------------------------------------------------
Shigella
● They are also primary pathogens from the enterobacteria family
● There are 4 species of shigella.
○ S. sonnei is responsible for most infections in developed countries
○ S. flexneri: responsible for infections in developing countries as well but not as common
○ S. dysenteria: responsible for the most severe infections
○ S. boydi is not commonly found
● They are all mannitol positive except S. dysentria

General Features
● They are non motile
● Most strains cannot ferment lactose (s. Sonnei can slowly ferment it)
● Glucose: Acid (+) and Gas (-)
○ This means that the bacteria can ferment glucose and produces acid as a byproduct, but does not
produce gas during fermentation
● They are resistant to many drugs
● They are resistant to bile salts
● There are 4 species like mentioned above, theyre classified based on their O antigen
Pathogenicity and Virulence Factors
● Shigella is resistant to the acid of the stomach and can pass normally through it and reach the intestines
● In the intestine it invades and destroys the mucosa of the colon

Virulence factors:
● An exotoxin (Shiga toxin)
● It invades cells and can survive inside the cells and replicate
● They have large plasmids which ocntain lpa genes necessary for virulence. These genes help in the
attachment and entry process of the bacteria
● They have a type 3 secretion system that secretes 4 lpa proteins (lpaA, lpaB, lpaC, lpaD) into the
epithelial cells and macrophages
● These proteins induce membrane ruffling (changes in the cell membrane), which allows the bacteria to be
engulfed and enter inside the cell

Continuation of Pathogenicity: Invasiveness

● The bacteria pass through the stomach and small intestine and invades the colon
● Shigella attaches and invades M cells first. M cells usually transport foreign bacteria from the intestines to
the macrophages beneath. So shigella invades M cells and goes inside the macrophages
● Inside the macrophages shigella escapes the phagosome and goes into the cytoplasm where it activates
apoptosis (Cell death) in the macrophage. This differs from salmonella as salmonella multiplies in
the phagocytic vacuole
● After they exit the macrophage they go into host cells. Within the host cell, actin filaments propel the
bacteria through the cytoplasm into adjacent epithelial cells, so shigella can move from cell to cell,
thereby avoiding the antibody mediated humoral immunity similar to Listeria Monocytogenes
○ They migrate from cell to cell through disrupted tight junctions
● Shigella survives phagocytosis into the macrophage by inducing programmed cell death
● The process leads to the release of Il-1 beta, which results in the attraction of PNLs (leukocytes) into the
infected tissue
● This destabilizes the integrity of the intestinal wall
and allows the bacteria to reach deeper epithelial
cells
This is explained further:
● Bacteria released from the dead macrophages
contact the enterocytes(host cells) and start a
multistep invasion process mediated by the invasion
plasmid antigens mentioned earlier (lpaA-lpaD)
● As the bacteria moves from cell to cell, this destroys
the enterocytes and creates ulcers in the colon
mucosa
● These ulcers allow shigella to enter the lamina propria
(deeper than the epithelium) and start an intense
inflammatory response
● In healthy people the infection doesnt spread beyond the
lamina propria (doesnt go to the bloodstream)
● The diarrhea created by this process is due to the
inflammation. Its made up of small-volume stools
(pebbles) that contain WBCs, RBCs, bacteria. “This is
classic dysentery” So shigella leads to bloody diarrhea
Shigella Flexneri and Samonella typhi: differences and
similarities
● Shigella and salmonella start by invading M cells
● Shigella multiplies in The M cell and propels through the
cytoplasm to invade adjacent cells
 ● Salmonella passes through the M cell and goes to the submucosa where its taken up by macrophages.
● Salmonella is able to multiply in macrophages in the lymph node and other reticuloendothelial cells
● Both organisms induce apoptosis in their host cells, In shigella this leads to a mucosal ulcer and in
salmonella this leads to spreading to the bloodstream and causing typhoid fever

Shiga Toxin
● S. dysenteria strains produce an exotoxin called shiga toxin
● Shigatoxin is enterotoxic, neurotoxic and cytotoxic
● Its coded by chromosomal structures
● Its an A-5b dimer toxic
● Enterotoxic effects:
○ It blocks the absorption of electrolytes, glucose, amino acids from the intestinal lumen
○ This is the opposite of cholera toxin (by vibrio cholera) and the labile toxin of enterotoxigenic e coli
(ETEC) which act by blocking absorption of Sodium but hypersectrion of water and Cl-, K+,
HCO3- ions out of the enterocytes and into the lumen
● Cytotoxic effects
○ The B subunit of shiga toxin binds to glycolipids on the surface of th ehost cell
○ The A subunit enters the cell through receptor mediated
endocytosis
○ The A subunit cleaves the 28 sRNA in the 60s Ribosome, which
prevents the binding of aminoacyl transfer RNA. This causes:
■ Inhibition of protein synthesis
■ Cell death
■ Hemorrhage/blood loss (blood and fecal leukocytes in
stool)
■ In a few individuals, shiga toxin can damage golmerular
endothelial cells and result in renal failure
● Neurotoxic Effects
○ Fever and abdominal pain/cramping
—---------------
Clinical Diseases and Symptoms
● Shigella causes disease by invading and replicating in cells lining the colon
● The most common form of disease is gastroenteritis (food poisoning), also called as Bacillary dysentery
and shigellosis
● Shigella causes acute inflammatory colitis and bloody diarrhea
● Shigellosis is mainly a pediatric disease
● The infective dose required to cause is very low
● Shigella is characterized by abdominal cramps, diarrhea, fever and bloody stools
● The severity of the disease depends on the species present
○ S. dysenteriae is the most pathogenic followed by S. flexneri, S. sonnei and S. boydii
Bacillary Dysentery/Shigellosis
● Humans are the only reservoir
● Transmission: through the fecal oral route
● Symptoms: watery diarrhea, bloody stools with mucus and pus, fever, tenesmus
● There is sharp pain in the stomach
● Nausea and Vomiting

Complications
● Reactive arthritis
○ (previosuly known as Reiters syndrome): its a form of inflammatory arthritis that develops in
response to an infection
○ it can be seen in people with HLA-b27 antigen
○ Its a late complication of S flexneri infection
○ Its also seen in enteric pathogens such as salmonella, campylobacter and yersinia
 ● Hemolytic Uremic Syndrome (HUS): it occurs after S.dysenteria
● Comvulsions in children
● Rectal Prolapse and Rectum Perforations (DO NOT GOOGLE IT)

Epidemiology
● Humans are the only reservoir
● They can spread person to person by fecal oral route
○ This means: The bacteria is present in the feces of a contaminated person, if someone goes and
touches surfaces in the bathroom they may touch contaminated fecal particles

Immunity
● Infections with one serotype of shigella provide protection against the same type in the future, But this
protection is modest and does not extend to other serotypes
● The main defense against shigella infections is a component called SIgA (secretory immunoglobulin A)

Treatment
● Antibiotic therapy shortens the duration of the disease
● “Treatment should be guided by in vitro susceptibility tests”
● Antibiotics: ciprofloxacin, ceftriaxone, azithromycin

Diagnosis
● We take stool specimens
● We culture the specimens
● They form lactose negative colonies on EMB and Macconkey agar
● On TSI medium: they are lactose and sucrose negative. Glucose: acid positive but gas negative. They
also dont produce H2S
● We can use shigella antisera to perform a slide agglutination test
—---------------------------------------------------------------------------------------------------------------------------------------------------
Yersinia
● Yersinia is a bacterial family containing multiple species
○ Yersinia Pestis: is a highly virulent pathogen that causes the very fatal disease known as the
Black Plague
○ Yersinia enterocolitica: it causes enterocolitis and transfusion related septicemia and endotoxic
shock
○ Yersinia pseudotuberculosis: its uncommon, it causes gastroenteritis and mesenteric
lymphadenitis thats similar to acute appendicitis

General Characteristics
● Yersinia is a cocobacillary bacteria (looks round and rod shaped at the same time)
● They retain staining at both ends of the cell, this pattern is known as bipolar staining
● Yersinia are fermenters
● They are oxidase and Urease negative
● They are catalase positive
● Y.peptis is covered with a protein capsule
● Some specie slike Y.enterocolitica can grow at cold temperatures (so can still grow in the fridge)

Pathogenesis
● Enteropathogenic yersinia enter the human in contaminated food and invade M cells in peyer patches
● They have multiple virulence factors such as invasin and yersinia outer membrane proteins (Yops) that
allow it to invade host cells
● Invasin is one of the surface adhesion molecules on yersinia thats required for the initial step of invasion
of M cells. Invasins bind to integrins on the surface of host cells
 ● Yops are injected into the host cell by type 3 secrteion system. Yops trigger cytotoxic events like
disrupting biochemical reactions (dephosphorylation and serine kinase), sensor function and changing the
actin cytoskeleton. All these end up with the cell dying
● Pathogenic yersinia species is resistant to phagocytic killing, this is done thanks o the type 3 secretion
system
● Type 3 secretion system also suppresses cytokine production

Yersinia Pestis
● It has a capsule, plasminogen activator and Fibrolysin
● Its resistant to serum killing
● It has the ability to absorb organic iron as a result of siderophore independent mechanisms
● Y. pestis has 2 plasmids that code for virulence factors
○ The fraction 1 gene (f1 gene-know it as formula 1 gene) codes for the protein capsule that gives it
antiphagocytic properties
○ Plasminogen activator gene (pla gene) code for a protease enzyme that breaks down C3b and
C5a
○ Both these genes prevent opsonization and phagocytosis

Clinical Forms of the plague (Black Death)

● The first form was the buponic plague, which had swollen and painfull axillary and inguinal lymph
nodes
○ It was transmitted from mammals
by flea (arthropod) bites or
contact with contaminated animal
tissue
● The 2nd form was pneumonic plaque,
which was spread person to person
through the respiratory route

Epidemiology
● Yersinia are mainly animal pathogens, where they can spread to humans. Humans are accidental hosts
● There are 2 forms of Y.pestis infection:
○ Urban plague and Sylvatic plague
● Pigs, rodents, livestock and rabbits are the natural reservoirs for Y. enterocolitica
● Rodents, wild animals and game birds are the natural reservoirs for Y. pseudotuberculosis

There are cycles that describe how the plague spreads in different environment from wild animals to humans
-The first cycle is the Sylvatic cycle: fleas on infected rodents such as mice and prairie dogs pass the infection to
other animal species and also can pass it to us and caus ethe buponic plague
-The 2nd cycle is the urban plague cycle: masses of rats are closer to humans, bites from infected fleas transmit
it to many humans and other rats, the infection spreads a lot more. In both the urban and sylvatic cycle, buponic
plague develops in the human
- if Y pestis spreads into the blood it can react the lungs and cause pneumonic plague. Pneumonic plague can
be spread between humans through the respiratory route

Arthropod-Borne transmission of Plague

● Fleas are needed for the spread of the plague
● Fleas ingest the bacteria when theyre feasting on the blood of dead infected animals
● Fleas have coagulase enzyme which forms a fibrin clot in the throat of the flea, this causes a blockage
and imits how much food passes. So the flea always remains hungry and eats more
● Fleas vomit the infectious material into new host

Epidemiology of the plague continued

● Its a zoonotic infection (infection for animals) and humans are accidental hosts
Diagnosis
● Y pestis is a gram negative coccobacillus that has bipolar staining with giemsa, wrights or waysons
staining
● Immunofluorescent staining is rapid
● It grows aerobically on most culture media including
blood agar and maconkey agar

Treatment
● Y.pestis infections are treated with streptomycin like
tetracyclines, chloramphenicol or
trimethoprim-sulfamethoxazole