Farmingdale State College
Professor Bissoon
Medical Biology
Bio 318L
Spring 2021
Enteric assignment
Answer the follow questions (worth 20 points)
1. What is the purpose of the MacConkey Media? What is a positive and negative results? List
some gram-negative bacteria that will grow on this medium.
MacConkey Agar (MAC) is a selective and differential medium used to separate and distinguish enteric
bacteria based on their lactose fermentation capacity. Gram-positive bacteria are inhibited by bile salts
and crystal violet. Lactose is a fermentable carbohydrate source that allows for distinction.
Lactose fermenting organisms will look pink as the neutral red becomes red, resulting in an acidic
environment. Because of the pH transition, bile salts can precipitate out of the media surrounding the
growth of fermenters. Non-fermenters will develop colonies that are usually colored or colorless.
Salmonella, Proteus colonies, Yersinia, Pseudomonas aeruginosa, and Shigella are examples of non-
lactose fermenting bacteria.
2. What is the Hektoen Enteric media? What is in the media that makes it selective? What is a
positive and negative result?
Hektoen Enteric Agar (HE) is a selective and differential medium used to separate and distinguish
Salmonella and Shigella bacteria from other Enterobacteriaceae bacteria. Many Gram-positive species
are inhibited by bile salts, as well as the dyes bromothymol blue and acid fuchsin.
Lactose, sucrose, and silicon are fermentable carbohydrates that help enteric bacteria develop and
differentiate. Sulfur is obtained from sodium thiosulfate. Ferric ammonium citrate serves as an iron
source, allowing hydrogen sulfide to be generated from sodium thiosulfate, which serves as a sulfur
source. By reacting with hydrogen sulfide gas to form a black precipitate, ferric ammonium citrate also
enables the visualization of hydrogen sulfide development.
Enterics that ferment one or more of the carbohydrates create colonies that range from yellow to
salmon in color. The colonies of non-fermenters would be blue/green. Black colonies or blue/green
colonies with a black core are produced by organisms that convert sulfur to hydrogen sulfide.
Farmingdale State College
Professor Bissoon
Medical Biology
Bio 318L
Spring 2021
3. What is the EMB media? What is a positive and negative result? What color does E.coli
produce?
Eosin methylene blue agar is a selective and differential medium used to isolate fecal coliforms. At low
pH, the pH indicator dyes eosin Y and methylene blue combine to create a dark purple precipitate,
which inhibits the growth of most Gram positive bacteria. Sucrose and lactose are fermentable
carbohydrate sources that promote the growth of fecal coliforms by still allowing them to be
distinguished.
Lactose or sucrose fermenters that are active can contain enough acid to create the dark purple dye
complex. These species' development would be dark purple to black in color. Mucoid pink colonies are
produced by slow or poor fermenters. Colonies that are normally colored or colorless suggest that the
organism should not ferment lactose or sucrose and is not a fecal coliform.
The color E. coli would produce is a green metallic sheen.
4. What is TSI? What are the different results that can form?
The Triple Sugar Iron (TSI) test is a microbiological assay that evaluates a microorganism's ability to
ferment sugars and generate hydrogen sulfide. It is sometimes used to distinguish enteric bacteria such
as Salmonella and Shigella.
Different results that can form the TSI test are:
Red/yellow- Glucose fermentation only, peptone catabolized.
Yellow/yellow- Glucose and lactose and sucrose fermentation.
Red/Red- No fermentation, peptone catabolized.
Red/ no color change- No fermentation, peptone used aerobically.
Yellow/yellow with bubbles- Glucose and lactose and sucrose fermentation, Gas produced.
Red/ yellow bubbles- Glucose fermentation only, Gas produced.
Red/ yellow with bubbles and black precipitate- Glucose fermentation only, H2S produced.
Yellow/ yellow with black precipitate- Glucose and lactose and/or sucrose fermentation, H2S produced.
No change/no change- No fermentation
5. What is an API strip? and Why is it used?
Farmingdale State College
Professor Bissoon
Medical Biology
Bio 318L
Spring 2021
The API strip is a set of biochemical tests, such as enzyme and fermentation tests. For the biochemical
experiments, each tube comprises dehydrated substrates. The findings are dependent on color
variations within each tube following the incubation period. A tube and a cupule make up each
microtube. This procedure is used to determine the presence of enteric gram-negative rods. On the
strip, there are 20 different evaluation compartments.
