BIOL MicroLAB Midterm

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genus then species, name is italicized, or underlined. Genus species

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1

genus then species, name is italicized, or underlined. Genus species

How to write a binomial name

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2

white paper biohazard discard pouches taped to the bench or the red plastic biohazard boxes

where to discard microscope slides, broken glass

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3

orange/red biohazard trash bags in the white metal cans with foot pedal to open

discard petri dishes, plastic tubes, paper trash contaminated with microbial culture

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4

plastic discard tubs on the bottom of the cart, use racks for tubes, remove all labels

discard tube and flask media

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5

plastic discard beakers on the bench tops or in the orange/red biohazard trash bags in the white metal cans with foot pedal to open

discard micropipette tips, swabs, tongue depressors

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6

long plastic discard tubs on the bench tops provided only when needed

discard glass serological pipettes

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tall boxes labeled for glove discard only

discard gloves

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8

regular trash cans

discard non-contaminated paper trash

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9

do not discard; return to the top of the cart

discard unused media

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10

culture medium

contains nutrients, like carbon and nitrogen, for cells to grow

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11

complex medium

composed of digests of chemically undefined substances such as year and meat extracts or digests

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defined medium

a precise chemical composition is known

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broth

liquid medium

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agar

solid media

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plates

solid media in petri dishes

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deeps

test tubes containing solid medium, which has been allowed to solidify in an upright position, used mainly for anaerobic growth

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slants

tests tubes containing solid medium that solidified at an angle

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selective medium

medium to favor the growth of certain microbes and to inhibit undesirable competitors

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differential medium

a medium that provides a visible indication of a physiological characteristic of a microorganism

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sterilization

killing or removal of all living organisms and their viruses from a growth medium like autoclaving

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autoclave

sealed device that allows the entrance of steam under pressure

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dry heat sterilization

kills by oxidation effects

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flaming

one of the simplest methods of dry heat sterilization

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hot-air sterilization

sterilizing glassware in an oven

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filtration

heat-sensitive liquids or gases are sterilized

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gas chemosterilizers

sterilization for heat sensitive objects like a plastic petri dish

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ethylene oxide

widely used gas for sterilization

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cold-sterilization

no heat is used for sterilization

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electromagnetic radiation

microwaves, ultraviolet radiation, X-rays, gamma rays, and electrons for sterilization

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ultraviolet radiation

causes damage to DNA, leading to the death of exposed organisms, cannot penetrate solid and is only useful to sterlize exposed surfaces

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ionizing radiation

causes ions and other reactive molecules to be produced and these reactive molecules can degrade or alter biopolymers such as DNA and proteins

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contaminants

unwanted organisms

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33

asceptic technique

used to prevent contamination of bacterial cultures and the environment

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34

shake culture tube, heat loop, remove cap, flame neck of tube, cool loop, take a loop of microorganisms, flame mouth of tube, return cap

process of asceptic technique

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35

oil-immersion objective

used to observe the cells at 1000x

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36

cells mostly consist of water, so unstained cells may be difficult to see bc of the lack of contrast

why is the oil needed to see at 1000x?

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37

staining cells makes them more visible

how do you make cells more visible?

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38

smear

first step to staining cells, only made from cultures in liquid or on solid medium, a thin layer of cells on a slide

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heat fixation

kills the cells, destroying autolytic enzymes, helps cells adhere to slide

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prevents the cells from being washed from the slide during staining

Why is adhesion of cells to the slide important?

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41

morphology

shape of cells

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42

coccus, rod/bacillus, and spiral

three types of basic shapes

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diplo (pairs), strepto (chains), staphylo (clusters)

three ways cells are arranged

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44

cocci

round shape

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bacilli

rod-shaped

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spiral

twists

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prosthecate cells

organisms that produce prosthecae

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48

prosthecae

cellular appendages that increase cell surface area, which help transport nutrients into the cell

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filamentous growth

some actinomycete bacteria, produce filaments or hyphae that form a mycelium

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mycelium

network of hyphae

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simple stain

a direct stain that uses an aqueous or alcohol solution of a single basic dye, makes cellular shapes and structures visible

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negative stain

an indirect stain that uses an acidic dye to color the background, leaving the cells colorless

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differential stain

utilize a combination of dyes to demonstrate a chemical or structural component of a cell

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Gram-positive and Gram-negative (the gram should always have an uppercase G)

two groups of gram stains

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peptidoglycan

repeating disaccharides attached by a polypeptide to form a lattice in the cell walls

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Gram-positive has many layers of peptidoglycan and a rigid structure, also contains teichoic acids; Gram-negative has less layers, lipopolysacchardies (LPS) outermembrane, no teichoic acid

How does Gram-positive cells differ from Gram-negative?

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teichoic acids

copolymers of glycerol phosphate and ribitol phosphate and carbohydrates linked by phosphodiester bonds that provide the wall of antigenic specifity

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primary stain

for a gram stain, its crystal violet

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mordant

iodine in a gram stain, binding to crystal violet to form a not easily removed complex

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decolorizer

ethanol in a gram stain, removing the basic dyes like crystal violet

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counterstain

safranin in a gram stain

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Gram-positive staphylococcus epidermis, gram-negative purple spirals

When viewing a gram stain under 100x what do you see?

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younger, less than 24 hours since older might have a cell wall structure that may be compromised, causing cells to lose the ability to retain the primary stain

Is gram negative more consistent on young or older bacteria?

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purple

What color are gram-positive?

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red

What color is gram-negative?

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endospore

dormant structure that is commonly produced by certain Gram-positive rods, forms when nutrients or water is depleted, highly durable dehydrated metabolically dormant structures

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they are so resistant, their survival could produce toxins and cause disease

Why are endospores dangerous?

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Geobacillus stearothermophilus

bacterial endospores that are used to indicate the effectiveness of autoclaves

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vegetative cell

one that can grow and divide under optimal conditions

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cryptobiotic state

endospores are in this state; no metabolic activities are occurring

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sporulation

when nutrients or water become depleted, this process will begin coverting the vegetative cell into a single endospore; process of endospore formation within a vegetative cell

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spore septum

isolates the replicated chromosome and a small amount of cytoplasm in the first stage of endospore formation

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forespore

structure that is a double-layered plasma membrane surrounding the genetic material

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cortex

layer where peptidoglycan will be laid down between the two plasma membranes

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spore coat

a coat around the outside of the structure formed by spore proteins; responsible for the resistance of the endospore to harsh chemicals

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core

consists of the cell wall, cytoplasmic membrane, cytoplasm, and nucleoid

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central, terminal, or subterminal positition

Where can endospores be located?

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small acid-soluble proteins (SASPS) and calcium-complexed dipicolinic acid

protecting the endospore DNA from damage

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outgrowth

the synthesis of DNA, RNA, and bacterial proteins

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malachite green

Primary stain in endospore stain

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safranin

Counterstain in endospore stain

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mycobacterium

What culture is used in acid-fast stain?

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mycolic acid

makes mycobacterium cells hard to stain; resists decolorization

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acid-fast

since mycobacterium resist decoloration they are classified as?

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hydrophobic nature

reason for the wrinkled appearance of mycobacterium; make cells impermeable; slow growth of mycobacterium

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egg albumin

help dispense the mycobaterial cells when making a smear; high protein content; help make smear for acid-fast stain

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carbolfuchsin (magenta dye)

primary stain for acid-fast solutions

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acid-alcohol

removes fuchsin dye from cells that are not acid-fast

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methylene blue

counterstain for acid-fast reactions

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blue

What color are cells that are not acid-fast?

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91

capsule

contains layers of polysaccharide that surrounds the cell wall of bacteria

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Azotobacter vinelandii

What is one of the bacteria that produces a polysaccharide capsule?

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presence of excess nutrient

What do capsules need to be formed?

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desiccation of cell

water prevents this of the cell

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virulence factor

if a capsule enhances the organism’s ability to cause disease

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avirulent

nonencapsulated form that does not cause disease

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congo red

primary stain for capsule stain

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Maneval’s stain

darkens the background and stains cells in capsule stain

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99

background is gray to darkblue/purple, cells are red, capsules are unstain and appear clear around the cell

What color is the background and cells in a capsule stain?

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100

to isolate and maintain a microbial culture

What are steak plates for?

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