© 2024 The Authors Water Reuse Vol 14 No 1, 1 doi: 10.2166/wrd.2024.

081

Potential and risks of water reuse in Brandenburg (Germany) – an interdisciplinary case

study

Daniel Dittmann a,*, †, Alina H. Seelig b,†, Mogens Thalmann c,†, Theresa Wilkesa,d, Veikko Junghans e,

Daniel Zahn b, Sondra Klitzke a, Andre Peters c,f, Jens Haberkamp d, Thorsten Reemtsma b

and Aki S. Ruhl a,g

a
German Environment Agency, Schichauweg 58, Berlin 12307, Germany
b
Helmholtz Centre for Environmental Research – UFZ, Permoserstr. 15, Leipzig 04318, Germany
c
Technische Universität Braunschweig, Universitätsplatz 2, Braunschweig 38106, Germany
d
Münster University of Applied Sciences, IWARU, Corrensstr. 25, Münster 48149, Germany
e
Humboldt-Universität zu Berlin, Unter den Linden 6, Berlin 10099, Germany
f
Thünen Institute of Agricultural Technology, Braunschweig 38116, Germany
g
Technische Universität Berlin, Straße des 17. Juni 135, Berlin 10623, Germany
*Corresponding author. E-mail: [email protected]
†
These authors contributed equally in writing this manuscript.

DD, 0000-0003-2793-7787; AHS, 0009-0005-6000-987X; MT, 0009-0004-1587-1244; VJ, 0000-0003-3356-3558; DZ, 0000-0003-3038-9371;
SK, 0000-0001-5895-0074; AP, 0000-0002-8893-8102; TR, 0000-0003-1606-0764; ASR, 0000-0002-2443-4722

ABSTRACT

For Brandenburg, a region in Germany with increasing water shortage and drought events, water reuse can counteract competition scenarios
between drinking water supply, agricultural irrigation, and industrial use. Centralized and decentralized sources for reclaimed water are found
to potentially substitute 245 or 28% of irrigation water, respectively, in agriculture production in Brandenburg. For such a reuse scenario, the
fate of organic micro-pollutants is examined for diatrizoate (DZA) and carbamazepine (CBZ). Retention in local sandy soil and transfer into
roots and leaves of arugula are analyzed in lysimeter studies and greenhouse pot experiments. Vertical transport was found for DZA and
accumulation in or on arugula roots with a root concentration factor of 1,925 + 34% but a low bioconcentration factor due to intrinsic mol-
ecule properties. CBZ was not found to be mobile in the sandy soil but accumulates in arugula roots and leaves by factors of 70 + 7% and
155 + 12%, respectively. Further research on potential plant uptake and groundwater enrichment for more substances is highly rec-
ommended as well as tertiary wastewater treatment prior to water reuse.

Key words: diatrizoic acid, domestic wastewater, emerging contaminants, farmland, organic trace substance

HIGHLIGHTS

• The volume of treated wastewater exceeds irrigation water by a factor of 2.5 in Brandenburg.
• Water reuse could reduce competition between drinking water supply, industry and agriculture.
• Organic micro-pollutant pathways for sandy soil to groundwater and to plants are assessed.
• Carbamazepine did not reach 10 cm depth in lysimeter studies probably due to acidic soil pH.
• Diatrizoate obtained very high concentrations in arugula roots but were low in leaves.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC BY 4.0), which permits copying, adaptation and
redistribution, provided the original work is properly cited (http://creativecommons.org/licenses/by/4.0/).

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 Water Reuse Vol 14 No 1, 2

GRAPHICAL ABSTRACT

INTRODUCTION
Exploiting wastewater as irrigation water to counteract water stress is a widely applied option for water reuse in arid regions
of the world (Shoushtarian & Negahban-Azar 2020). This is already being implemented extensively, for example in Israel
(Reznik et al. 2017) or California (Cooley & Phurisamban 2016). The use of reclaimed water (RW) counteracts competition
among agriculture irrigation, drinking water supply, and industrial use. Hence, stressed fresh water resources are thought to
be protected ( Jaramillo & Restrepo 2017; Lavrnić et al. 2017; Schwaller et al. 2021), and benefits in terms of ecological costs
can be accounted for (Passarini et al. 2014).
Water reuse comes along with potential hazards to human health through pathogen contamination of the irrigation water
(Amorós et al. 2010; Amahmid et al. 2023). However, decades of experience and the development of quantitative risk assess-
ments demonstrated water reuse to be a secure practice (ISO 2020; Wencki et al. 2020). If multibarrier principles are applied,
which are composed of natural (e.g. reservoirs, soil passage), technical (e.g. filtration, disinfection), and administrative sys-
tems (e.g. risk assessment in the catchment, monitoring of connections to the sewage network), microbiological risks can
be minimized effectively (Mohr et al. 2020).
Anthropogenic trace substances such as pharmaceuticals, pesticides, or industrial chemicals, posing long-term chemical risks,
are less regulated (Helmecke et al. 2020; Shoushtarian & Negahban-Azar 2020). Most of these organic micro-pollutants (OMPs)
are only insufficiently eliminated by wastewater treatment plants (WWTPs) with secondary treatment using activated sludge for
reductions of chemical oxygen demand, nitrogen, and phosphorous (Margot et al. 2015; Alygizakis et al. 2020). Hence, persistent
OMPs remain in secondary treated wastewater or irrigation water, respectively, and can accumulate in soil and plant material or
contaminate groundwater, posing risks to ecosystems and human health. Dependent on OMPs’ intrinsic molecule properties,
composition of the soil, cultivated plants, and environmental and climatic conditions of a region, the fate of individual OMPs
in environmental compartments is determined (Wu et al. 2015). The pathways are mainly divided in transfer of OMPs from
soil to groundwater and from soil to plant, thus potentially entering drinking water sources and the food chain.
Ben Mordechay et al. (2021) analyzed the concentrations in RW for irrigation, soil, and commercially grown crops for 65
OMPs at 400 fields in Israel and reported that primarily OMPs’ concentrations in RW determine the levels detected in soil

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 Water Reuse Vol 14 No 1, 3

and agricultural products. They found that green leaves of RW-irrigated plants obtain higher OMP concentrations than har-
vest organs of root crops (e.g. carrot and potato) or harvested fruits (e.g. tomato and banana), as also reported in previous
studies (Wu et al. 2015; Christou et al. 2019).
The antiepileptic carbamazepine (CBZ) is a wastewater indicator substance ( Jekel et al. 2015) and is well studied for plant
uptake (Shenker et al. 2011; Marsoni et al. 2014; Chuang et al. 2019). The bioconcentration factor (BCF) and the root con-
centration factor (RCF) are defined as ratios between OMP concentrations (ng g
1) in plants or roots, respectively, and in soil
(McKone & Maddalena 2007) and are reported for CBZ between one and several hundreds. CBZ is, therefore, recommended
as a reference for every study on this subject (Wu et al. 2015). Furthermore, CBZ can be transformed by soil micro-organisms
and plants (Christou et al. 2019). The transformation product (TP) 10,11-epoxycarbamazepine accumulates in leaves and is a
potentially genotoxic compound with an acceptable daily intake factor of 1,000 lower than that of the parent compound CBZ
(Malchi et al. 2014; Paz et al. 2016). Non-ionic substances have been reported to transfer into plants more easily (Malchi et al.
2014), while ionic substances are probably retarded in the phloem (Goldstein et al. 2014).
Human biomonitoring studies in Israel (Schapira et al. 2020; Ben Mordechay et al. 2022) revealed elevated CBZ concen-
trations in urine of men, women, and children, most of them being vegetarians consuming vegetables irrigated with RW.
Schapira et al. (2020) pointed out the need to resolve the contradiction between recommended daily vegetable consumption
and the chemical risks of RW irrigation.
Risk assessments often determine the thresholds of toxicological concern (TTC) with differing results depending on OMP
concentrations and irrigated crops. On the one hand, negligible health risks are reported for CBZ (BCF . 100) in maize, rice,
ryegrass, and wheat (Delli Compagni et al. 2020) as well as for diclofenac (BCF . 100 after 3 years) in tomatoes through
uptake from RW after tertiary treatment and disinfection (Christou et al. 2017). On the other hand, comparable low CBZ
concentrations (0.5 μg L
1) lead to elevated 10,11-epoxycarbamazepine concentrations in the leaves of carrots and sweet
potatoes. The TTC value of 62.5 ng kg
1 for children of 25 kg was found to be exceeded by eating 25 g of carrot leaves or
90 g of sweet potato leaves per day, which is common in parts of Asia and Africa. This does not imply direct toxic effects
but indicates the need for detailed toxicity analysis on this OMP. Moreover, there are other OMPs, e.g. lamotrigine with a
TTC value of 2.5 ng kg
1 for children that is exceeded by eating half a carrot (60 g) per day (Malchi et al. 2014).
Previous studies conclude that more research is highly recommended, regarding different agricultural products, substances,
soils, and climatic conditions to properly manage related risks (Helmecke Fries & Schulte 2020). The European regulation on
minimum requirements for water reuse is applied since June 2023 (EU 2020) and is in line with worldwide regulations and
guidelines on water reuse providing no restriction for OMPs in spite of recommending their consideration in the site-specific
risk management (Shoushtarian & Negahban-Azar 2020).
The current study, therefore, focuses on a sandy soil typical for the state of Brandenburg. This is one of the driest regions in
Germany, with an annual mean rainfall of 580 mm and extreme droughts in recent summers. The annual climatic water bal-
ance (Figure 1) is negative with
52 mm by a mean temperature of 9.7 °C (1.4 °C in winter, 18.4 °C in summer). In order to
investigate whether water reuse is a viable option to mitigate competition for water in our case study region, we studied
(a) potential sources for RW in Brandenburg and irrigation demand,
(b) the transfer pathway soil to groundwater for the iodinated X-ray contrast agent diatrizoate (DZA) that is comparably large
in size and CBZ (as indicator OMP) in disturbed soil lysimeters, and
(c) the transfer pathway soil to plant also for DZA and CBZ in greenhouse pot experiments with arugula.

The obtained interdisciplinary perspectives are interconnected through the same sandy soil and RW used for irrigation,
which allows for common evaluation of separately analyzed pathways soil to groundwater and soil to plant. Since the
focus is first on potential water sources and second on the risks of OMP transfers, agricultural yields are not assessed and
no nutrient deficiency experiments are conducted.

MATERIALS AND METHODS

Analysis of the potential of water reuse for Brandenburg
Wastewater generated in Brandenburg is classified in centralized (sewage network and WWTP) and decentralized disposal,
subdivided in cesspit tanks and septic tanks (small-scale WWTP). Respective amounts are officially reported for 2019 (MLUK
2021) and a mean water consumption of 106 L d
1 and person (Destatis 2018) was used for calculations. The actual use of

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Figure 1 | Annual climatic water balance of Germany (left) and the state of Brandenburg (right) over the period from 1991 to 2021. Colors
indicate the local water balances, calculated from precipitation minus potential evapotranspiration. Based on data from the German Climate
Data Center (CDC 2023).

irrigation water was reported lastly for 2009 (Berlin-Brandenburg 2012) and, therefore, needed to be projected based on the
agriculturally used area in 2019 (Destatis 2021). Calculations were done for the growing season (April–October) and for the
whole reference year 2019.
Furthermore, fertilizer application was analyzed for potential substitution by remaining nitrogen (N) and phosphorous (P)
in RW. Due to lack of available data, the mean value of domestic sales volume in 2019–2021 was used to estimate applied
fertilizer amounts (Destatis 2018). Human excreta are calculated with 11 g N and 1.8 g P contents per day and person.

Reclaimed water and soil characteristics

RW was taken from the effluent of a large WWTP in Berlin with secondary treatment and UV-disinfection between May 1 and
September 30. Each batch was analyzed for OMP concentrations immediately after collection from the WWTP, and the
stored RW was reanalyzed to evaluate potential OMP losses. Accompanying chemical parameters for RW are summarized
in Table 1.
The sandy soil was taken from an agricultural field in Brandenburg (north east Germany, 52°160 34.3″N 13°040 39.7″E),
which is located in a region that was formed in the last ice age as a terminal moraine. The soil material was taken from
four different layers (Table 2), with layer A representing the top soil of horizon A and all subsequent layers representing
the mineral horizon.

Table 1 | Characteristic parameters for RW used for irrigation

Base parameters Anions Cations

pH EC NO

3 PO3

4 Cl SO2

4 Kþ Naþ þ
Ca2þ þ
Mg2þ

1
1
1
μS cm mg L mg L

n 5 5 11 15 3 6 15 15 15 15
Mean 7.5 1,212 37.8 0.59 155.4 110.6 84.9 28.5 11.3 112.7
SD 0.2 66 2.9 0.21 7.7 7.9 16.0 1.9 1.2 7.3
Notes: n is the number of measurements of RW batches, EC is the electrical conductivity representing the salinity of RW, and SD is the according standard deviation.

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 Water Reuse Vol 14 No 1, 5

Table 2 | Physico-chemical properties of the four layers of sandy soil

Textural fractions (%)a

Depth Bulk density Corg


Layer cm Texture class Sand Silt Clay g cm ³ % pHH2O pHCaCl2

Ab 0–30 loamy Sand 83.3 11.5 5.3 1.53 0.73 5.4 4.9
L1 31–46 loamy Sand 87.4 7.9 4.7 1.72 0.16 5.7 5.4
L2 47–80 loamy Sand 89.6 7.0 3.4 1.70 0.15 5.8 5.4
L3 81–100 sandy Loam 71.4 17.2 11.5 1.75 0.07 6.4 5.9

1
Notes: Corg is the organic carbon content in wt.%. pH values measured in ultrapure water and 0.01 mol L CaCl2 with a soil solution ratio of 1:2.5.
a
Analyzed by the PARIO method (Durner & Iden 2021).
b
Determined for pot experiments: 0.57% Corg; 0.65% Ct; 0.06% Nt, and 4.98 cmolc kg
1 CECpot.

Lysimeter studies
Two lysimeters (1 m2 cross-sectional area and 1 m depth) were filled with sandy soil according to Table 2 and planted with
summer barley during the vegetation periods 2021 (May 26–August 25) and 2022 (March 7–July 14) and lay fallow in the
interim. Mineral fertilizer was applied in two applications during each vegetation period to optimally supply plants with nutri-
ents. Accordingly, 8, 2.6, and 10 g m
² of N, P, and K was applied in 2021, and 8.1, 1.3, and 5 g m
² N, P, and K in 2022,
respectively. Therefore, no effect of fertilization by the RW was considered in the experiments. All cumulated fluxes at the
upper boundary during the total experimental period (May 26 2021–November 1 2022) of the lysimeters are displayed in
Figure 2. The total inflow of 1,594 L m
2 is comprised of 850 L m
2 precipitation, 543 L m
2 RW irrigation, and
200 L m
2 tap water irrigation. Precipitation was measured by a near tipping bucket corrected after Richter (1995). The
potential evapotranspiration accounted for 1,500 L m
2. During the vegetation period, RW and tap water irrigation were
demand-based. In addition, groundwater recharge was simulated by irrigation with RW along the winters 2021/22 and
2022/23. Hence, seepage water could be collected all over the year. Each lysimeter is equipped with nine glass suction
cups (ecoTech Umwelt-Messsysteme GmbH, Bonn, Germany), inserted every 10 cm. One to four times a month, the seepage
water was sampled by applying a suction of 400 hPa for at least 4 h on all suction cups. Furthermore, samples were collected
from a suction array at 100 cm depth (constant suction of 70 hPa), and of the free draining water at the lower boundary pas-
sing the suction array, which were sampled regularly depending on the quantity accrued (twice a week to once a month).
Mass balances for OMPs are estimated by the volume and concentration of the influent (irrigation) and the effluent (suction
array and free drainage water).

Greenhouse pot experiments

Arugula (Eruca sativa var. ‘Speedy’) was cultivated in the greenhouse (artificial light for 12 h d
1, mean temperature 18 + 2 °C,
mean relative humidity 49 + 13%) by using standard ‘Mitscherlich pots’ (max. vol. 6,200 mL, diameter 20 cm, height 21 cm,
see Figure 3, right), five plants per pot, each filled with 6,500 g sieved (2 mm) and homogenized air-dry agricultural topsoil
(see Table 2, Layer A). All pots were regularly watered to a maximum field capacity of 84%, using the RW, with three repli-
cates. Arugula was watered by simulating drip irrigation, where leafy parts of the arugula did not have any contact with the
irrigation water (in comparison to overhead or sprinkler irrigation). To avoid unwanted growth depression, an initial fertili-
zation was given to all pots (Table 3).
Analyzed arugula was harvested after 65 days at BBCH1 stage 59. Harvested plant material was dried at 30 °C and hom-
ogenized afterward with a ceramic ball mill for further analyses. Soil samples were taken from each vegetated pot
immediately after harvest and were air dried.

Extraction of samples of reclaimed water, soil, and plants

The water samples were stored at
20 °C until sample preparation, then centrifuged for 10 min at 5,000 rpm at room temp-
erature and for an additional 10 min at 13,000 rpm at 4 °C, and subsequently analyzed by direct injection.

1
Biologische Bundesanstalt, Bundessortenamt und CHemische Industrie.

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 Water Reuse Vol 14 No 1, 6

Figure 2 | Cumulated upper boundary fluxes of lysimeters during the experimental period. The outflowing flux potential evapotranspiration
(ET pot) is given in absolute values for better comparison with the inflowing fluxes.

Figure 3 | Lysimeters with summer barley (left) and pot experiments with arugula in the greenhouse (right).

The solid–liquid extraction for soil and plant samples was adapted from a method by Riemenschneider et al. (2017). The
roots and leaves of the arugula plant were processed individually by weighing 0.5 g of soil or plant material in a 15 mL cen-
trifuge glass. Extraction was performed twice using a mixture of methanol and Milli-Q water (1:1, v:v). Following the protocol
of Riemenschneider et al. (2017), an aliquot of the two combined supernatants of both extractions were centrifuged for
10 min at 13,000 rpm and 4 °C.

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 Water Reuse Vol 14 No 1, 7

Table 3 | Initial nutrient application of the pot experiments

1
Nutrient mg kg dry soil Applied as

N 400 Ca(NO3)2 · 4 H2O

P 119 KH2PO4
K 150
Mg 60 MgSO4 · 7 H20

Analysis and quantification

In lysimeter studies, OMPs were analyzed by high-performance liquid chromatography (HPLC) coupled to a triple-quadru-
pole mass spectrometer (MS/MS) as described in more detail by Zeeshan et al. (2023). The analyzed OMPs were
quantified using an internal calibration with isotopic labeled standards.
OMP analyses for the pot experiments were performed by using supercritical fluid chromatography (SFC) (Waters Acquity
UPC2 system) coupled to a triple-quadrupole mass spectrometer (Waters TQXS) using a Ethylene Bridged Hybrid (BEH)
column based on a method by Schulze et al. (2020). For quantification of CBZ and DZA in RW samples, external calibration
was used. The apparent recoveries were included by spiking the samples with the reference standards. For soil and crop
samples, matrix-matched calibrations using non-treated soil and arugula material were used. In addition, for soil samples,
apparent recoveries were included analogous to the water samples.

RESULTS AND DISCUSSION

Potential of water reuse for Brandenburg
About 2.5 million people live in the state of Brandenburg (Germany) on an area of 29,650 km². About 46% of the area is used
for agriculture while about 37% is covered by forest. As shown in Figure 1, large regions have a negative climatic water bal-
ance, additionally suffering droughts in summer. Water irrigated demand for agriculture in Brandenburg was calculated to be
23.1  106 m3 in 2019 (average 72.2 mm) while about 56.6  106 m3 total wastewater is generated in the growing season from
April to October (Figure 4). This wastewater could potentially be reclaimed and could substitute irrigation water withdrawn
from surface and groundwater.
About 11% of Brandenburg’s inhabitants are not connected to sewage networks. A total of 75,000 use septic tanks for
wastewater disposal and 205,000 use cesspit tanks for wastewater storage. Vacuum trucks are regularly collecting the
stored wastewater and transport it several kilometers to the next sewage network connection. These decentralized sources
for RW alone could theoretically supply one-fourth (28%) of irrigation water needs in 2019 (Figure 4). This may offer potential
for innovative point-of-use reuse concepts fitting to local socio-economic structures (UBA 2021).
Furthermore, RW add loads of nutrients to the field and can therefore substitute parts of fertilization. We estimate that
about 68,500 t N (as ammonia) and 3,400 t P (as phosphorus pentoxide) are applied to the fields in Brandenburg each
year. With regard to the untreated wastewater in cesspit tanks, amounts of 4,300 t N and 1,000 t P accrue during a growing
season, which could substitute theoretically a maximum of 6% N fertilizer and 28% P fertilizer. With respect to much lower
concentrations in the effluent of WWTPs (exemplarily calculated with data from Table 1), only 145 t N and 150 t P with sub-
stitution potential of 0.2% N fertilizer and 4.4% P fertilizer, respectively, would have been reached by replacement of the
irrigation water with RW in 2019.
The calculated (maximum) potentials for irrigation water substitution appear to be promising. The concomitant fertilizer
substitution potentials by dissolved nutrients in RW are a positive side effect and worth considering in fertilization planning.
At the same time, negative effects on plants could arise from salts contained in RW (Ofori et al. 2021). An electrical conduc-
tivity (EC) of 1,212 μS cm
1 found in the analyzed RW (cf. Table 1) indicates a slight to moderate restriction for salt-sensitive
plants in arid or semi-arid regions (Ayers & Westcot 1985). However, this does not apply for our case study region and, there-
fore, EC is not restricted. The perspective of irrigation water demand and available RW sources indicates water reuse as a
viable option for Brandenburg to counteract water scarcity and competition. Putting this into practice would require further
consideration of infrastructure, transportation, and water treatment, especially for the decentralized sources. However, risks
by dissolved contaminants (OMPs) need to be assessed with another perspective.

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Figure 4 | Water balance during the growing season (April–October). Accounted for substitution potential for irrigation water used in 2019
with decentralized sewage sources and total wastewater in Brandenburg, Germany.

Pathway soil to groundwater – lysimeter studies

To study potential transfer of OMPs into groundwater, two lysimeters were exemplarily evaluated for DZA and CBZ over a
16-month period (July 2021–October 2022). The temporal concentration courses in different depths were similar for both lysi-
meters (Figure 5).
DZA and CBZ concentrations in the used RW batches for irrigation ranged between 0.6 and 3.3 μg L
1 and 0.3 and 1.4 μg L
1,
respectively (Figure 5, top ‘RW’). After the vegetation period of 2021, the seepage rates increased and DZA was successively
transported down the soil profile, clearly visible by the temporally shifted concentration peaks along the lysimeter depths
(Figure 5, z ¼ 10–90 cm). The concentration decreases with increasing soil depth due to diffusion and hydrodynamic dis-
persion. Five months after the start of RW irrigation, DZA reaches the lower boundary (i.e. the effluent) of the lysimeters
(Figure 5, bottom). The applied DZA mass, from July to August 2021, has left the lysimeters at the lower boundary until
April 2022. These observations suggest that neither sorption nor degradation play an important role for the fate of DZA in
sandy soils. These findings are supported by our observations in laboratory batch experiments, where neither degradation
nor sorption was observed (data not shown). Moreover, a similar behavior of DZA was found in other studies. Ternes
et al. (2007), for example, observed a very persistent and mobile behavior for DZA in lysimeters also filled with a sandy
soil and suggested that DZA could be used as conservative wastewater tracer in soils and groundwater. Kalsch (1999) and
Haiß & Kümmerer (2006) investigated biodegradability of DZA in activated sludge and also found that DZA is persistent.
Interestingly, the DZA concentration peaks in the upper layers (up to 40 cm) during the vegetation period in 2022 are
roughly two times higher than that in RW concentrations. This phenomenon is explained by high evapotranspiration in
the vegetation period (Figure 5, green area in the top subplot ‘RW’), when water leaves the system due to vaporization of
root water uptake and DZA remains in the soil. In particular, the increased concentration in 40 cm depth suggests that
DZA is not or only partly taken up by the barley plants.
Generally, CBZ concentrations were very low in all depths (Figure 5, z ¼ 10–90 cm). Solely in 10 cm depth, values above
the limit of quantification (LOQ) of 5 ng L
1 were detected during the irrigation periods, due to macropore transport. These
findings suggest that CBZ is either strongly sorbed to the mineral or organic surfaces and/or microbially transformed within

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Figure 5 | Top row: concentrations of DZA and CBZ in the RW batches (first subplot) used for irrigation; rows 2–5: concentrations in the
suction cup samples of four selected depths. Each vertically dotted line in the ‘RW’ subplot equals an irrigation event of 10 L. The green area
marks the vegetation period of summer barley. Bottom: total cumulated flux in (influent) and out (effluent) of the lysimeters.

the first few centimeters of the soil. Hence, none of the CBZ applied with the irrigation water reached the effluent. This
assumption is consistent with conducted laboratory batch experiments under aerobic conditions, in which especially sorption
reduced the concentration of CBZ in the soil solution (data not shown). Several other studies reported minor biodegradability
for CBZ under aerobic condition classifying it as a persistent substance in the vadose soil zone (Li et al. 2013; Grossberger

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et al. 2014; Thelusmond et al. 2018); however, König et al. (2016) reported anaerobic transformation of CBZ during bank
filtration. Ternes et al. (2007) observed that sorption and/or biodegradation took place to a certain extent during the passage
of CBZ through lysimeters. However, in their experiments, degradation/retardation of CBZ during the soil passage was rela-
tively low and CBZ was detected in all sampled depths of their lysimeters. Similar to the findings of Ternes et al. (2007), Paz
et al. (2016) also detected CBZ in all depths of their lysimeters (1 m depth), one filled with a sandy loam and another with
loamy sand soil. CBZ sorption is known to be mainly governed by soil organic matter (Paz et al. 2016), whose quality might
change at different pH values (e.g. due to protonation and deprotonation of functional groups). Thus, the contradictive find-
ings may be explained by soil pH, since Ternes et al. (2007) and Paz et al. (2016) reported neutral pH values and our
examinations show slightly acidic soil pH (Table 2). Accordingly, the environmental fate of CBZ can differ strongly due to
site-specific effects related to soil properties.
To this state, no threshold concentrations or maximum loadings exist for OMPs regarding the soil to groundwater transfer
pathways in German legislation2. However, the regulatory aspects of RW for irrigation are frequently discussed, and environ-
mental and health quality criteria with regard to OMP ought to be developed (Helmecke et al. 2020).

Pathway soil to plant – greenhouse pot experiments

To study plant uptake after irrigation with RW, pot experiments were performed in a greenhouse using arugula as a model
plant. DZA and CBZ concentrations were quantified in RW, soil, arugula roots, and leaves (Figure 6). Both DZA and
CBZ could be detected in every compartment above their LOQ. Concentrations in RW were fluctuating slightly around
1.5 (DZA) and 1 μg L
1 (CBZ) and are comparable to the data shown in the lysimeter studies above (Figure 5), since the
same source of RW was used.
DZA was found in highest concentrations in arugula roots between 2,400 and 6,700 ng g
1 dry weight (d.w.), whereas
much lower concentrations of around 70 ng g
1 d.w. were quantified in arugula leaves. This is also shown by the calculated
root and BCFs, demonstrating that DZA is taken up by the roots but not transported to the leaves (Table 4). However, as
discussed elsewhere (Castan et al. 2023), anionic chemicals like DZA are only partially absorbed due to the negatively
charged cell membranes of the roots and the resulting repulsive forces. This implies that DZA may not pass through the
root epidermis and is only attached to the root surface. In contrast, concentrations in the soil material were significantly
lower around 2 ng g
1 d.w. This demonstrates the poor sorption potential to soil surfaces as well as the mobile character
of DZA transporting the compound with the water toward the plant. These data are in line with the results from the lysimeter
studies above, showing that DZA tends to be strongly transferred vertically with the water and can potentially reach ground-
water (cf. Figure 5).
Another OMP analyzed was CBZ, which is well studied and can thus act as an indicator substance. CBZ was identified in
low concentrations in the soil material (,1 ng g
1 d.w.). As reported elsewhere (Scheytt et al. 2006; Martínez-Hernández

Figure 6 | Concentrations of CBZ and DZA in RW used for irrigation and in soil and plants, separated in arugula roots and leaves, referred to
d.w., after harvest.

2
Federal Soil Protection and Contaminated Sites Ordinance of Germany from 1999.

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Table 4 | Mean values of BCFs and RCFs of DZA and CBZ referred to the soil material irrigated with RW

DZA CBZ

BCF [–] 18 + 4 155 + 19

RCF [–] 1,925 + 664 70 + 5

et al. 2016), sorption of CBZ is low. These results stand in contrast to those shown in lysimeter studies above since a removal
of CBZ in the very upper soil layers was shown. This can result from a removal of CBZ by biodegradation in soil and plant
material, e.g. TPs of CBZ were reported in several crops (Malchi et al. 2014; Martínez-Hernández et al. 2016; Riemenschneider
et al. 2016). In addition, it is shown that through the uptake of RW, increased concentrations of CBZ were found in both plant
materials in similar levels at around 71 (arugula leaves) and 32 ng g
1 d.w. (arugula roots). Goldstein et al. (2014) reported
comparable results for plant uptake of CBZ irrigated with treated wastewater containing similar concentrations of CBZ. Also,
the RCF (70) and BCF (155) for CBZ show an uptake by arugula.
In comparison, these two chemicals showed different uptake capacities by the plant. While for DZA a BCF of 18 (less
uptake) was calculated, CBZ has a much higher potential for plant uptake (BCF ¼ 155). This can possibly result from its
ionic state, since non-charged molecules, like CBZ (pH 7.5), may more readily cross cell membranes.
In addition, the maximum amounts of fresh arugula that would still be tolerated from a health perspective for the deter-
mined concentrations of DZA and CBZ were calculated. Since both DZA and CBZ are classified as Cramer class III
(Malchi et al. 2014; Neumann & Schliebner 2019), a TTC value of 1.5 μg/kg bodyweight and per day was used (Kroes
et al. 2004). For DZA (cf.w. ¼ 6.68 ng g
1), an adult with 70 kg bodyweight could eat 16 kg of arugula per day, while for
CBZ (cf.w. ¼ 11.80 ng g
1), 9 kg arugula per day could be eaten to remain below the threshold. These values are comparable
to the ones from Malchi et al. (2014). The amounts stand in no relation to the daily amount of fresh arugula a person normally
consumes and thus are negligible for an adult from a single substance perspective.

CONCLUSIONS
In this study, we showed interdisciplinary examinations on water reuse providing perspectives that are necessary to elucidate
potentials and risks. Our case study region, Brandenburg (Germany), possesses centralized and decentralized sources for RW.
Effluent from centralized WWTPs could substitute 245% while decentralized cesspit and septic tanks could substitute 28% of
water used for agricultural irrigation in 2019. Furthermore, fertilizers could partly be substituted by nutrients in RW from
cesspit tanks, and storage options during winter time may increase the potential. Water reuse is, therefore, a viable option
to mitigate competition for water in Brandenburg. However, implementation requires further consideration of infrastructure,
transportation, and water treatment, especially for decentralized sources. As a limitation, it must also be considered that mini-
mum discharge from WWTPs may be a requirement for streams.
The perspectives of OMP transfer pathways from local sandy soil to groundwater and from the soil to plant were examined
by lysimeter studies and greenhouse pot experiments. The fate of the comparable large and heavy X-ray contrast media DZA
and the antiepileptic CBZ as common indicators for OMP was analyzed. This should allow for a first risk estimation based on
realistic OMP concentrations in the RW.
DZA showed tracer-like transport in the lysimeters without retardation posing a high risk for groundwater contamination.
Also, DZA was found to accumulate in or on the roots of arugula with a remarkable RCF of 1,925 + 34% in pot experiments.
Probably it is transported with the water flux due to its high mobility but is not able to pass through the epidermis due to its
size and negative charge. However, a small part of DZA was quantified in arugula leaves with a BCF of 18 + 22%.
CBZ seems to be removed in the very upper soil layers in the lysimeter studies. It, therefore, poses no risk for groundwater
contamination for the sandy soil of our case study. However, other studies showed vertical CBZ transfer in sandy soil that
potentially indicates a long-term risk also for groundwater in Brandenburg. Contradicting, CBZ could not be found in greater
concentrations in the soil material of the pot experiments demonstrating (a) no or barely any sorption to it or (b) probably
transformation. However, CBZ showed an uptake by arugula with 70 + 7% RCF and 155 + 12% BCF indicating a potential
transfer toward plants.
While bringing together these perspectives, water reuse with RW can be a promising option for agricultural irrigation in
Brandenburg, if water quality is managed with regard to OMP and potential transfer pathways. The two examples, DZA

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 Water Reuse Vol 14 No 1, 12

and CBZ, showed a wide range of possible environmental fates and related risks due to substance-specific properties. Com-
parison with literature data showed that soil properties (inorganic, organic, and microbiological) together with cultivated
plants alter OMPs’ fate.
For a sound risk management, fate predicting tools for wastewater treatment, the pathway soil to groundwater, and the
pathway soil to plant would be very helpful. As this has to be based on intrinsic molecule properties, local soil properties,
and agricultural product, this is a major challenge, especially since there is a lack of general parameters (e.g. DOW or
DOC) to predict sorption of potentially ionic OMPs (Sigmund et al. 2022).
However, the load of OMPs in RW is the driving parameter for concentrations found in the environment. It is, therefore,
highly recommended to implement advanced wastewater treatment (tertiary treatment). In addition, treatment should be
selected based on the case-specific OMPs, which are considered most likely to contaminate groundwater and plants in the
area where the water is to be reused.
As shown for DZA and CBZ, polar and mobile as well as non-polar and non-mobile OMPs are transferred into agricultural
products. Hence, to address all these aspects, monitoring and extended analyses of OMPs are needed to minimize potential
risks.

ACKNOWLEDGEMENTS
We thank Silke Pabst for HPLC-MS/MS analyses and Pia Schünemann for sample preparation and peak integration for SFC-
MS/MS analyses. We are very grateful to Manuela Helmecke for her critical proofreading of the final manuscript. This inves-
tigation was supported by the German Federal Ministry of Education and Research (BMBF) within the project PU2R (UBA
2021), contract 02WV1564.

DATA AVAILABILITY STATEMENT

All relevant data are included in the paper or its Supplementary Information.

CONFLICT OF INTEREST
The authors declare there is no conflict.

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