1. All of these organisms produce the pigment pyoverdin
2. Ubiquitous in distribution
3. Opportunistic pathogens

1. Commercial gas-generating systems
2. Evacuating-replacement systems
3. Polyethylene bag
4. Candle jar

• Long, thin gram-negative rod
• grows on BAP and is often beta-hemolytic
• grows on MAC and is a non-fermenter
• spready, flat colonies with serrated edges
• bluish-green, red, or brown pigmentation
• often shows as metallic sheen
• grape-like or corn taco-like odors

• Refrigerated if unable to process within 30 minutes
•  Frozen at -70 C if unable to process within 24 hours

1. Respiratory Tract Infections
2. "Swimmer's ear"
3. Wound infections in Burn patients
4. UTI's
5. Eye infections
6. Respiratory infections in CF patient

(Opportunistic pathogen)

Bacteria that produce enzyme tryptophanase are able to degrade the amino acid tryptophan into pyruvic acid, ammonia, and indole.

When 1% paradimethylaminocinnamaldehylde is added, pink color occurs

Bordet-Gengou agar

Charcoal-horse blood agar (aka Regan-Lowe agar)

Modified Jones-Kendrick charcoal

Causes chronic gastritis (duodenal and peptic ulcers)

Able to survive the low pH of the stomach by creating its own alkaline environment

Produces urease that hydrolyzes urea to form ammonia to neutralize the stomach acid

Characteristics:

• gram-negative rods
• ubiquitous in distribution
• opportunistic pathogens
• obligate aerobes
• slow growers 
• Oxidase positive
• non-lactose fermenters
• typically non-motile

Cholera toxin or choleragen

 Causes mucosal cells to hypersecrete electrolytes and water into the lumen of GI tract which leads to rice-water stools. Can lead to severe dehydration, hypovolemic shock, metabolic acidosis, and death in a matter of hours.

Collection from stool specimens are preferred.

Transported via Cary-Blair medium

Rectal swabs are acceptable

Culture from endoscopy specimen

Histologic analysis of biopsy

Campylobacter-like organism (CLO) Test [rapid urease test]

Direct Fluorescent Antibody (DFA)

Polymerase Chain Reaction (PCR)

Serological diagnosis

Dry, wrinkled, buff to brown in color colonies.

Non-lactose fermenter

Fastidious, gram-negative organism

Shows a weak catalase positive

Also shows a weak oxidase positive reaction.

Does not ferment or oxidize carbohydrates

Hippurate positive.

Susceptible to azithromycin, fluroquinolone, or doxycycline.

Ferments flucose

Oxidase negative

Reduces nitrate to nitrite

Fluid and electrolyte replacement

Antimicrobial therapy: tetracycline

Gram-negative coccobacilli
Requires both Factors X and V 

Shows no Hemolysis on Rabbit Blood
Catalase positive
Oxidase positive
ALA Test negative
Causes purulent conjunctivitis (pink eye) and Brazilian purpuric fever 

Capsular characterisitics

IMViC = ++- +/-

Lactose nonfermenter

Ornithine negative

Urease positive

Swarming

H₂S positive

PDA positive

IMViC = +-++

DNase negative

Ornithine negative

Lactose fermenter

H₂S negative

Citrate negative

IMViC = -+ +/- +/-

PDA positive

Lactose non-fermenter

H₂S positive

Urease positive

Ornithine positive

Swarming

IMViC = --++

DNase negative

Lactose fermenter

Citrate positive

Ornithine positive

Lysine negative

Motile

IMViC = --++

H₂S negative

citrate positive

lactose fermenter

DNase negative

ornithine negative

nonmotile

Lactose Non-Fermenter

IMViC = ++--

H₂S negative

PDA negative

Citrate negative

Motile at 22 C

Lactose Non-fermenters

IMViC = ++--

PDA negative

H₂S negative

Non-motile at 22 C

Citrate negative 

Lactose fermenter

IMViC = -+-+

Urease positive

H₂S positive

Ornithine negative

DNase negative

Lactose fermenter

Ornithinie positive

Lysine positive

DNase negative

IMViC = --++

motile

Lactose non-fermenter

H₂S negative

IMViC = ++-+

PDA positive

Lactose non-fermenter

H₂S negative

PDA positive

IMViC = ++--

Lactose non-fermenter

IMViC = ++--

H₂S positive

Urease negative

Lactose nonfermenter

H₂S positive

Urease negative

IMViC = -+-+

Lysine positive

Microaerophilic environment (5 - 10% oxygen)
Capnophilic environment (8 - 10% oxygen)

Needs at least 48 - 72 hours for growth 

Modified Skirrow's media

Campy-BAP

Campy-CVA

Campy Thio

Motile

Lactose Fermenter

Produce gas from glucose

Ornithine positive

IMViC = --++

Non-lactose fermenter

H₂S negative

Citrate negative

PDA negative

Non-motile

IMViC = -+--

Open tube yellow; closed tube yellow = Fermenter

Open tube yellow; closed tube green/blue = Oxidizer

Open tube green/blue; closed tube green/blue = Non-utilizer

Organisms are lactose negative

PDA positive 

H₂S positive

IMViC = --+/- +/-

Ornithine positive

Urease positive

Organisms of this genus are indole negative

Late or slow lactose fermenter

DNAse positive

ONPG positive

Ornithine positive

Lysine positive

Citrate positive

VP positive

-+++

Oxidase positive

TSI = Alk/Alk

Bluish green, red, or brown pigment on Mueller-Hinton or TSA

Grows at 42 C

Pink = positive

No color change = negative

Reagent: 1%-paradimethylaminocinnamaldehyde 

Positive = bright pink or red color

Negative = no color change

Positive = green color

Negative = no color change

Ferric chloride = reagent for PDA

Positive = purple

Negative = yellow

Positive = purple slant and butt (lysine decarboxylation)

Positive = red/plum slant and purple butt (lysine deamination)

Negative = purple slant and yellow butt (negative for lysine decarboxylation)

Negative = red slant/yellow butt (negative for lysine deamination) 

Reservoirs of this organism include plants, soil, and water.
It is intrinsically resistant to multiple antimicrobial agents.

It can survive medical devices and solutions (including disinfectant solutions)

Infections result from direct contact with contaminated food, medical solutions, and respiratory equipment

Stool and rectal swabs

Cary-Blair transport medium if unable to be cultured within 4 hours of collection

Blood specimen (as opposed to another species with similar methods)

Sucrose fermenters = yellow

Non-sucrose fermenters = green

These organisms are generally curved, gram-negative rods. They can appear small, straight rods which can be highly pleomorphic.

This fastidious organism shows to be gram-negative rods
Catalase positive

Oxidase positive

Shows hemolysis on Rabbit blood

Requires both factors X and V

ALA Test negative 

This gram negative bacillus organism:

is H₂S positive

Urea positive in 1 - 2 hours

Some strains require CO₂

Found in cattle

This gram negative coccobacilli organism is nonmotile.

Oxidase positive

Catalase positive

Acidic in butt of TSI

Urea negative

Nitrate positive

Infole positive

Ornithine positive

Susceptible to penicillin

This organism is Gram-negative coccobacilli 

Fastidious

Catalase negative

Oxidase negative

Requires Factor X (hemin) 

ALA Test negative 

This organism is a lactose fermenter

It is oxidase negative

ONPG positive

Lysine positive

Produces gas from glucose 

IMViC = ++--

This organism is fastidious and gram-negative pleomorphic rods

ALA Test positive

Shows no hemolysis on Rabbit blood

Requires only Factor V

May be catalase positive

May be oxidase positive

This organism is gram-negative coccobacilli.
Requires both Factors X and V
Shows no hemolysis on Rabbit Blood
Catalase positive

Oxidase positive
ALA Test negative 

This organism is lactose negative

PDA positive

H₂S positive

urease positive

ornithin negative

IMViC = ++- +/-

This organism is nonmotile

Usually oxidase postive

Catalase positive

Some species oxidize glucose

Susceptible to tetracycline w/ or w/o Streptomycin

Gram-negative coccobacilli seen singly, pairs, or short chains

This organism is oxidase positive

Glucose oxidizer

Grows at 42 C

Reduces nitrate to nitrogen gas (4+ reaction)

All decarboxylase tests are negative

This organism is oxidase positive

Sucrose positive on TCBS

Lactose negative on MAC

Lysine positive

Ornithine positive

VP negative

String Test positive

This organism is small, gram-negative rods

Colonies are convex and may appear sulfur yellow or may be colorless.

Motile

Oxidase negative (or weak positive)

Glucose oxidizer

Lysine positive

Ornithine positive

Arginine negative

This organism is transmitted via the following ways:
1. Fecal-oral route

2. Contact with contaminated water

3. Ingestion of contaminated shellfish or other seafood 

This organism is very mucoid (due to encapsulation)
Lactose positive

Non-motile

Urea positive

IMViC = +-++

DNAse negative

Ornithine negative 

This organism is:

Oxidase positive

Hippurate positive

Catalase positive

Cephalothin resistant

Naladixic Acid susceptible

This organism is:
Oxidase positive

Hippurate negative

Catalase positive

Cephalothin resistant

Naladixic Acid susceptible 

This organism shows:

Curved gram-negative rods

Comma shapes, "S" shapes and gull wing forms

Short or occasionally long chains

"Darting motility"

This organism was collected from bone marrow, blood, tissues, and joint fluid. 

Can be cultured on BAP and CHOC

Colonies are slightly yellow, convex, smooth, translucent, pearl-like, and incubated for 48 hours

Gram-negative coccobacilli in pairs or short chains

Stained with carbol-fuchsin

Grows in 8 - 10% CO₂

Causes undulant fever or Bang's disease

This Brucella species is found in goats.

Does not need CO₂

Is H₂S negative

Grows in the presence of thionin, thionin blue, and fuchsin

Brucella canis

This Brucella species is found in dogs.

Needs no CO₂

Urea positive in less than 30 minutes

Grows in presence of thionin, but not in fuchsin

This Brucella species is often found in swine

Does not require CO₂

Urea positive in less than 30 minutes

Most are H₂S negative

Most grow in presence of thionin, but most don't grow in presence of fuchsin

Will grow on BAP, CHOC, and MAC (non-fermenter)

Alkaline peptone water broth (enrichment)

Thiosulfate Citrate Bile Salts Sucrose (selective/differential)

lactose fermenter

DNase negative

IMViC = ++--

lactose fermenter

DNase positive

IMViC = -+++

pyocyanin (blue)

pyoverdin (yellow-green or green-brown)

pyorubin (red)

pyomelanin (brown)

Pyocyanin + Pyoverdin = characteristic "green" color 

21% oxygen

0.03% carbon dioxide

15% oxygen

5 - 10% carbon dioxide

These organisms: 

Do not grow in the presence of oxygen

Part of the human normal flora

Usually encountered as contaminants

Pathogenic infections are the result of invasion of a sterile site

True or False

Most anaerobic infections involve a mixture of anaerobic and facultative anaerobic organisms.

1) Trauma of mucous membranes or skin

2) Stasis

3) Tissue necrosis

4) Decrease in healing abilities of tissue

• Infection is in close proximity to a mucosal surface
• Presence of a foul odor
• Production of large quantity of gas
• Production of black exudates that fluoresces brick-red when exposed to long-wave U/V light
• Presence of sulfur granules
• Distinct microscopic characteristics in gram-stain preparations (morphology)

Body fluids other than random-collected urine

Suprapubic urine

Aspirates

Tissue

Bone marrow

Throat or nasopharyngeal swabs

Sputum or bronchial washings

Stool or rectal swabs (except C. difficile)

Vaginal, cervical or urethral

Voided urine

 specimens that have a 1) foul odor, appear to have much gas or 2) have a watery black exudate

(this way of examining allows you to suspect anaerobes)

Anaerobic BAP (AnBAP)

Bacteroides bile esculin (BBE)

Cycloserine-cefoxitin fructose agar (CCFA)

Egg yolk agar

Kanamycin-vancomycin laked blood (KVLB)

Phenylethyl alcohol blood agar (PEA)

Anaerobic broth

1. Sodium borohydride tablets

2. Sodium bicarbonate and citric acid

1) envelope gas generator

2) indicator

3) palladium catalyst

Sodium borohydride + water => Hydrogen
Hydrogen + Oxygen (via palladium catalyst) => Water 

Sodium bicarbonate + citric acid + water => CO₂

1) Anaerobic jars

2) Anaerobic bags

3) Evacuation-replacement procedure

4) Anaerobic chambers

5) Gloveless anaerobic chamber

1) Air from anaerobic jar is removed with vacuum

2) Replaced with nitrogen gas

3) Replace nitrogen with anaerobic gas mixture

1. Aerotolerance

2. Colony and gram-stain characteristics

3. Fluorescence

4. Commercial identification system

5. Disk tests

Causative agent of:

• Food-borne botulism
• Infant botulism
• Wound botulism 

Most common cause of antibiotic -associated diarrhea (AAD) and pseudomembraneous colitis.

This organism's colonies are yellow, ground-glass on Cycloserine-Cefoxitin Fructose Agar (CCFA). Odor resembles that of a horse stable. Fluoresce under UV light

Grows at 42 C
Needs at least 48 - 72 hours

Darting motility

Causative agent of gastroenteritis 

This organism requires body temperature (37 C)

Humidified atmosphere without elevated carbon dioxide

Demonstrates pale staining due to poor uptake of safranin

silver-gray and small colonies

Will grow on BAP

Colonies are convex, grey-white and non-hemolytic

"Hockey puck" colonies

Pinkish hue

No growth on MTM

Nitrate positive

Most are beta-lactamase positive

Asaccharolytic

MCAT Disk Screen - Butyrate esterase Positive (blue)

This organism inhabits soil.

It is transmitted via spore exposure

Gram Positive Bacillus

Accounts for 95% of anthrax cases

20% mortality if untreated

Also called Woolsorter's disease

Virtually 100% fatal if untreated

Gastrointestinal anthrax

Usually caused by ingestion of contaminated meat

Rapid onset of GI symptoms

Form a lesion on the intestinal mucosa

Caused by Bacillus anthracis.

- anti-phagocytic capsule

-exotoxin production: mediate cell and tissue destruction

- edema toxin

- lethal toxin

Non-hemolytic on blood

Large, flat, irregular colonies with swirling projections 

"medussa head" appearance

This organism is motile: tumbling motility on wet prep and umbrella shaped motility on semi-solid motility medium.

This gram-positive bacilli organism is:

catalase positive

hippurate positive

esculin positive

MR positive

CAMP positive

Grows in 4 C (cold enrichment)

CHEMC + 

They are usually gray to white, opaque, entire, convex colonies. 

Are pleomorphic, gram-positive rods. They occur in palisades, club-shapes, and Chinese letters.

This GPB is:

catalase positive

nitrate reduction positive

Elek test positive

This GPB is often carried by reptiles and other animals.

It is transmitted via puncture or wound of skin with the animal exposure.

Often associated with people who handle animals or animal products

True or False

Erysipeloid is a self-limiting disease

GPB is non-motile

May have long, non-branching filaments. 

May have short, thin rods in chains

Decolorizes easily

This GPB is:

catalase negative

oxidase negative

produces H₂S

Glucose positive

Lactose positive

beta-hemolysis on human blood Tween agar

hippurate positive

GPB

Chocolate - pinpoint tan colonies

small rods would attach to "clue cells"

This GPC grows well on BAP

Facultative anaerobes 

Incubation in 5 - 10% CO₂

Todd-Hewitt broth enhances recovery

GPC

Have mucopeptide and teichoic acid in cell wall

Cell wall also have C carbohydrate groups - antigenic

True or False

Only strains of C. diphtheriae infected with a bacteriophage are capable of producing the exotoxin

Contributes to tissue invasion and destruction

Responsible for hemolysis on BAP

Lyses leukocytes, platelets, and RBC's

oxygen labile

Highly antigenic

This toxin is non-antigenic

Lyses leukocytes

Is Oxygen-stabile

Causes skin infections: impetigo and erysipelas

Scarlet fever

Necrotizing fasciitis and mycostis "flesh-eating"

Rheumatic fever

Acute glomerulonephritis

Acute pharyngitis (strep throat)

Small

Transparent

Large zone of beta-hemolysis in relation to the colony size

Antistreptolysin O (ASO)

Anti-DNase B

Anti-streptokinase 

This organism causes:

Neonatal sepsis

Neonatal meningitis

Postpartum infections

Bacteremia

Skin and soft tissue infections

Capsule - prevents phagocytosis

CAMP factor

Lipotechoic acid

Protease

Deoxyribonuclease

Medium size

Gray-white

Creamy

Flat

Small zone of hemolysis in relation to the colony size

Polysaccaride capsule that inhibits phagocytosis

Pneumolysin

This GPC that shows partial hemolysis on BAP causes:

Pneumonia

Meningitis

Otitis media

Bacteremia

Microscopic examination:

Gram positive diplococci

Lancet shaped

Macroscopic examination:

Initially domed, then “dimpled”

Translucent

Alpha hemolytic

Encapsulated colonies

Larger

Mucoid or watery 

This organism causes:

Subacute bacteria endocarditis

Urinary tract infections

Septicemia with S. bovis highly associated with colonic 

carcinoma

Mixed wound infections

Similar to Enterococcus however these exhibit only alpha and gamma hemolysis 

Bile esculin positive

LAP positive

Bacitracin resistant

Hippurate negative

PYR negative

CAMP factor negative

No growth in 6.5% NaCl broth

Growth in 6.5% NaCl

Bile esculin – positive

PYR – positive