EPA Method 1106.1

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EPA Method 1106.1:
Enterococci in water by MF using mE-EI Agar. Official Name: Enterococci in water by membrane filtration using membrane-Enterococcus-Esculin Iron Agar (mE-EIA)

Summary:
This method provides a direct count of bacteria in water based on the development of colonies on the surface of the membrane filter. A water sample is filtered through the membrane which retains the bacteria. Following filtration, the membrane is placed on a selective medium, mE agar, and incubated at 41oC + 0.5oC for 48 + 3 hours. The filter is then transferred to a differential medium, EIA, and incubated at 41oC + 0.5oC for 20-30 minutes. Pink to red enterococci colonies on mE will develop a black or reddish-brown precipitate on the underside of the filter after transfer to EIA. A fluorescent lamp with a magnifying lens is used for counting to give maximum visibility of colonies.

Scope:
This method describes a membrane filter procedure for the detection and enumeration of the enterococci bacteria in ambient water. The enterococci test is recommended as a measure of ambient fresh and marine recreation water quality.

Citation:
USEPA, 2006, Method 1106.1: Enterococci in water by membrane filtration using membrane-Enterococcus-Esculin Iron Agar (mE-EIA): U.S. Environmental Protection Agency Report 821-R-06-008, 42 p.

Interferences:
Water samples containing colloidal or suspended particulate materials can clog the membrane filter and prevent filtration, or cause spreading of bacterial colonies which could interfere with enumeration and identification of target colonies.

QC Requirements:
The minimum analytical QC requirements for the analysis of samples using this method include routine analysis of positive and negative controls, filter sterility checks, method blanks, and media sterility checks.Additional analytical QC for the analysis of samples using this method include an initial demonstration of laboratory capability through performance of the initial precision and recovery (IPR) analyses, and ongoing demonstration of laboratory capability through performance of the ongoing precision and recovery (OPR) analysis. For the IPR and OPR analyses, it is necessary to spike PBS samples with either laboratory-prepared spiking suspensions or BioBalls.

Maximum Holding Time:
Analysis preferably within 2 hours of collection; max. transport time to lab is 6 hours, and samples should be processed within 2 hours of receipt at lab.

Media:
WATER

Subcategory:
Microbiological

Concentration:
20-80 colonies per membrane

Precision:
The precision was evaluated in both fresh and marine recreational waters and in sewage treatment plant effluents.

Detection:
If counts from all membranes are zero, calculate using count from largest filtration volume. Calculate the number of colonies per 100 mL that would have been reported if there had been one colony on the filter representing the largest filtration volume. See the method report for examples on calculating the colony counts.Report as being "less than" the calculated number.

Revision Number:
6-Jul