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Microbiology 1 Flashcards

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1
Q

Microbiology

A

the study of microscopic forms of life

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2
Q

Microscopic groups

A

bacteria, fungi, protazoa, viruses

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3
Q

Pathogenicity

A

certain mircobes cause disease, learning how diseases are transmitted is important to health sciences.

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4
Q

Bioremediation

A

process using naturally occurring or genetically engineered microorganisms to degrade or detoxify environmental hazards (ex. oil spills)

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5
Q

Microbes biological roles

A

are essential to web of life, some are photosynthetic, others decompose dead organisms and some provide nitrogen to plants

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6
Q

Fundamental biology

A

microbiology provides insight into life processes of all life forms

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7
Q

Microbes

A

are useful in research, are prokaryotes

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8
Q

antibiotics

A

can be isolated from microbes (ex penicillium notatum)

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9
Q

genetic engineering

A

microbes can be used to create useful human proteins (ex insulin and HGH)

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10
Q

Characteristics of microbes

A

ubiquitous, microscopic, unicellular (some are multicellular), sexual and/or asexual reproduction, growth in number, metabolic activity, free living (some are parasitic), respond to stimuli, mutations, range in size from 20nm to 100um

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11
Q

Domains

A

Domain Archaea (extremeophiles), Domain Bacteria, Domain Eukarya

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12
Q

Microbe types

A

Bacteria, Algae, Fungi, Viruses, Protazoa

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13
Q

Bacteria

A

are unicellular, microscopic, some are photosynthetic, no nucleus, have 3 types of morphology

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14
Q

Bacteria shapes

A

round, rod, tight spiral, loose spiral, comma shaped

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15
Q

Round bacteria

A

cuccus, cocci

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16
Q

Rod shapes bacteria

A

bacillus, bacilli

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17
Q

Tight spiral shaped bacteria

A

spirochetes

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18
Q

Loose spiral shaped bacteria

A

spirillum

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19
Q

Comma shaped bacteria

A

vibrio

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20
Q

Algae

A

unicellular or multi-cellular, eukaryotic

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21
Q

Fungi

A

kingdom fungi, in domain eukaria, unicellular yeasts, multi-cellular molds, macroscopic (mushrooms), are an important source of antibiotics

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22
Q

Mushrooms

A

absorb their nutrients and decompose, no photosynthesis

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23
Q

Viruses

A

not living things, need host cell to reproduce, are more like complex chemicals, have neuclaic acid genome and protein capsid-“neucleoproteins”, obligate intracellular parasites

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24
Q

Protazoa

A

Eukaryotic organism, has many vacules and organelles, are motile, ameba

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25
Q

Robert Hooke & Anton Van Leeuwenhoek

A

Invented the first microscope, first to see living organisms under microscope, 1600’s. Hooke coined term “cell”

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26
Q

Carolus Linneaus

A

developed binomial nomenclature by using genus and species. Genus is capitalized, species is not, 1735

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27
Q

Compound light microscope

A

system of magnifying lenses arranged to produce an enlarged image of an otherwise invisible object, has two lenses (ocular and and objective)

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28
Q

Theodore Schwann

A

developed the cell theory

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29
Q

Cell theory

A

cells are the fundamental units of life and carry out the functions of living things

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30
Q

Germ theory of disease

A

microorganisms can invade other organisms and cause disease

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31
Q

Spontaneous generation

A

the belief that life can spontaneously arise from non-living matter

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32
Q

Redi

A

1668, disproved the theory that maggots come from meat

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33
Q

Needham

A

boiled chicken broth and put it into different containers to prove spontaneous generation for microbes in 1745

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34
Q

Spallanzani

A

repeated Needham’s experiment but containers, no bacteria formed, critics said it was because of lack of oxygen

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35
Q

Pasteur

A

1861, disproves spontaneous generation and proves biogenesis, also developed aseptic technique, pasteurization, rabies vaccine, fermentation

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36
Q

Biogenesis

A

all life comes from living things

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37
Q

Robert Koch

A

1884, developed pure culture technique

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38
Q

Koch’s postulates

A
  1. the specific causative agent must be found in every case of disease
  2. the disease organism must be isolated in pure culture
  3. inoculation of the culture into healthy susceptible animal must produce the same disease
  4. the disease organism must be recovered from the inoculated animal
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39
Q

Semmelweiss

A

1840’s, hand washing, pioneer of aseptic technique

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40
Q

Mortality and handwashing

A

handwashing use lowered mortality rate fom 10-35% to 1% in maternity ward

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41
Q

Lister

A

1860’s developed use of chemical disinfectants in surgery

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42
Q

Alexander Flemming

A

1928, discovered penecillin

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43
Q

Chemotherapy

A

treatment of a disease with a chemical

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44
Q

Antibiotics

A

microbe made chemicals

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45
Q

Synthetics

A

man made chemicals

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46
Q

Microscpoe

A

a system of magnifying lenses arranged in an order to provide a visible image of an otherwise non-visible object

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47
Q

Polio Virus size

A

20nm (0.02um)

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48
Q

Small Pox Virus size

A

300nm (0.3um)

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49
Q

Staphylococcus aureus

A

0.5um, typical coccus shape (round)

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50
Q

Pseudomonas aeruginosa

A

5um long, typical bacillus shape (oblong)

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51
Q

Treponema pallidum

A

20um long, very thin, spirochete shape

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52
Q

Resolution

A

R= λ / 2 x NA

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53
Q

NA

A

numerical aperture, a mathematical expression relating to the extent that light is concentrated by the condenser lens and collected by the objective

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54
Q

λ

A

wavelength, is directly proportional to resoulution, as the wavelength decreases, the resolving power increases

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55
Q

Important features of a microscope

A

magnification, resolution

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56
Q

Resolution

A

smaller numbers make a clearer image, is a number that indicates the smallest microbe or part of a microbe that can be viewed with clarity

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57
Q

Light microscopy

A

uses light as the illuminating source, uses glass lenses, λ=550nm

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58
Q

Electron microscopy

A

uses electrons as illuminating source, uses electromagnetic lenses, λ= 0.005nm, has 100,000 fold resolution than light microscopy

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59
Q

Light properties

A

light can be reflected, transmitted and absorbed, we want to avoid refraction

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60
Q

Index of refraction

A

speed at which light passes through material

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61
Q

Microscope types

A

compound bright field (compound light microscope), dark field, phase contrast, fluorescence, electron

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62
Q

Compound bright field

A

field of view is brightly lit, magnification=2,000x, resolution with oil objective=0.2um

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63
Q

Compound bright field uses

A

clinical, research, fixed and stained microbes

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64
Q

Compound bright field limitations

A

poor image production of viable microbes, can not see anything under resolution of 0.2um

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65
Q

Dark field microscope

A

light reflected off surface of specimen to view, background is dark, magnification=2,000x, resolution with oil objective=0.2um

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66
Q

Dark field uses

A

image production of viable microbes that are not easily stained or cultured in vitro
Major use: diagnosis of tetranemapalidum (syphilis) which is a clinical use

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67
Q

Dark field limitations

A

poor image production of classically fixed and stained microbes, can not see anything under resolution of 0.2um

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68
Q

Phase contrast microscope

A

dual beam-light comes from two sources
magnification=2,000x
resolution with oil objective=0.2um

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69
Q

Phase contrast uses

A

highly contrasted images of internal structures of visible microbes including flagella and cilia

70
Q

Phase contrast dual beam

A

can cause reinforcement or interference because light comes directly from light source and is also refracted off the specemin

71
Q

Phase contrast limitations

A

poor image production of classically fixed and stained slides, can not see anything under resolution of 0.2um

72
Q

Fluorescence microscope

A

AKA ultraviolet or immunofluorescent, uses ultraviolet light, magnification=2,000x
Wavelength of UV light is between 100-400um making for better resolution

73
Q

Fluorescence microscope uses

A

look at microbes with natural fluorescence or those that selectively bind to flurochains

74
Q

Pseudomonas aeruginosa

A

is naturally fluorescent, best used with fluorescence microscope

75
Q

Mycobacterium tuberculosis

A

binds to auromine fluoricome and appears bright yellow under fluorescence microscope

76
Q

Bacillus enthracs

A

binds tofluorescin isothioyanate (FITC) to make apple green color under fluorescence microscope

77
Q

Acridine orange

A

binds to neulcaic acids like DNA

78
Q

Fluorescence microscope major use

A

immunofluorescence

79
Q

Fluorescence microscope clinical use

A

fluorescent antibody, used to diagnose rabies and syphylus

80
Q

Fluorescence microscope research

A

need to make an antibody and combine it with fluorocrome, can use fluorescent antibody to bind to a sample

81
Q

Electron microscopy

A

Transmission (TEM) and Scanning (SEM)

82
Q

Transmission microscopy

A

transmission electron microscope, magnification=100,000x, resolution=2.5nm (0.0025um)

83
Q

Transmission microscopy uses

A

research for fixed and stained microbes, no size limitation

84
Q

Scanning microscopy

A

produces 3D image, magnification=10,000x, resolution=10nm

displaces electrons from the surface of cells, secondary electrons are responsible for creating shape

85
Q

TEM slide prep

A

takes days to prep slides which keeps use limited

  1. grow culture of bacteria overnight
  2. fix bacteria in “aldahide” substance
  3. sequential dehydration using alcohol
  4. embed cells into a resin
  5. thin sections of resin using microtome (sections are 10-50nm)
  6. float sections onto copper mesh where sections are stained using heavy metal
86
Q

Transmission microscopy limitations

A

preparation can cause distortion of the sample causing mesosomes which are artifacts

87
Q

Microscope slide techniques

A

Wet mount
smears
staining
hanging drop-is good for viewing motility

88
Q

Simple stain

A

used to see morphology and arrangement

ex: crystal violet

89
Q

differential stain

A

can differentiate organisms and place microbes into major taxanomic groups

90
Q

gram stain

A

type of differential stain, can see composition of cell wall, can be gram positive or gram negative

91
Q

acid fast stain

A

trying to detect certain cell wall component (myloric acid)

Only myobacterium and nocardia are acid fast bacteria

92
Q

Special stain

A

try to detect presence and/or location of certain structure

93
Q

acidic dye

A

is a special stain, dye molecule is associated with the negatively charged ion. It is a negative stain.
Won’t stain bacteria because they already have a net negative charge

94
Q

Negative stain

A

is a special stain, will stain the background and then you have to go back and stain the bacteria

95
Q

Flagellar stain

A

is a special stain, can see flagella, very difficult stain to achieve, can see presence and location of flagella

96
Q

Endospore stain

A

is a special stain, used to identify the presence of endospores

97
Q

Taxonomy

A

the classification of bacteria into major groupings based on observable, measurable traits
Similarities are presumably due to relatedness

98
Q

systemats

A

the study of the evolutionary relationship of organisms, can be proven by DNA sequencing

99
Q

5 kingdom classification

A

plantae, fungi, animalia, protista, monera

100
Q

Domain eukaria

A

contains, plantae, fungi and animalia

101
Q

Kingdom protista

A

no longer recognized, is being broken down into additional kingdoms using rRNA sequencing
organisms in this kingdom are very nutritionally diverse, contains all unicellular animals and unicellular plants

102
Q

Kingdom monera

A

no longer recognized, contained all bacteria and archea, is divided into domain archea and domain bacteria

103
Q

Prokaryotic species

A

population of cells with similar characteristics because they are all descendants of a single parental cell
clonal population=pure culture

104
Q

Bacterial strain

A

ex: E. Coli 0157H7
Numbers and letters indicate strain type
Strains are created by picking up extra genes

105
Q

Growth media function types

A

Liquid (broths)
solid (agar plates, slants, deeps)
semi-solid (tubes)

106
Q

General purpose media

A

contains nutrients that allow certain types of bacteria to grow
ex: TSA (trypticase soy agar)

107
Q

Selective media

A

contains a selective agent that only allows certain types of bacteria to grow
ex: MSA, Sabdex

108
Q

MSA

A

mannitol salt agar, contains a high concentration of salt, is selective for certain organisms

109
Q

Sabdex

A

high dextrose concentration, low pH which favors the growth of fungi over bacteria making it selective but it is also a general purpose media for growing fungi

110
Q

Enriched media

A

contains specific growth factors and/or vitamins that needed by bacteria that can’t grow on general purpose media
ex: blood agar

111
Q

Differential media

A

contains a constituent that causes an observable change (in color or pH) in the medium when a particular chemical reaction occurs
this makes it possible to distinguish between organisms

112
Q

MSA

A

type of growth media, contains mannitol, salt and phenol red (pH) indicator
pH>6.8=red
pH<6.8=yellow

113
Q

Magnification

A

increased size of a virtual image

114
Q

Resolution

A

increased clarity or definition of detail or virtual image

115
Q

Light microscopy

A

refers to microscopy that uses visible light to observe specemins

116
Q

Image production pathway

A

illuminator
condenser (which directs light through the specimen)
objective lens(which magnifies the specimen)
occult lens (magnifies the specimen again x10)

117
Q

Total magnification

A

Objective lens X ocular lens

118
Q

Working distance

A

distance between object on the slide and the objective

119
Q

Refractive index

A

refers to the light bending properties of differential substances

120
Q

Oil immersion

A

used to reduce refraction, has same refractive index of glass, acts as an extension of the objective lens

121
Q

Smear slide

A

before we stain any slide we need to prepare the smear

122
Q

Smear slide objective

A

bind cells to the slide
inactive cells
facilitate stain penetration

123
Q

Measuring microbes

A

at 100x distance between 2 lines is 1um

at 40x distance between 2 lines is 2.5um

124
Q

Scientific method

A 
observation/research
hypothesis
prediction
experimentation
results
conclusion
125
Q

Observations

A

things we see in nature
hear/read about
possibly research the topic

126
Q

Hypothesis

A

provides a solution to our observations
an”educated guess”
a good hypothesis is testable

127
Q

prediction

A

hypothesis can take the form of prediction
uses and if-then statement
allows one to specify how the hypothesis will be tested

128
Q

experiment

A

testing your hypothesis

129
Q

Independent variable

A

intentionally changed

130
Q

dependent variable

A

the value we measure from the experiment

131
Q

Controlled variables

A

are held constant

132
Q

Control treatment

A

should vary from the experimental group by only one variable or it will be hard to determine what caused results
the independent variable is eliminated or set to a standard value

133
Q

Controls

A

positive control

negative control

134
Q

Positive control

A

an experiment whose results are known to be positive

135
Q

Negative control

A

an experiment whose result is known to be negative

136
Q

Results

A

data should be presented in tables and graphs for analysis

data should be able to “stand alone”

137
Q

conclusions

A

based on results, either supports or does not support the hypothesis
you don’t “prove” anything from just one experiment

138
Q

Mushrooms

A

are not plants, they do not preform photosynthesis

139
Q

Kingdom plantae

A

multi-cellular photosynthetic autotrophes (self nourishing)

140
Q

Kingdom anamelia

A

heterotrophic consumers

141
Q

Domain bacteria

A

all pathogens, also non pathogenic bacteria from soil and water

142
Q

Domain archea VS domain bacteria

A

domain archea do not have peptidoglycan in their cell walls and domain bacteria do

143
Q

Domain archea

A

Have 3 groups
Methanogens
Halophiles
Hyperthermophiles

144
Q

Methanogens

A

make methane
are strict anerobes(need anerobic environment
CO2 + H2 –> OH4 (methane)

145
Q

Halophiles

A

require high salt consentration

obligate/strict halophiles

146
Q

Hyperthermophiles

A

grow in extremly hot environments

proteins have adapted and do not denature in heat

147
Q

Taxonomic study steps

A

establish a pure culture
characterization of a pure culture
DNA hybridization

148
Q

How to establish a pure culture

A

must use streaking, “ streak plate method”

EX: onto an agar plate

149
Q

Characterization of a pure culture

A 
microscopic characteristics
cultural characteristics
biochemical characteristics
chemical characteristics
genetic characteristics
150
Q

Microscopic characteristics of a pure culture

A

use a gram stain to see (AKA differential stain)

151
Q

Cultural characteristics of a pure culture

A

environmental factors like temperature, O2 required and pH required
colony characteristics like pigment, opacity, margins and elevation

152
Q

Biochemical characteristics of a pure culture

A

best way to determine unknown

uses dichotomous key (we will make our own)

153
Q

Chemical characteristics of a pure culture

A

immunological, use ELIZA test

154
Q

ELIZA test

A

enzyme linked immunosorbent assay

used to determine chemical characteristics of microbes

155
Q

Genetic characteristics of a pure culture

A

use ribosomal gene sequencing

156
Q

Ribotyping ribosomes

A

composed of protein and ribosomal RNA, highly conservative (very few evolutionary change over time), all members of a certain domain, may have signature sequences

157
Q

Facultative anerobe

A

can grow in oxygenated or non-oxygentaed environment unlike a strict anerobe

158
Q

Klebsiella

A

large mucoid colonies

159
Q

Genus protius

A

has swarming motility

160
Q

Phenol red

A

if pH is > 6.8, it is a red color

if bacteria ferments lactose to acid products, pH will drop and color is yellow

161
Q

DNA hybridization

A

heat DNA to break hydrogen bonds between nucleotides

only used to exclude relatedness, can’t say things are related based on their G+C ratio

162
Q

DNA composition

A

base composition is a fixed property
G base pairs with C, A base pairs withT
% of G+C gives you, in theory, A’s and T’s

163
Q

DNA hybridization method

A
  1. purify DNA from unknown
  2. heat double strand of DNA until it is a single strand
  3. monitor process using spectrophotometer
  4. determine melting temperature
  5. based on melting temp, we get % of G+C, compare % to other known organisms
164
Q

DNA hydrogen bonds

A

A’s and T’s have 2 hydrogen bonds and melt at a lower temperature
G’s and C’s have 3 hydrogen bonds and melt at a higher temperature

165
Q

Gram positive cell

A

has thick peptidoglycan layer
physically stronger
has small “pores”

166
Q

Gram negative cell

A 
has thin peptidoglycan layer surrounded by outer membrane
chemically stronger (outer membrane resists penetration of chemicals)
large "pores"
167
Q

Gram positive cell stain

A

purple

CV-I complex is larger than pores, alcohol dehydrates and further shrinks peptidoglycan, trapping purple stain

168
Q

Gram negative cell stain

A

pink
alcohol dissolves the outer membrane and CV-I complexes wash out of larger pores
counter-stain pink with safranin

169
Q

Gram type

A

differences in cell’s wall

170
Q

Gram reaction

A

the color it shows when stained

171
Q

Gram-Variable

A

gram type and gram reaction do not always correlate

technique and age of the culture can affect the reaction

172
Q

KOH test

A

can be used to confirm gram stain test
gram-negative bacteria will lyse in the alkaline solution making it’s DNA stringy
Gram positive cells will not lyse

Microbiology (6 decks)

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