emerging compounds in wastewater reuse compounds, effect
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Emerging compounds in wastewater reuse – compounds, effect evaluation and removal technologies – an overview
Dr. Norbert Kreuzinger
Technische Universität WienInstitute for Water Quality and Resource Management
Karlsplatz 13/226-1
1040 Vienna – Austria
norbkreu@iwag.tuwien.ac.at2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
Emerging compounds in wastewater reuse – compounds, effect evaluation and removal technologies – an overview
• “emerging compounds” – what are they?
– CECs: Compounds of emerging concern
– Chemicals and other substances that have no regulatory standard and not currently included in routine monitoring programs
– have been recently “discovered” in natural streams (often because of improved analytical chemistry detection levels)
– potentially cause deleterious effects in aquatic life at environmentally relevant concentrations.
– CECs are not necessarily new chemicals. They include pollutants that have often been present in the environment, but whose presence and significance are only now being evaluated.
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
Photo: EPA
CECs - Compounds of emerging concern
• Organic compounds in < µg/l range in waters– Industrial chemicals
solvents; fuels; components; explosives; dyes; plasticisers; …
– Consumer relatedcontrast agents; personal care products; detergents; sweeteners; ...
– Industrial productsrefrigerants; lubricants; surfactants; flame retardants; preservatives; ...
– targeted bioactivitypesticides; biocides; disinfectants; antibiotics, pharmaceuticals; …
• Antibiotic resistance– Bacteria
– Genes 2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
CECs - Compounds of emerging concern
Is the topic of CECs in the water a matter of “fear mongering” in science?
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
CECs - Compounds of emerging concern
IncreasingDiversity ofChemicals
> 140,000
substances on
global market;
30,000 water
relevant
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
Source: CAS
CECs
“Synthetic chemicals as agents of global change”
Bernhardt et al. Front Ecol Environ 2017; 15(2): 84–90
ΔX = (Xt −Xref) ∕ Xref
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
Modified from Bernhardt et al. Front Ecol Environ 2017; 15(2): 84–90
CECs - Compounds of emerging concern
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
CECs - Compounds of emerging concern
• CECs are found in wastewater and receiving waters due to
– More sensitive LODs in chemical analysis – Yes; but
– we have a whole lot of “new” chemicals and substances that were not present 20; 30; 40 years ago
• Question:
– When were the criteria / parameter used today for describing water quality and wastewater treatment defined?
• UWWD: 1991 (-> “young” in terms of reinvestment periods for wwtps)
• WFD: 2000 (later on: review of hazardous substances + watch list)
– Fit for purpose? Still sufficient? Protection targets? Parameters?2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
Effects of CECs – organic trace compounds
• Effects of “traditional” makro - pollutants (C/N/P)
– Oxygen depletion in environmental waters
– Eutrophication
– Acute toxicity (e.g. NH3)
• Effects of “emerging” mikro – pollutants (CECs)
– First indication: hormones • estrogenic activities
• Population stability of fish, molluscs, makro-invertebrates
– No acute effects; rather long-term subtle effects• Not reflected by conventional acute and chronical in vivo tests
• New effect based in vitro tests with various biological endpoints
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
Effects of CECs – organic trace compounds
• Effect based in vivo assays
– Advantages:• Same sample preparation as for chemical analysis (e.g. SPE extraction)
• Suitable for high throughput analysis (robotics)
• Fast and cheap (compared to chemical analysis and in vivo tests)
• No ethical issues
– Selection of possible endpoints:• General toxicity (cytotoxicity always is accessed due to test setup)
• Hormone active (estrogenicity; anti-estrogenbicity; androgenicity, anti-androgenicity, Progesterone receptor; Glucocorticoid receptor …)
• Oxidative stress response; genotoxicity; response to Polycyclic aromatic hydrocarbons; Xenobiotic sensing; Repression of constitutive transcriptional activation
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
Example:
Assessment of 12 WWTPs by in vitro tests in the frameof the Joint Danube Survey
Effects of CECs – organic trace compounds
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
3-10 x EBT >10 x EBT1-3 x EBT< EBT
WWTP ERα anti-AR GR PPARγ PAH Nrf2 PXR
1
2
3
4
5
6
7
8
9
10
11
12
Bioassay Parameter
anti-AR Anti-androgens
GR Glucocorticoid receptor
ERa Estrogens
Nrf2 Oxidative stress
PPARy Peroxisome proliferator
PAH Polycyclic aromatic hydrocarbons
PXR Xenobiotic sensing
EBT Effect based trigger value
Nikiforos Alygizakis, Characterisation of wastewater effluents in the Danube River Basin with target and non-target chemical screening techniques,
in vitro bioassays and antibiotic resistant genes analysis NORMAN – ICPDR Joint Danube Survey 4 (JDS4)
Effects of CECs – organic trace compounds
• Chemical analysis– What substances?
– Looking at parent-substances only. Transformation products?
• Effect based in vivo assays– Broad variety of endpoints available nowadays (not only estrogenicity)
• Combination of both approaches suggested– Whole effluent assessment -> mixture toxicities (substances with same effect)
– Chemical monitoring of tracer substances -> technology working
– Chemical analysis to investigate reason for toxicological effect
In vivo biotests: “New experience” for WWTP regulators and operators
Not suitable for operation control to prove the proper operation of an installed technology but the suitability
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
CECs – antibiotic resistance
• Clinical relevant pathogenic bacteria get resistant to new antibiotics
• Antimicrobial resistance (AMR) is one of the largest health-related issues worldwide, recently acknowledged by the General Assembly of the United Nations in the A/RES/71/3/2016
• Wastewater treatment:
– Close contact between environmental & clinical bacteria
– High density of bacteria
– Resistance genes get exchanged from clinical bacteria to environmental bacteria -> genpool
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
CECs – antibiotic resistance - sewer
Abundance of various antibiotic resistance genes in sewers -Case Study
• Hospital waste water
• Municipal wastewater without hospital
• Similar / same pattern
• Except: 2 ARGs in hospital wastewater only:blaKPC & VanB
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - KreuzingerPM Gabriela Karina Paulus – ANSWER project – KWR – water Cycle Institute – The Netherlands
CECs – antibiotic resistance - wwtp
12 wwtps in Danube catchment
attention: compare results to results from municipal / hospital sewer!
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
Concentration of ARGs [Gene copies/16S rRNA]
PM Nikiforos Alygizakis – ANSWER project – Environmental Institute Slovakia
CECs – removal technology
• Tertiary CAS treatment according to BAT (low loaded AS Systems) does remove target substances to a certain extend; some substances very well –despite not being designed for CEC removal!
• Effluent concentrations for CEC nevertheless > EBT (effect based trigger values)
• Technologies for further elimination are not “new”, but mostly adopted from drinking water treatment
– Membrane technologies - MBR (NF)
– Activated carbon application – AC (PAC, GAC)
– Chemical oxidation – Ozonation – O3 2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
CECs – organic micro pollutants - wwtp
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
• CAS (conventional activated sludge plant)– Same wastewater (=same
location of plant)!
– High loaded C-removal only
– Low loaded Ni/Deni
– Subsequent Ozonation step (zspec 0,7 mg O3/ mg DOC)
Schaar, et al. 2010, Environmental Pollution 158, 1399-1404
CECs – organic micro pollutants - advanced technology
• Options for advanced wwttechnologies “ready to use”
– PAK: powdered AC
– GAK: granular AC filter
– Ozonung: O3 (zspec +/- 0.5)
• Suggested for Austria:Multibarrier with O3 & GAK
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
CECs – organic micro pollutants - ozonation
• Removal of organic trace pollutancs is depending on specific ozone dose!
• zspec = mg O3 / mg DOC
• Competition of organic matrix (mg/l) with trace pollutants (µg/l)
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
CECs – removal of pathogens - ozonation
• removal for pathogens not as dependant on specific ozone dose!
• Similar results for zspec > 0.2 mg/mg
• No removal for C. perfringens spores
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
CECs – organic micro pollutants – GAC Filter
• Breakthrough behaviour Acesulfame K
• Competition of organic matrix (mg/l) with trace pollutants (µg/l)
• Competition adsorption kinetics between substances
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
Bed volumes treated [m3 m
-3]
0 2000 4000 6000 8000 10000 12000
c/c
0 [-]
0.0
0.5
1.0
1.5
2.0
2.5
3.0GAC stand-aloneGAC post-treatment
CECs – response to in vitro biotests - wwtp
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
CALUX-Assays
EBTBEQ LOQ **
FK-AB O3-OUT* N GAC-N FK-AB O3-OUT N GAC-N
Cytotox n.a. 0.51 0.54 0.64 0.53 0.51 0.54 0.64 0.53
ERa 0.3 0.49 0.07 0.05 0.08 0.04 0.06 0.05 0.05
anti-AR 14.4 6.0 5.3 7.2 5.5 5.3 5.3 7.2 5.5
Nrf2 21 107 127 107 64 27 27 27 27
p53 n.a. 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01
p53 +S9 n.a. 220 243 300 240 220 243 300 240
PAH 6.2 170 70 45 55 1.0 1.2 1.0 1.0
PXR 54 53 18 20 65 10.2 6.0 6.2 5.9
If BEQ < LOQ LOQ assumed as result* ± 0.7 g O3/g DOC ** average LOQ of single measurements
CECs – antibiotic resistance - wwtp
FC – CFUsfor 2 C/N/DN/P CAS plants in Austria
• Sul resistance
• Average over 4 samplings
• Inflow (Zulauf) & effluent(Ablauf) 24h samples
• ~ 0,5% resistant FC in inflow
• ~ 0,05% resistant FC in effluent
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
qPCR; results in copies/100ng DNA Culture based
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
CECs – antibiotic resistance – advanced wwtp
Reduction in absolute number of bacteriarelative abundance of genes (ARG & 16s) +/- constant
„in“ = effluent of CAS = inflow of advanced steps
treatment
Summary
• We are living in a changing world; more and more chemicals and substances found in wastewater.
• Todays topics
– Organic trace pollutants
– Antibiotic resistance
• We can neither measure them all in chemical analysis nor access their toxicological potential individually - > whole effluent in vivo assays
• Direct reuse of treated ww & indirect reuse via receiving waters face the same challenges
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
Conclusion
• CAS (conventional activated sludge) aren’t that bad in CEC removal!
• Compared to high loaded C-removal plants, CAS with nitrification have a significant removal of CECs and toxic / biological effects (no significant further removal with Denitrification)
– (They have installed a system immanent online toxicity measurement: nitrification ;-)
• Even advanced treatments do not remove all CECs! (Multibarrier Concept)
• BAT (as low loaded AS plants) are an absolute requirement for implementation of further steps!
– Removal of competing matrix (“DOC”; COD; e.g. humic acids)
– Reduction of O3 depleting substances (e.g. NO2)
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
Conclusion
• Advanced ww treatment by ozonation and GAC-filtration is applicable even on smaller plants with limited personel resources
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
Thank you
For your attention
2018.11.21 - XIV Jornadas Tecnicas de ESAMUR - Kreuzinger
The ANSWER project has received funding from the European
Union’s Horizon 2020 research and innovation programme under
the Marie Skłodowska-Curie grant agreement No 675530.
Dr. Norbert Kreuzinger
Technische Universität WienInstitut für Wassergüte und Ressourcenmanagement
Karlsplatz 13/226-1
1040 Wien
norbkreu@iwag.tuwien.ac.at
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