BACTERAIL CULTURE METHODS

Prajith.V PharmD @Sri Ramakrishna Institute of Paramedical Sciences Prajithkanna@gmail.com

BACTERAIL NUTRITONS AND THE DESIGN OF CULTURE MEDIA

. Based on bacterial Metabolism*

. Culture pH

. Culture Oxidation-reduction Potential

. Gaseous Requirements . Oxygen, Carbon dioxide and other gases

OXYGEN CONCENTRATIONS

Aerobs Anaerobs (do not require Oxygen) Obligate Anaerobs (die in the presence of Oxygne) Facultative anaerobs (E.coli) Microaerophilic Bacterial

PURPOSE OF CULTURINGIsolationProperties of bacteriaTo create antigens for laboratory useTyping with Bacteriophages and Bacteriocins

SusceptibilityTo test for Antibiotic sensitivityEstimate viable countsMaintain stock cultures

METHODS OF ISOLATIONS OF PURE CULTURE WITH

SURFACE PLATING ENRICHMENT MEDIUM

SELECTIVE MEDIUM

INDICATOR MEDIUM

TEMPERATURE AND BACTERIAL GROWTH

THE REQUIREMENTS FOR GROWTH PHYSICAL REQUIREMENTS

METHODS TO ISOLATE THE BACTERIAStreak Culture

Stroke Stab Pour plate Liquid culture

Special Methods for anaerobic cultures

STREAK CULTURE

How do you count bacteria ?Comparing Bacteria Counts of Water Samples. Put a drop of water of well water on a sterile

plate of TGY or other agar. Spread it uniformly over the surface of the

agar with any non-absorbent sterile tool. ... Incubate the plate at 30C or room temperature (r.t.). Examine the plate as often as you like. ... Count the colonies.

What is a cell counter?A counting chamber,(also known as hemocytometer),

is a microscope slide that is especially designed to enable cell counting. The slide has a sink in its middle; the area of the sink is marked with a grid. A drop of a cell culture is placed in the sink

What is a serial Dilution in Microbiology?We can dilute once, then dilute this dilution, only to dilute this dilution, and so on until we get to the appropriate concentration of cells. This is called a serial dilution. A serial dilution is a series of sequential dilutions used to reduce a dense culture of cells to a more usable concentration.

How do you calculate viable cell count? Take the average cell count from each of the sets of 16 corner squares. Multiply by 10,000 (104). Multiply by 5 to correct for the 1:5 dilution from the Trypan Blue addition.

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