Exercise: Media Types (See Handout, not in book)



Exercise 11: The Importance of Handwashing

The goal of this experiment is to:

Identify the types of flora found on different layers of skin.

To compare methods of cleaning hands.

1. Labeling agar plates

We label plates with ______________, _____________,

_________________________, and

__________________________________.

We label plates ______________________________ so if lids are mixed up or turned, we still know what we have.

We label plates _____________________________ so the view is not blocked.

2. We store plates __________________________________, so that water from condensation during the incubation process will collect in the lid, not on the culture. It can be very messy, and can contaminate other samples.

3. _______________________ is the most effective way to prevent the spread of disease!

Review Ex 11: Handwashing

1. _______________________ microorganisms are transient. These pathogens can be transmitted from medical provider’s hand and equipment to patients.

2. ________________________ infections are infections that are acquired in a healthcare setting.

3. ________________________ are microorganisms that are found deep in skin.

There are 4 groups of normal flora:

1. ________________________________

2. ________________________________

3. ________________________________

4. ________________________________

5. Normal flora is beneficial to the host (us) because they protect us from pathogens by:

1.

2.

3.

6. See handout on Shigellosis. Understand the application of microbiology to public health and epidemiology.

Epidemiology:

SEE RESULTS TABLE

SEE HOMEWORK QUESTIONS

Exercise 9 - Ultra Violet Light: Lethal Effects

OUR GOALS:

1. To observe the effect of UV radiation on bacterial growth.

2. To learn how dimers are formed and may be repaired.

3. To observe the effect of time, covering, and plastic lid on growth.

4. To observe the resistance of endospores to UV light.

1. Ultraviolet light has germicidal effects. The most germicidal wavelength is

________________. This is useful for the sterilization of medical equipment, patient rooms and operating rooms.

2. The damage from UV light affects cells in the _______________________.

___________________________ (T=T) cause the DNA molecule to be deformed.

This prevents ___________________________ from transcribing genes to make needed proteins for cell function.

[pic]

3. Cells cope with dimer damage with the __________________________________, which produces enzymes that cut out and replace the pair.

However, if UV light exposure produces an overwhelming number of dimers, the

SOS system cannot keep up and makes errors.

4. This type of radiation causes _______________________, but also is very affective in

permanently damaging _______________________.

5.

Review Exercise 9 Ultra Violet Light: Lethal Effects

Where does UV damage occur?

How do cells cope?

+ = growth, - = no growth

|Time |S. aureus |B. subtilis |

|20sec |1. |XXXXXXXXXXXXXXX |

|40sec |2. |XXXXXXXXXXXXXXX |

|1 min |3. |10. |

|1 min, 20sec (80 sec) |4. |XXXXXXXXXXXXXXX |

|1 min, 30sec (90 sec) |5. |XXXXXXXXXXXXXXX |

|1 min, 40 sec (100sec) |6. |XXXXXXXXXXXXXXX |

|2 min |7. |11. |

|4 min |XXXXXXXXXXXXXXXX |12. |

|8 min |XXXXXXXXXXXXXXXX |13. |

|10 min |8. |14. |

|15 min |XXXXXXXXXXXXXXXX |15. |

|16 min |XXXXXXXXXXXXXXXX |16. |

|17 min |XXXXXXXXXXXXXXXX |17. |

|18 min |XXXXXXXXXXXXXXXX |18. |

|20 min |9. *Keep lid on |19. |

How much time did it take for Staph aureus cells to be killed?

How much time did it take for Bacillus subtilis cells to be killed?

What was the effect of time of UV exposure on germicidal activity?

What type of bacteria is more resistant to UV light?

Why?

Which of the bacteria that we used was more sensitive?

What happened when the plastic lid was left on?

What application would this have in a medical setting?

Exercise 10: Evaluation of Antiseptics

OUR GOALS:

1. To compare solutions for effect on bacterial growth.

2. To learn Disc Diffusion.

3. To measure the Zone of Inhibition of Growth.

1. _____________________________ are lethal to microorganisms.

2. __________________________________ kill microorganisms and are gentle enough to be applied to living tissues.

• Examples:

3. __________________________________ are applied to inanimate objects, such as walls, floors and tabletops to kill microorganisms. They damage living tissue.

• Examples _________________________________________________________

Example________________________________, used in special rooms.

4. __________________________________ reduce microbial numbers to a safe level, but do not completely eliminate all microbes.

• Example_______________________________

5. _______________________ and ________________ are important factors.

6. __________________________ and ______________ are more resistant to chemicals.

Review Exercise 10: Evaluation of Antiseptics

1.

|Solution |Where used |Examples |

| | | |

|Antiseptic | | |

| | | |

|Disinfectants | | |

| | | |

|-cidals | | |

2. Which solution was most effective? Which solution is least effective?

3. The clear area (of no bacteria growth) around the disk is called the

__________________________________________________________________.

4. We can quantitatively compare the effectiveness of the solutions by measuring the diameter of the zone of inhibition in mm.

[pic]

Exercise 5: Pure Culture Technique

OUR GOALS: 1.) To perform streak plate with the quadrant method.

2.) Learn terms: colony and CFU.

3.) Introduce selective media.

4.) Learn how to label and store plates

1. Bacteria exist in normal environments in mixed population.

2. CFU –

3. Colony -

4. Reasons to isolate bacteria:

1.

2.

3.

4.

5. TSA (tryptic soy agar) is a _____________________________________ media which

allows many different kinds of bacterial and fungal growth.

6. EMB (Eosin Methylene Blue) is _________________________ for gram negative

bacteria. That means that it does allow gram _______ to grow, but not gram _______.

EMB is also _______________________, which allows the identification of organisms by color of colony.

Review Exercise 5: Pure Culture Technique

1. What is a colony?

CFU?

2. What are the reasons for isolating colonies.

3. Results: Notable colonies:

TSA plate-

EMB plate-

Exercise 12: Protists, Fungi and Helminths

1. View slides 12-29, excluding 27 & 28.

For the test, know genus, species, common name, disease caused, structures, and microbe type: Protozoan, Fungi, Yeast, Round worm, Flat worm.

2. General taxonomy

|Domain: Bacteria |Domain: Archaebacteria |Domain: Eukaryotes |

| | | |

|Kingdom: Bacteria |Extremophile bacteria |Kingdom: Formerly Protista |

| | |Entamoeba histolytica |

|Phylum: Bacteria | |Trypanosoma gambiense |

|(All bacteria we have studied in slides | |Trichomonas vaginalis |

|3-11, but not included in Test 2 material.)| |Plasmodium sp. |

| | | |

| | |Kingdom: Animalia |

| | |Phylum: Nematoda |

| | |Ascaris lumbricoides |

| | |Ancylostoma caninum |

| | |Trichinella spiralis |

| | |Enterobius vermicularis |

| | | |

| | |Phylum: Platyhelminthes |

| | |Clonorchis sinensis |

| | |Fasciola hepatica |

| | |Taenia pisiformis |

| | | |

| | | |

| | |Kingdom: Fungi |

| | |Phylum: Ascomycota |

| | |Saccharomyces sp. |

| | |Aspergillus sp. |

| | |Penicillium sp. |

| | |Peziza sp. |

| | | |

| | |Phylum Zygomycota |

| | |Rhizopus stolonifera |

| | | |

| | |Phylum Basidiomycota |

| | |Coprinus sp. |

3. PROTOZOA

1. Life stages _____________________ and ________________________.

(dormant) (growing & reproducing)

2. All reproduce ______________________________. (bianary or multiple fission)

Some reproduce ____________________________. (hermarphroditic)

Some reproduce both ways.

TERMS & STRUCTURES: Nucleus, flagella, red blood cells, cyst, trophozoite

4. WORMS

All worms are in the Kingdom __________________________________.

Worms are either _____________________ or _____________________.

Clinically, worms are identified by their __________________________.

TERMS & STRUCTURES: scolex, hooks, suckers, gravid, proglottid, helminth, ovaries, testes

5. FUNGI

Distinguishing characteristics of fungi are:

1. ________________________________________________________

2. ________________________________________________________

• Hyphae may be ________________, which have cross walls between nuclei in

hyphae, or be _____________________.

• Fungi are identified by _________________________ in their reproduction cycles.

• Fungi can reproduce _______________________ or ______________________.

TERMS & STRUCTURES: Sexual spores: zygospore, ascospores, basidiospores. Asexual spores: sporangiospores, conidia, yeast. Other Structures: gills, stalk, hyphae, septate, nonseptate.

6. YEASTS

Yeasts are FUNGI that lack ____________________.

Yeasts are __________________________, while molds are multicellular.

We will see yeast reproducing by _______________________________.

Yeasts may have _________________________, which are elongated cells.

TERMS AND STRUCTURES: budding, pseudohyphae.

Exercise 13: Effect of Temperature on Growth

OUR GOALS:

1. To learn how temperature affects bacteria metabolism.

2. To categorize bacteria based on temperature range of growth.

1. Bacterial growth is sensitive to environmental factors, such as: _______,

__________, ___________________, and _______________________.

2.

__________ = refrigerator temperature

________________ = room temperature

__________ = body temperature

__________ = above body temperature

3. Temperature affects bacterial metabolism because of enzyme _______________ and

_____________________.

[pic]

Temperature increase = collision increase (substrate and enzyme)

4. Processes that enzymes function in (some examples): movement, reproduction, recombination, digesting food, producing ATP, repairing cellular damage, producing structures such as capsules or endospores.

Exercise 14: Transformation in Bacteria

OUR GOALS:

1. To see how microbiology is used in biotechnology.

2. To observe bacterial transformation by uptake of DNA from surroundings.

3. Relate this process to antibiotic resistance.

1. In BIOTECHNOLOGY,

Genes isolated ( function determined ( copied ( vector ( expression ( collected

2. Current use includes:

Human insulin DNA expressed in bacteria

Genetically modified organisms: foods

Vaccines

[pic]

xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx

_________________ = Protein that glows green

_________________ = ampicillin antibiotic resistance

_________________ = arabinose promoter to break down sugar for bacteria to grow

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