Micro Lab 2

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haleygreenbean
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309553
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Micro Lab 2
Updated:
2015-12-08 00:34:58
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microbiology
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micro 302 L
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  1. triglycerides or triacylglycerols
    • simple fats 
    • glycerol and three fatty acid chains
  2. phospholipid
    fat derivatives in which one fatty acid has been replaced by a phosphate group and one of several nitrogen-containing molecules.
  3. lipases
    enzymes that hydrolyze lipids
  4. triglycerides + H2O ---lipase---> glycerol + fatty acids
    lipolysis (lipid hydrolysis)
  5. rancidity
    rancidity testing determines the level of oxidation in a sample.
  6. SIM Agar
    • sim medium used to determine 3 bacterial activities:
    • 1. Sulfur reduction
    • 2. Indole production from tryptophan,
    • 3. Motility
  7. mixed acid fermentation
    • 1.anaerobic fermentation where the products are a complex mixture of acids, particularly lactate, acetate, succinate and formate as well as ethanol and equal amounts of H2 and CO2. It is characteristic for members of the Enterobacteriaceae family.
    • 2. MR test detects organisms capable of this
  8. gelatinase
    family of extracellular enzymes produced and secreted  by some microorganisms to hydrolyze gelatin
  9. H2S production
    turns grey , black , or blue
  10. gas production
    agar would be broken, cracked or pushed up in tube
  11. starch = amylose + amylopectin
    complex strand, branches off at multiple spots
  12. hydrolases
    break down large molecules in presence of water
  13. alpha -amylases
    break down big starch molecules
  14. metabolism
    • distinguishes bacteria 
    • sum of total reactions in bodies
  15. catabolism
    breaking down large molecules due to release in energy
  16. fermentation
    energy producing biochemical reactions usin electron donors from organic compounds
  17. indicators
    gentler indicator, phenol red, used for living cells, becomes bright yellow
  18. Durham tubes
    traps any gas produced, can tell whether or not its fermenting
  19. anabolism
    consume energy by building large molecules
  20. carbohydrate fermentation test
    • 1. put 3 different sugars into each tube with a bacteria
    • 2. Definition: hydrolysis of disaccharides prior to the fermentation reaction
    • 3. Goal: specific byproducts from different hydrolysis reactions indicates type of bacteria
  21. B-galactosidase test
    • 1. tests to see if B-galactosidase is produced by bacteria, which in turn indicates fermentation of lactose
    • 2. Definition: B-galactosidase is an enzyme that is produced to ferment lactose
    • 3. Test Result: 
    • a. use ONPG disc as lactose source, place in bacteria sample which goes in 37 C room.
    • b. will turn yellow due to a byproduct
  22. Triple Sugar Iron Test
    • 1. Purpose: to differentiate bacteria based on glucose, lactose or sucrose fermentation, or sulfur reduction
    • 2. whole tube will turn yellow if bacteria uses multiple sugars
    • 3. only yellow butt =glucose fermentation but not other sugars
    • 4. yellow = acidic 
    • 5. pink = alkalinic , or did not use a sugar but peptone and amino acids to alkalinize medium 
    • 6. H2S production: turns medium black, grey or blue
    • 7. gas production: agar would be broken, cracked, or pushed up in tube
  23. Starch hydrolysis test
    • 1. tests if bacteria has enzyme alpha amylase, which breaks down big starch molecules
    • 2. test itself:
    • a. put straight line of bacteria on a starch agar plate.
    • b. sat in 37 C room for at least 24 hours
    • c. after 24 hours, apply iodine onto starch agar.
    • d.positive for glucose production, of alpha amylase present is the "halo effect"
  24. oxidation/fermentation test
    • P.A.: w/ oil, green color; w/out oil, yellow color
    • a. means it is an obligate aerobe (needs O2)
    • E.coli: both w/ and w/out were yellow (doesn't need O2)
  25. litmus milk test
    • 1. Purpose: test for lactose fermentation, reduction of litmus, casein coagulation and casein hydrolysis
    • 2. Result: lavendar to white = lactose fermentation
  26. Lipid Hydrolysis Test
    • Purpose: to test for lipase productivity, an enzyme that hydrolyzes lipids
    • S. Aureus: colonies stay dark, halo of light indicates lipase activity (+ test)
    • Proteus mirabilis: colonies spread
  27. IMViC Tests
    • Indole
    • Methyl Red Test
    • Vogues-Proskauer Test
    • Citrate utilization (Simmons Citrate)
  28. Indole Test
    • Purpose: to see if organism can convert tryptophan, an amino acid, into indole
    • tryptophanase: an enzyme, breaks down tryptophan to make indole and pyruvic acid
    • Positive test: use indole reagent, pink ring = indole production
  29. Methyl Red test
    • Purpose: testing for mixed acid fermentation
    • Positive: pink/red color
  30. Voges-Proskauer test
    • Purpose: test for 2,3 butanadiol fermentation
    • since an alcohol, can't use an indicator
    • use methyl red and  acetoin instead (the product from fermentation)
    • Positive: pink red color due to acetoin production
  31. Citrate Utilization (simmons citrate)
    • Purpose: test to see if bacteria can use citrate as its only C source, end product is alkyl
    • Positive: if medium turns blue, positive for the products (pyruvic acid, CO2, and oxaloacetic acid)
    • ex/ that is positive: Salmonella typhimurium
    • tip: make sure lid is loose to allow CO2 to escape
  32. sulfur reduction, indole, and motility test (SIM test)
    • 1. Positive for motility and indole production: dark color
    • ex/ P.V. is motile
    • PM: grew everywhere like clouds
    • KO: single stabline w/ bugs
  33. Gelatin Hydrolysis test
    • Purpose: looking for bacteria that can produce gelatinase
    • Gelatinase: enzyme that breaks down gelatin
    • gelatin: protein fibers w/ matrix of water inside
    • Positive: if gelatin is liqueified, flowing freer than normal, proteus vulgaris is positive
    • TIP: will get FALSE + in 37 C
    • takes 10 days - 2 weeks
  34. Catalase test
    • Purpose: to test for catalase present
    • Positive: if H2O2 put on bacteria and bubbles, sign for the products water and free oxygen 
    • (means protection from red blood cells)
    • S. aureus is Positive!
  35. oxidase test
    • Purpose: identify bacteria w/ enzyme cytochrome c oxidase
    • cytochrome oxidase = found in electron transport chain
    • Positive: happens rapidly, will turn blue due to production of oxidized c cytochrome
  36. urease test
    • REAction: ammonia + phenol red + urease    --> ammonia, water and CO2
    • Purpose: to detect urease 
    • Positive: deep pink color from ammonia and phenol red
    • 24 hr test
  37. phenylalanine deaminase test
    • 1. tests bacteria's ability to make enzyme deaminase
    • 2. phenyl alanine + phenylalanine deaminase --> phenylpyruvic acid + ammonium ion + H2O
    • 3. phenylpyruvic acid + ferric Cl -> green complex
    • 4. Positive test: this green color NOT YELLOW-GREEN
  38. nitrate reduction test
    • Purpose: to test the ability that a bacteria can reduce nitrate (NO3) to nitrite (NO2) using the enzyme nitrate reductase. 
    • 2. if  red, positive for nitrate reduction
  39. agglutination reaction slide test
    • 1. put on square
    • 2. anitibody solution, 1 drop. 
    • 3. when antibodies are matched with their antigen 
    • 4. positive when beads form ( or clumping)
  40. double immunodiffusion (ouchterlony) test
    • (the too expensive test)
    • 2. put known antibodies around unknown antibody. 
    • 3. if match, makes division sign
  41. cocci
    round, spherical
  42. bacilli
    straight rod
  43. vibrio
    curved rods
  44. spirilla
    very curvy rods
  45. spirochetes
    long and slender bacteria
  46. square
    looks like thin crackers
  47. star
    found in water
  48. rosette
    found in water
  49. pleiomorphic
    variable, indistinct
  50. diplo
    pairs
  51. staphylo
    irregular clusters
  52. strepto
    in chains
  53. tetrad
    groups of 4
  54. sarcina
    3D
  55. palisade
    parallel
  56. gram negative stain
    thin walls, red or pink
  57. gram positive stain
    thick walls, blue or violet
  58. lag phase
    adjustment period, no cell division initially
  59. log phase
    growth rate takes off, exponential increase
  60. stationary
    reproduction rate is equal to the death rate
  61. death
    decline of organisms, more are dying than surviving- cannot do a viable plate count because you cannot tell which cells are living and which are dead
  62. generation time
    Time it takes the population to double
  63. mean growth rate constant
    The number of generations produced per hour
  64. experimental procedure
    • 1) Make your own blank- using distilled water- in a cuvetteKeep this refrigerated between uses
    • 2) Fill another cuvette ¾ full of the broth (this is your bug)
    • 3) Run the % trans and record result Keep this in the shaker bath between uses
    • 4) Do your serial dilutions and plate to find the countable CFU
    • 5) Repeat every hour, on the hour
  65. optimum growth rate
    the temp the bug will best grow at- classifications are made using this
  66. psychrophiles
    “cold lovers” live at 0-11 degrees C
  67. mesophiles
    “middle lovers” live at 12-44 degrees C
  68. thermophiles
    “heat lovers” live at 45 degrees and up
  69. extremophiles
    live way above, or way below the limits
  70. thermoduric
    don’t like high temps, but they can endure them
  71. acidophiles
    grow best at pH between 0-5.5
  72. Neutrophiles
    - 5.6-8.5
  73. Alkalophiles
    - 8.6-14
  74. buffers
    resist change in pH
  75. osmotic pressure
    pressure created when water goes from high to low concentration (usuallythrough a membrane)
  76. solvent
    dissolves
  77. water activity
    amount of water available for metabolism
  78. hypotonic
    there is more water outside of the cell than there is inside
  79. hypertonic
    more water inside of the cell than there is outside
  80. plasmolysis
    water moves from inside the cell to the outside of the cell so quickly it pulls awayand kills the cell
  81. isotonic
    having the same amount of water both inside and outside of cell – same osmotic pressure
  82. halophiles
    like high salt concentrations
  83. osmophiles
    like high sugar concentrations
  84. sporulation
    how bacteria survive extreme environments
  85. xerophiles
    love low water concentrations
  86. obligate aerobes
    require oxygen to live
  87. Facultative Anaerobes-
    grow better when oxygen is present but don’t need it to survive
  88. obligate aerobes
    presence of oxygen will kill them
  89. aerotolerant anaerobes
    don’t require oxygen to live- presence of oxygen doesn’t help or inhibit
  90. microaerophiles
    require a very small amount of oxygen to live
  91. anaerobic media
    media that lacks oxygen
  92. gas packs
    wisk away oxygen
  93. dimerization
    limitations of UV light
  94. light repair
    use energy from energy from visable light to repair what UV light killed
  95. dark repair
    use chemical reactions to repair damage from UV light
  96. disinfectants
    can be used on nonliving surfaces
  97. antiseptics
    gentle enough to be used on living surface
  98. microbicidal
    kills bugs
  99. micarobiostatic
    inhibits further growth but doesn’t kill the living
  100. phenol coefficient
    • relative effectiveness
    • <1 doesn’t kill as well as phenol
    • >1 kills better than phenol
  101. experimental procedure
    • 1) Add 0.1 ml bug to sterile saline
    • 2) Make a 0.1 ml spread plate
    • 3) Add 0.1 ml of selected disinfectant- This is the mix tube
    • 4) Add 0.1 ml to TBA at 0 min, 1 min, 5 min, and 10 min- after each TBA tube is made spread plate immediately!!
    • 5) Incubate at 37 degrees C
  102. kirby-Bauer method
    relative effectiveness of antibiotic- but doesn’t tell you how much youneed
  103. zones of inhibition
    if there is one the antibiotic is effective
  104. minimum inhibitory concentration
    what’s the lowest amount of the antibiotic that will do thejob- easy experiment- serial dilutions will tell you where to start your prescription

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