Gram Negative Cocci and

Bacilli
Franzielle Jowe D. Cabrera
GRAM NEGATIVE COCCI
 Neisseria
Gram negative diplococci resembling
__________________
Obligate aerobes
Oxidase and Catalase (+)
Capnophilic
 Neisseria
Culture media
Thayer Martin VCN
Modified TM VCNT
Martin Lewis VCAnT
New York City Agar VCAmT
 ORGANISM ASSOCIATED DISEASE/S
N. gonorrhoeae STD Gonorrhea
Pharyngitis, Proctitis
Ophthalmia Neonatorum
Disseminated Gonococcal
Infection
Fitz Hugh Curtis Syndrome

N. meningitidis Meningococcemia
Wa t e r h o u s e - F r i e d r i c h s e n
Syndrome
Moraxella catarrhalis Localized infections in the
respiratory tract
Neisseria gonorrhoeae
Neisseria meningitidis
 Neisseria
Diagnostic procedures
Superoxyl Test
Carbohydrate Fermentation Test
GLUCOSE MALTOSE SUCROSE LACTOSE

N. gonorrhoeae

N. meningitidis

N. lactamica

N. sicca

M. catarrhalis
GRAM NEGATIVE BACILLI
 Enterobacteriaceae
Gram negative rods
Motile via peritrichous flagella except
_____
Facultative anaerobes
Catalase (+)
Oxidase__________
Nitrate reduction (+)
________ fermenter
 Enterobacteriaceae
Antigenic Structure
K antigen
Capsular antigen
Heat _________
Vi antigen _____________
O antigen
Somatic antigen / cell wall
Heat ____________
Mainly IgM antibodies
H antigen
Located in the flagella
Mainly IgG antibodies
 MacConkey Agar
Selects gram negative bacteria and also
differentiates lactose fermenters (pink-red)
from non-lactose fermenters (colorless)
Bile Salts, Neutral red and
crystal violet inhibit gram
(+) bacteria
Lactose Carbon source
Neutral red brown in pH 6.8
to 8.0 and pink-red at pH less
than 6.8
 Eosin Methylene Blue
Selects gram negative bacteria and also
differentiates lactose fermenters (purple color to
green metallic sheen) from non-lactose fermenters
(colorless).
Eosin and Methylene Blue
inhibit gram (+) bacteria
Lactose Carbon source
 Hektoen Enteric Agar
Selects for stool pathogens by inhibiting normal flora
coliforms and also differentiates lactose fermenters
(orange) from non-lactose fermenters (green)
Bile salts (high amounts)
inhibit gram (+) bacteria and
gram (-) coliforms.
Lactose, sucrose and salicin
Carbon source.
Bromthymol blue indicator
Sodium thiosulfate sulfur
source for H2S production.
Ferric ammonium Citrate - to
detect H2S production.
 Hektoen Enteric (HE) Agar
Selects for stool pathogens by inhibiting normal flora
coliforms and also differentiates lactose fermenters
(yellow or orange) from non-lactose fermenters
(green)

E. coli Salmonella Shigella

 XLD Agar
Selects for stool pathogens by inhibiting normal flora
coliforms and also differentiates xylose fermenters
(yellow) from non-xylose fermenters (red)
Sodium deoxycholate
inhibit gram (+) bacteria, gram
(-) coliforms and swarming of
Proteus
Xylose Carbon source.
Phenol red indicator
Sodium thiosulfate sulfur
source for H2S production.
Ferric ammonium Citrate - to
detect H2S production.
 XLD Agar
Selects for stool pathogens by inhibiting normal flora
coliforms and also differentiates xylose fermenters
(yellow) from non-xylose fermenters (red)

Citrobacter & Proteus

yellow colonies with
black centers
Shigella
red colonies

Xylose Fermenters Non Xylose Fermenter and H2S producer

(yellow colonies) (red colonies with black centers)
Salmonella
 Salmonella-Shigella Agar
Selects for stool pathogens by inhibiting normal flora
coliforms and also differentiates lactose fermenters
(pink-red) from non-lactose fermenters (clear)
Bile Salts and Briliant green
agar inhibit gram (+) bacteria
gram (-) coliforms
Lactose Carbon source.
Neutral red indicator
Sodium thiosulfate sulfur
source for H2S production.
Ferric ammonium Citrate - to
detect H2S production.
Salmonella-Shigella Agar

A. K. pneumoniae
B. E. coli
C. Salmonella
D. Proteus
E. Pseudomonas
aeroginusa
 Cefsulodin-irgasan-novobiocin
Also known as Yersinia Selective Agar
Selective media for Yersinia species
 Enterobacteriaceae
TSI
Sugars:
pH indicator:
H2S indicator:
LIA
pH indicator:
H2S indicator:
Indole
Enzyme detected:
Medium:
Reagent:
Positive result:
 TSI Results

Aerobic slant
portion

Anaerobic
deep portion

Alcaligenes freundii

Pg. 245
LIA Results

Klebsiella

aerogenes
 MRVP
Methyl Red
Mixed acid fermentation:
Reagent:
Positive result:
Voges Proskauer
Detects:
Reagents:
Positive Result:
Citrate
Enzyme present:
Culture media:
pH indicator:
Positive result:
 Malonate
Differentiates Salmonella from Shigella by using
malonate as sole source of carbon

Urease
Culture media:
pH indicator
Positive result:
Rapid urease producers:
Slow urease producers:
Urease Test results
 ONPG
Enzyme detected:
Substrate:
Positive Result:

Decarboxylase
Enzyme removes _________ producing alkaline
products
Culture media:
pH indicator:
 Deaminase
Enzyme that removes _______ resulting in an
acidic product
Reagent:
Positive result for Phenylalanine:
LACTOSE FERMENTERS
 Enterobacteriaceae
ESCHERICHIA
UTI 90%
Neonatal meningitis
Nosocomial, bacteremia and Diarrheal
diseases
Green metallic sheen on EMB
TSI: A/A with Gas IMViC: _________
MUG (+) except E. coli O157:H7
 Pathogenic E. coli
ETEC ______________
LT and ST
LT similar to _________ toxin
Montezumas revenge, travellers diarrhea, watery
stool

EIEC ______________
Shigella like infection (dysentery)
Stool with RBC, Neutrophils and mucus
 EPEC _____________
Non invasive, no toxin
Watery diarrhea with mucus, no blood

EHEC _____________
Verotoxic E. coli (E. coli O157:H7)
Most severe manifestation: ________
SMAC: ___________

EAEC ______________
Adheres to host cells
Watery diarrhea
E. coli - Microscopic Exam
E. coli on EMB Flagellated E. coli
 Enterobacteriaceae
KLEBSIELLA
Mucoid colonies, large polysaccharide capsule
IMViC: - - + + TSI: A/A with gas
_______________ Friedlanders bacillus,
encapsulated and tends to string
K. oxytoca indole positive

ENTEROBACTER
UTI, Respiratory Tract and wound infections
______________ - Most predominant isolate
Encapsulated Klebsiella pneumoniae mucoid colonies
MacConkey Agar
E. Coli and Enterobacter aerogenes E. Coli and Klebsiella pneumoniae
MacConkey Agar MacConkey Agar
Eosin Methylene Blue Greenish Metallic Sheen of
E. coli in EMB
LATE LACTOSE FERMENTER
 Enterobacteriaceae
ARIZONA
Related to Salmonella
____________ lactose fermenter

CITROBACTER
Resembles Salmonella except _________ positive and
________ negative
__________ - causes neonatal meningitis
__________ - UTI, pneumonia, endocarditis

SERRATIA
______________ - produces red pigment
(____________)
Related to nosocomial infections
 Enterobacteriaceae
YERSINIA
Y. pestis
Bipolar stain on ___________
_____________ appearance
_____________ pattern on broth
Causative agent of ____________
Vector _____________
3 types
Y. enterocolitica
Motile at _______ but not at ________
__________ colony on CIN
Causes _______________
Safety pin appearance Yersinia pestis
 In Europe, black rats
and Norway rats.
Widely blamed for the
European plague
epidemics in the Middle
Ages.

Humans can contract bubonic

plague when bitten by a flea that
is carrying the plague bacterium
In the American Southwest
(prairie dogs)
 Bubonic plague occurs within a week of being
bitten by an infected flea. Multiplication of the
bacteria produces the characteristic
"bubo" (swollen, painful lymph node).
Bacteremia follows, causing death in about 75%
of those affected.
Swollen lymph node
Pneumonic plague occurs under crowded
conditions when contaminated respiratory
droplets expelled by infected persons are
directly inhaled by another person. This form is
characterized by a shorter incubation period
and greater mortality (90%).
NON-LACTOSE FERMENTERS
 Enterobacteriaceae
SALMONELLA
Black colonies on SSA, BSA
Ingestion of milk, dairy products, shellfish, eggs,
meat
Species:
__________ typhoid fever, meningitis, osteomyelitis
S. paratyphi A and B paratyphoid fever
S. paratyphi C ( S. cholera suis ) septicemia
S. enteritidis gastroenteritis
___________ - tube dilution agglutination test for
detection of Salmonella infection
 Enterobacteriaceae
SHIGELLA
Non-motile, colorless colonies on SSA
Intestinal pathogen causes ____________
O Ag ONPG CATALASE MANNITOL
A

B

C

D

 Shigella
Shigellosis or Bacillary Dysentery
watery diarrhea
bloody stool with WBCs & mucus
fecal-oral route (human only known
reservoir)
self-limiting but highly communicable bec. of
low infective dose (200 bacilli)
Gay bowel syndrome (S. flexneri)
 Shigella
Reiter's syndrome is a late complication of
S. flexneri infection. (3%)
pains in their joints, irritation of the eyes,
and painful urination
can lead to chronic arthritis
 Enterobacteriaceae
PROTEUS, PROVIDENCIA, MORGANELLA
LDA positive
Urease positive
R/A on TSI
Proteus
Swarming motility
P. vulgaris indole ________
P. mirabilis indole ________
 Proteus
Swarming phenomenon
Proteus mirabilis
 Enterobacteriaceae
EDWARDSIELLA
Similar to __________ on IMViC
TSI similar to ___________
__________ - most common human species
 PSEUDOMONAS AERUGINOSA
Motile and rod shaped, obligate aerobe
Sweet or grape-like, corn-tortilla like odor due
to _____________
Grows well at ________ deg C
Oxidase positive
TSI reaction ____________
Produces blue-green pigment
Blue
Yellow
 PSEUDOMONAS AERUGINOSA
Wound infections causing blue-green pus
Meningitis, pneuomonia, sepsis
Mild otitis media/externa -
______________
Skin lesions following bacteremia -
________
 BURKHOLDERIA
BURKHOLDERIA MALLEI
Causative agent of _____________ , disease of the
horses

BURKHOLDERIA PSEUDOMALLEI
Causative agent of _____________ , glanders like
disease
Also known as ______________ bacillus

BURKHOLDERIA CEPACIA
Commonly isolated on patients with _______________
 Vibrio
Facultative anaerobe
Curved, comma shaped, motile via _____
flagella
Oxidase _________
0129 ___________
Halophilic except _________________________
Sucrose fermenter except
___________________
Alkaline loving: APW and TCBS
 Vibrio
Species
V. cholerae
2 strains ________________
Rice watery stool
String test (+)
Reagent: _______________
3 serogroups: _____________________
2 biovars: ____________________
 CLASSICAL EL TOR
RBC hemolysis

VP

Polymxyin B

RBC agglutination

 V. parahaemolyticus
2nd most common sp implicated in
gastroenteritis
1 cause of summer diarrhea in Japan
Associated with consumption of improperly
handled seafood (oysters, clams, crabs,
lobster, scallops, sardines and shrimp)
Kanagawa phenomenon
_______________
 AEROMONAS HYDROPHILIA
Oxidase ____________
_____________ hemolytic

PLESIOMONAS SHIGELLOIDES
Oxidase ____________
_____________ hemolytic

STENOTROPHOMONAS MALTOPHILIA
Oxidizes _________ faster than glucose
Lavender green colony
Oxidase _______
 ACINETOBACTER
Oxidase _____ , non-motile
UTI, wound infection, diarrhea
_____________ - oxidixer
_____________ - non-utilizer

ALCALIGENES FAECALIS
Oxidase and Catalase _________
Asaccharolytic
Apple like odor
 Haemophilus
Non motile
Facultative anaerobes
Fastidious _________
Staph streak
 Haemophilus
H. influenzae
AKA ____________________
Six serotypes; most commonly encountered __________
Meningitis among children <5 years old
Causes epiglottitis
H. aegyptius
AKA ______
Causative agent of __________
H. ducreyi
Causative agent of ___________
Short bacilli in __________________ arrangement
 Haemophilus
D-ALA X FACTOR V FACTOR
(Porphyrin test)
H. influenzae

H .
parainfluenzae

H .
parahemolyticus

H. aegyptius

H. ducreyi

 CAMPYLOBACTER
C. jejuni and C. coli (associated with
gastritis and diarrhea)
Microaerophilic and Capnophilic
Optimum growth temperature
___________
Motility __________
Special culture media ___________
Microscopic _____________
 HELICOBACTER
Associated with __________
Found in human stomach
Relationship between H. pylori and ulcers
 BORDETELLA
Agent of whooping cough ______________
3 stages
Catarrhal
Paroxysmal
Convalescent
Pertussis like syndrome ________________
Only motile species _________________
Special culture media
___________________
 BRUCELLA
Causes ___________________
Morning NO FEVER
Night WITH FEVER
Infection acquired through ingestion of
animal products and via direct contact
Special culture media
__________________
 GROWTH IN THIONINE GROWTH IN BASIC
FUCHSIN
B. abortus

B. melitensis

B. suis

B. canis

 LEGIONELLA
Legionnaires disease _______________
Pontiac fever
Found in both natural and artificial water
sources
May be present inside free living amoebas
Special culture media _______________
 FRANCISELLA TULARENSIS
Faintly staining, gram negative
coccobacilli
Causes _____________
Special growth requirement
_____________
Infection via inhalation and via vectors
______
Special culture media _______________
 PASTEURELLA
____________ agent of pasteurellosis
Present in the oral cavity of domestic
animals (cat and dogs)
Bipolar staining, safety pin appearance,
with capsule
______________ are the most common
manifestation
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