Manzano, Joe Anthony Nicolas, Ellha Mae Nicole Park, Sang Keon 11/18/19
Medalla, Milen Angelie Pambid, Gyle Group 8 Asst. Prof. De Jesus & Asst Prof. Medina
IV. GUIDE QUESTIONS
A.
1. Classification of Biochemical Tests Reactions Based on the ff:
Presence of Presence of Carbohydrate Protein Catabolism
Extracellular Intracellular Fermentation
Enzymes Enzymes
Starch Hydrolysis IMViC Methyl Red Test Protein Hydrolysis
Test (Indole,Methyl Red, and Voges
Voges Proskauer, Proskauer Test
and Citrate (Organic Acid
Utilization) Test Fermentation)
Gelatin Hydrolysis Triple Sugar Iron Mannitol Salt Agar
Test Test Test (Mannitol
Fermentation)
Urease Test Nitrate Reduction MacConkey Agar
Test Test
(Lactose
Fermentation)
Urease Test Triple Sugar Iron
Test
(Carbohydrate
Utilization Test)
Catalase Test Motility Indole
Ornithine
Decarboxylation
Test
(Dextrose
Fermentation)
Oxidase Test
2. Other Tests that can be used in the identification of unknown.
a. Coagulase Test
This test is for the detection of coagulase enzyme produced by Staphylococcus aureus,
having a lyophilic rabbit plasma containing EDTA (Ethylenediaminetetraacetic Acid) to
constitute the test. Specifically, it is used to differentiate Staphylococcus aureus (positive)
from Coagulase Negative Staphylococcus (CONS).
b. Camp Test
aving to rarely give
This test is effective for the identification of Streptococcus agalactiae, h
false positives with other Streptococcus. This test is merely used for the presumption of
Group B beta-hemolytic streptococci, Streptococcus agalactiae for its identification.
c. Spirit Blue Agar Test
This test is for the detection for lipolytic (lipase-producing) microorganisms, having the spirit
blue to be the indicator of lipolysis, and the agar as the solidifying agent. This test will result
of lipolytic microorganisms to metabolize the lipid inside the medium, forming colonies with
halos that indicates lipolysis.
d. Bile Solubility Test
The bile solubility test is based on the observation of Streptococcus pneumoniae t o be
visibly lyse when such 2% or 10% of sodium desoxycholate (bile salts) is applied to the
ot to lyse. Specifically,
colony under such time and temperature, having other Streptococci n
this test is to be used only for the differentiation of S. pneumoniae from other
alpha-hemolytic streptococci; having this test to not be reliable when used with other old
cultures, since they have no more active enzymes, resulting in a negative result.
e. Potassium Hydroxide Test
This highly corrosive chemical is a test for the detection of fungal elements present in a
cultured media in either general purpose fungal culture media, such as Sabouraud Dextrose
Agar (SDA) or a specific media based on the type of anticipated isolate.
3. Many culture media produce colorful solution due to the presence of which are water soluble
organic compounds that change color whenever there is a change in pH. Enumerate the
indicators used, indicate the tests where they are used and give their specific reactions.
a. Phenol Red
i. Urease Test: from yellow to pink in color.
ii. MSA Test: from red to yellow in color.
iii. TSI Test: medium will turn yellow in color, if it can ferment three sugar present
b. Neutral Red
i. MacConkey Agar Test: from red to bright pink; color change means the
fermentation is acidic in pH.
c. Bromothymol Blue
i. Simmon’s Citrate Agar: from green to blue in color; caused by the alkaline pH.
d. Bromocresol Purple
i. MIO Test: from purple to yellow in color.
e. Methyl Red
i. MRVP Test: pH is below 4.4 if there is a red color change, pH is above 6.0 if there is
a yellow color change.
f. Iodine Solution
i. Starch Hydrolysis: medium will turn dark blue to brown in color.
B.
1. An undercooked boiled egg was received in the microbiology laboratory with request culture
and identification. This egg was served in a pack lunch during a birthday party. All guests
who ate this egg were brought to the hospital for vomiting and dehydration. Some had bouts
of diarrhea. The signs and symptoms were short-lived after treatment was received. The egg
was homogenized in tryptic soy broth. This was plated in sheep’s blood agar plate and
macconkey agar plate. After 24 hours of incubation, the following observations were noted:
● Sheep’s Blood agar plate: 4+ (many) beta creamy white
● MacConkey Agar Plate: no growth
● The gram stain result of the isolate yielded gram positive cocci, mostly in clusters.
If Staphylococcus aureus is suspected what is the course of action to identify this bacteria.
Staphylococcus aureus i s a gram-positive bacteria. MacConkey agar Since it is known in the gram
stain result that the isolated organism is a gram positive cocci, catalase test can be used to know whether
ositive result will give us
this gram positive cocci belongs to streptococcus or staphylococcus, p
staphylococci organism identity, it is then followed by coagulase/protein A test to distinguish whether the
isolated organism is S. aureus or belong to a different strain of Staphylococcus organism, positive result will
tell us that the isolated organism is Staphylococcus aureus. The process mentioned above directly tell us
whether the isolated gram positive cocci belongs to Staphylococcus aureus or not. We can also run a
different set procedure by doing the catalase test first, again Staphylococcus aureus have a positive result
in this test. TSA deep test is then followed, Staphylococcus aureus and other strain of Staphylococcus
belongs to the facultative anaerobic. In order to separate the Staphylococcus aureus to other strains of
Staphylococcus, we can inoculate the unknown organism to Mannitol Salt Agar, if it yields a positive result,
the unknown organism is Staphylococcus aureus.
REFERENCES
Baird-Parker, A. C. (1963). A classification of micrococci and staphylococci based on physiological and
biochemical tests. Microbiology, 30(3), 409-427.
Bascomb S, Manafi M. Use of enzyme tests in characterization and identification of aerobic and
facultatively anaerobic gram-positive cocci. Clin Microbiol Rev. 1998;11(2):318–340.
Dhanya, Ari, Lyn, & Riket. (2019, August 26). KOH Preparation Test: Principle, Procedure, Results and
Uses . Retrieved from
[Link]
Jutte, P., Lazzeri, E., Sconfienza, L. M., Cassar-Pullicino, V., Trampuz, A., Petrosillo, N., & Signore, A.
(2014). Diagnostic flowcharts in osteomyelitis, spondylodiscitis and prosthetic joint infection. THE
QUARTERLY JOURNAL OF NUCLEAR MEDICINE AND MOLECULAR IMAGING.
Kelvin, R., Brian, Arora, A., Dr.R, & Samsuddin. (2019, January 10). CAMP Test: Principle, Procedure and
results. Retrieved from [Link]
Lowy, F. D. (1998). Staphylococcus aureus infections. New England journal of medicine, 339(8), 520-532.
Nirmal, Kathy, Gull, Sukhadia, H., Kavitha, & Khan, P. (2019, April 30). Coagulase Test: Principle,
procedure and interpretation. Retrieved from
[Link]
Perry, J. D., Rennison, C., Butterworth, L. A., Hopley, A. L., & Gould, F. K. (2003). Evaluation of S. aureus
ID, a new chromogenic agar medium for detection of Staphylococcus aureus. Journal of clinical
microbiology, 41(12), 5695-5698.
Pokhrel, P. (2015, July 19). Bile Solubility Test- Principle, Procedure, Result Interpretation, Examples and
Limitation. Retrieved from
[Link]
nd-limitation/.
Starr, M. P. (1941, April 4). SPIRIT BLUE AGAR: A MEDIUM FOR THE DETECTION OF LIPOLYTIC
MICROORGANISMS. Retrieved from [Link]