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Class:BIOL 201 - Introductory Microbiology - NS, SM
Subject:Biology
University:Pacific Lutheran University
Term:Fall 2009
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Ex.1:Ubiquity of Microorganisms 1. We used trypticase soy agar medi 2. 37=organisms with warm-blooded cells 22-25=organisms that grow on glaciers 3. fungi grows larger and fluffy, while basteria has a soft and glossy appearance and tend to be cream white or yellow colored. >you cannot visualize viruses on an agar plate because there must be bacteria for them to inhit and even there they form plaques.
Ex.2:Pure Culture and Aseptic Technique 1. asceptic technique is when you keep your culture free of contamination from the environment or other organisms. >prevent contamination/infection >easier to observe a certain organism 2.a pure culture is a population of cells resulting from the the growth of a single cell. 3.Because by spreading the bacteria on the surface of a plate the cells are able to form individual colonies separated from other individual colonies. used to obtain a pure culture. 5. No, there is no specific rule based on quadrant.
Ex.3: Introduction to the compound light microscope 2. the field of vision becomes more narrow/decreases as magnification is increased. 4.increase the amount of light when magnification is high and when viewing a stained slide. 5. Iris diaphram, dimer switch, condenser 6. A parfocal is a lense that stays in focus when magnification/focal length is changed. 10. resolution is the ability to distinguish two close objects as distinct from one another rather than as one hazy object. Resolving power is the minimum distance existing b/t 2 objects when those objects
Ex. 5: simple stains-positive and negative stains 1. morphology is the from,structure and configuration of an organism, while arrangement is how the cells are arranged. 2.simple stains is a proceedure for staining bacteria consisting of a single stain while differential stain stains specific morphological structures; usually a multuple stain. 3. a negative stain creates a clear organism on a black background. 4. there are 1000 nanometers in a micrometer and 1000 micrometers in a millimeter.
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Ex. 6: Differential and Special Stains 2. You heat fix a slide to cause the bacteria to adhere to the glass slide. 4. cells are too old or over-discoloration. 5. crystal violet:primary stain (both purple) >iodine solution:mordant (both purple) >alcohol solution: discolorant (gp:puple, gn:discolored) >sufranin solution: counter stain (gp:purple, gn:pink)
Ex. 7: Chemically defined, complex, selective, and differential media 1. Complex/enriched: the exact amount and kinds of large organic molecules are unknown, while chemically defined media is made up of specific amounts of chemicals. Selective media allows only certain bacteria to grow and differential media shows whether a bacteria ferments certain sugars. 3. the organism will not grow well if placed in the wrong media.
What type of media are the following? TSA: complex/rich media (require vitamins & other growth factors) Glucose salts agar: chemically defined media (cell components from glucose and inorganic salts) EMB: selective and differential (gn enteric rods and ferment lactose) McConkey: selective and differential (gn and ferment lactose) Mannitol Salts:selective and differential (ferment mannotol and tolerate high saline concentrations like staphyloccocus) Blood agar: Complex/enriched and differential (alpha and beta blood cells hemolysis)
Ex. 8: Quantification of Microorganisms 2. so plates will have a distinct colonies that can be counted. 3. several dilutions muct be plated and some bacteria grow together creating one colony so aqurate numbers aren't promised, some bacteria have special requirents. 4. No it is not guaranteed to be sterile.
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Ex. 9: Aerobic and anaerobic growth aerobes:organisms that have a requirement for oxygen in order to grow. anaerobes:organisms that will not grow in the presence of oxygen facultative anaerobes: can grow with/without the presence of oxygen. 3. because shake tubes can produce an anaerobic environment, while agar plates only offer an aerobic environment.
Ex. 12: Control of microbial growth using UV light 1. It causes mutations; formation of thymine dimers, 2 adjacent bind to echother causing incorrect base pairing. Ultimately death of a cell if enough radiation is applied. 2. As a control one should use a plate with growth that was not exposed to uv light of the same bacteria. 3. endospores.
Ex. 14: Antiseptics and Antibiotics 1. You measure the clear zone around the antibiotic disk. >measure in millimeters and look at chart.
Ex. 22: Normal skin biota 1. we looked for staphylococcus epidermis and aureus, micrococcus luteus and propionibacterium acnes. >organisms that are found in the skin which help block access to harmful organisms;commensals 2. because staphylococcus are facultative anaerobes. 3. It turns yellow: thos means the organism is able to ferment glucose. 4. staphylococcus and micrococcus will both grow, but only staph will ferment glucose.
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Ex. 16: Transformation 1. the purpose of this lab was to transfer the genes of one bacterium to another. 2. the source of DNA was lysed cells from acinetobacter resistant to streptomycin. 3.because the DNA was not sterile 4. the Str sensative to show that it was sensitive to streptomycin and the Str resistant to show it was resistant the the Str sensative with the DNA proved transformation and DNA showed free of contamintion. all plus DNAse prevent transfor. 5. the quadrant containing Str sensative and the DNA.
Ex. 23: Respitatory Microorganisms 1. We used blood agar because a healthy person will produce a beta hemolysis contaminated plate. 3. A virus won't grow it will take a human cell culture therefore alpha hemolytic streptococcus will grow same as it would on a health person. 4. normal throat flora are alpha and pathogen is beta hemolytic.
Ex. 21:Titering Prokaryotic viruses 2. you will have a completely clear plate because all of the bacteria would have been infected with the viruses. >lawn is the total coverage of bacteria
Ex. 24: Identification of Enteric Gram-Negative rods 1.The tube turns yellow from red. it is necessary to avoid negative data (bacteria could be dead, pH indicator was toxic, didn't metabolize sugar) You must have growth in order to prove the test positive or negative. 2. tells you wheather an organism produces gas or not.
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MRVP Test MR: fermentation and pH VP: presence of acetoin Methyl red: positive is red (pH is lower than 4.0) negative is same color Vogues Proskeur: positive if a brick red precipitate forms MR (+) and VP (-): mixed acid fermentation pathway MR (-) and VP (+): butaneidol fermentation pathway
glucose, lactose, sucrose positive: tube turns yellow from red negative: orange-red gas produced will show on the Durham tube
Citrate Utilization >organism utilizes citrate Positive: tube turns deep blue (pH rises) negative: tube remains green
Motility: can the organism move? positive: turns red wherever it goes negative: red streak only where stabbed
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MacConkey Agar MacConkey (also McConkey) agar is a culture medium designed to grow Gram-negative bacteria and stain them for lactose fermentation. Positive: gram negative frmenting lactose negative: no growth
Urea Hydrolysis >produces enzyme urea positive: turns bright pink negative: yellow
Ocular Lens magnifies the image usually 10-fold
Stage holds specimen
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condenser focuses the light
Iris Diaphram controls the amount of light that enters the objective lense
objective lens a selection of lens options provide different magnifications.
light source controls the amount of light.
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 Ex.1:Ubiquity of Microorganisms1. We used trypticase soy agar medi 2. 37=organisms with warm-blooded cells 22-25=organisms that grow on glaciers 3. fungi grows larger and fluffy, while basteria has a soft and glossy appearance and tend to be cream white or yellow colored. >you cannot visualize viruses on an agar plate because there must be bacteria for them to inhit and even there they form plaques.
 Ex.2:Pure Culture and Aseptic Technique1. asceptic technique is when you keep your culture free of contamination from the environment or other organisms. >prevent contamination/infection >easier to observe a certain organism 2.a pure culture is a population of cells resulting from the the growth of a single cell. 3.Because by spreading the bacteria on the surface of a plate the cells are able to form individual colonies separated from other individual colonies. used to obtain a pure culture. 5. No, there is no specific rule based on quadrant.
 Ex.3: Introduction to the compound light microscope2. the field of vision becomes more narrow/decreases as magnification is increased. 4.increase the amount of light when magnification is high and when viewing a stained slide. 5. Iris diaphram, dimer switch, condenser 6. A parfocal is a lense that stays in focus when magnification/focal length is changed. 10. resolution is the ability to distinguish two close objects as distinct from one another rather than as one hazy object. Resolving power is the minimum distance existing b/t 2 objects when those objects
 Ex. 5: simple stains-positive and negative stains1. morphology is the from,structure and configuration of an organism, while arrangement is how the cells are arranged. 2.simple stains is a proceedure for staining bacteria consisting of a single stain while differential stain stains specific morphological structures; usually a multuple stain. 3. a negative stain creates a clear organism on a black background. 4. there are 1000 nanometers in a micrometer and 1000 micrometers in a millimeter.
 Ex. 6: Differential and Special Stains2. You heat fix a slide to cause the bacteria to adhere to the glass slide. 4. cells are too old or over-discoloration. 5. crystal violet:primary stain (both purple) >iodine solution:mordant (both purple) >alcohol solution: discolorant (gp:puple, gn:discolored) >sufranin solution: counter stain (gp:purple, gn:pink)
 Ex. 7: Chemically defined, complex, selective, and differential media1. Complex/enriched: the exact amount and kinds of large organic molecules are unknown, while chemically defined media is made up of specific amounts of chemicals. Selective media allows only certain bacteria to grow and differential media shows whether a bacteria ferments certain sugars. 3. the organism will not grow well if placed in the wrong media.
 What type of media are the following?TSA: complex/rich media (require vitamins & other growth factors) Glucose salts agar: chemically defined media (cell components from glucose and inorganic salts) EMB: selective and differential (gn enteric rods and ferment lactose) McConkey: selective and differential (gn and ferment lactose) Mannitol Salts:selective and differential (ferment mannotol and tolerate high saline concentrations like staphyloccocus) Blood agar: Complex/enriched and differential (alpha and beta blood cells hemolysis)
 Ex. 8: Quantification of Microorganisms2. so plates will have a distinct colonies that can be counted. 3. several dilutions muct be plated and some bacteria grow together creating one colony so aqurate numbers aren't promised, some bacteria have special requirents. 4. No it is not guaranteed to be sterile.
 Ex. 9: Aerobic and anaerobic growthaerobes:organisms that have a requirement for oxygen in order to grow. anaerobes:organisms that will not grow in the presence of oxygen facultative anaerobes: can grow with/without the presence of oxygen. 3. because shake tubes can produce an anaerobic environment, while agar plates only offer an aerobic environment.
 Ex. 12: Control of microbial growth using UV light1. It causes mutations; formation of thymine dimers, 2 adjacent bind to echother causing incorrect base pairing. Ultimately death of a cell if enough radiation is applied. 2. As a control one should use a plate with growth that was not exposed to uv light of the same bacteria. 3. endospores.
 Ex. 14: Antiseptics and Antibiotics1. You measure the clear zone around the antibiotic disk. >measure in millimeters and look at chart.
 Ex. 22: Normal skin biota1. we looked for staphylococcus epidermis and aureus, micrococcus luteus and propionibacterium acnes. >organisms that are found in the skin which help block access to harmful organisms;commensals 2. because staphylococcus are facultative anaerobes. 3. It turns yellow: thos means the organism is able to ferment glucose. 4. staphylococcus and micrococcus will both grow, but only staph will ferment glucose.
 Ex. 16: Transformation1. the purpose of this lab was to transfer the genes of one bacterium to another. 2. the source of DNA was lysed cells from acinetobacter resistant to streptomycin. 3.because the DNA was not sterile 4. the Str sensative to show that it was sensitive to streptomycin and the Str resistant to show it was resistant the the Str sensative with the DNA proved transformation and DNA showed free of contamintion. all plus DNAse prevent transfor. 5. the quadrant containing Str sensative and the DNA.
 Ex. 23: Respitatory Microorganisms1. We used blood agar because a healthy person will produce a beta hemolysis contaminated plate. 3. A virus won't grow it will take a human cell culture therefore alpha hemolytic streptococcus will grow same as it would on a health person. 4. normal throat flora are alpha and pathogen is beta hemolytic.
 Ex. 21:Titering Prokaryotic viruses2. you will have a completely clear plate because all of the bacteria would have been infected with the viruses. >lawn is the total coverage of bacteria
 Ex. 24: Identification of Enteric Gram-Negative rods1.The tube turns yellow from red. it is necessary to avoid negative data (bacteria could be dead, pH indicator was toxic, didn't metabolize sugar) You must have growth in order to prove the test positive or negative. 2. tells you wheather an organism produces gas or not.
 MRVP Test MR: fermentation and pH VP: presence of acetoinMethyl red: positive is red (pH is lower than 4.0) negative is same color Vogues Proskeur: positive if a brick red precipitate forms MR (+) and VP (-): mixed acid fermentation pathway MR (-) and VP (+): butaneidol fermentation pathway
 glucose, lactose, sucrosepositive: tube turns yellow from red negative: orange-red gas produced will show on the Durham tube
 Citrate Utilization >organism utilizes citratePositive: tube turns deep blue (pH rises) negative: tube remains green
 Motility: can the organism move?positive: turns red wherever it goes negative: red streak only where stabbed
 MacConkey AgarMacConkey (also McConkey) agar is a culture medium designed to grow Gram-negative bacteria and stain them for lactose fermentation. Positive: gram negative frmenting lactose negative: no growth
 Urea Hydrolysis >produces enzyme ureapositive: turns bright pink negative: yellow
 Ocular Lensmagnifies the image usually 10-fold
 Stageholds specimen
 condenserfocuses the light
 Iris Diaphramcontrols the amount of light that enters the objective lense
 objective lensa selection of lens options provide different magnifications.
 light sourcecontrols the amount of light.
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