Name: Jaiy Hingco Date Performed: 9/2/19

Section: 2 Date Submitted: 9/8/19

Group No. 1
Laboratory Exercise No. 6
Disinfectants
Results

Table 1. Effect of Various Disinfectants in bacterial cultures of Escherichia coli

Disinfectant Concentration Growth after time of exposure (minutes)

Control
5 10 15 C

Sodium hypoclorite 5% 3 2 1 4
0.05% 3 2 1 4
Alcohol Absolute 1 3 2 4
Hydrogen peroxide 3% 3 2 1 4
Lysol 1% 1 3 2 4
Mouthwash NaF 0.05% 2 3 1 4

Table 2. Effect of Various Disinfectants in bacterial cultures of Staphylococcus aureus

Disinfectant Concentration Growth after time of exposure (minutes)

Control
5 10 15 C

Sodium hypoclorite 5% 3 2 1 4
Alcohol Absolute 1 3 2 4
Hydrogen peroxide 3% 2 1 3 4
Lysol 1% 2 1 3 4
Mouthwash NaF 0.05% 2 3 1 4

Growth Rankings Growth Description

1 Least growth
2 Less growth
3 More growth
4 Most growth
 Name: Jaiy Hingco Date Performed: 9/2/19
Section: 2 Date Submitted: 9/8/19
Group No. 1
Laboratory Exercise No. 6
Disinfectants
Study Questions

1. Define disinfection. Compare/contrast it with sterilization, antisepsis and bacteriostasis.

Disinfection is a process of killing, inhibition, removal or many or all pathogenic microorganisms, except bacterial
spores, on inanimate objects, while sterilization is the killing of all forms of microbial life in an object or habitat.
Unlike disinfection, antisepsis is the practice of using antiseptics to eliminate microbes that cause disease, prevent
infection or sepsis in living tissues, while bacteriostasis is the inhibition of the growth and reproduction of bacteria,
especially by the action of a chemical agent (Rutala, et al.,2016).

2. Give the modes of action of the different antiseptics/disinfectants used in the activity.

The mechanism of action of mouthwash involves bacterial cell wall destruction, bacterial enzymatic
inhibition, and extraction of bacterial lipopolysaccharides; Sodium hypochlorite involve biosynthetic
alterations in cellular metabolism and phospholipid destruction, best used in water sanitation;
Hydrogen peroxide oxidizes the microbial cell wall to attacks essential cell components like the
DNA. Alcohol and Lysol destroys the cell membrane and denatures cell wall proteins, allowing it to
be effective in dissolving outer membrane of (G -) bacteria (Choudhary, 2015).

3. Which microbial forms are most resistant to disinfectants?

Endospores are definitiely the most resistant to disinfectants as it can survive without nutrients and is
even resistant to ultraviolet radiation, desiccation, high temperature, extreme freezing and chemical disinfectants,
due to its DPA (dipocolinic acid) and spore coat. This is followed by mycobacteria, then Gram negative bacteria
(Antimicrobial Disinfectants, 1994).

4. What is an iodophor? What is its value?

Iodophors are intermediate-level disinfectants containing iodine combined with surfactant, effective against
vegetative bacteria, enveloped viruses, fungi, and some mycobacteria. It is used as an antiseptic for the prevention
and treatment of wounds. It is used in brewing and dairy industries as sanitizer as iodine oxidizes the hydrogen-
sulfur bonds in the amino acids, preventing microorganisms to synthesis peptide and leading to its death (Larson
and Morton, 1991).

References:

Antimicrobial Disinfectants. (1994). Retrieved from https://www.ncbi.nlm.nih.gov › pubmed

Choudhary, K. (2015). Mode of Action of Disinfectants. Retrieved from https://www.pharmaguideline.com/2015

Larson, E. L. & Morton, H. E. (1991). Disinfection, Sterilization and Preservation. Alcohols, 4th Edition

Rutala, W. et al. (2016). Guideline for Disinfection and Sterilization in Healthcare Facilities. Retrieved from
https://www.cdc.gov/
Name: Jaiy Hingco Date Performed: 9/2/19
Section: 2 Date Submitted: 9/8/19
Group No. 1

1. Based on your results, which chemical is most effective against E. coli? Which is least effective against E. coli?

2. Which is most effective against S. aureus? Which is least effective against S. aureus?

3. Why do microorganisms differ in their response to disinfectants?

4. Which chemical would you choose if you wanted maximum effectiveness against both E. coli and S. aureus?

5. What do you think are the factors influenced the activity of a disinfectant? How?

6. Did you find the mouthwash you tested to be as effective as the other disinfectants included in this exercise? Explain any
difference you observed.
 ODEZZA ANSWERS DON’T PRINT
Laboratory Exercise No. 6
DISINFECTANTS

Table 1. Effect of disinfectants in bacterial cultures of Escherichia coli

Disinfectant Concentration Growth after time of exposure (minutes)

Control
5 10 15

Sodium hypoclorite 5% moderate (1) moderate (3) moderate (2) extensive (4)
Alcohol 70% moderate (3) moderate (2) minimal (1) extensive (4)
Hydrogen peroxide 5% moderate (3) moderate (3) moderate (3) extensive (4)
Lysol 5% moderate (3) moderate (2) moderate (1) extensive(4)
Mouthwash NaF- 0.02%
(Listerine Healthy White) OH- 8% moderate (2) extensive (3) extensive (4) moderate (2)

Table 2. Effect of disinfectants in bacterial cultures of Staphylococcus aureus

Disinfectant Concentration Growth after time of exposure (minutes)

Control
5 10 15

Sodium hypoclorite 5% moderate (2) moderate (2) moderate (3) extensive (4)
Alcohol 70% extensive (3) moderate (2) extensive (4) moderate (1)
Hydrogen peroxide 5% minimal (1) minimal (2) moderate (3) extensive (4)
Lysol 5% moderate (3) moderate (2) minimal (1) extensive (4)
Mouthwash NaF- 0.02%
(Listerine Healthy White) OH- 8% moderate (2) extensive (3) extensive (4) moderate (1)
Legend:
Growth Rankings Growth Description
1- Least growth No growth
2- Less growth Minimal
3- More growth Moderate
4- Most growth Extensive

Answer the following questions using the results obtained from the exercise

1. Based on your results, which chemical is most effective against E. coli? Which is least effective against E.
coli? Alcohol is the most effective disinfectant against E. coli while mouthwash, particularly Listerine
Healthy White is the least effective.

2. Which is most effective against S. aureus? Which is least effective against S. aureus? Lysol is the most
effective disinfectant against S. aureus while Listerine Healthy White is the least effective.

3. Why do microorganisms differ in their response to disinfectants?

Microorganisms have unique characteristics that differ from one genus to another. Disinfectants attack
 specific target in the cell which not all microorganisms may possess. Moreover, some of them have
enzymes that are capable of removing toxins inside their membrane making them resistant to disinfectants.

4. Which chemical would you choose if you wanted maximum effectiveness against both E. coli and S.
aureus? Lysol is an effective disinfectant for both gram positive and gram negative bacteria based on the
result of the experiment. Aside from the disinfectants used in the experiment, researches over the internet
shows that chlorhexidine, a positively-charged molecule that binds to negatively-charged cell wall and
disrupts the cell membrane, is 100% effective against both type of bacteria.

5. What do you think are the factors that influenced the activity of a disinfectant? How?
The number of microorganisms, the time of exposure, temperature and pH are the factors that influence the
activity of disinfectant towards microorganisms. The larger the number of microbes, the more time a
disinfectant need to destroy all of them which is more effective in a favorable condition like high
temperature and pH not suitable for that particular microorganisms.

6. Did you find the mouthwash you tested to be as effective as the other disinfectants included in this
exercise? Explain any difference you observed.
Listerine Healthy White is not as effective as the other disinfectants in eradicating both gram negative and
gram positive bacteria. It shows inconsistency, at the first 5 minutes it is able to lessen bacterial growth but
as time goes by there is a drastic increase in the bacterial growth which is noticeably higher compared to
the control group.

Study Questions

1. Define disinfection. Compare/contrast it with sterilization, antisepsis and bacteriostasis.

Disinfection means destroying harmful microorganisms in contact with this chemical. This is
closely related but not synonymous to antisepsis which involves not only the distruction of
pathogenic microorganisms but also inhibition of its growth and multiplication; to sterilization
which is define as the removal of microorganisms regardless if their pathogenic or not; and to
bacteriostasis which is an inhibition of the growth of bacteria without destruction.

2. Give the modes of action of the different antiseptics/disinfectants used in the activity.

Sodium hypochlorite and hydrogen peroxide both oxidizes the cell wall of microorganisms and
attacks essential cell components including the DNA. Alcohol and Lysol destroys the cell
membrane and denatures the protein of the cell, making it more effective in dissolving the outer
membrane of gram negative bacteria. Listerine mouthwash contains active ingredients including
hyroden peroxide, sodium fluoride and alcohol which acts the same way but not as effective as the
chemicals mention above.

3. Which microbial forms are most resistant to disinfectants?

Bacterial endospore which can survive for thousands of years dormant are the most resistant due to
its protective covering of spore coat. It is followed by mycobacterium that contains mycolic acid
layer not soluble to most chemicals.

4. What is an iodophor? What is its value?

Iodophor is a group of disinfectants containing iodine in combination with a surfactant. It is used in

brewing and dairy industries as sanitizer since iodine oxidizes the hydrogen-sulfur bonds in the
amino acids, preventing microorganisms to synthesis peptide and leading to its death.
References:

 Larson, E. L. & Morton, H. E. (1991). Disinfection, Sterilization and Preservation. Alcohols, 4th Edition
 UC Davis Viticulture and Enology (April 3, 2018). The Regents of the University of California, Davis
campus. Retrieved from: https://wineserver.ucdavis.edu/