All O SAL AGAR

- differential media ⇒ used to visually distinguish microorganisms from one another

- selective media ⇒ used to grow/isolate specific types of microorganisms suppressing the growth of other microorganisms
- mannitol salt agar (USA) contains the carbohydrates mannitol, 7.5% sodium chloride (NaCl), and the pH indicator phenol red
- components
↳ phenol red indicate ⇒ yellow below pH = 6.8
⇒ red at pH = 7.4 - 8.4

⇒ pink at pH = 8.4 and above

↳ mannitol ⇒ gives substrate for fermentation and makes medium differential

↳ sodium chloride ⇒ makes medium selective since its concentration is high enough to dehydrate/kill most bacteria
• bacteria

↳ staphylococci thrive on this medium due to its adaptation to salty habitats such as human skin

↳ phenol red ⇒ indicates whether fermentation with an acid has taken place by changing color as the pH changes

↳ most staphylococci are able to grow on MSA, but do NOT ferment mannitol, showing growth as pink or red and the medium stays unchanged

↳ Staphylococcus aureus ferments mannitol, producing acids and lowers medium pH ⇒ bright yellow colonies form
• application ⇒ mannitol salt aga.r used for isolation and differentiation of Staphylococcus aureus from other StaphylococcuS species

BLOOD AGAR
• blood agar separates Gram + cocci that produces exotoxins called hemolysins

- hemolysins ⇒ lipids/proteins that break down red blood cells

• blood agar is made of 5% of blood (sheep blood) in a tryptic soy agar base

blood agar allows differentiation of bacteria based on their ability to hemolyze red blood cells

3 types of hemolysis

↳ beta-hemolysis ⇒ complete destruction of red blood cells and hemoglobin, resulting in a clearing of the medium around the colonies

↳ alpha-hemolysis ⇒ partial destruction of red blood cells and produces an olive-greenish discoloration of the agar around the colonies
in reflected light → converting heme in hemoglobin to methelglobin that cannot bind oxygen, appearing as green
↳ gamma-hemolysis ⇒ nonhemolysis, appearing as simple growth with no change to the medium

- hemolysins produced by streptococci ⇒ streptolysins

. 2 forms ⇒ Type O and Type S
↳ streptolysin 0 ⇒ oxygen sensitive, grows best under anaerobic conditions (in agar)
↳ streptolysin S ⇒ oxygen stable, grows in aerobic conditions surface of agar plate)

- streak stab technique ⇒ method to provide environment favorable for streptolySins

↳ blood agar plate streaked for isolation and then stabbed with a loop
↳ alpha-hemolysis on the surface but beta-hemolysis in the stab

BAA RACH SUSCEPT/BHi T St

. bacitracin ⇒ made by Bacillus licheniformis is a powerful peptide antibiotic that inhibits bacterial cell wall synthesis by

interfering with peptidoglycan transport across the cytoplasmic membrane → effective only to bacteria with cell walls
that are in the process of growing
• novobiocin ⇒ antibiotic produced by Streptomyces nivens →
interferes with ATPase activity associated with DNA gyrase
(enzyme needed for DNA replication)

- optochin ⇒ antibiotic derived from quinine that disrupts ATP synthase activity - reduced ATP production in susceptible bacteria
- bacitracin test is used to differentiate and identify ⇒ beta-hemolytic group A streptococci (Streptococcus pyogenes - bacitracin susceptible)
from other beta-hemolytic streptococci (bacitracin resistant)