Bacteria Associated With Skin Infections 2 Flashcards
Aerobic bacteria associated with skin infections
Erysipelothrix
Bacillus
Anaerobic bacteria associated with skin infections
Clostridium
Most important species of Erysipelothrix
Erysipelothrix rhusopathiae
General principles of Erysipelothrix rhusiopathiae
Straight/slightly curved Gram (+) rods in singles, short chains V/L configuration Pleomorphic morphology Ubiquitous/abundant in nature Catalase negative
Determinants of pathogenicity of Erysipelothrix rhusiopathiae
Capsule
Hyaluronidase
Neuramidase/Sialidase
Pathogenic determinant that cleaves N-acetylneuraminic acid, the sialic acid on cell surface
Neuraminidase/Sialidase
Responsible for spread and invasion of E. rhusiopathiae
Hyaluronidase and neuraminidase
Transmission of E. rhusiopathiae
Subcutaneous inoculation through abrasion or puncture wounds
Portal of entry of E. rhusiopathiae
Skin (subcutaneous)
Site of inoculation of E. rhusiopathiae
Fingers and hands (usually like cellulitis)
Primary reservoir of E. rhusiopathiae
Pigs
Most common risk factor for E. rhusiopathiae infection
Occupational exposure
Clinical manifestations of E. rhusiopathiae
Erysipeloid
Diffuse cutaneous eruption with systemic symptoms (Uncommon)
Bacteremia associated with endocarditis
Clinical manifestation of E. rhusiopathiae among pigs
Erysipelas
Cause of Erysipelas in humans
S. pyogenes
Characteristics of erysipeloid
Non-suppurative, purplish erythematous lesions at site of inoculation
Lesions burn and itch
Local cellulitis
This manifestation of E. rhusiopathiae is due to tropism in aortic valve in systemic disease
Bacteremia associated with endocarditis
Laboratory diagnosis of E. rhusiopathiae
Gram stain
Culture
Mouse protection test
Specimens used to culture E. rhusipathiae
deep/full thickness aspirates or biopsies from margin of lesion
True or False.
Swabs are not appropriate because they are localized deep in tissues.
True
Media used to culture E. rhusiopathiae
Heart Infusion (HI) Broth
Blood Agar Plate (BAP)
Erysipelothrix Selective Broth (ESB)
Type of hemolysis caused by E. rhusipathiae
Alpha-hemolytic
CO2 requirement in growing E. rhusiopathiae
5-10% CO2
Capnophile
This differentiates Erysipelothrix from Corynebacterium and Listeria
Production of H2S
Confirmatory test for Erysipelothrix isolates
Mouse protection test
Principle behind mouse protection test
Test mouse is given equine hyperimmune E. rhusiopathiae antiserum for protection
Both control and test mice are inoculated with culture filtrate
After 5-6 days, the mouse without protection dies
General properties of Bacillus
Large, Gram (+) spore-forming rods in chains
General properties of Bacillus anthracis
Straight rods with square/truncate ends
Jointed “bamboo-rod” appearance
Bacillus or B. anthracis
Spores oval, centrally located and do not bulge/distend the cell
Both
Bacillus or B. anthracis
Encapsulated
B. anthracis
Bacillus or B. anthracis.
Spores are formed in culture, by dead/dying animals and not formed by living animals
B. anthracis
Bacillus or B. anthracis.
Non-motile
B. anthracis
Bacillus or B. anthracis.
Usually motile
Bacillus
Bacillus or B. anthracis.
Form rhizoid colonies with peripheral medusa head appearance
Bacillus
Determinants of pathogenicity of B. anthracis
Capsular polypeptide of D-glumatic acid
Anthrax toxin
Only organism to utilize protein (polypeptide of D-glutamic acid) for capsule
B. anthracis
Anti-phagocytic determinant of pathogenicity of B. anthracis
Capsular polypeptide of D-glutamic acid
Plasmid that encode for capsular polypeptide of D-glutamic acid
pXO2
“Invasive stage” of B. anthracis
Capsular polypeptide of D-glutamic acid
Capsular polypeptide of D-glutamic acid is detected by
Mac Fadyean reaction (polychrome methylene blue)
Major virulence factor for B. anthracis
Anthrax toxin
Characteristics of Anthrax toxin
Toxigenic stage
Heat-labile
Trimolecular toxin
Plasmid that encodes for Anthrax toxin
pXO1
Components of the Anthrax toxin
Protective Ag (PA) Edema Factor (EA) Lethal Factor (LA)
Anthrax toxin component that delivers EA and LA to cytosol of cell
PA
Anthrax toxin component that activates the adenylyl cyclase
EA
Anthrax toxin component that is cytotoxic to cells and causes death of target cells
LA
True or False.
None of the three factors is toxic on their own.
True
PA + EA
Edema
PA + LA
Cytotoxicity
PA + EA + LA
Edema and cytotoxicity
Greatest threat from anthrax
The toxin it creates that destroys the victim’s cells even after antibiotics might have killed the bacteria itself.
Principal targets of Anthrax’s Toxin Attack are
Macrophages
Mechanism of macrophage attack during anthrax toxin attack
Anthrax bacteria flood into the bloodstream following
infection
Bacteria produce a toxin comprised of three parts
One part of the toxin, the protective antigen (PA), attaches to a receptor on the membrane, it penetrates the membrane allowing the other toxins to enter
Once inside, the other parts of the toxin kill the cell by disrupting its internal mechanisms
Infectious particle of B. anthracis
Spore
Disease of herbivores, meaning this is acquired from animals and humans are just accidental hosts
Anthrax
True or False.
Anthrax transmission is only exogenous and not contagious because B. anthracis is not part of the normal flora of humans
True
Human cases of anthrax
Agricultural
Industrial
True or False.
There are no cases of person-to-person transmission of B. anthracis.
True
Clinical manifestations of B. anthracis
Cutaneous (Malignant pustule)
Pulmonary (Woolsorter’s disease)
Gastrointestinal (Bowel anthrax)
True or False.
Clinical manifestations of B. anthracis in humans reflects the mode of entry.
True.
Cutaneous - inoculation; most common
Pulmonary - inhalation; most lethal
GI - ingestion; common mode of entry for animals
Epidemiology of Anthrax
Main reservoir of B. anthracis is the soil
Causes death of herbivores upon ingestion of contaminated vegetation
Laboratory diagnosis of B. anthracis
Gram stain Culture and identification Guinea Pig Lethality test Serologic Test PCR
B. anthracis color of capsule after Mac Fadyean Reaction
Blue bacilli surrounded by red capsule
More common stain used to identify B. anthracis
India ink
Specimen used in culturing B. anthracis
Infected/dying animals
Material from pustule, if cutaneous
Media used to culture B. anthracis
BAP
Hemolytic pattern of B. anthracis
Non-hemolytic rhizoid colonies
Test used to demonstrate characteristic appearance of B. anthracis
Guinea Pig Lethality test
Most commonly used serologic test for B. anthracis
ELISA
Serologic tests performed for B. anthracis
Ascoli test
ELISA
IHA
True or False.
Ascoli test lacks specificity for B. anthracis
True
Confirmatory test for B. anthracis
PCR
Treatment for B. anthracis
Penicillin (most commonly used), ciprofloxacin, doxycycline
Prevention of anthrax
Control of disease in animals
Careful handling of infected animals/products
Vaccination
B. anthracis for humans
Killed spore vaccine
Acellular (AVA, AVP)
B. anthracis for animals with disease
Living spore vaccine
Stearne strain
True or False.
Only occupationally at risk individuals are given killed spored vaccine.
True
Anaerobes are unable to utilize O2 as finale electron acceptor because it lacks
Cyctochrome (for aerobic respiration)
Catalase (degrades H2O2)
SOD (degrades superoxide molecule)
Most important enzyme to determine whether the organism can grow in the presence or absence of oxygen
SOD
SOD
Present: _____________
Absent: ______________
Present: Aerobes and facultative anaerobes
Absent: Aerotolerant microbes
Anarobes generate energy solely by
Fermentation
Growth requirements of anaerobes
Low O2 tension (≤ 10% PO2)
Reduced Oxidation-reduction potential (Eh) - expressed in mV
>5% CO2
Enriched medium
Features associated with anaerobic infections
Mostly caused by endogenous opportunistic pathogens
Occur in settings of compromised host defense
Usually not transmissible
Usually polymicrobic
Abscess formation and tissue necrosis
Develops slowly, many are chronic
Putrid odor of infected material/culture
Gas in tissues/loculations
(-) in aerobic cultures
True or False.
Most anaerobic infections are from our own organisms.
True
Putrid odor of infected material is due to
Production of short chain fatty acids as products of metabolism
Reason behind crepitant cellulitis or gas gangrene
Gas in tissues/loculations
Gives you a clue that the infection is cause by an anaerobic culture
(-) in aerobic culture
Laboratory diagnosis for anaerobes
Gram stain
Culture and Identification
Specimens used for anaerobic culture
Sterile specimens Abscess contents, deep wound aspirates Blood, CSF, body fluids Transtracheal aspirates Urine (suprapubic, catheterized)
Media used for anaerobic culture
Should contain reducing agents like sodium thiosulfate, Sodium bisulfate or Sodium sulfide
Egg Yolk Agar (EYA)
Chopped Meat Medium (Chlostridia)
Lake with Kanamycin and Vancomycin Blood Agar (LKVB; Bacteroides)
Employed in LKVB to inhibit the growth of gram-positive organisms
Vancomycin
Employed in LKVB to inhibit gram-negative facultatively anaerobic bacilli
Kanamycin
For furnish culture with reduced O2 and with the proper amount of CO2
Culture systems
Culture systems used to grow anaerobes
GasPak chamber (more common) Anaerobic chamber (more advanced)
Prevention of anaerobic infection
Avoid conditions that reduce Oxidation Reduction Potential in tissues to prevent anaerobiosis in deep tissues
Prevent introduction of anaerobes (from normal flora) into wounds, etc.
Protect against toxin, especially tetanus
Bacteria that is widely distributed in nature, can be found in soil, sewage, and intestinal tracts of humans and other vertebrates
Clostridia
Production of spores in Clostridia
Produced anaerobically, usually distend/bulge the cells
May either be terminal, subterminal, or central
Clostridia or C. perfringens
Motile
Clostridia
Clostridia or C. perfringens
Non-motile
C. perfringens
General properties of C. perfringens
Short, plump Gram (+) rods with squarish ends or “boxcar appearance”
Aerotolerant
Encapsulated
Characteristic of spores in Clostridium perfringens
Oval, central
Do not distend cell
Rarely observed
True or False.
C. perfringens is part of the normal flora of GIT and female genital tract.
True
Five type os C. perfringens
A, B, C, D and E based on production of specific toxins
Most common types of C. perfringens
Type A and Type C (human diseases)
Determinants of pathogenicity of C. perfringens
4 major lethal toxins - alpha, beta, epsilon, iota
Major toxin of C. perfringens
Alpha toxin
Toxin produces by all types of C. perfringens
Alpha toxin
Component of alpha toxin that hydrolyzes phosphorylcholine in cell membrane causing lysis, therefore, death of the cell
Lecithinase C (Phospholipase C)
Toxin produced by B and C
Beta toxin
Toxin produced by types B and D
Epsilon toxin
Toxin produced by type E
Iota toxin
Toxins produced by type B
Alpha, Beta, Epsilon
Toxins produced by type C
Alpha, Beta
Toxins produced by type A
Alpha
Toxins produced by type D
Alpha, Epsilon
Toxin produced by type E
Alpha, Iota
Mode of transmission of C. perfringens
Simple wound contamination
Arises from exogenous and endogenous contamination
Gas formation in soft tissues caused by C. perfringens
Crepitant cellulitis
Highly lethal, necrotizing soft tissue infection of the skeletal muscle
Clostridial myonecrosis/gas gangrene
Characteristics of Clostridial myonecrosis/gas gangrene
Myonecrosis Muscle swelling Severe pain Gas production Sepsis
True or False.
Clostridial myonecrosis/gas gangrene may be cause by C. perfringens, C. novyi, C. septicum.
True
Analogy.
C. perfringens: _______________
C. septicum: _________________
C. perfringens: Traumatic gas gangrene
C. septicum: Non-traumatic gas gangrene
Laboratory Diagnosis of traumatic gas gangrene
Gram stain
Culture
Results of gram staining of traumatic gas gangrene
Predominance of Gram (+) positive rods
Sparse or no WBC
Leukostasis
Spores (rare)
Reason behind sparseness or absence of WBC in traumatic gas gangrene
Alpha toxin hydrolyzes the cell membrane of WBCs
Principle behind leukostasis
Stimulate platelet aggregation so that PMNs cannot penetrate infected tissues
Specimens used to culture C. perfringens
Sterile materials only: tissues, aspirates, deep swabs
Media used to culture C. perfringens
Chopped meat medium
Biochemical methods to chracterize and identify C. perfringens
Double Zone Hemolysis on BAP Opalescence in EYA Nagler reaction Reverse cAMP Stormy fermentation in milk
Double-Zone Hemolysis on BAP is seen as
Inner zone: complete hemolysis
Outer zone: incomplete hemolysis
Principle behind opalescence in EYA
Due to alpha toxin hydrolyzing the lecithin in egg yolk
Confirmatory test for C. perfringens that uses specific antisera that inhibits alpha toxin causing the absence of opalescence
Nagler reaction
Principle behind reverse cAMP
Beta-hemolysis when streaked with S. agalactiae
(+) result when an arrowhead hemolysis is observed towards the test organism
Analogy. Identification of species
Simple cAMP test: _____________
Reverse cAMP test: _____________
Simple cAMP test: identify if culture is S. agalactiae
Reverse cAMP test: identify if culture is C. perfringens
Analogy. Procedure.
Simple cAMP test: _____________
Reverse cAMP test: _____________
Simple cAMP test: Suspected S. agalactiae is streaked on medium and cross-streaked by S. aureus
Reverse cAMP test: Suspected C. perfringens is streaked on medium and cross-streaked with S. agalactiae
Principle behind stormy fermentation in milk
Coagulated milk is disrupted due to gas production of C. perfringens
Treatment for C. perfringens
Cleansing/surgical management of necrotic tissue
Antibiotic therapy: Penicillin, Clindamycin, Metronidazole
Hyperbaric oxygen (adjunct only)
Anti-toxin for α-toxin
Prevention of C. perfringens
Early and adequate wound debridement
Wash with water to avoid anaerobic conditions
Prophylaxis (penicillin)