Combined NEREUS COST Action ES1403 and NORMAN Association meeting

NEREUS COST ACTION ES1403

3-4 December 2015

Rome

Date: 3 December 2015

Participants list:

Name / Surname / Organisation
Despo / Fatta-Kassinos / University of Cyprus, Nireas-IWRC
Célia / Manaia / UniversidadeCatólica Portuguesa
Luigi / Rizzo / University of Salerno
Marie-Noëlle / Pons / CNRS-University of Lorraine
Thomas / Berendonk / TU-Dresden
Christophe / Merlin / CNRS-University of Lorraine
Dimitra / Lambropoulou / Aristotle University of Thessaloniki

This meeting had two major objectives:

  1. Establishment and validation of a protocol for quantification of ARG
  2. Development of an action plan to provide useful feedback to the European Commission for the legislative instrument that is under preparation

Establishment and validation of a protocol for quantification of ARG

Discussion and Suggested Protocol

  1. In order to overcome the limitation with DNA amount available from previous sampling campaigns:
  2. 10 new DNA extracts will be supplied – 5 with low DNA concentration (1-7 ng/ul) and 5 with high DNA concentration (50-100 ng/ul) – these will be supplied by Celia Manaia, Thomas Berendonk, DespoFatta-Kassinos (for low concentration)
  3. Protocols:
  4. Each lab will run all genes, following the protocols recommended by the “original lab” (=ctx, Thomas B; vanA, Thomas Schwartz; qnrS and blaTEM, Celia; sul1, Eddie Cytryn; intIand 16S rRNA gene, Christophe Merlin); and with the reference pNORM (DIGESTED);
  5. If some optimization is needed in each lab: i) all modifications must be informed; ii) the problems and results obtained with the un-optimized protocol must be reported to the colleagues;
  6. Determinations for harmonization will start with the 16S rRNA gene. All DNA extracts will be diluted 1:10, 10:100, 1:1000 and 10 ul of template DNA will be used per reaction. When these analyses are concluded each partner must send it to Celia, who will combine data and will send to all. This stage will provide information on:
  7. qPCR inhibitors
  8. Adjust the bacterial-equivalent DNA (16S rRNA gene copy; dilution and amount of DNA) to use in the ARG determination
  9. Validate 2014 and 2015 data already obtained:
  10. This will be made after all the others and will use the 2014 and 2015 protocols using own references and pNORM (DIGESTED). Each lab will analyse the own genes (=ctx, Thomas B; vanA, Thomas Schwartz; qnrS and blaTEM, Celia Manaia; sul1, Eddie Cytryn; IntIand 16S rRNA gene, Christophe Merlin).
  11. Samples will be the same referred to above (in 1.);
  1. Proposed schedule

January / February / March / April / May / June / July
1
2b,c
2a,b
3
Surface water of 2015

Date: 4 December 2015

Participants list:

Name / Surname / Organisation
Despo / Fatta-Kassinos / University of Cyprus, Nireas-IWRC
Stefan / Kools / KWR Watercycle Research Institute
Lian / Lundy / Middlesex University
Celia / Mania / UniversitàCatolica Portuguesa
Mira / Petrovics / ICRA Catalan Institute for Water Research
Thomas / Letzel / Technische Universität München
Christophe / Merlin / CNRS-University of Lorraine
Marie-Noëlle / Pons / CNRS-University of Lorraine
Tobias / Schulze / Helmholtz Centre for Environmental Research - UFZ
David / Schwesig / IWW Water Centre
Dimitra / Lambropoulou / Aristotle University of Thessaloniki
Benjamin / Lopez / BRGM (Bureau de Recherches Géologiques et Minières)
Jaroslav / Slobodnik / EnvironmentalInstitute, s.r.o.
Luigi / Rizzo / University of Salerno

Development of an action plan to provide useful feedback to the European Commission for the legislative instrument that is under preparation

The objective of this part of the meeting was to discuss and decide on the best waysforthe COST Action NEREUS and the NORMAN Association to contribute to the on-going activities and discussions, on the Water Reuse policy instrument that is under preparation at the European Commission level, and propose exact steps on how the Action can make a significant contribution. A first draft of a Water Reuse guidance document is under preparation as a task of the PoM (Programme of Measures) Working Group within the WFD Common Implementation Strategy (CIS), and it is noted that contaminants of emerging concern including antibiotic resistance are not included. A drafting group has been set up by JRC in which members of the COST Action NEREUS and NORMAN Association participate. Hence, we would like to prepare comprehensive and well-justified suggestions to be considered by the Commission, which is "open" to evaluate proposals for minimum quality criteria for the assessment of the reclaimed water. Use of reclaimed water for agriculture irrigation and groundwater aquifer recharge are currently the key topics.

Most attendees agreed to commit themselves to working towards the development of the policy recommendation for the European Commission (except D. Schwesig and T. Shultze).

D. Fatta-Kassinos underlined the fact that the task force need to be pragmatic and realistic in whatever decided to be developed due to the extremely limited time available (deadline is June 2016). She presented the history of important events/meetings and the background information on the Commission’s activities (presentation attached).

The key points on which the discussion should focus are:

-Decide on whether both wastewater reuse for irrigation and groundwater recharge should be considered.

-A text/table with modes of action/recommended bioassays with human health based trigger values

-A text/table of ‘reuse water’ substances (with or without threshold values) with mechanism for its update (Watch List)

-Text/table on ARBs – relying strongly on input from NEREUS/NORMAN WG5;

-Monitoring schemes (types of reuse water, frequencies etc.).

M. Petrovic mentioned there is a need to be less ambitious so that we can achieve what we set out to do. She proposed to focus solely on groundwater recharge since focusing on both practices would be too ambitious.

J. Slododnik:

  • List of pollutants and the ‘watch list’ should be considered.
  • California’s small list of CECs for groundwater recharge to be considered. Australia has a general statement – PPCPs are a concern and should be looked out for.

Recommendation from T. Schulze to contact: Tamara Grummt (UBA, Germany) relevant to step 1 (modes of action; she is currently writing guidelines).

S. Kools: DEMEAU project: Human health trigger values - acceptable daily intake, mode of action specific (identify target). Express bioassay activity to certain potency – e.g. dioxin concept. Based on CALUX bioassays – can move from response in bioassays to certain allowable threshold (ER-CALUX, AR-CALUX, GR-CALUX, PR-CALUX).

DEMEAU project data presented by J. Slobodnik to the Commission’s WG (also presented in paper – Brand et al., 2014 in Environment International); WG considered it very new and possibly good include CEC in legislation. Also agreed to include AR on the list.

S. Kools: make distinction between WET and specific mode of action.

D. Fatta-Kassinos: how many labs can carry out CALUX? Can WWTP implement these tests? EU will focus on the feasibility of our proposals as well.

L. Rizzo: suggested a two-step control including (i) a first “routine control” (higher yearly frequency depending on the people equivalent size served) with a shorter list of parameters including some CECs, bioassays and easy to measure antibiotic resistance indicators (e.g., E. coliresistant to selected antibiotics) and (ii) a second “check control” with a larger list of parameters (but lower yearly frequency also related to people equivalent size served) eventually including, depending on the results of “routine control” about CECs bioassays and antibiotic resistance E. coli, a highernumber of CECs, bioassays and more specific (genetic) analysis to better characterize antibiotic resistance spread.The suggestion was appreciated by the whole task force.

B. Lopez: can only do groundwater recharge if we can prove that will not negatively impact the groundwater status. How can bioassays be used to prove that the water is safeguarded? Do we want this to be included in the regulations? Interesting to have trigger values – to identify potential issue of concern that needs further investigation. Can we align approaches for groundwater protection and irrigation?

J. Slobodnik: ISO standards with 2 parts - Reuse of wastewater for irrigation and for groundwater recharge. Will use same framework for both – with amendments per purpose. Proposal is to have standards for CEC for groundwater recharge but not for irrigation. Upgrade of Drinking water Directive – will consider a risk-based approach. Also upgrade of UWWT Directive will take place.

M. Petrovic: We need to consider the fact that in case of exceeding the trigger values for effluents for recharging groundwater what is followed? Discharging to rivers? What would be the logic behind this?

B. Lopez: no direct injection of surface water into ground water; occurrence of any anthropogenic substances is a problem.

T. Letzel: 2-step bioassays (as in the US); risk-based origin; easy, fast – not the solution – but a first step.

Summary (D. Fatta-Kassinos and J. Slobodnik):

  • There was a general agreement that CECs should be included in our proposal for legislation. It would be logically inconsistent to address only recharge because irrigation is not only related to crops uptake where there is a gap of knowledge, but it is also a route that can lead to impacts on groundwater, surface water and living organisms.
  • The task force NORMAN+NEREUS ES1403 will deal with both wastewater irrigation of food crops and groundwater recharge.
  • No voluntary measures are supported by the task force.
  • ARB – want to have it in; extend current microbiological parameters with additional data e.g. number of E. coliable to grow on antibiotic supplemented culture medium versus antibiotic-free culture medium (ratio between drug resistant and total Escherichia coli)
  • We would like to include a list of CECs, covering the entire water cycle (link to WFD, DWD, UWWT Directive etc). Keep it short and manageable e.g. 10 substances.
  • We would like to propose further to the above, a watch list. This can help develop a body of evidence e.g. ARB could be a watch list candidate.
  • Need to establish background values of CEC (including ARBs)
  • Monitoring schemes – frequencies, locations of samplings, etc. The team will try to consider a risk-based procedure (as too many scenarios) rather than prescriptive e.g. identify most hazardous control points.