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Microbiology. Medically Important Staph and Strep Labs 24 - 26. MSA plates. Media pH Phenol Red Mannitol Pathogenic ferment mannitol and acids change phenol red to yellow Nonpathogenic grow no color change 7.5% NaCl
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Microbiology Medically Important Staph and Strep Labs 24 - 26
MSA plates • Media • pH Phenol Red • Mannitol • Pathogenic ferment mannitol and acids change phenol red to yellow • Nonpathogenic grow no color change • 7.5% NaCl • Selective medium for staphylococci since most other pathogenic bacteria cannot survive in high salt
Coagulase Production • Used to differentiate S. Aureus from other gram positive cocci • S. aureus is an opportunistic pathogen known to be highly resistant due to production of coagulase • Coagulase works in conjunction with normal serum components to form protective fibrin barriers around bacterial cells which shields them from phagocytosis and other types of immune attacks
Coagulase Production • Coagulase enzymes occur in two forms • Bound • Attached to cell wall reacts directly with fibrinogen in plasma causing fibrinogen to precipitates which causes clumping • Free • Extracellular enzyme that reacts with plasma component coagulase reacting factor (CRF) which causes normal clotting mechanisms to begins
Coagulase Production • In our test the yellow colonies from the MSA plate are presumed pathogens • However, virulence of the organism is not established until coagulase test is done • Rabbit plasma is treated with sodium citrate a natural anticoagulant • If coagulase is present then it inactivates the sodium citrate and allows the plasma to clot
Coagulase Production • Substrate: Sodium citrate • Enzmye: Coagulase • pH indicator/reagents: none • Pos: Clotting • Neg: Liquid
Hemolysin Production • Blood agar is used for isolation and cultivative of many types of fastidious bacteria • Used to differentiate bacteria within the genera Streptococcus, Enterococcus, and Aerococcus
Hemolysin Reaction • Several species of gram positive cocci produce exotoxins known as hemolysins • Hemolysins able to destroy RBC’s and hemoglobin (except gamma hemolysin) • Beta – complete destruction of RBC’s and hemoglobin results in clearing • Alpha – partial destruction of RBC’s heme group not used greening ting seen • Gamma – non RBC hemolysis only plasma protein appears as simple growth no change to medium
Alpha (Left) and Gamma (Right) Hemolysis Greenish ting Heme group not broken down agar remains red color