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water research 47 (2013) 2661 e 2675 Available online at www.sciencedirect.com journal homepage:

Available online at www.sciencedirect.com

Available online at www.sciencedirect.com journal homepage: www.elsevier.com/locate/watres

journal homepage: www.elsevier.com/locate/watres

journal homepage: www.elsevier.com/locate/watres The use of d 1 5 N and d 1 8 O tracers
journal homepage: www.elsevier.com/locate/watres The use of d 1 5 N and d 1 8 O tracers

The use of d 15 N and d 18 O tracers with an understanding of groundwater flow dynamics for evaluating the origins and attenuation mechanisms of nitrate pollution

Takahiro Hosono a , * , Takahiro Tokunaga b , Makoto Kagabu b , Haruhiko Nakata b , Takanori Orishikida b , In-Tian Lin b , Jun Shimada b

a Priority Organization for Innovation and Excellence, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555, Japan b Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555, Japan

article info

abstract

Article history:

During early 2000, a new analytical procedure for nitrate isotopic measurement, termed

Received 7 September 2012

the “denitrifier method”, was established. With the development of the nitrate isotope

Received in revised form

tracer method, much research has been reported detailing sources of groundwater nitrate

4 February 2013

and denitrification mechanisms. However, a shortcoming of these tracer studies has been

Accepted 7 February 2013

indicated owing to some overlapping of isotope compositions among different source

Available online 19 February 2013

materials and denitrification trends. In order to reduce these uncertainties, we examined

nitrate isotope ratios within a frame of “regional groundwater flow dynamics” to eliminate

 

Keywords:

unnecessary uncertainties in elucidating nitrate sources and behaviors. A total of 361

Groundwater

samples were collected from the Kumamoto area: the circulated groundwater system with

Flow dynamics

a scale of 10 3 km 2 in southern Japan. Subsequently, the nitrate pollution was examined

Nitrate pollution

within the above-mentioned framework. As a result, a reasonable identification of the

Stable isotope ratios

sources and attenuation behaviors (both denitrification and dilution) of groundwater ni-

Denitrification

trate pollution was obtained over the study area. This study demonstrates that the use of

nitrate isotope tracers efficiently improves with a comprehensive understanding of

groundwater flow dynamics. The approach emphasized in this study is important and

should be applicable in other areas.

ª 2013 Elsevier Ltd. All rights reserved.

1.

Introduction

Groundwater nitrate pollution is one of the most prevalent and on-going global natural water environmental problems ( Rivett et al., 2008). The comprehension of nitrate pollution sources and natural attenuation mechanisms is of prime importance for better management of groundwater resources. The stable isotope ratio of nitrogen in nitrate ( d 15 N NO 3 ) has been used since the 1970s as a powerful tracer revealing concealed pollution sources and to understand the mecha- nisms of denitrification (i.e. Kreitler, 1979; Kaplan and

Magaritz, 1986; Mariotti et al., 1988; Smith et al., 1991). By the early 1990s, the combined use of d 15 N NO 3 with the stable isotope of oxygen in nitrate ðd 18 O NO 3 Þ was proposed and applied for groundwater study (for example Bo¨ ttcher et al., 1990; Wassenaar, 1995; Aravena and Robertson, 1998; Fukada et al., 2003). In addition, the newly established analytical procedure of nitrate isotopic measurement termed the “denitrifier method” (Sigman et al., 2001; Casciotti et al., 2002)

enables the determination of dual d 15 N NO 3 and d 18 O NO 3 using a

3 concentra-

smaller volume of water (<8 ml) with lower NO

tion in quicker preparation time than previous methods. The

* Corresponding author . Tel./fax: þ 81 96 342 3935. E-mail address: hosono@kumamoto-u.ac.jp (T. Hosono). 0043-1354/$ e see front matter ª 2013 Elsevier Ltd. All rights reserved.

nitrate isotope tracer method has become established as a better tracing tool and has increasingly been applied in many parts of the world (i.e. Umezawa et al., 2008; Otero et al., 2009; Hosono et al., 2011a). Within a nitrate isotope tracing study, the pollution sour- ces can be estimated by isotopic comparison between the water samples and possible source materials (Kendall, 1998). The isotopic comparison can also be used as a denitrification indicator by analyzing isotopic enrichment trends of water samples ( Kendall, 1998). However, source distinguishment becomes difficult when different possible source materials, such as manure and sewage, have a similar isotope signature

(Xue et al., 2009; Fenech et al., 2012; Minet et al., 2012). More- over, if isotopic compositions overlap between the source characteristics and denitrification trends, this distinction be- comes difficult ( Xue et al., 2009). These difficulties complicate data interpretation. Therefore, it has recently been argued that a new approach and concept are needed to reduce the above mentioned shortcomings in nitrate isotope tracer studies ( Xue et al., 2009; Fenech et al., 2012; Minet et al., 2012). In a circulated groundwater system, dissolved nitrate is introduced to groundwater at the recharge zone and trans- ported through the lateral flow zone to the discharge zone within a frame of “groundwater flow dynamics” (Bo¨ hlke and Denver, 1995; Bo¨ hlke et al., 2002; Hosono et al., 2011b). Therefore, having a good understanding of the three- dimensional groundwater flow pathway is very important when tracing complicated nitrate sources and behaviors using a nitrate isotope tracer. In addition, it is important to prepare detailed land-use information for the target area, especially for the assessment of potential impacts from manure and sewage (Xue et al., 2009; Fenech et al., 2012; Minet et al., 2012). The nitrate isotope tracer study method with a combined use of the above mentioned control information appears to be an efficient approach for reducing uncertainties in the past study. However, emphasis has not been clearly placed on its importance in previous articles. Kumamoto City is located in the central part of the Kyushu Islands in southern Japan. The city is known as the largest groundwater utilization region in Japan where about one million people in and around the city depend entirely on groundwater for drinking purpose ( Oshima, 2010; Shimada, 2012). However, the monitoring data for groundwater quality show that nitrate concentrations have been increasing since 1970s (Tomiie et al., 2009, 2011). For example, NO 3 concen- trations for some wells ( Fig. 1a) increased by a factor of 2 e7 during the last 40 years ( Appendix 1 ). Although nitrate source controls have been implemented since the 1990s, such as control of fertilizer applications and the construction of bar-

riers around manure storage sites, groundwater NO

3 is still

increasing and becoming an increasingly serious environ- mental issue (Kumamoto Prefecture and Kumamoto City, 2005; Kumamoto City Waterworks and Sewerage Bureau, 2008; Kumamoto City, 2009; Oshima, 2010). However, the principal causes of the pollution and natural attenuation mechanisms have not been fully understood. This study presents a comprehensive amount of nitrate isotope data (total of 361 samples) for groundwater (confined and unconfined) and comparison water (spring and river water) during different seasons, and possible source materials

(chemical fertilizers, manures, and sewage water) taken from the Kumamoto area. These data were used within a tracer study in the context of groundwater flow dynamics. We firstly tried to understand the general view of the origins of ground- water nitrate pollution, and then examined detailed attenua- tion mechanisms along the flow paths by denitrification as well as by mixing of the different source water. The results from these investigations were used to emphasis the useful- ness of the above approach for reducing uncertainties encountered in previous studies, which is the prime objective of this study. The approach could be applicable in future studies for a better understanding of the sources and attenu- ation mechanisms of groundwater nitrate pollution.

2. Outline of the research area

2.1. Topography and climate

The Kumamoto groundwater area is defined as an area (945 km 2 ) surrounded by the divide of the Shira River water- shed to the north, the Midori River to the south, the Aso Mountain (1592 m) to the east, and the Ariake Sea and Kinpo Mountain (665 m) to the west ( Fig. 1 a). The altitude of this area tends to decrease westward from land to ocean. Importantly, volcano-pyroclastic and lava flow deposits derived at the time of the Aso Caldera formation formed three major highlands with altitudes around 100 m: the Kikuchi and Takayubaru highlands extending westwards and the Ueki highlands to the north of the Kumamoto area ( Fig. 1a). In addition, volcanic flow deposits of the Pre-Aso activity formed small hills in the central area such as Tatsuta Mountain (Fig. 1a). Kumamoto had an average annual temperature and precipitation of 16.9 C and 1986 mm, respectively during 1980e2010. The rainy season occurs from JuneeJuly with around 40% of the total annual precipitation occurring during this period.

2.2. Geology

The study area is composed of four major geological constit- uents (Fig. 2 ): the Paleozoic basement of metamorphic and sedimentary rocks; the Pre-Aso volcanic rocks of Tertiar- y eQuaternary age; the Quaternary Aso volcanic rocks; and overlaying alluvium deposits. Paleozoic basement rocks do not outcrop at the surface within the study area. The Pre-Aso volcanic rocks generally consist of lavas and tuff breccia ( Ono and Watanabe, 1985; Miyoshi et al., 2009), while the Aso vol- canic rocks are composed of pyroclastic deposits and flow lavas ( Ono and Watanabe, 1985). The former rocks are present

at/around the Kinpo and Tatsuta Mountains ( Fig. 1 a), while latter rocks are distributed around the eastern side of the Kinpo Mountain and the western side of the Aso Caldera. Both rock types have a wide chemical range between basalticer- hyolitic compositions ( Ono and Watanabe, 1985; Miyoshi et al., 2009). The Aso volcanic rocks are subdivided into four units according to eruption stages: Aso-1 (270 ka); Aso-2 (140 ka); Aso-3 (120 ka); and Aso-4 (89 ka) ( Ono and Watanabe, 1985). Of these, the Aso-2 unit is unique in its composition of andesitic lava with joints and porous structures. It is also important to note that clay impermeable layers of lacustrine

water research 47 (2013) 2661 e 2675 2663 Fig. 1 e A map showing (a) topography

Fig. 1 e A map showing (a) topography and major groundwater flows and (b) land-use and sampling locations in the Kumamoto area. Solid contour lines with numbers in Fig. 1 a represent groundwater levels (m, above sea level) calculated with a groundwater flow model using a surface and subsurface coupling flow simulator (GETFLOWS: Tosaka et al., 2000). Details for groundwater flow model will be shown in separated paper.

sediments are widely distributed between the Aso-3 and Aso- 4 units (Fig. 2 ). The other clay layer of marine origin, the Ariake clay layer, is distributed over the Kumamoto plain especially within the coastal zone.

2.3.

Hydrogeology

The highly permeable structure of Aso-4 pyroclastic deposits at the Ueki, Kikuchi, and Takayubaru highlands result in these

areas becoming major groundwater recharge sources within Kumamoto ( Kumamoto Prefecture and Kumamoto City, 1995; Taniguchi et al., 2003). From a hydrogeological point of view, the Aso volcanic rocks and alluvial sedimentary deposits are the major aquifers which overlie impermeable Paleozoic basement and Pre-Aso volcanic rocks (Kumamoto Prefecture and Kumamoto City, 1995) ( Fig. 2). The aquifers are divided into two main types: unconfined and confined aquifers ( Fig. 2). The former aquifer type is composed of unwelded Aso-4

2664 water research 47 (2013) 2661 e 2675 Fig. 2 e Hydrogeological cross sections between A

Fig. 2 e Hydrogeological cross sections between AeA 0 and B eB 0 in Fig. 1. Arrow in the figures represents groundwater flow direction at each grid obtained by the results of a groundwater flow model calculation using a surface and subsurface coupling flow simulator (GETFLOWS: Tosaka et al., 2000). Size of arrow corresponds to relative velocity of groundwater flow. Values for estimated enrichment factor ε are also shown in figures for each groundwater flow zone (see text for details).

pyroclastic and alluvial sedimentary deposits, which overlie an aquiclude of impermeable lacustrine sediment layer. The latter aquifer type consists of Aso-2 porous andesitic lava and pyroclastic deposits of Aso-1, Aso-2, and Aso-3 units. As an important consideration, the aquiclude is not continuously distributed under the mid-stream area of the Shira River (Figs. 1a and 2 ). Due to the very high downward infiltration capacity of this area ( w500 mm d 1 ), a large amount of water is recharged from the river into the deep aquifer ( Tanaka et al., 2010) ( Fig. 2 ). Groundwater is situated generally at w50 m under the surface for unconfined and around 60e 200 m for confined aquifers. The groundwater levels for both aquifers show 5 e10 m seasonal change during a year. In general, the high water season is from AugusteNovember and low water season is from JanuaryeApril.

2.4. Groundwater flow and age

Groundwater flow systems are well understood within the study area based on long-term water level monitoring and

groundwater modeling (Figs. 1a and 2). The most significant flow system of confined groundwater is the flow A eA 0 in Figs. 1 a and 2. This main flow can be classified into 4 zones (Figs. 1 a and 2): A1- the flow from the recharge area of the Kikuchi highland to the mid-stream area of the Shira River; A2- the mixing zone of groundwater from zone A1, recharged water from the Takayubaru highlands and the southwestern feet of Aso Mountain, and the recharged water from the Shira River; A3 e the southwestern lateral flow through the Aso-2 volcanic lava aquifer toward the discharge area of the Ezu Lake with a discharge rate of 1.5 10 8 m 3 y 1 (Shimada, 2012); and A4 e the flow from the discharge area to the coastal stagnant area. The flow Be B 0 is the other important confined groundwater flow system within the study area (Figs. 1a and 2), which can be classified into 3 zones along the flow direction: B1 e southward flow from the recharge area of the Ueki highlands to the point of basement lifting; B2 e the flow across the basement uplifting toward the entrance of plain area; and B3 e southwestward flow to the coastal stagnant area. The number of themonitoring wells for the unconfined groundwater is limited compared to

water research 47 (2013) 2661 e 2675 2665 Fig. 3 e Comparison of NO groundwater, spring

Fig. 3 e Comparison of NO

groundwater, spring water, and river water samples from the Kumamoto area.

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concentrations among

that of confined groundwater. However, the general view of the flow character is similar to each other, which is basically controlled by regional geology and topography. According to unpublished 3 H, chlorofluorocarbon, SF 6 , and 85 Kr age data (Kagabu et al., personal communication), the groundwater ages range from few years at the recharge zone to 25e30 years at the discharge zone. The stagnant groundwater under the coastal plain (zone A4 and B3) is thought to be old groundwater with an age more than 50 years.

2.5. Land-use and groundwater use

Land-use in the study area is shown in Fig. 1b. The western mountain foot of the Aso Mountain is widely covered by forest and grassland, with anthropogenic loading being relatively small in this area. Vegetable farming is the main land-use in the Ueki, Kukuchi, and Takayubaru highlands, which has been present since the early 1900s. In addition, livestock farming has increased in this area since the 1970s. For example, the numbers of livestock (cows and pigs) increased by a factor of about 30 during the last 40 years ( Tomiie et al., 2011). On the contrary, paddy is distributed at lowland areas along rivers and plain area (Fig. 1).

The study area has population of about one million. People in the area depend entirely on confined groundwater for do- mestic water resources including drinking purpose ( Oshima, 2010). A total of 8 10 7 m 3 y 1 of groundwater is pumped

from 58 pumping stations installed within the study area and distributed to each house. The percentage of population with access to municipal sewage treatment plant in the city is about 86%. Treated water at several sewage treatment plants

is released directly to nearby rivers.

3. Sampling and methods

The sampling locations are shown in Fig. 1b. A total of 239 groundwater samples (83 for unconfined and 156 for confined groundwater) were collected during November 2009e November 2011. To account for the high water season, groundwater sam- ples were collected during November 2009, October 2010, and OctobereNovember 2011, respectively. Representing the low water season, samples were collected during JanuaryeMarch 2011 at the same wells as surveyed during the high water season in OctobereNovember 2011. A total of 86 river and spring water samples were also collected during April 2011 (low water sea- son) and July 2011 (high water season), respectively. Further- more, 16 sewage effluent samples for both pre-treated and treated water from five representative sewage treatment plants, ten chemically synthesized NPK fertilizer samples, and ten manure samples were obtained for possible source materials. Collected fertilizers are the most representative fertilizers generally used over the study area for fruits, vegetables, and rice. Manure samples were collected in the livestock farms at Ueki, Kikuchi, and Takayubaru highlands.

Groundwater samples were collected using an internal or

portable pump (MP1, Grundfos). Prior to sampling of borehole water, old water in the pipes (w5 10 3 l; the volume was calcu- lated based on borehole depth and casing size) was pumped out to collect fresh water as much as possible. Parameters such as temperature, electrical conductivity (EC), pH, oxidation reduction potential (ORP), and dissolved oxygen (DO) were continuously monitored during pumping using fieldmeters (HORIBA D-54) and water samples were collected after these values became con- stant. Collected water samples for analysis of nitrate concen-

trations and nitrate isotope ratios were filtered through 0.2 mm cellulose-acetate filters before storing in 50 ml volume poly- ethylene bottles. Filtered samples were kept immediately in ice box after sampling, transported to laboratory within a day, and stored in freezer with a constant controlled temperature of 15 C until just before analysis. Manures and chemical fertil- izers were freeze dried and pulverized using an agate mortar. NO concentrations were measured by ion chromatography (Compact IC 761, Metrohm, Switzerland). The d 15 N NO 3 and d 18 O NO 3 compositions were determined following the “denitrifier method” of Sigman et al. (2001) and Casciotti et al. (2002). Briefly,

sample containing NO was converted to nitrous oxide (N 2 O) by denitrifying bacteria that lack N 2 O-reductase activity. The

a

3

3

N 2 O was stripped from the sample vial using helium carrier gas,

purified using cryogenic trapping (Thermo Fisher Precon Sys- tem), chromatographically separated (Thermo Fisher Gas Bench), and analyzed using mass spectrometry (Thermo Fisher Delta V Advantage). The ratios of 15 N: 14 N and 18 O: 16 O were

2666 water research 47 (2013) 2661 e 2675 Fig. 4 e Seasonal comparison between the high

Fig. 4 e Seasonal comparison between the high and low water season for (a) NO , (b) d 15 N NO 3 , (c) d 18 O NO 3 , and (d) dual d 15 N NO 3 and d 18 O NO 3 compositions. Two solid lines in Fig. 4 d represent isotopic fractionation trends during denitrification with d 15 N NO 3 :d 18 O NO 3 of 1.3:1 and 2.1:1, respectively.

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expressed relative to atmospheric nitrogen for nitrogen and Vienna Standard Mean Ocean Water (V-SMOW) for oxygen. Isotope valueswere calibrated using the United States Geological Society (USGS) and International Atomic Energy Agency (IAEA) internationally recognized nitrate standards: USGS-34, USGS-35, and IAEA-N3. Based on replicate measurements of standards and samples (n ¼ 288), the analytical precisions for d 15 N NO 3 and d 18 O NO 3 were better than 0.2& and 0.3&, respectively. The nitrogen isotope ratio for powdered samples was determined by mass spectrometry interfaced with a CNS elemental analyzer (Thermo Fisher Flash 2000). Analytical precision of the analysis was better than 0.15& .

4.

Results

4.1. Nitrate concentrations

The groundwater NO concentrations ranged between 0 and 73 mg l 1 and only 3 samples among the total analyzed

3

samples ( n ¼ 239) exceeded the World Health Organization limit of 50 mg l 1 . Therefore, current nitrate concentration level was judged to be generally lower than environmental criteria, although increasing trend of concentrations is a threat to its drinking water supply in future for many bore-

hole sites ( Tomiie et al., 2009 , 2011 ) ( Appendix 1 ). Similar frequency patterns were observed for nitrate concentrations between confined and unconfined groundwaters ( Fig. 3 ). The

groundwater tended to be more NO

3 enriched than river

water but showed a similar concentration range to spring

water ( Fig. 3 ). NO

3 concentrations basically were not

changed between water samples collected at high and low

3 concentrations for

water seasons ( Fig. 4 a). The highest NO

both confined and unconfined groundwater samples were found in the recharge areas at the Ueki, Kikuchi, and Takayubaru highlands where vegetable and livestock farms

are highly concentrated ( Fig. 5 a). It was also observed that

NO

3 concentrations clearly decreased along the ground-

water flow path from recharge edischarge/stagnant zones

( Fig. 5 a).

4.2. Nitrate isotope ratios

The d 15 N NO 3 and d 18 O NO 3 of groundwater samples ranged be- tween 6e46& and 3e48&, respectively ( Fig. 6). An isotopic compositional gap was not found between confined and un- confined groundwaters ( Fig. 6). Groundwater samples with moderate to high NO concentrations tended to have narrow isotope compositions (d 15 N NO 3 ¼ 2e8 & and d 18 O NO 3 ¼ 3e5 &) ( Fig. 6), which are similar to the compositional ranges of spring water samples. In contrast, the samples with the

3 concentrations tended to have higher isotopic

lowest NO

compositions ( d 15 N NO 3 ¼ 8e46& and d 18 O NO 3 ¼ 5 e48&), sug- gesting the occurrence of denitrification. The spring and river water d 18 O NO 3 compositions were similar to each other, whereas d 15 N NO 3 compositions of river water were clearly higher than those of spring water ( Fig. 6 ). Seasonal gaps of d 15 N NO 3 and d 18 O NO 3 were not observed for all water samples, except groundwater samples showing highly fractionated signatures ( Fig. 4b and c). These ground- water samples were unique in that the fractionation ratios of d 18 O NO 3 to d 15 N NO 3 were highly variable ( Fig. 4 d). It has been commonly thought that the fractionation ratio of

d 15 N NO 3 :d 18 O NO 3 during denitrification ranges within 1.3e2.1:1 ( Aravena and Robertson, 1998; Mengis et al., 1999; Bo¨ ttcher et al., 1990; Fukada et al., 2003). However, some groundwater samples collected in our study showed more variable ratios than the reported range ( Figs. 4 and 6 ). In fact, the cause of variability of fractionation ratios is becoming recognized as a matter of discussion (Kool et al., 2011), which mechanism has not been solved yet. This will not be discussed in this paper; however, results of this study suggest that d 15 N NO 3 and d 18 O NO 3 values of groundwater should be regarded as subject of vari- ation during denitrification over different seasons. The calculation of the average value is one of the approaches of data treatment as shown in this study. The nitrogen isotope ratios of the possible source materials showed their own signatures with some compositional vari- ations ( Fig. 6). Remarkable isotopic compositional changes within collected fertilizer samples were not found among different production companies, type of farm products used for, and sampling areas. Similarly, isotope compositions of manure samples did not change depending on their sampling areas. Moreover, no isotopic changing tendencies were observed for sewage waters collected at different conditions. It could be considered from these observations that nitrogen isotope variation for each source material regards as isotopic heterogeneity of material itself, but is not due to the factors of space and probably of time. The compositional range of d 15 N for chemical fertilizers was similar to that of groundwater samples showing high

NO

3 concentrations ( >30 mg l 1 ), whereas sewage and

manure samples showed higher values ( Fig. 6 ). The

groundwater samples tended to increase in d 15 N NO 3 (and

d 18 O NO 3 ) with decreasing of NO

along the flow direction from recharge edischarge/stagnant zones ( Fig. 5 ). The pollutant input locations of manure with high d 15 N NO 3 should be in/around the recharge highlands area. The results indicated that increasing groundwater

d 15 N NO 3 in the plain and coastal areas is not due to the contribution of manures.

3

3 and DO concentrations

to the contribution of manures. 3 3 and DO concentrations Fig. 5 e Distribution map for

Fig. 5 e Distribution map for (a) NO , (b) d 15 N NO 3 , and (c) DO in the study area. Arrows indicate the major flow directions of the confined groundwater.

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2668 water research 47 (2013) 2661 e 2675 Fig. 6 e Nitrate isotope comparison for water

Fig. 6 e Nitrate isotope comparison for water samples with possible source references. Note that the data shown with an asterisk are taken from Hirohata et al. (1999).

5.

Discussion

5.1. Source of nitrate pollution in groundwater

It is clear from the observed nitrate distributions ( Fig. 5a) that groundwater nitrate for both confined and unconfined

3 concentrations >30 mg l 1 was

transported through the recharge area where vegetable and livestock farms are widely distributed. This groundwater is characterized by DO concentration values > 6 mg l 1 ( Fig. 5 c). Under such conditions, it is commonly considered that deni- trification would not preferentially occur ( Spalding and Exner, 1993; Cey et al., 1999; McCallum et al., 2008). Importantly, such high NO groundwater ( >30 mg l 1 ) generally has d 15 N NO 3 and d 18 O NO 3 compositions within 2e8 & and -3 e5& , respectively with no significant isotopic fractionation between them (Fig. 6 ). Therefore, it can be roughly estimated that the effects of denitrification were not significant. From an isotopic com- parison ( Fig. 6 ), it is most likely that concentrated nitrate of groundwater as well as those of spring water (where DO concentration was in general >8 mg l 1 ) was dominantly originated from long-term application of chemical fertilizers

groundwater with NO

3

to the vegetable farms. However, with the fact that these high nitrate water samples showed some d 15 N NO 3 variation (2e 8&) partial contributions of manures from livestock farms and organic fertilizers cannot be neglected. One confined groundwater sample at the recharge area

3 (31 mg l 1 ) content

( Fig. 6). This isotope composition corresponds with that of manure. However, from its elevated d 18 O NO 3 (8.7&) value and depleted DO concentration value (1.3 mg l 1 ), it can be sug- gested that the sample properties were due to the denitrifi- cation of the nitrate that originated from chemical fertilizers under a sporadically anaerobic environment. The number of houses is limited within the farming area. Even if some houses are not connected to the sewer, domestic effluents generally drain to nearby rivers through side ditches. Leakage from sewage pipes is another possible trans- portation source of domestic wastes in urbanized areas ( Lerner, 2002; Wakida and Lerner, 2005). In Kumamoto, the downward flowing groundwater from the recharge area generally did not display clear tendency of NO concentration increase with an increase of d 15 N NO 3 in urbanized areas ( Fig. 5 ). Therefore, it is likely that the contribution of sewage leaked from sewage pipes is not the prime source of nitrate, although

showed a high d 15 N NO 3 (14.9&) and NO

3

water research 47 (2013) 2661 e 2675 2669 Fig. 7 e Changes of NO , DO,

Fig. 7 e Changes of NO , DO, d 15 N NO 3 , and d 18 O NO 3 with groundwater flow (a) A eA 0 and (b) B eB 0 . Plots in Fig. 7 correspond to groundwater samples collected at borehole sites within the solid lines in Fig. 1. The parallel axis indicates sampling points in order of groundwater flow direction for A eA 0 and BeB 0 , respectively. The order was decided in Fig. 1 from the connecting positions between the lines A eA 0 (or BeB 0 ) and the perpendicular line from each site. Compositional variation in each site represents seasonal variation.

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some contribution of it is suggested by the analysis of chem- ical markers (artificial sweeteners such as acesulfame K) (Nakata et al., personal communication), which is a more sensible tracer of sewage source, and by the analysis with flow systems (discussed later). Treated wastewater from sewage treatment plants, which still show high NO concentrations in the order of few tens of mg l 1 , drain directly into nearby rivers. As mentioned previously, river water (with DO con- centrations >8 mg l 1 ) showed higher d 15 N NO 3 compositions than do spring water and groundwater with high NO concentrations > 30 mg l 1 ( Fig. 6). It is reasonable to conclude that nitrate in river water was partly derived from wastewater of sewage origin. As a consequence, it can be concluded that long-term application of chemical fertilizers is a prime cause of

3

3

groundwater nitrate pollution presently observed in Kuma- moto. Minor contributions of manures from livestock farms and organic fertilizers might also be suggested as secondary important sources. It is however difficult to point out their contribution rates for each sampling site due to the fact that

isotopic compositions of the source materials have their own variations. Finer scale mixing signature of these sources would be better expressed on a simulation model by putting information of historical change of the land-use and nitrate

loads on the groundwater flow simulation model and by tuning between calculated results and observed concentra- tion and isotopic data. We are preparing to show these results in a separated paper. Our results taken in context with this study imply that the effects of political action in the control- ling of fertilizer applications enforced after the 1990s are not

2670 water research 47 (2013) 2661 e 2675 Fig. 8 e Sample plots on the d

Fig. 8 e Sample plots on the d 15 N NO 3 vs. NO

dilution and (b) denitrification, and (c) for the Midori River watershed named as zone C groundwater. Plots for dilution type groundwater samples are also shown in Fig. 8b in a yellow color. (For interpretation of the references to colour in this figure

legend, the reader is referred to the web version of this article.)

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diagram for groundwater with two different nitrate attenuation types: (a)

yet evident. Groundwater nitrate monitoring along with d 15 N NO 3 and d 18 O NO 3 analysis is important not only to check pollution status but also to evaluate the effects of manure, which might be the next additional concern as a pollution source.

5.2. Two different attenuation mechanisms: dilution

and denitrification

3 concentration and

d 15 N NO 3 changes were observed along the flow direction be- tween the two different groundwater flow systems: A eA 0 and B eB 0 ( Fig. 7). This implies that different attenuation processes for nitrate pollution are occurring within each flow system. This section describes the attenuation mechanisms for each flow system in detail and compares their efficiencies in terms of attenuation functions.

The contrasting decline tendency in NO

5.2.1. Groundwater flow AeA 0

In zone A1, nitrate concentration found in confined ground- water decreased along the flow direction ( Fig. 7 a). However, the DO concentration values were still higher than 8 mg l 1 and the d 15 N NO 3 change was very small (1e1.5 &). Moreover,

isotopic fractionation was also unclear for d 18 O NO 3 ( Fig. 7 a). In turn, it is well known that paddy irrigation water of the Shira

River water (NO

DO ¼ 8e9 mg l 1 ) recharges underneath groundwater at the

mid-stream area of the Shira River ( Tanaka et al., 2010) ( Fig. 2).

3 concentration in

groundwater decreased along the flow direction due to dispersion and/or dilution by irrigation water, but not by denitrification. Water quality within the zone A2 largely var- ied ( Fig. 7a), mainly due to the zone A2 being a mixing zone

among several different types of water. However, the d 15 N NO 3 compositions were homogeneous (1.5e5.5&) with the range almost equivalent to that of zone A1 (3.4e5.0 &) ( Fig. 7a). Observed heterogeneity in NO concentrations should not be due to denitrification but to the mixing of different water of varying NO concentrations.

3 concentration in confined ground-

water gently decreased from 20 to 15 mg l 1 with a small in-

crease of d 15 N NO 3 ( w6& ) through zone A3 flow ( Fig. 7a). The DO concentration values of groundwater averaged around 8 mg l 1 throughout the zone, supporting the idea that deni- trification is not a controlling factor in the decline of nitrate concentrations. Nagai et al. (1983) revealed from water chemistry data that confined groundwater in this zone is the mixture of up-gradient groundwater and Shira River water with a proportion of about 3e 4:6e7. Considering our results in the context of this fact, it can be concluded that the observed decline in NO concentrations and the increase in d 15 N NO 3 are attributed to dilution by Shira River water of relatively lower

NO

3 concentration and higher d 15 N NO 3 value (average of

5.5 mg l 1 and 7.9&, respectively; n ¼ 2), which is recharged at the mid-stream area of this river. In Fig. 7a, water chemistry and nitrate isotopic profiles from zone A1eA3 appear similar between confined and unconfined groundwater, suggesting that similar processes explained above were also taking place for unconfined groundwater.

3 ¼ 5e 6 mg l 1 , d 15 N NO 3 ¼ 7e10&,

These observations suggest that the NO

3

3

It is notable that NO

3

Toward the end of zone A4, confined groundwater samples

3 concentration with a decreasing

displayed a decline of NO

of DO concentration ( Fig. 7a). However, d 15 N NO 3 fractionation

was not observed and clear evidence of denitrification was not confirmed. Therefore, this might be interpreted to be owing to the weakening of pollution influence to the deep part of stagnant groundwater. In contrast, unconfined

groundwater samples showed an increasing trend of d 15 N NO 3

3 concentrations along

with the decreasing of DO and NO

the flow direction (Fig. 7a). It was the only evidence of the

occurrence of denitrification within the groundwater flow system AeA 0 .

5.2.2. Groundwater flow Be B 0

A clear decline of NO concentrations (50e1 mg l 1 ) with the flow direction was confirmed for zone B1 confined ground-

water accompanied with a major decrease of DO concentra- tion values (8e1 mg l 1 ) and the increasing of d 15 N NO 3 values (4e8 &) (Fig. 7 b). This indicates the progress of the denitrifi- cation reaction ( Knowles, 1982; Hiscock et al., 1991; Korom, 1992; Mariotti, 1986). At the entrance of zone 2B over the

3 and DO concentra-

tions increased to 20 mg l 1 and 7 mg l 1 , respectively, while

the d 15 N NO 3 decreased to 3& ( Fig. 7b). It appears likely that water with dissolved materials could be transported through the thinning part of the aquifer and the discontinuity of the aquiclude (Fig. 2). Subsequently, through the flow toward

3 declined again

(20e 1.5 mg l 1 ) with a very sudden depression of DO

(7e1 mg l 1 ) and with a major increase of d 15 N NO 3 (up to 40& in

maximum) ( Fig. 7b), showing clear evidence for the drastic progress of the denitrification reaction. This zone is situated under a topographical changing point where the terrestrial hills meet with plain area covered by marine and flood sedi-

ments (Figs. 1a and 2). Considering that marine clays and flood deposits are more enriched in organic materials than igneous rocks and with its low DO feature, this area is regarded as a

denitrification hotspot similar to a riparian zone where sig- nificant denitrification preferentially occurs ( Bo¨ hlke and Denver, 1995; Bo¨ hlke et al., 2002; Hoffmann et al., 2006). This

area is termed a “hills-plain border zone”. After water passes this zone, the d 15 N NO 3 composition returns again to low values between 0 and 10 & at the entrance of zone 3B ( Fig. 7 b). This indicates the scenario that the ma- jority of nitrate from zone 1Be 2B has been consumed by denitrification and has not reached this point. The NO 3 con- centration was low under the Shira River area (around 1 mg l 1 ). Naturally occurring nitrate in the aquifer geology might be the origin of this modestly concentrated nitrate. In

3 concentrations in confined groundwater

zone 3B, NO

decreased ( w0 mg l 1 ) with a significant increase of d 15 N NO 3 up

to 30& ( Fig. 7b). Groundwater in this zone is in addition characterized by low DO concentration values generally <2 mg l 1 , suggesting the occurrence of denitrification. A small influence of nitrate infiltration from the unsaturated zone was confirmed in zone 3B; however, the general ten- dency of changing patterns of NO , DO, and d 15 N NO 3 for un- confined groundwater was quite similar to that of confined groundwater throughout the flow distance ( Fig. 7 b). Therefore, similar processes should be affecting both groundwater types.

3

basement uplifting point, the water NO

beneath the Shira River, groundwater NO

3

2672 water research 47 (2013) 2661 e 2675 Fig. 9 e Estimation of the isotopic enrichment

Fig. 9 e Estimation of the isotopic enrichment factor ε for each zone.

5.2.3. Efficiencies of the two different attenuation functions

Fig. 8 compares the effects of the two different attenuation types on groundwater nitrate pollution judged in the previ-

ous discussion: dilution ( Fig. 8 a) and denitrification ( Fig. 8 b). Groundwater for both attenuation types showed highest

concentrations around 50 mg l 1 , which were largely

derived from chemical fertilizers. The NO

decreased to 20 e10 mg l 1 for most of the samples attenu- ated by the dilution with lower nitrate water ( Fig. 8 a). From the plots distribution in Fig. 8 a and previous discussion, it could be suggested that the lower nitrate water is the mixture of the least contaminated (or non-contaminated) groundwaters and the recharged river waters moderately

3 concentrations

NO

3

contaminated by sewage origin nitrate. On the other hand,

3 concentrations decreased to <1 mg l 1 by the action of

denitrification ( Fig. 8 b). This means that about 60 e80% and

98% of the nitrate load was naturally purified by dilution and

denitrification, respectively. It is clear from this comparison that denitrification is more efficient in terms of the nitrate attenuation function for circulated groundwater systems.

3 attenuation is there-

Groundwater affected by dilution NO

fore, more vulnerable to the threat of additional pollution

such as in zone A3 (Figs. 1 a and 8 ) especially at where dilution water has already been contaminated in some extent. In a closed system, isotopic enrichment during denitri- fication can be expressed by the following Rayleigh equation:

NO

d 15 N NO 3 ¼ d 15 N NO 3 i þ εlnð C=C i Þ

(1)

where ε is the isotopic enrichment factor (a parameter for denitrification activity), d 15 N NO 3i and d 15 N NO 3 represent isotope compositions of water at the initial time and sample time, respectively, and C i and C are the concentrations of nitrate in water at the initial time and sample time, respectively. Equation (1) can be used to estimate the enrichment factor ε by plotting measured d 15 N NO 3 against ln(C). The ε value for groundwater of each zone was estimated ( Fig. 9), which was judged as the denitrification type (Figs. 7 and 8b) under the assumption that there is no additional nitrate input at the intercept of the flow and dilution and dispersion rates were constant through the flow. We used this value to compare the relative efficiency of the denitrification activity at each flow zone. The ε value for groundwater under the Midori River watershed (shown as zone C) was also evaluated (Figs. 1 and 8 c and 9 ). As a result, a relatively higher attenuation effect by denitrification was found in the stagnant zone compared to that in the recharge zone (Figs. 2 and 9). In addition, the highest attenuation effect was confirmed in the hills-plain border zone ( Figs. 1 and 2; denitrification hotspot) even over a short distance, except around the discharge zone. This zone has the highest potential for natural attenuation of ground- water nitrate pollution through circulated groundwater systems.

6. Toward the better use of the d 15 N and d 18 O

tracer technique

Several approaches for tracing the source of groundwater ni- trate pollution have recently been proposed, for instance, the

use of chemical markers ( Fenech et al., 2012) and application of other stable isotope tracers ( Vengosh et al., 2002; Widory et al., 2004; Xue et al., 2009; Hosono et al., 2010, 2011a) in

addition to d 15 N NO 3 and d 18 O NO 3 . These tracers are definitely

useful for achieving a comprehensive understanding of the nitrate pollution mechanisms. However, in a strict sense these tracers can be useful under the hypothesis that they can be used as a proxy of nitrate. Therefore, seeking a more efficient use of nitrate isotope tracers is still important as it is the only direct fingerprint of tracing nitrate sources and recording the occurrence of denitrification.

With the traditional way of utilizing d 15 N and d 18 O tracers on d 15 N NO 3 vs. d 18 O NO 3 and NO vs. d 15 N NO 3 diagrams ( Fig. 6), it was difficult to separate the two types of anthropogenic ni- trate sources (chemical fertilizers and treated wastewater from sewage treatment plants derived through recharging river water) and also two types of attenuation processes (dilution and denitrification) for Kumamoto groundwater, since all samples are plotted within similar compositions ( Figs. 6 and 8). In contrast, the analysis taking into account detailed groundwater flows enables us to separate the effects of the mixing of different sources and denitrification ( Fig. 8). Our study clearly demonstrated that the nitrate isotopic traceability significantly improves with the understanding of groundwater flow dynamics.

3

7.

Conclusions

Our study revealed that the application of chemical fertilizers is a prime cause of groundwater nitrate pollution in Kuma- moto. Nitrogen load from manure was not detected as being the dominant source of pollution. However, this effect should be checked during future monitoring programs, since the number of livestock farms has increased in more recent years. Polluted groundwater is attenuated along the flow distance from recharge through lateral flow to the discharge (or stag- nant) zone by two different processes: dilution and denitrifi- cation. About 60 e80% and 98% of nitrate of anthropogenic origin were assumed to be purified by dilution and denitrifi- cation, respectively. Denitrification was evaluated as the more efficient attenuation function and the hills-plain border zone has the highest attenuation potential over the area. In contrast, the groundwater attenuated by dilution appears to

be more vulnerable to additional pollution. Aspects regarding

to these natural attenuation potentialities should be infor- mative for future groundwater resource exploration and preservation. This study demonstrated that traceability of nitrate isotope ratios can be improved by the combined use of land-use and groundwater flow information. The proposed approach should be applicable for future investigations in other regions.

Acknowledgments

We thank Mr. Satoshi Sakaeda and Mr. Masaaki Hirohata of the Kumamoto Prefectural Government for their help during field surveys. We also thank Prof. Yasunori Kawagoshi of the Department of Civil & Environmental Engineering, Kuma- moto University, for a fruitful discussion. Groundwater flow models used in this study were constructed under the operation of the Geosphere Environmental Technology Cor- poration, Tokyo, Japan. This study was funded by a CREST Project (JST: Japan Science and Technology Agency) with additional support from the Grant-in-Aid for Young Scien- tists (A) (no. 24681007).

Appendix 1. Figure showing increase of nitrate

concentrations (vertical axis; mg l

with time (horizontal axis; year) in confined

groundwater at selected borehole sites in

Kumamoto area (modified after Tomiie et al.,

2011 ).

L 1

as NO L N)

L

3

groundwater at selected borehole sites in Kumamoto area (modified after Tomiie et al., 2011 ). L

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