PROJECT FINAL REPORT
Grant Agreement number: 299382
Project acronym: DON_HYPO
Project title: The role of dissolved organic nitrogen (DON) on the development and extend of
eutrophication-driven hypoxia and responses to global warming
Funding Scheme: FP7-MC-IEF
Period covered:
from
1st May 2012
to 30th April 2014
Name of the scientific representative of the project's co-ordinator1, Title and Organisation: Prof.
Daniel Conley LUNDS UNIVERSITET
Tel: +46462220449
Fax: +46462224830
E-mail: Daniel.Conley@geol.lu.se
Project websiteError! Bookmark not defined. address:
1
Usually the contact person of the coordinator as specified in Art. 8.1. of the Grant Agreement.
4.1 Final publishable summary report
Executive Summary
Coastal hypoxia is a mounting problem worldwide that has been recognised as a major threat to
marine coastal biota. The decline in dissolved oxygen can affect ecosystems structure and
functioning. Managerial efforts to prevent hypoxia and recover ecosystems that have already been
affected by hypoxia are largely based on nutrient reduction plans. Most nutrient reduction plans
are focused on the reduction of the inorganic fraction of the nutrient loadings, particularly in
reductions of nitrate and phosphate inputs. However, the concentration of dissolved organic
nitrogen (DON) frequently exceeds that of dissolved inorganic nitrogen (DIN) in both marine and
freshwaters. Recent studies indicate that many components of the DON pool can play an active
role in supplying, directly or indirectly, nitrogen nutrition to phytoplankton and bacteria and may
affect community metabolic rates and species composition of the ambient microbial assemblage.
Elucidate the role of DON in the development and extend of eutrophication-driven hypoxia is
essential to set successful management strategies in order to reduce hypoxia development and
recover ecosystems experiencing hypoxia at present, to better protect marine biodiversity.
Summary Description of Project
The main objective of the DON-Hypo project was to evaluate the role of DON in planktonic and
bacterial community metabolic rates in the Baltic Sea (the largest coastal area suffering from
eutrophication-driven hypoxia) and to evaluate the effects of global warming on the interaction
between DON inputs, metabolic rates and oxygen dynamics.
We have contributed to the main goal of the project by:
1) Determining the DON content of a selected coastal station in the Baltic Sea 2) Determining the
DON inputs from different sources to estimate the availability and reactivity of DON from various
sources (municipal wastewater treatment plant effluents, atmospheric deposition, rivers) 3)
Assessing the metabolic rates responses from both whole planktonic and bacterial coastal marine
communities to DON additions from different sources 4) Assessing metabolic rates responses
from both whole planktonic and bacterial coastal marine communities to the combined effect of
DON inputs and warming 5) Assessing the chemical composition of DON from different sources
and from different seasons on coastal waters, as well as biological utilization of specific
compounds after 7-days incubations
We have sampled monthly 3 selected rivers during one year (Lyckebyån, Emån and Umeälven) at
different latitudes of Sweden and representative of different land-use patterns. The sampling effort
has been higher than previously proposed by the project, as it has been coordinated with a wider
sampling of the selected rivers. Atmospheric deposition and water from a municipal wastewater
treatment plant have been sampled 4 times each since the beginning of the project. Results derived
from this sampling will be submitted in the near future to the open journal Biogeosciences.
Eight experiments to assess metabolic rates responses to DON additions have been performed in
August 2012, January 2013, April 2013, May-June 2013, July 2013, August 2013, November
2013 and December 2013. Results from one of these experiments have been presented at the
ASLO 2013 Aquatic Sciences Meeting. Additionally, results from 2 experiments have been
presented in a seminar at the Aquatic Ecology department (Lund University).
Two experiments to assess metabolic rates responses to combined effect of warming and DON
additions have been performed in summer (September 2013) and winter (March 2014). Results
from one of these experiments have been presented as a poster in the Ocean Sciences meeting
2014, held on Honolulu on February 2014.
We have implemented diverse water analysis, such as Dissolved Primary Amines (DPA) analysis,
Total Dissolved Nitrogen (TDN), ammonium (NH4), Bacterial production, Chlorophyll a content
and inorganic nutrients analysis with an automated analyser. All samples analyses from
experiments and sources have been performed: a total of 738 samples have been analysed for
DPA, TDN, inorganic nutrients and ammonium; 666 for chlorophyll a content and 250 for BP.
We have also implemented a Matlab program to calculate metabolic rates from oxygen data
measured continuously with Pre-Sens oxygen sensors.
Description of the main S&T results
The main results obtained with the present project are:
-Monthly data over a year on dissolved organic nitrogen, inorganic nutrients, TDN, DPA,
ammonium, chlorophyll a concentration, and flow from 3 different Swedish rivers representative
from different land-use patterns and different latitudes. These results will be submitted to the
open-access peer-review scientific journal Biogeosciences in a short future.
Figure 1. Dissolved primary amines (DPA) content on the three different rivers along one year.
Figure 2. Total dissolved Nitrogen (TDN) concentration for the three different rivers along one year.
Figure 3. Nitrate (NO3-) concentration for the rivers Lyckebyån, Emån and Ume along one year.
Figure 4. Nitrite (NO2-) concentration for the 3 rivers along one year.
Figure 5. Ammonium (NH4+) concentration for the 3 rivers along one year.
Figure 6. Phosphate (PO43-) concentration for the 3 rivers along one year.
-Metabolic rates of whole planktonic and bacterial communities for a total of 8 experiments with
5 different treatments performed on duplicates. Data for TDN, DPA, Gross Primary Production
(GPP), Net Community Production (NCP) and Community Respiration (CR), DON, Bacterial
Production (BP), inorganic nutrients, pigment composition, ammonium, dissolved organic carbon
(DOC) and chlorophyll a have been analysed in duplicate for all experimental bottles.
-CR and BP, as well as TDN, DPA, DON, inorganic nutrients, DOC, ammonium for a 2 warming
experiments conducted on the dark.
River
No addition
Waste water
4
Ln CR
2
0
-2
-4
39.5
40.0
40.5
41.0
41.5
42.0
1:kT
Figure 7. Results of mixed-effects models showing the effect of temperature (1/kT) on planktonic
respiration. To account for temporal pseudo-replication, the statistical model includes sampling dates as
fixed factors. Black dots represent data from seawater, white dots samples with DON addition from Rivers,
and red dots samples with DON addition from Waste Water treatment Plants (WWTP).
60
% Increase CR
50
40
30
20
10
0
Seawater
River
WWTP
Figure 8. Changes in Community Respiration rates (CR) calculated from Q10 values derived in the
warming experiments with a 4ºC water warming.
-Activation energies and Q10 values for community respiration and gross primary production
derived from the relationship between metabolic rates and temperature for all the experiments.
8
No addition
River
Waste water
Inorganic nutrients addition
6
Ln CR
4
2
0
-2
-4
40.0
40.5
41.0
41.5
42.0
1:kT
Figure 9. Results of mixed-effects models showing the effect of temperature (1/kT) on planktonic
respiration. To account for temporal pseudo-replication, the statistical model includes sampling dates as
fixed factors. Black dots represent data from seawater, white dots samples with DON addition from
Rivers, red dots samples with DON addition from Waste Water treatment Plants (WWTP) and blue dots
samples with inorganic nutrients additions.
8
No addition
River
Waste water
Inorganic nutrients addition
6
Ln GPP
4
2
0
-2
-4
41.0
40.5
40.0
41.5
42.0
1:kT
Figure 10. Results of mixed-effects models showing the effect of temperature (1/kT) on gross primary
production (GPP). To account for temporal pseudo-replication, the statistical model includes sampling
dates as fixed factors. Black dots represent data from seawater, white dots samples with DON addition
from Rivers, red dots samples with DON addition from Waste Water treatment Plants (WWTP) and
blue dots samples with inorganic nutrients additions
30
% increase CR
% increase GPP
25
% increase
20
15
10
5
0
Seawater
IN
River
WWTP
Figure 11.Changes on metabolic rates (Gross Primary Production (GPP) and Community Respiration
(CR)) with a 4ºC water warming calculated from Q10 values derived from experiments conducted at in
situ temperature.
-A total of 55 samples have been analysed to determine the specific chemical composition of
sources of DON to coastal waters, as well as in marine coastal water and at the beginning and end
of the experiments, to assess bioavailability of DON.
The main conclusions obtained from the above-mentioned results are:
The results obtained with the present project support the prediction that warming of the Baltic Sea
will increase planktonic respiration rates faster than planktonic primary production. Higher
increases in respiration rates than in production may lead to depletion of the oxygen pool, further
aggravating hypoxia in the Baltic Sea.
DON inputs have been found to have a synergistic effect with temperature, further increasing
respiration rates with warming.
Nutrient reduction plans should also include the organic fraction of the nutrients, specifically
DON and have into account the synergistic effects with warming.
Potential impact
Predictions on effects of global warming and eutrophication in dissolved oxygen dynamics in near
and far future are needed to elucidate the consequences of the lack of oxygen on biodiversity and
ecosystems structure and functioning. The results of this project could help better protect marine
ecosystems, preserve biodiversity and set successful management targets for coastal waters.
Results obtained here will be published in peer-review scientific journals and a press release will
be made to inform citizens and policy makers on the importance of taking into account the organic
fraction of the nutrients when designing nutrient reduction plans aimed to reduce eutrophicationdriven hypoxia on coastal areas, specially in the Baltic Sea.
4.2 Use and dissemination of foreground
Results from the present project will result in four publications that will be submitted in the near
future. The manuscripts under process are:
Vaquer-Sunyer, R., Reader, H. E., Pinhassi, J., Conley, D. and Kritzberg, E. Effects of DON
inputs from rivers and wastewater treatment plants to metabolic rates in the Baltic Sea. To be
submitted to Estuaries and Coasts.
Vaquer-Sunyer, R., Reader, H. E., Conley, D. and Kritzberg, E. River DON inputs to the Baltic
Sea. To be submitted to Biogeosciences.
Vaquer-Sunyer, R., Pinhassi, J., Conley, D. and Kritzberg, E. Planktonic metabolic rates
responses to warming and additions of Dissolved Organic Nitrogen (DON). To be submitted to
Biogeosciences.
Reader, H. E., Vaquer-Sunyer, R., Kritzberg, E, Conley, D. and Koch, B. Bioavailability and
chemical characterization of dissolved organic nitrogen (DON) form Swedish rivers and waste
water treatment plants effluents determined by Fourier transform ion cyclotron resonance mass
spectrometry.
Results have been presented at two international conferences as well as in a seminar at the
Aquatic Ecology department from Lund University.
Vaquer-Sunyer, R. and Conley D. J. Effects of dissolved organic Nitrogen (DON) inputs on
planktonic metabolism in the Baltic Sea. ASLO meeting 2013. New Orleans, USA. 18-22
February 2013. Oral communication.
Vaquer-Sunyer, R., Conley D.J. and Kritzberg, E. Effects of warming and Dissolved Organic
Nitrogen (DON) inputs on planktonic metabolism in the Baltic Sea. 2014 Ocean Sciences
Meeting, Honolulu, Hawaii, USA. 23-28 February 2014. Poster.
Vaquer-Sunyer, R. and Conley, D.J. The role of dissolved organic Nitrogen (DON) on the
development and extend of eutrophication-driven hypoxia and responses to global warming.
Aquatic Ecology seminar. Lund University, Lund, Sweden. 6 March 2013.
Dissemination activities related to the project:
-Interview in a local journal “Es Busqueret” (Mallorca, Spain) on winter 2013. Attached here.
- Article in popular press: “Coastal dead-zones on the rise” on “The conversation”. Published the
25th June 2013. http://theconversation.com/coastal-dead-zones-on-the-rise-15496
TEMPLATE A1: LIST OF SCIENTIFIC (PEER REVIEWED) PUBLICATIONS, STARTING WITH THE MOST IMPORTANT ONES
NO.
Title
1
2
3
4
2
Main
author
Title of the
Number, date or
periodical or
frequency
the series
Publisher
Place of
publication
Year of
publication
Relevant
pages
Permanent
identifiers2
(if available)
Is/Will
open
access3
provided to
this
publication
?
yes
Planktonic
metabolic rates
responses to
warming and
additions of
Dissolved
Organic
Nitrogen (DON)
River DON
inputs to the
Baltic Sea
VaquerSunyer,
Raquel
To be
submitte
d to
Biogeosc
iences
In
progres
s
VaquerSunyer,
Raquel
In
progres
s
yes
Effects of DON
inputs from
rivers and
wastewater
treatment plants
to metabolic
rates in the
Baltic Sea
Bioavailability
and chemical
characterization
of dissolved
VaquerSunyer,
Raquel
To be
submitte
d to
Biogeosc
iences
To be
submitte
d to
Estuaries
and
Coasts
In
progres
s
No
Read
er,
Heath
er E.
In
progres
s
A permanent identifier should be a persistent link to the published version full text if open access or abstract if article is pay per view) or to the final manuscript accepted for publication (link to
article in repository).
3 Open Access is defined as free of charge access for anyone via Internet. Please answer "yes" if the open access to the publication is already established and also if the embargo period for open
access is not yet over but you intend to establish open access afterwards.
organic nitrogen
(DON) form
Swedish rivers
and waste water
treatment plants
effluents
determined by
Fourier
transform ion
cyclotron
resonance mass
spectrometry
TEMPLATE A2: LIST OF DISSEMINATION ACTIVITIES
Countries
addressed
NO.
Type of activities4
Main leader
1
Conference talk:
“Effects of
dissolved organic
Nitrogen (DON)
inputs on
planktonic
metabolism in the
Baltic Sea”
Seminar: “The
VaquerSunyer, R.
2
Vaquer-
Title
Date/Period
ASLO
18-22 February
meeting 2013. 2013.
Aquatic
6 March 2013
Place
New
Orleans,
USA
Lund
Type of
audience5
Scientific
Community
Scientific
Size of
audience
International
Sweden
4
A drop down list allows choosing the dissemination activity: publications, conferences, workshops, web, press releases, flyers, articles published in the popular press, videos, media
briefings, presentations, exhibitions, thesis, interviews, films, TV clips, posters, Other.
5 A drop down list allows choosing the type of public: Scientific Community (higher education, Research), Industry, Civil Society, Policy makers, Medias, Other ('multiple choices' is
possible).
3
4
5
role of
dissolved
organic
Nitrogen
(DON) on the
development and
extend of
eutrophicationdriven hypoxia
and responses
to global
warming”
Conference
poster:
“Effects of
warming and
dissolved
organic
nitrogen
(DON) on
planktonic
metabolism in
the Baltic Sea”
Interview
Article in
popular press:
“Coastal deadzones on the
rise”
Sunyer,
R.
Ecology
seminar
University,
Lund,
Sweden
Community
VaquerSunyer, R.
Ocean
Sciences
Meeting
2014
23-28 February
2014.
Honolulu,
Hawaii, USA
Scientific
Community
International
VaquerSunyer, R.
VaquerSunyer, R.
Es Busqueret
Winter 2013
Mallorca,
Spain
Civil Society
Spain
The
conversation
25 June 2013
Civil Society
International
Section B (Confidential6 or public: confidential information to be marked clearly)
Part B1
TEMPLATE B1: LIST OF APPLICATIONS FOR PATENTS, TRADEMARKS, REGISTERED DESIGNS, ETC.
Confidential
Click on
YES/NO
Type of IP
Rights7:
Foreseen
embargo
date
dd/mm/yyyy
Application
reference(s)
(e.g.
EP123456)
Subject or title of
application
None
6
Note to be confused with the "EU CONFIDENTIAL" classification for some security research projects.
7
A drop down list allows choosing the type of IP rights: Patents, Trademarks, Registered designs, Utility models, Others.
Applicant (s) (as on the application)
4.3 Report on societal implications
Replies to the following questions will assist the Commission to obtain statistics and
indicators on societal and socio-economic issues addressed by projects. The questions are
arranged in a number of key themes. As well as producing certain statistics, the replies will
also help identify those projects that have shown a real engagement with wider societal
issues, and thereby identify interesting approaches to these issues and best practices. The
replies for individual projects will not be made public.
A General Information (completed automatically when Grant Agreement number is
entered.
Grant Agreement Number:
Title of Project:
Name and Title of Coordinator:
B Ethics
299382
The role of dissolved organic nitrogen (DON) on the
development and extend of
Prof.
Daniel Conley hypoxia and responses to
eutrophication-driven
global warming
1. Did your project undergo an Ethics Review (and/or Screening)?
 If Yes: have you described the progress of compliance with the relevant Ethics Review/Screening
Requirements in the frame of the periodic/final project reports?
No
Special Reminder: the progress of compliance with the Ethics Review/Screening Requirements should be
described in the Period/Final Project Reports under the Section 3.2.2 'Work Progress and Achievements'
2.
Please indicate whether your project involved any of the following issues (tick
box) :
RESEARCH ON HUMANS
 Did the project involve children?
 Did the project involve patients?
 Did the project involve persons not able to give consent?
 Did the project involve adult healthy volunteers?
 Did the project involve Human genetic material?
 Did the project involve Human biological samples?
 Did the project involve Human data collection?
RESEARCH ON HUMAN EMBRYO/FOETUS
 Did the project involve Human Embryos?
 Did the project involve Human Foetal Tissue / Cells?
 Did the project involve Human Embryonic Stem Cells (hESCs)?
 Did the project on human Embryonic Stem Cells involve cells in culture?
 Did the project on human Embryonic Stem Cells involve the derivation of cells from Embryos?
PRIVACY
 Did the project involve processing of genetic information or personal data (eg. health, sexual
lifestyle, ethnicity, political opinion, religious or philosophical conviction)?
 Did the project involve tracking the location or observation of people?
RESEARCH ON ANIMALS
 Did the project involve research on animals?
 Were those animals transgenic small laboratory animals?
 Were those animals transgenic farm animals?
YES
 Were those animals cloned farm animals?
 Were those animals non-human primates?
RESEARCH INVOLVING DEVELOPING COUNTRIES
 Did the project involve the use of local resources (genetic, animal, plant etc)?
 Was the project of benefit to local community (capacity building, access to healthcare, education
etc)?
DUAL USE

Research having direct military use
 Research having the potential for terrorist abuse
No
No
C Workforce Statistics
3.
Workforce statistics for the project: Please indicate in the table below the number of
people who worked on the project (on a headcount basis).
Type of Position
Number of Women
Experienced researchers (i.e. PhD holders)
Scientific Coordinator
PhD Students
Other
1
Number of
Men
1
4. How many additional researchers (in companies and universities) were recruited
specifically for this project?
Of which, indicate the number of men:
0
D Gender Aspects
5.
X

Did you carry out specific Gender Equality Actions under the project?
Y
e
s
N
o
6. Which of the following actions did you carry out and how effective were they?
Not at all
effective




X
Very
effec
tive
Design and implement an equal opportunity policy

Set targets to achieve a gender balance in the workforce

Organise conferences and workshops on gender

Actions to improve work-life balance

Researcher and co-ordinator participated in a gender based organization based at
Other:
Lund University
7. Was there a gender dimension associated with the research content – i.e. wherever people were the
focus of the research as, for example, consumers, users, patients or in trials, was the issue of gender considered and
addressed?
Yes- please specify

X
No
E Synergies with Science Education
8.
Did your project involve working with students and/or school pupils (e.g. open days,
participation in science festivals and events, prizes/competitions or joint projects)?
Yes- please specify

X
No
9. Did the project generate any science education material (e.g. kits, websites, explanatory
booklets, DVDs)?
Yes- please specify

X
No
F Interdisciplinarity
10.
Which disciplines (see list below) are involved in your project?
Main discipline8: 1.5


Associated discipline8: 1.4
Associated discipline8: 1.3

G Engaging with Civil society and policy makers
11a
Did your project engage with societal actors beyond the research
community? (if 'No', go to Question 14)

X
Yes
No
11b If yes, did you engage with citizens (citizens' panels / juries) or organised civil society (NGOs,
patients' groups etc.)?
8
Insert number from list below (Frascati Manual).




No
Yes- in determining what research should be performed
Yes - in implementing the research
Yes, in communicating /disseminating / using the results of the project

11c In doing so, did your project involve actors whose role is mainly to organise
X
the dialogue with citizens and organised civil society (e.g. professional mediator;
communication company, science museums)?
12.
Did you engage with government / public bodies or policy makers (including
international organisations)
X



No
Yes- in framing the research agenda
Yes - in implementing the research agenda
Yes, in communicating /disseminating / using the results of the project
13a Will the project generate outputs (expertise or scientific advice) which could be used by
policy makers?
Yes – as a primary objective (please indicate areas below- multiple answers possible)

Yes – as a secondary objective (please indicate areas below - multiple answer possible)
X
No

13b If Yes, in which fields?
Agriculture
Audiovisual and Media
Budget
Competition
Consumers
Culture
Customs
Development Economic and
Monetary Affairs
Education, Training, Youth
Employment and Social Affairs
Energy
Enlargement
Enterprise
Environment
External Relations
External Trade
Fisheries and Maritime Affairs
Food Safety
Foreign and Security Policy
Fraud
Humanitarian aid
Human rights
Information Society
Institutional affairs
Internal Market
Justice, freedom and security
Public Health
Regional Policy
Research and Innovation
Space
Taxation
Transport
Yes
No
13c If Yes, at which level?
Local / regional levels

National level
X
European level
X
International level

H Use and dissemination
14. How many Articles were published/accepted for publication in
peer-reviewed journals?
None
To how many of these is open access9 provided?
None
How many of these are published in open access journals?
None
How many of these are published in open repositories?
None
To how many of these is open access not provided?
Please check all applicable reasons for not providing open access:
 publisher's licensing agreement would not permit publishing in a repository
 no suitable repository available
 no suitable open access journal available
 no funds available to publish in an open access journal
 lack of time and resources
 lack of information on open access
 other10: ……………
15. How many new patent applications (‘priority filings’) have been made?
0
("Technologically unique": multiple applications for the same invention in different jurisdictions
should be counted as just one application of grant).
16. Indicate how many of the following Intellectual
Property Rights were applied for (give number in each
box).
Trademark
0
Registered design
0
Other
0
17. How many spin-off companies were created / are planned as a direct
result of the project?
none
Indicate the approximate number of additional jobs in these companies:
18. Please indicate whether your project has a potential impact on employment, in comparison
with the situation before your project:

In small & medium-sized enterprises
 Increase in employment, or

Safeguard
employment,
or
In large companies


None of the above / not relevant to the project
 Decrease in employment,
 Difficult to estimate / not possible to quantify
Indicate figure:
19. For your project partnership please estimate the employment effect
resulting directly from your participation in Full Time Equivalent (FTE = one
2
person working fulltime for a year) jobs:
9
Open Access is defined as free of charge access for anyone via Internet.
10
For instance: classification for security project.

Difficult to estimate / not possible to quantify
I Media and Communication to the general public
20. As part of the project, were any of the beneficiaries professionals in communication or media
relations?
Yes

X No

21. As part of the project, have any beneficiaries received professional media / communication
training / advice to improve communication with the general public?
Yes
 No
X
22 Which of the following have been used to communicate information about your project to the
general public, or have resulted from your project?

Coverage in specialist press
 Press Release
Media
briefing
X
Coverage in general (non-specialist) press



X


TV coverage / report
Radio coverage / report
Brochures /posters / flyers




DVD /Film /Multimedia

Coverage in national press
Coverage in international press
Website for the general public / internet
Event targeting general public (festival, conference,
exhibition, science café)
23 In which languages are the information products for the general public produced?

X

Language of the coordinator
Other language(s)

English