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Emerging Diagnostics - A Sneak Peak at How Tomorrow’s Technologies are Being Used at the University of Wisconsin Today. Sharon C. Long. Today’s Objective. To get you thinking about new tools and approaches to diagnosing the biology in your processes Activated sludge Biosolids Source water

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Emerging Diagnostics - A Sneak Peak at How Tomorrow’s Technologies are Being Used at the University of Wisconsin Today

Sharon C. Long


Today s objective
Today’s Objective Technologies are Being Used at the University of Wisconsin Today

To get you thinking about new tools and approaches to diagnosing the biology in your processes

  • Activated sludge

  • Biosolids

  • Source water

  • Distribution systems

  • Etc.


Omics
“Omics” Technologies are Being Used at the University of Wisconsin Today

  • Metabolomics – aims to compare differences between biological samples based on their metabolic profiles

  • Genomics – using sequences of DNA and RNA to develop useful biological knowledge

  • Proteomics –used to relate microbial activities to the identity of organisms in multispecies communities


Today s examples
Today’s Examples Technologies are Being Used at the University of Wisconsin Today

  • Using ATP profiles to evaluate overall microbial community changes

  • Recognition of short sequences of DNA to evaluate presence of certain target microorganism - FISH

  • Measuring the presence of short sequences of DNA to evaluate presence of certain target microorganism - PCR

  • Sequencing the DNA of environmental samples to assess community structure


ATP Technologies are Being Used at the University of Wisconsin Today

  • Transports chemical energy within cells for metabolism

  • Present in all living cells

  • Amount in a sample is roughly proportional to the number of cells present

  • Measure the amount of light produced using firefly enzymes


Study of microbial occurrence in wells
Study of Microbial Occurrence Technologies are Being Used at the University of Wisconsin Today in Wells

370

ME/ml

45,300

ME/ml

12,600

ME/ml

860

ME/ml

460

ME/ml

1,912,000

ME/ml

801,000

ME/ml

241,000

ME/ml

8,620

ME/ml

5,200

ME/ml

From: Andrew Jacque, PhD, PE, Water Quality Investigations LLC and Town & Country Engineering


Well study

370 Technologies are Being Used at the University of Wisconsin Today

ME/ml

Well Study

45,300

ME/ml

12,600

ME/ml

860

ME/ml

From: Andrew Jacque, PhD, PE, Water Quality Investigations LLC and Town & Country Engineering


Fluorescent in situ hybridization fish
Fluorescent Technologies are Being Used at the University of Wisconsin Today in situ Hybridization (FISH)

  • Use short sequences of single stranded DNA complimentary to a sequence unique or general to the target organism(s)

  • Attach a fluorescent dye

  • Prepare a sample

    • Permeation

    • Hybridization

  • Quantify using microscopy


Applications of fish
Applications of FISH Technologies are Being Used at the University of Wisconsin Today

  • Diagnosis of foaming in biosolids anaerobic digesters

Klare Keadle, Zachary Carroll, Daniel R. Noguera, and Sharon C. Long


Foaming and fish results
Foaming and FISH Results Technologies are Being Used at the University of Wisconsin Today

  • Provided qualitative confirmation of the presence and relative density of target filaments

  • M. parvicella was confirmed to be numerous

  • Other known filamentous Archeae also present in digesters


Applications of fish 2
Applications of FISH 2 Technologies are Being Used at the University of Wisconsin Today

  • Characterize microorganisms in aerated anoxic Enhanced Biological Phosphorus Removal

Ramesh K. Goel, Patricia Sanhueza and Daniel R. Noguera


Ebpr fish results

120 Technologies are Being Used at the University of Wisconsin Today

Run II ( 0.3 % O

)

2

100

80

60

120

%

Run III ( 0.6 % O

)

2

40

100

% P removal

20

% Rhodocyclus related

80

120

Run IV ( 1.2 % O

)

2

0

0

10

20

30

40

50

60

100

60

Time (days)

%

80

40

% P removal

60

20

%

% Rhodocyclus related

40

0

% P removal

0

5

10

15

20

25

30

20

% Rhodocyclus related

Time (days)

0

0

10

20

30

40

Time (days)

EBPR FISH Results

More than 90 % Population was Rhodocyclus related


Polymerase chain reaction pcr
Polymerase Chain Reaction (PCR) Technologies are Being Used at the University of Wisconsin Today

  • Use of Taq polymerase

  • Target short (~100 to 400 bp), unique sequences of DNA or RNA for species identification

  • Target broadly occurring sequences of DNA for organism family identification

  • Products may also be sequenced to yield further information

Exponential Replication

1 2 3 4 5


Measure presence of stec in wastewater
Measure Presence of STEC in Wastewater Technologies are Being Used at the University of Wisconsin Today

(A) E. coli O157:H7

(B) Non-STEC E. coli or Shigella

(C) E. coli absent

(D) Toxigenic organism, not STEC, not Shigella

(E) Non-O157:H7 STEC or Shigella

India Mansour, Mark Walter, and Sharon C. Long


Dna sequencing
DNA Sequencing Technologies are Being Used at the University of Wisconsin Today

Trevor Ghylin


Database of wastewater microbes proprietary
Database of Technologies are Being Used at the University of Wisconsin Today Wastewater Microbes (Proprietary)


Ebpr genomic study
EBPR Genomic Study Technologies are Being Used at the University of Wisconsin Today


Acknowledgements
Acknowledgements Technologies are Being Used at the University of Wisconsin Today


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