Results Information
28044 James Dr.
Warren, MI
48092
Phone: 586-806-4243
Fax: 313- 577-8616
Email: info@helixbiolab.com
Customer ID: Huron River Watershed
Project #: P-173A
Sample Date Received: 6/5/12
Date of Analysis: 6/6/12 – 6/12/12
Sample Type: Water
Analysis Type: MST
Procedure:
Submitted water samples from Honey Creek were tested for the presence of source
specific Bacteroides bacteria using target DNA markers. Samples received were
from June 5, 2012 collection.
Submitted water samples had the following sample designation:
HC1
HC4
HC7
HC12
HC14
HC15
HC17
Samples HC1, HC4, HC7 and HC17 were analyzed using MST
100 ml of each water sample tested was suctioned filtered through a 0.45 micron
filter membrane that proceeded to capture any bacteria present in the water
samples. Each individual filter membrane containing captured bacteria was
removed from each individual filter column following filtration and used to grow a
pre-enrichment bacterial culture on media selective for Bacteroides at 37 degrees C
for 72 hours. After incubation, bacterial DNA was extracted from the filter
membranes used in the pre-enrichment cultures.
Bacterial DNA extractions were then used as DNA template for qualitative PCR
analysis. PCR analysis incorporated oligonucleotide primers that target specific
marker DNA sequences unique to source specific Bacteroides bacteria.
Appropriate positive and negative control samples were included in PCR
experimentation. Following the PCR procedure, the PCR products were run on a
1.5% agarose gel at 90V for 60 minutes, after which the agarose gel containing the
PCR products was visualized under UV light and examined for PCR amplification
products that represent the amplification of the target DNA markers.
Replicates of each sample were run for quality assurance purposes and to confirm
the observed results.
Results
Results of the qualitative PCR testing indicate the presence (positive) or absence
(negative) of source specific DNA markers through PCR amplification of source
specific DNA marker sequences.
A sample testing positive for a source specific DNA marker indicates that the
marker DNA sequence representing the specific bacteria was amplified, thus
confirming the absence of that source specific DNA marker in the sample.
Sample
Source DNA Marker
Result
HC1
HC1
HC1
HC1
HC1
HC1
General Bacteroides
Human
Horse
Dog
Goose
Cow
+
+
-
HC4
HC4
HC4
HC4
HC4
HC4
General Bacteroides
Human
Horse
Dog
Goose
Cow
+
+
+
-
HC7
HC7
HC7
HC7
HC7
HC7
General Bacteroides
Human
Horse
Dog
Goose
Cow
+
+
-
HC17
HC17
HC17
HC17
HC17
HC17
General Bacteroides
Human
Horse
Dog
Goose
Cow
+
+
-
A positive result for a source specific DNA marker at a collection station implies
that source is contributing to fecal contamination that may have been determined at
the collection station during the collection period. A negative result for a source
specific DNA marker at a collection station implies that source is not contributing
to fecal contamination that may have been determined at the at the collection
station during the collection period.
These results were verified by Tom Prychitko, Laboratory Director for Helix
Biological Laboratory.