Denitrification barriers as a tool

for nitrogen reduction in agriculture catchments

Bednarek A1/2., Mankiewicz–Boczek J1/2., Gągała I1., Serwecińska L1., S. Szklarek1 , Izydorczyk K. 1, Frątczak W. 1, Zalewski M.1/2

1 European Regional Centre for Ecohydrology of the Polish Academy of Science, 3 Tylna, 90-364 Lodz, Poland; E-mail: agnik@biol.uni.lodz.pl2 Department of Applied Ecology, Faculty of Biology and Environmental Protection, University of Lodz, 12/16 Banacha, 90-264 Lodz;

Groundwater nitrates concentrations in European Union countries, 2008-2011

4-3 Extracts from the report on implementation of the EU Nitrates Directive, 2008-2011

Poland

limit - 50 mg of nitrates per liter in groundwater (Nitrates Directive, European Commission, 1991)

emission NH3

Fot. http://en.wikipedia.org Fot. M.Ubraniak

Fot. M.Wysocki

NITROGEN POLLUTIONDIFFUSED POINT

leaching NO3

mineral fertilizers

lack of ecotone zone

Approximately 50% of N comming from fertilisers

is not taken up by plants

and is going to deeper soil layers and to the groundwater

Natural denitrification process:

• In grassland soil denitrification decrease below - 0.6 m

• In arable land denitrification decrease below - 0.4 m

(Zwart, 2004)

DENITRIFICATION

2NO3- à 2NO2

- à 2NO à N2O à N2(N 5+ ) (N 3+ ) (N 2+ ) (N 1+ ) (N 0 )

• one of the most important process in bioremediation

• depends on availability of organic carbon!

The aim of the study was to test denitrifying ditches, composed with different organic carbon sources – harl flax, pinus sawdust, straw and a mix of brown coal and calcium coal.

DENITRIFICATION

2NO3- 2NO2

- 2NO N2O N2

optimum pH 7,0 - 8,2

The final products depends on pH:

> 7.3 N2

< 7.3 NO i N2O

“Denitrification bed” – containers filled with a material rich in C (Schipper et al., 2010b). They can be put in stream to clean water in small rivers or drainage system (Robertson and Merkley,

2009) or on the outflow from point source of nitrate (Schipper et al., 2010a; Warneke et al., 2011a; Warneke et al., 2011b).

(Schipper et al., 2010b)

Methods based on denitrification process, ussually applied in the field can be divided into three groups:

“Denitrification wall” - material rich in C arranged vertically in shallow ground water perpendicular to the flow of these waters (Schipper et al., 2010b).

Denitrification walls are useful to prevent area (Schipper and Vojvodic-Vukovic, 2000) and point (Bednarek et al., 2010) pollution of nitrate.

“Denitrification layer” – are horizontal layer of material rich in C (Schipper et al., 2010b).

Problem in Poland - deposition of manure directly on the ground (especially in medium-sized farms of 10-15 ha)

Concentration of nitrate (NO3-)

in groundwater between 300 mg/l to 2000 mg/l

Długość ≈ 15 mObjętość dodanych trocin ≈ 10 m3(30%)

disposal of manure

manure storage site after constructing the ditch

– denitrifying barrier

Solution…

(Bednarek et al., 2014)

The obligation to have manure plates are covered by the farm:

• carrying out the raising of poultry above 40 000 places or breeding pigs over 2 000 places weighing more than 30 kg (or 750 places for cows ) (the Act on fertilizers and fertilization, 2007)

• on designated under the Nitrates Directive prone areas the outflow of nitrogen from agricultural sources ( Regulation of the Minister of Environment of 23 December 2002 )

• in other cases dung plate must build only farmers who have been granted some EU grants ( Sectoral Operational Programme and the Rural Development Programme )

Different sources of organic carbon used in denitrifying barrier

lignite -brown coal pine sawdust barley straw harl linseed

ditch denitrifying barrierPBS1/A8/2012 MIKRAZO

POLAND

P1 Non point sources – zb. Czarnocin

Point souces–Uniejów

Point sources –Jerwonice - A

Non point sources – Tresta

N R14 0061 06/ 2009 GEOWŁÓKNA - Development of model geofibrous, biodegradable, biological deposits for recultivation of nitrogen and phosphorus in threatened areas of agricultural landscape

Point sources –Jerwonice - B

Construction of the denirification barrier

228,3

34,6

10,8

I II III0

50

100

150

200

250

300

NO 3

[mg

. l-1]

Average ± Standard deviation

Volume of added pinus sawdust≈ 10 m3(30%)

Phot.1. Restoration of a point source of nitrogen (Demo site, Jerwonice village) (Bednarek et.al. 2014a)

before ditches in ditches behind ditches-400

-200

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

mg N

O3 l

-1

Average Average +/- SD Min-Max

Fig. 1. Average (and +/- DS and Min/Max value) nitrate concentration in ground water before, in and behind ditches in Jervonice demonstration site (from: Bednarek et. al. 2014a).

Czarnocin Reservoir

NO3 concentration in water of drainage canal - 90,8 mg/l

NO3 concentration in ground water - 46,8 mg/l

Concentration of nitrate (NO3-)

in groundwater between < 200 mg/l

Czarnocin Reservoir - constructionof the denitrification barrier

Harl flax

befor

after

No landscape deformation

Source of pollution Organiccarbon

Average nitrogenreduction (%)

The maximumnitrogen load

(mg/l)

Themaximumnitrogen

reduction (%)

Diffused

Point

farmland Harl flax 50 90 NO3- 88,0

farmland Mix sawdustand straw 22 97,9 NO3

- 58,9

cow manure Brown coal 65 > 2000 NO3- 84,8

cow manure Harl flax 51 339 NO3- 94,8

pig manure sawdus 85 361 NO3- 95

Results….

Results….

• The higher the concentration of incomming pollution the better the work bed , and NO3 reduction rate ( also in winter !)

The Pilica River:Overall length: 342 kmMean discharge: 18,51 m3/s

The Pilica River Catchment:Total area: 9 245 km2

Area covered by LIFE+ EKOROB Project: 4 933 km2

Land use: agricultural - 64 %forest - 31%

THE PILICA RIVER CATCHMENTGlobal Reference Site for UNESCO IHP Ecohydrology

and LTER-Europe LTSER Platform

(Corine Land Cover 2006)

(www.geoportal.gov.pl)

B1

B4B5

0

100

200

300

400

B1 B4 B5

NO

3 co

ncen

trat

ion

ingr

ound

wat

er[m

g/l]

Polluted water as per Nitrates Directive

Denitrification wall

Planting or/ and harvesting plants

Recreation facilities (jetty)

Nitrogen pollutions

(Izydorczyk, Frątczak, Zalewski 2011)

Highly effective buffer zonesfor reduction of diffuse nitrogen pollutions

Średnia Min-Maks

upstream the wallwithin the wall

downstream the wall0

20

40

60

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100

120

Denitrification wall in buffer zone

- pilot results

NO

3co

ncen

trat

ion

in g

roun

dwat

er [m

g/l]

Polluted water as per Nitrates

Directive

Next Question…

What is the optimum carbon substrate for activation of microorganisms?

Main objective: increase the efficiency and acceleration of the activation of denitrifying barriers

Study supported by the National Centre for Research and Development, No. PBS1/A8/2012MIKRAZO - Microbial activators in denitrifying deposits used for the treatment of nitrate pollutionfor the implementation of the Water Framework Directive and the Nitrates Directive.

1 2

Environmental samples from denitrifying barrier (red color)

Soil samples - unculturable denitrifying bacteria

Isolation from the environment& cultivation

Culturable denitrifying bacteria

Material and methods - bacteria characterization, isolation and application

storage of manure

as a source of nitrogen

compounds

Genetic analyses of soil samples

PBS1/A8/2012 MIKRAZO

Scheme of bioreactors settings and options to fill them by substrate, and inoculation an additional pool of microorganisms

1 CDB2

1

without carbon substrate

with mix straw/lignite

1 - without additional pool of bacteria2 - with culturable denitrifying bacteria (CDB)3 - mix of environmental bacteria

MIX 3

PBS1/A8/2012 MIKRAZO

Result … experiment with mix straw/lignite (C substrate) and different variants of bacteria

1 - without additional pool of bacteria; 2 - with culturable denitrifying bacteria (CDB); 3 - mix of environmental bacteria

0200400600800

control 1 2 331st Jul 2014 (after 6 days of experiment)Conc

entr

atio

n [m

g/l]

NO3- NO2- NH4+

0200400600800

control 1 2 37th Aug 2014 (after 12 days of experiment)

Conc

entr

atio

n [m

g/l]

NO3- NO2- NH4+

0200400600800

Control 1 2 310th Sep 2014 (after 45 days of experiment)

Conc

entr

atio

n [m

g/l]

NO3- NO2- NH4+

sewage

SewageNO3 - - 448 mg/l; NO2 - - 730 mg/l and NH4+ - 260 mg/l

sewage

sewage

without carbon substrate mix straw/lignite

mix straw/lignite

mix straw/lignite

without carbon substrate

without carbon substrate

After 6 days

After 45 days

After 12 days

reduction of NH4+

reduction of NO3- reduction of NO3

- reduction of NO3-

Artificial sewage:

ØNH4 – 362 mg/LØNO3 – 822 mg/L

Study supported by the National Centre for Research and Development, No. PBS1/A8/2012MIKRAZO - Microbial activators in denitrifying deposits used for the treatment of nitrate pollutionfor the implementation of the Water Framework Directive and the Nitrates Directive.

0

100

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500

600

średnia z eksperymentów

stęż

enie

azo

tanó

w [m

g N

O3-

/ L]

kontrola

złoże bez bakterii

złoże z bakteriami hodowalnymi

złoże z bakteriami terenowymi

In sewage : 873 mg NO3-/L NO3 concentration

[mg / L]% of reductioncontrola 383without additional pool of bacteria; 111 71%with culturable denitrifying bacteria (CDB); 116 70%mix of environmental bacteria 144 62%

Redukcja azotanów w poszczególnych złożach denitryfikacyjnych(experyment Tresta)

Vilage Mikołajów k. ŁodziUsed carbon source : straw + lignite

MIKRAZO PBS1/A8/2012

Field experiment….

PBS1/A8/2012 MIKRAZO

Nitrogen removal in denitrification barriers in Mikołajów winter time

MIKRAZO PBS1/A8/2012

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above the barrier in the barrier below the barrier

nitrate mg/l nitrite mg/l ammonium mg/l

Denitrification - the organic C is the electron donor and NO3- acts as the electron acceptor; full process - reduction of NO3- by microorganisms to dinitrogen gas (N2)

MIKRAZO PBS1/A8/2012

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nitrate mg/l nitrite mg/l

Nitrogen removal in denitrification barriers in Mikołajówspring time

PBS1/A8/2012 MIKRAZO

Mikrobiologiczne aktywatory w złożach denitryfikacyjnych stosowanych do oczyszczania zanieczyszczeń azotanowych dla wdrażania Ramowej Dyrektywy Wodnej i Dyrektywy Azotanowej

PBS1/A8/5/2012

Odciek…. NO3

mg/lNO2

mg/lNH4 mg/l

23.11.2015 30,2 8,2 2401,9

30.11.2015 139,4 11 2432,8

11.12.2015 355,4 2,5 889,3

Mikrobiologiczne aktywatory w złożach denitryfikacyjnych stosowanych do oczyszczania zanieczyszczeń azotanowych dla wdrażania Ramowej Dyrektywy Wodnej i Dyrektywy Azotanowej

PBS1/A8/5/2012

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A1 A2 B1 B2 C2 A2 B2 C2 A1 A2 B1 B2 C2 A1 A2 B1 B2 C2 A1 A2 B1 B2 C2 A1 A2 B1 B2 C2

Azotany [mg/L]

Azotyny [mg/L]

Amonowe [mg/L]

• The application of methods based on enhancement of denitrificationprocess create an opportunity to increase the efficiency of ground water protection in

the ecosystem scale.

• In the catchment of intensive farming or pasture, around the point source, e.g. storage manure, or near a coastline, denitrification ditches seem to be the most appropriate solution.

• The highest denitrification effect was observed at ditches constructed around the point-sources egz. storage manure, due spring and autumn period (even above 95% of nitrogen reduction).

• In the case of denitrification ditches construction there is no landscape deformation.Construction the manure plates permanently change the landscape.

• These biotechnology seems to be the alternative solutions for building the manureplates. It is important to use easily-obtained, locally-available carbon-rich materialsto minimize transport costs.

Main conclusions

Thank you for invitation and hospitality

•Environmental Protection AgencyRepublic of Serbia Ministry of Agriculture and Environmental Protection

• Institute For Nature Conservation of Serbia

•Chamber of Commerce and Industry of Serbia

Study supported by :

N R14 0061 06/ 2009 GEOWŁÓKNA - Development of model geofibrous, biodegradable, biological deposits for recultivation of nitrogen and phosphorus in threatened areas of agricultural landscape

PBS1/A8/2012 MIKRAZO - Microbial activators in denitrifying deposits used for the treatment of nitrate pollution for the implementation of the Water Framework Directive and the Nitrates Directive

LIFE08 ENV /PL/000519EKOROB: Ecotones for the reduction of diffuse pollution

Thank you for your attention !

PBS1/A8/2012 MIKRAZO

Dr Kinga KrauzeLong-Term Ecosystem

Research, ecosystem services

Prof. Joanna Mankiewicz- BoczekEcohydrological biotechnologies & ecotoxicology

Dr hab. Katarzyna Izydorczyk, EH biotech for nonsource pollution

& reservoirs eutrophication

Dr Magdalena UrbaniakEmerging pollutants

Dr Ilona Gągała,EH Molecular methods

Prof. Małgorzata GodlewskaIchtiology & echsounding

Prof. Maciej ZalewskiEcohydrology Theory

Prof. Piotr FrankiewiczFish biol/Biomanipulation

Dr Tomasz Jurczak, Urban River rehabilitation &

toxicology

Dr Liliana Serwecińska,Microbiology/Biotech

Dr Iwona Wagner,Urban Ecohydrology

Dr Agnieszka Bednarek, Agricultural biotechnology &

denitrification

Dr Edyta KiedrzyńskaRiver floodplain and point

sources pollution