Download Where Does the Analytical Testing Begin?

Wastewater Treatment
Plants & Bacteria
Strategies for Compliance
Best Practices for Effluent
Sampling
TANNY BUSBY
&
LAURA BONJONIA
ENVIRODYNE LABORATORIES, INC.
April 26, 2011
Where Does the Analytical Testing
Begin?
 Container
Preparation
 Sample Collection
 Chain of Custody
 Preservatives
 Field QC
What is a Representative
Sample?
Definition
Every Member of the Population has an
“Equal Chance” of being Selected
Typically a Population is very large
A Sample is a Subset of the Population
The sample reflect characteristics
of the Effluent
Identification of Sampling Points
 Effluents
 Influents
 Process
Control
 Collection Systems
DEFENSIBLE DATA
Sample Collection

Grab








e-Coli
Fecal Coliform
Oil & Grease
*DO
*pH
Cyanide
VOC
*Chlorine Residual

Composite








Metals
BNA
Pesticides
TPH
Minerals
Nutrients (Nitrogen)
BOD
TSS (Solids)
DEFENSIBLE DATA
QUALITY CONTROL
 Calibrate
Field Equipment - (Document in
field log) ~ pH, DO & Cl2 Meters
 Field Blanks
 Trip Blanks
 Rinseates
 Field and Lab Duplicates
 Field and Lab Matrix Spikes (MS)
 Laboratory control samples (LCS)
 Method Numbers
WHAT ARE COLIFORM

Gram stain
• Gram negative (turns
red)



Do not produce
spores
Bacilli (rod shaped)
Aerobic (require
oxygen) or
Facultative
Anaerobic
WHERE ARE COLIFORM(s)
FOUND?
 Surface
Water
 Decaying Plants
 Soil
 Intestinal Tract of Warm-blooded Animals
TYPES OF COLIFORM
BACTERIA
Total Coliform
E. coli
e.g. Salmonella (Typhoid Fever)
Shigella
Pseudomonas
Fecal Coliform
Enterobacter aerogenes
Free in the environment
e.g. soil & plants
Fecal streptococcus
Clostridium
~ Microbes ~
Commonly Regulated in
Wastewater Effluents
 Fecal



 E.


Coliform
200 CFU/100ml ~ Monthly Geo-Mean (Avg.)
400 CFU/100ml ~ 7 Day Geo-Mean (Avg.)
800 CFU/100ml ~ Single Grab Max.
coli ~ (Escherichia coli)
126 CFU/100ml ~ Monthly Geo-Mean (Avg.)
394 CFU/100ml ~ Single Grab Max.
 Enterococci

36 CFU/100ml ~ Monthly Geo-Mean (Avg.)
COLIFORM LAB TESTING
METHODS
-
Multiple Tube Fermentation (Quantifies)
- Reported as MPN (most probable number)
-
Membrane Filtration (Quantifies)
- Reported as CFU’s/100 mL (colony forming units)
-
Quanti-tray (Quantifies)
- Colilert – 18-24 hours
- Reported as MPN
- Positive sample turns yellow; E.coli fluorescence
-
Presence/Absence (Qualifies)
- Colilert, Colisure
- Positive sample turns yellow (Colilert); turns magenta (Colisure)
- E.coli fluorescence
E. coli

Membrane filtration

m-ColiBlue 24
• Total coliform and
E. coli simultaneously
• E. coli – Blue
colonies
• Total coliform – Red
or blue colonies

Other methods are
approved
ENTEROCOCCI PLATE

Membrane filtration




SM 9230 C
CFUs/100 ml
Blue colonies
Other methods

Enterolert
• MPN (Most Probable
Number)
COLILERT- QUANTITRAY
Incubate 18-24 hours- Most Probable Number
TOTAL
COLIFORMS
E.Coli
(UV Light)
SAMPLING HOLD TIMES
 Fecal
Coliform, E. coli and Enterococci
 Grab (Not Composite)
 Sterile container
 Hold Time ~ Not to Exceed a Total of 8Hrs

6 hours transport + 2 hours set up
Micro Samples
How to Collect Effluent
 Directly
into Sterile Testing Container
How to Collect and Split Samples
Total Coliform Qualitative Analysis
(PRESENCE~ABSENCE)
COLILERT-18
-Incubate 18-22 hours
COLILERT
-Incubate 24-28 hours
COLISURE
-Incubate 24-48 hours
E.coli
positive
(UV Light)