Proposed title: Sampling of Wastewater and Biosolids for
Bacteria and Viruses
Chapter no.:
_________
Author(s): John Scott Meschke
Department
of Environmental and Occupational Health Sciences
University of
Washington
4225 Roosevelt Way NE,
suite 2338
Seattle, WA 98105-6099
Notes:
·
Material
in this chapter may overlap with other planned chapters on Detection of
Pathogens in Sludges, etc. (Judy Blackbeard) and Assessing the Efficacy of
Wastewater Treatment (Wesley Pipes).
·
Content
covered in this chapter will include microbiological sampling considerations,
primary and secondary concentration techniques, and separation/purification processes
for target microbes in raw waste water, primary, secondary, and final effluent,
and biosolids.
·
Techniques
relevant to viruses, bacteria, and eukaryotic microbes will be covered.
Application to both pathogenic and non-pathogenic targets will be discussed.
·
Culture
and other detection techniques will not be covered in detail (rather they will
only be referred to as they related to the upstream sampling, concentration,
and purification techniques).
Proposed topics
1)
Introduction:
overview of wastewater treatment (purpose, processes, communities involved,
etc.), matrix description/characterization (raw wastewater, primary effluent,
secondary effluent, final/tertiary effluents, biosolids), etc.
2)
Relevant
sampling schemes: purpose/rationale for sampling, types of
sampling schemes and appropriate uses, relevant regulation, qualitative and quantitative
approaches, etc.
3)
Methods
for Sampling of Viruses
a.
Viral
Targets: pathogens and phage
b.
Raw
Wastewater; Primary, Secondary, Advanced, and Final Effluents
i. Sample Collection:
sample volumes, sample preservation, time to processing, etc.
ii. Concentration
Methods: filtration, precipitation, acid
adsorption/elution and centrifugal methods
iii. Separation/Purification
Techniques: phase separation, organic extraction, serial filtration, gradient
centrifugation, etc.
c.
Biosolids
i. Sample
Collection: sample mass, preservation, time to processing, etc.
ii. Elution/Extraction
Techniques
iii. Separation/Purification
Techniques: phase separation, organic extraction, serial filtration, gradient
centrifugation, etc.
d.
Nucleic
Acid Purification
4)
Methods
for Sampling of Bacteria
a.
Bacterial
Targets: indicator bacteria, pathogens, process bacteria (active and nuisance)
b.
Raw
Wastewater; Primary, Secondary, Advanced, and Final Effluents
i. Concentration and
Enrichment Methods
ii. Separation Methods
c.
Biosolids
i. Elution/Enrichment
Methods
ii. Separation
Methods
d.
Nucleic
Acid Purification
5)
Methods
for Sampling of Eukaryotic Microbes
a.
Eukaryotic
Targets: algae, fungi, protozoa, helminth ova
b.
Raw
Wastewater; Primary, Secondary, Advanced, and Final Effluents
i. Concentration
Methods
ii. Separation
Methods
c.
Biosolids
i. Elution Methods
ii. Separation
Methods
d.
Nucleic
Acid Purification
6)
Use
of Sampling and Recovery Controls
Chapter
Highlights
The
following concepts will be conveyed in this chapter:
1. Matrix effects of sampling methods
2. Impact of target on sampling methods
3. Sampling for
whole cell or culture based detection versus nucleic acid based detection.
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