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Sensorer til måling af mikrobiel vandkvalitet SEMIVA projektet – hvad ... · 1 Sensorer til...
Transcript of Sensorer til måling af mikrobiel vandkvalitet SEMIVA projektet – hvad ... · 1 Sensorer til...
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Sensorer til måling af mikrobiel vandkvalitet SEMIVA projektet – hvad findes og hvad er undervejs
Hans-Jørgen Albrechtsen
Professor, Head of Section - Urban Water Systems
DANVA
Drikkevandssensorer 2016 – udstilling
og vidensdeling
27. januar 2016,Vandhuset,
Skanderborg
SEMIVA SEnsorer til måling af MIkrobiel VAndkvalitet
HOFOR A/S (Ann-Katrin Pedersen)
Aarhus Vand A/S (Jørn-Ole Andreasen)
VandCenter Syd A/S (Anne Esbjørn)
Kalundborg Forsyning A/S (Pernille Ingildsen)
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’Den ideelle sensor’ Forsyningernes definition
Installeres online eller at-line
Detektere indikatororganismer (E. coli og total coliforme)
Have høj følsomhed til detektion af lave koncentrationer af mikroorganismer i drikkevand
Give hurtigt svar
Kræve lav vedligeholdelse
Give lave forekomst af falske-positive
Arbejdsmetode
Opsummering af erfaringer med sensorer til monitering af mikrobiel vandkvalitet fra danske vandforsyninger - Spørgeskema
Fokuseret internet søgning
Opgørelse af relevante forskningsprojekter
Dialog med sensorproducenter
Telefoninterviews med fagfolk
Gennemgang af den videnskabelige litteratur
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Teknologier
på markedet
under udvikling eller i dokumentationsfase
på forskningsniveau
Teknologier på markedet Påvisning af coliforme og E. coli
Enzymatisk
Samme princip som Colilert
E.coli β-galactosidase activity (yellow)
Coliforme β-glucuronidase activity (fluorescence)
P. aeruginosa substrate hydrolysation
Total bacteria alkaline phosphatase activity
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Påvisning af coliforme og E. coli
Technology name
ALARM CALM Coliguard
BACTcontrol) aquaBio TECTA ColiMinder
Company Colifast A/S Colifast A/S MicroLAN
(mbOnline) ADASA Veolia
VMW (Vienna Monitoring Solutions)
Country Norway Norway The Netherlands Spain Canada/
Switzerland Austria
Analysed parameter
TC or EC TC, EC, P.
Aeruginosa (two simultaneously)
TC or EC TC and EC
simultaneously TC and EC
simultaneously TC, EC, or total
bacteria
Application DW Raw water,
process water, wastewater
DW, bathing water, process
water
DW, wastewater, bathing water
DW, wastewater, reclaim water
Sewage, surface water, DW soon
tested
Validation ETV EU project
Demowatercoli Research paper Currently not ETV N/A
Measuring unit
+/- +/-, MPN, CFU pmol
MUF/min/100 mL MPN/100 mL
+/- estimation of bacteria level
MUF/100 mL
Detection limit
1 cfu/100 mL 1 CFU/mL dep. on measuring mode
0.1 pmol MUF/100 mL
1 cfu/100 mL 1 cfu/100 mL 0.8 mMFU/100 mL
Installation at-line at-line at-line at-line Manual sampling
loading/ autosampling
at-line
Response time*
6-15 h (max 1 sample/day)
4-12 h 2-4 h 3-18 h 2-18 h 10-30 min (only
tested for surface water)
Sampling volume
100 mL 25-100 mL 20-3000 mL N/A N/A 6.5 mL
Maintenance Reagent refill
every 20 tests, yearly service
Reagent refill weekly to
monthly. Service twice a year
Reagents refill every 1-3 months for 4 tests/day13
Every 15 d reagent
replacement & monthly cleaning
N/A N/A for drinking water – currently
being tested
Market 190,000 DKK More expensive
than ALARM 300,000 DKK or
rental
174,000 DKK + 15,000 DKK yr maintenance
135,000 DKK For rent
Comments
HOFOR A/S, Trondheim and
Stavanger utilities
Stavanger utility
Enterococci i 2016? Experience by HOFOR A/S and
Nordvand A/S
Did not reply to inquiry. Svendborg
Vand A/S, Nordvand A/S,
HOFOR A/S with manual version
Currently working on new version
that incorporates pc
Coliforme / E. coli – sensorer
Teknologier:
‘ALARM’, ‘CALM’, ‘Coliguard’, ‘aquaBio’, ‘TECTA’, ‘Coliminder’
Automatiserede (excl. TECTA)
Enzymbaseret
Inkuberingstid til vækst af bakterier (2-18 timer)
Konvertering af enzym aktivitet til bakterietal - udfording
Forbrug af reagenser (refill uge/måned)
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Coliminder, aquaBio
Ingen dansk erfaring
ALARM
HOFOR A/S: data every 24 hours
Disadvantage: not for both TC EC at the same time)
Presence/absence.
Operation is quite stable, without excessive maintenance needs
Trondheim utility, 200,000 people: 3-4 years. infiltrated surface water, raw water intake to monitor for contamination
Several manure or sewage contamination detected, correlated roughly with grab sampling, good contamination indicator
User friendly system, only requires a short training course
Planning one more ‘ALARM’ at the lake water intake point
Rogaland (Stavanger) 300,000 people: 2 ‘ALARM’ sensors
Surface water intake points and monitor for E. coli every 4 h
Contamination cases detected, correlated roughly with weekly grab samples experience
positive
Concern: high initial and analysis cost, planning to continue using them
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CALM
Rogaland (Stavanger) 300,000 people
2 ‘ALARM’ sensors and 2 ‘CALM’ sensors
Overall experience is positive
Only expressed concern: high initial and analysis cost, especially of the ‘CALM’ sensor
Planning to continue using them
Coliguard
Nordvand A/S and HOFOR A/S
measure E. coli and total coliforms
4 times a day, low detection limit.
high maintenance needs
consider it an expensive technology
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TECTA
Nordvand A/S, HOFOR A/S, Svendborg Vand A/S
manual version
rather positive, user-friendly
response time of 2-18 hours is shorter than the traditional culture based methods, yet giving accurate results
only partly quantitative
mainly used during holidays, when laboratory closed
Påvisning af coliforme og E. coli
Technology name
ALARM CALM Coliguard
BACTcontrol) aquaBio TECTA ColiMinder
Company Colifast A/S Colifast A/S MicroLAN
(mbOnline) ADASA Veolia
VMW (Vienna Monitoring Solutions)
Country Norway Norway The Netherlands Spain Canada/
Switzerland Austria
Analysed parameter
TC or EC TC, EC, P.
Aeruginosa (two simultaneously)
TC or EC TC and EC
simultaneously TC and EC
simultaneously TC, EC, or total
bacteria
Application DW Raw water,
process water, wastewater
DW, bathing water, process
water
DW, wastewater, bathing water
DW, wastewater, reclaim water
Sewage, surface water, DW soon
tested
Validation ETV EU project
Demowatercoli Research paper Currently not ETV N/A
Measuring unit
+/- +/-, MPN, CFU pmol
MUF/min/100 mL MPN/100 mL
+/- estimation of bacteria level
MUF/100 mL
Detection limit
1 cfu/100 mL 1 CFU/mL dep. on measuring mode
0.1 pmol MUF/100 mL
1 cfu/100 mL 1 cfu/100 mL 0.8 mMFU/100 mL
Installation at-line at-line at-line at-line Manual sampling
loading/ autosampling
at-line
Response time*
6-15 h (max 1 sample/day)
4-12 h 2-4 h 3-18 h 2-18 h 10-30 min (only
tested for surface water)
Sampling volume
100 mL 25-100 mL 20-3000 mL N/A N/A 6.5 mL
Maintenance Reagent refill
every 20 tests, yearly service
Reagent refill weekly to
monthly. Service twice a year
Reagents refill every 1-3 months for 4 tests/day13
Every 15 d reagent
replacement & monthly cleaning
N/A N/A for drinking water – currently
being tested
Market 190,000 DKK More expensive
than ALARM 300,000 DKK or
rental
174,000 DKK + 15,000 DKK yr maintenance
135,000 DKK For rent
Comments
HOFOR A/S, Trondheim and
Stavanger utilities
Stavanger utility
Enterococci i 2016? Experience by HOFOR A/S and
Nordvand A/S
Did not reply to inquiry. Svendborg
Vand A/S, Nordvand A/S,
HOFOR A/S with manual version
Currently working on new version
that incorporates pc
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Andre teknologier på markedet Technology name Aquascope
Desktop microscope
7000 RMS BACMON EZ-ATP Biocounter
Company Biotrack 4-deep Mettler Toledo A/S Grundfos A/S Applitek Biosensores
Country The Netherlands Canada International Denmark Israel,
Belgium Spain
Parameter Total bacteria, E. coli, Enterococci
or Legionella
Particles, bacteria or other
organisms
Bacteria and particles
(simultaneously)
Particles and total bacteria
ATP N/A
Principle Filter cytometry
& FISH Optical Optical Optical
Chemiluminescence
N/A
Application N/A Medical (cancer research, urine and blood tests)
Pharmaceutical waters
Drinking water Drinking
water and wastewater
Wastewater, beverage
Validation N/A N/A N/A MS in prep N/A N/A
Measuring unit N/A Particles or cells/mL
Bacteria and particles/mL
Cells and particles/mL
N/A N/A
Detection limit 1 cell/mL N/A for drinking
water 1 Biocount (no
conversion) 160 cells/mL N/A 103 cells/ mL
Installation N/A + flow cell at-line at-line at-line at-line
Response time 20-45 min Immediate N/A 10 min N/A 15 min
Sampling volume 1-500 mL N/A N/A 6 µL fixed in the flow cell
N/A N/A
Maintenance N/A Every day wiping
glass and flushing flow cell
N/A Regular change
of flow N/A N/A
Market N/A
Rental, 200,000 DKK + min.
40,000 DKK for training
N/A For rent N/A N/A
Comments No reply Microscope
needs operator at least remotely
US technology 'RMS-ON90' just
acquired Information not
available
Experience TREFOR VAND A/S, Aarhus Vand A/S,
HOFOR A/S
No reply No reply
Aquascope
Company: Biotrack
Country The Netherlands
Parameter Total bacteria, E. coli, Enterococci or Legionella
Principle Filter cytometry & FISH (Fluorescence in situ hybridization)
Detection limit 1 cell/mL
Response time 20-45 min
Sampling volume 1-500 mL
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Desktop microscope
Company 4-deep
Country Canada
Parameter Particles, bacteria or other organisms
Principle Optical, morfologi
Application Medical (cancer research, urine and blood)
Validation N/A
Measuring unit Particles or cells/mL
Detection limit N/A for drinking water
Installation + flow cell
Response time Immediate
Sampling vol. N/A
Maintenance Every day wiping glass, flushing flow cell
Market Rental, 200,000 DKK + >40,000 DKK for training
Comments Needs operator at least remotely
7000 RMS (RMS-ON90)
Company Mettler Toledo A/S
Country International
Parameter Bacteria and particles (simultaneously)
Principle Optical
Application Pharmaceutical waters
Validation N/A
Measuring unit Bacteria and particles/mL
Detection limit 1 Biocount (no conversion)
Installation at-line
Response time N/A
Sampling volume N/A
Maintenance N/A
Market N/A
Comments US technology just acquired Information not available
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BACMON (Zebra)
Company Grundfos A/S
Country Denmark
Parameter Particles and total bacteria
Principle Optical
Application Drinking water
Validation MS in prep
Measuring unit Cells and particles/mL
Detection limit 160 cells/mL
Installation at-line
Response time 10 min
Sampling volume 6 µL fixed in the flow cell
Maintenance Regular change of flow cell
Market For rent
BACMON
Erfaring: TREFOR VAND A/S, Aarhus Vand A/S, HOFOR A/S
Positivt: stabil drift, god brugerinterface, lille vedligeholdelse
Ulemper: Vanskeligt at tolke resultat,
men fortsat anvendelse
HIOFOR: installeret i ledningsnet, lang opholdstid
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EZ-ATP
Company Applitek
Country Israel, Belgium
Parameter ATP
Principle Chemiluminescence
Application Drinking water and wastewater
Validation N/A
Measuring unit N/A
Detection limit 103 cells/ mL
Installation at-line
Response time 15 min
Sampling volume N/A
Maintenance N/A
Market N/A
Comments No reply
Biocounter
Company Biosensores
Country Spain
Parameter N/A
Principle N/A
Application Wastewater, beverage
Validation N/A
Measuring unit N/A
Detection limit 103 cells/ mL
Installation at-line
Response time 15 min
Sampling volume N/A
Maintenance N/A
Market N/A
Comments No reply
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At-line teknologier under udvikling Name Bactiline N/A MiniLab MicroLab N/A
Company Mycometer A/S SBT Aqua ApS Optiqua (Optisense) Nwater Blusense
Diagnostics ApS
Background Sells Bactiquant test kit for detection of bacteria
in water
Spin-off from DTU Nanotech
Sells the EventLab technology for
chemical contaminant monitoring
N/A Spin-off from DTU
Nanotech
Country Denmark Denmark NL / Singapore Finland Denmark
Parameter Activity of several gram positive and negative
bacteria Total bacteria Total bacteria N/A
Specific bacteria strains
Principle Enzymatic activity and
fluorescence Impedance flow
cytometry
Optical chip inferometry with specific antibody
binding
Optical
Molecular extraction and
detection of specific genes
Validation/ development
stage
Developed prototype(s) are currently validated in the laboratory. To follow
validation at Vitens water-works (NL)
2 months continuous operation at Vilstrup
Vandværk. No measurements, only
robustness of microfluidic system
Manual handling. No test in drinking water. Further development
and automation during 2016/2017
Prototype developed
Prototype for detection of
proteins in blood and bacteria in
urine. Not tested in drinking water
Exp. release 2016 End 2016 2017 2017 End 2016
Comments An automated version of
the Bactiquant test -
Automated combined version of the
EventLab and MiniLab (on market)
- -
Funding EU (FP 7) via
SmartWater4Europe project
MUDP and VTUF EU (H2020) via
aquaSHIELD project N/A VTUF
Information og validering
Mangel på transparens i information fra forhandler
interest is to promote their products.
technical details are not disclosed due to commercialization interests (patent)
arbitrary validation tests
difficult to realise how far these technologies are in the development process
as manufacturing companies tend to be too optimistic about their progress and the expected time of market release.
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Validation
Metoder på forskningsplan
Method Immunoassays Polymerase
chain reaction (PCR)
Fluorescence in situ hybridization
(FISH) Electrical Flow cytometry
Raman spectroscopy
Principle
Antigen-antibody specific binding Used to
capture the target microorganisms and are typically combined with a
detection method
DNA amplification and detection
by dyes or probes
RNA hybridization and microscopic
detection of fluorescent probes
bound in cells
Different methods where
signal is detected by an electrode.
Are typically combined with
immunomagnetic separation
Labelled cells are suspended in a flow stream that passes
through a laser beam89-90
Measurement of inelastically
scattered light after excitation4.
Provides quantitative and
qualitative information
Sub- categories
Fluorescent immunoassays
Immunomagnetic assays Immunosorbent assays
- -
Electrochemical Electrical
impedance Piezoelectric
-
Surface enhanced Raman
spectroscopy (SERS) Lazer
tweezer Raman spectroscopy
(LTRS)
Current application
Various types of water and food to detect E. coli Salmonella Legionella
Enterococci
Various types of water
Groundwater to detect E. coli and
bacteria involved in chromium dioxide
reduction in a contaminated site
E. coli Staphylococcus enterotoxin B Salmonella
Cryptosporidium
Bacteria Yeast Algae Protozoa Viruses
E. coli Streptococcus Enterococcus
Bacteria spores
Advantages
High automation potential Can be
combined with several detection methods and can target a wide range
of microorganisms Potential for lab-on-a-chip implementation
Highly sensitive Specific Rapid Potential to
detect several micro-
organisms in a single reaction Potential for lab-on-a-chip
implementation
Allows the detection of viable but non-culturable
cells Can be combined with flow
cytometry in a microfluidic system
Potential for online and lab-
on-a-chip implementation
Rapid and accurate Various staining and labelling methods An
automated and on-line system and in lab-on-a-
chip systems Implemented in an automated and on-line system and in lab-on-
a-chip systems
Very sensitive Identification of
single bacteria cells Molecular
information with no reagents use Potential for microfluidic
implementation
Dis- advantages
Capturing of non-targeted microorganisms
Rapid but not quantitative Sample
concentration needed
Complex method
Enrichment steps may be required
Piezoelectric sensors require incubation time
Subjective data interpretation Bacteria detection requires high sensitivity of detection instruments High cost
Cells must be immobilised by
physical or chemical contact
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Større igangværende danske og EU projekter
Future water WP 8- Biosensors
Detection of E. coli DNA in distribution
systems
Real-time water quality monitoring by
electrical detection
AQUAWARN SMARTWATER4-EUROPE
Aquavalens Aquashield AQUAVIR
Partners
Ringkøbing-Skjern Forsyning A/S, VIA University College, Amphi-Bac ApS,
Alectia A/S, Minus 10dB ApS
Blusense Diagnostics
SBT AquaVerdo Vand A/S Svendborg Vand A/S
Sønderborg Vandforsyning
A/S Guldborgsund
Forsyning/ Nykøbing
Waterworks
6 EU partners Mycometer A/S and 19 EU partners
DTU FOOD DTU
Environment Højmarklaboratoriet A/S IPU A/S Nordvand A/S and 34 EU
partners
Optiqua (Optisens)
DTU Nanotech Unisensor A/S Delta A/S DHI A/S and 10 EU
partners
Country Denmark Denmark Denmark UK and other EU countries
Several EU countries
Several EU countries
The Netherlands
Several EU countries
Duration Sept 2013 - dec-15 Jul 2015-mar
2016 Jan 2015-sep
2016 Dec 2013-nov
2015 Jan 2014-dec 2017
Feb 2013-jan 2018
Jan 2015-jan 2017
Nov 2013-okt 2016
Relevant aim
Ddevelopment of single
components for alarm, automated
sampling and characterization of a
contamination
Development of a cheap sensor to monitor E.
coli in the distribution
system within 30 min
Further development of the SBT Aqua
sensor
Integrated detection of pollution in water using state-of-the-
art microfluidic technology
Further development
and integration of sensor
technologies
Development of automated
detection methods for
microbial water quality monitoring
Integrated sensor to detection
chemical and microbial
contamination
Development of a
microfluidic automated
virus analyser
Funding VTUF, MUDP VTUF VTUF, MUDP EU (FP7-SME-
2013) EU (FP7-CP) EU (FP7) EU (H2020) EU (FP7-CP)
Funding – EU-projekter
SMARTWATER4EUROPE 10,043,233 EUR
3 år, December 2017.
1 WP: Yderligere udvikling og validering. ‘Bactiline’ fra Mycometer A/S
Aquavalens 11,909,166 EUR
5 år, January 2018.
Udvikling af metoder og teknologier til at påvise pathogener i drikkevand og vand i fødevareproduktion. DTU FOOD og DTU Environment deltager i WP med udvikling af ATP-sensor
aquaSHIELD 1,123,136 EUR
Jan 2017
Udvikling af integreret sensor Optiqua (NL). Kombinere 2 sensorkomponenter: chlorine residual og risikostoffer
AQUAVIR 5,246,429 EUR
3 år, October 2016
Udvikling af bærbar on-site microfluidic system til at påvise virus i ferskvand. Koordineret af DTU Nanotech
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Status i dag? Området i hastig udvikling
Ofte optimistiske forventninger fra udvikler
Ofte begrænset dokumentation
Konkurrencehensyn
Er der tilstrækkelig dokumentation (også objektivt?)
Lovende teknologier, men udfordringer (undervurderet) på markedet:
Høj følsomhed
Kort svartid
Lav vedligeholdelse
Tekniske udfordringer i drikkevand
Få target-organismer
Lav mikrobiel aktivitet
Interferens
Udfældninger
Den ‘ideelle sensor’ er (endnu) ikke helt på plads
Status i dag?
Den ‘ideelle sensor’ er (endnu) ikke helt på plads
Coliforme og E. coli detektion tilgængeligt
Enzymatisk (colilert-baseret)
Responstid: 2-18 timer
Forbrug af reagenser, refill
Alternativer til coliforme og E. coli?
Totalt antal bakterier
~real-tid
Flere detektionsprincipper: optisk, enzymatisk, ATP
Langt i udvikling
Udvikling tager
Tid (5-10 år)
Betydelige investeringer
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Information og validation
lack of transparency in the information provided by the manufacturing companies, as their main interest is to promote their products. Often, detailed validation information or technical details are not disclosed due to commercialization interests. Therefore the information provided by the manufacturing companies should be interpreted with care, especially when conclusions are drawn from arbitrary validation tests and when satisfactory performance in other fields is assumed to apply also for drinking water.
Compiling technical details about technologies under development is even more challenging than the methods currently on the market, because companies are particularly reluctant to disclose any information before they launch their product. Often, it is also difficult to realise how far these technologies are in the development process, as manufacturing companies tend to be too optimistic about their progress and the expected time of market release.
Information og validering
Mangel på transparens i information fra forhandler
interest is to promote their products.
technical details are not disclosed due to commercialization interests (patent)
arbitrary validation tests
difficult to realise how far these technologies are in the development process
as manufacturing companies tend to be too optimistic about their progress and the expected time of market release.
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Funding – EU-projekter
‘SMARTWATER4EUROPE’ 10,043,233 EUR
three-year period ending December 2017107. one specific WP aims to further develop sensor technologies and to validate their use at selected demonstration sites108. The ‘Bactiline’ technology developed by Mycometer A/S
‘Aquavalens’ 11,909,166 EUR
five years and ends January 31st 2018.
develop methods and practices to detect pathogens in drinking water and in water used for food preparation. DTU Environment is involved in a work package that aims to develop an automated platform for detection, based on ATP
‘aquaSHIELD1,123,136 EUR
ends January 1st 2017
further develop an integrated sensor solution developed by the Dutch company Optiqua67. Specifically, the aim of the project is to combine two already developed sensor components for online monitoring, which monitor chlorine residual, and performs rapid screening of a set of high priority threat substances67. The project involves only Optiqua
‘AQUAVIR’ 5,246,429 EUR
three years, and ends October 31st 2016
The project aims to develop a portable, on-site microfluidic system to detect viruses in different freshwater water bodies. coordinated by DTU Nanotech and involves 13 more partners
Større igangværende danske og EU projekter