MODULE 4.1 Flashcards by Ateeya Cali

increase in the number of cells or microbial population rather than in the size of individual cells

Microbial Growth

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To increase the population of
microorganisms, their _____ and
_____ requirements must be provided.

nutritional and physical

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Factors Affecting Microbial Growth

biochemical factors (nutrition)
physical factors
generation time

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it is supplying cells with chemical tools (nutrients) they need to make monomers of macromolecules that mainly comprise microbial cells

Microbial Nutrition

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made up of chemical elements
extracellular substances that provide the microbial cell with materials

Nutrients

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Nutrients are extracellular substances that provide the microbial cell
with materials to (2)

Build protoplasm

Generate energy

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Biochemical factors (nutrition)

Macronutrients

Micronutrients

Vitamins

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Macronutrients

Protein, lipid, polysaccharide, LPS DNA, and RNA

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Micronutrients

C, H, O, N, P, S, K, Na, Ca, Mg, Fe

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Vitamins

B1 (Thiamine), B2 (Riboflavin), B6 , B12 (Cobalamine), K, Biotin, Folic acid

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any nutrient material prepared/used for the growth and cultivation of microorganisms in the laboratory

Culture Medium

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What are Culture Media for?

for the growth and maintenance of microbial cultures
to favor the production of particular compounds
to study microbial action on some constituents of the medium

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Types of Culture Media (physical composition)

Liquid
Semi-solid
Solid

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Types of Culture Media (chemical composition)

Synthetic or chemically-defined

Complex

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Types of Culture Media (principal purpose, function, or application)

General Purpose
Differential
Selective
Enrichment
Assay

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Nutrient Agar
Tryptic Soy Agar
Brain Heart Infusion Agar

General purpose culture media

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Eosin Methylene Blue
Mannitol Salt Agar
Blood agar (Streptococcus species)
Mac-Conkey agar (E. coli and lactose fermenters)

Differential

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contains general nutrients and 5% sheep blood.

It is useful for cultivating fastidious
organisms and for determining the hemolytic capabilities of an organism.

Blood agar

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Some bacteria produce different types of exoenzymes that lyse red blood cells
and degrade hemoglobin;

hemolysins

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different types of hemolysins - result in blood agar

beta hemolysis - clear zone
Alpha-hemolysin - greenish color
gamma hemolysis - no clearing

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Thayer-Martin agar (Neisseria gonorrhoeae)

NA with penicillin (Gram-negative bacteria)

Thiosulfate Citrate Bile Salts Sucrose (TCBS) Agar;

Selective culture medium

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Cellulose agar (for cellulolytic microorganisms)

Petroleum broth (for petroleum-utilizers)

Blood agar (hemolytic bacteria)

Enrichment culture medium

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fermentation media
TSI agar
Vitamin B12 assay medium

Assay culture medium

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Other Types of Culture Media

Indicator Medium
Sugar Medium
Transport Medium
Biochemical Reaction Medium
Aerobic Media and Anaerobic Media

medium with no solidifying agent, used for inoculum preparation, fermentation test Nutrient Broth; Lactose Broth

liquid culture media

medium with 0.1- 0.5% solidifying agent, used for motility test, Sulfur Indole Motility (SIM) Medium

semi-solid culture media

medium with 1.5-2.0% solidifying agent, used for Colony morphology observation, hemolysis and pigmentation characterization Nutrient Agar; Blood Agar

solid culture media

types of agar solid culture medium

agar plate agar slate agar deep

complex polysaccharide (usually derived from red algae) used as solidifying agent for culture media in Petri plates, slants, and deeps

Agar

T or F. Agar is affected by growth of bacteria and has nutritive value.

F. Agar has no nutritive value; generally not metabolized by microbes and is not affected by growth of bacteria.

Agar * Liquefies at _____ * Solidifies at _____

100°C ~40°C

medium where all components are chemically-defined (precise nutrient composition and amounts) prepared from pure chemicals uses: important for genetic and specific or precise studies Glucose, Inorganic Salt, Phosphate

Synthetic or chemically- defined medium

disadvantages of synthetic medium (3)

– preparation is time-consuming – microorganisms grow relatively slow – prepared only for microorganisms with known nutritional requirements

medium where not all components are chemically-defined made from extracts from natural materials (beef, blood, casein, yeast, soybeans) use: routine purposes Nutrient Agar, yeast extract

Complex medium

advantages of complex medium

– easy to prepare – support rapid growth of most microorganisms

medium that can support most or almost all types of species

general purpose

medium that can distinguish visually one type of bacterium from another exploits the ability of a particular microorganism to change the appearance of the medium with special reagents like pH indicators and dyes

differential

medium that allows growth of a specific type of microorganism only contains selective agents/additives/toxic chemicals (salts, dyes, antibiotics, and other inhibitors) extreme pH value or unusual carbon source to favor growth of a particular organism

selective

selective medium used for Neisseria gonorrhoeae

Thayer-Martin Selective Agar

medium that is used to increase the number of microorganisms with unusual physiological characteristics; contains special nutrients for microorganisms of interest contains special nutrients for microbes of interest and inhibitory substances to suppress unwanted microorganisms

enrichment

Enrichment medium for hemolytic bacteria

Blood Agar

medium that is used for assay of vitamins, amino acids, antibiotics; may be used for qualitative or quantitative production of a compound by a microorganism

assay

medium contains an indicator which changes its color when a bacterium grows in them Blood agar, Mac Conkey’s medium

indicator medium

medium containing any fermentable substances consists of 1% of the sugar in peptone water

Sugar Medium

a small tube in sugar medium for the detection of gas by bacteria

Durham tube

is a differential and sugar medium that contains 1% lactose, 1% sucrose, 0.1% glucose, ferrous sulfate , pH indicator PHENOL RED Used to determine the ability to: 1. Reduce sulfur to H2S 2. Lactose &/or Sucrose fermentation 3. Glucose fermentation and Gas production

Triple Sugar Iron Test (TSI)

medium used for transporting samples (prevent microbial proliferation; maintain viability of microorganisms) Stuart’s medium, Cary-Blair, Amies

Transport medium

medium that is recommended for the preservation and transportation of Neisseria species and other fastidious organisms from the clinic to laboratory.

Transport Medium Stuart

medium used to provide additional information for the identification of the bacterium Triple sugar iron agar (sugar fermentation); SIM Medium (Indole test); Citrate utilization; Christensens urease medium (Urease test)

Biochemical Reaction Medium

medium for SUGAR FERMENTATION

Triple sugar iron agar * Lactose fermenters - E.coli, Klebsiella * Non Lactose fermenters - Salmonella, Shigella * H2S - Proteus

Used to detect indole production by the organism. Motility Test Kovac's Reagent

SIM Medium (Indole test, motility test) (+) pink ring - E.coli (-) yellow ring - Klebsiella, Salmonella

medium used in CITRATE UTILIZATION TEST Test used to detect the ability of certain bacteria to utilize citrate as the sole source of carbon. Sodium citrate and bromothymol blue as the indicator

Simmon’s Citrate medium (+) blue color: Klebsiella (-) green color: E.coli

medium used in UREASE TEST test is used to detect organisms that produce urease.

Christensen’s urease medium (+) pink color: Proteus, Klebsiella (-) yellow color: E.coli, Salmonella

medium that is used to grow aerobes - Mannitol salt agar

Aerobic medium

medium that are used to grow anaerobes - Robertson’s cooked meat medium - Thioglycolate medium

Anaerobic medium

anaerobic medium that uses cooked meat particles as a reducing agent that absorbs O. most species produce gas in this medium

Robertson’s cooked meat medium Saccharolytic species turn meat pink Proteolytic species turn meat black with foul smell

Anaerobes that attack meat and turns meat into pink in Robertson’s cooked meat medium

Saccharolytic species

Anaerobes that attack carbohydrates in meat and turns meat into black with foul smell Robertson’s cooked meat medium

Proteolytic species