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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
Chapter 1
INTRODUCTION
the clinical specimen. Its purpose is to see if the microbe can ferment the
Phenol Red, as the indicator for organisms that can ferment Mannitol
which is the major and differential ingredients of MSA and lastly the
organism that cannot tolerate the high saline levels (Acharya, 2013).
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
or absence of oxygen. The bacteria tend to infect the skin often causing
abscesses. However, the bacteria can travel through the bloodstream and
infect almost any site in the body, particularly heart valves and bones. As
that is also a good source of protein and vitamins and mineral to humans.
Since Brassica oleracea var. botrytis is generally accepted that its extracts
the Brassica oleracea var. botrytis for the growth of Staphylococcus aureus.
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
STATEMENT OF HYPHOTHESES
Positive bacteria.
differentiate microorganisms.
responsible for the bacterial growth were also not included in this study.
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
Cauliflower
Mannitol
for example, contains 3 grams of Mannitol per every 100 grams of weight.
butternut squash and sweet potato (Muir et al., 2009). According to Smith,
folic acid supplements, vitamins, candy and baked goods. Mannitol also
higher. For example: cauliflower has about 2.6 grams of mannitol per 100
grams; mushrooms have about 2.6 grams of mannitol per 100 grams;
snow peas have about 1.2 grams of mannitol per 100 grams; peaches have
IBS and avoiding foods containing mannitol can help alleviate the
harsh, salty environment. So Staph grow well in this media. This growing
the pH indicator in MSA from red to bright yellow. Pathogenic Staph, such
Mannitol Salt Agar, their wastes turn the MSA a bright yellow color. By
epidermidis (a.k.a. Staph epi), the normal flora that grows on human skin,
does not ferment mannitol. When Staph epi grows on Mannitol Salt, the
of 32°C.
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
red.
Staphylococcus aureus
exponential manner: with each division cycle (generation), one cell gives
rise to 2 cells, 4 cells then 8 cells, then 16 and 32 and so forth. There is a
or in the nose. It can cause a range of mild to severe infections and may
However if it enters the body through a cut in skin, it may cause death
bacteria commonly found on the skin and hair as well as in the noses and
percent of healthy people and are even more common among those with
poisoning when a food handler contaminates food and then the food is not
ranging from a mild skin infection to blood stream infections and deep
metastatic foci increases the risk of mortality and raises the stakes for
2009).
Staphylococcus epidermidis
heart valves, artificial lenses and orthopedic implants. The infection starts
ability to form biofilms. Cells in this mode of growth are inherently more
morbidity and health care costs, many efforts are being made to develop
of the microbiota of human and animal skin and mucosa. Over a period of
culture media, and its potential and culturing fungi was evaluated for cost
gelling ability. The cassava starch alone cannot be use for culture media
legume seeds that have found to have a good protein source for nutritional
purposes. Different protein source is also use in the study for growing a
bacteria such as E.Coli, Bacillus sp., Klebsiella sp., Staphylococcus sp., and
Pseudomonas sp.
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
Control
Positive:
Mannitol Salt Agar
Staphylococcus aureus
Negative:
Staphylococcus Growth of bacteria
epidermidis
Experimental group
Cauliflower Agar
Effectiveness of
Cauliflower as substitute
to Mannitol MSA
Chapter 2
applied in this study. The manner how the study was designed and
RESEARCH DESIGN
plate with the cauliflower powder substitute for mannitol where the
observed. The Mannitol Salt Agar plate serve as the control group assessed
RESEARCH SETTING:
Cabanatuan City Public Market. It was bought only in one stall at the same
Santo Tomas, España, Manila and also the pulverization process was held
City Public Market and was cut into small pieces before it was used in the
experiment.
5)
Culture media
7.5% NaCl, 1% Mannitol 1.5% Agar, and Phenol Red, the pH is adjusted
Sampling Procedure:
Cabanatuan City Public Market within one day (morning). The samples
were cut into very thin slices, air dry and put in the blender until finely
powdered.
General Procedure:
weighed using a triple beam balance. It was cut into a smaller pieces and
air dried. A grinder to turn it into a powdered form and was kept in an
Phenol Red 75 grams of NaCl and 1000ml distilled water. The prepared
agar mix was gently heated until it dissolved and then autoclaved. (Figure
9)
dish and allowed to stand for a minute, then it was refrigerated at 4°C.
Researchers did a triplicate sample of each Agar Plate. (Figure 10, Figure
11)
epidermidis were inoculated from the Nutrient Agar. Using the overlapping
streak plate method, the bacteria from the pure culture was streaked to
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
the prepared Cauliflower agar and Mannitol Salt Agar using a disposable
INCUBATION
After streaking of the bacteria into the prepared agar plates, it was
observation and monitoring of the agar plates were done in 12th and
(Figure 13)
BIOCHEMICAL TESTING
the species that grow on the agar plates. Researchers first describe the
morphology on the agar plate then perform the Gram Staining to identify
container for the agar while heating. Biosafety Cabinet was used as the
suitable for heating liquids. It was also used for the preparation of
of distilled water, Hot plate was used to heat and dissolve the agar,
Incubator was the device used to grow and maintain the bacterial culture
for the bacteria to grow, Inoculating loop was used to transfer and streak
an inoculum from the cultured bacteria to another agar plate, Petri dish A
flat dish with a lid which held the prepared solid agar, It was used for
aureus This bacteria was used to grow in Cauliflower Agar for Positive
contained the powdered cauliflower, Stirring rod was used to stir the agar
while being heated. BBL™ CRYSTAL™ ID KIT was used to determine if the
pretty much as they sound — they describe situations. There are three
ETHICAL CONSIDERATIONS
laboratory tools and equipment were also done. Data and document report
were all in accordance to what have been observed during the period. There
harmed during the conduct of this study. The internal validity is also
CHAPTER 3
This chapter presents and discusses the data collected based from
I. CONTROL GROUPS
Agar 2 3 A
1- Without Growth
2- Few Growth
3- Moderate Growth A- Yes
4- Heavy Growth B- No
Table 1.1 shows the result of trial under the controlled conditions
growth through time in the Mannitol Salt Agar and a change in color from
red to yellow.
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
Agar 2 3 B
Table 1.2 shows the result of the trial under the controlled conditions
Agar 2 2 B
1- Without Growth
2- Few Growth
A- Yes
3- Moderate Growth
B- No
4- Heavy Growth
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
Table 2.1 shows the result of the trial under the controlled
aureus onto the Cauliflower Agar It shows that Staphylococcus aureus has
the same growth through time and it does not show any color changes at
Agar 2 2 B
1- Without Growth
2- Few Growth A- Yes
3- Moderate Growth B- No
4- Heavy Growth
This Table 2.2 shows the result of the trial under the controlled
does not show any changes in its color at 12th and 24th hours of
observation.
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
grow a little slower and 48 hours might be the appropriate time to check
for growth.
I. Morphology
R1 1.0-1.5 mm 0.8-1.2 mm
diameter, diameter
R2 1.0-1.5 mm 0.8-1.2 mm
Colonies
R3 1.0-1.5 mm 0.8-1.2 mm
Colonies
Cauliflower Agar
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
Cauliflower agar for the identification of the colonies. As the table shows,
the colonies that grow on the agar are Staphylococcus aureus and
Cauliflower Agar Plate. This shows that the organism on the plate are a
Gram Positive bacteria have a very thick cell wall made of protein
called peptidoglycan. These bacteria retain the crystal violet dye (chemical
AGAR
Table 3.3 shows the species of bacteria from the Cauliflower agar.
Staphylococcus epidermidis.
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College of Nursing and Allied Medical Science
FIGURE 16. 12th hours A) (Left) Mannitol Salt Agar streak with
Staphylococcus aureus
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
FIGURE 17. 12th hours C) (Left) Mannitol Salt Agar streak with
Staphylococcus epidermidis.
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
FIGURE 18. 24th hours A) (Left) Mannitol Salt Agar streak with
Staphylococcus aureus.
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
FIGURE 19. 24th hours C) (Left) Mannitol Salt Agar streak with
Staphylococcus epidermidis.
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
(Right)
Chapter 4
presents the key points of this study. The conclusion answers the
SUMMARY
CONCLUSION
grow.
RECOMMENDATIONS
their media.
4. Finding out the components that could promote a growth and ability
agar.
APPENDICES
Greetings!
We are students from Wesleyan University-Philippines, and currently conducting our
undergraduate research study entitled “Cauliflower as a substitute for Mannitol Salt
Agar, a differential media for isolation of Staphylococcus aureus”. As part of our
research, we need a bacterial strain of Staphylococcus aureus and Staphylococcus
epidermidis that serves as ourr positive and negative control.
We are hoping for your response regarding this matter. Thank you and God bless!
Respectfully yours,
Sir:
Greetings!
We are students from Wesleyan University-Philippines, and we are currently conducting our
undergraduate research study entitled “Cauliflower as a substitute for Mannitol Salt Agar, a
differential media for isolation of Staphylococcus aureus”. In the line with this, we are tasked to
certify our plant samples under the National Museum Botany Department. This is to guarantee the
authenticity of the sample we are about to use in our experimentation.
An attachment to this letter includes images of our sample and the sample to be submitted to the
National Museum Botany Department.
We are hoping for your response regarding this matter. Thank you and God bless!
Respectfully yours,
Andres, Kristel Joy
Dela Paz, Crisha Darryale
Dela Paz, Cristian Deo
Miralles, Marjorie Noted by:
Sigue, Rachelle Sharey R. Capati, RMT
Teano, Cristiana (Research Adviser)
(Researchers)
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WESLEYAN UNIVERSITY-PHILIPPINES
College of Nursing and Allied Medical Science
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