Flavobacterium aquidurense is a dark orange, circular colony found in the water of the Appomattox River. It has been previously found in Northern Germany in a hard water creek (Cousin, S., 2007).

References:

1. Cousin, S., Pauker, O., and Stackebrandt, E. “Flavobacterium aquidurense sp. nov. and Flavobacterium hercynium sp. nov., from a hard-water creek.” Int. J. Syst. Evol. Microbiol. (2007) 57:243-249.

Date Collected: February 8, 2017

Methods for isolation and identification:

• A water sample was taken from on top the water of the Appomattox River (Figure 1). 100μl of the sample was placed on an agar plate and was incubated at 25 degrees Celsius for 48 hours. The plate was parafilmed and placed in the refrigerator for seven days.
• A dark orange, circular colony (Figure 2) was selected for 16S rRNA gene sequencing by PCR amplification.
• The PCR product was sent for DNA sequencing to identify the species of the bacteria.

Figure 1. Site of collection

Figure 2. Selected colony for identification

Results:

• MspI digestion (Figure 3): A bold band was produced at 2,000 bp by PCR.

Figure 3. Gel electrophoresis results of Appomattox River Water PCR product and + MSP1.

• DNA Sequence: The sequenced PCR product produced 810 quality base pairs that were used to identify the bacteria as Flavobacterium aquidurense. The chromatogram of the sequence is available as a pdf (HM03_PREMIX_JF7556_25). NCBI BLAST revealed 99% similarity with 4 gaps out of 810 base pairs (Figure 4).

Figure 4. NCBI BLAST results of chosen colony.

Contributed by: Megan Bland and Hannah Hatke, BIOL 250 Spring 2017