Ecological Engineering 25 (2005) 365–381

Nitrogen spiraling in subsurface-flow constructed wetlands:

Implications for treatment response
Robert H. Kadlec a,∗ , Chris C. Tanner b , Vera M. Hally c,1 , Max M. Gibbs b
a Wetland Management Services, 6995 Westbourne Drive, Chelsea, MI 48118, USA
b National Institute of Water and Atmospheric Research, P.O. Box 11-115, Hamilton, New Zealand
c Graduate Program, Biological Sciences, School of Science and Technology, University of Waikato,
Private Bag 3105, Hamilton, New Zealand

Received 30 January 2005; received in revised form 3 June 2005; accepted 5 June 2005

Abstract

Nitrogen processing in treatment wetlands was investigated by use of the stable isotope 15 N introduced as ammonium. Two
small field-scale, gravel-bed wetlands with horizontal subsurface-flow (SSF) received primary meat processing water. Four
SSF cascade mesocosms, each comprising five tanks in series, received primary meat processing water, primary dairy water,
secondary dairy water or aerated secondary dairy water. The mesocosms and one of the field-scale wetland contained well-
established bulrushes (Schoenoplectus tabernaemontani), and the other field-scale wetland remained unvegetated. The systems
were operated at steady inflows, with a nominal detention times of 4–5 days. The incoming ammonium nitrogen ranged from
18.5 to 177 g m−3 , and removals ranged from 15 to 90% for the various feed waters. Each system was dosed with a single pulse
of 15 N ammonium mixed into the feed wastewater, and the fate and transport of the isotopic nitrogen were determined. The 15 N
pulses took 120 days to clear the heavily loaded field-scale wetlands. During this period small reductions in 15 N were attributable
to nitrification/denitrification, and a larger reduction due to plant uptake. Mesocosm tests ran for 24 days, during which only
1–16% of the tracer exited with water, increasing with N loading. Very little tracer gas emission was found (∼1%). The majority
of the tracer was found in plants (6–48%) and sediments (28–37%). These results indicated a rapid absorption of ammonium
into a large sediment storage pool, of which only a small proportion was denitrified during the period of the experiment. Plant
uptake claimed a fraction of the ammonium, determined mainly by the plants requirement for growth rather than the magnitude
of the nitrogen supply. A rapid return of ammonium to the water was also found, so that movement of 15 N through the wetland
mesocosms was comprised of a spiral of uptake and release along the flow path. A two compartment model was found to
reasonably represent the isotope progress through the wetlands. First order exchanges and removals were employed in dynamic
mass balances on water and solids. It is concluded that interpretation of nitrogen dynamics in wetlands must include the nitrogen
spiral through the wetland, as well as plant uptake. This greatly increases the N residence time in treatment wetlands relative to

∗ Corresponding author. Tel.: +1 734 475 7256; fax: +1 734 475 3516.
E-mail address: rhkadlec@chartermi.net (R.H. Kadlec).
1 Present address: Air and Environmental Services Ltd., 114 Wood Bay Rd., Wood Bay, Auckland, New Zealand.

0925-8574/$ – see front matter © 2005 Elsevier B.V. All rights reserved.
doi:10.1016/j.ecoleng.2005.06.009
366 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381
the hydraulic detention time, resulting in long delays of treatment system response to changes in N loading and attenuation of
short-term fluctuations in loading.
© 2005 Elsevier B.V. All rights reserved.
Keywords: Subsurface wetlands; Nitrogen; Ammonia; Stable isotope; 15 N; Spiralling
1. Introduction
Biogeochemical cycling of nitrogen in treat-
ment wetlands is complex, involving interconversions
between different N species and transfers between stor-
age compartments (Howard-Williams, 1985; Reddy
and D’Angelo, 1994). In addition to interstitial water,
nitrogen may be stored in above and below ground, live
and dead plant material, invertebrates and microbes,
and sorbed on the surfaces of organic and inor-
ganic solids. Conversion pathways include ammo-
nia volatilization, ammonification, nitrification, den-
itrification, anammox (anaerobic ammonium oxida-
tion), dissimilatory reduction, and plant and microbial
assimilation and remineralization during decomposi-
tion (Jetten et al., 1999; Kadlec and Knight, 1996).
Sorption and microbial assimilation (including algal)
are rapid, but higher plant assimilation is slower. Plant
litter decomposition (including roots and rhizomes) is
also relatively slow, with turnover rates of months to
years. In cold climates, winter represents a period of
very limited activity, thus adding to the length of stor-
age.
Despite this complexity, most studies continue to
interpret wetland nitrogen removals on a steady-state,
‘green box’, input–output basis (USEPA, 1999, 2000).
Such analyses are confounded by temporal factors
related to the detention of nitrogen within the large
internal wetland storages, and by the confusion of sea-
sonal uptake and release vis-a-vis temperature modu-
lation of microbial processes.
It is clear that paired contemporaneous measure-
ments of input and output flows and concentrations
have little chance of providing a quantitatively accu-
rate description of nitrogen removals because of the
transport delay in the wetland. Typical hydraulic deten-
tion times are of the order of several days to more
than a week, which implies a significant shift in event
timing if the wetland is in plug flow. However, tracer
testing of hundreds of treatment wetlands has shown
conclusively that no treatment wetland exhibits plug
flow; rather, the detention time distribution extends to
three or more nominal detention times (Kadlec, 1994).
Therefore, there are remnant effects of inlet events at
the outlet after three or more nominal detention times.
Therefore, the only chance of avoiding a transport
delay artifact (synoptic error) is to compare inlet and
outlet measurements averaged over more than those
three nominal detention times. However, it is unlikely
that water detention time appropriately reflects delays
in the dynamic and stationary compartments of the
wetland—the substrate, sediments and biota. That is,
the nitrogen residence time will vary markedly depend-
ing on how many times it has been “parked” and recy-
cled in various active or passive storage compartments
during its passage through the wetland.
As a frame of reference, consider the hypotheti-
cal compartmentalization and storages for a SSF wet-
land treating secondary wastewater (Fig. 1; modified
from Tanner et al., 2002). Values were either chosen
as typical of the range of experimental observations,
or estimated (to be discussed in more detail below).
The overall nitrogen content of the various solid stor-
ages totals over 200 g N m−2 . Nitrogen removal rates
were of the order of 2 g N m−2 day−1 corresponding to
hydraulic detention times of about 5 days. Therefore,
the nominal displacement time for solid phase nitro-
gen is about 100 days. However, first order turnover
is more likely than displacement, and not all of the
solid phase nitrogen is available for turnover. Turnover
times are rapid, of the order of 1 day, for sorbed ammo-
nium (Sikora et al., 1995; Tanner et al., 1999; Triska
et al., 1994). However, plant detritus turns over much
more slowly, with half lives of up to 1 year or more
(Davis and Van der Valk, 1978; Godshalk and Wet-
zel, 1978; Hietz, 1992; Tanner, 2001). These approxi-
mations for nitrogen storage and fluxes indicate that
the nitrogen detention time in the constructed wet-
land is likely to be far greater than the water detention
time.
The importance of the large pools of solid phase
nitrogen lies in the fact that small percentage changes
 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381
Fig. 1. Nitrogen processing schematic. The numbers are storage estimates in g m−2 .
in those storages can have large consequences for
the water concentrations. For instance, the seasonal
senescence of biomass is capable of adding back a
portion of the nitrogen stored during the growing sea-
son (Sartoris et al., 2000; Tanner, 2001), thus adding
an element of seasonality to performance. There are
also large ecosystem start-up effects, due to the build-
up of biomass nitrogen, which can dominate nutrient
removal for many months (Busnardo et al., 1992; Tan-
ner et al., 1998a; Tanner, 2001).
The interactions shown in Fig. 1 occur along a spa-
tial gradient from wetland inlet to outlet. Water moves
along that gradient, and changes its chemical character
in response to the nitrogen exchanges. For instance,
a nitrogen atom that enters as ammonium may be
taken up at an upstream location, by sorption or by
biological uptake, only to be later released by des-
orption or biomass decomposition. That same atom
may later be detained again in a solid compartment,
and again released to downstream advective transport.
This stop-and-go progression or repeated biotic and
367
abiotic cycling of nutrients coupled to downstream
transport can be cast as a probabilistic time delay
model of solute transport (Buffham et al., 1970). In
the stream literature, it has been termed “spiraling”
(Elwood et al., 1983; Newbold et al., 1983), and is one
of the central concepts in the understanding of solute
dynamics (Stream Solute Workshop, 1990). Spiraling
incorporates the idea that in flowing systems, repeated
cycling of nutrients and other solutes become spatially-
displaced by downstream transport. The ‘tightness’ (or
conversely the length) of the spiral, and the intensity
and nature of the transformations that occur in each
cycle will affect the retentiveness and processing rate
of elements in the system. Howard-Williams (1985)
noted that the nutrient removal capacity of wetlands
was a function of their large storage reservoirs and short
spiral lengths.
The objective of this study was to use short-term
pulses of 15 NH4 -N stable isotopes to provide estimates
of N residence times in subsurface-flow wetlands treat-
ing wastewaters with a range of different organic matter
368 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381
and nitrogen contents. We then use the results of these
measurements to develop simple models to explore the
influence of N residence times on wetland treatment
responses.
2. Methods
2.1. Field-scale wetlands
2.1.1. Operational conditions
A 15 N tracer study was carried out in two SSF
treatment wetlands constructed by the Meat Research
Institute of New Zealand (MIRINZ) to test treatment
of meat processing wastewater. Details of wetland
design and treatment performance during the first 18
months operation have been reported by Van Oost-
rom and Cooper (1990). In brief, one wetland had
been planted with Schoenoplectus tabernaemontani,
and one left unplanted. Each was 1.5 m × 12 m × 0.8 m
deep, lined with 1.0 mm butyl rubber, and filled with
2–10 mm gravel, of 30% porosity, to a depth of 0.45 m.
The water level was maintained just below the gravel
surface.
The isotopic tracer studies reported here were per-
formed in these wetlands after 2 years operation. They
were carried out sequentially, initially in the unplanted
wetland during spring and then in the planted wetland
during late summer. During this period the wetlands
received approximately 32 mm day−1 (∼4 day nom-
inal detention time) of nitrogen-rich meat processing
wastewater; mean NH4 -N = 122, TKN = TN = 129,
TP = 17, COD = 239 g m−3 . Mean contaminant con-
centrations were reduced by passage through the
wetland to: NH4 -N = 104, TKN = TN = 105, TP = 14,
COD = 73 g m−3 , and NH4 -N = 102, TKN = 103,
TN = 105, TP = 13, COD = 72 g m−3 for unplanted and
planted wetlands, respectively.
2.1.2. Experimental technique
Isotopically enriched water was fed to each wet-
land, as 150 g m−3 of NH4 -N enriched to 10% excess
15 N. An 8-h pulse was used, at the normal hydraulic
loading rate. The channels were tested over periods
of 110 (unplanted) and 133 (planted) days, the time
required to completely pass measurable tracer pulses.
Mass spectrometry was used to determine 15 N. Sam-
ples were analyzed for 15 N using a train consisting of a
Carlo Erba NA1500 CNS elemental analyzer, followed
by a gas chromatograph, then by a Europa Tracermass
Stable Isotope Analyzer (Hally, 1990). The CNS ana-
lyzer combusts the sample, and reduces all nitrogenous
gas to dinitrogen (N2 ), the chromatograph isolates all
nitrogen isotopes (molecular masses 28, 29, 30), and
the mass spectrometer provides the fractions of these
various stable isotopes.
Water samples were taken by syringe from depths
of 20 and 40 cm, at 1 m intervals along the wetlands
on 9 (unplanted) and 13 (planted) dates during pulse
passage. Water samples were dried and the residues
analyzed for 15 N as solids.
Dissolved gases were sampled at depths of 5, 30, and
40 cm at distances of 2, 7, 10, and 12 m, on 13 dates
(planted and unplanted) during pulse passage. Acid-
ified samples were analyzed for 15 N by a variant of
Hall’s (1980) headspace technique. Gases in solution
were equilibrated with an argon atmosphere, which was
analyzed for the sum of N2 and N2 0, via copper reduc-
tion followed by gas chromatography. 15 N was then
determined by mass spectrometry.
Organic sediment samples were taken, after the
water-borne pulse of 15 N had entirely passed, from
three depths in each wetland, in the root zone, below
the root zone and at the wetland bottom. Samples
of gravel and solids were taken at longitudinal dis-
tances of 2.5, 6, and 10 m (unplanted) and 2.5, 7, and
12 m (planted). The sediments were rinsed from the
gravel, filtered and dried prior to the analysis described
above.
Plant roots/rhizomes, old culms and new culms were
harvested at 1 m intervals along the wetland at nine
times during the course of pulse travel. These were
processed and analyzed according to the procedures
for solid samples given above.
2.1.3. Ancillary testing
Separate, exploratory batch tests were conducted
to assess nitrification and denitrification routes, and
plant uptake. Nitrification and denitrification within the
gravel substrate was studied in jar tests. These involved
adding 30% excess 15 N of either (NH4 )2 SO4 or NaNO3
to 0.35–0.40 L of removed water and gravel. Atmo-
spheres of pure argon and 21% oxygen in argon were
imposed in separate replicated experiments. Headspace
samples were analyzed for 15 N in the sum of dinitrogen
and nitrous oxide.
 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381
Plant uptake was investigated in bucket tests, into
which about 12–14 L of plants, gravel and water were
transferred. The buckets were bottom-fed with effluent
at regular intervals. The test was conducted by introduc-
ing 20 g of (NH4 )2 SO4 10% excess 15 N in 4 L (one pore
volume) of water. After 24 h, the bucket was flushed
with four pore volumes of water (15 L). Evolved NH4 -
N gas was chemically trapped with HCl, and analyzed
for 15 N. Plants were divided into new and old culms,
and roots plus rhizomes, and dried and analyzed for
15 N. Sediments were washed from the gravel, dried
and analyzed for 15 N.
2.2. Cascade mesocosms
2.2.1. Operational conditions
The mesocosm wetlands, their operating conditions
and treatment performance have been described in
detail in Tanner et al. (2002). Here we provide a
brief synopsis of those conditions, as they form the
background against which the 15 N experiments were
conducted.
Eight parallel wetland cascade mesocosms were
constructed, with adjacent pairs receiving the either
primary treated meat processing wastewater (M1), pri-
mary treated farm dairy wastewater (D1), secondary
treated farm dairy wastewater (D2) or aerated sec-
ondary treated farm dairy wastewater (D2A). Each
consisted of a series of five inter-connected, 20 L white
PVC tanks arranged down a series of tiered platforms.
Each tank was filled with 15 mm greywacke gravel to
within 20 mm of the top (8 L initial void volume) and
planted with five propagules of the native soft-stemmed
bulrush, Schoenoplectus tabernaemontani, in January,
1996. The isotope tests were conducted in November
1996, at which time mature plants were present in the
tanks.
Each tank had a 32 mm i.d. PVC pipe inserted at the
side which extended to the base, prior to filling with
gravel, to allow addition of wastewater. Wastewater
thus entered at the base, creating an upflow movement
of water through the root zone of the plants. An outflow
tube was placed in the side of the tanks, immediately
below the gravel surface, opposite to the inflow tube,
and displaced wastewater flowed into the base of the
subsequent tank.
Four times daily (4 a.m., 10 a.m., 4 p.m. and 10
p.m.) wastewater was pumped into delivery vessels
369
suspended above the upper tank of each cascade meso-
cosm. Wastewater remained in the delivery vessels for
approximately 2 min after pumping had ceased, before
electronic valves at the base of each bottle opened,
allowing wastewater to enter the first tank in each
cascade. The wastewater entered at the base of each
mesocosm, displacing wastewater from the top, which
flowed into the inflow tube of the next tank, and so on
down the cascade.
The cascades were supplied with 8 L day−1 wastew-
ater from January until June, when flow was altered to
2 L day−1 wastewater until August for the winter low
growth period of the bulrushes. Flow was then returned
to 8 L d−1 wastewater. In late October, 2 weeks prior
to the start of the sampling period, 1000 L of each
wastewater was collected and stored in sealed con-
tainers under refrigerated conditions (4–5 ◦ C) so that
a single batch of wastewater could be used through-
out the sampling period. Wastewater was transferred
to the refrigerated container adjacent to the cascades
as required. At this stage, a 80 mL plastic container
was inserted into the surface of the gravel to the
depth of the wastewater in each mesocosm adjacent
to the outflow point, to act as a sample collection
vessel.
A clear, gas-impermeable plastic sleeve (Trygon
Vapour Barrier) was sealed to the rim of each tank
with a large rubber band, to form a gas sampling “bag”
type enclosure. When required for gas sampling, the
sleeving was pulled up over a 0.5 m high frame of
galvanised iron rod, and closed by folding the end
of the tube over several times and sealing with par-
cel tape. The enclosure was left overnight (to reduce
radiative heating effects), and a gas subsample removed
the next morning. When not in use (during the day-
time), the plastic sleeve was folded down like a stocking
around the top of the tanks leaving the plants fully
uncovered. The top of the frame was circular (300 mm
diameter), the same as the top of the tanks giving a
volume of the enclosure of 0.035 m3 (including plant
stems).
Intensive sampling was carried out early in
the Southern Hemisphere summer (12 November–6
December 1996). On the first day of sampling, one of
each pair of cascades was also dosed in their 10 a.m.
wastewater addition with 0.485 g of 15 N as NH4 Cl, and
0.1 g of Br as KBr. The KBr spike was used as an inert
flow tracer, and the 15 N labelled NH4 Cl as a marker for
370 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381

nitrogen transformations within each mesocosm of the 2.2.4. Plant samples

cascade. On each sampling day, including Day 0, one actively
growing stem of bulrush was cut at the gravel surface
from each tank of the four cascades which had been
2.2.2. Gas sampling
dosed for 15 N. The bases were rinsed in clean water
On the evening prior to each sampling day (at 4
and the lower 5 cm of that stem placed in a glass screw
p.m.), except Day 0, the plastic enclosures were raised
capped vial and frozen pending analysis of 15 N, %N
and sealed over the plants and left sealed overnight.
and %C.
On the morning of each sampling day at approxi-
As the reeds continued to grow throughout the
mately 9 a.m., a 60 mL air sample was drawn into
experiment and would have exceeded the height of the
a syringe, via a needle inserted through the wall of
gas sampling enclosures, the tops of the leaf stems
the enclosure. The syringe was flushed twice, and
were clipped at 3–4 day intervals during the sam-
then withdrawn. The needle was then pushed into a
pling period back to the starting height of 40 cm set
15 mL draw vacutainer (Becton Dickson brand, red
on Day −1. Trimmings were collected, dried in a
top) and allowed to suck in the sample to approx-
forced-air drying oven at 85 ◦ C to constant weight and
imately ambient pressure. A second syringe needle
weighed.
was inserted through the vacutainer septum and the
syringe emptied into the vacutainer allowing excess
2.2.5. Final destructive sampling
gas to vent from the second needle. Both needles were
At the end of the trial, all the plant stems were
withdrawn together. The procedure was then repeated
removed from each mesocosm, counted, and wet
for a second vacutainer and samples stored for later
weighed. A 30 stem subsample was wet weighed, and
analysis for 15 N2 and 15 N2 O. The plastic enclosures
then its volume determined by displacement of water
were then opened and lowered, until the next sampling
in a 500 mL measuring cylinder. Subsamples were then
event.
dried in a forced-air drying oven at 85 ◦ C, prior to re-
Samples were taken for 15 N analysis on days 1–5,
weighing.
7, 9, 11, 14, 17, 21, and 25 after the initial dosing (13
The void volume of the mesocosm was then deter-
November–6 December 1996, Southern Hemisphere
mined prior to sub-surface sampling by draining and
summer). Analysis of 15 N and 14 N stable isotopes was
measuring the volume of interstitial water. Each meso-
performed using a continuous flow isotope ratio mass
cosm was then emptied into a large flat container
spectrometer (Finnigan MAT Delta-plus, Bremen, Ger-
and divided vertically with a serrated-edged knife and
many) calibrated against ultra-pure grade standards
biofilm samples removed (see below). The entire meso-
(0.36625 at.% 15 N). The total N content of samples
cosm was then sorted for plant matter by removing
was determined relative to an acetanilide standard with
sub-surface plant stems, rhizomes and roots. Detached
10.36% N.
roots were floated out with water and removed with
a sieve. The entire subsurface plant sample was then
2.2.3. Water sampling placed in a forced-air drying oven at 85 ◦ C until dry,
Water samples were collected from the 80 mL plas- and then weighed.
tic container set into the gravel in front of the outlet tube
on days 1–4, 6, 8, 10, 13, 16, 21, and 25 after initial dos- 2.2.6. Biofilm sampling
ing. The sampling container was emptied of residual At the time of destructive sampling of the meso-
water immediately prior to the wastewater addition at cosms, gravel samples were taken from the top, mid-
10 a.m., allowing fresh effluent from the mesocosm to dle, and bottom (approximately, 20–70, 120–170,
be sampled just after the cascade cycled. A 10 mL sam- and 220–270 mm depths) of the mesocosm. Biofilm
ple was placed in a 30 mL scintillation vial and stored at cover estimates were made by observing the gravel
4 ◦ C pending later 15 N analysis as outlined above. Con- under a binocular microscope. Total cover was esti-
tainers (2 L) placed at the end of the cascade were used mated, and then biofilm thickness was measured under
to measure actual outflow volumes during the sampling higher magnification against a calibrated eyepiece
period. graticule.
 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381 371

3. Results

3.1. Field-scale wetlands

3.1.1. Plant uptake, gaseous emission, and

sediment detention assays
The batch jar tests showed that added tracer nitrate
was denitrified, as determined from the detection of
15 N in the headspace of either aerobic or anaerobic

atmospheres, in increasing amounts over the 120 h

duration of the test. Conversely, the addition of tracer-
tagged ammonium resulted in virtually no 15 N in the
headspace of either aerobic or anaerobic atmospheres. Fig. 2. Response of planted and unplanted wetlands to an impulse
Thus the indications were that nitrification was rate- of 15 NH4 .
limited in both aerobic and anaerobic environments
(Hally, 1990). Table 1
Mass balance of ammonia 15 N in pulse tests of planted and unplanted
The bucket tests showed that added ammonium 15 N SSF wetland channels
was found in all plant parts after 24 h of exposure,
Planted Unplanted
and in about equal proportions (30 mg to old culms,
42 mg to new culms, and 46 mg to roots and rhizomes). g % g %
The total rate of nitrogen uptake, for isotopes 14 and Dose 5.000 5.000
15, was 1.7 g m−2 day−1 for this test. Sediments were Flow out 3.518 70 5.055 101
found to contain a similar amount of 15 N, 106 mg after Volatilization 0.002 0 0.002 0
24 h. Essentially no volatilized ammonia was mea- Sediment storage 0.073 1 0.027 1
sured, nor any labelled gaseous denitrification prod- Denitrification 0.083 2 0.085 2
Plant storage 0.966 19 0.000 0
ucts. The implication is that plant uptake of ammonium
is an active removal mechanism, and that all plant parts Total out 4.642 93 5.169 103
are reservoirs for nitrogen storage. Similarly, ammo-
nium 15 N was trapped in sediments.
Table 2
Biomass, nitrogen contents, and stocks of 15 N (mg m−2 ) at final
3.1.2. Tracer passage through the wetland
harvest
The 15 N detention time of the wetland channels was
Distance (m) New shoots Old shoots Root/rhizome Total
surprisingly long compared to water detention time
(Fig. 2). The unplanted channel had a mean 15 N deten- Biomass (g m−2 )
2.5 20 518 790 1328
tion time of 51 days and the planted channel 64 days,
7 9 671 636 1316
compared to the water detention time of ∼4 days. 12 19 661 1343 2023
Recovery of 15 N in the outflow water was 101% for
N content (%)
the unplanted channel, and 70% for the planted chan- 2.5 5.7 2.6 3.2
nel (Table 1). Based on the assay results very little 7 4.7 1.9 2.1
of the labeled NH4 -N was lost via volatilization, or 12 3.9 1.7 2.2
nitrification/denitrification. Only small amounts of the 15 N content (at.% excess)
tracer remained in sediments after the tracer pulse had 2.5 0.0359 0.0258 0.0205
passed, and that was concentrated near the inlet end of 7 0.0269 0.0159 0.0147
the channels, as determined from final destructive sam- 12 0.0092 0.0043 0.0058
pling. About 20% of the 15 N was found in various plant 15 N storage (mg m−2 )

parts at final testing (Table 2). Because the final harvest 2.5 0.422 3.960 5.193 9.574
was in a period of plant senescence, most of the above- 7.0 0.107 1.941 2.205 4.253
12.0 0.067 0.452 1.713 2.233
ground biomass was in the form of old culms. Most of
372 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381

the 15 N was found in those old culms and in roots and

rhizomes.
The short duration pulse of 15 N created the bell-
shaped response curves in Fig. 2. The inlet dose may be
regarded as an impulse, lasting 0.33 days in comparison
to the approximately 100 day response. Therefore, the
curves may be regarded as the detention time distribu-
tion for nitrogen in the wetlands. A gamma distribution
fit of each response is shown, of the form:

N−1

1 t t
g(t) = exp − (1)
ti Γ (N) ti ti
where t = detention time (d), ti = τ/N, τ = mean deten-
tion time (d), N = shape parameter, Γ (N) = gamma
function of N,
 ∞
Γ (N) = xN−1 e−x dx (2)
0

When N is an integer, the gamma function is

Γ (N) = (N − 1)! (a factorial). When N = 1, the gamma
distribution becomes the exponential distribution. Both
the gamma distribution and the gamma function are
readily available as computer spreadsheet tools (e.g.,
GAMMADIST and GAMMALN in ExcelTM ). Eq. (1)
may be easily fit to tracer data by selecting N and ti
to minimize error (e.g., SOLVER in ExcelTM ). This
is gradient search procedure, in which N and ti are
selected to minimize the sum of the squared errors
(SSQE) between the tanks-in-series (TIS) model and
the data.
The best fit parameters for the measured distribu-
tions of nitrogen detention times are:
• Unplanted: τ = 51 day, N = 5.5, R2 = 0.88
• Planted: τ = 64 day, N = 6.4, R2 = 0.75
These must be contrasted to the nominal water
detention time of about 4 days. Any water that did not
interact with wetland sediments or biota should have
exited the wetland prior to the first sampling on day
10. However, that water response may not have been
entirely flushed in the case of the unplanted wetland,
which shows anomalously high 15 N on days 10 and
12. However, closure of the 15 N mass balances indi-
cates that nearly all the 15 N interacted with sediments
Fig. 3. Progress of the internal peak in water 15 N concentration along
the field-scale wetland channels.
inlet to the outlet (Fig. 3), decreasing in magnitude as it
traveled. Water column measurements indicate that 15 N
denitrification products and 15 N NH4 -N passed through
the inlet section of the wetland (Fig. 4). Such peaks
traveled down the wetland, and became more dispersed
as time progressed. There was more denitrification in
the unplanted wetland, as evidenced by the larger peak
in Fig. 4.
A distinct gradient of decreasing plant storage was
found from inlet to outlet (Table 2). Plants in inlet
zone had the first opportunity to access15 N, and at the
highest concentrations. The disproportionate amount
of 15 N in the old (senesced) plant parts is due to the
time of harvest, which was well after the peak of the
growing season. Gradients in biomass, total nitrogen
content and 15 N are compounded into the areal stor-
age, with total nitrogen and 15 N decreasing from inlet to
outlet.
End-of-test sediment sampling indicated very little
15 N in that form of storage (Table 1). The unplanted

and plants (Table 1). system showed an increasing trend in 15 N from inlet to
The internal location of the 15 N ammonium peak outlet, and more near the bottom (Table 3). In contrast,
moved gradually in the water compartment, from the the planted system had little or no longitudinal gradient,
 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381 373

internal gradients may be found in Tanner et al. (2002).

In general terms, the primary influents M1 and D1 were
very strong, and experienced significant reductions of
ammoniacal nitrogen and COD along the cascades.
The inlet zones of these mesocosms had accumulated
large quantities of sediment and biofilms, occupying
20–30% of the gravel void volume. Mesocosms D2
and D2A received secondary and partially nitrified sec-
ondary effluents, and accumulated lesser amounts of
sediments and biofilms. However, the mesocosms con-
tained as much or more sediments and biofilm solids
than the field-scale wetlands.
The same may be said for plant materials. The end
of experiment harvest of the mesocosms showed stand-
ing crops of 465 g m−2 above ground and 3035 g m−2
below ground, compared to field-scale standing crops
of 635 g m−2 above ground and 925 g m−2 below
ground.
Accordingly, the inventory of exchangeable or
cycleable nitrogen in the mesocosm solid compart-
ments was somewhat greater than that in the field-
scale wetland on a unit area basis. The nominal deten-
Fig. 4. Time progression of dissolved 15 N species at 2 m (15 NH4 - tion times were close; 4.5 day for the mesocosms and
N) and 3 m (gaseous 15 N denitrification products). The water travel 4.0 day for the field-scale wetlands. Based on the results
times are approximately 16 and 24 h, respectively.
for the field-scale wetland, the 15 N detention time (rel-
but considerably more 15 N in the root zone than below ative to the hydraulic detention) would be expected to
it. be around 40–50 days. It is, therefore, likely that the
mesocosm 15 N pulses would have been only partially
3.2. Mesocosm wetlands expelled over the 24 days of monitoring.
This approximation was borne out by laboratory
The general performance characteristics of the four results for 15 N in the effluent waters from the meso-
cascade mesocosms are given in Table 4. Details of cosm tanks (Fig. 5). For water samples, the back-
ground determined from both standards and blanks
Table 3 was 0.36625 at.% 15 N. Excess 15 N was determined by
Sediment content of 15 N (at.% excess) at final harvest
difference, and attributed to the isotope addition. For
Distance (m) D2 and D2A, which had lower strength influents and
2.5 7 12 Mean lower amounts of sediments and biofilms, 15 N peaks are
Planted observable for the first two or three tanks. However, 15 N
Root zone 0.0100 0.0205 0.0184 0.0163 was essentially undetectable in the waters in the outlet
Below roots 0.0092 0.0120 0.0099 0.0104 regions. The D1 mesocosm shows a much accelerated
Bottom 0.0050 0.0055 0.0051 0.0052 peak passage, presumably due to the ca. 40% void
Average 0.0081 0.0127 0.0111 blockage by solids, coupled with ca. 25% void blockage
2.5 6 10 Mean by roots. M1 shows erratic behavior, for which there is
no apparent explanation. It should be borne in mind that
Unplanted
the water analysis does not measure dissolved gaseous
Top 0.0024 0.0109 0.0205 0.0113
Bottom 0.0052 0.0180 0.0228 0.0153 nitrogen (N2 ) or nitrous oxide (N2 O). Therefore, some
isotope could have exited, as dissolved products of
Average 0.0038 0.0145 0.0217
denitrification, undetected by the analytical techniques
374 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381

Table 4
Mesocosm performance during 15 N testing (Tanner et al., 2002)
Concentration (g m−3 ) M1 D1 D2 D2A

In Out In Out In Out In Out

COD 383 134 2240 658 438 332 272 132
TKN 199.1 126.7 135.2 33.7 40.4 11.1 32.4 5.0
NH4 -N 176.7 114.6 118.7 13.8 23.3 2.1 18.5 1.9
TN 199.8 137.4 137.5 41.2 42.2 12.2 54.4 5.7
NOx -N 0.7 10.7 2.3 7.5 1.8 1.1 22.0 0.7

Mass flux (g m−2 day−1 ) Loading Removal Loading Removal Loading Removal Loading Removal
COD 8.7 6.50 47.3 35.54 9.7 5.09 5.8 3.84
TN 18.5 2.28 11.9 2.38 2.3 0.79 3.2 1.15

employed here. The time-integrated amounts of 15 N
measured in the exit waters were 15.5 and 11.8% for M1
and D1, respectively. Lesser fractions departed from D2
and D2A, 2.1 and 0.7%, respectively (Table 5).
The gaseous 15 N background was taken as the mean
for 4 days in the un-dosed mesocosms, which had a
global average of 0.36818 at.% 15 N, similar to the nor-
mal abundance in air (0.3663 at.%). Excess 15 N was
determined by difference, and attributed to the iso-
tope addition. Gas samples showed no particular time
trend, nor any gradient along the cascades, nor any dif-
ferences between mesocosms (different influents) for
excess isotope. Overall very little of the added iso-
tope was detected in gaseous emissions from any of
the mesocosm tanks (Table 5).
Indications are, therefore, that the large majority of
the isotope was trapped in the mesocosms, and had
not exited by the end of the sampling period, either as
gases or dissolved nitrogen salts. Consequently, much
of the added tracer remained in the wetland meso-
cosms, associated with plants and sediments. Plants
were separated into above and below ground parts, and
analyzed for 15 N. The above ground 15 N background
was taken as the average value for each wetland at the
start of the experiment; M1 = 0.38733, D1 = 0.37166,
D2 = 0.42018, D2A = 0.37461 at.% 15 N. These differ-
ent background levels of isotopic N enrichment in the
plant tissues are likely to relate to differences in iso-
topic abundance in the different wastewater sources,
as well as microbial and physico-chemical discrimina-
tion against the heavier 15 N isotope during preceding
treatment stages, and (to a lesser extent) within the

Table 5
Fate of added 15 N in mesocosm trains, as percent of added isotope
M1 D1 D2 D2A
Roots 5.6 11.9 36.4 45.6
Above ground 0.2 0.7 2.1 1.6
Sediments 28.2 34.8 31.5 36.4
Total solids 34.1 47.4 70.1 83.6
Exit water 15.5 11.8 2.1 0.7
Remaining water 1.1 2.1 0.5 0.4
Total water 16.5 13.9 2.6 1.1
Solids and water 50.6 61.3 72.7 93.4
N2 gas out 0.6 0.4 2.6 0.8
Measured out 51.3 61.8 75.3 85.5
Fig. 5. Longitudinal gradients in the storage of added 15 N in above
(solids, gas, water)
ground plant tissues along the mesocosms.
 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381
wetlands themselves (Knowles and Blackburn, 1993;
Robinson, 2001). Excess 15 N was determined by dif-
ference, and attributed to the isotope addition. Despite
the fact that only small amounts of the added 15 N were
found in above ground plant tissues (Table 5), some
interesting features emerge. Firstly, a lower percentage
of added isotope was found in above ground plant mate-
rial in the heavily loaded mesocosms (M1 = 0.2% and
D1 = 0.7%) than in the lighter loaded ones (D2 = 2.1%
and D2A = 1.6%). This is due both to the greater dilu-
tion of added 15 N within the larger sediment N pools
that had accumulated in the highly loaded M1 and
D1 wetland mesocosms (Tanner et al., 2002), and the
limitations of plant uptake, related to growth require-
ments within the available physical space and the flux
per unit area of incident sunlight. Plant uptake, there-
fore, exerts less influence in systems with high nitro-
gen supply than it can on systems with low nitrogen
supply. Secondly, there are pronounced gradients in
above ground storage, progressing downward in the
flow direction (Fig. 5). There were no significant gra-
dients in biomass, nor differences in biomass among
mesocosms (P < 0.05). Therefore, the isotope gradients
are primarily the result of different tissue concentra-
tions of 15 N, showing that within the period monitored,
most of the 15 N taken up by plants had been ‘captured’
in the first few tanks of the mesocosms.
Analysis of below ground plant material, roots and
rhizomes, is complicated by difficulties in separating
out media, sediments and biofilms. Additionally, no ini-
tial measurements of background concentrations of 15 N
could be taken without tank destruction. Background
concentrations were, therefore, taken to be the lowest
found, in the final destructive sampling of the respec-
tive trains: M1 = 0.39830, D1 = 0.38061, D2 = 0.42489,
D2A = 0.40614 at.% 15 N. Excess 15 N was determined
by difference, and attributed to the isotope addition. A
lower percentage of added isotope was found in above
ground plant material in the heavily loaded meso-
cosms (M1 = 5.6% and D1 = 11.9%) than in the lighter
loaded mesocosms (D2 = 36.4% and D2A = 45.6%).
As for above ground tissues, a greater percentage of
added isotope was found in the mesocosms treating
secondary effluents. Spatial variability along the meso-
cosms was generally too high to ascertain longitudinal
trends.
Sediments were also prone to great difficulties in
homogeneous and representative sample acquisition.
375
Again, no initial measurements of background concen-
trations of 15 N could be taken without tank destruction.
Background concentration was, therefore, taken to be
the lowest found in any tank in any train, 0.38200 at.%,
at the final destructive sampling. The amount of sedi-
ment collected was typically higher in the inlet tanks
for the M1 and D1 mesocosms, but not for the D2 and
D2A. The volumes of material collected, expressed as a
fraction of the nominal pore volume, were: M1 = 20%,
D1 = 27%, D2 = 6%, and D2A = 14%. The amount of
the added 15 N found in the sediment at final sampling
ranged from 28.2 to 36.4% (Table 5). The total recov-
eries of isotope were less than 100%, and increased
progressively from M1 to D1 to D2 to D2A. Given
the uncertainties in sampling and analytical procedures,
these recoveries were deemed acceptable.
3.3. Mass balances and tracer speeds: field-scale
A simplified concept of nitrogen tracer move-
ments in the wetland environment is possible when
some of the more minor inter-compartmental trans-
fers are ignored, and the solids compartments are
lumped (Fig. 6). This simplified model allows for uni-
directional plant uptake, reversible exchange with wet-
land solids, and microbial conversion of ammonium
to nitrogen gases. It has been calibrated to treatment
of dairy wastewater with high ammonium in gravel-
bed mesocosms with good success (R2 = 0.97–0.99)
(Tanner et al., 1999). However, that calibration was
for batch operation, whereas here we are interested
in the dynamics of tracer movement through a spa-
tially variable, longitudinal SSF wetland. The spatial
and dynamic equations are:
∂C ∂C
ε +u = −ku C + kr X − kg C (3)
∂t ∂y
∂X
(1 − ε) = ku C − kr X (4)
∂t
where C = water phase concentration (g m−3 ),
kg = growth and conversion rate constant (d−1 ),
kr = release rate constant (d−1 ), ku = uptake rate con-
stant (d−1 ), t = time (d), u = water velocity (m day−1 ),
X = solid phase concentration (g m−3 ), y = distance
(m), ε = porosity.
It should be noted that a volumetric solid phase
concentration (X) has been used. It is related to the
376 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381

Fig. 6. Simplified short-term nitrogen processing schematic for high ammonium loading and low removal.

commonly-used mass concentration by: Table 6). The remaining two are determined from the
isotopic tracer experiment.
X = ρ · CS (5) There are several results of interest from this simpli-
fied model. It may be used to examine the progress of a
where CS = solid phase concentration (g kg−1 ),
tracer through the ecosystem, although nitrogen is not
ρ = solid density (kg m−3 ).
conservative. Nitrogen may be moved into solid com-
The solid phase consists of media (rock), sedi-
partments and later released back to the water, resulting
ments, organic detritus and biofilms, each of which
in a prolonged detention in the wetland. Theoretically,
has an associated exchangeable nitrogen content. These
some nitrogen may proceed directly to the outlet, with a
have been aggregated, with a mean density and a
transit time corresponding to the water detention time,
mean concentration. Measurement of this pool of
but if uptake is rapid, that water transit time peak will
nitrogen is difficult, and therefore, the solid density
be immeasurable. Therefore, the response of the wet-
is regarded as a bounded but adjustable parameter.
land to an impulse of 15 N tracer injection is expected
The net consumption of nitrogen by plant growth has
to be a bell-shaped curve, characterized by the nitrogen
been aggregated with net microbial transformations.
detention time in the solids compartment.
In a short-term tracer experiment, any tracer taken up
Fig. 7 represents a rough calibration to the field-
into vegetation will not likely be released, thus per-
scale wetlands. About 85% of the incoming tagged
mitting the assumption of an irreversible removal to
ammonia is not converted, and ultimately exits the
growth. For longer model applications, in which vege-
tation dieback occurs, a third equation similar to (4)
must be added to account for the dynamics of the Table 6
Field-scale wetland performance during 15 N testing (Van Oostrom
vegetation storage, and a vegetation N-release added and Cooper, 1990)
to (3).
Influent Schoenoplectus Unplanted
The model was implemented in a five compartment
finite difference setting, which acknowledges the like- COD (g m−3 ) 239 72 73
TKN (g m−3 ) 129 103 105
lihood that the field-scale wetlands could be hydrauli- NH4-N (g m−3 ) 122 102 104
cally represented by five tanks in series. There are three TN (g m−3 ) 129 105 105
adjustable parameters: ku , kr , and kg . One of those (kg ) COD mass loading/ 7.6 5.3 5.0
is determined by the global rate of removal of ammo- removal (g m−2 day−1 )
nium, which is determined from the previous steady TN mass loading/ 4.1 0.8 0.5
removal (g m−2 day−1 )
operating data for the wetlands (about 12–20%, see
 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381
Fig. 7. Relative tracer concentration in water and sediment stor-
age calculated from an uptake, release and removal model. The 15 N
detention time is approximately 50 days.
system. We also note that only a tiny portion of the
tagged ammonium nitrogen goes directly to the wet-
land exit (<1%). The 15N detention time for the field-
scale wetlands is about 50 days, or 12 times the water
detention time. Although the data are not sufficiently
accurate to allow precise parameter estimation, approx-
imate values were determined. The rate of ammonium
uptake was quite rapid, to account for observed tracer
response, with ku ≈ 1.0 day−1 . However, almost all
of that ammonium, ca. 99%, is eventually returned
to the water column, with the small difference com-
prising the net removal of ammonium by plants and
nitrification–denitrification. Turnover of the solid nitro-
gen storage is slow, at kr + kg = 0.17 day−1 .
Both the mean and maximum values for solid and
water phase tagged nitrogen storages decline along the
flow path. The result is that plant uptake would also fol-
low that trend, with high uptake near the inlet end. Thus
it would be anticipated that the transfer to vegetation
(irreversible within the timeframe of the experiment)
would result in end-of-test accumulations near the inlet
end of the wetland.
The long delay of the tracer, compared to the water
transit time, results from its interaction with a very
large storage pool of actively interchanging, solid phase
nitrogen. In the process, it is subject to a correspond-
ingly high dilution by that internal nitrogen pool. As a
result, high tracer pulse concentrations are drastically
reduced, by more than a factor of 100 for both model
and field results, compared to inlet concentrations. The
size of the exchangeable ammonium nitrogen pool may
377
be estimated from model Eq. (4), for the stable equi-
librium exchange condition. The partition coefficient
is given by:
Xe ku
K= = (6)
Ce kr
where the subscript “e” refers to a local equilib-
rium between water phase and solid phase exchange-
able ammonium. The response of Fig. 4 corre-
sponds to K = 6. For a water phase ammonium of
100 g N m−3 , the active solid phase ammonium would
be 600 g N m−3 . From (5), and a solid density of
2500 kg m−3 , there should be 240 mg N kg−1 of solids.
This pool of ammonium may be located in any of the
three principal components of the solid wetland matrix:
sorbed to the gravel media and organic matter, in
labile biofilms, and in high-turnover organic sediments.
Ammonium sorption studies on similar rock media
(Sikora et al., 1995; Tanner et al., 1999) suggest that
sorbed ammonium is not likely to account for the entire
storage, because only 20–30 mg N kg−1 were sorbed at
water phase ammonium of 100 g N m−3 in those stud-
ies. However, Tanner and Sukias (1995) found large
amounts of nitrogen associated with organic matter
in the void spaces of a SSF wetland treating high
ammonium wastewaters, which after 5 years operation
(C.C. Tanner, unpublished data) ranged from 100 to
500 mg N kg−1 of gravel media, representing an aver-
age accumulation of ∼200–300 g N m−2 of wetland.
This material would be attributable to a combination
of biofilms, organic detritus and sediments, at a dry
weight concentration of 2–4% nitrogen.
These quantities may be translated to an areal basis,
given the wetted depth of 0.4 m, porosity of 35% and
bulk density of 2500 kg m−3 for the field-scale system
in the present study. The projected active nitrogen pool
is 156 g N m−2 . At 4% N, this represents 3.9 kg (dry
weight) organic matter per m2 of media, which is simi-
lar to the mean organic matter accumulations recorded
after 2 years operation in comparable systems (Tanner
and Sukias, 1995; Tanner et al., 1998b).
4. Discussion
Dissolved dinitrogen and nitrous oxide gases in the
water moving through the wetland may represent an
important transport mechanism, because of their high
378 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381
solubility. Nitrogen is about twice as soluble as oxygen
and nitrous oxide about 80 times. However, the amount
of nitrous oxide is expected to be far less than that of
nitrogen, due to the atmospheric dominance of N2 . The
assumption that all nitrogen removal results ultimately
from denitrification, with production of N2 , implies
production rates in the range of 0.5–2.5 g m−2 day−1
(Tables 4 and 5). The air N-saturation content of a
0.4 m water column at 35% porosity is approximately
2.0 g m−2 . Consequently, the turnover time for dis-
solved dinitrogen would be on the order of 1 or 2
day. Given a 4–5 day water detention time, most of the
production of 15 N2 would be as gas emission. Nitrous
oxide differs, in the fact that there is presumably little
or no N2 O-N in the incoming water, or in the air with
which the water is in contact. Therefore, nitrous oxide
will accumulate in the water in which it is produced
until the gaseous loss to the atmosphere balances the
rate at which it is produced. Mass transfer to or from
water in SSF systems is relatively inefficient, and there-
fore, loss to the atmosphere is expected to be small.
For an estimated overall mass transfer coefficient of
0.02 m day−1 , estimated from free water surface reaer-
ation (Kadlec and Knight, 1996), two-thirds of the
nitrous oxide would leave in the dissolved form. This
form of (unmeasured) export may contribute to the lack
of closure of the 15 N mass balances (Table 5).
It is necessary to keep in mind the fact that the field-
scale wetlands received very high loadings of nitrogen
(predominantly in organic and ammoniacal forms) and
organic matter. Consequently N removals were low on
a percentage basis, but not on an absolute basis. There-
fore, unlike SSF wetlands loaded at lower rates more
common for secondary sewage wastewaters, 15 N losses
to microbial and plant growth processes did not have
a large effect on the bulk of the 15 N passing through
the wetland. The long delay of the isotope in the wet-
land indicates that ammonium is being processed many
times over during transit, due to microbial and sorption
processes. Conversely, the uptake by Schoenoplectus
was not reversed during the period of the isotope exper-
iment, and plant storage represented a net loss from the
water during that time frame. Presumably, decomposi-
tion would eventually return some fraction of the 15 N
in plant tissues.
The mesocosm results further highlight the dual
influence of plant uptake and sediment exchange.
Biomass was approximately constant across the four
systems, but the removal of 15 N to plant tissues was
about five times higher for the two lighter loadings
(D2 and D2A) than for the heavier loadings (M1 and
D1). The 15 N dose was the same for all, but was dis-
persed in about five times as much total nitrogen in the
cases of the heavily loaded mesocosms. If the plants
drew equally on both nitrogen isotopes, the observed
isoptope uptakes were consistent with the proportion-
ate availability.
The fraction of the applied 15 N that was found
associated with mesocosm sediments was relatively
independent of the type of feed water (Table 5). The vol-
umetric amounts of sediments recovered at end-of-test
sampling were not greatly different among mesocosms.
Although these sediments were not dried and weighed,
the amounts of solids were qualitatively comparable
to those found in studies of similar wetlands treating
dairy wastewaters (Tanner and Sukias, 1995; Tanner
et al., 1998b), which were of the order of 3000 g m−2 .
The nitrogen content of these sediments is moderately
high, ranging from 1–3% dry weight for the field-scale
wetlands. The intensity of N spiraling may be greater in
wetlands, such as those tested in the present study, that
received agricultural wastewaters with relatively high
organic matter and solids loadings. Large consequent
accumulations of organic matter within the media of
such SSF wetlands are likely to increase their cation
exchange capacity.
Turnover times of ammonium in wetland sediments
has been the subject of prior research. Bowden (1984)
studied ammonium production and consumption in
cattail (Typha latifolia) marsh sediments. Using 15 N
techniques, he estimated the turnover time in this low
ammonium environment, at 13 ◦ C, to be 1–2 weeks.
Hemond (1983) used 15 N techniques to infer ammo-
nium sorption and nitrate reduction in Sphagnum wet-
lands. He found ammonium 15 N disappeared in batch
tests in about 10 days, with a much smaller correspond-
ing increase in Sphagnum 15 N.
Questions remain concerning the route by which
ammonium is converted to nitrous oxide and dinitrogen
in treatment wetlands (Tanner et al., 2002; Tanner and
Kadlec, 2003). The conventional assumption is:
NH4 → NO2 → NO3 → N2 O, N2
Under this scenario, the rates of ammonium removal
in the mesocosms would require a very large oxy-
gen supply, with no obvious source. Alternative less
 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381
oxygen-demanding pathways that operate in low oxy-
gen environments, such as anammox (Jetten et al.,
1999) may be involved, as recently positively identi-
fied in treatment wetlands (Shipin et al., 2004).
5. Conclusions
These studies have demonstrated that the details of
nitrogen processing in SSF wetlands include strong
interactions between water, sediment and biofilm
solids, and plant storages of nitrogen. Wetlands oper-
ating in a stable mode, past initial start-up phenomena,
were subjected to stable isotope impulse testing. Both
the mesocosm cascades and the field-scale wetlands
had similar water detention times of 4–5 day. However,
a 15 N ammonium impulse took 30–40 days to move
through the first 2–3 m of the field-scale wetlands and
∼120 days to exit the complete systems, and 25 days
was insufficient to recover more than 16% of the iso-
tope in the outflow from the mesocosm wetlands. The
interchange of nitrogen between sediments and water
has been established as the key delay mechanism, and
plant uptake and cycling as a secondary longer term
delay process.
Ammonium 15 N is rapidly sorbed into wetland
solids in the inlet region of the wetland, and subse-
quently gradually released in major part back to the
water. The next downstream elements of solids then
adsorb the release. This gradual “park and go” path
through the wetland has been termed “spiraling” in
the literature. Simultaneously, plant growth extracts
its nitrogen requirement from the system, quite likely
withdrawing it from the sorbed pool associated with
sediments and biofilms, that act as an important “park-
ing place” for nitrogen in transit. Calibration of a
two compartment model to the observed field-scale
response indicates solid phase nitrogen turnover rates
of about 1 day, suggesting that an atom of nitrogen
makes multiple stops (spirals) in its transit to the wet-
land outlet.
Nitrogen taken up into plant leaves and roots is sub-
ject to another type of delay, in which senescence and
decomposition are required before the nitrogen can
(possibly) find its way back to the water and sedi-
ments. This cycle has an average half life of over 1 year
for components of the plant used here (Schoenoplectus
tabernaemontani; Tanner, 2001).
379
Thus, it is seen that there are three characteristic
times that influence transit of ammoniacal nitrogen
in these SSF wetlands: the water detention time, the
sediment spiraling time, and the period of the biogeo-
chemical cycle. In a completely steady operating mode,
data analysis may proceed without regard for the details
of internal processes and delays. Traditional models,
such as the popular steady first order approach, may
be applied without concern for wetland dynamics. But
treatment wetlands may be subject to variable inputs
for two reasons: there may be stochastic or cyclical
variability of influent flows and/or nitrogen concentra-
tions, or the wetland inflow may be event-driven, as
for stormwater treatment applications. It may then be
advantageous to consider time averaging as a method
of avoiding complicated dynamic modeling, together
with the increased parameter numbers that unavoidably
accompany it.
Inert tracer testing has been employed in many SSF
studies, and demonstrates that there is a distribution
of detention times within the water column (Fisher,
1990; King et al., 1997). Wetlands similar to the field-
scale systems were tracer tested, and found to possess
broad residence time distributions (Tanner and Sukias,
1995; Tanner et al., 1998b). The mesocosms were tracer
tested, and found to behave like a series of five two-
tank systems (Tanner et al., 2002). The common theme
among such investigations is the requirement for two to
three nominal detentions to completely expel a tracer
impulse. Thus the outflow concentrations at any time
reflect some degree of past history extending back those
two to three nominal detentions. Comparison of con-
temporaneous input and output concentrations, there-
fore, becomes improper, as does computation of first
order rate constants from them. This dynamic synoptic
error may be avoided if inflow and outflow concentra-
tions are averaged over at least three to five nominal
detentions.
This study suggests that inlet disturbances in ammo-
nium concentrations may create another form of syn-
optic error, namely that caused by nitrogen spiraling.
In such an event, averaging would need to encompass
the distribution of solid-phase detention times as well
as the distribution of water-phase detention times. That
period would of necessity span nearly 1 year. During
such a long period, the seasonality of the biogeochem-
ical cycle (plant growth) would also come into play.
To encompass the biogeochemical cycle, averaging is
380 R.H. Kadlec et al. / Ecological Engineering 25 (2005) 365–381
required over several annual cycles. It is not reason-
able to expect output concentration patterns to “track”
inlet concentration patterns, as a water column model
would suggest. This is seen in the study of Tanner et al.
(1998a) where wetland outlet loads of NH4 -N showed
a 2–4 month delay in response to seasonal peaks in
influent loading.
This study also suggests that it could be very mis-
leading to average over the duration of a brief study
in which there are inflow variations, or to ignore plant
growth as a dynamic factor in a brief study. In lieu of the
commonly acceptable first order ammonium model, it
is far more preferable to utilize a three compartment
dynamic model that deals with storages in water, sedi-
ments, and plants.
Acknowledgements
This study benefited from funding by the New
Zealand Foundation for Research, Science and Tech-
nology, with collaboration facilitated by a grant from
NIWA’s Visiting Scientist Programme. Long Nguyen
and James Sukias of NIWA provided input to the design
and execution of the mesocosm studies, and Albert
van Oostrom helped in conception of these studies.
Results reported for small field-scale wetlands were
undertaken by VMH as part of a graduate research
program in the Biological Sciences Department at the
University of Waikato in collaboration with the Envi-
ronmental Management Section of the Meat Industry
Research Institute of New Zealand. This part of the
study benefited significantly from scientific input by
Prof. Warwick Silvester (UW), and Albert van Oost-
rom and John Russell (MIRINZ).
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