Lab Midterm Test Questions – Flashcards
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Unlock answersFomite |
Any object, surface, or body part |
Reservoir |
Any area where a microbe resides and serves as a potential source of infection |
Resolution |
The ability to distinguish two adjacent points as separate objects based upon
1. Numerical aperture 3. Design of condenser |
Numerical aperture (N A) |
the measure of a lens’ ability to capture light coming from a specimen and use it to make the image; increases with the use of oil and the higher objectives |
Limit of Resolution |
(N A condenser) + (N A of objective lens) |
Phase Contrast vs. Brightfield Microscopy |
- usually organelles and cells are transparent and difficult to differentiate without staining - staining kills the cells so you don’t see living cell just artifacts - phase-contrast allows one to view living transparent organisms using phase (wavelength shifts) principles of light to produce dark objects in a bright background. |
Cyanobacteria |
- blue-green organisms - belong to Kingdom Monera - prokaryotic type of nucleus - Cyanobacteria: chlorophyll a - phototrophic bacteria:bacteriochlorphyll - Cellular structure is considerably different from the eukaryotic algae |
Protozoa |
- unpigmented, motile organisms - belongs to Kingdom Protista - eukaryotic - single-cell transparent organisms, with a cell membrane, and no cell wall - in lab, paramecium are the most common to be seen |
Algae |
- greenish or golden brown organisms - belongs to Kingdom Protista and includes all the photosynthetic eukaryotic organisms - no tissue differentiation - distinct, visible nuclei and chloroplasts - unicellular or colonial |
Purpose of Heat-fixing |
1. To kill the bacteria 2. To fix them to the slide 3. To coagulate cytoplasmic proteins to make cells more visible |
Basic dyes |
- have positive (+) charge - ex. Methylene blue - can penetrate the bacteria because of the attraction between charges |
Acidic dyes (negative stain) |
- have negative (-) charge -chromophores have negative charge |
Capsules |
Function
|
Spore Staining: Schaeffer-Fulton Method |
1. Make a smear of each bacteria: B. stearothermophilus, C. sporogenes, and S. epidermidis (either on separate slides or 1-2 slides). 2. Setup a Bunsen burner on your ring stand with the ring clamp and wire gauze over it. 3. Fill the can (from your student drawer) with water from the tap until it is about half-full. Place the can on top of the wire gauze on the ring clamp. 4. Turn on the Bunsen burner to heat/boil the water. (Let steam start to appear before putting the slide on the can.) 5. Place the slide with smear(s) on it on top of the can 6. Place a piece of paper towel on top of the smear on the slide and saturate the paper with Malachite green. 7. Steam the smear over the boiling water for 5 minutes. NOTE: If the Malachite green evaporates off, then add more - Keep the paper wet. Use forceps to take the slide off the can when finished - it will be hot!! 8. Allow the slide to cool, remove the paper, and rinse with water for 30 seconds. 9. Add Safranin to the slide for 1 minute. 10. Rinse off the Safranin with water. 11. Blot the slide between sheets of Bibulous paper. 12. Examine the slide under oil immersion. |
Acid-Fast stain |
Differential staining technique that it used to detect the presence
Mycobacterium |
Gram Stain |
Components: Crystal Violet - Primary stain - stains all the bacteria initially Gram’s iodine - mordant - forms an insoluble compound upon interacting with crystal violet 95% Ethanol - decolorizing agent - washes away the dye in Gram-negative bacteria and does not touch the dye in Gram-positive bacteria Safranin - counterstain - stains all the bacteria - Gram-negative bacteria appear red under the microscope |
Amphitrichous |
ex. Spirillum volutans (flagella at both ends) |
Monotrichous |
ex. Pseudomonas fluorescens (a single flagella at one end) |
Brownian motion |
organism vibrates due to molecules hitting the cell, i.e. H2O, etc.; organism will remain in one place |
true motility |
directional; independent movement over greater distances |
chemotaxis |
movement towards chemical attractants (ex. food) and away from repellants (ex. chemicals) |
Motility Test Medium contains TTC (2,3,5-triphenyl tetrazolium chloride) |
TTC is:
|
Advantage / Disadvantage of Motility Test Media |
Advantage of Motility Test Media:
|
A good solidifying agent |
1. Is not utilized by the microorganisms 2. Doesn’t inhibit bacterial growth 3. Doesn’t liquefy at room temperature |
Nutrients |
chemical compounds that can be broken down into or used to make monomers |
Synthetic (Defined) Media vs. Non-Synthetic (Complex Media) |
Synthetic or defined media – exact composition is known; has been synthesized; chemical
Non-synthetic or complex media – the exact composition is not known; utilizes compounds |
Minimum requirements for Steam Sterilization |
For 1 Liter of media: 15 minutes at 121.6oC or 250oF and 15psi |
Selective media |
media that allows only certain organisms to grow
a. Absence of certain nutrients which prevent the growth of some but not all bacteria |
Differential media |
media that contain substances which change the appearance of one species allowing for differentiation ex. EMB – Eosin Methylene Blue - both selective and differential - methylene blue inhibits Gram (+) bacteria thereby selecting for Gram (-) bacteria - eosin- an acidic dye which causes coliforms (bacteria found in the intestine) to turn a metallic green color |
Minimal media |
media that contains the basic requirements for the bacteria to grow such as salts, a carbon source, and a nitrogen source |
Enriched media |
media that contains all the basic requirements but has added amino acids, vitamins, and minerals from such sources as peptone or yeast extract |
Methods of Sterilization |
Autoclaving - Steam Sterilization; utilizes high temperature and pressure UV Irradiation - utilizes UltraViolet light Ethylene Oxide Gas - generally used for plastic surgical supplies and disposable medical supplies that can not be autoclaved |
Streak plate – four streak techniques |
a. Quadrant streak – we use a modified form of this method b. Radiant streak c. ‘T’ streak d. Continuous streak |
Pour plate |
use plates and agar tubes to isolate loopfuls of bacteria; requires less skill and practice but a lot of media and supplies |
Phenyl Ethyl Alcohol (PEA) Agar |
- selective media - Selects for Gram positive bacteria - inhibits the growth of Gram negative bacteria because the phenyl ethyl alcohol hinders their DNA synthesis - used to grow Gram positive anaerobes if 5% Sheep blood is added |
Mannitol Salt Agar (MSA) |
- selective and differential media - contains mannitol, 7.5% NaCl, and the pH indicator Phenol red |
MacConkey Agar |
- selective and differential media - select and differentiate members of Enterobacteriaceae - contains lactose, bile salts, neutral red, and crystal violet |
Eosin Methylene Blue (EMB) |
- is a selective and differential media - contains peptone, lactose, sucrose, eosin, and methylene blue |
Hektoen Enteric (HE) agar |
- is a selective and differential media - isolate Salmonella and Shigella species from other enterics
- contains peptone, lactose, sucrose, salicin, bile salts, sodium thiosulfate, ferric ammonium |
Mycology is the study of |
fungi |
Characteristics of Fungi |
Eukaryotic 2. Non-motile 3. Non-photosynthetic 4. Lack tissue differentiation 5. Have cell walls of chitin or other polysaccharides; not of peptidoglycan 6. Reproduce through spores which are for reproductive purposes only and do not give the fungi resistance to anything like the endospore does. 7. Saprophytic – decompose dead organic matter 8. Heterotrophs – produce exoenzymes that digest nutrients in the environment then absorbs the products |
Molds |
mycelium - mass of intermeshed hyphae; what is seen macroscopically - hyphae - microscopic filaments that may (septate) or may not (nonseptate) have walls - can form either asexual or sexual spores or both |
Yeasts |
have pseudohyphae which are multicellular structures produced through budding - spores - produced by budding which is an outpouching of a cell - smaller in size than the parent cell - may or may not stay attached to the parent cell |
Sabouraud’s Dextrose Agar (SDA) |
- grows molds - has a pH of 5.6 which inhibits |
Slide-mold culture |
method is used because wet mounts and simple transfer of hyphae does not provide the structures necessary for identification due to distortion or destruction of these structures during transfer. This also allows conidiophores to grow on a single horizontal plane |
Plate Count - Direct Measurement |
Dilute the organisms and count them on microscopic fields on a slide. |
Plate Count - Indirect Measurement |
1. Quantitative Plating (Standard/ Viable Plate Count)
|
Quantitative Plating (Serial Dilutions) |
info on the viable organisms is obtained - easy to perform but uses lots of glassware and media and takes time to get results - can calculate the number of organisms in a bacterial culture - must use aseptic technique when handling the pipettes |
Dilution Factor |
|
Final Dilution Factor |
= product of individual dilutions |
Original Cell Concentration |
(volume plated) (FDF) |
CFU |
Colony Forming Units (or the number of viable cells that can make a colony) |
Turbidity Measurement |
(uses a spectrophotometer) -information on both living and dead cells is obtained -easy to perform and uses less glassware and media |
Serological pipette |
pipettes that have graduations down to the tip.
|
Mohr pipette |
pipettes that are not graduated down to the tip so fluid must only be expelled to the last calibration line |
Viruses vs. Bacteria |
- viruses are smaller, non-cellular, and intracellular parasites - viruses can not be grown on normal media because they require a host since they have their own DNA or RNA but lack the cellular machinery needed to replicate - viruses can infect all types of cells: prokaryotic and eukaryotic |
Phages |
viruses that infect bacteria; also called bacteriophages Phages that parasitize E. coli are referred to as coliphages. |
Lytic viruses |
considered virulent; viruses enter the cell, take over the cell, replicate, then lyse the cell for release |
Lysogenic viruses |
considered temperate; viruses enter the cell, integrate into the DNA of the host; cell replicates normally then eventually viruses replicate and lyse the cell to be released |
VIU |
Virus Infection Units Smallest unit causing an effect with a host |
1 plaque = |
1 virus |
PFU |
- Plaque Forming Units - Efficiency of count from the assay is always less then with an electron microscope because efficiency of infection by viruses is not 100% |