LABORATORY

MICP

Bachelor of Science in Nursing: MICP LAB

LAB MODULE LAB UNIT WEEK

4 1 4

I. Physical & Chemical Sterilization

II. Immunity, Hypersensitivity and Vaccines

 Implement a study habit to read and comprehend the following prior to online class proper:
unit objectives; lab module and lab learning videos.
 Actively search for unfamiliar medical terminologies and relate to discussions.
 Establish effective teacher- student interactions through participation in the synchronous
online class discussion. through LMS discussion board or through Online Lab Class chat box. Ask
relevant questions.
 Answer and submit lab unit tasks online if there is any.
 For additional direction read study guide prior to class proper

At the end of this unit, the students are expected to:

1. Identify the different physical and chemical methods of sterilization.

2. Differentiate innate from adaptive immunity, antigen from immunogen
3. Apply the concept of immunogenicity properties
4. Discuss disorder that may arise because of immunity
5. Elaborate the production of vaccines

LAB GUIDE

This Module was based on Microbiology & Parasitilogy Textbook & Lab
Manual by Drs. Bartolome and Quiles. Chapter 9. After that, on pages
131-132, answer the Self-Assessment Questions.

1
 I. PHYSICAL AND CHEMICAL METHODS OF STERILIZATION

 Sterilization is a technique that uses physical or chemical methods to eradicate or eliminate all
kinds of microbial life in health-care facilities.
 Disinfection is the process of destroying most microbiological forms in a nonliving material, except
saprophytes and bacterial endospores.
 Antisepsis is the application of chemical agents to living tissues (skin) to prevent the spread of
microbes by inhibiting or killing their growth.
Bactericidal or germicidal an agent that kills the bacteria.
 Bacteriostatic agents are substances that can impede bacteria's proliferation without necessarily
killing them.
 Agents that kill spores, fungi, and viruses are known as sporicidal, fungicidal, and viricidal.

PHYSICAL METHODS OF STERILIZATION

HEAT - universally applicable method of sterilization. Rate of killing is expressed in thermal death time

1.MOIST HEAT - preferred over dry heat because of its more rapid killing action.
> Steam under pressure (autoclaving)- Most efficient method of sterilization, it can destroy all
microbial forms
> Boiling- inv. utilizing temp. of 80 -100 0 C ,Destroys vegetative forms of bacteria but not endospores
> Fractional sterilization (Tyndallization) - the material is exposed to live steam for 3o mins for 3
consecutive days .
Vegetative forms are destroy not endospores
> Pasteurization - destroying disease-producing organisms in milk and milk products and other
beverages

2.DRY HEAT - – used to sterilize materials in enclosed tubes, oils, jellies, powders & glassware( test tubes
and petri dish)
> Hot air Oven - used to sterilization of powders, ointment, other glassware that cannot be sterilized
using autoclave.
> Incineration - aimed at burning the organisms into ashes (garbage disposal)
> Open Flame - burning the organism into ashes Ex. Bunsen burner and alcohol lamps

DESSICATION - - deprive the microorganisms of moisture .Used in food preservation (dried fish and fruits)
FREEZING - Lyophillization (freeze-drying) organism is rapidly frozen, then dehydrated in high vacuum and
stored in a vacuum sealed container ….. Ex. Large laboratories for preserving microorganisms
FILTRATION – form of mechanical sieving .Microorganisms are not killed, but merely separated from the
fluid

RADIATION –
1.Ultraviolet light (UVL) –most effective wavelength of UVL is 240-280 nm .Its application is limited only to
sterilization of small enclosed areas like operating rm & hospital wards bec/ of its poor penetrating
ability

2
 2.Ionizing radiation – formation of free radicals that will chemically interact w/ proteins & nucleic acids
resulting in cell death. Potentially hazardous to human cells.
OSMOTIC PRESSURE – based on principle of osmosis .For preservation of foods in syrup & meats in brine.

CHEMICAL METHOD OF STERILIZATION

Factors that can affect efficacy of chemical agent:

Concentration of chemical agent - -higher concentration is bactericidal -lower concentration is bacteriostatic
Time - – the longer time of exposure the better the killing action
Temperature - – higher temperature speeds up the rate of chemical reaction
Nature of surrounding medium- the pH of the medium & the presence of pus or blood decreases efficiency of
chemical agent.
Nature of organism- refers to ability of bacteria to produce endospores w/c are resistant to chemical agents

MECHANISMS OF ACTION

Damage to the cell membrane - can cause smaller molecules to leak out the bacterial cell and interfere the active
transport and energy metabolism within the cell .

1.Surface active agents - decrease surface tension of cell membrane lipids

> Cationic agents - quaternary ammonim compound (zephiran) that are effective at alkaline Ph
> Anionic agents- - like soaps & detergent remove dirt through EMULSIFICATION most effective at acidic pH

2. Phenolic compounds-
> Phenol - no longer used as disinfectant, its toxic to human cells
> Cresol - more safer than phenol (Lysol)

3. Alcohol - disorganize the lipid structure and cause denaturation of cellular proteins
> ethyl alcohol - bactericidal and remove lipid from the skin surface
> isopropyl alcohol – has greater bactericidal activity and less volatile but exerts greater toxic effects (narcosis
due to inhalation of fumes)
> benzyl alcohol- used as a preservative

MODIFICATION OF FUNCTIONAL GROUPS OF PROTEINS AND NUCLEIC ACIDS

1. HEAVY METALS - – damage the enzyme activity of bacteria
a. Mercurials ( merthiolate or mercurochrome) –ARE BIOCIDAL & USED AS ANTISEPTICS
b. Silver compounds are bactericidals - (1% silver nitrate –prophylaxis for ophthalmia neonatrum)
(silver sulfadiazine – tx for burn wounds)

2. HALOGENS - – bactericidal oxidizing agents that cause the inactivation of enzymes

a. Iodine – CONSIDERED AS BEST ANTISEPTIC, bec. of its sporicidal bactericidal, fungicidal, amoebicidal,
viricidal.
b. Chlorine – used in the treatment of water.Also a common household disinfectant
c. Hydrogen peroxide - ->a weak antiseptic and used for cleaning wounds

3. ALKYLATING AGENTS
a. Formaldehyde - – sporicidal and commercially available as FORMALIN
b. Glutaraldehyde - – sporicidal and used as cold sterilant in sterilizing medical equipment.
c. Ethylene oxide - sporicidal and used in gaseous sterilization of heat-sensitive materials equipment

3
 II. IMMUNITY

Image:technologynetwoks

 The immune system is a complicated network of cells and proteins that fights infection in the
body.
 The immune system keeps track of every infection (microbe) it has ever eliminated so it can
rapidly recognize and eliminate the microbe if it re-enters the body.

LAB TASKS

Application: Innate Immunity and Adaptive Immunity. Part I

Instruction: Choose 1 from the following options: COVID-19, Hepatitis B virus, HPV, Measles virus,
Cholera,Tetanus Clostridium tetani or from other vaccine-preventable diseases.

What natural defenses do you have against this infection? What role does your adaptive immunity play
in helping and protecting you against it. Give a brief group report.
__________________________________________________________________________________

4
  Immunity is a complex biological system with the ability to recognize and accept what belongs
to the self, as well as to recognize and reject what does not belong to the self (non-self).
 Immune response means antigenic challenge leads to activation of either B-lymphocytes or T-
lymphocytes.These foreign substances may be: Immunogenic, Antigenic. Properties Of Antigen
(Foreign Substances) To Be Immunogenic are:
 A. Foreignness. Identifies the substances as being foreign (non-self)
 B. Chemical Composition. Its either proteins or large polysaccharides molecules. rarely
lipids.
 C. Molecular size (molecular weight). The larger the molecule's molecular weight, the
better it will work as an antigen.
 D. Chemical Complexity. The more complex chemical structure is more potent
immunogen
 E. Dose of Immunogen. If tiny in amount, very poor immune responses could be induce
 F. Genetic Constitution of the Host
 G. Route of Entry of Immunogen
 You may use this link as guide:
https://immunologyinfo.weebly.com/antigen.html

LAB TASKS

Search and Analyze (Part II)

Our immune system is successfully elicited by the Corona virus.

Instruction: Differentiate the two: Antigen vs Immunogen. Give a brief group report about the
properties of Corona virus as an immunogen.

____________________________________________________________________________________

 Abnormalities of the immune system can lead to allergic diseases, immunodeficiencies and
autoimmune disorders.
 For details, you may use this suggested links:
https://www.hopkinsmedicine.org/health/conditions-and-diseases/disorders-of-the-immune-
system

5
 https://knowablemagazine.org/article/health-disease/2020/what-cytokine-storm

https://www.health.harvard.edu/diseases-and-conditions/covid-19-basics

LAB TASKS

Search and Analyze (Part III)

When we get sick we trust our immune system to protect us, but what happens when that system
goes awry and ends up killing patients?

Instruction: What is Cytokine Storm, and how does it work? What happens during the immune
response to the Corona virus, which results in a high number of deaths?

Analyze a give a brief report. Use ppt.

________________________________________________________________________________

 Our immune systems are built to recall things. We are usually protected against a disease for years,
decades, or even a lifetime after receiving one or more doses of a vaccination. Rather than treating
a disease after it has developed, vaccines prevent us from being sick in the first place.
 To show you in video format, use these suggested links:

https://www.youtube.com/watch?v=-92HQA0GcI8

https://www.youtube.com/watch?v=J0SvutsCHrU

https://www.youtube.com/watch?v=W-Zi5TitJzk

LAB TASKS

Search and Understand (Part IV)

How are vaccines made? Understand the shared videos and make a brief group report.

____________________________________________________________________

You are doing great!

6
Now let us answer your assignment:The Self-Assessment Module in pages 131 to 132.

Now, Let’s answer the Self-Assessment pages 131

-132 of the textbook Microbiology and Parasitology
A Textbook and Laboratory Manual for the Health
Science, 2nd Edition by Dr. Bartolome, Dr. Quiles

Here is a list of external links used for this module. Copy & paste the following link into your browser to gain access:

Video
Part 2 Immunogen and Antigen https://labpedia.net/elementary-immunology/chapter-3-
immunogen-and-antigen/

Part 3 What is a cytokine storm? https://www.hopkinsmedicine.org/health/conditions-

and-diseases/disorders-of-the-immune-system

https://knowablemagazine.org/article/health-
Covid - 19 Basics disease/2020/what-cytokine-storm

https://www.health.harvard.edu/diseases-and-
conditions/covid-19-basics
Part 4 Covid- 19 vaccine https://www.youtube.com/watch?v=-92HQA0GcI8

Vaccine manufacture: it’s complicated https://www.youtube.com/watch?v=J0SvutsCHrU

Sanofi – Making vaccines https://www.youtube.com/watch?v=W-Zi5TitJzk

Congratulations! You've come this far, so keep going for your goals.

This week's most important terminologies are given below. Remember to understand,
learn, and stay safe while preparing for your future as RNs.

TERMINOLOGIES

7
 Acellular vaccine: A vaccine made up of partial cellular material rather than whole cells.
 Adjuvant : A vaccine component that boosts the immune response.
 Anaphylaxis: An allergic reaction that occurs quickly and severely. (e.g. food or drugs). Symptoms
of anaphylaxis include breathing difficulties, loss of consciousness and a drop in blood pressure.
This condition can be fatal and requires immediate medical attention.
 Antitoxin: solution of antibodies against a toxin. Antitoxin can be derived from either human (e.g.,
tetanus immune globulin) or animal (usually equine) sources (e.g., diphtheria and botulism
antitoxin). Antitoxins are used to confer passive immunity and for treatment.
 Booster shots:Additional vaccination doses are required on a regular basis to "boost" the immune
system. For example, the tetanus and diphtheria (Td) vaccine which is recommended for adults
every ten years.
 Breakthrough infection: Development of a disease despite a person’s having responded to a
vaccine.
 Combination vaccine: A product containing components that can be divided equally into
independently available routine vaccines.
 Community immunity: When a large enough percentage of a population is immune to an
infectious disease. (through vaccination and/or prior illness) to make its spread from person to
person unlikely. Even individuals not vaccinated (such as newborns and those with chronic
illnesses) are offered some protection because the disease has little opportunity to spread within
the community. Also known as herd immunity.
 Conjugate vaccine: To boost the efficiency of a vaccination, two compounds (typically a protein
and a polysaccharide) are joined together.
 Efficacy rate: A measure used to describe how good a vaccine is at preventing disease.
 Investigational vaccine: A vaccine that has been approved by the Food and Drug Administration
(FDA) for use in clinical trials on humans. However, investigational vaccines are still in the testing
and evaluation phase and are not licensed for use in the general public.
 Potency: A measure of strength.
 Quarantine: The isolation of a person or animal who is suspected of having a disease in order to
prevent further spread of the disease.
 Seroconversion: Development of antibodies in the blood of an individual who previously did not
have detectable antibodies.
 Serology: Measurement of antibodies, and other immunological properties, in the blood serum.
 Serosurvey: Study measuring a person’s risk of developing a particular disease.
 Strain: A specific version of an organism. Many diseases, including HIV/AIDS and hepatitis, have
multiple strains
 Teratogenic: Of, relating to, or causing developmental malformations.
 Thimerosal: Thimerosal is a mercury-containing preservative used in some vaccines and other
products since the 1930’s. There is no convincing evidence of harm caused by the low
concentrations of thimerosal in vaccines, except for minor reactions like redness and swelling at
the injection site.
 Waning Immunity: The loss of protective antibodies over time.

8
 Bartolome and Quiles. (2020) Microbiology and Parasitology: A Textbook and Laboratory
Manual for the Health Sciences. 2nd Edition. C&E Publishing House. Quezon City. QR65. B37 2020
Tortora, G.J. (2014) Microbiology. Pearson: Singapore. 616.9041 T63 2014
Engelkirk, Paul G. (2019). Burton's microbiology for the health sciences, 8th ed. . Philadelphia :
LWW.616.01 E3 2007
www.cdc.com
www.googlesearch
Conditions and Disease of Immune System https://www.hopkinsmedicine.org Accessed January 28,
2022
Covid - 19 Basics. https://www.health.harvard.edu/ Accessed January 28, 2022
Cytokine Storm https://knowablemagazine.org Accessed January 28, 2022
Covid- 19 vaccine https://www.youtube.com/watch?v=-92HQA0GcI8 Accessed January 28, 2022
Immunogen and Antigen www.Labpedia.net.com Accessed January 28, 2022
Sanofi – Making vaccines https://www.youtube.com/watch?v=W-Zi5TitJzk Accessed January 28, 2022
Vaccine & Immunizations Glossary https://www.cdc.gov/vaccines/terms/glossary.html January 28, 2022
Vaccine manufacture: it’s complicated https://www.youtube.com/watch?v=J0SvutsCHrU Accessed
January 28, 2022
/AVG