[WESLEYAN UNIVERSITY – PHILIPPINES

COLLEGE OF NURSING AND ALLIED MEDICAL SCIENCES

DEPARTMENT OF NURSING] May 23, 2018

CAULIFLOWER AS A SUBSTITUTE
INGREDIENT FOR MANNITOL IN MANNITOL
SALT AGAR
Andres, Kristel Joy; Dela Paz Crisha Darryale; Dela Paz Cristian Deo; Miralles, Marjorie; Sigue, Rachelle; Teaño, Cristianaa, Sharey Capati, RMTb
a
Bachelor of Science in Medical Technology, College of Nursing and Allied Medical Sciences, Wesleyan University -
Philippines, Cabanatuan City, Nueva Ecija

Andres.kristeljoy26@gmail.com, Delapaz.crishadarryale13@gmail.com, delapazcristiandeo@yahoo.com,

Miralles.marjorie13@gmail.com, sigue.rachelle11@gmail.com, Cristianateano@yahoo.com

ABSTRACT: This study generally aimed to test or determine the effectiveness of Cauliflower as a substitute mannitol
ingredient for Mannitol salt agar to differentiate Staphylococcus aureus to other Gram positive bacteria. Quasi-
experimental research design was the methodology utilized in the study. The Descriptive analysis revealed that the use of
a pulverized cauliflower allowed the growth of Staphylococcus aureus and Staphylococcus epidermidis, but as a substitute
mannitol for Mannitol salt agar, it does not provide an effective differential culture medium for the bacteria. To further
establish the reliability and the applicability of the results, it is recommended that the chemical analysis must be conducted
in order to determine specific component responsible for bacterial growth. Also, for the improvement of this study,
researchers recommend the use of an enhancer and a more advanced technology that could provide a much accurate and
specific result.
Keywords: Cauliflower, Staphylococcus aureus, bacterial growth, MSA, differential media

INTRODUCTION and lastly the Sodium Chloride which is 7.5%

Culture media contains nutrients and physical growth
parameters necessary for microbial growth. It could be a
solid or liquid preparation for the growth, transport and
storage of microorganism. It is important for
most microbiological tests to obtain pure cultures, to
grow and count microbial cells, and to cultivate and
select microorganisms. Differential culture medium
contain compounds that allow groups of microorganisms
to be visually distinguished by the appearance of the
colony or the surrounding media. Selective media
contain ingredients that inhibit the growth of some
organisms but allow others to grow (Acharya, 2010).
Mannitol Salt Agar (MSA) as a selective, differential and
indicator medium used to isolate and
identify Staphylococcus aureus from the clinical
specimen. Its purpose is to see if the microbe can
ferment the carbohydrate mannitol as a carbon source.
This is a medium that is composed of Peptone, as the
source of Nitrogen, Vitamin and Carbon, Phenol Red, as
the indicator for organisms that can ferment Mannitol
which is the major and differential ingredients of MSA
concentration can inhibit the other organism that cannot
tolerate the high saline levels (Acharya, 2013).
Staphylococcus aureus is a gram-positive bacterium that
is found on the skin and in the nasal passages of about a
quarter of humans. It is a facultative
anaerobe, meaning it can produce energy either in the
presence 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 a Laboratory Testing, this bacteria is isolated in
the mannitol salt agar, with a presence of Growth and
Yellow Medium that indicates S. aureus a Mannitol
Fermenter (Al-Araji, 2015).
Brassica oleracea var. botrytis also known as Cauliflower
is a flower 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
contain a variety of components, such as
polysaccharides, proteins, some lectins and polyphenols
like mannitol (Scott, 2016).This research study aims to

1 | Cauliflower as a Substitute Ingredient for Mannitol in Mannitol Salt Agar

 [WESLEYAN UNIVERSITY – PHILIPPINES
COLLEGE OF NURSING AND ALLIED MEDICAL SCIENCES
DEPARTMENT OF NURSING] May 23, 2018
produce a differential culture medium from the Brassica
oleracea var. botrytis for the growth of Staphylococcus
aureus.
METHODOLOGY
The design used in this study is Quasi-experimental
method, a non-randomized study. In the experiment only
the post test is measured.
Preparation of cauliflower powder
The researchers used a 10kg of Whole Cauliflower, it was
washed and 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 airtight
container for the next procedure.
Preparation of the culture media
The researchers used sterilized laboratory glasswares by
autoclaving at 121 Celsius for 15 minutes at 15 psi. The
100 grams powdered Cauliflower was mixed with 15
grams of Agar, .25 grams of Phenol Red 75 grams of NaCl
and 1000ml distilled water. The prepared agar mix was
gently heated until it dissolved and then autoclaved.
Preparation of mannitol salt agar
111 grams of Mannitol Salt Agar was dissolved in 1000
mL distilled water. Gently heated to completely dissolve
the medium for 5 minutes. Then, sterilized by
autoclaving at 121 Celsius for 15 minutes at 15 psi.
Preparation of agar plates
The prepared culture medium were extracted on the
disposable petri dish and allowed to stand for a minute,
then it was refrigerated at 4°C. Researchers did a
triplicate sample of each Agar Plate.
Streaking of bacteria and Incubation
The pure culture of Staphylococcus aureus and
Staphylococcus epidermidis were inoculated from the
Nutrient Agar. Using the overlapping streak plate
method, the bacteria from the pure culture was streaked
to the prepared Cauliflower agar and Mannitol Salt Agar
using a disposable inoculating loop. It is then, incubated
at an optimum temperature of 37°C and observed for
12th and 24th hour of incubation.
Biochemical testing
The incubated agar plates undergo biochemical testing
to identify the species that grow on the agar plates.
Researchers first describe the morphology on the agar
plate then perform the Gram Staining to identify if it is a
Gram Positive Bacteria and BBL Crystal Identification, for
2 | Cauliflower as a Substitute Ingredient for Mannitol in Mannitol Salt Agar
identification of the Staphylococcus aureus and
Staphylococcus epidermidis.
RESULTS
Mannitol Salt Agar and Staphylococcus aureus
The result of trial under the controlled conditions after
the 12th and 24thhours of observation. Staphylococcus
aureus shows an increase in growth through time in the
Mannitol Salt Agar and a change in color from red to
yellow.
Mannitol Salt Agar and Staphylococcus epidermidis
The result of the trial under the controlled conditions
after the 12th and 24thhours of observation. This shows
that Staphylococcus epidermidis shows an increase in
growth through time but no change in color at all in
Mannitol Salt Agar.
Cauliflower Agar and Staphylococcus aureus
The result of the trial under the controlled conditions
after the 12th and 24th of observation. It shows that
Staphylococcus aureus has the same growth through
time and it does not show any color changes at 12th and
24th hours.
Cauliflower Agar and Staphylococcus epidermidis
The result of the trial under the controlled conditions
after the 12th and 24th of observation. It shows that
Staphylococcus aureus has the same growth through
time and it does not show any color changes at 12th and
24th hours.
Morphology
It Shows the Morphology observed at 24th hour
incubation of Cauliflower agar for the identification of
the colonies. The colonies that grow on the agar are 1.0-
1.5 diameter, round and yellow which describe the
Staphylococcus aureus while on the other plate, it shows
0.8-1.2 mm diameter, round and white colonies which
describe Staphylococcus epidermidis.
Gram Stain
The result of the Gram Stain bacteria from the
Cauliflower Agar Plate are violet, this shows that the
organism on the plate are a Gram positive Bacteria which
includes the Staphylococcus aureus and Staphylococcus
epidermidis.
BBL Crystal Identification Kit
Using the BBL Crystal Identification it is confirmed that
the cultured bacteria from the Cauliflower agar are
Staphylococcus aureus and Staphylococcus epidermidis
when encoded on the computer.
 [WESLEYAN UNIVERSITY – PHILIPPINES
COLLEGE OF NURSING AND ALLIED MEDICAL SCIENCES
DEPARTMENT OF NURSING] May 23, 2018

CONCLUSIONS Briones J.S.M., Castro R.P.M., Ellazar J.M., Liwag

The Staphylococcus aureus grows in the Cauliflower Agar M.P.A.D., & Mangulabnan G.H.(2015). Growth of
which means it is able to provide enough nutrient for the Staphylococcus aureus in Blood Agar Plate Containing
both bacteria for it to grow but it is not an effective Different Anticoagulatant. Accesed from Wesleyan
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Staphylococcus epidermidis. This might because there is Bush,L.M. (2017).. Staphylococcus aureus Infections
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COLLEGE OF NURSING AND ALLIED MEDICAL SCIENCES
DEPARTMENT OF NURSING] May 23, 2018

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