Culture- Growth of microorganism Form or Margin

Streak plate method - common way of

separating bacterial cell on the agar
surface to obtain isolated colonies.
Plate reading- interpretation of primary
culture
Acid fast staining - is a differential
stain to distinguish mycobacteria and
nocardia.
Gram staining- is an empirical method
of differentiating bacterial species into
two large group (gram + and gram -)
based on the chemical and physical
properties of their cell wall.
Hans Christian Gram- Danish scientist
who invented gram staining method.
Fungi- evolutionary advanced form of SIZE
microorganism, as compared to
prokaryotes (prions, viruses, bacteria)  Colonies described as:
 -large
 -medium
Hemolysis:  -small
 Greek word:  -pinpoint
 Size is a visual comparison
Lysis: dissolution or break between its genera or species.
apart
Hemo: pertaining to red
blood cells Density

 a reaction caused especially by

enzymatic or toxin activity of the
bacteria, observed in the media
immediately surrounding or
underneath the colony.
Density colony can be:
 Transparent
 Translucent – allow some light to
pass through the colony
 Opaque – organisms are
concentrated at the center of the
colony described as a bull’s-eye
 colony.(Staphylococci, gram+ &
gram-)

-Dry
-Waxy
Elevation >occasionally, the entire colony will
adheres to the loop.
-is determined by tilting the culture plate
and looking at the side of colony. S. aureus- creamy
It may be: Neisseria spp.- sticky
 Raised Nocardia spp.- brittle, crumbly, and
wrinkled, resembling bread crumbs on a
 Convex
plate.
 Flat
Diphtheroid colonies- dry and waxy
 Umbilicate(depressed center,
Most ᵦ-hemolytic streptococci (except for
concave, an “innie”)
mucoid types)- dry and when push by a
 Umbonate(raised or bulging center, loop, the whole colony remains intact
convex, an “outie”)

Pigment
Color
 Pigment production is an inherent
 Colonies maybe: characteristics of a specific
organism defined generally to the
 White: Coagulase-negative colony.
Staphylococci
Odor
 Gray: Enterococcus spp.
 Odor should be determined when
 Yellow or off white: Micrococcus the lid of the culture plate is
species and Neisseria species removed and its odor dissipates
 Buff: “Diphtheroids” into the surrounding environment.

CONSISTENCY  NEVER INHALE directly from the

plate
 Determined by touching the
colony with a sterile loop. Examples of microorganisms that
produces distinctive odor includes:
 It may be:
 S. aureus – old sock (stocking
-Brittle (splinters) that has been worn continuosly
-Creamy (butyrous) for a few days without washing);
 this odor evident when growing Ziehl-Neelsen carbol fuchsin- primary
on mannitol salt agar. stain (red)
 P. aeruginosa – fruit or grape – Pos-red neg- red
like.
Acid alcohol (3% HCI + 95% Ethanol)
 P. mirabilis – putrid. -decolorizer
 Haemophilis spp. – musty Pos- red neg- colorless
basement, “mousy” or “mouse
Methylene blue – counter stain
nest” smell.
Pos- red neg- blue
 Nocardia spp. – freshly plowed
field.
Fungal morphology (act 9)
Reagents:
Simple staining (act 5):
0.9%NSS
Methylene blue- only reagent
Potassium hydroxide

Gram staining (act 7)

Reagent:
Crystal violet –primary stain
Pos- violet neg- purple

Iodine
Pos- violet/purple neg- purple

95% ethyl alcohol- decolorizer

Pos- purple/purple neg- colorless

Safranin- counter stain

Pos-purple neg-pink/red

Acid fast (act 8)

Sputum