Name: AGUSTIN, DANIELLA MIKAH E.
Section: BSN 1D
Group: 2
Virtual Laboratory Activity No. 1
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Agar We Go: Exploring Selective & Differential Media
I. OBJECTIVES
1. To differentiate bacteria based on their growth characteristics on selective and differential
media.
2. To understand the principle behind different culture media.
II. MATERIALS:
A. Laboratory Equipment
• Bunsen Burner
• Inoculating loop
B. Culture Media
• Mannitol salt agar • Hektoen Enteric Agar
• MacConkey’s agar • Blood Agar
• Eosin Methylene Blue Agar • Chocolate Agar
• Phenylethyl Alcohol Agar
C. Test Organisms
• Staphylococcus aureus • Streptococcus salivarius
• Staphylococcus epidermidis • Haemophilus influenza
• Escherichia coli
• Pseudomonas aeruginosa
• Enterobacter aerogenes
• Proteus vulgaris
• Shigella flexneri
• Salmonella typhimurium
• Streptococcus pyogenes
• Streptococcus pneumoniae
III. PROCEDURE
Preparation and Inoculation
• Prepare sterile agar plates → Label plates → Sterilize inoculating loop →
A. MSA:
S. aureus + S. epidermidis → streak → incubate
B. MacConkey Agar:
E. coli (left) + P. aeruginosa (right) → streak → incubate
C. EMB Agar:
E. coli + Enterobacter aerogenes → streak → incubate
D. PEA Agar:
S. aureus + S. epidermidis + Proteus vulgaris → streak → incubate
E. HE Agar:
Shigella flexneri + Salmonella typhimurium → streak → incubate
F. Blood Agar:
S. pyogenes + S. pneumoniae + S. salivarius → streak → incubate
G. Chocolate Agar:
Haemophilus influenzae → streak → incubate
• → Incubate all plates at 37°C for 24 hours
IV. DATA AND OBSERVATION
A. Mannitol Salt Agar (MSA) MacConkey’s Agar (MAC)
Figure 1. Culture of Staphylococcus aureus Figure 2. Culture of Staphylococcus
on Mannitol Salt Agar epidermis on Mannitol Salt Agar
B. MacConkey’s Agar (MAC
Figure 3. MacConkey’s agar with lactose
fermenters (left) and
non-lactose fermenters (right)
C. Eosin Methylene Blue (EMB) Agar
Figure 4. Escherichia coli on Figure 5. Enterobacter aerogenes Figure 6. Proteus
EMB agar on EMB agar vulgaris on EMB agar
D. Phenylethyl Alcohol (PEA) Agar
Figure 7. PEA agar plate with selective growth
of Gram-positive bacteria
E. Hektoen Enteric (HE) Agar
Figure 8. Growth of and Shigella flexneri and Salmonella typhimurium on Hektoen Enteric Agar
F. Blood Agar (Hemolysis Patterns)
Figure 9. Beta hemolysis Figure 10. Alpha hemolysis Figure 11. Gamma hemolysis
G. Chocolate Agar
Figure 12. Haemophilus influenzae on Chocolate Agar Plate
V. ANALYSIS/DISCUSSION
Selective and differential media effectively distinguished bacterial species based on growth and
biochemical reactions. In MSA, Staphylococcus aureus fermented mannitol (yellow), while
Staphylococcus epidermidis did not. In MacConkey’s agar, Escherichia coli appeared pink
(lactose fermenter) and Pseudomonas aeruginosa remained colorless. EMB agar showed E. coli
with a green metallic sheen, while Enterobacter aerogenes formed mucoid colonies.
PEA agar supported Gram-positive growth, while HE agar differentiated Salmonella typhimurium
(black centers) from Shigella flexneri (green colonies). Blood agar showed beta (Streptococcus
pyogenes), alpha (Streptococcus pneumoniae), and gamma (Streptococcus salivarius)
hemolysis. Chocolate agar supported Haemophilus influenzae. Results matched expected
characteristics
VI. EVALUATION
Possible errors include improper streaking, contamination, or incorrect loop handling. Variations
in incubation conditions may also affect results. Despite these, outcomes were generally accurate
and consistent. (May happen in actual laboratory setting.)
VII. CONCLUSION
Selective and differential media are essential for identifying bacteria based on growth and
biochemical properties. The experiment confirmed their role in distinguishing organisms and
supporting accurate microbial identification.
VIII. REFERENCES
Morello, Mizer, Wilson, and Granato. Microbiology in Patient Care, 6th Edition, 1998.
Madigan, M. T., Bender, K. S., Buckley, D. H., Sattley, W. M., & Stahl, D. A. (2021). Brock biology
of microorganisms (16th ed.). Pearson.
Tortora, G. J., Funke, B. R., & Case, C. L. (2019). Microbiology: An introduction (13th ed.).
Pearson.