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BSL-1 Laboratory Practices
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Standard microbiological practices are followed.
Work can be performed on an open lab bench or table. (Bacillus subtilis, E. coli, rhodospirillum rubrum, Lactobacillus acidophilus)
Work can be performed on an open lab bench or table. (Bacillus subtilis, E. coli, rhodospirillum rubrum, Lactobacillus acidophilus)
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BSL-2 lab practices
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organisms are commonly founding the community. Ingestion routes are ingestion, inhalation, and penetration of skin. (salmonella, staphylococcus aureus, clostridium dificile, borrelia burgdorferi)
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BSL-3 lab practices
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Organisms are local or exotic associated with respiratory transmission and lethal diseases. Only specifically trained personnel can handle microbes and access to lab is restricted. (bacillus anthracis, mycobacterium tuberculosis, West Nile virus)
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BSL-4
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Microorganisms that are dangerous and exotic with high risk of aerosol transmitted infections. Rarely are there treatments or vaccines for these microorganisms, and the diseases they cause are frequently fatal. (Ebola virus, Marburg virus, Lassa fever)
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What kind of lab does micro 205 have?
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BSL-2
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Controlled experiment
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all variables except one, experimental variable, are maintained without change
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cause-and-effect relationship
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changes in one variable cause the changes in the other variable; can be determined only through an experimental research design
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Where can microorganisms be found?
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soil, water, plants, animals, humans, some uninhabitable places
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pathogens
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causes damage to host
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mutualism
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both host and microbe benefit
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Commensalism
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microbe benefits but has no affect on the host
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opportunistic pathogens
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do not cause disease under normal conditions but cause disease under special conditions (immunocompromised)
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What is the purpose of blood agar?
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used to differentiate bacteria based on their hemolytic characteristics
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beta hemolysis
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complete lysis of red blood cells
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alpha hemolysis
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partial hemolysis
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gamma hemolysis
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no hemolysis
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Transfer broth culture to sterile broth
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1. label sterile broth
2. flame/ incinerate loop
3. pick up culture and flame edge of tube
4. with loop grab a sample (flame culture tube before placing back on rack)
5. pick up sterile tube and flame edge
6. place loop in the broth to mix organism and sterile broth
7. flame the edge of mixed culture
8. flame loop
2. flame/ incinerate loop
3. pick up culture and flame edge of tube
4. with loop grab a sample (flame culture tube before placing back on rack)
5. pick up sterile tube and flame edge
6. place loop in the broth to mix organism and sterile broth
7. flame the edge of mixed culture
8. flame loop
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Transfer from agar slant to sterile agar slant
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1. label sterile agar
2. flame loop
3. pick up culture and flame the edge of tube
4. touch sterile loop to agar surface to pick up growth (flame culture tube before placing back)
5. pick up sterile agar slant and flame edge
6. touch agar with loop and move loop in a zigzag pattern of surface
7. flame the tubes edge and return to rack
8. flame loop
2. flame loop
3. pick up culture and flame the edge of tube
4. touch sterile loop to agar surface to pick up growth (flame culture tube before placing back)
5. pick up sterile agar slant and flame edge
6. touch agar with loop and move loop in a zigzag pattern of surface
7. flame the tubes edge and return to rack
8. flame loop
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transfer from plate culture to sterile broth
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1. label sterile tube
2. flame loop
3. touch loop to culture agar
4. pick up sterile tube and flame edge
5. place loop in sterile broth (flame edge after)
6. flame loop
2. flame loop
3. touch loop to culture agar
4. pick up sterile tube and flame edge
5. place loop in sterile broth (flame edge after)
6. flame loop
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Quadrant streak plate
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what is the name of a method used as a means of isolating organisms from a mixed culture, pulling organism from each section to end in isolation
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spread plate method
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a plate count method in which inoculum is spread over the surface of a solid culture medium
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flocculent growth in broth
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suspended chunks or pieces
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sediment growth in broth
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growth on the bottom
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ring growth in broth
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growth at the top around the edge
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Pellicle growth in broth
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membrane at the top
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Uniform fine turbidity (broth)
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evenly cloudy throughout
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filiform growth
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smooth texture with solid edge
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spreading edge
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solid growth seeming to radiate outward
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transparent growth
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almost invisible
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friable growth
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rough texture with crusty appearance
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pigmented growth
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produces colored growth
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decontamination
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reduce pathogens to levels considered safe to handle, first step towards disinfection, includes physical cleaning with soaps or detergents
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Disinfection
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A process that eliminates many or all microorganisms, with the exception of bacteria spores, from inanimate objects, typically a chemical agent but can be dry heat, moist heat, UV light
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antiseptic
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disinfectants that are designed to reduce or eliminate pathogens on or in living tissue
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Sterilization
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the complete elimination or destruction of all microorganisms, including spores, the highest level of pathogens control
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P- 200 pipette
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20-200 ul
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P-1000 micropipette
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100-1000 microliters, blue tips
first column(red) - thousands
second column - hundreds
third column - tens
first column(red) - thousands
second column - hundreds
third column - tens
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Autoclave tape indicator
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Used to show that the instruments have been properly sterilized by changing colors. and black stripes appear
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total magnification
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objective lens x ocular lens
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cocci
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spherical bacteria
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bacilli
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Rod shaped bacteria
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spirillum
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spiral shaped bacteria
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tetrad
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division of cocci into a square shape
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simple stain
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use of a single basic dye, helps see morphology
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gram stain
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A staining method that distinguishes between two different kinds of bacterial cell walls.
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order of stains in the gram stain procedure
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1. crystal violet (rinse)
2. iodine (rinse)
3. decolorizer/ alcohol
4. safranin
2. iodine (rinse)
3. decolorizer/ alcohol
4. safranin
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crystal violet stain
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Primary stain, gets stuck in the cell wall (peptidoglycan)
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iodine stain
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mordant, to enhance crystal violet staining
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decolorizer in gram stain
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Alcohol, removes primary dye from Gram-negative cell
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Safranin stain
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Added as the counter/secondary stain in a Gram Stain which color the gram negative bacteria pink-red and have no effect on gram positive bacteria.
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Over decolorized gram stain
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get reddish gram positive cells
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Underdecolorized gram stain
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variability in the stain overall
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negative stain
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colors the background, which makes capsules more visible using nigrosin, used to view bacteria that cannot handle heat
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antimicrobials
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used to treat infectious diseases
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antibiotics
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natural antimicrobial agents produced by microorganisms (penicillin)
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Bacteriocidal
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agent that kills bacteria
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Bacteriostatic
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inhibits bacterial growth
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Kirby-Bauer Test
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The test to determine the efficacy of antibiotics or the antibiotic resistance of bacteria. The bigger the zone of inhibition, the more effective
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blood typing
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The blood of one person is different from another's due to the presence of antigens on the surface of the erythrocytes. The major method of typing blood is the ABO system and includes types A, B, O, and AB. The other major method of typing blood is the Rh factor, consisting of the two types, Rh+ and Rh-.
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universal donor
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O negative
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universal recipient
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AB positive
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pour plate
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Diluted sample mixed with agar, poured into empty petri dishes. Colonies form in and on plate.
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transfusion reaction
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a serious, and potentially fatal, complication of a blood transfusion in which a severe immune response occurs because the patient's blood and the donated blood do not match
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