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195 Cards in this Set
- Front
- Back
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What are the 3 dry objectives?
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They are the 4x, 10x, and 45x
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Which objectives lens is the immersion objective?
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100x and this is the preferred lens for use in Microbiology for examining bacteria
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What is resolution?
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It is the ability of the lens to distinguish fine detail
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Define parfocal
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Lenses of the microscope are adjusted so that the specimen remains almost in focus when a higher magnification objective is moved into position.
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Where are the oculars and what do they do?
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The eye piece we look through is the ocular and it effects the final magnification of the specimen. they are adjustable and magnify 10x
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What is the body tube?
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It is the intermediate magnification lens located between the oculars and the Objectives
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What is the revolving nosepiece?
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Places a given objective in the viewing position
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What is the mechanical stage?
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Holds the specimen slide and controls movement of the slide horizontally and vertically in four directions
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What are the objectives?
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The source of initial magnification of the specimen. Transmits a magnified and inverted specimen image to the oculars.
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What is the condenser?
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Focuses light on the specimen and through the objective
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What does the iris diaphragm do?
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It decreases glare by regulating the amount of light entering a given objective
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What does the condenser objective knob do?
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raises or lowers the condenser
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What does the course focus knob do?
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Raises or lowers the stage (or the objectives, depending on manufacture design). Used for INITIAL focus upon the specimen
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What does the Fine focus knob do?
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When adjusted continuously during specimen examination, obtains optimum sharpness of the speciman image.
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What does the green or blue filter do?
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It absorbs excess red rays from filament lamp when placed on the light source.
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What objective do we start with in Microbiology?
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10x
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What is the purpose of immersion oil?
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To increase resolution by funneling the light to the oculars
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How do you calculate final magnification?
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Ocular Magnification times the Objective magnification
ex: 10x objective times 100x Ocular equals 1000x magnification |
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What are three ways to affect the contrast of a specimen?
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1) raise or lower condenser
2) adjust light by opening or closing the iris diaphragm 3) Turn the rheostat / light intensity switch on or off |
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For stained slides, what is usually the best position for the condenser?
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In the raised position because more light is needed.
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What is the free working distance and for which objective is it greater?
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It is the space between stage and objective and is greater for the lower objectives
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Magnification is expressed in ________ and is designated by _____
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diameters, X
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The larger the utilized cone of illumination, the __________ the resolution
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Greater
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In comparing a 10x versus a 100x objective, which has a greater utilized cone of illumination?
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100x (about average of 14 degrees versus 110 degrees
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List 3 things you need to do for shutting down the microscope
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1) Turn objective to lowest power and move the stage and objectives apart.
2) Wipe all lenses clean with lens cleaner and soft tissue/cloth. 3) cover microscope with dust cover and put in cabinet |
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After putting the specimen on the slide, what is the next step before applying the Gram stain reagents?
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Heat fix the specimen
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What are 3 reasons for heat fixing a slide?
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1) It kills the bacteria
2) Dead bacteria stain better 3) It fixes/adheres stain to the slide |
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What are the four steps of making a Gram stained slide after it has been heat fixed?
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1) Apply Crystal Violet for 1 minute
2) Apply Gram's Iodine for 1 minute 3) Apply decolorizer till the RUNOFF is clear 4) Apply safranin for 1 minute |
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What does Gram's iodine do?
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It helps the Crystal violet set
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Does decolorizer ALWAYS remove the color on a bacterial Gram stain?
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No, it may or may not decolorize and amount of decolorizing will vary
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Decolorizer is what type of solvent?
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acetone/alcohol solvent
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Safranin is what color and is also called a ____________ stain
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Pink, counter
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Who was the Noble prize winning scientist who first proved that bacteria cause disease?
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Robert Koch
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Koch discovered __________ shaped bacteria in blood of animals that had died of anthrax, took that blood and injected it into healthy animals who then _______________________
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ROD, died of anthrax and had the same ROD shaped bacteria in their blood
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What are the 5 kingdoms of living things?
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Plantae
Animalia Protista Fungi Procartyotae |
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What is a Eukaryotic cell?
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Cell has a true nucleus that contains genetic material and a nuclear membrane separating it from the cytoplasm
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What is a Prokaryotic cell?
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A cell that lacks a true nucleus, DNA/RNA float in the cytoplasm
Bacteria belongs here |
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Why are viruses not considered a living thing?
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Because they require a living HOST cell to survive
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What are 2 characteristics of the Plantae kingdom of living things?
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Have Eukaryotic cells and are multicellular
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What are 2 characteristics of the Animalia kingdom of living things?
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Have a Eukaryotic cells and are multicellular
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What are 2 characteristics of the Protista kingdom of living things?
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Have Eukaryotic cells and mostly UNIcellular
ex: Giardia Protozoan |
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What are 2 characteristics of the fungi kingdom of living things and what are 2 examples?
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Have Eukaryotic cells and may be uni OR multicellular
Ex: Yeasts and molds-Ringworm is an example of a mold |
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What are 2 characteristics of the Procaryotae kingdom of living things?
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Have PROKARYOTIC cells and are unicellular
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When naming microorganisms, the Genus is:
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Listed first
Always capitalized Underlined or italicized (is like a last name....) |
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A Genus is:
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A number of related species
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A species is:
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Each different type of organism within a Genus
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When naming microorganisms, the species is:
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Underlined or italicized
Started with a small letter (is like a first name....) |
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Gram positive bacteria stain what color?
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PURPLE
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Gram negative bacteria stain what color?
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PINK
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Gram positive bacteria stain purple because:
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Decolorizer only affects the lipid layer only on these cells because the large peptidoglycan layer of cell wall protects the cell from the chemicals in the decolorizer
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Why do Gram negative cells get decolorized?
Adding __________ makes them stain pink? |
Because they have a thin layer of petidoglycan which allows the decolorizer into the cell because the large layer of lipopolysacchride does not protect the call and is broken down.
Safranin |
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Animal cells/Human cells have ________ but NOT _____________ and Bacteria have _____________
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Cell membranes
Cell walls BOTH |
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Cell wall is a __________ outer layer that does what?
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RIGID,
Provides shape Protects cell and its contents Determines Gram reaction |
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What are 3 shapes of bacteria?
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cocci- spherical/round ex: streptococci
Rods-Bacillus, coccobacillus (oval shaped) ex: Bacillus antracis-anthrax Spirals-spirochetes ex: leptospira |
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Cell membrane is _______ ______-permeable.
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selectively semi-
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The cytoplasm of Bacterial cells contains:
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Nutrients, electrolytes, H20, DNA, RNA, ribosomes for protein synthesis
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What is flagella?
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A long thin whip-like appendage for motility that some bacteria may have and may aid in the spread of disease
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What is a capsule?
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A gelatinous, slimey, sticky outer cell surface that prevents phagocytosis.
Only some bacteria have this and those that do are often involved in dental disease. |
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Name a bacteria that has flagella, causes diarrhea in animals and humans and zips around under the microscope when a wet mount micro exam is done
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Campylobacter
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What are Pili?
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Hair like projections that aid in adherence to mucous membrances and only some bacteria have them
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What is a spore (endospore)?
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This is a highly resistant stage that occurs during times when the environmental conditions are no longer favorable for growth. A dormant stage that bacteria are capable of staying in for a long time until conditions are favorable.
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Although spores are very strong and difficult to kill, what are 2 effective ways to get kill them?
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Bleach
proper autoclaving techniques |
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What is a vegetative cell?
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a cell that is metabolizing and reproducing (versus a dormant endospore cell)
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What common URI is caused by a bacteria that has Pili that make it difficult for the MCTS to push it out?
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Bordatella
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What is the MCTS?
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The Mucociliary Transport System
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What are 3 spore forming bacteria?
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1) Clostridium tetani -Tetnus, lockjaw
Humans and horse most susceptible, anaerobic, spores in dirt 2) Clostridium botulinum- Cuases Botulism, Anaerobic 3) Bacillus anthracis- anthrax |
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Bacteria replicate using:
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Binary fission
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Bacteria can be grown on/in many types artificial media, one of which is called agar and should be incubated at ____________ for at least _____hrs
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35-37 degrees Celcius/ ~99 degrees F
24hrs |
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What are the basic elements needed for all living cells?
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Carbon
Hydrogen Oxygen Nitrogen Sulfer Phosphorus ---which are all part of the carbohydrates, proteins, lipids and nucleic acids of cell requirements along with K-Potassium, Ca-calcium, Mg-Magnesium, Fe-Iron |
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What are 6 ways Normal Flora (NF) bacteria help the host?
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1) Produce vitamins in intestine, especially vitamin K
2) Metabolism of drugs 3) Excretion of bilirubin which results from the breakdown of hemoglobin from dead RBC's 4) Prevention of disease 5) Digestion in ruminants and horses 6) Used to produce drugs such as insulin |
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What are 5 places in the body that are sterile and should NOT have bacteria, even Normal Flora?
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1) Internal Organs
2) Blood 3) CSF-- meningitis results when bacteria invade this 4) Synovial fluid- Lyme disease is one area where this body part is invaded 5) Urine IN the bladder |
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What is it called when bacteria invade the blood?
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Bacteremia
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What is it called when bacteria are found in the blood and organs?
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Septicemia
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Pathogens are__________ and there are ______ types
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Disease producing microorganisms, 2
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What are the 2 types of pathogens?
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Opportunistic
Obligate |
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What is an opportunistic pathogen and when are 2 times they usually invade?
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They are bacteria that do not normally cause disease but will given the right opportunity such as a host that is compromised or bacteria in an unusual location.
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Bacteria that are normal flora in one part of the body cause infection when in the wrong location. For example, E. Coli is normally found in the intestine but when it is found in the:
Bladder it causes ___________ Uterus it causes___________ mammary glands it causes ______ Abdomen it causes ____________ |
Cystitis
Pyometra Mastitis Peritonitis |
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What are some factors that cause a host to be compromised?
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--Very young -immature immune system
--very old -immune function is deteriorating --Underlying disease --Antibiotics, steroids --Viral infections such as HIV or FIV --IV or urinary catheters --Wounds, lacerations and incisions |
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Long term use of steroids can ______
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delay healing and are also immunosuppressive, therefore making host more prone to infections
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If an obligate pathogen is present in an animal, the animal _____________
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WILL usually get the
disease |
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Obligate pathogens existence depends on disease production and _______________
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Ability to spread
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Examples of Obligate pathogenic diseases are:
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Rabies
Anthrax Brucellosis (Bang's Disease) Plague |
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What is Epidemiology?
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The study of disease transmission causes, distribution as well as control and prevention
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Define incubation period
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The time from initial infection until clinical signs appear. Animal may appear healthy but may be infectious, contagious.
Bacteria typically are 2-5 days, Viruses 3 days to 2 weeks |
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What is a nosocomial infection?
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Hospital acquired infection that occurs in approx 5% of patients
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What is zoonosis?
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Transmission of disease from animals to people
ex: TB, brucellosis, rabies, plague |
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What is inverse zoonosis?
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Transmission of disease from people to animals. TB can be spread from people to some primates
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What is a vector?
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An invertebrate insect that can transmit disease though its bite
ex: Mosquitoes transmit Heartworm, West Nile, WEE, VEE Ticks spread Lyme disease Fleas spread plague |
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What is a fomite?
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An inanimate object contaminated with disease causing microorganisms
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Define endemic
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When an organism prevails continuously in a certain geographic location
ex: Plague is endemic in Colorado and Lyme disease is endemic in the midwest |
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What is an epidemic?
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A sudden outbreak or increase in number of disease cases
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What is a disease reservoir?
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a permanent place for an organism to reside in order to exist and spread to continue its existence .
Can be human, animal, plant, soil, water. Rodents are the reservoir for plague. |
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What is a carrier?
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An infected individual who is a potential source of infection for others and who may or may not be symptomatic
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Disease spreading organisms are spread by these typical routes:
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1) URI by aerosols
2) GI by feces and vomit, EX: Parvo 3) GU -urine, uterine/ vaginal secretions, semen 4) Skin-contact, discharged pus 5) Bodily fluids in general such as contact with blood, saliva, milk |
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True or False
A diseased person/animal is more infectious when they are showing signs of a disease |
TRUE
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What are some ways that disease causing organisms enter a susceptible host?
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~Inhalation such as in Kennel Cough
~Ingestion such as in Parvo ~Inoculation (vector bite)or infection through skin and mucous membrane ~Urogenital tracts (Bang's disease though breeding or unsterile AI equip) ~Transplacental- FIV and Panleuk ~Umbilicus |
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Some ways to decrease the transmission of infectious agents:
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~Best way is to WASH HANDS!!!!
~Use gloves ~Good housekeeping practices ~Sterilization of instruments and use of sterile techniques ~Use of isolation |
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What are the 2 basic mechanisms of disease production for pathogenic bacteria?
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1) tissue invasion
2) Toxin production |
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Bacteria all tend to have an affinity for invading certain types of tissues and their invasion ________________
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Damages/destroys cells and tissues resulting in disease.
EX: Brucella prefers the reproductive tract and cause abortions Strep. pyogenes prefers tissues in the throat and tonsils |
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What are a few items that assist a bacteria in invading tissues and causing disease?
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1) Enzymes destroy tissue
2) capsules prevent phagocytosis 3) Pili aid in sticking to cells to keep bacteria from being expelled 4) Some survive and multiply in phagocytes 5) Leukocidans destroy WBCs |
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What are the 2 types of bacterial toxins?
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Exotoxins and Endotoxins
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What are exotoxins and where are they produced?
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Large protein molecules released by bacteria and are then released into tissues, bloodstream, food
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Exotoxins are toxic to the host in 3 ways:
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Tissue necrosis
RBC hemolysis Neurotoxins |
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2 types of neurotoxin producing bacteria are:
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~~Clostridium tetani -tetanus, which causes very strong muscle contraction
Clostridium botulinum - botulism which causes muscle weakness and paralysis |
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What are some characteristics of Endotoxins?
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~They are a part of the cell wall of GRAM NEGATIVE bacteria, which have a high content of lipopolysacchride
~they are released when Gram negative bacteria die |
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What is Endotoxemia?
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A serious complication of overwhelming gram Negative infections
ex: Bacteremia and Septicemia |
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What is DIC?
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Disseminated Intravascular Coagulation
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Endotoxemia causes production and release of inflammatory mediators which results in:
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-fever
-hypotension -Shock due to pooling of blood in capillaries-so not going where blood is needed -Organ failure -DIC -Coma and death |
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True or false
Skin is a way that a host resists bacterial infection |
TRUE due to it being dry, having an acidic ph and that it constantly sheds
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Mucous Membranes help a host resist bacteria by___________ and some body systems that use this are:
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Trapping bacteria with mucus so it can be phagocytized
1) MCTS of the respiratory system 2) GI Tract with assistance of peristalsis and the HCL which has a ph of 2 because bacteria prefer a ph of 7 3) GU tract- flow of urine |
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What is a macrophage?
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A tissue invading phagocyte that lives in tissues that phagocytizes tissue invaders. Was a WBC monocyte in the bloodstream.
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Normal Flora help a host resist bacteria by ____________________
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Competing with pathogens for sites of attachment, O2 and nutrients
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Phagocytic WBC's and tissue macrophages are 2 parts of what type of host resistance?
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Inflammation and phagocytosis
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The immune response has 2 branches. What are they?
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1. Humoral Immunity with B lymphocytes
2. Cellular Immunity with T lymphocytes |
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Humoral immunity is also known as Humoral Mediated Immunity (HMI) and involves what types of lymphocytes?
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B lymphocytes that recognize antigens (ags) and produce SPECIFIC antibodies (abs)
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What is an antigen?
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a foreign protein (bacteria, virus, bacterial toxin)
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What is an antibody?
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a large protein molecule produced by B lymphyocytes
types: Immunoglobulin (Igg), IgE, IgM, IgE |
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Humoral immunity involves:
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antibodies bind to specific antigens which leads a bacterial or viral antigens being lysed, destroyed and then unable to cause disease.
If antigens are toxins, the antibodies neutralize the toxins |
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B lymphocytes that are in the process of producing a specific abs to a specific ag are called ____________ and this process of making these takes _________ while at the same time other B lymphs become ________
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Plasma cells,
7-10 days Memory cells |
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What are memory B Lymphs?
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Cells that remember same antigen at a later date so if exposed again, the Memory B cells produce antibodies QUICKLY and prevent disease from occurring. This is why IMMUNIZATION works!!!
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Immunization is:
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The injection of antigens that cannot cause disease but will stimulate formation of memory B cells
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What are 3 types of immunizations?
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1) Vaccine- an immunization involving viral antigens
2) Bacterin- immunization involving bacterial antigens 3) Toxoid-immunization containing toxin antigens such as with tetanus |
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Antitoxins, antivenin, immune/gamma globulin are all injections of ________ and are used to ________________
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Antibodies, treat disease or in cases of known exposure, to prevent disease
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Injection of antitoxins and antivenin cause __________ due to no ________ being formed because ________ were injected rather than ________ like in immunizations
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temporary immunity, B Memory cells, antibodies, antigens
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Active immunity is:
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Immunity where antibodies are produced by the host, B memory cells are produced.
Natural is when get disease and recover Artificial is when immunizations are used |
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Passive immunity is:
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Transfer of antibodies and is a temporary immunity
Naturally is through colostrum artificial is through antitoxins, antivenin, injection of immune/gamma globulins |
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What is maternal interference?
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When the antibodies received from the mother interfere with vaccines by binding with the vaccine antigens and end up neutralizing them.
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Cellular immunity aka Cellular Mediated Immunity (CMI) involves what kind of lymphocytes?
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T lymphocytes
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What are the 3 important functions of T cells in the bodies immune response?
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1. Elimination of intracellular organisms such as viruses
2. Elimination of abnormal cells and tissues-involved in cancer and tissue rejection 3. Involved in hypersensitivity reactions such as allergies and anaphlaxis-due to overreaction of super sensitive T cells |
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True of False
CMI works with HMI to fight disease |
True
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True or false
T cells form memory cells just like B cells do? |
true
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True or false
FIV and HIV live within T lymphocytes |
True
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T lymphocytes __________ virally infected cells and _____________.
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recognize
eliminate them |
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Another type of resistance is ________ due to some diseases being __________ specific and individual ____________
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Natural
species ( we do not get Parvo or Feline Leukemia for example) genetics |
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What is purpose of a direct examination of a specimen?
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It is a a microscopic examination of a patient specimen to see if bacteria are present, possibly provide a tentative diagnosis, and lead to appropriate culture media selection.
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Culture swabs contain ___________ media to prevent specimen from ___________ and most bacteria will survive using this transport method for __________ hours.
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liquid
drying out 24-48 |
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Liquid specimens are collected into_______________ container such as a ____________.
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sterile
red top tube or syringe |
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The best way to get a sterile urine sample is________________?
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Cystocentesis
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Dogs and Cats tolerate which procedure better ?
Cystocentesis or catheter placement? |
Cystocentisis is tolerated better. Dogs and cats typically require sedation to be catheterized
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What are 2 other okay ways to collect a urine sample other than cystocentesis?
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Mid-stream catch or urinary cath
Only okay because not as sterile |
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What is the best way to collect a milk sample?
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Mid-stream
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How is CSF collected?
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Through a sterile spinal tap
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What is the name of the procedure for collecting synovial fluid?
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Arthrocentesis
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What is Effusion?
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The accumulation of fluid into a body cavity (pleural, peritoneal, pericardial)
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When blood is collected to check for bacteria, what type of collection is done?
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Serial collection of samples into sterile red top tubes due to the wide amount of variance in bacterial levels
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Tissue specimens are NOT transported in _____________ because it ____________.
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Formalin
kills bacteria |
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Anaerobic bacteria typically cause a wound, diseased and necrotic tissue to __________
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be foul smelling
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Osteomyelitis is ____________ and is typically attributed to _______________ bacteria?
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inflammation of bone and bone marrow
anaerobic |
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Bacteremia is an infection of the ____________ involving _____________ bacteria
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blood
anaerobic |
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What are 4 collection methods used for suspected anaerobic bacteria?
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1) Anaerobic culturette
2) Capped syringe with all air expelled 3) Transport media such as a Port-a-Cul which transports all types of bacteria on swabs -these contain ampules that prevent drying out and to get rid of O2. 4. Thioglycollate broth |
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Media contains ________ used to ___________ .
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nutrients
grow bacteria |
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Agar and broth are 2 types of _______ and it is __________ that causes media to solidify.
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Media
Agar |
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Agar is typically seen in __________ and __________ and bacteria can form _____________ on agar.
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Petri dishes
agar slants (test tubes) colonies |
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What are 5 types of basic Media?
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Basic Nutrient
Enriched Selective Differential/Selective Selective/enrichment |
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Basic nutrient media is a _____________ and will support growth of __________ bacteria and is commonly used because it is ____________.
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general, all purpose media
most cheap |
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Trypticase soy agar(TSA) is a type of _________________
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Basic Nutrient Media
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Enriched media is basic nutrient media with extra______________ to make the media enriched and is a better choice for ________.
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nutrients
fastidious (picky) bacteria |
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What is the most common type of enriched micro media and what is it used to check for?
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blood agar
used to check for degree of hemolysis for assisting in bacterial identification |
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Blood agar (BAP) is enriched with __________ because it their RBC's are very sensitive to hemolysis.
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5% sheep's blood
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Certain types of bacteria produce ___________ which causes ________ of RBC's in blood agar.
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Hemolysins
lysis |
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What are the 3 types of hemolyis?
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Beta
Alpha Gamma |
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Beta hemolysis is indicated by what 2 things?
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COMPLETE lysis of RBC's in blood agar
CLEARING around colonies |
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Beta Hemolysis can be an indicator of a ____________________ .
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Pathogen
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Alpha Hemolysis is indicated by what what 2 things?
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INCOMPLETE lysis of RBC's in blood agar
Greenish/brownish darker color around colonies |
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Gamma hemolysis is indicated by what 2 things?
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NO hemolysis
No change in agar around the colonies |
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Selective media contains___________
that ___________ for the growth of a __________ while ________ the growth of others. |
an agent or media
selects certain group of bacteria inhibiting |
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Selective media can be used to ______ a specific bacteria from a sample containing a __________ .
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isolate
mixed population of bacteria. |
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PEA agar for Gram Positive bacteria and Mannitol salt agar are 2 types of ___________________ media
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Selective
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Differential media in a differential/selective media contains an indicator that is ____________ by __________ of bacteria.
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visibly changed
metabolic activity |
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The more common type of Differential/selective media is ______ and two others that are used are:
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MacConkey's Agar
Eosin Methylene Blue agar (EMB) and Hetoen_Enteric agar |
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MacConkey's agar is commonly found in conjunction with ____________ .
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Blood agar plate (BAP)
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MacConkey's agar is selective for what types of bacteria?
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Gram negative rods (GNRODS), esp. enteric varities such as E. Coli and salmonella
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MacConkey's agar differentiates between ______________ and ___________.
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Lactose fermenters (LF)
Non-lactose fermenters (NLF) |
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GNRODS that ferment lactose are indicated by _______________ on MacConkey's agar.
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Pink colonies
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GNRODS that are non-lactose fermenters are indicated by _________ on MacConkey's agar.
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Colorless
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Eosin Methylene Blue (EMB) agar is similar to MacConkey's and selects for ___________ and differentiates between LF and NLF
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GNRODS
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With EMB, LF are indicated by ________ and NLF are indicated by_________ and E. Coli looks like _______.
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dark, almost black colonies
Colorless colonies colonies that are dark, green with a metallic sheen |
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Bacteria prefer a ___________ ph of ______ with a preferred temperature of ______ for ideal growth conditions but can adapt to extremes.
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basic, slightly alkaline
7 - 7.5 35-37 degree Celcius |
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Aerobes grow _____________ and will be found _______ of a Thioglycollate broth tube.
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in the presence of O2
TOP |
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Anerobes grow ________and will be found growing __________of a Thioglycollate tube.
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in the absence of O2
at the bottom (Tetanus, botulism) |
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Facilitative anerobes have the ability to adapt and change and therefore grow_________ and will be found growing _____________ in a Thioglycollate tube.
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in presence or absence of O2, although better with O2
throughout but more towards the top |
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Microaerophilic anerobes grow in __________ and will be found growing ____________ in a Thioglycollate tube.
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reduced O2 concentration
just below the surface |
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Bacteria are ______________ which means that they are ____________
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ubiquitous
EVERYWHERE |
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Normal Flora are defined as bacteria __________ without causing ________
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living on or in and animal
disease. |
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|
Normal flora are found all over the body but are most prevalent _______________
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in the alimentary tract (mouth, large intestine, rectum)
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|
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Some of the other areas you will find Normal flora bacteria are:
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skin and mucous membranes
Upper Respiratory Tract (nasopharynx) GU tract external regions External eye and external ear canal |
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Mueller-Hinton agar is used for ____________ susceptibility testing.
|
antibiotic
|
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Microaerophilics grow in ______ and in the lab this type of environment is accomplished using a ________
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reduced O2
candle jar |
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You know you have grown a pure culture of only 1 bacteria when you see that ___________ and the type of _________ is the same.
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colony morphology is the same
hemolysis |
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Colony morphology is the ___________ and is important for ___________
|
description of colonies as observed by eyes and not microscopically
ID of bacteria |
|
|
What are 3 types of descriptions commonly used for colony morphology?
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Size-large, med., small, pinpoint
Color-white gray, opaque Consistency- shiny, dull mucoid |
|
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After recording colony morphology, the next step is to do ___________ which is important for beginning to ________________
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A gram stain
ID genus and species as well as to begin antibiotic testing |
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For a mixed culture, the morphology will be ________ and the hemolysis ___________
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different
MAY be different |
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In a mixed culture, the most abundant colony is the _________________ which can be indicated if you see _______ hemolysis.
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pathogen
Beta |
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With a mixed culture, you must initiate a __________
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pure culture
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