Bacillus cereus is a Gram-positive, rod-shaped, aerobic, motile, beta hemolytic bacterium commonly found in soil and food.

Reference:

• Been, M. De. “Comparative analysis of two-component signal transduction systems of Bacillus cereus, Bacillus thuringiensis and Bacillus anthracis.” Microbiology 152.10 (2006): 3035-048. Web.

Date collected: February 8, 2017

Methods for isolation and identification:

1. Collection and Sampling of Environmental Samples at Environmental Education Center (Figure 1)
2. Selection of bacteria from plate to be analyzed (Figure 2)
3. Polymerase Chain Reaction to amplify 16S rRNA
4. Restriction Enzyme Digestion & Gel Electrophoresis (Figure 3)
5. DNA Sequencing Analysis/Identification

Figure 1. Sampling site locations in Lancer Park. The top site is Buffalo Creek and the lower site is the pond where samples were taken

Figure 2: picture of Bacillus cereus

Results:

• MspI digestion (Figure 3): A 1,500 bp product was amplified by PCR. Upon digestion with a band at 500 bp digested and a 1000 bp undigested.

Figure 3: Mspi digestion of Bacillus cereus PCR product

• Sequence analysis (Figure 4): The sequenced PCR product generated 887 bases. NCBI BLAST analysis revealed 99% identity with 887 of the bases matching the 16s rRNA gene of Bacillus cereus (Figure 4).

• Figure 4: The BLAST sequence match of the sample bacteria to Bacillus cereus

Contributed by: Matthew W. Bowman & Carly Carter, BIOL 250 – Spring 2017, Longwood University