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185 Cards in this Set
- Front
- Back
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Arm
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Base
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Condenser
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Course and Fine Focus Knobs
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Iris diaphragm
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Light Rheostat and should be set at 4.5
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Light Source
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Light Switch
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Objective Lenses
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Ocular Lenses (10x)
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Stage with Stage Clips
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What power objective lens should you start with for all bacterial slides?
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10x
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What power objective lens should you start with for all eukaryotic slides?
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4x
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Should you always view bacteria slides in oil immersion?
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yes the 100x objective lens
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Define total magnification.
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Objective lens x Ocular lens (10x)
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What is resolution?
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is the amount of detail you can see in an image.
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What is the Sterile Technique?
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allows for the transfer of organisms from one place to another without contamination.
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What is a pure culture?
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Means that only one type of organism is present.
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Define aseptic technique?
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vital in hospital settings in preventing cross contamination; flame sterilization of all instruments and tools all the time.
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What percentage of hospitalized patients acquire healthcare associated infections?
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5 to 15%
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What is
nosocomial infections |
hospital acquired infections
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Why do we perform the streak plate technique?
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to produce isolated colonies of one species of bacteria
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What is this?
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pond water (the algae help identify it since it shows up green)
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Which organisms in pond water are eukaryotic?
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Algae and protozoa
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why is fluorescence microscopy is used
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provides contrast to see specimen glowing a color against a dark background
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Identify the types of cells
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Rod & Coccus
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What type of staining technique is used?
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Gram stain
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What type of staining technique is used?
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Leifson Flagella stain
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What type of staining technique is used?
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Capsule Stain
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What type of staining technique is used?
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Schaeffer-Fulton endospore stain
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what type of staining technique is used?
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Ziehl-Neelsen stain
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What type of stain is the Gram stain?
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A differential stain, 1st stain of choice in identifying an unknown.
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What are the steps of a Gram Stain
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Heat fix smear
Primary stain is applied mordant is added decolorized with ethanol counterstain (safranin) Wash |
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What type of mordant is used in the gram stain?
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Iodine
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What is the primary stain used in the Gram Stain?
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Crystal Violet
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What is the counterstain used in the Gram Stain?
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Safranin (red)
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Name four secondary stains
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Acid-fast
Endospore Flagella Capsule |
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What type of technique is used in the Acid Fast Stain?
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Ziehl Neelsen
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What is the primary stain used in the Ziehl Neelsen technique?
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Carbolfuschin (pink)
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What is the counterstain used in the Ziehl Neelsen technique?
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Methylene blue
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Which two techniques are used for endospore stains?
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1 - crystal violet method
2 - schaeffer-fulton method |
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Describe the crystal violet method
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negative staining technique
stains vegetative cells highlighting endospores in purple |
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describe the shaeffer fulton method
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uses a primary stain malachite green to stain endospores
rinse counter stain safranin |
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What technique is used for the flagella stain
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Leifson's flagella
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Describe Leifson's flagella
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rolling drop slide
air dried stain (contains a mordant) applied -inflate flagella |
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What is the special external protective layer that makes viewing capsules possible?
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defined by the form of glycocalyx; an external outer protective layer
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Define the capsule stain
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negative stain
uses India Ink to stain background Counterstain Safranin to stain bacterial cells air dried; heat fixing gives false results |
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What is brownian motion?
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vibration effect
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how do you determine motility?
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Using a hanging drop technique
purposeful forward swimming |
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Motility: Is it motile and what is it's shape?
Bacillus subtilis |
M+, big rod
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Motility: Is it motile and what is it's shape?
Enterobacter aerogenes |
M+, rod
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Motility: Is it motile and what is it's shape?
Klebsiella pnemoniae |
M-, rod
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Motility: Is it motile and what is it's shape?
Micrococcus luteus |
M-, coccus
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Motility: Is it motile and what is it's shape?
Proteus Mirabilis |
M+, rod
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Motility: Is it motile and what is it's shape?
Rhodospirillum rubrum |
M+, rod
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Motility: Is it motile and what is it's shape?
Sporosarcina ureae |
M+, coccus
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Motility: Is it motile and what is it's shape?
Janthinobacterium lividum |
M+, rod
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What is the Gram Rxn & Morphology for:
Bacillus Subtilis (agar) |
G+, rod
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What is the Gram Rxn & Morphology for:
Bacillius Subtilis (broth) |
G+, rod
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What is the Gram Rxn & Morphology for:
E.Coli (agar) |
Gneg, rod
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What is the Gram Rxn & Morphology for:
E. Coli (broth) |
Gneg, rod
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Identify the bacteria, Gram reaction and morphology.
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Bacillus subtilis
G+ Rod |
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Identify the bacteria, gram reaction, and morphology.
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E. Coli
Gneg Rod |
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Identify the bacterium based on colony morphology and it's optimal temperature.
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Bacillus Globigii
37*C |
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Identify a bacterium based on colony morphology and its optimum temperature.
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Bacillus Licheniformis
37*C |
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Identify a bacterium based on colony morphology and its optimum temperature.
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Bacillus Megaterium
Not much difference b/w temperatures. |
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Identify a bacterium based on colony morphology and its optimum temperature.
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Bacillus Mycoides
22*C Fungal/Mold like growth |
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Identify the bacterium based on pigmentation and note any specifics re: temperature.
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Bacillus globigii
No noticeable differences b/w temperatures. |
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Identify the bacterium based on pigmentation and note any specifics re: temperature.
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Chromobacterium violaceum
slightly darker at 22*C |
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Identify the bacterium based on pigmentation and note any specifics re: temperature.
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Micrococcus luteus
bright yellow at both 22 and 37*C |
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Identify the bacterium based on pigmentation and note any specifics re: temperature.
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Micrococcus roseus
salmon pink only grows at 22*C |
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Identify the bacterium based on pigmentation and note any specifics re: temperature.
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Pseudomonas aeruginosa
opaque yellow at 22 watery blue at 37 |
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Identify the bacterium based on pigmentation and note any specifics re: temperature.
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Serratia marcescens
Red at 22*C |
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Identify the bacterium based on pigmentation and note any specifics re: temperature.
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Janthinobacterium lividum
dark purple at 22*C NG at 37*C |
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Why do non-photosynthetic bacteria produce pigments?
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as byproducts of metabolism, as antioxidants, and as protection against solar radiation.
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Name the slime mold that we used in lab and the medium used.
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Physarum polycephalum sclerotia
Tap water agar plates (2%) |
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Define cytoplasmic streaming.
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The flow of oxygen and nutrients from one part of the plasmodium to another in protoplasmic vessels.
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Define hanging drop slide
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suspends bacteria in a drop of fluid, reducing evaporation and water currents when compared to regular wet mounts.
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Define osmotic pressure
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indicates the solute concentration of the surrounding solutions and the effect it has on the flow of water across the membrane.
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What is the term to describe water leaving a cell that results in cell dehydration and shrinkage?
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plasmolysis
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Identify the test and the bacterium had the widest range of salt tolerance and explain why. Which bacterium is an example of an obligate halophile.
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Osmotic Pressure Test
Widest Range: Staphylococcus epidermidis (up to 15% NaCl) because it lives in salty human skin Obligate Halophile: Halobacterium salinarium |
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True or False: Bacillus stearothermophilus is an example of a thermophile.
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Bacillus stearothermophilus
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Name the five temperature classifications for organisms.
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Psychrophiles "cold loving"
Psychrotrophs Mesophiles "middle loving" Thermophiles "heat loving" Hyperthermophiles/Extreme thermophiles |
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What it the temperature range for psychrophiles?
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-5*C and <19/20*C
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What it the temperature range for psychrotrophs?
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-1*C and 35*C
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What it the temperature range for Mesophiles?
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12*C to 45*C
optimum 37*C |
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What it the temperature range for thermophiles?
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41*C and 80*C
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What it the temperature range for Hyperthermophile?
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75*C and 112*C
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Which is room temperature:
37*C or 22*C |
22*C
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Which is body temperature:
37*C or 22*C |
37*C
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Define
bacteriostatic |
subsequent growth after initial incubation
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Define
bacteriocidal |
lack of growth after initial incubation
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What are two examples of bacteriocidal methods
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Cooking and boiling
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What are two examples of bacteriostatic methods
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Refrigerating and freezing
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What is the classification, optimum temp and temp range for
Bacillus licheniformis |
Mesophile
Optimum 37*C Range: 12*C to 45*C |
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What is the classification, optimum temp and temp range for
Bacillus stearothermophilus |
Thermophile
Optimum: 55*C Range: 41*C and 80*C |
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What is the classification, optimum temp and temp range for
E.Coli |
Mesophile
Optimum: 37*C Range: 12*C to 45*C |
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What is the classification, optimum temp and temp range for
Janithinobacterium lividum |
Psychotroph
Optimum 22*C Range: -1*C and 35*C |
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If a bacterium grows at a warmer temperature after incubating in a colder temperature - it is...
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bacteriostatic
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If a bacterium fails to grow at a warmer temperature after incubating in a colder temperature - it is...
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bacteriocidal
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What baceterium is an example of an anaerobe?
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Clostridum sporogenes
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Define obligate aerobes
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require oxygen, possess enzymes, catalase and superoxide dismutase (SOD)
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What is SOD
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superoxide dismutase
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What does catalase breakdown?
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hydrogen peroxide into O2 and H2O
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Define anaerobes
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cannot use oxygen in metabolic pathways
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Define
microaerophiles |
require only small amounts of O2 use aerobic respiration for energy
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Define
Obligate anaerobes |
only grow when no O2 is present; use fermentation or anaerobic respiration for energy
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Define
Facultative anaerobes |
can grow with or without O2; but grow best if O2 is present; possess enzymes, catalase and SOD
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Define
Aerotolerant anaerobes |
primarily fermenters; possess enzyme and SOD
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What does SOD breakdown?
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breaks down superoxide radical into hydrogen peroxide
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Which bacterium is an example of an obligate aerobe?
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Bacillus subtilis
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Which bacterium is an example of a strict anaerobe?
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Clostridium sporogenes
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Which bacterium is an example of a microaerophile and how can you tell?
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Lactococcus lactis
best growth in reduced O2 when compared to AER tubes |
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Which bacteria are facultative anaerobes?
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Enterobacter aerogenes & E.Coli
They will grow most near the top but appear throughout the tube |
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Which bacterium is an example of aerotolerant anaerobe?
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Lactobacillus casiae
will look as though growing from bottom up |
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Describe how UV radiation controls bacterial growth.
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UV radiation kills microorganisms by damaging DNA. It creates thymine dimers that distort DNA, which lead to replication and transcription problems.
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Do bacteria have light and dark repair enzymes?
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Yes but these can be overwhelmed if exposure is significant.
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Which type of environments are most susceptible to UV radiation?
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Microrganisms in air, clear water, and on surfaces.
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How do hospitals use UV radiation?
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In germicidal lamps (ORs and nurseries).
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Differentiate a disinfectant from an antiseptic.
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Disinfectant is generally a chemical that kills or inhibits vegetative microbes. An ANTISEPTIC is a type of disinfectant that is nontoxic enough to be applies to skin and mucous membranes.
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Is this an antiseptic or disinfectant?
Isopropyl Alcohol (IA) |
Antiseptic
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Is this an antiseptic or disinfectant?
Hydrogen Peroxide (HP) |
Antiseptic
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Is this an antiseptic or disinfectant?
Lysol (Ly) |
Disinfectant
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Is this an antiseptic or disinfectant?
Bleach (Bl) |
Disinfectant
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What does a zone of inhibition indicate?
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antimicrobial activity
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You are looking at Bacillus Subtilis. Which chemical was LEAST effective and is it a disinfectant or an antiseptic?
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Isopropyl alcohol
antiseptic |
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Identify this bacterium. Hint: it was the only one that had NG for HP and Bl and it is "topical".
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Staphylococcus epidermidis
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Which of these bacterial species is most UV resistant/tolerant and why?
Bacillus Subtilis, E. Coli, Staphylococcus epidermidis |
Bacillus subtilis; endospores resist UV;
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You are traveling when you suddenly need surgery. The village hospital only has liquid hand soap available for their doctors to scrub in. You have a small bottle of alcohol-based hand sanitizer in your day pack.
Do you... A) go with the liquid based soap B) insist on the alcohol based hand sanitizer EXPLAIN WHY |
B - you insist on the alcohol-based hand sanitizer because
-it is more effective at killing transient organisms and agents -it reduces the number of hospital acquired infections -and you saw in lab that it killed 93% of bacteria present on your partner's hands. |
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True or False:
Molds belong to the Kingdom Plantae? |
False. Molds belong to the Kingdom Fungi
Molds include mushrooms, mildew, and bread mold. |
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True or False:
Yeasts belong to the Kingdom Bacteria? |
False. Yeasts belong to the Kingdom Fungi include unicellular nonmotile yeasts.
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Identify these life stages and identify type of mold.
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A. Spores
B. Spore germinating hypha C. Hyphae going to Mycelium Aspergillus Mold |
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Name this mold.
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Rhizopus
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Name this mold.
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Penicillium
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What was the name of the selective medium used in the mold lab?
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Sabouraud Agar due to its low pH and high sugar concentration which inhibits bacterial growth
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What stain was used to make the molds visible?
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lactophenol cotton blue
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How did we collect the mold?
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Tape and toothpicks.
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What was the name of the yeast used in lab?
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Saccharomyces cerevisiae
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What are you looking at?
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Budding Yeast is most clearly present
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What is it?
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Yeast ascospore
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What method is used to estimate the number of bacteria in the soil.
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serial dilution of a sample of soil.
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Define serial dilution
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dilute one gram of sample soil through a series so that bacterial numbers are progressively decreased
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Why, when using the serial dilution method, are estimates low?
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because the artificial and limited growing conditions are not suitable for all organisms.
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You CFU count for soil bacteria is 88 on a 10*-6 (power) plate. Is the CFU of original gram sample:
A. 88 CFUs B. 88 x 10*-6 CFUs C. 8.8 x 10*7 CFUs |
8.8 x 10*7 (power) of CFUs
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Define coliforms
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all aerobic and facultative anaerobic Gneg rods which produce acid and gas from lactose fermentation in 48 hours at 35*C
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Coliforms
AKA |
indicator organisms
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What is the 2nd step of water testing?
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Confirmed Test
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What are the three steps of water testing (in order)?
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Presumptive Test
Confirmed Test Completed Test |
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What constitutes a positive presumptive test (possible coliforms)?
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both acid and gas present
(yellow and bubble in durham tube) |
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In a presumptive test what do yellow and blue tubes with no bubbles indicate?
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Yellow indicates presence of acid and blue means no acid was produced; no bubble means no lactose fermentation.
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How do you calculate the MPN of a presumptive test?
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Count the number of positive big tubes; look at chart; count the number of positive baby tubes; look at chart; arrive at answer.
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Why must the confirmed test be performed for water testing?
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it is necessary due to the synergism of the mixed bacteria in a tube; giving a false positive.
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In the confirmed tests (water) name the differential and selective mediums used.
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Differential medium: EMB Eosin Methylene Blue
Selective medium: EMB Eosin Methylene Blue |
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Describe Eosin Methylene Blue as a differential medium
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a differential medium because it contains lactose and dyes - it differentiates non-coliforms by dying coliform colonies
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Describe Eosin Methylene Blue as a selective medium
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As a selective medium EMB selects Gram negative bacteria
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why does a green sheen appear on E. coli. EMB agar plates?
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The acids cause a reduction in pH and precipitate the dye complex (methylene blue eosinate) "dye sink" onto the colonies
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Identify they areas as indicated.
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A. E.Coli due to green sheen
B.Enterobacter aerogenes due to pink colonies C. Non-Coliforms colorless colonies that transmit the reddish-purple color of the medium D. Enterobacter aerogenes forming purple colonies (note no green sheen) |
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What is a parasite?
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An organism that lives on or in another organism causing harm.
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Describe ectoparasites
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Ectoparasites live on the host
Complex life cycle multiple morphologies sexual and asexual reproduction multiple hosts protozoa - single celled motile |
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Describe eukaryotic parasites
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the human serves as a reservoir of infection
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What are the major ways that parasites are spread?
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Via food, water, vectors, soil
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What is a vector?
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Organism that transmits an organism from one stage to another
e.g., Anthropods (insects) |
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What is an intermediate host?
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Intermediate Host provides a living space where parasites asexually reproduce
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What are some ways that scientists are using to try and stop/slow the spread of parasites.
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Protective measures for food and water supply
Filtration of water and access to clean water |
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can parasites ever be completely eliminated?
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Uncontrollable and they can infect multiple hosts that can infect humans
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Identify the picture.
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Plasmodium Falciparum (Malaria infection)
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Identify the picture
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Ascaris lumbricoides eggs
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Name the three categories of Helminths.
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Nematodes - round worms
Cestodes - tapeworms Trematodes - Flukes |
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Which category contains Ascaris lumbricoides:
Nematodes, Cestodes, Trematodes |
Nematodes
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How is malaria vectored?
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Via the female mosquito Anopheles
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Name some malaria prevention methods.
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Prophylactic medications
prevention of mosquito bites mosquito eradication (insecticides) |
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List some diseases vectored by mosquitos.
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Malaria
filariases dengue fever yellow fever west nile virus |
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list some diseases vectored by ticks
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lyme disease
rocky mountain spotted fever ehrlichiosis |
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If you have a starch agar medium and there is a clear area around the growth line of a bacteria; what test are you looking at?
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Amylase Test
Positive = clear area around bacteria growth line iodine reagent used to visualize exoenzymes present |
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If you have a tributyrin agar medium and there is a clear area around the growth line of a bacteria; what test are you looking at?
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Lipase
Positive = clear area around bacteria growth line |
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If you have a gelatin agar medium and there is a clear area around the growth line of a bacteria; what test are you looking at?
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Gelatinase
Positive = clear area around bacteria growth line |
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Of the exoenzyme test what was the trend?
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Bacillus subtilis tested positive for exoenzymes across all three.
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Describe Catalase test
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Tests for the respiration enzyme catalase using substrate hydrogen peroxide
Positive = bubble formation |
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Describe oxidase test
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tests for the respiration enzyme cytochrome c oxidase
positive = purple spot |
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Describe the nitrate reduction test 1
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tests for the ability to reduce nitrate to nitrite using nitrate reductase in potassium nitrate broth
positive = bubbles present |
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Describe nitrate reduction test 2
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tests for nitrite using reagents sulfuric acid and Tromsdorff's reagent
Positive = immediate purple brown |
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Describe nitrate reduction test 2
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testing for nitrate
zinc added to negative nitrite tests positive = purple brown negative = reduction to another gas like ammonia possible |
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For the nitrate reduction tests, what were the two extremes?
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Bacillus megaterium immediate results;
Pseudomonas aeruginosa N2 gas positive only |
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For the carbohydrate fermentation tests what were the three sugars tested for?
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Glucose, Lactose, Sucrose
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Of the carbohydrate tests, what were the two extremes?
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Alcaligenes faecalis grew on peptones for all three GLS
E.Coli the only one with AG for all three sugars |