Enterobacteriaceae Classification Based on Lactose Fermentation

General Characteristics ● Several selective and differential media

● Gram negative bacilli used to isolate distinguishes between LF &
● Facultative anaerobes LNF
● All species ferment glucose with the ● The most important media are:
production of acid or acid and gas 1. MacConkey agar
● All are motile @35C with peritrichous 2. Eosin Methylene Blue (EMB) agar
flagella (except for Klebsiella , Shigel la and 3. Salmonella Shigella (SS) agar
Yersinia 4. Triple Sugar Iron (TSI) agar
● Catalase positive (except for Shigella Lactose Fermenter
dysenteriae type 1 which is catalase 1. Escherichia coli
negative) 2. Klebsiella
● Oxidase negative (except for Plesiomonas 3. Enterobacter
● Non encapsulated (except for Klebsiella and 4. Serratia (except S. fonticola ) SLOW or
Enterobacter) LATE
● Non spore forming 5. Citrobacter SLOW or LATE
● Reduce nitrate to nitrites (except for *Coliform bacilli Escherichia, Klebsiella,
Photorhabdus and Xenorhabdus) Enterobacter and Citrobacter Lactose
● Most are commensal of the GI tract Non Lactose Fermenter
(except for Plesiomonas , Salmonella , 1. Proteus
Shigella and Yersinia 2. Morganella
● Serratia and Yersinia may grow at 1C to 3. Providencia
5C 4. Hafnia
● They do not produce cytochrome 5. Edwardsiella
oxidase except for Plesiomonas 6. Salmonella
7. Shigella (except S. sonnei ––“Late” LF)
Microscopic and Colony Morphology 8. Yersinia
Gram stain Classification Based on the Clinical Infection
● Gram negative coccobacilli or straight rods 1. Opportunistic pathogens
SBA or CAP (Chocolate Agar Plate) no value for ● The opportunistic pathogens are often a part
initial identification of the usual intestinal microbiota of both
● Except for Klebsiella and sometimes humans and animals
Enterobacter large and very mucoid 2. Primary pathogens
colonies ● Plesiomonas , Salmonella , Shigella and
● Some isolates of E. coli Beta hemolytic Yersinia
● Not present as commensal biota in the GI
Differential and Selective Media tract of humans
● EMB (Eosin Methylene Blue) Virulence and Antigenic Factors
● MAC ● O antigen (somatic antigen) this is a heat
● HEA ( Hektoen Enteric Agar), XLD (Xylose stable antigen located on the cell wall.
Lysine Desoxycholate ) highly selective ● H antigen (flagellar antigen) this is a heat
- (H2S) Hydrogen Sulfide Production labile antigen found on the surface of
can be seen flagella, structures responsible for motility.
- Contains sodium thiosulfate and ● K antigen (capsular antigen) this this is a
ferric ammonium citrate produces heat labile polysaccharide found only in
blackening of (H2S)producing certain encapsulated species
colonies - K1 antigen E. coli
- Vi antigen Salmonella enterica
subsp. enterica serotype Typhi.
 IMViC Reactions
Media and Tests used for the Identification of 1. Indole Test
Enterobacteriaceae 2. Methyl Red Test
3. Voges-Proskauer Test
Triple Sugar Iron Agar (TSI) 4. Citrate Utilization Test
Indole Test
● Ability to ferment glucose, lactose, and ● Test the ability of the bacteria to produce
sucrose and to produce hydrogen sulfide the enzyme tryptophanase and deaminate
● Composition: 1% lactose, 1% sucrose, tryptophan to indole, pyruvic acid, and
0.1% glucose ammonia
● Ferrous sulfate and sodium thiosulfate ● Two reagents can be used:
are added to detect the production of 1. Ehrlich’s reagent more sensitive
hydrogen sulfide H2S (black precipitate in 2. Kovac’s reagent
the medium) ● Both contains PDAB
● pH indicator: Phenol red (Paradimethylaminobenzaldehyde)
● Original color of medium: red ● Positive reaction pink/red
● Kligler’s iron agar (KIA) is a similar ● Tryptophan
medium, but only incorporates the Methyl Red Test (MRVP MEDIUM)
carbohydrates glucose and lactose. ● If glucose is metabolized by the mixed acid
- H2S production can also be detected fermentation pathway, stable acid end
with this formulation products are produced, which results in a
low pH
Reactions on TSI agar or KIA ● Negative reaction remain yellow after
1. No fermentation Alkaline slant/alkaline butt addition of MR pH indicator
(ALK/ALK or K/K) or alkaline slant/no ● Positive reaction red color after addition of
change (ALK/no change or K/NC) Red Red MR pH indicator
2. Glucose fermentation only, no lactose (or
sucrose in TSI) fermentation Alkaline
slant/acid butt or K/A Red Yellow
3. Lactose (or sucrose or both)
fermentation Acid/acid, Yellow Yellow Voges Proskauer Test
4. H2S production blackening of the medium ● It measure the production of acetoin after
5. Gas production (aerogenic) or no gas the addition of α naphthol (catalyst or color
production ( nonaerogenic ) formation of intensifier) followed by 40% KOH or NaOH
bubbles or splitting of the medium from the ● Acetoin then oxidized into diacetyl 2,3
bottom of the tube Butanediol
● Positive reaction red complex

Citrate Utilization Test

● It determines whether an organism can use
sodium citrate as a sole carbon source
● Medium used Simmons’ Citrate
● Medium (SCA) routinely used
● Ammonium salts are the nitrogen source in
the medium and utilization of these salts
results in the release of ammonia, causing a
pH change
● pH indicator bromthymol blue
● Positive reaction blue
Nitrate Reduction Test Motility Test
● Determines whether an organism has the ● 0.4% agar
ability to reduce nitrate to nitrite ● Single stab in the medium
● Detected by the addition of N,N Dimethyl α ● Routine medium used SIM (Sulfide Indole
naphthylamine and sulfanilic acid Motility) Agar
● Positive reaction red color (diazo red dye) ● Positive reaction movement away the stab
line or hazy appearance throughout the
medium
Amino Acid Utilization
Oxidase Test ● Decarboxylase and Dihydrolase Test
● Determines the presence of the cytochrome ● Deamine Test --PAD Test (Phenylalanine
oxidase system that oxidizes reduced Deaminase Test)
cytochrome with molecular oxygen Decarboxylase and Dihydrolase Test
● Differentiates Enterobacteriaceae ● Decarboxylase tests determine whether the
(oxidase negative) from Pseudomonads bacterial species possess enzymes capable
(oxidase positive) of decarboxylating (removing the carboxyl
● Reagent tetramethyl -p-phenylenediamine group, COOH) specific amino acids in the
dihydrochloride test medium
● Positive reaction purple or lavender color ● Two amino acids commonly used lysine
Urease Test and ornithine
● Determines whether a microorganism can ● Products of decarboxylation amine or
hydrolyze urea (by the action of urease diamine molecules and CO2
enzyme), releasing a sufficient amount of ● Agmatine
ammonia to produce a color change by a pH
indicator
● Preferred medium Christensen’s Urea
Agar
● pH indicator phenol red
● Positive reaction bright pink color
● Media include Moeller decarboxylase base
● Ammonium carbonate:Proteus
medium, motility indole ornithine (MIO), or
Gelatin Hydrolysis Test
lysine iron agar (LIA)
● Bacteria that produce gelatinases that break
● Positive reaction purple
down gelatin into amino acids.
● Negative reaction yellow
● Liquefaction of the gelatin is a positive
Phenylalanine Deaminase Test (PAD Test)
test
● Determines whether an organism
Hydrogen Sulfide Production (H2S)
possesses the enzyme that deaminates
● Utilizes the sodium thiosulfate sulfur source
phenylalanine to phenylpyruvic acid
to form H2S, a colorless gas H2S combines
● Reagent 10% ferric chloride
with the indicator, ferrous sulfate producing
● Positive reaction green color
black color (positive reaction)
● Proteus , Morganella , and Providencia
● Media demonstrate the production of
species are phenylalanine deaminase
H2S
positive
- Sulfide indole motility agar
Lysine Iron Agar Slant
- Motility indole ornithine agar
● It is used primarily to determine whether the
- Hektoen enteric agar
bacteria decarboxylate or deaminate lysine
- Salmonella Shigella agar
● Useful for differentiating Salmonella spp.
- Triple sugar iron agar
(lysine positive) from Citrobacter spp. (lysine
- Kligler Iron agar
negative)
- Lysine iron agar.
● Also useful in differentiating Proteus ,
● FERROUS SULFIDE
Morganella , and Providencia spp. from
most other members of Enterobacteriaceae
 ● Butt slant medium Pathogenic Strains
● Contains amino acid lysine, glucose, ferric ● Uropathogenic E. coli
ammonium citrate and sodium thiosulfate ● Enterovirulent E. coli or Diarrheogenic E.
● pH indicator bromcresol purple coli
Categories:
● Enterotoxigenic E. coli (ETEC)
● Enteroinvasive E. coli (EIEC
● Enteropathogenic E. coli (EPEC)
● Enterohemorrhagic E. coli (EHEC)
● Enteroadherent
- Diffusely adherent E. coli (DAEC)
- Enteroaggregative E. coli (EAEC)
Uropathogenic E. coli (UPEC)
● Strain of E. coli that is considered as the
most common cause of UTIs in humans
Opportunistic Members of the Family ● Pili – primary virulence factor associated
Enterobacteriaceae with the ability of E. coli to cause UTIs
● Escherichia spp. ● Other factors include cytolysins and
● Hafnia spp. aerobactins
● Klebsiella spp. Enterotoxigenic E. coli
● Proteus spp. 10^6 to 10^10 organisms
● Edwardsiella sp. ● Associated with diarrhea of adults and
● Enterobacter spp. especially children in tropical and
● Morganella spp. subtropical climates
● Citrobacter spp. ● It is the most common cause of diarrheal
● Serratia spp. disease
● Providencia spp. ● Also referred to as “traveler’s diarrhea”
Escherichia coli (Colon Bacillus) ● High risk “Achlorydia ” deficiency of
● The most significant species in the hydrochloric acid within the stomach
genus Escherichia ● Colonization of ETEC on the small
● Associated with UTIs, diarrheal disease and intestine is mediated by fimbriae
CNS infections - Permit ETEC to bind to specific
- Most common cause of UTIs in receptors on the intestinal microvilli
humans ● Produce a heat stable toxin (ST) or a heat
● Primary marker of fecal contamination in labile toxin (LT) or both
water quality testing ● Characterized by watery diarrhea,
● Most strains are motile and generally abdominal cramps, nausea (rare), usually
possess adhesive fimbriae and sex pili no vomiting or fever
● O, H, K antigen
● Serotyping Enteropathogenic E. coli
Cultural Characteristics: ● Causes “infantile diarrhea”
● MAC lactose positive (pink) colonies - Severe diarrhea (<1 year old)
● EMB greenish metallic sheen - Outbreak occurs in hospital nurseries
and daycare centers
Escherichia coli ● Rarely seen in adults
1. Fermentation of glucose, lactose, trehalose, ● Characterized by low grade fever, malaise,
and xylose vomiting and diarrhea
2. IMViC + + - - ● Stool contains large amount of mucus, no
3. Does not produce H2S, DNase, urease, or blood present
phenylalanine deaminase
Enteroinvasive E. coli - Produces watery diarrhea, vomiting,
● Produce dysentery with direct dehydration and occasionally abdominal
penetration, invasion, and destruction of pain mostly in children
the intestinal mucosa ● WBCs and RBCs are absent from the stool
● Very similar to the diarrheal illness ● Cause of diarrhea in HIV positive patients
produced by Shigella spp. E. Coli Extraintestinal infections
● Differentiated by motility test and lactose ● One of the most common cause of
fermentation septicemia and meningitis in neonates
● May be nonmotile, NLF ● K1 capsular antigen virulence factor
● Characterized by fever, severe abdominal associated with neonatal meningeal
cramps, malaise and watery diarrhea infections
Enterohemorrhagic E. coli ● Clinically significant isolate in blood cultures
● O157: H7 strain of E. coli from adults (bacteremia)
● Associated with hemorrhagic diarrhea and ● From urogenital tract infection or from GI
HUS (hemolytic uremic syndrome) source
● HUS characterized by low platelet count, Other Escherichia species
hemolytic anemia and kidney failure E. hermannii
● Watery diarrhea --->Bloody diarrhea with ● Yellow pigmented organism; isolated from
abdominal cramps, low grade fever or an CSF, wounds and blood
absence of fever ● Foodstuffs such as raw milk and beef
● Stool contains no leukocytes E.vulneris
- Distinguishing characteristic from ● Yellow pigmented colonies (more than half
Shigella spp. and EIEC infections of strains); isolated from humans with
● Processed meats (undercooked infected wounds
hamburgers), unpasteurized dairy E.albertii
products, apple cider, bean sprouts and ● associated with diarrheal disease in children
spinach
● Produces two cytotoxins (Shiga like toxins) Klebsiella
verotoxin I (Stx1) and verotoxin II (Stx2) ● Found in the intestinal tract of humans
● Primary screening test Stool culture using and animals or free living in soil, water,
MAC containing sorbitol (SMAC) and on plants
- Non sorbitol fermenter E. coli ● Associated with various opportunistic and
O157:H7 (colorless) hospital acquired infections, particularly
- Sorbitol fermenter other E. coli pneumonia, wound infections, and UTIs
strains ( pink colonies) Common Characteristics
● Additional test 4 methylumbelliferyl β D ● LF on MAC
glucuronide (MUG) assay ● Most grow on SCA and in Potassium
● E. coli O157:H7 rarely produces the Cyanide Broth (KCN)
enzyme β glucuronidase ● H2S negative
● Other strains producer ● A few hydrolyze urea slowly
● (+) Fluorescence ● All are MR and VP+
Enteroadherent E. coli ● With a few , no indole is produced from
● Associated with two kinds of human tryptophan
disease: diarrheal syndromes and UTIs ● Variable motility
● Two types: DAEC and EADC Species:
● DAEC UTIs and diarrheal disease 1. K. pneumoniae subsp. p neumoniae
- Cystitis in children and acute 2. K. oxytoca ,
pyelonephritis in pregnant women 3. K. pneumoniae subsp. o zaenae
- Chronic or recurring UTIs 4. K. pneumoniae subsp. rhinoscleromatis ,
● EAEC causes diarrhea by adhering to the 5. K. ornitholytica
surface of the intestinal mucosa 6. K. planticola ,
7. K. terrigena
K. pneumoniae (Friedlander’s bacillus) Enterobacter, Cronobacter and Pantoea
● Most commonly isolated specie Clinically significant species:
● Posses a large polysaccharide capsule 1. Enterobacter cloacae
● Responsible for the moist, mucoid colonies 2. Enterobacter aerogenes
characteristic of K. pneumoniae on MAC 3. Enterobacter gergoviae
● Frequent cause of lower respiratory tract 4. Enterobacter hormaechei
infection among hospitalized patients and ● Motile
immunocompromised hosts such as ● MAC colony morphology resembles
newborns, elderly patients and seriously ill Klebsiella
patients of respirators
● Causes community acquired pneumonia Enterobacter, Cronobacter , Pantoae
cough up “currant jelly like” sputum ● E. cloacae and E. aerogenes two most
● Also causes wound infections, UTIs, liver common isolates
abscesses and bacteremia ● Isolated from wounds, urine, blood and CSF
● Plasmid transfer of antimicrobial resistance ● Pantoea (Enterobacter) agglomerans
Carbapenemase nationwide outbreak of septicemia resulting
● Blood tinged or rust colored from contaminated IV fluids
● E. agglomerans complex lysine --,
K. pneumoniae ornithine --, and arginine or “triple
● Virulence factor - polyssacharride capsule decarboxylases negative”
● Differential test - string test ● E. gergoviae --> found in respiratory
● Neufeld Quelleng test: Positive samples and is rarely isolated from blood
● Growth in potassium cyanide (KCN): cultures
positive ● Cronobacter (Enterobacter) sakazakii
● IMViC : ----+ produces a yellow pigment and has been
● TSI: A/A, gas+, H2S documented as a pathogen in neonates
causing meningitis and bacteremia, often
K. oxytoca coming from powdered infant formula
● Produces infections similar to those caused ● E. hormaechei isolated from human
by K. pneumoniae sources such as blood, wounds, and
● Linked to antimicrobial associated sputum.
hemorrhagic colitis ● E. asburiae is similar biochemically to E.
● Identical to K. pneumoniae except for its cloacae and has been isolated from blood,
production of indole, and there are reports urine, feces, sputum, and wounds.
of ornithine positive isolates as well ● E. cancerogenus (formerly E. taylorae )
associated with osteomyelitis after
K. pneumoniae subsp. ozaenae traumatic wounds.
● Isolated from nasal secretions and
cerebral abscesses Enterobacter
● Causes atrophic rhinitis a tissue ● Ornithine decarboxylase (ODC) positive
destructive disease restricted to the nose ● Lysine decarboxylase (LDC) is positive by
(“foul smelling” atrophic rhinitis) most species ( except E. gergoviae and E.
cloacae
K. pneumoniae subsp. rhicoscleromatis ● Growth on KCN agar
● Causes rhinoscleroma - an infection of the ● Sorbitol fermentation +
nasal cavity that manifests as an intense ● IMVic - - ++
swelling and malformation of the entire ● TSI: A/A (except E. agglomerans K/A), gas+
face and neck (except E. agglomerans neg ), H2S
Serratia Proteus Both species are:
Species: ● Urease positive
1. S. marcescens (most clinically significant) ● H2S producer
2. S. liquefaciens P. mirabilis
3. S. Rubidaea - Indole neg; Ornithine pos
4. S. Odorifera P. vulgaris
5. S. Plymuthica - Indole pos; Ornithine neg
6. S. ficaria ● Sucrose fermenter; A/A in TSI
7. S. Entomophila
8. S. fonticola Morganella
● Late LF on MAC except for S. fonticola Only one specie M. morganii
Serratia ● Two subspecies: M. morganii subsp.
● S. marcescens , S. rubidaea , and S. morganii and M. morganii subsp. sibonii
plymuthica produces “ prodigiosin”, pink ● Causes UTI and neonatal sepsis
red pigment ● Motile but does not swarm
S. odorifera Morganella
- musty and pungent odor of “rotten potato ● Same biochemical ID as P. vulgaris (except
like” odor that P. vugaris is citrate +)
S. liquifaciens ● NLF on MAC
- ferments arabinose growth in KCN media ● PAD test +
S. marcescens most significant ● IMViC + + - -
● Causes outbreak in nurseries, burn units ● LIA rxn: R/A
and cardiac surgery units ● TSI rxn : K/A, gas +, H2S
● Urease, gelatinase positive, arabinose ● Urease, KCN, ornithine decarboxylase
fermentation negative positive
General characteristics: ● Motile but does not swarm
● IMViC ----+ Providencia
● TSI rxn : K/A, gas +, H2S Five species:
Hafnia 1. P. alcalifaciens is most commonly found in
● Composed of only one specie: H. alvei the feces of children with diarrhea
● Gastroenteritis 2. P. stuartii outbreaks in burn units; isolated
● Major characteristic: delayed positive from urine culture
citrate reaction 3. P. rettgeri documented pathogen of urinary
Proteus tract; also diarrheal disease among
● P. vulgaris and P. mirabilis widely travelers
recognized human pathogens 4. P. rustigianii formerly identified as a strain
- P. mirabilis most common clinical of P. alcalifaciens
isolate 5. P. heimbachae
● Disseminated in the environment, are
normal intestinal microbiota, and are Edwardsiella
recognized as opportunistic pathogens Three species:
● Distinguished from the other members of E. tarda only recognized human pathogen
the Enterobacteriaceae by virtue of the - Causes bacteremia and wound infections
ability to deaminate the amino acid E. hoshinae
phenylalanine E. ictaluri
● NLF ● NLF in MAC, urease test
● Urease activity of P. mirabilis can lead to ● Positive for lysine decarboxylase (LDC)
struvite kidney stones (calculi) ● IMViC + + - -
● Can produce “swarming” colonies on ● TSI rxn: K/A, gas +, H2S +
nonselective media, such as SBA
● “Burnt chocolate” odor
Proximal kidney tubules --Acute glomerulonephritis
Citrobacter ● Most common mode of transmission
● Consist of 11 species that all have been ingestion of contaminated water or food
isolated from clinical specimens (uncooked or undercooked seafood such as
- Most often isolated are C. freundii , oysters, clams, or shrimp)
C. koseri , and C. braakii
● Produces colonies on MAC that resemble Plesiomonas
E. coli Three major clinical types of gastroenteritis
- Citrobacter Late LF 1. The more common watery or secretory
● Considered as inhabitants of the GI tract diarrhea
and are associated with hospital acquired 2. A subacute or chronic disease that lasts
infections most frequently UTIs from 14 days to 2 to 3 months
● C. freundii has been associated with 3. A more invasive, dysenteric form that
infectious diseases acquired in hospital resembles colitis
settings: UTIs, pneumonias and intra ● 25% to 40% of all patients present with
abdominal abscesses fever, vomiting, or both, and the single most
- Also endocarditis in intravenous common clinical symptom for all such
drug abusers patients is abdominal pain
- Because it is H2S positive and 50% ● Self limiting, but antimicrobial therapy is
of the strains fail to ferment lactose indicated in severe and prolonged cases
the colony morphology on selective ● Shown a general resistance to the
media can be mistaken for penicillin class of antibiotics
Salmonella when isolated from stool
cultures ● Vibriostatic test 0/129: Sensitive
1. Differentiated by urea hydrolysis and ● NLF on MAC (but some strain will not grow)
lysine decarboxylase ● Non hemolytic, smooth and opaque in
2. 70% C. freundii urease pos; BAP
Decarboxylate lysine neg ● Inositol brilliant green bile salt agar :
3. Salmonella urease neg, Decarboxylate white or green to pink color enhances the
lysine pos recovery from specimens
Citrobacter ● Growth in HEA and CIN, no growth in
● Urease + TCBS and media with NaCl
● IMViC: TCBS- THIOSULFATE CITRATE BILE SALT
- C. freundii - + - + SUCROSE AGAR
- C. koseri + + - + CIN - CEFSULODIN IRGASAN NOVOBIOCIN
● TSI rxn: AGAR
- C. freundii A/A or K/A, gas +, H2S + Yersinia spp.
- C. koseri K/A, gas +, H2S -
Plesiomonas (Vibrionaceae) Plesiomonas
● Only one specie P. shigelloides ● Oxidase positive
- It is found in soil and aquatic ● Decarboxylase test: positive to trio
environments (fresh and estuarine decarboxylase test
waters of tropical and subtropical ● Inositol fermentation: positive
climates) ● IMViC + +
- Does not tolerate increased NaCl ● TSI rxn : K/A, gas --, H2S
- Minimum growth temperature of 8 C ● Antigenic structure: O and H antigens
● Widely distributed among warm and cold
blooded animals
- Dogs, cats, vultures, snakes, lizards,
fish, newts, and shellfish
● Similar colonies with E. coli on ordinary
enteric media
● Also a potential cause of enteric disease
Primary Intestinal Pathogens of the Family ● S. bongori is a rarely isolated species
Enterobacteriaceae named after the town of Bongor in Chad,
Salmonella , Shigella and Yersinia Africa
● Salmonella and Shigella organisms produce ● S. enteritidis, S. cholerasuis and S. typhi
GI illnesses in humans not considered ● S. enterica subsp. Enterica serotype Typhi
normal biota of the human intestinal tract or Salmonella Typhi
inhabit the GI tracts of animals Salmonella
● Humans acquire the infection by ingesting Antigenic Structures
the organisms in contaminated animal ● Somatic O antigen (heat stable) and
food products or insufficiently cooked flagellar H (heat labile) antigen--primary;
poultry, milk, eggs, and dairy products used for serologic grouping
● Some Salmonella infections are ● Capsular K antigen (Vi antigen)-- few
transmitted by human carriers strains; prevents phagocytosis
● Shigella dysentery usually indicates Virulence Factors
improper sanitary conditions and poor ● Fimbriae role in adherence in initiating
personal hygiene intestinal infection
● Yersinia infections include GI disease, ● Enterotoxin causes gastroenteritis
mediastinal lymphadenitis, fulminant Clinical Infections (Salmonellosis)
septicemia, and pneumonia 1. Acute gastroenteritis or food poisoning
vomiting and diarrhea
Serotype or serovar 2. Typhoid fever most severe form of enteric
● Are groups within a single species of fever
microorganisms which share distinctive ● Caused by Salmonella serotype Typhi
surface structures 3. Enteric fevers
● Type and number of cell surface Antigens ● Caused by other Salmonella serotypes
● Share similar antigen (antigenically similar) (e.g., Salmonella Paratyphi and
Salmonella Choleraesuis
● Found in cold blooded animals as well as 4. Nontyphoidal bacteremia
in rodents and birds (natural host) 5. Carrier state following Salmonella infection
● Most pathogenic Enterobacteriaceae ● Humans acquire the infection by ingesting
● Causes enteric fever (typhoid fever) and the organisms in food, water, and milk
acute gastroenteritis (food poisoning) contaminated with human or animal
● MAC produces clear, colorless, non lactose excreta
fermenting colonies ● Salmonella serotypes Typhi and
● HEA and XLD colonies with black centers Paratyphi no known animal reservoirs;
are seen infections only occur in humans
Salmonella - Biochemical Features Salmonella Gastroenteritis
● NLF ● most common forms of “food poisoning”
● IMVIC - + - - ● GI infection caused by salmonellae results
● Phenyalanine and Urease Neg from the ingestion of the organisms through
● H2S Producer (except S. paratyphi A) contaminated food
● Do not grow in KCN medium ● Source of infection poultry, milk, eggs and
Salmonella egg products as well as to handling pets
S. enterica - type specie of the genus - Common insufficiently cooked aggs and
Subspecies: domestic fowl, such as chicken, turkey and
● S. enterica subsp. enterica (subspecies I) duck
● S. enterica subsp. salamae (subspecies II) - Cooking utensils, such as knives, pans, and
● S. enterica subsp. arizonae (subspecies IIIa cutting boards used in preparing the
● S. enterica subsp. diarizonae (subspecies contaminated meat, can spread the bacteria
(IIIb) to other food
● S. enterica subsp. houtenae (subspecies IV) ● Direct transmission from person to person
● S. enterica subsp. Indica (subspecies VI) ● Infective dose 10^6 bacteria
 ● Symptoms may appear 8 to 36 hours after Salmonella Bacteremia
ingestion ● The serotypes most commonly associated
● Nausea, vomiting, fever, and chills, with bacteremia are Typhimurium, Paratyphi
accompanied by watery diarrhea and , and Choleraesuis (nontyphoidal
abdominal pain Salmonella)
● Most cases of Salmonella gastroenteritis are ● Observed in two different groups:
self limiting 1. Young children, who experience fever and
● Susceptible to infection patients with gastroenteritis with brief episodes of
sickle cell disease and other hemolytic bacteremia
disorders, ulcerative colitis and malignancy 2. Adults, who experience transient
● Antimicrobials of choice include bacteremia during episodes of
chloramphenicol, ampicillin, and gastroenteritis or develop symptoms of
trimethoprim sulfamethoxazole septicemia without gastroenteritis
Salmonella Typhoid Fever Carrier State
● Enteric fever caused by Salmonella Typhi is ● Who recover from infection may harbor the
known as typhoid fever organisms in the gallbladder, excreted in
● A febrile disease that results from the feces
ingestion of food contaminated with the ● Antimicrobial therapy if gallbladder
organisms originating from infected infection is not evident
individuals or carriers ● Cholecystectomy has been the only
Clinical features: solution to the chronic state of enteric
● Prolonged fever carriers
● Bacteremia Shigella
● Involvement of the reticuloendothelial ● Named after the Japanese microbiologist
system, particularly the liver, spleen, Kiyoshi Shiga who first isolated the
intestines, and mesentery organism in 1896
● Dissemination to multiple organs ● Closely related with Escherichia
● No known animal reservoir ● Not a member of the normal GI microbiota
● Paratyphoid fevers caused by Salmonella all spp. can cause bacillary dysentery
serotypes Paratyphi A, B, and C and ● Species: S. dysenteriae , S. flexneri , S.
Salmonella serotype Choleraesuis --- less boydi , S. sonnei
severe ● S. dysenteriae most virulent
● Typhoid fever develops approximately 9 to ● S. flexneri one of the causes of gay bowel
14 days after ingestion of the organisms syndrome
● The onset of symptoms depends on the Characteristics:
number of organisms ingested the larger the 1. NLF (except S. sonnei )
inoculum, the shorter the incubation period 2. Nonmotile
● First week of disease fever accompanied 3. Produce gas from glucose (except S.
by malaise, anorexia, lethargy, myalgia, and flexneri
a continuous dull frontal headache 4. Urease neg
● Second and third week of disease 5. Does not produce H2S
sustained fever with prolonged bacteremia 6. They do not decarboxylate lysine
- 2nd week ––“Rose spots” ● Serogroups: A ( S. dysenteriae ), B S.
(blanching, rose colored papules flexneri ), C S. boydi ), D S. sonnei
around the umbilical region) appear ● Antigenic structure: Somatic O
● Specimen: stool or rectal swab
● Sensitive to pH change, must be process
immediately in lab
Shigella Clinical Infections and Identification organisms proliferate in the bloodstream
S. dysenteriae - causes the enteric disease and respiratory tract
bacillary dysentery ● Can be a primary infection when the
● Characterized by the presence of blood, bacteria is inhaled
mucus, and pus in the stool Yersinia pestis
● IMViC v + - - ● Gram negative, short, plump bacillus
● TSI rxn : K/A, gas --, H2S ● Class A bioterrorism agent
S. sonnei - unique in its ability to decarboxylate ● Methylene blue or wayson stain shows
ornithine intense staining at each end of the bacillus
● Produces a “delayed” positive fermentation referred to as the bipolar staining “safety
of lactose with the formation of pink colonies pin” appearance
on MAC agar only after 48 hours of ● Preferred growth temperature 25 C to 30 C
incubation ● NLF in MAC, pinpoint at 24hrs in BAP
● ONPG positive ● “Stalactite shaped” pattern in broth
● Self limiting infection characterized by fever, ● IMViC - + - -
watery diarrhea without blood ● TSI rxn K/A. gas --, H2S
● Humans are the only known reservoir Yersinia enterocolitica
● Shigellosis is highly communicable ● Most commonly isolated specie
- Low infective dose is required to ● Can be acquired from contact with
produce the disease (<100 bacilli are household pets
needed to initiate the disease) ● Causes enterocolitis or waterborne
Mode of transmission: gastroenteritis
1. Direct person to person contact - Stools may contain blood
● Via fecal oral route - Mild and self limiting
● Anal oral sexual activity ● Associated with the transfusion of
● Anal intercourse (men to men) S. flexneri contaminated packed red blood cells
2. Transmitted by flies, fingers and food or (sepsis) survives in cold temperature
water contaminated by infected persons ● Related infections appendicitis like
Yersinia syndrome, arthritis, and erythema nodosum
Three species (human pathogen): ● Appendicitis like syndrome severe
● Y. pestis causative agent of plague, a abdominal pain that is concentrated in the
disease primarily of rodents transmitted to right lower quadrant and fever
humans by fleas ● Arthritis common extraintestinal form of Y.
● Y. pseudotuberculosis and Y. enterocolitica infection
enterocolitica caused sporadic cases of ● Erythema nodosum inflammatory reaction
mesenteric lymphadenitis in humans, characterized by tender, red nodules that
especially in children, and generalized may be accompanied by itching and burning
septicemic infections in ● Acquired thru ingestion of undercooked food
immunocompromised hosts (pork, pork intestines and vacuum packed
● Y. enterocolitica produces an infection that meat), dairy products (chocolate milk) and
mimics appendicitis handling of pets
Three forms of plague in humans: ● Gram negative coccobacilli with bipolar
1. Bubonic or glandular form most common; staining
results from the bite of an infected flea ● Optimal growth temperature 25 C to 30 C
- High fever with painful regional lymph ● Motile @ 25 C but not at 35 C
nodes known as buboes (swollen ● CIN ( Cefsulodin irgasan novobiocin) agar
lymph nodes) selective medium with bull’s eye
2. Septicemic form bacteria spread to the appearance or dark red or burgundy
bloodstream center transparent borders” @ 48 hrs
3. Pneumonic form occurs secondary to ● Inhibitory agents - cefsulodin , irgasan ,
bubonic or septicemic plague when novobiocin, bile salts, and crystal violet
● Inhibits normal colon microbiota
 ● MAC is NLF
● IMViC v + - -
● TSI rxn : K/A, gas --, H2S
Yersinia pseudotuberculosis
● Pathogen of rodents, particularly guinea
pigs
● Reservoir are farm and domestic animals ,
usually birds
● Causes a disease characterized by caseous
swellings called pseudo tubercles fatal in
animals
● Human infections are rare
● To differentiate from Y. pestis motile @ 18
C to 22 C, urease positive and rhamnose
fermentation
● MAC is NLF
● TSI rxn : K/A, gas --, H2S