Module 4 - Culture Media (Clin Bacteriology Laboratory)

Culture

growth of an organism in a culture media

Culture media

artificial laboratory environment for the growth of an organism

Pure culture

only contains one species of bacteria

Mixed Culture

contains multiple bacteria of interest

stock culture

designed to maintain bacteria viable

contaminated culture

culture containing a contaminant

culture media

anything that possesses nutrition and environmental requirements (temperature and pH) necessary for bacterial growth

Chemically-defined

Synthetic Media is also known as?

Synthetic media

Contains exact chemical composition and amount of all components are known?

Mannitol Salt Agar

What is an example of synthetic media?

Non-synthetic

Contains nutrients in which the exact components are or quantity is unknown

Non-chemically defined or Complex

Non-synthetic media is also known as?

Nutrient Agar

An example of non-synthetic media

Peptone

Nutrient agar contains ____ wherein the chemical composition is unknown.

Tissue culture media

Culture media that is composed of live cells from animals or humans

Tissue culture media

This culture media is rarely used for the culture of bacteria, and is mainly used in the culture of viruses

Vero Cell Line

Composed of kidney cells of African Green Monkey

Mccoy Cell Line

Composed of Mouse Cell Line

Chicken Embryo

Fertilized Chicken Egg

A549 Cells

Composed of Human Lung Carcinoma

HELA Cell Line

Composed from Human Cervical Carcinoma

Hep-2 Cell Line

Composed of Human Epithelial Cell Carcinoma of the Larynx

Agar or Agarose

Solidifying agent used in culture media is

Liquid

Which type of culture media is commonly used for rapid culture?

Broth

Liquid media is also known as?

Trypticase Soy Broth, Nutrient broth, Brain Heart Infusion

Examples of Liquid Culture media

Turbidity

Positive growth of bacteria in a liquid broth is indicated by the presence of?

0.5-1.0% agar

How much agar is present in Semi-solid media?

Semi solid

Which media is useful for studying motility

SIM (Sulfide Indole Motility) Media

Media used in demonstrating motility

Spreading of bacteria from the site of innoculation

This indicates positive in motility in a semi-solid media

Bacteria is steady and still from the site of innoculation

This indicates negative motility in a semi-solid media

2-3% agar

Amount of agar present in a solid media

False

TRUE OR FALSE. Most culture media that are used are semi-solid

Solid Liquefiable

These are the commercially available culture media where it is bottled and powdered form.

Dissolve in H2O → Sterilize → Place in petri dish → Allow to solidify

Process of preparing Solid Liquefiable media

Solid Non-Liquefiable

Does not melt

Rice media, potato slices, cooked meat

Examples of Solid Non-Liquefiable Media

Biphasic

There are 2 physical states in one culture media

Castaneda bottle

Example of Biphasic Media

Blood Culture

Castaneda bottle is used for?

Plated Media

solidified in a petri dish

100 mm

What is the Standard size of Petri Dish

15-20mL

How much culture media is used in a plated media?

150mm

Size of petri dish used for AST

Liquid, semi-solid, solid

Tubed media can be in which physical states?

Liquid, Upright

Physical state/s and formation of Broth Media

Mixing with Inoculating loop

Manner of inoculation for broth media

Solid/Semi-Solid

Physical state/s of butt media

Stab

Manner of inoculation for semi-solid media?

Always solid

Physical state/s for slant

Streak

Manner of inoculation for slant

Always solid

Butt slant is always present in which physical state?

Stab in the butt area and streak in the slant area

Manner of inoculation for butt-slant

Bottled media

Used for Blood Culture

General Purpose Media

Supports the growth of non-fastidious microorganisms.

General Isolation Media/Nutritive Media

General Purpose Media is also known as?

General Purpose Media

Used as base media in preparation of other culture media

Nutrient agar/broth, Tryptic soy agar/broth

Examples of General Purpose Media

Enrichment Culture Media

Enhances growth of a particular bacteria

Alkaline Peptone Water (Broth)

Enrichment Culture Media for Vibrio species

Selenite F Broth

Enrichment Culture Media for Salmonella spp

Normal intestinal flora

Selenite F Broth inhibits the growth of?

Thioglycolate Broth

Enrichment Culture Media for anaerobic bacteria

mineral oil

Thioglycolate Broth adds ______ to prevent the exposure of the culture media to air

Gram Negative broth

Enrichment culture media for Salmonella and Shigella

Normal intestinal flora

Gram Negative broth inhibits the growth of?

Todd Hewitt broth

Enrichment media for Beta-hemolytic streptococcus, S. pneumoniae

Colistin and Nalidixic Acid

Lim broth is a Todd-Hewitt Broth with added?

Streptococcus agalactiae

Lim broth is an enrichment media for?

vaginal swab specimen

Lim broth is an enrichment culture media for S. agalactiae on which specimen?

Neonatal meningitis

Pregnant women are recommended to be screened for S. agalactiae because it is a common cause of?

Enriched Culture Media

Contains basic nutritional requirement to support the growth of non-fastidious organisms with additives, enriching substances, or supplements to support the growth of fastidious microorganisms

Blood Agar plate, Chocolate Agar plate

Examples of enriched culture media

Dissolve BAP in H2O → Sterilize (Autoclaving) → Cool it down → Add the blood

Process of preparation of Blood Agar Plate?

Dissolved CAP in H2O → Add the blood → Sterilize (Autoclaving)

Process of preparation of Chocolate Agar Plate?

Due to hemolyzed RBCs and denatured hemoglobin

Why does chocolate agar plate turns chocolate brown color?

X Factor

Which factor/s is/are present in BAP?

X and V Factor

Which factor/s is/are present in CAP?

Hemin

X factor is?

Nicotinamide Adenine Dinucleotide

V factor is?

Selective Media

Favor the growth of organism of interest

Cystine Tellurite Blood Agar

Selective media for C. diphtheriae

Potassium tellurite

Inhibitor present in the selective media for C. diphtheriae

Malachite green

Inhibitor present in the selective media for M. tuberculosis

Gram negative bacteria

Malachite green is an inhibitor of LJ media, it inhibits?

Normal flora

Potassium tellurite is an inhibitor Cystine Tellurite Blood Agar, it inhibits?

Lowenstein Jensen Media

Selective media for M. tuberculosis

Thayer Martin Media, Modified Thayer Martin Media, Martin Lewis Media, New York City Media, and GC-LECT

Selective media/s for N. gonorrhoeae

Colistin

Inhibitor for gram negative bacteria of the selective media for Neisseria gonorrhoeae (Thayer Martin Media)

Vancomycin

Inhibitor for gram positive bacteria of the selective media for Neisseria gonorrhoeae (Thayer Martin Media)

Nystatin

Fungal inhibitor of the selective media for Neisseria gonorrhoeae (Thayer Martin Media)

Colistin

Inhibitor for gram negative bacteria of the selective media for Neisseria gonorrhoeae (Modified Thayer Martin Media)

Vancomycin

Inhibitor for gram positive bacteria of the selective media for Neisseria gonorrhoeae (Modified Thayer Martin Media)

Nystatin

Fungal inhibitor of the selective media for Neisseria gonorrhoeae (Modified Thayer Martin Media)

Trimethoprim lactate

Inhibitor for swarming organisms of the selective media for Neisseria gonorrhoeae (Modified Thayer Martin Media)

Proteus

What do we call the swarming organisms?

Trimethoprim lactate

Inhibitor for swarming organisms of the selective media for Neisseria gonorrhoeae (Martin Lewis Media)

Anicomycin

Fungal inhibitor of the selective media for Neisseria gonorrhoeae (Martin Lewis Media)