Quiz 3 Flashcards
What is Sticky test/KOH lysis test based on?
Based on the difference between the structure and thickness of the cell wall in Gram positive and Gram negative organisms and how it affects the ease of entry of chemicals into cells.
What does KOH do?
causes the lysis of cells by breaking down the cell wall and the cell membrane.
What happens if the cell wall gets thicker?
the thicker the cell wall, the harder it is for the KOH to penetrate
Purpose of Sticky test/KOH lysis test?
it will help you confirm whether or not your organism is Gram + or Gram -
What happens to Gram - under KOH test?
They lyse more quickly when treated with base and when they lyse, DNA is released and its viscous
What happens to Gram + under KOH test?
Gram positive cells take a lot longer to lyse because they have thicker cell walls which makes the KOH harder to penetrate.
What errors can occur in Sticky Test/KOH test?
If you leave the KOH on a slide with gram + too long or use old cultures of Gram + organisms, you are going to see lysis and this leads till a false interpretation of Gram -
What type of media is MacConkey?
it is both selective and differential
What is the purpose of using MacConkey medium?
its used to isolate and differentiate between members of the Enterobacteriaceae ( Gut Bacteria )
How is MacConkey like Levine EMB?
it allows a quick preliminary indication of whether a specimen contains enteric pathogens.
What does MacConkey include? and is it defined or undefined?
Its undefined ; and includes
- Lactose
- Protein
- Bile salts
- Neutral Red
- Crystal Violet
What 2 ingredients make MacConkey a selective media? and what do they select against?
- Bile salts
- Crystal violet
Both of them inhibit growth of Gram +
What 2 ingredients make MacConkey a differential Media?
- Neutral Red
2. Lactose
What is neutral red and what color to they exhibit and low and high pH
Neutral Red is a pH indicator.
- it is red at pH less than 6.8
- it is Colorless to yellow as pH increases
Explain the process of how neutral red works
A gram negative will ferment lactose -> this causes acid to build up -> The acid will decrease the pH –> the pH indicator will respond to this acid build up and change to a different shade of red depending on how much lactose is fermented
MacConkey results for COLONIES that are Gram Negative, Strong Lactose fermenter?
- these organisms produce a LOT of acid
2. Neutral red dye taken up by these organisms turn pink due to the decrease in pH, coloring the colonies RED/PINK
MacConkey results for AREA AROUND colonies that are Gram Negative, Strong lactose fermenters?
Acid that is excreted into the medium turns the neutral red dye in the medium around the colonies pink.
What happens when theres so much acid in the MacConkey medium?
this results to a large decrease in pH resulting in an opaque Bright pink halo due to precipitation of bile salts.
MacConkey results for COLONIES that are gram negative and regular lactose fermenters
Colonies are pink/red dude to decrease in pH
MacConkey results for COLONIES that are Gram Negative, NON-lactose fermenters?
Colonies are White or transparent that often turns into a brownish color.
Why are colonies a brownish color in the MacConkey results for a gram negative, non lactose fermenter?
it is due to a slight increase in pH. These organisms dont use lactose in the medium and uses the amino acids in peptone instead. Amino acid metabolism results in the release of ammonia, resulting in rise of pH
What is nutrient recycling?
The ability to break down large organic molecules such as nucleic acids, proteins, lipids, and polysaccharides that are released from dead plants/animal cells and reincorporate them into cellular material
What is required to break down large organic molecules?
It requires the production of extracellular enzymes
What are extracellular enzymes and what do they do?
They are enzymes that are made in the cells and released into the surrounding medium. These enzymes break down large molecules by HYDROLYZING the bonds and are known as hydrolytic enzymes
What is hydrolysis
Breaking bonds through the addition of a water molecule
Different types of Extracellular enzymes?
- Glycosidase
- Protease
- Esterase
- Nuclease
What does Glycosidase do?
Hydrolyze the glycosidic bond that binds monosaccharides
What are monosaccharides?
They are monomeric subunits of polysaccharides
What are exampls of polysaccharides
Celluloses, starches
What do proteases do?
hydrolyze peptide bonds that links amino acids
What do esterases do ? ( also known as lipases)
hydrolyze ester bond between glycerol and fatty acids in fats/oils
What do nucleases do?
Hydrolyze Phosphodiester bond between nucleotides of nucleic acids such as DNA and RNA
Even after Large organic molecules are broken down, can they still get into the cell membrane?
not necessarily. the cell membrane is a selectively permeable membrane that allows only certain molecules to enter.
How can the smaller components of nutrients enter the cell if it cannot go through the cell membrane?
The cell needs Special binding proteins on or in the membrane that recognizes and binds to these specific nutrients
What happens to the molecule once it binds to these special binding proteins? Whats this process called?
it is transported across the membrane into the cell, this is known as translocation
True or False, Translocation proteins are often specific to the particular molecule they are able to transport across the membrane
U is right…
Once the molecule is translocated, what conditions must it have to be able to contribute energy to the cell
- There has to be an appropriate metabolic pathway and
2. all the enzymes pertaining to the pathway are produced in the cell
What are carbohydrates?
Compounds containing Carbon, Hydrogen, and oxygen
Hydrated carbons