Lab 1 Flashcards
Biosafety levels
- no known risk - open bench, sink goggles, gloves
- disease in healthy adults but easily contained, transmission through ingestion, mucous membrane - biohazard warning, sharps, disposal of waste
- Disease may be lethal - controlled access
- high risk of lethal - complete isolation
nosocomial infection
infections acquired while receiving health care that was not previously there
Transient flora
Resident flora
Transient flora - temporarily present - easily acquired and transferred in a healthcare setting,
Resident flora - is always present - in skin and hair follicles, protect us from pathogens, and is difficult to remove. Compete for nutrients, occupy space and produce antimicrobial compounds
Symbiotic -
Commensals -
Opportunists -
Symbiotic - Living together
Commensals - inhabit bodies and may benefit normal flora and opportunities
Opportunists - capable of causing disease when given the opportunity
Culture medium?
Broth?
Solid/plate?
Growth?
Culture medium - nutrient source to allow microbes to grow
Broth - liquid at room temp; growth turbidity
Solid/plate - broth + agar (causes media to become solid)
Growth - colonies - visible masses of microbes
Types of media
General-purpose media?
Enrichment media?
Selective media?
Differential media?
incubation?
General-purpose media - grows most types of microbes that do not need special growth factors *Tryptic soy broth and agar
- *Enrichment media** - special growth factors required by some microbes
- *Blood agar blood cells are broken to get nutrients like heme*
- *Selective media** -substances that inhibit some growth and encourage others
- *Sabourauds dextrose agar - encourages yeast and mold*
- *Differential media** - id of microbe based on physical growth differences (what they look like when growing)
- *Blood agar as different patterns of hemolysis can be observed*
TSA - Tryptic soy Agar
SDA - Sabourauds dextrose Agar
BAP - Blood Agar
TSA - Tryptic soy Agar - general purpose
SDA - Sabourauds dextrose Agar - selective media, inhibits the growth of bacteria due to high concentration of dextrose, promotes the growth of fungi
BAP - Blood Agar - enrichment and differential media, contains growth factors for certain microbes and shows differences between different types of microbes
3 types of Microscopy
Light - magnify images of microorganisms
Brightfield - transmits light through the specimen and appears dark on a light background. Used for stained specimens
Phase-contrast - special lens, diaphragms to slightly alter paths of light waves from an object using diffraction pattern that allows viewing of transparent parts. Used for live, unstained specimens
What are the magnifications of the following
Ocular lens?
Objective lens
Scanner?
Low power?
High dry?
Oil immersion?
Condenser lens?
Ocular lens - 15X
Objective lens
Scanner- 4x
Low power - 10x
High dry - 40X
Oil immersion-100x
Condenser lens - no magnification
What is the total magnification for the following
Ocular + scanner?
Ocular + low power?
Ocular + high dry?
Ocular + oil immersion?
Ocular + scanner = 60X
Ocular + low power = 150X
Ocular + high dry = 600X
Ocular + oil immersion = 1500X
Resolution?
Max resolution of light microscope?
Resolution - ability to distinguish objects from one another
Max = 0.2 micrometers
*particle of light must pass between 2 objects.
*a particle of light is 0.2
Magnification?
How does it work?
What cannot be seen with a light microscope?
Magnification - the ability of a microscope to make an object appear bigger
light passed through specimen picking up image - which lens makes bigger.
if under 0.2 light will just go around it
Viruses cannot be seen with light microscope must use Electron microscope
Parfocal?
Working distance?
Parfocal - the image will stay in focus if you go from lower to higher power objective lens
Working distance - the distance between the bottom of the objective lens and the slide
Prokaryotic cells
Classification?
2 types?
Small, single-celled (unicellular)
Lack true nucleus and membrane-bound organelles
Bacteria and Archaea
Eukaryotic cells
Classification?
Larger, single-celled or multicellular
Nucleus and membrane-bound organelles
What are 3 domains?
What are 4 kingdoms?
Domains - Archaea, bacteria, eukarya
Kingdoms - Protista, Fungi, Animalia, Plantae
Kingdom Protista includes?
Protista
Protozoans and algae
Kingdom Fungi includes?
Dimorphic?
Fungi
Mold - multicellular, filamentous, sexually via spores
Yeast - unicellular, round cells, asexually via budding
Dimorphic - occurs in both forms depending on environment
Kingdom Animalia includes?
Helminths - parasitic worms
What is the procedure for culture spill?
- Cover with paper towel, soak with disinfectant, let stand for 10 mins
- reclean with new paper towel and dispose of all paper towels in autoclave bag
*if available, activated peroxide wipes
What is procedure for fire?
In case of fire, students should inform the instructor immediately, and evacuate the area. Pull the fire
alarm. Inform the CSN campus security officer on duty.
ubiquitous -
Fomite -
can be found in any habitat on earth
fomite – a person or object
that mechanically carries microbes from one person or place to another
What is a colony on an agar plate?
Larger hairy - Mold
Smaller - bacteria
Resident flora must have these conditions to live
Slightly hypertonic
Slightly acidic 5-6
Tolerate oils
tolerate keratinocytes that have little nutrients
Where is the following found:
Staphylococcus aureus
Staphylococcus epidermidis
Corynebacterium
Propionobacterium
Staphylococcus aureus - found on nares
Staphylococcus epidermidis - found on skil
Corynebacterium - on the skin
Propionobacterium - in sebaceous glands
Nosocomial infections may include:
What 3 others can lead to food poisoning?
MRSA - methicillin-resistant S aureus
VRE - Vancomycin-resistant Enterococcus
Clostridium difficile
Can lead to food poisoning:
Staphylococcus
E. coli
Salmonella
Describe the proper way to wear a lab coat?
Where are lab coats stored?
Long sleeves and come to knee level. button up
Stored in the lab with names on them. Stored in bin
What is biosafety level 1 & 2
Level 1 is not known to cause disease in healthy humans, open bench, sink, goggles, gloves
Level 2 is associated with human disease in healthy humans. Easily contained. Transmission through percutaneous, ingestion, mucous membrane.
-Biohazard warning, sharps, disposal of waste and surveillance, biosafety cabinet for aerosols or splash. Googles, lab coats, gloves. plus autoclave
When should Goggles be worn?
Googles should be worn when working with liquids
What is the procedure for a culture spill?
Cover spill with a paper towel
Soak with disinfectant immediately
let stand for 10 mins, reclean with a new paper towel
Dispose of paper towel in an autoclave bag
What is the procedure for a fire?
inform instructor
evacuate area
Pull fire alarm
Inform CSN campus security
What is the last thing a student must do before leaving the lab?
Wash hands
Who was the first person to link the infectious disease, childbed fever, with microorganisms spread through contact with contaminated and unwashed hands
Ignaz Semmelweis
What does cultural media provide organisms?
What makes a solid media solid? what are the advantages of solid media?
What Is a colony on a agar plate?
Nutrient source
Solid media contains agar (dervied from seaweed) & advantages are it provides a flat solid surface
Colony - clones from one bacterium
Should one be concerned about bacteria growth in the throat?
Some can be in the throat temporarily like Streptococcus pyogenes which causes strep throat
What is the difference between a bacterial colony and a microbial colony?
Larger hairy colonies are MOLD
Smaller non fluffy colonies are BACTERIA
What is the purpose of immersion oil?
Reduces bending and distortion.
Oil has the same refractive index as glass so less light bends preserving good resolution
What are the 3 lenses on the microscope and what is the purpose of 3rd lens?
What is the purpose of the iris diaphragm?
Ocular, objective, and condenser. Condenser is not involved in magnification but focuses and concentrates the light reaching the specimen.
The iris diaphragm is within the condenser and can be opened and closed with a lever to regulate the amount of light reaching the slide
What is resolving power?
0.2 micrometers are also called
is the minimal distance 2 objects must be apart in order to distinguish them as separate and distinct
microns
Archaea and bacteria are generally ____-Cellular?
Protista has both ___-cellular and ____- cellular?
Fungi have ____ which are unicellular and ____ which are multicellular
Kindgom Animalia are ___ cellular
Protozoa are classified by
Archea & bacteria - unicellular
Protista have both unicellular and multicellular
Fungi - yeast - unicellular, mold - multicellular
Animals are multicellular
Protozoa are classified by their method of locomotion (Cilia, pseudopodia, flagella, non-motile)
What are the 2 members of the animal kingdom which are eukaryotic cells…
Turbatrix aceti (vinegar eels) that grow well in acetic acid and are motile
Planaria which motile free-living flat worms with eyespots
Fungi are non-photosynthetic _____trophic
Yeasts are best known for their role in
When hyphae become large macroscopic mass they are called
Heterotrophic (use organics for carbon and energy) decomposers
Fermentation in making bread and beer
Mycelium
