Micro Lab Practical 1

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duram tube

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open on bottom, closed on top. Tube will turn yellow if sugar is fermented. You will also see a gas bubble because of the CO2 given off

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selective media

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contain ingredients that enhance certain organisms but inhibit others. Ex: methylene blue, crystal violet

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differentil media

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aid in the identification of organisms based upon its appearance of growth.

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nutrient broth(agar)

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general purpose media used to grow bacteria and fungi having basic growth factor needs

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blood agar

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used to detect strep

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Mueller-hinton agar

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recommended for testing antibiotic susceptibility

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Chrom agar

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differential agar that distinguishes urinary tract pathogens based on colony color

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living members of the microbial world

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bacteria, fungi, algea, protozoa and parasitic worms

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non living members

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viruses, viriods, prions

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another term for growing

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culturing

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define media

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food that is used to culture microorganisms

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it is estimated that what percent of microorganisms can be cultivated?

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only 1%

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what are the general requirements for growth?

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water, carbon, hydrogen, oxygen, nitrogen, phosphorus and sulfur

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body temp

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37 C

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room temp

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20-25 C

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refigerator temp

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4-10 C

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freezer temp

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-20 C

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incubator

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chambers used to culture organisms

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isotonic

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concentration of solute is equal inside and outside the cell

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hypertonic

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solute concentration is greater outside the cell than inside the cell. cell shrinks as water rushes out

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hypotonic

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solute concentration I sless outside the cell than inside the cell. cell lysis as water rushes into cell

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solid media

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we use agar instead of gelatin because gelatin will liquefy at 37 C but agar will not.

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liquid media

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also called a broth, has no agar.

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semisolid media

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contains lower percentage of agar than a solid so that it will demonstrate motility

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form of a colony

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or whole colony shape. This is what we see with our naked eyes. Can be round, round with raised margins L form, etc.

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Margin

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the edge of the colony. can be smooth, wavy, swarmy etc

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elevation

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what the colony looks like from the side Ex. flat, raised, etc

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size

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measured in mm.

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texture of colony

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wet, erose, shiny, dull

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pigment

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certain organismsproduce pigments, can be within the colony or diffusing away

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thalum

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what you see as mold

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mold looks

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hairy

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yeasts are

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smooth and colorful

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yeasts cells vs bacterial cells

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yeasts are larger and are shaped like footballs

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SDA

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selective media for molds and yeasts only

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ring

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film of growth clinging to the test tube

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pellicle

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scum like structure that floats on top

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sediment

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organisms may settle and grow at bottom

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flocculent

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visible chunks or flakes

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turbidity

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looks cloudy

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sterilization

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always go proximal to distal

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parfocalism

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if its focused at a lower power, it will be focused a a higher power

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calculating total magnification

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multiply the power of the ocular lens to the objective lens.

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working distance

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distance between the slide and the objective ens.

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field of vision

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what youre seeing in microscope. It goes down as you go to a higher power

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what happens to refraction in oil

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refraction is decreased

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basic dye

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positively charged dyes that are attracted to the negative components of living cells. LIke DNA an RNA

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negatively charged dyes

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cells will look light against a dark background

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simple stain

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uses one positively charged dye to give cells color.

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manevels dye

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green

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differential stain

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uses 2 or more dyes to reveal differences between cells

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coccus

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round, berry shaped

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bacillus

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rod shapped

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spirochetes

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long, very thin, coiled, flexible cells

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spirilla

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large, rigid, curved, wavey or loosely coiled ccells

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vibrio

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look like a comma

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pleomorphism

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organisms that have multiple shapes. Usually don't have a cell wall

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what is a cluster of 4 cells called?

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tetrad

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staphylo

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grapelike cluster

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Step 1. Loop of water, 2. transfer culture to water and smear 3. air dry then heat fix, 4. cover smear with crystal violet then rinse with water. 5. blot dry

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simple stain

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what stain would you use to examine a capsule

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a negative stain. and we do not heat fix

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types of negatively charged dyes

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nigrosin, india ink, eosin, congo red

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step 1. small drop of nigrosin, 2. transfer culture. 3. smear to uniform thickness an air dry

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negative staining

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gram negative cells

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outer lipid membrane is stripped away by the alcohol based decolorizer and so the cell wall will not retain the crystal violet. Look red or pink

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what is the mordant in the gram stain

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iodine.

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what is the counterstain in gram stain

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safranin

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1. heat fix 2. crystal violet 3. rinse with water 4. iodine 5. rinse with water 6. decolorize with alcohol 7. safranin 8. rinse with water 9. blot dry

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gram stain

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sting test method

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if there is stringing it is gram negative.

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vancomycin method

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is there is a zone of inhibition, it is gram positive(vancomycin susceptible) Vancomycin inhibits formation of peptide bonds

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what cells form an endospore

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bacillus and clostridium

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vegetative cell

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metabolically active cell capable of undergoing binary fission

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sporogenesis

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creation of an endospore

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germination

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conversion of an endospore to a vegetative cell

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endospore stain

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you steam these!! endospores(free cells) look green. Vegetiative cells look pink

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1. steam and add malachite green. 2 remove from head and cool for one minute 3. decolorize with water 4 counterstain with safranin 5. rise with water

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endospore stain

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what is the decolorizer in an endospore stain

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water

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manevels stain

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is positive and will stain cells

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congo red stain

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negatve and will stain background. ph below 3.0 looks blue. ph above 5.3 looks red

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example of an indirect stain

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motility medium

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what are direct stains

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flagella, hanging drop, wet mount

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1. Use a tootpick to put a ring of vaseline around the depression 2. transfer culture to a drop of water and cover 3. flip side ovre and lower over sample until vaseline makes contact 4. quickly flip the slide ovre so the drop is suspended in the depression

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hanging drop stain

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TTC

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tetrazolium salt found on motility medium. Turns red in reduced form. Is usually colorless. Serves as a tracking dye

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Quantitaive viable plate counts

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may be used to enumerate the number of living bacteria in a unit volume

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ingrediants in macConkey agar

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bile salts, crystal violet, congo red. Differentiates lactose fermenters from non fermentors. Inhibits gram positive

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Dilution factor

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is always the recipricol of the dilution. w

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color of e coli on chrom agar

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pink

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color of enterococcus faecalis on chrom agar

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blue

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color of proteus mirabelus on chrom agar

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yellow

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Kirby- Bauer

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quantitative analysis of effectiveness of antibiotics.

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Zone of inhibition

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area of no growth. MM. we look at susceptibility to figure out an antibiotics strength