Agar, Agar, And More Agar!!! Flashcards
Amino Acid (structure)
3 Parts: Carboxyl group, amino group, and “R” side chain
NH3-AA-COOH
I
R
Decarboxylation
Removal of the COOH group (carboxyl group) to COO- (releases CO2)
Deamination
Removal of NH3+ group (amino group)
Lysine
an essential amino acid
Agar slants
Agar which has hardened on a slant
Sensitivity
detects small amounts of the chemical or organism
failure leads to false NEGATIVE
Specificity
discriminates between chemical or organism in question
failure leads to false POSITIVE
Selective media
Suppresses growth of unwanted bacteria and encourages growth of desired microbes
Differential media
Facilitates distinguishing desired microbe from others in a mixed sample
Enterobacteriaceae
Found in human GI tracr
Enterics
General term for enterobacteriaceae
Fecal coliforms
bacterial indicator of sanitary quality of food and water
generally are lactose fermenters
Examples of enteric lactose fermenters
E. coli, Enterobacter, and Klebsiella
these are LESS pathogenic
Examples of non-lactose fermenters
Salmonella, Shigella, Proteus
these are MORE pathogenic
Lactose
disaccharide composed of glucose and galactose
End products of lactose fermentation…
Increase the acidity and make the pH indicator change color
Types of agar used to identify gram (-) microbes
Eosin Methylene Blue (EMB) Agar
Hektoen Enteric (HE/SS) Agar
Eosin-Methylene Blue (EMB) Agar
both selective and differential
selective: eosin and methylene blue dyes suppress growth of gram (+) bacteria
differential: differentiates between vigorous LFs, moderate LFs, and non-LFs
Method for EMB differentiation
Lactose provides the sugar substrate for fermentation (increases acidity of media)
Eosin Y and methylene blue produce a dark purple complex with a green metallic sheen under acidic conditions
Eosin-Methylene Blue (EMB) Agar Results
Gram (+) will not grow on this media
VIGOROUS LFs (like E. coli) grow dark purple with a green metallic sheen
SLOW LFs grow pink colonies
NON-LFs grow colorless colonies
Hektoen Enteric (SS) Agar
both selective (for gram negative) and differential
differentiates among enterics; isolates those that can reduce sulcus to hydrogen sulfide gas (H2S)
How is Hektoen Enteric (SS) Agar Selective
Bile salts inhibit growth of gram (+) bacteria
How is Hektoen Enteric (SS) Agar Differential?
Differentiates between H2S producers and non-producers
Lactose provides sugar substrate for fermentation
Sodium thiosulfate is the sultry source
Ferric ammonium citrate provides a source of iron and reacts with H2S to form a black precipitate
Hektoen Enteric (SS) Agar Results
Gram (+) will not grow on this media
H2S (hydrogen sulfide) producers grow black colonies or blue/green colonies with a black center
Lactose fermenters with NO H2S (like E. coli & Klebsiella pneumoniae) grow yellow to salmon pink colonies
Non-LFs (like Salmonella & Shigella) grow blue/green colonies
Lysine Iron Agar (LIA)
DIFFERENTIAL media that separates gram (-) enteric bacteria upon their ability to decarboxylate and delaminate the amino acid lysine
Contents of Lysine Iron Agar (LIA)
Lysine - sodium thiosulfate
0.1% glucose
Results for Lysine Iron Agar (LIA)
Enterobacteriaceae
Lysine decarboxylase positive (K/K) - purple slant/purple butt
Lysine deaminase positive (R/A) - red slant/yellow butt
Black precipitate - H2S production (K/K, H2S); must be deaminated to observe black precipitate
Lysine Iron Agar (LIA) differentiation
differentiates between Salmonella and Shigella - only Salmonella is an H2S producer (black precipitate)
LIA Interpretation of purple slant/purple butt (K/K)
Lysine deaminase negative; Lysine decarboxylase positive
LIA interpretation of purple slant/yellow butt (K/A)
Lysine deaminase negative; Lysine decarboxylase negative; Glucose fermentation
LIA interpretation of red slant/yellow butt (R/A)
Lysine deaminase POSITIVE; Lysine decarboxylase negative; Glucose fermentation
LIA interpretation of black precipitate (H2S)
Sulfur reduction
Reaction type of LACTOSE FERMENTER
EMB: dark purple with green metallic sheen
SS: pink (or yellow)
Reaction type of of NON-LACTOSE FERMENTER
EMB- colorless
SS- blue-green or colorless