Selective and Differential Media

CNA-blood agar

Colastin Naladixic Acid plus sheep's blood; selects for gram(+) bacteria, inhibits gram(-) bacterial growth

CNA-blood selective ingredient

Colastin Naladixic Acid

CNA differential ingredient

Sheep red blood cells

alpha hemolysis

partial hemolysis of the sheep red blood cells. produces andolive-green color in the blood

beta hemolysis

clearing of the media as the enzymes in the bacteria degrade the red blood cells.

gamma hemolysis

no hemolysis of the red blood cells

MacConkey media selective ingredient

Analine dyes such as crystal violet; bile salts

MacConkey differential ingredient

lactose

Maconkey media

selects for gram(-) organisms, inhibits gram(+) bacteria

MacConkey pH indicator

neutral red; produces fuschia bacteria if the bacteria can ferment the lactose

MSA

Manitol salt agar; selects for gram(+), salt tolerant bacteria. Inhibits bacteria that can not tolerate salt

MSA selective ingredient

salt

MSA differential ingredient

manitol

MSA pH indicator

Phenol red. This pH indicator turns yellow at low pH, fuschia at high pH, and is red at neutral pH

TSI media

Triple sugar iron media differentiates enterobacteria according to how they ferment different sugars, whether or not they produce gas and whether or not they produce a black percipitate

TSI media sugars

TSI media contains glucose, lactose and sucrose. A bacteria will utilize the glucose first, and the dissacharides second.

TSI pH indicator

phenol red

TSI sugar fermentation results (K/A)

Slant/butt can be red/yellow (K/A) indicating the bacteria can ferment glucose only, not the dissacharides

TSI sugar fermentation results (A/A)

Yellow slant and yellow butt indicate fermentation of the glucose and one or both of the dissacharides (lactose and sucrose)

TSI gas results

bubbles of carbon dioxide gas can show up next to the glass or as cracks in the media

TSI hydrogen sulfide production

A black color indicates hydrogen sulfide production. If it covers the butt of the tube, assume the butt is yellow

Citrate media

Citrate is the sole carbon source in this media. It is used to differentiate between the enterobacteria

Citrate pH indicator

bromothymol blue

Citrate results

A bright blue color indicates a positive result; the original media is blue-green and indicates a negative result

Carbohydrate fermentation tubes

This media contains a single carbohydrate per tube, plus a small, inverted test-tube called a Durham tube to catch any gas

Carbohydrate fermentation tube pH indicator

phenol red

Carbohydrate fermentation tube results

media must turn lemon-yellow to be positive, Durham tube must contain 25% gas to be positive.

MR-VP media

Methyl Red ,Voges-Proskauer tests for fermentation products, either acid or alcohol end products.

MR-VP pH indicator

methyl red; turns red at low pH

MR-VP results

MR positive indicates production of acid waste products. VP positive indicates production of an alcohol end product.

MR-VP reagents

After the bacteria has grown, methyl red pH indicator is added to the MR tube, and VP-A and VP-B are added to the VPtube.

SIM deep media

Used to differentiate enterobacteria according to motility, cysteine, and tryptophan digestion.

SIM inoculation

Use a sterile needle to pick up bacteria, stab straight down into media.

SIM results: motility

If the bacteria are flagellated, they will swim away from the stab line and produce cloudy media.

SIM results: S

If the bacteria can digest the cysteine, it will produce a black percipitate

SIM results: I

Production of Indole can be tested by adding Kovac's reagent to the top of the deep after incubation. A red ring is positive for indole production.

Urea slant

Contains urea

Urea slant pH indicator

Phenol red

Urea slant results

Bacteria that can digest the urea will grow, producing a fuschia color. If bacteria can not digest urea, the media stays the original color

Urea slant inoculation

Use a sterile loop to pick up a generous amount of bacteria. Inoculate with a fish-tail motion, bottom to top.

Phenylalanine slant

conatins the amino acid phenylalanine

Phenylalanine indicator

10 percent ferric chloride is added after incubation, a positive test will turn the slant a green color

Lysine or Ornithine broth

contain lysine or ornithine amino acids

Lysine or Ornithine broth pH indicator

bromocresol purple

Lysine or Ornithine inoculation

After steriley transferring bacteria, add 5 drops of oil to the top of the tubes.

Lysine or Ornithine results

A color change to purple is positve, yellow or the original color is negative