BACT211: CLINICAL BACTERIOLOGY (Lecture)

WEEK 13: “Enteric Gram-Negative Rods (Enterobacteriaceae)”

2nd Semester | S.Y. 2022-2023
Professor | Ms. Rochelle D. Darlucio

GENERAL CHARACTERISTICS VI. Proteeae Proteus mirabilis

• Often referred to as enterics or coliforms vulgaris
penneri
• Gram-negative bacilli and coccobacilli hauseri
• Facultative anaerobes or aerobes myxofaciens
• Do not produce cytochrome oxidase EXCEPT for Morganella morganii
Plesiomonas Providencia alcalifaciens
• Ferment a wide range of carbohydrates (all ferment rettgeri
glucose) stuartii
VII. Yersinieae Yersinia pseudotuberculosis
• Reduce nitrate to nitrite EXCEPT for Photorhabdus and pestis
Xenorhabdus enterocolitica
• Motile (peritrichous flagella) at body temperatures frederiksenii
EXCEPT for Klebsiella, Shigella, and Yersinia kristensenii
• Catalase positive EXCEPT Shigella dysenteriae type 1 intermedia
ruckeri
• Possess a complex antigenic structure
• Produce a variety of toxins and other virulence factors • H2S – blackening of the colony or medium
o Observed on Triple Sugar Iron agar or TSI
• Large heterogenous group of gram-negative rods in which ▪ Acronym – SPACE
their natural habitat is in the intestinal tract of humans and • Salmonella
animals • Proteus
• Surface of BAP/CAP – appear as large, moist, gray color • Arizona
• Tribes – classify the members of the family • Citrobacter
o Proposed by Ewing in 1963
• Edwardsiella
o Grouped bacterial species with similar biochemical
o Observed on Lysine Iron Agar or LIA
characteristics
▪ Acronym – SACE
o Within tribes, classified into genus and species
• Salmonella
TABLE 19.1 Classification of Selected Species Within the Family • Arizona
Enterobacteriaceae • Citrobacter
TRIBE GENUS SPECIES • Edwardsiella
I. Escherichieae Escherichia coli • Urease
albertii o Rapid Urease Producers
blattae ▪ PPM
vulneris
fergusonii • Proteus
hermanii • Providencia
Shigella dysenteriae • Morganella
flexneri o Slow Urease Producers
boydii ▪ CKEYS
sonnei
• Citrobacter
II. Edwardsielleae Edwardsiella tarda
liquefaciens • Klebsiella
hoshinae • Enterobacter except. E. gergoviae
ictaluri • Yersinia
III. Salmonelleae Salmonella enterica • Serratia
bongori • Deaminase Producing
IV. Citrobacteriaceae Citrobacter freundii
o PPM
koseri (C. diversus)
amalonaticus ▪ Proteus
youngae ▪ Providencia
braakii ▪ Morganella
farmeri • Lysine Decarboxylation (LDC) Positive
V. Klebsielleae Klebsiella pneumoniae subsp. o KEESSH
pneumoniae ▪ Klebsiella
pneumoniae subsp.
▪ Escherichia
ozaenae
Enterobacter aerogenes
▪ Edwardsiella
cloacae ▪ Serratia
gergoviae ▪ Salmonella except S. paratyphi A
cancerogenus ▪ Hafnia
(taylorae) • Lysine Decarboxylation (LDC) Negative
hormaechei o PPMCYES
Pantoea agglomerans ▪ Proteus
Cronobacter sakazakii
▪ Providencia
Hafnia alvei
▪ Morganella
Serratia marcescens
liquefaciens ▪ Citrobacter
rubidaea ▪ Yersinia
fonticola ▪ Enterobacter except E. aerogenes and E.
odorifera gergoviae
plymuthica ▪ Shigella

Page 1 | W e e k 1 3
 WEEK 13: “Enterobacteriaceae”

TABLE 19.2 Biochemical Characteristics of Tribes of Enterobacteriaceae

TESTS or Escherichieae Edwardsielleae Citrobacteriaceae Salmonelleae Klebsiellae Proteeae Yersiniae
SUBSTRATE
H2S (TSI agar) - + + or - + - + or - -
Urease - - (+w) or - - - or (+) + or - +
Indole + or - + - or + - - + or - + or -
Methyl red + + + + - + +
Voges-Proskauer - - - - + - -
Citrate (Simmons) - - + + + d -
KCN - - + or - - + + -
Phenylalanine - - - - - + -
deaminase
Mucate d - d + or - -
Mannitol + or - - - + + - or + +

PATHOGENESIS AND SPECTRUM OF DISEASES • Facultative anaerobic, glucose fermenting, gram-negative,

1. Opportunistic Pathogens oxidase-negative rods capable of growth on MacConkey
o Part of the intestinal microbiota of animals and agar
humans • Used as a primary marker of fecal contamination in
o Outside normal body sites, able to produce serious water quality testing
extraintestinal opportunistic infections • Motile and generally possess adhesive fimbriae and sex
o Ex. E. coli pili and O, H, and K antigens
▪ Member of the normal bowel biota • Lactose-positive (pink) colony with a surrounding area of
▪ Normally present in feces precipitated bile salts on MAC agar – lactose fermenter
▪ On other body parts, can cause: • EMB Agar – green metallic sheen
• UTI,
• septicemia, • Urinary tract infections, bacteremia, neonatal meningitis,
• meningitis on neonates and nosocomial infections of other various body sites
o Other examples • Most common cause of gram-negative nosocomial
▪ Citrobacter infections
▪ Enterobacter
▪ Klebsiella PROPERTIES:
▪ Proteus • Fermentation of glucose, lactose, trehalose, and xylose
▪ Serratia spp. • Production of indole from tryptophan
2. Intestinal Pathogens
• Glucose fermentation by the mixed acid pathway: methyl
o Referred to as primary pathogens red positive and Voges-Proskauer negative
o Examples • Does not produce H2S (no blackening),
▪ Salmonella typhi deoxyribonuclease (DNase), urease, or phenylalanine
• Cause typhoid fever or enteric fevers deaminase
▪ Shigella • Cannot use citrate as a sole carbon source
• Cause dysentery
Uropathogenic Escherichia Coli
VIRULENCE AND ANTIGENIC FACTORS
• O antigen, or somatic antigen • Most common cause of UTIs in humans
o Heat-stable antigen • Strains allow them to attach to epithelial urinary mucosa
o Located on the cell wall
• H antigen, or flagellar antigen Gastrointestinal Pathogens
o Heat-labile antigen • Cause diarrhea
o Found on the surface of the flagella (responsible for • Enterovirulent E. coli or diarrheogenic E. coli:
motility o Enterotoxigenic Escherichia coli (ETEC)
• K antigen, or capsular antigen o Enteroinvasive Escherichia coli (EIEC)
o Heat labile polysaccharide o Enteropathogenic Escherichia coli (EPEC)
o Often found on encapsulated species o Enterohemorrhagic Escherichia coli (EHEC)
o Example o Enteroadherent
▪ E. coli – K1 antigen ▪ Diffusely Adherent Escherichia coli (DAEC)
▪ Salmonella enterica – Vi antigen (Vi = virulence) ▪ Enteroaggregative Escherichia coli (EAEC)

OPPORTUNISTIC MEMBERS OF THE FAMILY Enterotoxigenic Escherichia coli (ETEC)

ENTEROBACTERIACEAE AND ASSOCIATED • diarrhea of adults and especially children
INFECTIONS • one of the major causes of infant bacterial diarrhea
A. Escherichia coli (IMVC ++--) • most common cause of a diarrheal disease sometimes
• IMVC – Indole (+), Methyl Red (+), Voges-Proskauer (-), referred to as traveler’s diarrhea
Citrate (-) • MOT: consumption of contaminated food or water
• 1885 – Escherich • Achlorhydria: high risk factor
• Colon bacillus or Golden Bacillus o Deficiency of hydrochloric acid in the stomach
• produces heat-labile enterotoxin
• watery diarrhea, abdominal cramps, and sometimes
nausea, usually with no vomiting or fever
Page 2 | W e e k 1 3
 WEEK 13: “Enterobacteriaceae”

Enteropathogenic Escherichia coli (EPEC) B. Klebsiella and Raoultella (IMVC --++)

• known to cause infantile diarrhea • Indole (-), Methyl red (-), Voges-Proskauer (+), Citrate (+)
• characterized by low-grade fever, malaise, vomiting, • Most grow on Simmons Citrate and in Potassium Cyanide
and diarrhea Broth
• O serogroups – cause of diarrhea • None produce H2S
• H antigenic – intestinal infections • A few hydrolyze urea slowly
• STOOL: watery with mucus but without blood • All are methyl red test negative and Voges-Proskauer
Enteroinvasive Escherichia coli (EIEC) positive
• produce dysentery with direct penetration, invasion, • With a few exceptions, no indole is produced from
and destruction of the intestinal mucosa tryptophan
• similar to shigellosis • Motility is variable
• MOT: person to person via the fecaloral route • Members can be found on the intestinal tract of humans
and animals
• nonmotile and generally do not ferment lactose -NLF • Associated with opportunistic and hospital-acquired
• do not decarboxylate lysine infections
Enterohemorrhagic Escherichia coli (EHEC) o Pneumonia
• O157:H7 - hemorrhagic diarrhea, colitis, and hemolytic o Wound infections
uremic syndrome (HUS) o UTI
o First recognized during the outbreak of • K. pneumoniae - most commonly isolated species; large
hemorrhagic diarrhea and colitis polysaccharide capsule
o HUS – characterized by low platelet count and o Capsule – provide protection phagocytosis;
presence of hemolytic anemia and kidney failure responsible for moist, mucoid colonies
o does not ferment sorbitol in 24 hours o Capsulated: mucoid colonies or tend to “string”
o Biochemical test: 4-methylumbelliferyl-β-D- o Friedlander’s Bacillus
glucuronide (MUG) assay o Cause pneumonia – very necrotic and hemorrhagic –
▪ O157:H7 Rarely produce β-D-glucuronidase “currant jelly like sputum”
but 92% of strains able to produce
▪ If with enzyme – MUG will be cleaved and a Klebsiella oxytoca
fluorescent product is formed (fluorescence) • the only indole positive Klebsiella species
o 2 toxins: • isolated in stool and blood cultures
▪ Verotoxin I – identical to Shiga toxin • ornithine-positive
(produced by Shigella dysenteriae); produce • linked to antibiotic-associated hemorrhagic colitis
damage to Vero cells (African green monkey K. pneumoniae subsp. ozaenae
kidney cells) • highly associated with the presence of plasmid-
▪ Verotoxin II – Shiga-like toxin; most likely to mediated ESBLs
be found on Shiga toxin 1 and Shiga toxin 2
• isolated from nasal secretions and cerebral abscesses
(Shiga-toxin producing E. coli [STEC])
K. pneumoniae subsp. rhinoscleromatis
• watery diarrhea
• has been isolated from patients with rhinoscleroma
• bloody diarrhea with abdominal cramps and low-grade
(infection of the nasal cavity – intense swelling and
feer or an absence of fever
malformation of the entire face and neck)
Enteroadherent Escherichia coli
Raoutella (Klebsiella) ornithinolytica
• diarrheal syndromes and UTIs – DAEC (Diffusely
• indole and ornithine decarboxylase-positive
Adherent E. coli)
Raoutella (Klebsiella) planticola
• EAEC (Enteroaggregative E. coli) – diarrhea by
adhering to the surface of the intestinal mucosa • have been isolated from the urine, respiratory tracts,
and blood of humans
Extraintestinal Infections
K. variicola
• most common causes of septicemia and meningitis
among neonates • has been isolated from primarily sterile sites
OTHER ESCHERICHIA SPECIES Klebsiella granulomatis (formerly Calymmatobacterium
granulomatis)
Escherichia hermannii
• yellow-pigmented • granuloma inguinale
• a sexually transmitted disease
• isolated from cerebrospinal fluid (CSF), wounds, and
blood
Escherichia vulneris
• isolated from humans with infected wounds
• yellow pigmented
Escherichia albertii
• associated with diarrheal disease in children

Page 3 | W e e k 1 3
 WEEK 13: “Enterobacteriaceae”

Table 19.5 Differentiation of Common Species Within the Genus Klebsiella

K. pneumoniae subsp. pneumoniae K. oxytoca K. pneumoniae subsp. ozaenae
Test or Substrate Sign %+ (% +) Sign %+ (% +) Sign %+ (% +)
Urease + 95.4 (0.1) + 90 d 0 (14.8)
Indole - 0 + 99 - 0
Methyl red - or + 10 - 20 + 97.7
Voges-Proskauer + 98 + 96 - 0
Citrate (Simmons) + 98 (0.6) + 95 d 30 (32.4)
Gelatin (22’C) - 0 (0.2) - 0 - 0
Lysine decarboxylase + 98 (0.1) + 99 - or + 40 (6.3)
Malonate + 92.5 + 98 - 6
Mucate + 90 + 93 - or + 25
Sodium alginate + or 88.5 (9.2) nd - or (+) 0 (11)
(utilization) (+)
Gas from glucose + 96 + 97 d 50 (9.4)
Lactose + 98.7 (1) + 100 d 30 (61.3)
Dulcitol - or + 30 + or - 55 - 0
Organic acid media
Citrate + or - 64.4 nd - or + 18
D-Tartrate + or - 67.1 nd - or + 39

Enterobacter, Cronobacter, and Pantoea (IMVC --++) S. odorifera

• Indole (-), Methyl red (-), Voges-Proskauer (+), Citrate (+) • Dirty, musty odor resembling that of rotten potatoes
• Motile
• Resembles Klebsiella when growing on MAC agar • Two biogroups:
• Grow on Simmons Citrate medium and in Potassium o S. odorifera biogroup 1
Cyanide broth ▪ isolated predominantly from respiratory tract
• The methyl red test is negative, and the Voges-Proskauer ▪ positive for sucrose, raffinose, and ornithine
test is positive ▪ indole-positive (60%)
• Usually produce ornithine decarboxylase
• Lysine decarboxylase is produced by most species but not o S. odorifera biogroup 2
by E. gergoviae or E. cloacae ▪ negative for sucrose, raffinose, and ornithine
• Enterobacter – slow urease producer EXCEPT for E. ▪ isolated from blood and CSF
gergoviae ▪ may also be indole-positive (50%)
Serratia liquefaciens
Enterobacter taylorae • very similar to S. marcescens but differentiated to S.
• lactose negative but ONPG positive marcescens by its ability to ferment arabinose
Enterobacter gergoviae Serratia rubidaea
• resembles Enterobacter aerogenes • also produces red pigment - prodigiosin
• Rapid Urease Producer (RUP)
• causes infection of the urinary tract, and blood Hafnia (IMVC -V+--)
Cronobacter (Enterobacter) sakazakii • Indole (-), Methyl red (variable), Voges-Proskauer (+),
• biochemically similar to Enterobacter cloacae Citrate (-)
• typically produces a yellow pigment • Has been linked to gastroenteritis
• pathogen in neonates – meningitis and bacteremia, • Occasionally isolated from stool cultures
often coming from powdered infant formula • Delayed positive citrate reaction
Pantoea (Enterobacter) agglomerans • Motile and Late Lactose Fermenter (LLF)
• lysine-, ornithine-, and arginine-negative or triple • ONPG positive
decarboxylases negative • Lysine decarboxylase (LDC) positive
• indole positive and mannitol, raffinose, salicin, sucrose, • Ornithine decarboxylase (ODC) positive
maltose, and xylose negative
• Hafnia alvei
Serratia (IMVC -V++) o Formerly Enterobacter alvei
o Resembles Enterobacter. To differentiate:
• Indole (-), Methyl red (variable), Voges-Proskauer (+),
▪ Late lactose fermenter
Citrate (+)
▪ Citrate negative (-)
• Opportunistic pathogens
o To differentiate from Serratia:
• Associated with outbreaks in health care settings – urinary ▪ DNAse negative
or respiratory tract infections ▪ Lipase negative
• Ferment lactose slowly (LLF) and are positive for the o- ▪ Gelatinase negative
nitrophenyl-β-D-galactopyranoside (ONPG) test EXCEPT
S. fonticola
Proteus
• Ability to produce extracellular DNase
• Most of them are classified as non-pigmented • Tribe Proteeae – ability to deaminate the amino acid
phenylalanine; non lactose fermenters
S. marcescens produce a characteristic pink-to-red • Normal intestinal microbiota; opportunistic pathogens
S. rubidaea pigment, prodigiosin, especially when the • “chocolate cake” or “burnt chocolate” smell
S. plymuthica cultures are incubated at room temperature

Page 4 | W e e k 1 3
 WEEK 13: “Enterobacteriaceae”

• Source of antigen for Weil-felix reaction (serologic test for • E. tarda

diagnosis of Rickettsial diseases) o Opportunist
o Proteus antigens cross-react with Rickettsial antigens o Causing bacteremia and wound infections
o Same common polysaccharide • E. hoshinae
o P. vulgaris – source of OX2 and OX19 antigen o Isolated from snakes, birds, and water
o P. mirabilis – source of OXK (Kingsbury Strain) • E. ictaluri
o Causes enteric septicemia in fish
UNIQUE CHARACTERISTICS:
• Rapid Urease Producers (RUP-RXN within 4 hours) Citrobacter
• Typical swarming motility (growth in waves or swarms) • Hydrolyze urea slowly and ferment lactose (LLF)
o On the surface of BAP • Producing colonies on MAC agar that resemble those of E.
o If inoculated into center, after incubation there is a coli
presence of waves • Grow on Simmons citrate medium
• H2S producing in TSI but not in LIA • Positive methyl red test
• Inhabitants with GI tract and associated with hospital
Proteus mirabilis & Proteus vulgaris acquired infection (UTI)
• Hydrolyze urea and produce H2S – blackening of medium
• P. mirabilis does not produce indole from tryptophan and • C. freundii
is ornithine-positive o Produce H2S and some strains (50%) fail to ferment
• P. vulgaris produces indole and is ornithine-negative; lactose
ferments sucrose; acid/acid reaction in triple sugar iron o Colony morphology on primary selective media can
(TSI) agar be mistaken for Salmonella when isolated from stool
cultures
Morganella o Hydrolyze urea, but all fail to decarboxylate lysine
• C. koseri
• Found in the environment and intestinal tract of humans
o Cause of nursery outbreaks of neonatal meningitis
and animals
and brain abscesses
• Frequently cause nosocomial infections, UTIs, and wound
• C. braakii
infections
o Rare human pathogen
• M. morganii
o Associated with community-acquired infections
o Cause of UTI
including a septicemia in a patient with cervical cancer
o Cause of neonatal sepsis
o Motile but does not swarm
o Indole, methyl red, urease, Ornithine decarboxylase Plesiomonas
and phenylalanine deaminase POSITIVE • Formerly in the family Vibrionaceae
• Oxidase-positive, glucose-fermenting, facultatively
Providencia anaerobic, gram-negative bacilli
• Does not have the ability to produce gas from glcose
POSITIVE NEGATIVE • Susceptible to agent O/129
Deaminase Lactose • Motile by monotrichous or two to five lophotrichous
H2S flagella
• Motile but no swarming
• P. shigelloides
• Ferments mannose and citrate
o Most common watery or secretory diarrhea
• All are normal intestinal microbiota but cause UTI and
o A subacute or chronic disease that lasts from 14 days
resistant to antimicrobial therapy
to 2 to 3 months
o A more invasive, dysenteric form that resembles
P. rettgeri colitis
• pathogen of the urinary tract
• caused occasional outbreaks in healthcare settings PRIMARY INTESTINAL PATHOGENS OF THE
• diarrheal disease among travelers FAMILY ENTEROBACTERIACEAE
P. stuartii
• outbreaks in burn units
Salmonella
• isolated from urine cultures
• Gram-negative, facultatively anaerobic bacilli that
P. alcalifaciens
morphologically resemble other enteric bacteria
most commonly found in the feces of children with diarrhea
• Selective and differential media: clear, colorless, non-
P. rustigianni
lactose-fermenting colonies
• rarely isolated
• Colonies with black centers as seen if the media (e.g. HE
P. heimbachae or XLD) contain indicators for H2S production
• yet to be isolate from any clinical specimens • Do not ferment lactose
• Negative for indole, the Voges-Proskauer test,
Edwardsiella phenylalanine deaminase, and urea
• Negative for urea • Most produce H2S EXCEPT Salmonella paratyphi A
• Positive for lysine decarboxylase, H2S, and indole • Do not grow in medium containing potassium cyanide
• Do not grow on Simmons Citrate
• Associated with water-harboring fish or turtle
• ICP can develop serious wound infections

Page 5 | W e e k 1 3
 WEEK 13: “Enterobacteriaceae”

S. enterica are six subspecies: Table 19.9 Biochemical and Serologic Differentiation of
• S. enterica subsp. enterica (subspecies I) Shigella Species
• S. enterica subsp. salamae (subspecies II) Test S. S. S. S.
• S. enterica subsp. arizonae (subspecies IIIa) dysenteria flexner boydi sonne
• S. enterica subsp. diarizonae (subspecies IIIb) e i i i
• S. enterica subsp. houtenae (subspecies IV) Mannitol - + + +
• S. enterica subsp. indica (subspecies VI) fermentation
ONPG v - v +
VIRULENCE FACTORS: Ornithine - - - +
decarboxylas
• Fimbriae
e
o Adherence to mucosa and initiate intestinal infections
Serogroup A B C D
• Enterotoxin

ANTIGENIC STRUCTURES: Yersinia

• Somatic O antigens • Short, pleomorphic Gram-negative rods that often exhibit
• Flagellar H antigens bipolar staining [use methylene blue or Wayson stain]
• Capsular (K) antigens – Vi antigen [heat-labile] on S. typhi • Catalase positive
• Microaerophilic or facultatively anaerobic
CLINICAL INFECTIONS: • Y. pestis – cause of plague
• Acute gastroenteritis or food poisoning characterized • Y. enterocolitica and Y. pseudotubercolosis – zoonotic
by vomiting and diarrhea food-borne pathogens
o Both typically causing a mild diarrheal disease,
• Typhoid fever – the most severe form of enteric fever,
following ingestion of contaminated food and/or water
caused by Salmonella serotype Typhi
• Enteric fevers – caused by other Salmonella serotypes
(e.g. Salmonella Paratyphi and Choleraesuis) Yersinia pestis
• Nontyphoidal bacteremia • Causative agent of the ancient disease plague
• Carrier state following Salmonella infection – individual o Plague – disease of rodents transmitted to humans
recovered from infection but can harbor the organism • A gram-negative, short, plump bacillus
inside the gallbladder (site of chronic carriage) • Bipolar staining (Methylene blue or Wayson stain) –
“safety-pin appearance”
• Humans are able to acquire infection through ingestion of • May be isolated on routine culture medium
organism found on food, milk, water contaminated with • Grows at 37’C
human or animal excreta • Preferential growth temperature: 25’C to 30’C
3 FORMS
Shigella Bubonic Form
• Kiyoshi Shiga – first isolated Shigella • most common
• Nonmotile • results from the bite of an infected flea
• Do not produce gas from glucose EXCEPT: S. flexneri • symptoms appear 2 to 5 days after infection
• Do not hydrolyze urea • high fever with painful regional lymph
• Do not produce H2S • buboes (swollen lymph nodes) begin to appear
• Do not decarboxylate lysine Septicemia Form
• Do not use acetate or mucate as a source of carbon • occurs when the bacteria spread to the bloodstream
• With O and K antigens Pneumonic Plague
• occurs secondary to bubonic plague or septicemic form
• S. sonnei • can be a primary infection if the bacteria are inhaled
o Ability to decarboxylate ornithine
o Slowly ferments lactose Yersinia enterocolitica
o ONPG-positive
• Gram-negative coccobacilli with bipolar staining
• Shigella dysenteriae – enteric disease (bacillary
dysentery) • Grows on SBA and MAC
• Optimal growth temperature of 25’C to 30’C
CLINICAL INFECTIONS: • Motile at 25’C but not at 35’C
• Found in a wide variety of animals, including domestic
• Shigellosis/Bacillary Dysentery
swine, cats and dogs
o Early Stage
▪ Incubation period for 1-7 days • Infections most likely occur after ingestion of contaminated
▪ Fever, abdominal cramping and pain, diarrhea food (beef, chicken, lamb, milk, water)
o Diarrheic Stage • Several forms:
▪ Watery diarrhea for 3 days o Acute enteritis
o Dysenteric Phase Most common form of infection
▪ Frequent stools with blood, pus, and mucus Fever, headache, abdominal pain, diarrhea (bloody
▪ Bacteria had invaded the lining of the GIT stool)
o Appendicitis-like syndrome
o Arthritis
o Erythema nodosum
Inflammation of fat cells under the skin
Red nodules or lumps
• Produce a heat-stable enterotoxin
• Ability to survive in cold temperatures

Page 6 | W e e k 1 3
 WEEK 13: “Enterobacteriaceae”

Cefsulodin-Irgasan-Novobiocin (CIN) agar Leminorella

• selective medium to detect the presence of Y. • L. grimontii and L. richardii
enterocolitica • produce H2S and have shown weak reactions with
• cefsulodin, irgasan, novobiocin, bile salts, and crystal Salmonella antisera
violet as inhibitory agents Moellerella
Yersinia-Selective Agar (YSA) base • M. wisconsensis
• modification of CIN agar • positive for citrate, methyl red, lactose, and sucrose
• added mannitol • negative for lysine, ornithine, arginine decarboxylase,
• fermentation of mannitol: red at the center of the and indole
colony and the bile to precipitate • resembles E. coli growing on enteric media
• nonfermentation of mannitol produces a colorless, • isolated from feces in two cases of diarrhea, infected
translucent colony gallbladders, and a bronchial aspirate
Obesumbacterium
Yersinia pseudotuberculosis • fastidious, slow-growing organisms at 37’C and have
• Pathogen primarily of rodents, particularly guinea pigs not been found in human specimens
• Characterized by caseous swellings called Photorhabdus
pseudotubercles • three species:
• Appears as a typical-looking plague bacillus o P. luminescens
• Motility at 18’C to 22’C o P. asymbiotica
• Production of urease, and ability to ferment rhamnose o P. temperate
• Disease is often fatal in animals • natural habitat: lumen of entomopathogenic nematodes
• strains have occasionally been isolated from human
specimens
OTHER GENERA OF THE FAMILY • strains produce pink-, red-, orange-, yellow-, or green-
pigmented colonies on nutrient agar and especially on
ENTEROBACTERIACEAE
nutrient-rich media, such as trypticase soy agar and
egg yolk agar
Budivicia • negative for nitrate reduction
• Budivicia aquatica Rahnella
• usually found in water; however, they occasionally • Rahnella aquatilis is the name given to a group of
occur in clinical specimens water bacteria that are psychrotolerant, growing at 4’C
Buttiauxella • resemble E. agglomerans
• B. agrestis and B. noackiae – isolated from human • weak phenylalanine deaminase reaction
specimens • negative for potassium cyanide (KCN), gelatin, lysine,
• similar to both Citrobacter and Kluyvera species, but ornithine, and motility; and their lack of yellow
DNA hybridization distinctly differentiates Buttiauxella pigmentation
from both genera Trabulsiella
Cedecea
• isolated from vacuum-cleaner contents on the island
• Five species: of Guam when environmental indoor dirt samples were
o C. davisae being collected
o C. lapagei Yokenella
o C. neteri
• Yokenella regensburgei
o Cedecea species types 3 and 5
• biochemically similar to Hafnia
• Most have been recovered from sputum, blood, and
• negative Voges-Proskauer test results
wounds
• C. davisae is the most commonly isolated species Tatumella
Ewingella • Tatumella ptyseos
• Ewingella americana is the only species • show more biochemical reactions at 25’C than at 35’C
• motile at 25’C but not at 35’C
• come from human blood cultures or respiratory
specimens and exhibit resistance to multiple • demonstrate large 15- to 36-mm zones of inhibition
antimicrobial agents around penicillin disks
Kluyvera • slow-growing, produce tiny colonies, and are relatively
nonreactive in laboratory media
• three closely related species:
o K. ascorbata (the most common clinical isolate)
o K. cryocrescens
o K. georgiana
• found in respiratory, urine, and blood cultures
• most strains are nonpigmented, but occasional isolates
may produce a reddish-blue or violet pigment
• resemble E. coli colonies growing on MAC agar
• K. ascorbata does not ferment glucose at 5’C
• K. cryocrescens ferments glucose
Leclercia
• L. adecarboxylata – have a yellow pigment but only on
initial isolation
• has similar IMViC reactions to E. coli
• negative for lysine and ornithine decarboxylase and
arginine dihydrolase

Page 7 | W e e k 1 3
 WEEK 13: “Enterobacteriaceae”

KEY REACTIONS
ORGANISM PATHOGENICITY POSITIVE NEGATIVE OTHER
Escherichia UTI, septicemia, Lactose, gas, H2S, VP, citrate, • Predominant aerobe in GI tract
coli neonatal sepsis & indole, MR, PD, urease • Most common cause of UTI
meningitis, diarrhea motility • Green metallic sheen on EMB
(some) • Presumptive ID: lactose pos, dry
colony on Mac-Conkey, oxidase
neg, indole pos
• E. coliO157:H7 doesn’t ferment
sorbitol; colorless colonies on
SMAC
Shigella • Dysentery MR Lactose, gas, • Blood, mucus, polys in stool
(shigellosis) H2S, VP, citrate, • Rarely disseminates
• Most PD, urease, • Fragile organism
communicable of motility • S. dysenteriae most severe
bacterial diarrheas • Shigella sonnei most common in
• Found primarily in U.S. Serogrouped by 0 antigens
crowded or • Serogroups A,B,C,D
substandard • Closely related to Escherichia on
conditions, e.g., molecular basis
day-care centers,
jails, prisons
Edwardsiella • Opportunistic Gas, H2S, indole, Lactose, VP, • Chef reservoirs are reptiles &
tarda • Bacteremia, MR, motility citrate, PD, fresh-water fish
wound infections urease • Infections often involve aquatic
environments
• Pos indole differentiates from
Salmonella
Salmonella Typhoid (enteric) fecer, H2S, MR, motility, Lactose, indole, • Found in poultry
bacteremia, enterocolitis lysine VP, PD, urease, • May be transmitted by reptiles
decarboxylase ONPG • S. typhi has Vi antigen, only trace
(LDC) H2S citrate neg
• Grouped by O antigens (e.g.
A,B,C), serotyped by H antigens
(e.g. 1,2)
Citrobacter Nosocomial infections Gas, H2S, MR, VP, PD, LDC • Lactose variable
freundii citrate, motility, • ONPG & LDC differentiate from
ONPG Salmonella
Klebsiella Pneumonia, UTI, Lactose, gas, VP, H2S, indole, • Encapsulated
pneumoniae septicemia citrate, urease MR, PD, motility, • Colonies usually mucoid
(slow) ornithine • Some strains hydrolyze urea
decarboxylase slowly
(ODC) • K. oxytoca is similar to K.
pneumoniae except indole pos
• Motility & ODC differentiate from
Enterobacter
Enterobacter • Opportunistic & Lactose, gas, VP, H2S, indole, • Colonies may be mucoid
aerogenes & nosocomial citrate, motility, MR, PD • Same IMViC reactions as
cloacae infections ODC Klebsiella
• UTI, RTI, & wound
infections

Page 8 | W e e k 1 3
 WEEK 13: “Enterobacteriaceae”

ORGANISM TSI MacCONKEY HEKTOEN ENTERIC XLD

Escherichia coli A/A, gas Flat, dry pink colony with Yellow Yellow
darker pink halo
Shigella K/A Colorless Green Colorless
Edwardsiella K/A, gas, H2S Colorless Colorless Red, yellow, or colorless
with or without black
centers
Citrobacter A/A or K/A gas, with Colorless at 24 hr Colorless Red, yellow, or colorless
or without H2S May become pink at 48 hr with or without black
centers
Salmonella K/A, gas, H2S Colorless Green Red with black center
Klebsiella A/A, gas Pink, mucoid Yellow Yellow
Enterobacter A/A, gas Pink Yellow Yellow
May be mucoid
Serratia K/A Colorless at first, turning Colorless Yellow or colorless
pink
S. marcescens may have
red pigment at RT
Proteus K/A (mirabilis) A/A Colorless Colorless Yellow or colorless, with
(vulgaris), gas, H2S May swarm or without black centers
Morganella K/A, gas Colorless Colorless Red or colorless
Providencia K/A Colorless Colorless Yellow or colorless
Yersinia yellow/orange Colorless to peach Salmon Yellow or colorless

Table 19-11 Stool Culture Screening for Enteric Pathogens Using Triple Sugar Iron and Lysine-Iron Agar in Combination
TSI REACTIONS
LIA K/A H2S K/AG H2S K/AG K/A A/A H2S A/AG A/A K/K
Reactions
R/A P. vulgaris M. morganii M. morganii P. vulgaris -- Providencia --
P. mirabilis Providencia Providencia P. mirabilis
K/K H2S Salmonella* Salmonella* Salmonella* Salmonella* -- -- -- --
Edwardsiella Edwardsiella
K/K Salmonella -- Hafnia Salmonella* Klebsiella Serratia Pseudomonas
Klebsiella Plesiomonas+ Enterobacter
Serratia Hafnia E. coli
K/A H2S -- Salmonella* -- Serratia -- -- -- --
K/A -- Citrobacter Salmonella Shigella* Citrobacter Aeromonas*+ Aeromonas*+ --
Shigella Yersinia E. coli Yersinia
Aeromonas Aeromonas+ Citrobacter Citrobacter
E. coli E. coli Enterobacter Enterobacter
Enterobacter Enterobacter
Citrobacter

Page 9 | W e e k 1 3