Micro lab

study of patterns of all diseases

A sign is something that people can notice or see, a symptom is something experienced.
An example of a symptom itchiness of a rash
And example of a sign is a red rash that is visible

Direct: The immediate transfer of an agent from a reservoir to a susceptible host by direct contact or droplet spread. Example: sexual contact

Indirect: The transmission of an agent carried from a reservoir to a susceptible host by suspended air particles or by animate (vector) or inanimate (vehicle) intermediaries.
Example: airborne, fomite (like a doorknob)

the first patient found in an epidemiological investigation

To find out the reservoir of the infection so they can find how an infections is being transmitted.

Yes, people may experience the incubation period which lasts about 10 days where they do no experience any signs or symptoms but are still able to spread it because the virus is establishing itself in the new host.

An organism on which a parasite lives.

Condition caused by a microorganism

A physical object that serves to transmit an infectious agent from person to person. Like a doorknob

A sudden widespread outbreak of an infectious disease.

When an infection is spread the person receiving it is a carrier. They also can spread the disease.

Where most people obtain their virus or illness from.

The exist everywhere

37 degrees Celsius

Using a grease pencils or sharpie label the bottoms of your TSA plate with your name, date, plate number, and incubation temperature.

The bottom of the plate/agar side should be facing upwards to prevent condensation

Clump of Baxter growing on solid media. Originated from 1 single bacterial cell

Color, size, optical properties such as shiny, shape (colony form), margin (colony's outer edge characteristics), and elevation (the degree of which the agar is raised)

No, bacteria is apart of you, everyone has a different flora

Injecting a person with a small dose of a virus to help him or her build up defenses to a disease

a petri dish containing a nutrient medium solidified with agar

An organism that feeds on a living host, sometimes harming it.

organism that does not depend on another organism for food or a place to live

Set of practices and procedures that, when followed rigursouly, prevent contamination of our samples with the environment, and prevents contamination of the environment with out samples.

Hospitals, surgery's, research, foo service, dental hygiene, pharmacy, industry

Allows for the growth of large quantities of bacteria

Great for isolating bacterial colonies

Used for the temporary storage or shipment of bacteria.

motility

Used to destroy microbes on heat resistant laboratory tools

used to light bunsen burner

A device used to aseptically transfer and streak microbes in the laboratory. Burn in between use for aespetic technique.

Name of bacteria and your name and degrees

Solid has more visual characteristics and some bacteria grows better in solid environments.

1. Flame the inoculating loop. Hold the culture tube in your non-dominant hand, open it and flame the neck of the test tube as previously described.

2. Insert the inoculating loop into the tube and carefully scrape some of the bacteria growing on the surface of the medium using the loop. Again, ensure that the loop has sufficiently cooled before attempting to remove organisms. Try not to break the surface of the agar during this procedure. You do not need to fill the looped area with culture. A small but isible amount of bacteria in the loop will suffice.

3. Reflame the neck of the tube of the slant culture. Replace the cap and return the tube to the rack.

4. Hold the labelled tube containing the fresh medium and transfer the bacteria into the new tube of media as follows:. A. If you are inoculating the broth, dip the loop into the broth and gently move the loop back and forth. B. If you are inoculating an agar slant, gently streak the loop along the surface of the slant.

5. Completely flame the loop in the Bunsen bur to Fresh Media

1. Examine the agar plate carefully before you begin. The solid media is contained in the bottom portion of the plate.
2. Flame the inoculating loop as described previously.
3. Keep the plate on the table with the lid up. Without moving the plate, Iift the lid of the Petri dish so that it makes a 45° angle with the bottom of the plate. Do not remove the lid entirely, because this makes contamination of the agar much more likely.
4. Insert the loop between the lid and the bottom of the plate and gently scrape a small amount of growth from the agar without gouging the agar in the process. You do not need to fill the loop with the culture; remember thata tiny amount will have millions of organisms.
5. Replace the lid on the plate and hold the labelled tube containing the fresh medium. Open the tube and flame the neck. A. If you are inoculating the broth, dip the loop into the broth and gently move the loop back and forth. B. If you are inoculating an agar slant, gently streak the loop along the surface of the slant and transfer the inoculum to fresh medium.
6. Completely flame the loop in the Bunsen burner.

Microorganisms that should not be present within, or on the sample or item that you are working with in the lab.

Eliminating all living biological contaminates

Same and incoculating loop used to aesoeticly transfer bacteria

The solid or liquid substratum on which, or in which, cells or organ explants can be made to grow agar or broth

one that contains more than one type of organism growing in a sterile medium, such as agar. The mixed culture can include multiple species of viruses, bacteria and parasites, which may or may not live in harmony with one another, sharing the available resources.

laboratory culture containing a single species of organism.

the cloudiness or haziness of a fluid, originated by suspended insoluble particles.

Improve contrast to see bacteria better

Does not identify the classification of bacteria. it only gives limited information about the morphological characteristics.

Over mixing bacteria in water
Amount of bacteria
Allows for better staining

Adheres bacteria to slide
Allows for better staining

Simple: simple dye, only for contrast, color does not matter

Differential: 2 or more dyes, observe specific characteristics, color does matter, gram positive-purple, gram negative-pink.

spherical

Diplococcus: paired
Streptococcus: chains
Tetrads: four sided
Staphylococcus: grape like clusters
Sacinae: cubical packets

rod shaped

Arrangements:
Dipolesbacillus: paired
Strep to bacillus: chains
Palisading: side by side
Vibrio: 1/2 spiral turn

a staining dye/indicator that interacts with nucleic acid molecules and proteins, turning them to a very dark blue color

counter stain in gram stain

one hand on the neck and one hand on the base

spiral

Spirillum: loose spiral
Spirochete: spiral

difference between light and dark

Adjust light intensity and stain

Magnification makes it larger so you can see whereas resolution provides clarity

ocular x objective

Turn off the microscope lamp, and HOLD the plug while removing it from the outlet. DO NOT PULL ITOUT BY THE CORD.
2. Rotate the nosepiece until the scanning objective (4X) lens is in place over the stage.
3. Using the course adjustment knob, lower the stage to the lowest possible height.
4. NEVER leave your microscope with a slide on its stage. If you were using a prepared slide, wipe off any fingerprints or oil that is on the slide with lens paper before you return it to its storage cabinet. Dispose of your stained slides in the appropriate container for used slides.
5. Wrap the power cord around the microscope as shown by your instructor.
6. Clean the microscope assigned to you. Use lens paper and lens cleaner to clean the mechanical stage, all of the objective lenses, the ocular lenses, and the condenser lens. Make it a habit to clean your oil immersion lens last so that you do not accidentally transfer any oil from the 100X objective to the other objective lenses. A. Fold your lens paper up to make a postage-stamp-size square and dab it against the end of the lens repeatedly. Fold the paper over to expose a fresh portion and repeat the dabbing. B. If there is a significant amount of oil still trapped in the lens or if there is oil in/on any of the other objective lenses, apply a lens cleaning solution to the lens paper and wipe again. Dry the lens with 95% ethanol since this will dry easily.
7. Cover the microscope, and return the microscope to its storage cabinet.

Increase light intensity by opening the Otis diaphragm or turning up the Eurostar as the magnification increases

Use fine focus if necessary to focus

Clean lens

Slide might have not been focused properly on the scanning power lense so start over.

a glass slide holding a specimen suspended in a drop of liquid (as water) for microscopic examination

The area visible through the microscope eyepiece

Property of microscope which allows objectives to be changed without having to refocus

a knob that makes large adjustments to the focus.

100x objective lens

provides a magnification of 40x and is the longest objective

provides a magnification of 10x and is the shortest objective

4x magnification

scanning (4x), low power (10x), high power (40x), oil immersion (100x)

Holds objective lenses and rotates

Eyepiece of a microscope

controls the amount of light passing through the specimen

microscope component that focuses light on objects under study

supplies the light required to view the specimen

Holds the microscope slide in position.

microscope that allows light to pass through a specimen and uses two lenses to form an image

used to distinguished between different types, or groups, of bacteria

Gram Positive bacteria (will look purple because they retain the primary stain after the decolorizer)

Gram Negative bacteria (will look pink from safranin because they were decolorized)

Since all bacteria are either gram +/- this stain is the first thing used to determine what type of bacteria is present in the specimen.

This helps us figure out what organism we are dealing w/. the results are recorded as Gram +/-

Cell wall

Gram positive- this cell wall peptidoglycan (50%)

Gram negative has a thin Cell wall COMPOSED OF ONLY 15% if peptidoglycan and is composed of lipids, lipopolysaccharides.

diplobacilli and pink

staphylococcus and purple

crystal violet; Initially, both gram + and gram - cells are stained by this to retain the primary stain

forma a complex with a crystal violet and traps it in the gram positive

destain only the gram negaitve

to see the gram negative so provides an alternative to see because otherwise it would be clear.

All bacteria will appear gram negative because color such as purple will be very light and appear pink

all bacteria will appear Gram positive (not enough to remove the outer membrane of Gram negatives).

Expired culture smears: If culture smears are too old, the bacterial cell wall might have already broken down, rendering them incapable of retaining the Crystal Violet stain. All bacteria would, therefore, appear Gram-negative.

you will wash off your bacteria, leaving little to none to be seen under the microscope.

Evades phagocytes, prvents phocytosis, helps with attachment. observation of capsules in bacteria. On the other hand, with the help of flagella staining, one can detect the presence of flagella in a bacteria

allows bacteria to evade phagocytosis which makes it harder for the immune system to attack bacteria causing an infection

Bacteria: Mycobacterium tuberculosis

The peptidoglycan prevents osmotic lysis. Layer 2: The arabinogalactan layer is linked to both the peptidoglycan and to the mycolic acid outer membrane and probably provides additional strength to the cell wall.

helps the bacteria survive in harsh enviorments or conditions

surrounded by an outer covering which is proteinaceous. This coating provides enzymatic and chemical resistance to the spore.

Bacterial flagella structure: very thin and hard to see under microscope

function: motility and attachment

satin reddish/pink, while non-acid fast bacteria stain blue

Endospore stain green, while vegetative cell stain reddish/pink

Capsules are colorless in structures found on the outside of the stained bacteria.

uses mordant thar causes flagella to swell and become incrusted with stain allowing it to be easily seen.

By using different stains, one can preferentially stain certain cell components, such as a nucleus or a cell wall, or the entire cell.

Acid fast: heat bacteria in the primary stain

2) different dyes

3) different decolorizer- acid alcohol

Mycobacterium

The structure of interest remains unstained, while the rest of the cell background are stained.

it enhances the ability of bacteria to cause disease

Monotrichous (one tail)

Amphitrichous (two tail)

Lopotrichous: MOre than one tail from one side)

Petritrichous: Multiple all over

No, because it is not stressed yet so it still has nutrients to survive.

acid fast

waxy substance found in the cell wall of mycobacterium

acid-fast or non acid-fast will be Fuchsia

Red dye containing phenol.

primary stain in acid fast stain

staining the background instead of the cell

gram positive rods (bacilli)
-anaerobe

primary stain in endospore stain

technique used to separate one species from another based on morphological differences

1 bacterial cell

Pure: 1 type of microbe

Mixed: 2 or more types of microbes

Pure

To isolate bacteria and identify unknown bacteria from a mixed culture

When wanting to know the approximate numbers of bacteria in a give sample

Pour plate is quantitative so you are able to find out the number of bacteria

To kill or minimize the amount of bacteria vein's transferred from quadrant to quadrant

Reduce bacteria to get an accurate count. Most common # of colonies: 30-300

Colony forming units estimate of the number of bacteria that can form a colony which mean it's is living cause these are the ones that are causing harm

To count the number of bacteria that form a colony because this means they are alive and toxic to humans

number of colonies that develop provide estimate of the total population

Agar plate because it has a large surface area