Chapter 16: AEROBIC GRAM-POSITIVE BACILLI Clinical Infections - Carried in the

upper
Non-Spore Forming Non-Branching Catalase- respiratory tract
Positive Bacilli and spread by
droplet
Corynebacterium infection or
- Gram Stain: “club shape”, slightly curved, gram- hand-to-mouth
positive rods with unparallel sides and slightly contact
wider ends. - Most common

Respiratory Diphtheria
- Mostly found as normal biota on skin and mucous site of infection:
membranes of humans and animals tonsils or
- Divided into (1) nonlipophilic and (2) lipophilic pharynx
 Lipophilic corynebacteria are fastidious
and grow slowly and must be incubated - Cause tissue
at least 48 hours before growth is necrosis and
detected exudate
 Growth is enhance if lipids are included formation
in the medium - exudate forms a
There are more than 80 species in the genus, and tough gray-to-
at least 50 are thought to be clinically significant white
*liphophilic – fat loving bacteria pseudomembran
`Corynebacterium spp. can be opportunistic e. Appear on the
pathogens, especially in immunocompromised tonsils and
patients. spread
downward into
the larynx and
trachea
- Consist of non-

Cutaneous Diphtheria
healing ulcers
with a dirty gray
membrane.

Corynebacterium diphteriae
Virulence Factors - Diphtheria Toxin
- Major virulence factor
- Produced by strains
Only toxin- infected with a Drug of Choice - Penicillin
producing C. lysogenic beta-phage, - Erythromycin (for
diphtheriae which carries the tax penicillin-sensitive
causes gene for diphtheria toxin individual)
diphtheria - Block protein synthesis
`C. ulcerans and in eukaryotic cells Laboratory Diagnosis
C. - Composed of two
pseudotuberculo fragment: Microscopy - Highly pleomorphic gram-
sis can produce  Fragment A – positive bacillus
the toxin when responsible for - Appears in palisades or
they become cytotoxicity as individual cells lying at
infected  Fragment B – sharp angle to another in
the toxin is binds to receptors “V” or “L” formations
nontoxic until on the eukaryotic - Babès-Ernst granules –
exposed to cells ; mediates the metachromatic areas of
trypsin which entry of fragment A the cell
cleaves the toxin into the cytoplasm. o Stained with
into two methylene blue
fragments

Culture - Facultative anaerobe

Characteristics - Grows best under
aerobic conditions and
pleomorphi has on optimal growth
c – many temperature of 37 deg.C
shapes - Multiplication occurs
`Better within 15 deg.C to 40
growth is deg.C
 obtained on - Can have a very small appearance, and
a medium zone of -hemolysis on nonlipophilic
containing Loeffler medium.
blood or - Black or brownish Corynebacterium - Most common
serum such colonies with brown halo jeikeium cause of
as Loeffler surrounding the colony Corynebacterium-
serum or from the reduction of associated
Pai agars. tellurite on Cystine- prosthetic valve
`CTBA – tellurite blood agar endocarditis in
selective (CTBA). adults.
and - - Lipophilic, strict
differential aerobe,
medium; nonhemolytic, does
can be not produce urease,
differentiate and is nitrate-
d with staph production negative
and strep - Drug of choice:
because of VANCOMYCIN
brown halo Corynebacterium - Part of the normal
on CTBA pseudodiphtheriticum biota of the human
nasopharynx
Identification - Catalase-positive and - Frequently
non-motile associated with
- Lack of urease respiratory tract
production infections in
- Ferments glucose and immunocompromised
maltose individuals with other
- Produce acid but not underlying diseases.
gas - Can mimic
- Reduces nitrate to respiratory diphtheria
nitrite - Reduces nitrate and
Test for - Elek Test produces urease
Toxigenicity o Immunodiffusion test Corynebacterium - Causes a
o In vitro diphtheria pseudodiphtheriticum granulomatous
C. diphtheriae toxin detection lymphadenitis in
is - Rapid enzyme-linked humans
distinguished immunosorbent assays - Produces a
from the other - Immunochromatograph dermonecrotic toxin
two species ic assays and can produce
by its lack of - Polymerase Chain diphtheria toxin
urease Reaction - Produces brown halo
production o Detection of tax on CTBA, produces
`Diagnosis of gene urease and appears
diphtheria is small yellowish white
established by colonies on SBA
also Corynebacterium - Part of human skin
demonstrating striatum and nasopharynx
that the microbiota
isolate - Commonly
produces associated with
diphtheria device-related
toxin infection
- Nonlipophilic,
pleomorphic and
Other Corynebacteria produces small,
shiny, convex
Corynebacterium - Often associated colonies
amycolatum with prosthetic joint Corynebacterium - Isolated from
infection, blood ulcerans humans with
stream infections and diphtheria-like
endocarditis in illness, skin ulcers
immunocompromised and exudative
patients or patients in pharyngitis
a health care setting. - Human infections
- Flat, dry, have a acquired through
matte or waxy contact with animals
 or by ingestion of spontaneous
unpasteurized dairy abortion and
products stillborn neonates
- Produces brown halo - Flu-like illness with
on CTBA, narrow fever, headache,
zone of -hemolysis and myalgia
on SBA, does not - May progress and
reduce nitrate and is result in premature
urease-positive labor or septic
Corynebacterium - Commonly abortion within 3 to 7
urealyticum associated with days
UTIs; urine isolates Disease in the Newborn - Two forms of
shows pinpoint, NEONATAL
nonhemolytic white LISTERIOSIS:
colonies - Early onset –
- Lipophilic and strict intrauterine infection
aerobe, nitrate- that can cause
negative, catalase- illness shortly after
positive, rapidly birth. The result is
urease-positive most often sepsis. It
within minutes after may be associated
inoculation on a with aspiration of
Christensen urea infected amniotic
slant fluid; most often
- Drug of choice: sepsis
VANCOMYCIN - Late onset – occurs
several days to
C. amycolatum and C. jeikeium part of the normal weeks after birth.
skin microbiota The disease is most
`C. pseudodiphtheriticum normal biota of human likely to manifest as
nasopharynx meningitis.
`C. pseudotuberculosis is primarily a veterinary
pathogen; human contact with sheep Disease in the - INVASIVE
`dermonecrotic toxin – causes death of various cell Immunosuppressed LISTERIOSIS most
types Host common
manifestations are
Listeria monocytogenes CNS infection and
- Recovered from soil; water; vegetation; and animal endocarditis.
products, such as raw milk, cheese, poultry, and - Diagnosis is made
processed meats. by culturing L.
- has also been isolated from crustaceans, flies, and monocytogenes
ticks. from the blood or
- Listeriosis is recognized as an uncommon but CSF.
serious infection primarily of neonates, pregnant - Drug of choice:
women, elderly adults, and immunocompromised AMPICILLIN
hosts.
Laboratory Diagnosis
Virulence Factor Microscopy - Gram-positive
 Hemolysin (Listeriolysin O) coccobacillus
o Damages the phagosome membrane, effectively Culture Characteristics - SBA, chocolate
preventing killing of the organism by the agars, nutrient agars
macrophage and in broths such
 Catalase as brain-heart
 Superoxide dismutase infusion and
 Phospholipase C thioglycolate
 Surface protein (p60) - Colonies are small,
o Induces phagocytosis through increased round, smooth, and
adhesion and penetration into mammalian cells translucent.
- Surrounded by
Clinical Infections narrow zone of -
Disease in Pregnant - LISTERIOSIS is hemolysis which is
Women most commonly visualized when
seen during the 3rd colony is removed
trimester - Optimal growth
- L. monocytogenes is temperature is 30C
responsible for to 35C, but growth
 occurs between Clinical Infections
0.5C to 45C - Localized
- Cold enrichment – skin disease
technique used to - Lesions are
isolate the organism usually seen
from clinical on hand or
specimens fingers
Identification - Catalase-positive,

ERYSIPELOID
- INCUBATION
motile at room PERIOD: 2 to
temperature, - 7 days
hemolytic, and - Self-limiting
hydrolyzes esculin infection that
- Exhibits tumbling normally
motility, heals within 3
microscopically to 4 weeks
- Exhibits umbrella but may
pattern of growth in continue for
motility medium at months
room temperature, -
22C to 25C - Often
- Hippurate associated
hydrolysis, bile with

SEPTICEMIA
esculin hydrolysis, endocarditis
and CAMP test that has been
positive (“block” seen in
type hemolysis on patients who
CAMP test) have had
- Confirmatory test: valve
acid production from replacements
glucose, positive -
Vogues-Proskauer - Rare and

CUTANEOUS INFECTION
GENERALIZED, DIFFUSE
and methyl red manifests as
reactions an
exacerbation
of the
Non-Spore Forming Non-Branching Catalase- erysipeloid
Negative Bacilli lesion

Erysipelothrix rhusiopathiae

General - Gram-positive, Antimicrobial therapy is effective for

Characteristics catalase-negative,
non-spore forming,
pleomorphic rod that
has a tendency to
form long filaments
Laboratory Diagnosis
- Usual route of
infection: cuts or
MICROSCOPY
scratches on the skin
- Organism is resistant
to salting, pickling
and smoking
- Survives well in
water, soil, and plant
material
CULTURE
CHARACTERISTICS
treating E. rhusiopathiae infections
Resistant to VANCOMYCIN
DRUG OF CHOICE: PENICILLIN
- Thin, rod-shaped,
gram-positive
organism that can
form long filaments
- arranged singly, in
short chains, or in a
V shape
-
- Isolation of the
organism include
tissue biopsy or
aspirates from skin
lesions
- Should be
inoculated to a
nutrient broth with
1% glucose and
 incubated in 5% VAGINOSIS in
CO2 at 35C humans
- Grows on standard o BV is
culture media such characterized by
as SBA and a malodorous
chocolate agar discharge and
o On SBA, colony vaginal pH
appearance is greater than 4.5
usually o It results from
nonhemolytic and reduction in the
pinpoint after 24 Lactobacillus
hours of incubation population in the
o After 48 hours of vagina, followed
incubation, two by an increase in
distinct colony vaginal pH; this
types are seen: (1) results in
A smaller, smooth overgrowth by G.
form is vaginalis and
transparent, other BV-
glistening and associated
convex with entire organisms
edges; (2) larger, - G. vaginalis also
rough colonies are play a role in UTIs in
flatter with a matte men and women
surface, curled DRUG OF CHOICE - Metronidazole
structure and
irregular edges Laboratory Diagnosis

IDENTIFICATION - Catalase-negative, MICROSCOPY - Often appears

nonmotile, pleomorphic, gram-
pleomorphic, aerobic variable
or facultatively coccobacillus or
anerobic gram- short rod
positive rod, - CLUE CELLS –
hydrogen sulfide large squamous
positive, urease and epithelial cells with
Vogues-Proskauer gram-positive and
negative, and does gram-variable bacilli
not hydrolyze and coccobacilli
esculin. CULTURE - Vaginal discharge is
- “test tube brush-like” CHARACTERISTICS the most common
in a gelatin stab specimen used for
culture at 22C isolation
- Grows best in 5% to
7% CO2 at a
temperature of 35C
to 37C
- On SBA, appears
Gardnerella vaginalis pinpoint,
nonhemolytic
GENERAL - Normal biota in the colonies
CHARACTERISTICS human urogenital - MEDIUM OF
tract CHOICE: Human
- Short, pleomorphic Blood Bilayer Tween
gram-positive rod or (HBT) Agar
coccobacillus - Colonies are -
- Has a gram-positive hemolytic when
type of cell wall but cultured on human
the peptidoglycan and rabbit blood, but
layer is thinner than not on sheep blood
that found on other IDENTIFICATION - Catalase-negative,
gram-positive oxidase-negative,
bacteria and Hippurate
CLINICAL INFECTIONS - Associated with hydrolysis-positive
BACTERIAL
 Non-Spore Forming, Branching Aerobic
Actinomycetes MICROSCOPY - Gram-positive,
beaded branching
Nocardia filaments
- Partially acid-fast on
GENERAL - Aerobic, branched, stain with
CHARACTERISTI beaded gram-positive carbolfuchsin and
CS bacilli decolorizing with a
- Modified acid-fast positive weak acid (0.5% to
- Infections generally occur 1% sulfuric acid)
in immunocompromised - SPECIMEN OF
patients CHOICE: Tissue
VIRULENCE - Produce superoxide and Pus from the
FACTORS dismutase and catalase draining sinuses
that may provide - The granules can be
resistance to oxidative visualized by
killing phagocytes separating them
- They also produce an from the pus with an
iron-chelating compound inoculating needle
called nocobactin. and then washing in
Clinical Infections - Mostly caused by sterile saline
- Found in soil N. -
and on plant cyriacigeorgica CULTURE - Oxidative-type
PULMONARY INFECTIONS

material and N. farcinica CHARACTERISTICS metabolism; they

- Infection - Confluent use a wide variety of
occurs by two bronchopneumo carbohydrates
routes: (1) nia that is usually - Grows well on most
pulmonary and chronic but may common non
(2) cutaneous be acute or selective laboratory
- Mortality rate is relapsing media incubated at
high, and - The initial lesion in 22C to 37C
patients who the lung is a focus although 3 to 6 days
survive often of pneumonitis or more may pass
have significant that advances to before growth is
tissue damage necrosis seen
- The sputum is - Selective media:
thick and purulent Modified Thayer-
- N. brasiliensis is Martin
the most frequent - Chalky, matte,
form of velvety, or powdery
nocardiosis – appearance and
usually seen in the may be white,
hands and feet as yellow, pink, orange,
a result of outdoor peach, tan or gray
activity pigmented.
CUTANEOUS INFECTIONS

- Occurs after - Dry, crumbly

inoculation of the appearance similar
organism into the to breadcrumbs
skin or TREATMENT - Resistant to
subcutaneous penicillin but
tissues susceptible to
- Infection begins as sulfonamides
a localized 3% HCl in the stain for mycobacteria
subcutaneous `often seen in sputum and exudates or aspirates
abscess that is from skin or abscesses
invasive which are
termed as Other Actinomycetes
actinomycotic Actinomadura - Etiologic agents of
mycetomas mycetomas
- Pus may be Streptomyces - Primarily
pigmented and saprophytes found
contain “sulfur as soil inhabitants
granules” - Streptomyces
- somaliensis is an
established human
Laboratory Diagnosis pathogen associated
 with actinomycotic -
An adenylate

Edema Factor
mycetoma cyclase that
Gordonia - Aerobic, catalase- increases the

(EF)
positive, gram- concentration of
positive to gram- cyclic adenosine
variable, partially monophosphate
acid-fast, and (cAMP) in host
nonmotile cells
- Gordonia bronchialis - A protease that

Factor (LF)
associated in kills host cells by

Lethal
postsurgical sternal disrupting the
wounds, coronary transduction of
artery infection, and extracellular
central venous regulatory signals
catheters CLINICAL - Woolsorter’s disease and
Tsukamurella - Gram-positive, INFECTIONS Ragpicker’s disease
aerobic, catalase- - Humans are o Were used to describe
positive, and infected infection with the
partially acid-fast primarily as a spores of B. anthracis
- Most infections are result of as a result of handling
caused by accidental or contaminated animal
Tsukamurella occupational fibers, hides, and
paurametabola exposure to other animal products
Rhodococcus - Rhodococcus equi – animals or - Three main forms of
most common animal anthrax are recognized in
human isolate products humans:
- Salmon-pink - It is a 1. Cutaneous
pigment that bioterrorism 2. Inhalation or
resembles Klebsiella agent Pulmonary
Tropheryma whipplei - Causative agent of 3. Gastrointestinal
Whipple’s Disease - A fourth form has emerged
- First identified by recently, called injectional
PCR from a anthrax
duodenal biopsy
specimen anthrax toxin consists of 3 proteins; individually
Spore Forming Non-Branching Catalase-Positive nontoxic but act synergistically together
Bacilli to produce damaging effects
`the effect of EF and LF is seen when combined
Bacillus with PA
- Gram-positive or gram-variable OTHER CLINICAL INFECTIONS
- Aerobic or facultative anaerobic bacilli that form CUTANEOUS - Occur when wounds
endospores ANTHRAX are contaminated
- Some species are thermophiles that grow best at with anthrax spores
55C or higher acquire through skin
- Most species grow well on SBA cuts, abrasions or
- Non pigmented colonies insect bites
- Catalase-positive and form endospores under - 2 – 3 days after
aerobic and anaerobic conditions exposure, a small
Bacillus anthracis pimple or papule
appears at the site
VIRULENCE - Protects the of inoculation
CAPSULE

FACTORS organism from - Ring of vesicles

- Depends on a phagocytosis develops and
glutamic acid coalesce to form an
capsule and a erythematous ring
three- - Binding molecule - Small dark area
Antigen (PA)

component for EF and LF, appears in the

Protective

protein permitting their center of the ring

exotoxin attachment to and eventually
specific receptors ulcerates and dies,
on the host cell’s forming a depressed
surface black necrotic
central are known as
an eschar or black
eschar (1-3 cm in
diameter)
 INHALATION ANTHRAX - Also called
WOOLSORTER’S
DISEASE, is
acquired when
spores are inhaled
into the pulmonary
parenchyma
- 2 – 5 days after
exposure to the
spores, a
nonspecific illness
consisting of mild
fever, fatigue and
malaise
- Sudden severe
phase, respiratory
distress
GASTROINTESTINAL - Occurs when the
ANTHRAX spores are
inoculated into a
lesion on the
intestinal mucosa
after ingestion
- Symptoms include
abdominal pain,
nausea, anorexia,
bloody diarrhea, and
vomiting
INJECTIONAL - Characterized by
ANTHRAX soft tissue infection
associated with
“skin popping” or
other forms of
injection drug use
and results from the
direct injection of the
spores into tissue
- Can be associated
with necrotizing
fasciitis, organ
failure, shock, coma, - B. cereus is a relatively common cause of food
and meningitis.
`cutaneous: lesions are painless and does not
produce pus. Begins to heal after 1-2weeks - Takes two forms: (1) diarrheal and (2) emetic
Laboratory Diagnosis
MICROSCOPY - Large, square-
ended, gram-
positive or gram- - Can be grown aerobically at 37C on SBA
variable rod found - A -hemolytic frosted glass-appearing colony
singly or in chains
- “bamboo-rod”
appearance, ends of
the single cells fit
snugly together with
also an unstained
central spore
CULTURE - On SBA, colonies of
CHARACTERISTICS B. anthracis are
nonhemolytic, large,
gray and flat with an
irregular margin
because of
outgrowths of long
filamentous
projections
- “Medusa head”
appearance has
been used to
describe the colony
morphology of B.
anthracis
IDENTIFICATION - Catalase-positive,
grows aerobically
and anaerobically;
nonmotile
- Ferments glucose,
fails to ferment
mannitol, arabinose
or xylose; Produces
lechithinase
- Capsule production
can be detected by
India ink stain on
blood or CSF
-
TREATMENT - Mostly susceptible to
penicillin but
resistance can occur
in the absence of -
lactamase
production
- Ciprofloxacin and
doxycycline used for
initial intravenous
therapy
- Addition of
clindamycin is
recommended to
inhibit exotoxin
production
Bacillus cereus
poisoning and opportunistic infections in
susceptible hosts
o Diarrheal syndrome is usually associated with
ingestion of meat or poultry, vegetables and
pastas by an incubation period of 8 to 16
hours
containing spore-forming, gram-positive bacilli that
are motile, able to ferment salicin, and lecithinase-
positive