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Gram Positive Bacilli

Spore forming Gram (+) bacilli Catalase


(+)
Bacillus
Clostridium
Non spore forming Gram (+) bacilli
Catalase (+)
Corynebacterium
Listeria
Non spore forming Gram (+) bacilli
Catalase (-)
Lactobacillus
Erysipelothrix
Bacillus
- aerobic, gram positive, spore
forming rods
- habitat: thermal springs, fresh
and salt water, soil, on plants
- mostly contaminants in clinical
specimens except Bacillus
anthracis
Spores

- Resistant to radiation, heat,


disinfections, and desiccation
- Produced when the bacteria
gets stressed (Drying,
Temperature extremes)
- Aid in organisms survival
- Gram stain: appear as clear
areas within the bacterial cell
- specimen processing will involve
heat or alcohol shock before plating
on solid media
- TSI agar, urea agar, nutrient agar
containing 5mg manganese sulfate
per liter
- malachite green
- 5% sheep blood agar, chocolate agar
- PEA
Bacillus anthracis
Agent of anthrax, a disease in livestock
Humans acquire infection by
contamination of wound or ingestion or
inhalation of spores
Bacillus cereus
Causes food poisoning
An opportunist
Bacillus subtilis
Common laboratory contaminant
Used in sterility testing
Bacillus anthracis
- reservoir: Hoof stock (sheep, cattle,
goats); Fur, hides (textile workers);
Soil contaminated with spores
- MOT: contact with infected animal or
products (Inhalation, Inoculation,
Ingestion meat)
- non-motile
- Virulence: capsule, spore ( central),
exotoxin
has two circular, extrachromosomal, double-
stranded DNA plasmids, pXO1 and pXO2
which are required for full virulence

pXO1: contains the genes that encode for


the anthrax toxin components: protective
antigen (PA), lethal factor (LF), and edema
factor (EF)

pXO2: encodes a five-gene operon which


synthesizes a poly--D-glutamic acid
(polyglutamate) capsule which protect itself
from phagocytosis
Cutaneous anthrax
Malignant pustule (also called black
eschar)
Organisms gain access through cuts;
localized infection
Majority of cases in the world are
cutaneous
Pulmonary/Inhalation anthrax
Acquired through inhalation of spores
May result in respiratory distress and
death

Gastrointestinal
Acquired by ingestion of contaminated
raw meat
Usually fatal
Anthrax does not spread directly from
one infected animal or person to
another; it is spread by spores which can
be transported by clothing or shoes. The
body of an animal that had active
anthrax at the time of death can also be
a source of anthrax spores. Owing to the
hardiness of anthrax spores, and their
ease of production in vitro, they are
extraordinarily well suited to use as
biological weapons.
Medium-large, gray, flat, irregular with
swirling projections (Medusa head),
non hemolytic
Ascoli's test

- A precipitin test for anthrax using a


tissue extract and anthrax
antiserum.
Goal in identification is to RULE OUT B.
anthracis
If B. anthracis is suspect, MUST work
under safety hood
Bacillus cereus
- Most commonly encountered species of
Bacillus in opportunistic infections
- Usually involve intravenous drug abusers
or immunocompromised patients
- Virulence: enterotoxins, pyogenic toxins
- Motile, aerobic, NE
- Produces wide zone of lecithinase
on egg yolk agar
Food poisoning

Diarrheal form Emetic form


Associated with Associated with
meat, poultry, fried rice
and soups Abdominal cramps
Incubation period and vomiting
of 8 to16 hours Incubation period
Fever uncommon of 1 to 5 hours
Resolves within 24 Resolves in 9
hours hours
Local infections
Postsurgical/traumatic wounds
Burns
Eye infections

Rare conditions
Meningitis
Bacteremia
Endocarditis
Osteomyelitis
Bacillus subtilis
Found in the environment
Common laboratory and hospital
contaminant
Location: Soil, Decaying organic matter
Nonpathogenic: Research organism
(antibiotics, enzyme production,
insect toxin, fungicide, QC agent for
sterilization procedure)
Food spoilage
Industrial significance
A portion of the Bacillus thuringiensis
genome was incorporated into corn
(and cotton) crops. The resulting
GMOs are therefore resistant to some
insect pests.
Clostridium
Anaerobic, G+, motile rods
Found in soil, animal feces.
Spores is placed centrally,
subterminally or terminally; most
species are motile with flagella
Obligate anaerobes
Clostridium perfringens
Membrane disrupting
toxin
- Alpha toxin
(lecithinase)
Rupture RBCs,
Destroy Tissue
Exposure
Puncture wounds,
fractures
Surgical
Contaminated food
Diseases
Food poisoning
(Improper cooking)
Gas gangrene
Gas Gangrene

Treatment
Debride
AB
Hyperbaric O2
Nagler's Reaction: the suspect
organism is cultured on an egg yolk
media plate. One side of the plate
contains anti-alpha-toxin, while the
other side does not. An area of
turbidity will form around the side
that does not have the anti-alpha-
toxin, indicating uninhibited
lecithinase activity.
Clostridium tetani
Epidemiology:
Soil, Dust, GI tract,
Wounds (deep)
Burns, Umbilical
stump, Animal
bites
Tetanospasmin
(exotoxin)
Tetanospasm
Localized (Lockjaw)
Cephalic
Generalized
Death
respiratory failure
Dx
Spores in wound
Blood toxin levels
Toxin neutralization
test
Treatment
Antitoxin
AB
Debride
Curare poison
paralysis
ventilator
Prevention
Vaccination
Toxoid (DPT)
Antitoxin
Characteristics:
- box-car shaped
- anaerobic
- Gram-positive
- Gram stain: tennis rackets or
drumsticks
Tetanus neonatorum
Umbilical cord infected
soil, dirt rubbed on stump
Dirty instruments
Clostridium botulinum
Location
Soil, Water, Improper
canning, GI tracts of
animals
Epidemiology
Food borne
Toxin (canned
food)
Infant
Ingestion of
endospores
Source: honey
Wound
endospores
Pathogenicity
Botulinum toxin
Bind to neuron
prevent ACH
from binding
at NMJ
Flaccid
paralysis
Descending
Death in 3-
7 days
Dx
Culture/Isolation
Symptoms
Tx
Antitoxin
AB: penicillin
Ventilator
Prevention
Proper food handling
Boiling 10 minutes
Clostridium difficile
GI source
Epidemiology
Long term care facilities
Chronic AB use
Fecal-oral route
Pathogenicity
Toxins
Spores
Diseases
Colitis
pseudomembrane
Cecal perforation = fatal
- motile
- prevalent in soil
- Mx: long, irregular (often drumstick- or
spindle-shaped) cells with a bulge at their
terminal ends
- GS: Gram-positive, optimum growth on
blood agar at human body temperatures in
the absence of oxygen
- When stressed, the bacteria produce
spores that can tolerate extreme conditions
that the active bacteria cannot tolerate
C. sordellii: fatal infection in
exceptionally rare cases after
medical abortions.
Less than one case per year has been
reported since 2000.
Corynebacterium
Characteristics
Club shaped,
Metachromic granules
Epidemiology
Source: carriers
Cutaneous
Fomites
Respiratory infection
Aerosol
Pharynx, LN, tonsils
Pseudomembrane
Pathogenicity
Diptheria toxin
A: Inactivates proteins
B: attacks heart, NS
Neuraminidase
Protease (Dipthin)
Corynebacterium
diphtheriae
Infects nasopharynx or skin
Mostly grow aerobically, non-motile
Irregular swellings/clubbed-shaped;
granules within rod stained with aniline
dyes.
Diptheria
Pseudomembrane
Pharynx and
tonsils
Complications
(toxins)
Edema
Myocarditis
Neuritis
Cutaneous rash
Blood agar+potassium tellurite, tellurite
is reduced intracellularly
Dx
Pseudomembran
e
Metachromic
granules
Tx
Antitoxin
AB
Prevention
Vaccination (DPT)
GPR, catalase positive, NSF, NM,
straight or slightly curved
Metachromatic granules are usually
present representing stored
phosphate regions
The bacteria group together in a
characteristic way, which has been
described as the form of a "V",
"palisades", or "Chinese letters"
grow slowly, even on enriched media
need biotin to grow
grow in Loeffler's medium, blood agar, and
trypticase soy agar (TSA)
They form small grayish colonies with a
granular appearance, mostly translucent, but
with opaque centers, convex, with continuous
In TSA, they can form grey colonies with black
centers and dentated borders that look similar
to flowers (C. gravis), or continuous borders (C.
mitis), or a mix between the two forms (C.
intermedium).
Pathogenesis is based upon: (1) the ability of a
given strain of C diphtheriae to colonize in the
nasopharyngeal cavity and/or on the skin, and
(2) its ability to produce diphtheria toxin.
Pathology: Dt absorbed into mucous
membranes, causing destruction of epith and
superficial inflammatory response. Necrotic
epith embedded in exuding fibrin+RBC+WBC=
grayish pseudomembrane
Disease is principally result of the action of
toxin formed rather than invasion by the
organism
Vaccination confers protection against
disease by production of antibodies to the
diphtheria toxin. The vaccine is produced
from purified inactivated toxin from a
strain of C. diphtheriae.
Diphtheriae antitoxin (DTP) administered
to infants, and followed by boosters.
Treatment rapid suppression of toxin-
producing bacteria by antimicrobial drugs
at the earliest diagnosis of diphtheria.
Characteristics
Listeria
Soil, mammal gut
Non caspule, NSF
Motile [petrichous flagella]
Catalase (+)
Resist: Cold, pH, temp, salt, bile
Transmission Vertical
Direct: neonate, uterus
Dz
Food poisoning (dairy)
Tx: AB
Prevention
Cook foods
Pasteurize dairy products
Small/Short, G+,peritrichous flagella, rod.
Grow on Mueller-Hinton agar (better in
sheep blood-small zone of haemolysis);
facultative anaerobe, motile at room temp,
catalase +
Listeriosis results from ingestion of
contaminated food such as cheese and
vegie; primarily affects pregnant women,
newborns, those with weakened immune
system.
Able to multiply at low temp,hence
accumulate in contaminated food stored in
refrigerator.
Food Safety
Lactobacillus
Characteristics
Coccobacillus
FA microaerophilic
Lactic acid fermentation
Epidemiology
Food: meat, dairy, fruit
Environment: water
Food production
- Fermented vegetables
-Beverages: beer, wine, juices
- Milk products: yogurt, cheese
Problems
Food spoilage
also called Dderlein's bacillus
Gram-positive facultative anaerobic or
microaerophilic rod-shaped bacteria
major part of the lactic acid bacteria group, named
as such because most of its members convert
lactose and other sugars to lactic acid
In humans they are present in the vagina and the
gastrointestinal tract
They are usually benign, except in the mouth where
they have been associated with cavities and tooth
decay
The production of lactic acid makes its environment
acidic, which inhibits the growth of some harmful
bacteria.
Lactobacilli, especially L. casei and L.
brevis, are some of the most
common beer spoilage organisms.
Erysipelothrix
Characteristics
Saphrophyte
Nonmotile, NSF
microaerophilic
Epidemiology
Water, sewage
Animals
Pathogenicity: capsule
Transmission: Fecal
Disease
Macropapular rash
Endocarditis
Septicemia
Tx
AB
Animal vaccination
(pigs)
Erysipelothrix rhusiopathiae
Gram-positive, catalase-negative, rod-shaped,
non-spore-forming, non-acid-fast, non-motile
It may be isolated from soil, food scraps and
water contaminated by infected animals.
It can survive in soil for several weeks. In pig
faeces, the survival period of this bacterium
ranges from 1 to 5 months.
primarily considered an animal pathogen,
causing a disease known as erysipelas in
animals and erysipeloid in humans
Please read:
Proprionibacterium
Actinomyces
Streptomyces
Nocardia
Dont forget to wash your
hands!

YO U
H A N K
T
1. Enumerate 6 gram positive bacilli
2. Agent of anthrax
3. 3 forms of anthrax
4. Agent previously called Neisseria catarhalis
5. Agent with characteristic chinese letter
formation
6. Agent found in yogurt
7. Agent responsible for erysipeloid
8. Agent responsible for meningococcemia
9. Can be used as biological warfare weapon
10.4 medically important species of
Clostridium
Midterm Exam Coverage
10- Prelims
10- Branching G+ bacilli
25- G+ Cocci
25- G- Cocci
30- G+ Bacilli
100- Total

Date of Exam: January 31, 2015; 1-2pm;


7th floor
1. Enumerate 6 gram positive bacilli
2. Agent of anthrax
3. 3 forms of anthrax
4. Agent of Pseudomembrane colitis
5. Agent with characteristic chinese letter
formation
6. Agent found in yogurt
7. Agent responsible for erysipeloid (include
species name)
8. Agent for listeriosis (include species name)
9. Can be used as biological warfare weapon
10.4 medically important species of
Clostridium bacilli