Gram Negative Bacteria

MEDICALLY IMPORTANT BACTERIA

Gram Negative Cocci
Neiserria
Moraxella
Neisseria

 Important characteristics:
 Gram negative diplococcic resembling coffee beans or
kidney beans except N. elongata (rod-shaped).
 Obligate aerobes
 Capnophilic –requires 5-10% CO2
 Nonmotile
 Catalase positive except N. elongata
 Oxidase positive
Neisseria spp.

Neisseria gonorrhoeae Neisseria meningitidis

 Gonorrhea/ CLAP, STD  Thrombocytopenia
 Gonococcal Arthritis  Disseminated intravascular
 Ophthalmia neonatorium coagulation
 Shock
 Waterhouse-Friderichsen
syndrome ( hemorrhage in adrenal
gland)
 Moraxella catarhalis
Other Neisseria (Branhamella catarhalis/ Neisseria catarhalis)

 Rarely involved in human infection.  Most infection are localized to sites

associated with the respiratory
 Bacteremia
tract that include sinusitis and
 Endocarditis pneumonia.
 Meningitis  Nonmotile
 Aerobic
 Catalse +
 Oxidase +
 Nonfermenter of CHO
 DNAse +
Presumptive test

 Superoxol Test
 Reagent : 30% H2O2
 Positive result: Bubbles (purple); Neisseria gonorrhoeae
 Carbohydrate Fermentation of Neisseria spp.
Neisseria spp GLUCOSE MALTOSE SUCROSE LACTOSE

Moraxella catarhalis Negative Negative Negative Negative

(DNAse test = Positive)
Neisseria gonorrhoeae Positive Negative Negative Negative

Neisseria meningitidis Positive Positive Negative Negative

Neisseria subflava Positive Positive Positive Negative

Neisseria lactamica Positive Positive Negative Positive

(ONPG test = Positive)
Cultivation

Chocolate Agar Plate Modified Thayer Martin

Martin Lewis Thayer Martin
New York City Agar
 Composition of Culture media used for
Neisseria spp.
Thayer Martin Modified Martin Lewis New Your City Agar
Thayer Martin
Vancomycin Vancomycin Vancomycin Vancomycin
Colistin Colistin Colistin Colistin
Nystatin Nystatin Trimethoprim lactate Trimethoprim lactate
Trimethoprim lactate Anisomycin Ampothericin B
Gram Negative Bacilli
Enterobacteriacea Streptobacillus
Pseudomonas Capnocytophaga
Burkholderia Eikenella
Vibrio Calymmatobacterium
Haemophilus Chromobacterium
Campylobacter Cardiobacterium
Helicobacter
Bordetella
Brucella
Legionella
Pasteurella
Francisella
Gardnerella
Enterobacteriaceae
 Gram negative rods
 Motile with peritrichous flagella except Klebsiella and
Shigella
 Facultative anaerobes
 Ferment glucose
 Often with gas production
 Catalase positive
 Oxidase negative
 Reduce nitrate to nitrite except for Erwinia and Pantoea
agglomerans
Antigenic Structure

 K antigen
 Capsular antigen
 Heat-labile
 Some Salmonella have capsular (K) antigen, referred to
as Vi (virulence antigen)
 O antigen
 Somatic antigen
 Heat-stable
 Antibodies to O antigens are predominantly IgM
 H antigen
 Located in the flagella
 Denatured or removed by heat or alcohol
 H antigens agglutinate with anti-H antibodies, mainly IgE
Laboratory Diagnosis

Growth on MacConkey Agar Eosin-Methylene Blue Agar

 Selective/differential for Gram  Selective/differential for Gram
negative Enteric bacilli
negative Enteric bacilli
 CHO: Lactose
 Inhibitor for Gram +: Crystal violet  LF: pink to purple colonies
and Bile salt  NLF: colorless colonies
 CHO: lactose  Characteristics colonies on EMB
 Escherichia coli: pink to purple
 pH indicator: neutral red colonies with green metallic sheen
 LF: pink colonies  Klebsiella: pink mucoid colonies
 NLF: colorless colonies  Enterobacter: pink colonies with a
dark center (fish eye colonies)
Hektoen-Enteric Agar Salmonella-Shigella Agar
 Selective/differential for Gram negative  Selective/differential for Salmonella
Enteric bacilli
and Shigella
 Inhibitor for Gram +: bile salt
 CHO: lactose, sucrose and salicin  CHO: Lactose
 pH indicator: Bromthymol blue  pH inhibitor: neutral red
 H2S: ferric ammonium citrate
 H2S indicator: ferric citrate
 LF, H2S negative: yellow without black center
 LF, H2S positive: yellow with black center  Salmonella: NLF, H2S + ; colorless
 NLF, H2S negative: green colonies without colonies with black center
black center
 NLF, H2S positive: green colonies with black
 Shigella: NLF, H2S - ; colorless
center colonies without black center
MAC EMB
HE SS
Triple Sugar Iron Agar Lysine Iron Agar
 Sugars: 10 parts lactose, 10 parts  Detects lysine deamination and
sucrose and 1 part glucose decarboxylation
 pH indicator: phenol red  pH indicator: bromcresol purple
 H2S indicator: sodium thiosulfate is  H2S indicator: ferric ammonium
reduced to SO2 and H2S, and H2S citrate
reacts with ferrous sulfate to
produce black ferric sulfide.
Triple Sugar Iron Agar

 Considered an initial step in the identification of

Enterobacteriaceae
 Composition:
 Protein source
 CHO: lactose (10) 1%
 sucrose (10) 1%
 glucose (1) 0.1%
 pH indicator: phenol red
 H2S indicator: ferrous sulfate
 TSI

Butt/Slant Gas, H2S

Butt: Glucose fermenter
Slant: Lactose and Sucrose fermenter
Gas: Cracks (All enterobacter are AEROGENIC
except SHIGELLA
H2S: Blackening of the medium
Sulfur source: Sodium thiosulfate

H2S + organism:
Salmonella
Proteus
Arizona
Citrobacter
Edwardsiella
TSI reactions
TSI reaction Typical Organism

A/A G,H2S- Yellow = LF/SF Gas + Escherichia

Yellow = GF H2S - Klebsiella
Enterobacter

A/A G, H2S+ Yellow = LF/SF Gas + Salmonella

Yellow = GF H2S + Proteus

K/A H2S- Red = NLF/NSF Gas – Shigella

Yellow = GF H2S -

K/K H2S- Red = NLF/NSF Gas - Pseudomonas

Red = NGF H2S - Alcaligenes
Lysine Iron Agar

 Can be used t determine the ability of the organism to

deaminate lysine, decarboxylate lysine and produce H2S
 Composition:
 Small amount of protein
 Glucose. Lysine
 pH indicator: Bromcresol purple
 H2S indicator: Ferric ammonium citrate
LIA

 Uninoculated LIA: PURPLE

Slant: Lysine Deamination (PPM)
(+): RED
(-): PURPLE

Butt: Lysine Decarboxylation

𝐺𝑙𝑢𝑐𝑜𝑠𝑒 𝐿𝐷𝐶
Purple 𝐹𝑒𝑟𝑚𝑒𝑛𝑡 Yellow 𝐴𝑙𝑘𝑎𝑙𝑖𝑛𝑒 𝑝𝑟𝑜𝑑𝑢𝑐𝑡𝑖𝑜𝑛 (+) PURPLE; (K)
(𝐶𝐴𝐷𝑉𝐸𝑅𝐼𝑁𝐸)
(-) YELLOW

H2S (+): Blackening of the medium

SACEd

* LIA is not sensitive to H2S production of Proteus.

 LIA reaction
LIA reaction Typical Organism

K/K H2S+ Purple H2S + Salmonella

Purple

Lysine deaminase= negative

Lysine decarboxylation= positive
K/A H2S - Purple H2S - Shigella
Yellow

Lysine deaminase= negative

Lysine decarboxylation= negative
R/A H2S- Red H2S - Proteus
Yellow Providencia
Morganella
Lysine deaminase= positive
Lysine decarboxylation= negative
Indole test

 Detects: Tryptophanase
 Medium: Tryptophan broth, SIM (sulfindole motility
medium)
 Indicator: Kovac’s or Ehrlich’s reagent (P-DAB)
 Positive result: formation of red ring
𝑡𝑟𝑦𝑝𝑡𝑜𝑝ℎ𝑎𝑛𝑎𝑠𝑒
 Tryptophan indole + indicator
MRVP test
 Medium: Methyl Red-Voges Proskauer Broth or Peptone Glucose Broth
 A. Methyl Red
 Reagent: Methyl red (pH indicator)
 pH: <4.5
 Organism that produce enough acid will overcome the neutralizing effect of
the buffer
 Positive result: RED
 Mixed Acid production pathway:
 Lactic acid
 Acetic acid
 Formic acid
 Succinic acid
MVRP test

 B. Voges-Proskauer
 Detects: Acetoin (acetylmethyl carbinol)
 Positive result: RED
 Butyrene Glycol Pathway:
 Acetoin (acetylmethyl carbinol)

Most Enterobactericeae, give opposite MR and VP

reactions
Citrate Utilization test

 Determine of an organism can utilize citrate as sole

source of carbon
 Medium: Simmon’s Citrate Agar
 pH indicator: Bromthymol blue
 Positive result: Growth and Intense blue
Malonate Utilization

 Determine if an organism can utilize malonate as a sole

source of carbon
 Positive result: BLUE
 Negative result: Yellow or Green
Urease Test

 Detects UREASE
 Medium: Christensen’s urea Agar or Shewart’s Urea
Broth
 pH indicator: phenol red
 Positive result: Red or Hot Pink (Crease Pink)
 Rapid Urease production (w/in 2-4 hours): PPM
 (Proteus, Providencia, Morganella)
 Slow Urease production (after 4 hours): CKEYS
 (Citobacter, Klebsiella, Enterobacter, Yersinia, Serratia)
ONPG test

 Identifies late LACTOSE FERMENTER

 Rapid test to detect: β-galactosidase
 Substrate: ONPG (ortho-nitrophenyl-beta-D-
galactopyranoside)
 Positive: Yellow color
β−galactosidase
 ONPG Galactose + ortho-nitrophenyl
Decarboxylase Test

 Enzyme that removes the carboxyl group (COOH) from

the amino acid
 Uses decarboxylase basal medium with indicator
(bromcresol purple) + sugar (glucose) + amino acid
(lysine, ornithine, arginine) + control tube with no amino
acid
 Moller’s medium better
 Positive result: Purple to yellow to PURPLE
Phenylalanine deaminase (PDA)

 Enzyme that removes amino group (NH2) from an amino

acid
 Phenylalanine agar
 Overnight culture + 10% Ferric chloride
 Positive result: Green slant and fluid (PPM)
Gelatin liquefaction

 Determine if an organism can breakdown gelatin into

Amino Acids
 Positive result: LIQUEFACTION
 Serratia
 Proteus
Escherichia

 IMViC: ++--
 TSI: A/A +-
 UTI –90%
 Sepsis
 Meningitis – (also Group B Strep) causes meningitis in
infants
 Diarrheal disease
Enterotoxigenic E. coli (ETEC) Enteroinvasive E. coli (EIEC)
 Produces heat-labile (LT) and/or heat-  Invades the intestinal epithelium
stable (ST) enterotoxins; genes of causing SHIGELLA-LIKE
both toxin reside on a plasmid. INFECTION.
 LT are closely related in structure and  Dysentery (necrosis, ulceration and
function to CHOLERA TOXIN; ST
inflammation of large bowel);
result in net intestinal fluid secretion
usually in young children living in
by stimulating guanylate cyclase.
areas of poor sanitation
 “Montezuma’s revenge” or “Turista”,
traveler’s and childhood diarrhea  Stool with RBC, Neutrophils and
chratacterized by profuse watery stool. mucus
 Enterohemorrhagic E. coli (EHEC)/
Enteropathogenic E. coli (EPEC) Verotoxic E. coli (VTEC)
 Non-invasive, produces no toxin  Produce verotoxin – named for its
cytotoxic effect on Vero cells, a line of
 Nosocomial infection seen in African Green Monkey kidney cells, a
cytotoxin that resembles that of
newborn and infants SHIGELLA DYSENTERIAE.
 Watery diarrhea with mucus but no  HUS – most severe manifestation of
EHEC
blood  Most frequently associated with certain
serotypes such as Escherichia coli
O157:H7
 MacConkey agar in which is SORBITOL
has been substituted for lactose- only
strain that is sorbitol negative- colorless
colonies
Enteroaggregative E.coli (EAEC)
 Probably involves binding by pili,
ST-ike, and hemolysin-like toxins;
actual pathogenic mechanism not
known
 Water diarrhea
Klebsiella

 IMViC: --++
 TSI: A/A +-
 Nonmotile
 Exhibit mucoid growth, large polysaccharide capsule
 K. pneumoniae- Friedlander’s bacillus, encapsulated and
appears as mucoid colonies that tend to string.
 K. ozanae- purulent sinus infection
 K. rhinoscleromatis- granuloma of the nose and oropharynx
 K. oxytoca- Indole positive
Enterobacter

 IMViC: --++
 TSI: A/A +-
 Motile
 Oppurtunistic infections: UTI, RT, and wound infections
 Most predominant isolate is: Enterobacter cloacae
 Produces yellow pigmentation that intensifies at 25˚C
(Enterobacter sakazakii)
 Klebsiella VS Enterobacter
LDC ODC ADH
(Lysine = Cadaverine) (Ornithine = Putrescine) (Arginine = Citruline
to Ornihine)
Klebsiella pneumoniae Positive Negative Negative

Klebsiella oxytoca Positive Negative Negative

Enterobacter cloacae Positive Positive Negative

Enterobacter aerogenes Negative Positive Positive

Pantoea agglomerans Negative Negative Negative

Serratia

 DNAse = +
 Gelatinase = +
 Lipase = +
 Serratia marcescens and Serratia rubiudea produce red
pigment (Prodigiosin – not diffusible pigment)
 Serratia odorifera produces a rancid-odor potato-like odor
 Common opportunistis pathogen in hospitalized patients
Hafnia

 Hafnia alvei – formerly known as Enterobacter alvei

 To differentiate from other Enterobacter
 Slow fermenter of Lactose
 Citrate negative
 Salmonella
 Sources of infection: water, milk and diary products, shellfish (from
contaminated water), dried or frozen eggs, meat and meat
products and household pets
 Cause enteritis, systemic infection and enteric fever (typhoid fever)
 Cause enteric fever:
 Salmonella paratyphi A
 Salmonella paratyphi B
 Salmonella cholerasuis - bacteremia
 Salmonella typhi (most important cause)
 Endocolitis/gastroenteritis – most common manifestation of
Salmonella infection – Salmonella typhimurium (most common)
Widal’s test

 Tube dilution agglutination test

 At least two serum specimens, obtained at intervals of 7-10
days, are needed to prove a rise in antibody titer
 Serial dilutions of unknown serum are tested against antigens
from representative Salmonellae
 High or rising titer of O ≥1:160 suggest that active infection is
present
 High titer of H ≥1:160 suggests past infection or immunization
 High titer antibody to Vi antigen occurs in some CARRIERS
(Mary Mallon).
 Shigella

 Mostly NEGATIVE reactions

 Natural habitat of shigellae is limited to the intestinal
tracts of humans and other primates, where they produce
BACILLARY DYSENTERY.
Present Group type Catalase ONPG Mannitol
Designation
Shigella dysenteriae A Negative Negative Negative

Shigella flexneri B Positive Negative Positive

Shigella boydii C Positive Negative Positive

Shigella sonnei D Positive Positive (late LF) Positive

Citrobacter

 Causes UTI and sepsis

 Organisms resemble Salmonella but are ONPG positive
and LDC negative
PPM

 Nonfermenter for lactose

 Motile
 MR +
 Lysine deamination +
 LIA: R/A
Proteus

 Produce infections in humans only when they leave the

intestinal tract
 UTI, bacteremia, pneumonia, nosocomial infections
 P. mirabilis and many strains of P. vulgaris enhibit swarming
motility
 Certain strains share specific polysaccharides with some
rickettsia and are agglutinated by sera from patients with
rickettsial diseases:
 A. Proteus mirabilis – OX-K
 B. Proteus vulagris – OX-19 and OX-2
 Proteus mirabilis Proteus vulgaris

LOA (-, -, -) (-, +, -)

Indole (+) (-)

Chloramphenicol Resistant Sensitive

Providencia

 Normal intestinal flora

 UTI and occasionally other infections and are often
resistant to antimicrobial therapy

Providencia rettgeri Providencia stuartii

Urease Positive Negative
 Morganella Providencia
(++--) (++-+)
H2S (-) (-)

Citrate (-) (+)

Gas (+) (-)

Edwardsiella

 IMViC: ++--
 TSI: A/A +-
 Isolated from the environment and many cold-blooded
and warm blooded animals including reptiles, freshwater
and aquarium fish, frogs and turtles.
 Edwardsiella tarda- most human species
 To differentiate from Escherichia coli
 A. non-fermenter of Lactose
 B. H2S= +
Yersinia

 Motile at room temperature (25˚C) but non-motile at 37˚C

 Yersinia pestis
 Plague (black death) – infection of wild rodents transmitted
from ione rodent to another and occasionally by the bite of
fleas
 Common vector: Xenopsylla cheopis – rat flea
 Produced pandemics of black death with million of fatalities
 Broth cultures exhibit a characteristic “Stalactite pattern” in
which clumps of cells adhere to one side of the tube
 Inclusion: BIPOLAR BODIES
 Stain: WAYSON’S Stain = SAFTEY PIN APPERANCE
 Yersinia enterocolitica
 Enterocolitis- characterized by fever, diarrhea and
abdominal pain
 Bacteremia
 Cultured on: CIN(cefsulodin-Irgasan-Novobiocin)
 Bull’s eye colonies
Pseudomonas aeruginosa

 Obligate aerobe
 Motile and rod shape
 Sweet or grape-like or corn-taco like odor resulting from the
production of 2-aminoacetophenone
 Grows well at 37˚C, its growth at 42˚C will differentiate it from
other Pseudomonas spp.
 Oxidase positive
 Does not ferment CHO: TSI = K/K
 Produces blue-green pigment: diffuse in medium
 * Pyocyanin = Blue
 * Pyoverdin = Green
 Produces infection of wound and burns, giving rise to
blue-green pus
 Menigitis, UTI, Pneumonia, Sepsi, Mild otitis
esternia/media (Swimmer’s Ears)
 Skin lesion- Ecthema Gangrenosum
 Resistant to a number of disinfectants and has been
responsible for serious NOSOCOMIAL INFECTIONS
Burkholderia mallei

 Causes GLANDER’S a disease of horses and similar

animals transmissible to humans
 Horses – pulmonary involvement
 Human – fatal, begin as ulcer of skin and mucous
membrane followed by lymphangitis and sepsis
Burkholderia pseudomallei

 Whitmore’s bacillus
 Causes Malioidosis, an epidemic glander’s like disease of
animals and humans
 Vietnamese time bomb disease
Vibrio

 Facultative anaerobes
 Monotrichous
 Oxidase positive = (+) Dark purple
 Halophilic excepy Vibrio mimicus and Vibrio cholerae
Vibrio cholerae

 O1 and O139- pandemics spread of disease

 Characteristics rice watery stool
 Cultured on APW and TCBS
 String test
 Reagent: 0.5% Na desoxycholate
 Serogoups:
 Inaba - Philippines
 Ogawa - India
 Hikojima - Japan
 Biotypes (Biovars)

Classical El tor
(Past Pandemics) (Recent Pandemics)
Red cell hemolysis Negative Positive
VP Negative Positive
Polymyxin B Sensitive Resistant
Agglutination with chicken RNCs Negative Positive
Culture

Alkaline Peptone Water Thiosulfate Citrate Bile Salt Sucrose Agar

 0.5 to 1 % NaCl  CHO: Sucrose

 pH indicator: Bromthymol blue
 SF: yellow colonies (Vibrio cholera)
 NSF: green colonies (Vibrio
parahemolyticus)
Alkaline Peptone Water TCBS
Other Vibrio spp

Vibrio alginolyticus Vibrio parahemolyticus

 Blood, eye, and ear infection  Clinical significance:
 Clinical significance: wound and gastroenteritis, usually associated
ear infections associated with with contaminated seafood
marine environment  Halophilic
 Halophilic  Non-sucrose fermenter
 Sucrose fermenter
Vibrio mimicus Vibrio vulnificus
 Clinical significance: gastroenteritis  Clinical significance: septicemia
and ear infections associated with and wound infections involving
marine environment marine environment
 Nonhaophilic  Halophilic
 Non-sucrose fermenter
Haemophilus

 Non-motile
 Non-sporeforming
 Facultative anaerobes
 Most spp. are oxidase and catalase positive
 Preferred incubation: 35-37˚C with 5-10% CO2
Haemophilus spp.

Haemophilus influenzae Haemophilus aegypticus

 Formerly known as PFEIFFER’S  Formerly known as KOCH-
BACILLI WEEK’S BACILLI
 Six serotypes (a,b,c,d,e,and f)  Closely resembles Haemophilus
 Most frequently encountered serotype influenzae biotype III
in infection (𝐻𝑖 𝑏 )
 Causes PINK EYE Conjuctivitis
 Encapsulated strains are pathogenic
 Main cause of meningitis in children
<5 years old
 Associated with respiratory conditions
including epiglottitis
Haemophilus ducreyi
 Infective agent of CHANCROID/
SOFT CHANCRE, venereal
disease characterized by painful
ulcers in the genitalia
 Direct examination short bacilli in a
“school of fish” arrangement
 Smallest pathogenic bacilli
 Culture for Haemophilus spp.
Species X factor V factor Beta hemolysis D-ALA

H. influenzae + + - +

H. parainfluenzae - + - +

H. hemolyticus + + -
+
H. parahemolyticus - + +
+
H. aegypticus - -
+ +
H. aphrophilus - +
- -
H. paraphrophilus - + -
+
H. ducreyi - - -
+
Campylobacter
 S-shaped or seagull wing appearance
 5% O2, 10% CO2, 85% N2
 C. jejuni, C. coli (associated with gastritis and diarrhea)
 Microaerophilic and capnophilic, motile gram negative rods that are associated with gastritis and
diarrhea
 Optimum temperature for growth: 42-43˚C
 Motility: by means of single polar flagella= “DARTING MOTILITY”
 Culture medium: SKIRROW’S agar, Campy’s Blood Agar Plate
 Human pathogen:
 C. coli, C. jejuni = gastritis and diarrhea
 Animal pathogen:
 C. fetus = abortion on animals
 = does not exhibit growth at 42-43˚C
Helicobacter
 35-37˚C
 Associated with PEPTIC ULCER
 The natural habitat is the human stomach, where the organism is found in
the mucus secreting cells
 Current evidence may suggest a role between H. pylori and peptic and
duodenal ulcers
 Histology staining and culture of biopsies obtained from the stomach or
duodenum are recommended for the identification
 S-shaped or Seagull wing appearance
 Microaerophillic organism: 5% O2, 10% CO2, 85% N2
 Motile by means of 4-6 polar flagella
 Urease positive
Bordetella pertussis

 Agent of Whooping cough

 Non-motile except Bordetella bronchseptica
 Three stages of pertussis (whooping cough)
 * Catarrhal: general flulike symptoms
 * Paroxysmal: repetitive coughing episodes
 * Convalescent: recovery phase
 Ideal specimen: Nasopharyngeal swab
Culture

Bordet Gengou Medium Regan-Lowe (Charcoal-Cephalexin)

 Potato  Horse blood
 Blood  Charcoal
 Glycerol  Antibiotics (Cephalexin and
Amphotericin B)
 Preferred for isolation of Bordetella
pertussis
 Note: Both media have a
MERCURY DROPLET COLONIES
of Bordetella pertussis
Bordet Gengou Medium Regan-Lowe (Charcoal-Cephalexin)
Other Bordetella spp.

Bordetella parapertussis Bordetella bronchiseptica

 Pertussis-like syndrome  Inhabits respiratory tract of canines
(kennel’s cough), infrequently
responsible for chronic respiratory
tract infections in human
 Urease Motility Nitate Oxidase
B. pertussis
- - - +
B. parapertussis
+ - - -
B. bronchiseptica
+ + + +
Brucella

 Agents of brucellosis, undulant fever, Malta fever

 Small, nonmotile, aerobic
 Organisms are normal flora of the urinary tract and
gastrointestinal tracts of sheep/goat (B. melitensis),
cattles (B. abortus), pigs (B. suis), and dogd (B. canis)
 Humans acquire infections through the ingestion of
contaminated animal products, including meats and
MILK, farmers can directly acquire infections through
direct animal contract.
 CO2 H2S Growth in the presence of

Thionine Basic Fuchsin

B. abortus + + Inhibit Grow

B. melitensis - - Grow Grow

B. suis - - Grow Inhibit

B. canis - - Grow Inhibit

Culture for Brucella: Castañeda-biphasic medium bottles (Agar and Broth

components)
Incuabte for: 3-4 weeks
Specimen: Blood or Bone Marrow
Legionella

 Legionella pneumoniae: Legionnaire’s disease, Pontiac fever

 Legionella micdadei: Pittsburgh pneumonia
 Legionella bosemanni: Wiga’s agent of pneumonia
 Naturally found in both natural and artificial water sources; the
bacteria have been found in ponds, creeks and streams, wet
soil, water cooling towers for air conditioning and heating
systems, shower heads and plumbing systems
 Culture medium: BCYE (Buffered Charcoal Yeast Extract or
Feeley-Gorman medium
 Specimen: expectorated sputum and bronchial wash
 Stain: DIETERLE’S Stain (Brown to Black stain)
Pasteurella

 Pasteurella multocida (agent of pasteurellosis) wound infection

from animal bite/stratch
 The organism is carried in the oral cavity and respiratory and
gastrointestinal tracts of cats and dogs
 Agent of SHIPPING FEVER in cattles, a hemorrhagic septicemia
 Humans may require the infection following contact with domestic
animals that harbor the bacterium; most frequently the route of
infection is from the bite or scratch of an infected animal, usually a
cat
 Wound infections are the most common type of Pasteurella
infection
 Cats scratch disease
Francisella

 Requires cysteine and cystine

 Faitnly staining, non-motile and obligate aerobic
 Tularemia is a disease of the rodents, primarily rabbits
 Individuals who handle the infected animal, acquire this
zoonosis through direct contact with blood or through
animal bite or scratch
 Bacterium can also be inhaled or acquired indirectly
through insect vectors primarily ticks
 Culture medium: Blood-glucose-cystine medium
Gardnerella

 Garnerella vaginalis (associated with bacterial vaginosis)

 Previously known as Haemophilus vaginalis and
Corynebacterium vaginalis
Bacterial vaginosis

 Is characterized by foul-smelling, grayish, vaginal

discharge
 Amsel and Nuget scoring systems are used to diagnose
BV
 Cytology/Paps: clue cells –epithelial cells studded by
bacteria
 Whiff of Shiff test: vaginal + 10% KOH= (+) Fishy Amine-
like odor
 Organism can be isolated on: Human Blood Bilayer
Tween medium
Streptobacillus moniliformis

 Normal flora of rodents (RATS)

 Agent of:
 1. Rat Bite Fever: Animal bite/scratch= SM/SM (Spirillum
minor,major)
 2. Haverhill Fever: Ingestion of Milk (contaminated)
Capnocytophaga

 Capnophilic
 Fusiform or filamentous bacilli
 Characteristic motility: GLIDING motility
Calymmatobacteria granulomatis

 Agent of DONOVANOSIS= granuloma inguinale

 Encapsulated, pleomorphic bacilli, usually observed in
vacuoles in large mononuclear cells
 Groups of organisms are seen within mononuclear cells;
this pathognomonic entity is known as DONOVAN
BODIES
 Closely related to Klebsiella (Klebsiella granulomatis)
 Based on DNA-DNA hydbridization, it has been recently
proposed that the name of this organism be changed to
Klebsiella granulomatis
Eikenella corrodens Cardiobacterium hominis
 Fastidous, capnophilic rod, part of  Normally found in the upper
the gingival and bowel flora respiratory tract and is isolated as
 Corroding bacterium a rate cause of endocarditis
 Characteristic in agar: PITS or
CORRODES AGAR
 Produces BLEACH-LIKE odor
Chryseobacterium Chromobacterium violaceum
 Former Flavobacterium  Found in soil and water
 Yellow colonies on BAP  Has the ability to produce
VIOLECIN, a purple pigment.