PHMicroPara (Lab) | Experiment 4: PREPARATION OF CULTURE MEDIA

Culture Media

are nutrient preparations used for growing microorganisms.

Culture Media

are nutrient materials or substances used to grow in vitro.

Culture Media

• Obtained from different algae.

• No nutritive value.

• Not affected by growth of bacteria.

• Resistant to action of microorganism.

Culture

term used to microorganisms that are cultivated in the lab

Medium

Is the term given to the combination of ingredients that will support the growth and cultivation of microorganisms by providing all the essential nutrients required for the growth

Nutrient Media

TYPES OF CULTURE MEDIA: basic growth.

Enriched Media

TYPES OF CULTURE MEDIA: contains special nutrients.

Selective Media

TYPES OF CULTURE MEDIA: favors certain microbes.

Differential Media

TYPES OF CULTURE MEDIA: distinguishes organisms.

Transport Media

TYPES OF CULTURE MEDIA: preserves microbes during transfer.

Accurately

General Steps in Preparation:

1. Weighing ingredients?

Distilled Water

General Steps in Preparation:

2. Dissolving components in?

pH

General Steps in Preparation:

3. Adjusting ___ to required level.

4. Dispensing

General Steps in Preparation:

4. ________ into appropriate containers

Autoclaving

General Steps in Preparation:

5. Sterilization by?

121

STERILIZATION: Most culture media are sterilized by autoclaving at ___°C for 15 minutes

15

STERILIZATION: Most culture media are sterilized by autoclaving at 121°C for ____ minutes

Heat-Labile Components

STERILIZATION: Some media require filtration (__________).

45–50

POURING MEDIA: After sterilization, allow media to cool to ___-___°C.

Aseptically

POURING MEDIA: Pour _______ into Petri dishes or tubes.

Air Bubbles and Contamination

POURING MEDIA: Avoid these

Label and Store

POURING MEDIA: _____ and ___ appropriately

2–8

STORAGE AND QUALITY CONTROL: Store prepared media at ___ - ___ °C.

Contamination

STORAGE AND QUALITY CONTROL: Check for _________ before use.

pH, Color and Clarity

STORAGE AND QUALITY CONTROL: Ensure consistency in _____, ______ and __________

Discard

STORAGE AND QUALITY CONTROL: _______ expired or contaminated media.

Clinical Diagnosis

PURPOSES OF BACTERIAL CULTURE: Aid in?

Morphology and Identification

PURPOSES OF BACTERIAL CULTURE: Study bacterial _________ and ________.

Serological Tests and Vaccine Development

PURPOSES OF BACTERIAL CULTURE: Produce antigens for?

Genetic Research and Cell Manipulation

PURPOSES OF BACTERIAL CULTURE: Facilitate ___________ and _________ in vitro

Isolation and Separation

PURPOSES OF BACTERIAL CULTURE: Use of solid media for _________ and __________ of bacteria in mixed samples

Culture Media

Microbiological culture involves growing microorganisms.

Culture Media

Specialized media are necessary for cultivating specific microorganisms or cell culture

Culture Media

Used for cultivating bacteria

Culture Media

Functions: Increase bacterial population.

Culture Media

Functions: Selecting specific bacteria.

Culture Media

Functions: Differentiate various bacterial types.

Artificial Medium

NEED FOR CULTURE MEDIA: Growing the organism in an ________________ is necessary to obtain a culture.

Nutrients and Mineral Salts

BASIC REQUIREMENTS OF CULTURE MEDIA requires?

Energy Source, Carbon Source and Nitrogen Source

BASIC REQUIREMENTS OF CULTURE MEDIA: NUTRIENTS consists of?

Energy Source

BASIC REQUIREMENTS OF CULTURE MEDIA | NUTRIENTS: Phototrophs and chemototrops are used as?

Phototrophs

BASIC REQUIREMENTS OF CULTURE MEDIA | NUTRIENTS: light (e.g., Rhodospirillum rubrum)

Rhodospirillum rubrum

This bacteria is a phototroph

Chemotrophs

BASIC REQUIREMENTS OF CULTURE MEDIA | NUTRIENTS: chemical energy (e.g., Escherichia coli)

Escherichia coli

This bacteria is a chemotroph

CO2 or Organic Compounds

BASIC REQUIREMENTS OF CULTURE MEDIA | NUTRIENTS: Carbon Source consists of?

Atmospheric Nitrogen, Nitrate or Nitrite

BASIC REQUIREMENTS OF CULTURE MEDIA | NUTRIENTS: Nitrogen Source consists of?

Mineral Salts

BASIC REQUIREMENTS OF CULTURE MEDIA: Sulfates, phosphates, chlorides, and carbonates of potassium (K), magnesium (Mg), and calcium (Ca)

7.2-7.4

BASIC REQUIREMENTS OF CULTURE MEDIA | MINERAL SALTS: Optimal pH is? (___-___)

Nutrient agar

Example of Nutrient Media

Bllod agar, Chocolate agar

Example of Enriched Media

McConkey Agar

Example of Selective Media

McConkey Agar

Example of Differential Media

Nutrient Agar

Purpose: General growth of non-fastidious bacteria

Enriched Media

Purpose: Growth of fastidious organism

Selective Media

Purpose: Allows Gram-negative growth, inhibits others

Differential Media

Purpose: Differentiate lactose fermenters from non-fermenter

Mueller–Hinton Agar

MHA stands for?

Mueller–Hinton Agar

A nutrient-rich, non-selective medium commonly used in microbiology.

Mueller–Hinton Agar

Main Use: Standard medium for antibiotic susceptibility testing (e.g., Kirby–Bauer disk diffusion test).

Antibiotic Susceptibility Testing

Mueller–Hinton Agar is a standard medium for?

Mueller–Hinton Agar

Composition: Contains beef infusion, casein hydrolysate, starch, and agar

Beef Infusion, Casein Hydrolysate, Starch and Agar

Mueller–Hinton Agar contains?

Mueller–Hinton Agar

Classification: Special Purpose Media

Escherichia coli

Nutrients: Arginine

Specimen: ?

Gonococci

Nutrients: Glutathione

Specimen: ?

Mycoplasma

Nutrients: Cholesterol

Specimen: ?

Atypical Mycobacteria

Nutrients: Aryl Sulphate, Amide

Specimen: ?

Mycoplasma hominis

Nutrients: Glycerol

Specimen: ?

Rickettsia

Nutrients: Sulfonamides

Specimen: ?

Salmonella typhi

Nutrients: Tryptophan

Specimen: ?

Listeria monocytogenes

Nutrients: L-Cysteine

Specimen: ?

Vibrio parahaemolyticus

Nutrients: Sodium Chloride

Specimen: ?

H. influenza

Nutrients: Factor X & V

Specimen: ?

Liquid, Semi-Solid and Solid

Types Based on Consistency

Aerobic and Anaerobic

Types Based on Classification

Simple, Complex and Synthetic, Special Media

Types Based on Type of Agar

Nutrient Broth

It is also used to test for indole formation.

Nutrient Broth

Culturing organisms to demonstrate motility.

Enriched Medium

What type of medium is Blood Agar?

Blood Agar

Composition: Sterile nutrient agar combined with defibrinated sheep blood

Blood Agar

Uses:

• Routine culturing

• Commonly used in medical bacteriology

• Serves as an indicator medium to detect hemolytic activity of bacteria, such as Streptococcus pyogenes

Streptococcus pyogenes

BLOOD AGAR: Serves as an indicator medium to detect hemolytic activity of bacteria, such as?

Neisseria, Haemophilus influenzae and Pneumococcus

Chocolate Agar is used to cultivate?

Nutrient Broth, Peptone Broth, Glucose Broth and Tryptic Soy Broth

Types of Broth Media

Nutrient Broth

TYPES OF BROTH MEDIA: general-purpose medium for non-fastidious bacteria

Peptone Broth

TYPES OF BROTH MEDIA: simple broth, mainly for growth and fermentation tests

Glucose Broth

TYPES OF BROTH MEDIA: contains glucose as a carbohydrate source for bacterial growth

Tryptic Soy Broth

TYPES OF BROTH MEDIA: enriched general purpose broth for a wide range of bacteria