Gram Positive Bacteria Flashcards
3 medically important genus for gram cocci
Staphylococcus
Streptococcus
Gram (-)
Neisseria
Gram positive cocci that is positive in catalase testing
Family: Micrococcaea
Genus: Micrococcus &
Staphylococcus
Gram positive cocci that is negative in catalase testing
Family: Streptococcaceae
Genus: Streptococcus
Gram positive cocci that is oxidation positive for glucose degradation test
Micrococcus (asaccharolytic)
Gram positive cocci that is fermentation/ oxidation positive for glucose degradation test
Staphylococcus (fermentative)
Glucose degradation test
It tests an organism’s ability to ferment the sugar glucose as well as its ability to convert the end product of glycolysis, pyruvic acid into gaseous byproducts.
Catalase test
used to detect the presence of cytochrome oxidase enzyme
-differentiates family micrococcaceae/ staphylococcus (catalase +) from streptococcus(catalase -)
Main reagent in Catalase testing
Hydrogen Peroxide
- Agua oxinada
Procedure for catalase testing
- Add drop of 3% H2O2 solution on glass lside
- then add small amount of bacterial growth from culture medium
Positive reaction for Catalase testing
Bubbling/ Bubbles
Reaction during catalase testing
H2O2 ———> H2O + O2
Test used to differentiate Staphylococcus from Micrococci
Oxidative/ Fermentative test
Another test used to differentiate Staphylococcus from Micrococci
Microdase (modified oxidase) test
(+) blue
Bacteria that is positive in Microdase testing
Micrococcus
Antibiotics that are effective in differentiating Micrococcus and Staphylococcus
Bacitracin (most commonly used)
Furazolidone
Lysostaphin
________ is resistant to bacitracin
Staphylococcus
________ is susceptible/ sensitive to bacitracin
Micrococcus
General characteristics of Staphylococcus species
- gram positive
- catalase positive
- grow best in anaerobic conditions
- facultative anaerobes
- non-motile
- non-sporeforming
5 medically important staphylococcus species
S- saprophyticus H- haemolyticus A- aureus L- lugdunensis E- epidemidis
The most pathogenic Staphylococcus species
Staphlycoccus aureus
MRSA
Methicillin Resistant Staphylococcus aureus
Mode of transmission for Staphylococcus
- Person to person transmission by fomites, air. etc.
- From infected skin lesions such as impetigo, cellulitis, boils (matagtiki)
Staphylococcus specie responsible for 80% suppurative infection in human
Staphylococcus aureus
Dominant site of colonization of Staphylococcus aureus
-site best used to get sample for culture
anterior nares, axilla, perineum, and 10-15% human skin
Principal virulence factor for S. aureus
Coagulase enzyme
Staphylococcus aureus morphology
- gram-positive cocci
- grape like or irregular clusters
- produce lipochrome
- Beta- hemolytic
- haloduric
What causes the golden-yellow pigment of S. aureus on BAP
- production of lipochrome
a naturally occurring fat-soluble pigment
3 major components of S. aureus cell wall
PEPTIDOGLYCAN
- elicits production of interleukin 1 (causes fever) and opsonic antibodies
- provide rigid exoskeleton
- a chemoattractant (attracts polymorphonuclear cells causing inflammations)
- activate complement pathway.
3 major components of S. aureus cell wall
TEICHOIC ACID
- polymer of ribitol phosphate
- regulate the cationic environment of the cell thus control the activity of autolytic enzyme responsible for growth of cell wall
Antiteichoic antibodies are detectable through
Gel diffusion
Antiteichoic antibodies are found generally in patients with __________
- active endocarditis due to S. aureus
3 major components of S. aureus cell wall
PROTEIN A
- major protein component of Staphylococcus
- major antigenic determinant unique to S. aureus
- antiphagocytic by binding thru FC portions of IgG except for IgG 3.
Antiphagocytic
preventing the action of phagocytes or the occurrence of phagocytosis.
PATHOGENICITY
This term refers to the ability of tan organism to induce a local and systemic infection
S. aureus pathogenicity
intracellular
PROTEIN RECEPTORS
- allows organism to adhere to tissue surfaces
S. aureus pathogenicity
intracellular
POLYSACCHARIDE CAPSULE
- protect organism from complement mediated attack of polymorphonuclear (PMN) macrophages, leukocyte and etc.
S. aureus pathogenicity
extracellular
COAGULASE
- this enzyme reacts with blood factor in the plasma to initiate clot formation
- enhance deposition of fibrin around organism to protect from phagocytosis
S. aureus pathogenicity
extracellular
LIPASE
- helps in colonization of organism on oily skin surfaces
- important in survival of organism on sebaceous area of the body
S. aureus pathogenicity
extracellular
HYALURONIDASE
- promote invasion of organism by digesting the intracellular ground substance “glue” (hyaluronic acid) that binds connective tissue in host tissue.
S. aureus pathogenicity
extracellular
STAPHYLOKINASE
- degradation of fibrin clot
(dissolution of clot to spread infection further) - also known as fibrinolycin
S. aureus pathogenicity
extracellular
NUCLEASE (DNAse)
- heat resistant protein
- degrade nucleic acid (DNA)
- lowers viscosity of exodates promoting further motility
S. aureus pathogenicity
HEMOLYSINS - CYTOTOXINS
- lyse rbc by destroying membrane thus inducing tissue damage
- chromosomal mediated
- causes anemia and make iron available for growth of organism
S. aureus pathogenicity
Alpha hemolysin
- most powerful toxin that lyses RBC, platelets, and macrophages thus inducing tissue damage.
- disrupts smooth muscle in blood vessels
S. aureus pathogenicity
Beta hemolysin
- degrades sphingomyelin, and is therefore toxic to rbcs around nerves and many kinds of cells.
- Heat labile
S. aureus pathogenicity
Delta toxin
- disrupts biologic membranes and may have a role in S. aureus diarrheal diseases.
- cytolytic to erythrocytes
S. aureus pathogenicity
Gamma toxin
- a leukocidin that lyses white blood cells
- composed of two proteins designated S and F which may interact with two proteins of Panton- Valentin leukocidin.
PANTON-VALENTINE LEUKOCIDINE
- composed two protein components that kills white blood cells and incapacitate phagocytic line of defense of the host. (necrosis and severe inflammation)
- responsible for various skin, wound, and deep tissue infection.
- encoded on a mobile phage.
S. aureus pathogenicity
ENTEROTOXINS
- heat- stable resistant to the action of gut enzymes
- produced when S. aureus grows in carbohydrate and protein foods.
- responsible for food poisoning caused by S. aureus
Most frequently food poisoning associated enterotoxins
- Enterotoxin A& D
Enterotoxin associated with hospital acquired infection
- Enterotoxin B
Enterotoxin associated with toxic shock syndrome
- Enterotoxin F
S. aureus pathogenicity
EXFOLIATIVE TOXINS
Responsible for:
- Exfoliative dermatitis
- Bullous Impetigo
- Staphylococcal Scarlet fever
S. aureus pathogenicity
TOXIC SHOCK SYNDROME TOXIN
TSST - 1 (SUPERANTIGEN )
- the same with enterotoxin F
- binds to MHC class II mol. yielding t-cell stimulation, which promotes manifestations of TOXIC SHOCK SYNDROME
- associated with: fever, shock, multisystem involvement, including desquamative skin rash.
What causes food poisoning in patients with S. aureus infection?
Enterotoxins produced by the organism not by the organism itself.
Incubation period for food poisoning caused S. aureus
2-6 hrs. followed by abdominal cramps, nausea, vomiting, and diarrhea.
Food poisoning caused by S. aureus
- self-limiting
- rapid recovery which takes place within 24hrs.
Toxic shock syndrome is characterized by
Fever, hypotension, scarlatiniform rashes particularly in the palm and sole which desquamate.
- common among women using vaginal tampoons.
Also referred as pyrogenic Exotoxin C
Toxic shock syndrome
A skin condition caused by exfoliative toxins produced by S. aureus that is common in babies/ neonates.
Staphylococcal Scalded Skin syndrome / Ritter’s Disease
Staphylococcal Scalded Skin Syndrome is characterized by
- diffuse erythematous skin rashes involving the entire body with sloughing off the epidermal layer of the skin.
Skin infections caused by S. aureus
FOLLICULITIS - inflammation in hair follicle
FURUNCLE - spreads to the tissue causing boils
CARBUNCLE - if there is multiple foci and tissue involvement
IMPETIGO - most common manif. in newborn and young children less than 5 yrs. old that is highly communicable.
S. aureus Diagnosis
Gram Stain (presumptive)
Gram positive cocci in clusters
S. aureus Diagnosis
Culture and Isolation
BAP
- beta hemolytic pattern
- golden-yellow pigment
S. aureus Diagnosis
Culture and Isolation
Mannitol Salt Agar
- contains 7.5 % to 10% NaCl
- only S. aureus grow
- pH indicator is phenol red
- turns medium from pink to yellow
Staphylococcus spp. Diagnosis
Culture and Isolation
Phenyl Ethyl Alcohol
Colistin Nalidix Acid agar (CNA)
- inhibits gram negative organism
S. aureus Diagnosis
Culture and Isolation
Chrom Agar
- selective and differential for isolation of MRSA
- (+) mauve/ pink colonies
CONS
coagulase negative staphylococci
S. saprophyticus
S. epidermidis
Coagulase
is an enzyme that converts soluble fibrinogen into soluble fibrin.
The only significant human pathogen that possesses the coagulase enzyme
S. aureus
Two types of coagulase
Cell bound and free coagulase
Two methods for coagulase testing
Method:
Tube - free coagulase (Confirmatory)
slide - cell bound coagulase
Positive reaction for coagulation testing
(+) White fibrin clots
(-) smooth suspension
For tube method in coagulase testing, for how many hrs. will you incubate the tube?
4hrs - check reaction every 30 mins.
-do not extend beyond 4hrs.
Factors that may contribute to coagulase testing of Staphylococcus spp.
- presence of STAPHYLOKINASE enzyme which may lyse the fibrin clot thus may cause a false negative result.
In coagulase testing what type of Plasma is used
Rabbit Plasma/ (Anticoagulated with EDTA)
S. aureus Diagnosis
Biochemical: THERMONUCLEASE TEST
- Staphylococcus contains heat-stable thermonuclease and coagulase negative staphylococcus does not
Positive reaction for Thermonuclease test
(+) Pink zone of clearing at the edge of the well with a darker blue ring at the outer periphery zone.
The only Staphylococci spp. that ferments mannitol
S. aureus