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BIO 275 > Practical 2 Exam > Flashcards

Practical 2 Exam Flashcards

1
Q

Blood Agar

A

Test for hemolytic microbes
a - green- partial hemolytic
b- clearing- complete hemolytic
y- no change- non hemolytic

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2
Q

Starch Agar

A

test for amylase
break down starch into maltose
clearing - positive for presence of amylase

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3
Q

Casein Agar

A

test for presence of casein (milk protein)
break protein into small peptide
clearing means microbe produce casein to break down protein

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4
Q

Spirit Agar test

A

presence of lipase
break lipid into fatty acid
clearing means microbes able to produce lipase to break down lipids

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5
Q

DNAses

A

presence of DNAses
break DNA into small fragment
clearing means microbes able to break down DNA

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6
Q

MacConkey Agar

A
  1. selective and differential media
  2. prevent gram + organisms from growing
  3. allows differential between microbes based on their biological characteristics
  4. use neutral red as pH indicator
  5. red means gram - microbes and able to ferment lactose while no change means non fermenters.
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7
Q

Mannitol-Salt Agar

A
  1. selective and Differential media
  2. allows halophilic and halotolerant organisms to grow
  3. use phenol red as a pH indicator
  4. yellow means fermenters of mannitol/sugar while white means growth/halotolerant but not fermenters.
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8
Q

Hektoen Enteric Agar

A
  1. use bromothymol blue and acid fuchsin as a pH indicator
  2. presence of fructose, salicin, and sucrose
  3. determine which metabolic pathway is used to ferment sugar
  4. green is a non-fermenter while red/orange/yellow fermenters
  5. thiosulfate and ferric ammonium citrate (H2S producers are black)
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9
Q

IMViC test

A

determine the type of organism based on biochemical reaction

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10
Q

Indole Test

A

uses indole reagent
a red ring at the top of the media means positive for Indole
used to determine if the microbe is able to produce tryptophanase and break down tryptophan into indole

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11
Q

Methyl Red Test

A

identify mixed acid glucose fermenters
determine which metabolic pathway is used by microorganisms to ferment sugar
methyl red is added and if the color change (red) means the acid produced is stable

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12
Q

voges proskauer test

A

able to ferment glucose
test for the presence of acetoin produced from pyruvate in the butanediol pathway.
red means positive for acetoin after a-naphthol and KOH creatine is added.

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13
Q

Simmon Citrate Agar/test

A

use to determine if citrate is the sole carbon source for microbe
blue means positive
green means negative.

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14
Q

EnteroPluri Test

A

series of 15 compartmentalized test that determines if there is contamination present
identify gram (-) oxidase (-) rod organism

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15
Q

TSA

A

control used for oxidase/catalase test

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16
Q

catalase test

A

determines if catalase is present
when hydrogen peroxide is added and bubbles formed means positive for catalaze enzyme
host produce catalase to protect from ROS, catalase break down hydrogen peroxide into water and oxygen.

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17
Q

oxidase test

A

identify if cytochrome c is present
turns dark purple/blue when cytochrome c is reduced after adding oxidase reagent

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18
Q

TDT

A

thermal death time
the shortest period of time need to kill all microbes at standard condition

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19
Q

TDP

A

thermal death point
the lowest amount of temp. in which all microbes killed after 10-minute exposure.

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20
Q

incineration

A

control microbe growth through dry or moist heat, boiling water and pasteurization

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21
Q

dry heat

A

oxidizes cell components

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22
Q

moist heat

A

denature protein and nucleic acid

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23
Q

autoclave

A

use high heat and moisture

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24
Q

endospore

A

can resist high temp.

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25
Q

UV light

A
  1. has sterilizing ability but has limited penetrating power
  2. cause thymine dimers if not repaired can cause cell death
    DNA polymerase and ligase repair UV damage cells via dark repair. Polymerase synthesizes new DNA while ligase joins new and old DNA in photoreactivation.
  3. pigmented cells and endospores resist UV exposure.
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26
Q

osmotic pressure

A

when two solutions of different solute concentrations are separated by a semipermeable membrane

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27
Q

movement of water

A

in and out of the cell can affect cell growth/survival

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28
Q

osmophilic organisms

A

can grow in high osmotic concentration

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29
Q

what metals have toxic effect on microbes

A

copper, mercury, and silver

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30
Q

silver nitrate

A

used in clinical setting to reduce bacterial growth
used on baby eyes as antiseptic

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31
Q

increase in temperature

A

as temperature increase, non-endospores has no growth after exposure to high heat.
the temperature at 100 degrees was for 10 minutes while 37, 63, and 80 exposed at 30 minutes)

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32
Q

UV light and microbe growth

A

endospore was able to growth after exposure to endospore. others were not due to thymine dimers and inability to repair DNA.

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33
Q

osmotic concentration

A

as salt concentration increase organisms slowly die leaving only those that are halophilic/halotolerant to still grow.

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34
Q

heavy metal

A

clearing around metals to prevent growth

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35
Q

DNA ligase

A

enzymes that joins short pieces of DNA

36
Q

DNA polymerase

A

enzymes that joins nucleotide of DNA

37
Q

endonuclease

A

enzyme that attack the internal region of nucleic acid (covalent bond between nucleotide)

38
Q

mutagen

A

any chemical or physical agent that increase the mutation rate

39
Q

mutation

A

a permanent genetic change

40
Q

purine

A

nitrogen containing ring base found in nucleic acid (adanin and guanine)

41
Q

pyrimidine

A

a nitrogen containing ring base found in nucleic acid (cytosine, thymine, and uracil)

42
Q

thymine dimer

A

a compound resulting from the joining of two adjacent thymine molecules. cause distortion in DNA molecules.

43
Q

inhibition zone

A

clearing (oligodynamic) around metal or antibiotic where there is not growth

44
Q

disinfectant

A

chemical that kill microbes and are applied to inanimate objects

45
Q

antiseptic

A

chemical that kills or inhibit microbes and can be applied to living tissues (non-toxic)

46
Q

antibiotic

A

group of biproduct of microbes that are able to kill other microbes

47
Q

MIC

A
  1. minimal inhibitory concentration
  2. the smallest amount of antibiotic needed to inhibit the growth of a microorganism
  3. recognized by clear inhibition zones around the antibiotic disk
48
Q

how do antibiotic inhibit

A

cell wall synthesis, protein synthesis, metabolism, or nucleic acid synthesis

49
Q

PCN

A
  1. has beta lactam ring structure
  2. prevent cell wall division (on growing bacteria)
50
Q

tetracycline

A
  1. has a ring structure
  2. bind 30S ribosomal subunit
  3. interfere with protein synthesis
51
Q

sulfonamides

A
  1. sulfa drugs look like another molecules (p-aminobenzoic acid precursor to folic acid)
  2. unable to make folic acid, no amino acid made, then nucleotide can’t be made.
52
Q

quinolone

A
  1. has 4 quinolone ring
  2. block nucleic acid synthesis by inhibiting gyrase and topoisomerase II
53
Q

obligate intracellular parasite

A

alters host cells they infect (must be inside bacteria cell)

54
Q

binal shape

A

has polyhedral head and helical tail

55
Q

T (type) coliphages

A

most-studied bacteriophage that affects E. coli B

56
Q

bacterial phage multiplication

A
  1. depends on viral structure and genome
  2. a. attachment to host cell
    b. nucleic acid entry
    c. synthesis
    d. assembly
    e. release
57
Q

lytic acitivity

A

can be observed by clear zone (plaque)

58
Q

plaque assay

A

use dilution of prepared virus plated on lawn of host cells

59
Q

plaque forming units

A

(PFU)
number of plaque counted and results expressed

60
Q

memory

A

ability to respond more vigorously when reexposed to the same antigen

61
Q

tolerance

A

ability to discriminate self antigens from nonself antigen

62
Q

immunogen

A
  1. foreign macromolecules that induce an immune response
  2. when introduced into a host in an appropriate dose and route, they initiate an immune response (vaccine).
63
Q

intrinsic property of immunogens

A

molecular size (larger), complexity (linear or globular), and physical form (folded, certain cavity)

64
Q

conformational determinant

A

a segment/sequence on the antigen that antibodies are able to recognize

65
Q

antigenic determinant (epitope)

A

a distinct portion of macromolecule on the antigen recognized by the immune system

66
Q

antibody protein structure

A

has two heavy chain and two light chain

67
Q

amino-terminal region

A

variable domain (differs in each different antibodies)

68
Q

antigen binding site

A

formed by interaction of variable regions of heavy and light chains
come together to form pocket and allow binding of anitgen

69
Q

anitbody production

A
  1. inject goat with a primary injection containing antigen (bovine albumin (this produce small antibody)
  2. inject booster after 2 weeks (increase antibodies, longer response time)
  3. collect blood after 2 weeks
  4. clot and centrifuge blood, discard cells, and isolate antiserum.
70
Q

Naming antibody

A

GOAT ANTI-BOVINE ALBUMIN ANTIBODY
1. animal antibody raised in
2. antibody against/bind to
animal injected GOAT
against (anti)
what was injected BOVINE ALBUMIN
what is produced ANTIBODY

71
Q

Lettice hypothesis

A
  1. the right antigen-to-antibody ratio forms a lattice that is insoluble forming a precipitate
  2. precipitate forms a white line (proper concentration)
  3. no precipitate means no antigen and antibody reaction
72
Q

antibodies are BIVALENT

A

combine with two antigen

73
Q

antigens are POLYVALENT

A

combine with many antibodies (5-6)

74
Q

bacterial transformation

A

process by which bacterial cells take up naked DNA molecules
first done by Griffith

75
Q

transformation effect

A

organism able to pick up new traits that improve the ability to survive.

76
Q

plasmid

A
  1. DNA outside chromosome
    2.encode protein for antibiotic resistance, as well as metabolic pathway.
77
Q

insulin

A

no longer extracted from animal pancreas but expressed in bacteria

78
Q

transformation step

A
  1. preincubation step: cell suspended in cation solution and at low temp.
    a. reduce (-) charge of the membrane (neutralize)
    b. loosen membrane making it competent
  2. Add DNA (plasmid)
    a. competent cell allows the plasmid to move closer to the cell.
  3. Heat shock cell (42 degrees Celsius)
    a. heating creates draft
    b. pulls DNA into the cell (maximize movement)
  4. Recovery Step
    a. membrane becomes less loose trapping plasmid inside the cell
    b. incubated with LB broth and plated into media.
    c. transform cell begins to divide and replicate new plasmid
79
Q

LB broth and LB agar

A

with and without ampicillin
ampicillin inhibits bacterial growth

80
Q

pGreen plasmid

A

contains a gene that codes for green fluorescent protein (GFP)
a gene encoding beta-lactamase, an enzyme that breaks down ampicillin

81
Q

bacterial conjugation

A
  1. process by which bacterial cells transfer DNA molecules through direct contact or sex pili
  2. sex pili is hallow and DNA travel from donor to recipient
  3. pilus work as lasso and pulls recipient close (F-pilus get unidirectional DNA transfer from donor to recipient)
82
Q

E. coli Strain I

A

Streptomycin resistant

83
Q

E. coli Strain II

A

ampicillin resistant

84
Q

general transduction (E. coli Strain B E)

A
  1. E. coli exposed to phage with ampicillin-resistant gene
  2. plate new strain of bacteria on ampicillin plate
  3. any part of the bacterial genome can be transferred (lytic cycle)
  4. during viral assembly, fragments of DNA mistakenly get assembly into the phage head
  5. transducing phage injects transduced DNA into new bacteria.
85
Q

specialized transduction

A
  1. carried out by temperate phage that has established lysogeny
  2. only a portion of the bacterial genome is transferred.
  3. occurs when prophage is incorrectly excised.
86
Q

transformation experiment

A
  1. calcium chloride used
  2. E. coli added and placed back in the ice
  3. DNA plasmid added to the tube and placed in ice for 15 min
  4. heat shock cell at 42 degrees Celsius for 90 sec.
  5. tube return to ice for 1 minute.
  6. LB broth was added to the tube and left at room temp for a 10-minute recovery.
  7. broth is placed in an LB amp plate using a spreader.
  8. let plate rest until suspension become absorbed into agar.
  9. placed in incubator for 24-36 hours.
87
Q

conjugation experiment

A
  1. E coli strain I and II place on LB plate both grew
  2. placed on LB amp plate. only E coli II grew
  3. placed on LB Str plate. Only E coli I grew
  4. Both placed on LB mating plate using spreader. when place in AMP/STR plate was resistant to both

BIO 275 (9 decks)

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