Wetlands Ecol Manage (2017) 25:649–666
DOI 10.1007/s11273-017-9542-2
ORIGINAL PAPER
An assessment of iron and calcium amendments
for managing phosphorus release from impacted Everglades
soils
Forrest E. Dierberg . Thomas A. DeBusk . Scott D. Jackson .
Michelle D. Kharbanda . Stacey C. Galloway . Patrick D. Owens .
Delia B. Ivanoff
Received: 5 July 2016 / Accepted: 14 March 2017 / Published online: 29 March 2017
 Springer Science+Business Media Dordrecht 2017
Abstract The recent implementation of agricultural
best management practices (BMPs) and treatment
wetlands called stormwater treatment areas (STAs)
have reduced phosphorus (P) concentrations and load-
ings to the Everglades Protection Area (EPA) in Florida
(USA). There is a concern that despite reductions in
external P loadings, internal loading from the legacy P
enrichment of the EPA wetland soils will continue to
elevate water column P concentrations, and may
impede restoration outcomes. In an effort to explore
ways to reduce soil P efflux, we retrieved intact,
vegetated (cattail, Typha domingensis) soil monoliths
from two P-enriched areas of the EPA and deployed
them at a location where they received pre-treated (low
P) surface water as ex situ flow-through mesocosms for
21 months with a mid-study 7-week dry down to
mimic natural hydroperiod conditions. Two treatments
Electronic supplementary material The online version of
this article (doi:10.1007/s11273-017-9542-2) contains supple-
mentary material, which is available to authorized users.
F. E. Dierberg (&)
T. A. DeBusk

S. D. Jackson
M. D. Kharbanda
S. C. Galloway

P. D. Owens
DB Environmental, Inc., 365 Gus Hipp Boulevard,
Rockledge, FL 32955, USA
e-mail: woody@dbenv.com
D. B. Ivanoff
Applied Sciences Bureau, South Florida Water
Management District, 3301 Gun Club Road,
West Palm Beach, FL 33406, USA
were tested for soils from both sites, using triplicate
mesocosms for each treatment. After applying a
herbicide (glyphosate) to eliminate the cattail vegeta-
tion, iron (Fe as liquid FeCl3) amendments provided no
P retention benefits in the organic soils from the two
sites, and did not yield significantly (P [ 0.05; n = 43)
lower flux rates (6.1 and 3.5 mg Pm-2 d-1) than the
herbicide/no soil amendment control (3.9 and 2.1 mg
Pm-2 d-1), as was expected. A combination of low
oxidation–reduction potential, heightened organic mat-
ter P mineralization, high pH, and sulfide production
acted interactively to enhance Fe and P mobilization in
the Fe-amended mesocosms. The herbicide/limerock
(CaCO3)-amended soils exhibited significantly lower
(P B 0.05; n = 43) P flux (1.3 and 1.1 mg Pm-2 d-1)
than the herbicide/no soil amendment control soils, but
it remains unknown whether the observed reduction in
P efflux (ranging from 48 to 67%) would justify the
expense and potential environmental impacts of apply-
ing a surficial limerock amendment to large regions of
the P-enriched wetlands.
Keywords Phosphorus
Immobilization
Iron

Limerock
Herbicide
Everglades
Introduction
A large portion of the Everglades Protection Area
(EPA) consists of Water Conservation Areas (WCAs),
which are shallow hydrology-managed wetland
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impoundments created during the mid-20th century
from a vast network of levees and canals in the
northern and central Everglades (Florida, USA). They
are characterized by marsh, slough, wet prairie, and
tree island habitats (Richardson 2008). The peat soils
in this region are underlain by calcitic mud, sandy clay
and sand, and a bedrock of Pleistocene limestone, and
typically exhibit circumneutral pH, high calcium (Ca)
content, and low iron (Fe) and aluminum (Al)
concentrations (Gleason et al. 1974).
Soil phosphorus (P) concentrations in interior (less
impacted) regions of the WCAs are typi-
cally B500 mg (kg dry wt)-1, but the historical
discharge of nutrient-enriched waters from the Ever-
glades Agricultural Area (EAA) and Lake Okee-
chobee to the WCAs has increased soil P in influent
regions, especially in WCA-2A, where TP levels have
exceeded 1500 mg (kg dry wt)-1 (Koch and Reddy
1992; DeBusk et al. 2001; Hagerthey et al. 2008). The
region of P enrichment in WCA-2A is estimated to
encompass 8100 ha. These areas now are colonized by
dense stands of cattail (Typha domingensis Pers.),
whereas WCA interior regions are characterized by
plants typical of oligotrophic conditions, including
sawgrass (Cladium jamaicense Crantz), spikerush
(Eleocharis cellulosa Torr.), and selected periphyton
(cyanobacteria: Scytonema hofmanii and Schizothrix
calcicola; hardwater diatoms: e.g., Mastogloia)
(McCormick and O’Dell 1996).
Most of the external water P loading that previously
contributed to elevated soil P levels in the WCAs is
now being curtailed with the operation of large,
constructed freshwater marshes called stormwater
treatment areas (STAs), as part of a long-term effort
to reduce P concentrations in agricultural drainage
water (ADW) and Lake Okeechobee surface water to
the Everglades (Chimney and Goforth 2001). Previous
investigations, however, have suggested that the
introduction of ‘‘low-P’’ waters into impacted regions
of the WCAs, and the resulting steep P concentration
gradient between the soil and water column, may
cause a significant and rapid flux of P from the
enriched soil to the water column (Fisher and Reddy
2001; Richardson et al. 2008).
Much of this P may be bioavailable, and could favor
continued, or even expanded, cattail proliferation
rather than the preferred sawgrass and periphyton
communities (Reeder and Davis 1983; Davis 1994;
Miao and DeBusk 1999). Consequently, wetland
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Wetlands Ecol Manage (2017) 25:649–666
Fig. 1 Location map depicting the locations (Sites N and C) c
where mesocosms were retrieved for the ex situ Intact Flow-
Through Soil Mesocosm Study (See Fig. A1 in Electronic
supplementary material for location of experimental site). The
F1, F2, and F4 locations depict where long-term ambient surface
water was collected and analyzed by United States Geological
Survey and South Florida Water Management District. The
‘‘Impacted’’ and ‘‘Moderately Impacted’’ zones shown on this
diagram were approximated from Koch and Reddy (1992)
restoration and recovery in the WCAs may be
prolonged due to the flux of P from enriched soil to
the water column.
The principal objective of this study was to evaluate
management practices to immobilize or reduce P in
the soil from impacted areas of WCA-2A. The
43,280 ha WCA-2A is one of the most nutrient-
impacted areas in the EPA, giving it a high priority for
restoration (Richardson 2008; Hagerthey et al. 2014).
This objective is related to the Long-Term Plan (Burns
& McDonnell Engineering Company Inc 2003) for
improving wetland regions considered to be impacted
by excess P.
Methods
Site description
The field collection sites for our study (Fig. 1) were
located within the northern, P-impacted region of
WCA-2A (Davis and Ogden 1994) dominated by
cattail. Both sites lie downstream from the S10-C
inflow structure, a point of historical discharge of
P-enriched water into WCA-2A via the Hillsboro
Canal. Site N is located near the northern extent of the
P-enrichment gradient, which extends in a general
southerly direction from S10-C; Site C is located
within the central region of the P gradient. The two
field sites are near sites of ongoing surface water
monitoring (Fig. 1). To date, several hundred acres of
cattail stands in WCA-2A near our study sites have
been controlled with fire and herbicides, and used for
both biogeochemical and ecological studies (Richard-
son 2008; Newman et al. 2013). Herbicide application
and fire also could be candidates for vegetation
reduction prior to the application of soil treatments,
such as chemical amendments.
Our study was conducted at the South Advanced
Technology Treatment Site (SATTS) adjacent to the
Wetlands Ecol Manage (2017) 25:649–666 651

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STA-1W outflow structure (Fig. A1; in Electronic
supplementary material). Outflow waters from STA-
1W contribute to the inflow waters to the S10-C inflow
structure, which is located approximately 30 km
downstream. Prior to the study, 22 intact soil mono-
liths (45.5-cm inside diameter and 40-cm deep) were
collected in cylindrical polyethylene containers (bar-
rels) from WCA-2A at Site N, in the vicinity of long-
term Everglades water quality monitoring station F1,
and at Site C (Fig. 1). The sub-regions within which
stations F1 and F3 are located have been previously
designated as ‘‘impacted’’ and ‘‘moderately impacted’’
(or ‘‘transitional’’), respectively, with regard to P
enrichment (Miao and Sklar 1998). A total of eleven
monoliths were retrieved from each site and trans-
ported to the SATTS research facility. All of the total
22 monoliths (henceforth referred to as ‘‘mesocosms’’)
initially contained intact cattail plants.
An inflow water pre-treatment (PT) system for the
mesocosms was implemented to reduce P concentra-
tions below that of ambient levels at field sites N and
C, and to mimic the inflow TP concentrations at the
S10-C structure (Fig. A2 in Electronic supplementary
material). Outflow water from STA-1W was treated in
two parallel pre-treatment trains, each consisting of
two 1600 gal tanks covered by water hyacinth
(Eichhornia crassipes (Mart.) Solms), followed by
six 1665 L (440 gal) tanks containing the submerged
aquatic plant southern naiad (Najas guadalupensis
(Spreng.) Magnus), before both trains were pumped to
a 1000 L (265 gal) mixing tank and then subsequently
routed to the mesocosms. Mean TP concentration in
the untreated STA-1W source water was 48 lg L-1
(n = 80), while TP in the treated water supplied to the
mesocosms averaged 22 lg L-1, during the study
period. A schematic of the design of a mesocosm is
presented in Fig. A1 (in Electronic supplementary
material).
We sourced pre-treated STA-1W water as inflow to
the mesocosms because conducting the study at the
remote in situ site at WCA-2A would not only have
been cost-prohibitive involving frequent use of air
boats and construction of solar panels, but would have
also compromised the quality of the results. This
would have included less frequent maintenance
required for adjusting flow rates, correcting power
outages, and flushing water pipes to clear bio-fouling.
Moving the soil monoliths to the STA-1W site, which
had electricity, provided easy access. Another
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Wetlands Ecol Manage (2017) 25:649–666
important factor in the decision to conduct the study
using ex situ mesocosms was the constraint imposed
by the natural seasonal dry down in WCA-2A, which
would have prevented access to both sites by airboat
due to low water depths. Furthermore, one of our focal
points was to examine the differences in P release from
impacted and moderately impacted soils. To accom-
plish this comparison, we elected to subject the soil
monoliths from the two sites to the same hydraulic
loading rate (HLR) and P loading under controlled
conditions throughout the 21-mo experiment. This
would not have been possible if the experiment had
been conducted in situ since the overlying water P
concentration would have been different between the
two sites (4 km distance) and over time.
Mesocosm treatments
The effectiveness of two chemical amendments was
examined using the flow-through mesocosms. Three
replicate mesocosms from each site were assigned the
following treatments:
• Herbicide application to eliminate cattails; no
amendment. This treatment was designated as the
control, since it is the precursor to application of
soil amendments.
• Herbicide application followed by soil surface
application at a thickness of ca. 1 cm (1560 g per
mesocosm or 9800 g m-2) of calcium carbonate
(CaCO3) as limerock (LR) consisting of a grain
size of fine sand (Fill #1 (DOT classification))
obtained from a nearby quarry (Palm Beach
Aggregates). The bulk density of the LR was
0.96 g cm-3. This treatment was designated as
‘‘Ca’’.
• Herbicide application followed by liquid ferric
chloride (FeCl3) applied by plant sprayer with
nebulizer to the surface soil at 100 g Fe(III) m-2
(Boers 1991; Smolders et al. 1995) followed by a
layer of sand (ca. 1 cm deep) to limit surficial
leaching of FeCl3 when flooded. This treatment
was designated as ‘‘Fe’’. A liquid form of iron
(FeCl3) rather than a solid Fe chemical (FeCl2,
FeCl3, FeSO4, Fe2O3, goethite [iron oxhydroxide,
FeOOH], or magnetite [Fe3O4]) was selected since
less studies in the literature applied liquid iron
amendments, which warranted further exploration
of the viability of this form of iron amendment for
Wetlands Ecol Manage (2017) 25:649–666
removing P. We assumed that the Fe(III) in the
liquid solution would initially be adsorbed onto
soil particles, and less likely to be desorbed during
the operational period by capping the soil with a
1 cm depth of sand.
The dates and the actions taken to prepare the
mesocosms for treatments and flow-through operation
can be found in the Electronic supplementary material.
After commencement of flow-through operations,
the mesocosms were maintained at a depth of 30 cm, a
mean HLR of 4.8 cm d-1 (range of 4.3–5.2 cm d-1),
and a flow-through mean hydraulic retention time
(HRT) of 6.3 days (range of 5.8–7.0 days) by daily
pulses of PT water. The mesocosms were drained after
one year in order to evaluate the effect of alternating
wet and dry seasonal cycles on P release. We initially
drained the mesocosms by siphoning the water to less
than 2 cm depth at 50 weeks after start of flow-
through operations. The mesocosm water was further
drained completely by drilling 3/8-inch holes in the
sides of the mesocosms. The dry down period, which
was imposed only once during the demonstration
period, lasted approximately seven weeks until rehy-
dration, followed by a return to normal flow-through
conditions (mean HLR = 4.8 cm d-1) three days
later.
Sampling protocol
Surface water
Surface water was sampled for total P (TP), and pH
and temperature were measured in situ on a bi-weekly
frequency. Samples for soluble reactive P (SRP) and
total soluble P (TSP) were collected monthly during
the first year, after which sampling was discontinued
for the scheduled dry down study. Samples were also
collected bimonthly for total alkalinity, dissolved Ca,
dissolved Fe, sulfate, and sulfide (the sum of all
dissolved sulfide species, H2S ? HS- ? S2-, here-
after referred to as sulfide) during the first year.
We began collecting surface water samples
(*2 cm below water surface) at the onset and after
3.5 days (*one-half HRT) from the resumption of
flow-through after the seven-week dry down for
analysis of TP, SRP, TSP, total alkalinity, dissolved
Ca, and dissolved Fe, and measuring temperature and
pH. Samples were also collected after 7 (*one HRT),
653
14 and 28 days. We continued sampling thereafter for
TP analysis, and measuring pH and temperature every
other week. In addition, samples were collected for
SRP, TSP, total alkalinity, dissolved Ca, dissolved Fe,
and sulfate on three occasions; sulfide was only
collected in only two of those three occasions.
Porewater
Porewaters were sampled periodically during the
study, and analyzed for TSP, SRP, dissolved Ca,
dissolved Fe, sulfide, and sulfate; pH and temperature
were measured in situ. The first event was prior to the
start of the study, when two mesocosms from each of
Sites N and C were sacrificed for soil characterization.
The 0–10 and 10–30 cm soil depths were retrieved
from each mesocosm after collecting the loose cattail
leaf litter. Each soil layer from each site was compos-
ited, and then subsampled after homogenizing the soil,
and later centrifuged in the lab to retrieve porewater.
Thereafter, porewater was sampled from a soil depth
of 6–10 cm below the soil surface by use of a ‘‘sipper’’
(Stober et al. 2001). The first was after herbicide
application but prior to chemical amendments; the
second occurred after 50 weeks of flow-through
operation; and the last two took place 3.5 and
8 months after resumption of flow-through following
the seven-week dry down. See Electronic supplemen-
tary material for the exact dates of porewater sampling
by ‘‘sipper’’. We also measured oxidation–reduction
potential (ORP) at mid-water column and in the
porewater at a depth of 6–10 cm below soil surface on
all four occasions.
Soil
Two soil depths, 0–10 and 10–30 cm, were sampled
on December 13, 2006 for initial soil characterization
from each of the two sacrificed intact soil cores
retrieved from each of the field sites. We analyzed the
soils for bulk density, moisture content, TP, and
inorganic P fractions. For each control and treatment
group at the end of the 21-month experiment, total P
was analyzed on the 0–10 cm depth soil for all three
replicates while inorganic P fractionation was per-
formed for two of the three replicate mesocosm soils.
For the 10–30 cm soil depth, TP was again measured
on all three replicates, but inorganic P fractionation
was performed on only one of the mesocosm
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replicates. These analyses provided data on the
relative differences between labile and nonlabile P
fractions in the ‘‘impacted’’ (Site N) and ‘‘moderately
impacted’’ (Site C) soils.
Comparison of soil TP concentration among treat-
ments was made on a mass/volume basis (lg P cm-3)
rather than a mass/mass basis (lg P g-1), because of
differences in soil bulk density among treatments
imposed by the addition of LR and sand (to cap the
FeCl3 amendment). The bulk density of the LR was
0.96 g cm-3, which increased the densities of the Site
N and Site C soils that received the amendment. The
1.0 cm thick layers of LR and sand added to the soil
surface also ‘‘diluted’’ the soil P in the 0–10 cm depth
horizon since the LR and sand contained negligible P
concentrations.
Analytical methods
Filtered surface water and porewater samples were
analyzed according to SM 4500-P F (APHA et al.
1992) for TSP following digestion, and for SRP on
undigested samples. Unfiltered surface water samples
were analyzed for TP via SM 4500-P F after digestion.
Porewater dissolved organic P (DOP) concentration
was calculated by the difference between the mea-
sured TSP and SRP concentrations; the difference
between TP and TSP concentrations yielded surface
water particulate P (PP). Sulfate was analyzed accord-
ing to SM 4110B (APHA et al. 1992) or EPA 375.4
(USEPA 1999), while sulfide was measured with an
ion-selective electrode (ISE) according to a modifica-
tion of the SM 4500-S2- G method (APHA et al.
1998). Dissolved Fe was determined using a modified
bathophenanthroline procedure (Nürnberg 1984). Dis-
solved Ca and total alkalinity were measured accord-
ing to EPA methods 215.1 and 310.1, respectively
(USEPA 1999).
Bulk density and moisture content were determined
according to ASA 13 and ASA 21-2 methods (ASA
1986). Total P was analyzed using EPA 365.2
(USEPA 1999) after perchloric acid digestion (Plumb
Jr. 1981). Soil analysis for inorganic P fractionation
was performed following Reddy et al. (1998). As a
quality assurance check, we found good agreement
between the measured residual P concentrations that
remained after the sequential inorganic P extraction
steps and calculated residual P, which is the difference
between the initial soil TP and the sum of the extracted
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Wetlands Ecol Manage (2017) 25:649–666
P fractions (Fig. A3; in Electronic supplementary
material).
The Florida Department of Environmental Protec-
tion (FDEP) standard operating procedures for field
Activities (FDEP 2004) was used in the analysis of all
field parameters: pH (FT 1100), temperature (FT
1400), specific conductivity (FT 1200), and dissolved
oxygen (FT 1500). The ORP was measured with a
Hach Platinum (Pt) Series ORP electrode (Part no.
51937-00) and a Hach sensION2 pH/ISE meter.
Calibration of the Pt electrode (referenced to an Ag/
AgCl electrode in saturated KCl) was accomplished
by measuring the potential of a Light’s solution (Light
1972). Electrode potentials of the porewaters were
subsequently adjusted to a standard hydrogen elec-
trode (SHE) and for deviations in temperature from 25
C (APHA et al. 1998).
Data analysis
Statistica version 9.1 (Statsoft, Inc. 2010) was used to
execute statistical analyses for determining signifi-
cance. Either an independent t test or a one-way
ANOVA using the Tukey HSD for post hoc compar-
isons was used to test significance of differences
between the means (comparisons between sampling
sites and among treatments) when the data were
normally distributed and had homogeneity of vari-
ances. The Shapiro–Wilk and Bartlett Chi Square tests
were used to determine if the data were normally
distributed and the variances were homogeneous,
respectively. Whenever the test for homogeneity
failed, we plotted the mean versus the standard
deviation, and if the plotted data were correlated, then
the data were transformed. In the instance when either
normality or homogeneity were not met, log or square
root transformations were often successful in convert-
ing the data to meet normality or homogeneity
assumptions. In the instance when the data could not
be successfully transformed, the non-parametric
Mann–Whitney U-test (for comparing two groups)
or Kruskal–Wallis ANOVA (for comparing multiple
groups) by Ranks with a post hoc multiple comparison
of the mean ranks was used.
Comparisons of surface- and porewater concentra-
tions and P fluxes between discrete phases of the study
(e.g., pre-dry down, reflood [two-week period follow-
ing dry down], and post-dry down (remainder of
sampling period)] were performed using t-test or
Wetlands Ecol Manage (2017) 25:649–666 655

nonparametric Wilcoxon Rank-Sum or Mann–Whit-
ney U-test following the same criteria for meeting the
assumptions of the test as outlined above. Significance
was determined using either one-tailed (in the case of
treatment versus control; or Site N vs. Site C) or two-
tailed (in the case of comparing the two treatments or
pre-vs. post-dry down) testing with a = 0.05.
Results
Surface water outflow
Both soil amendment treatments and the herbicide-
only control tended to export P relative to the low-P PT
inflow water (Fig. 2) which contained a mean TP
concentration of 22 lg L-1 (Table 1). Mesocosms
retrieved from Site N generally had higher mean
Inflow Site N Site C
350
CT
300
Dry
250
TP (µg L-1)
surface water TP concentrations than those from Site
C for the same treatment, with concentrations being
significantly (P B 0.00001; n = 45; one-tail) higher
in Site N mesocosms for the control and Fe treatment.
Between the two soil amendments (LR and FeCl3),
LR inhibited soil P release to a greater extent than did
FeCl3, for both soil types (P \ 0.0001; n = 45; two-
tailed). Addition of LR also reduced outflow TP
concentrations to levels that were significantly less
(P \ 0.02; n = 45; one-tailed) than the Control
mesocosms (herbicide without soil amendment) at
Site N and Site C (Table 1; Fig. 2). On the other hand,
Fe mesocosms from Site N and Site C did not produce
significantly lower (P [ 0.98; n = 45; one-tailed)
outflow P concentrations than the Control mesocosms,
although outflow P concentrations decreased towards
the end of study period (Fig. 2).
Aside from a short-lived post-dry down ‘‘first
flush’’, outflow SRP concentrations for the two
treatments and Control were typically below 21 lg
L-1 (Fig. A4; in Electronic supplementary material).
Nevertheless, the mean outflow SRP concentrations

Down
200
for the entire study period (including the ‘‘first flush’’
150
after the dry down) ranged from 6–28 lg L-1 for the
100 Control, Fe, and Ca mesocosms, which were higher
50 than the mean inflow SRP concentration of 3 lg L-1
0 (Table 1). The Fe mesocosms from both collection
sites exhibited variable SRP concentrations through-
350
Ca out the study.
300
Particulate P and DOP species constituted the
TP (µg L-1)

250 Dry
200 Down majority of total P in the outflow of all mesocosms
150 (Table 1), with Fe mesocosms averaging the highest
100 outflow PP concentrations from both sites [means for
50 Site N = 94 lg L-1 (P B 0.0001 and P B 0.004; two-
0 tailed) and Site C = 41 lg L-1 [P = 0.02 and 0.125;
350
two-tailed) for comparisons with LR and Control
Fe mesocosms, respectively]. Although not statistically
300 Dry
Down significant, the Control mesocosms had the highest
TP (µg L-1)

250
200 mean outflow DOP concentrations among all meso-
150 cosms at both sites (means for Site N = 42 lg L-1 and
100 Site C = 28 lg L-1). Compared to SRP, the export of
50 DOP and PP upon reflooding the mesocosms after the
0
A M J J A S O N D J F M A M J J A S O N D J
seven-week dry down was suppressed except for the
2007-2009 DOP from Control and PP from Fe treatments of the N
soils (Fig. A4; in Electronic supplementary material).
Fig. 2 Mean (±1 SE; n = 3) total phosphorus (TP) concen- As with the P concentrations, dissolved Ca and Fe,
trations in the inflow and outflow of the mesocosms containing
sulfate, and sulfide concentrations increased in the
soil from the N and C field sites. CT: herbicide applied to
eliminate cattail (control), Ca: herbicide followed by limerock outflows of all Control and treatment mesocosms
amendment, Fe: herbicide followed by FeCl3 amendment compared with the inflow concentrations; alkalinity

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Table 1 Mean (±1 SE) outflow concentrations during the demonstration period from intact flow-through soil mesocosms retrieved
from Sites C and N in WCA-2A and subjected to chemical amendments after herbicide application
Parameter Inflow Site N Site C
CT Ca Fe CT Ca Fea

TP (lg L-1) n = 45 22 ± 1.3 107 ± 8.0 51 ± 4.1 146 ± 9.7 67 ± 6.0 46 ± 2.6 94 ± 5.8
SRP (lg L-1) n = 17 3 ± 0.4 28 ± 8.6 6 ± 1.5 20 ± 5.4 11 ± 4.6 7 ± 2.1 16 ± 3.9
DOP (lg L-1) n = 17 8 ± 0.8 42 ± 11.7 25 ± 7.6 33 ± 4.7 28 ± 8.1 17 ± 3.1 26 ± 3.6
PP (lg L-1) n = 17 10 ± 1.2 50 ± 6.2 30 ± 4.2 94 ± 12.6 36 ± 5.6 29 ± 3.2 41 ± 2.0
pH n = 44 8.11 ± 0.03 8.03 ± 0.04 8.26 ± 0.07 8.09 ± 0.04 8.03 ± 0.06 8.22 ± 0.05 7.95 ± 0.05
Alkalinity (mg L-1) n = 13 206 ± 6.7 211 ± 11.9 204 ± 11.9 214 ± 9.3 207 ± 12.3 206 ± 8.8 186 ± 11.2
Ca (mg L-1) n = 13 55 ± 3.0 63 ± 3.9 57 ± 4.0 70 ± 6.1 62 ± 4.0 60 ± 3.7 70 ± 7.2
Fe (lg L-1) n = 12 45 ± 3.3 46 ± 5.1 42 ± 2.6 254 ± 111 42 ± 4.2 47 ± 6.3 253 ± 55
Sulfate (mg L-1) n = 13 37 ± 4.3 59 ± 9.8 53 ± 8.0 63 ± 14.0 59 ± 9.5 61 ± 9.3 75 ± 19.2
Sulfide (lg L-1) n = 9 26 ± 2.9 42 ± 5.0 42 ± 2.7 33 ± 2.8 31 ± 3.8 31 ± 2.7 27 ± 2.3
ORP (mV) n = 3 260 ± 23.8 254 ± 30.9 266 ± 28.4 267 ± 25.9 270 ± 26.4 275 ± 25.4
a
Mean represents n = 3 until May 30, 2008; thereafter represents n = 2. Time-weighted standard error of three replicates through
May 30, 2008 and of the two replicates after May 30, 2008
n = number of samples analyzed for the inflow and each treatment mesocosm at each site. The duration of the demonstration was
21 months and included a seven-week dry down period
CT: herbicide applied to eliminate cattail (control), Ca: herbicide followed by limerock amendment, Fe: herbicide followed by FeCl3
amendment

levels remained about the same (Table 1). Due to the
Fe(III) treatment, outflow dissolved Fe concentrations
were 5–6 times higher than the inflow concentration for
both soils of the Fe mesocosms, and at least 4 times
higher than any of the other Control or treated meso-
cosm outflow concentrations. Nearly all of the Fe
release in the FeCl3-amended mesocosms occurred
during the first seven months prior to the dry down, and
then again, but to a lesser extent, during the four months
post-dry down. Moreover, the pH values of the outflow
waters from Sites N (7.59–7.80) and C (7.45–7.74) were
lower during the first two months after applying the
FeCl3; the pH was also lower in the outflows from both
sites immediately upon reflooding after the dry down
(Fig. A5; in Electronic supplementary material). These
pH values, which were lower than those measured in the
outflows from the Control and Ca-treated mesocosms
(data not shown), were likely due to the acidity produced
from the hydrolysis of the Fe within the upper soil that
received the FeCl3 amendment. The lower pH in the
Fe(III)-amended mesocosms contributed to the higher
dissolved Ca concentrations than in the Control and Ca-
treated mesocosms (Table 1). The dissolved Ca con-
centrations in outflows of the Ca treatments were within
the range of the Control mesocosms, indicating that
unlike the applied FeCl3, the LR was stable during the
experiment. The highest water column pH values were
associated with the Ca treatment for both soils, due to the
buffering capacity of the added LR.
During the first two weeks of reflooding follow-
ing the seven-week dry down, mesocosms from both
sites exhibited an initial increase in water column
TP concentrations, relative to pre-dry down TP
concentrations (Fig. 2). SRP was also mobilized in
the control and both soil treatments (Fig. A4; in
Electronic supplementary material). However, as
with TP, the releases in all mesocosms were short-
lived, with SRP concentrations returning to their
pre-dry down levels within two weeks (*two
HRTs) after reflooding. Further results of the dry
down-reflood manipulation can be found in the
Electronic supplementary material.
Porewater
The mean porewater pH ranged from 6.85 to 7.54
among all the mesocosms and sampling events for
both sites (Table 2). Lower ORP (Eh) across all

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Wetlands Ecol Manage (2017) 25:649–666 657

Table 2 Mean (±1 S.E.) concentrations of physico-chemical parameters in porewaters (6–10 cm soil depth) from the ex situ intact
flow-through soil mesocosms for each sampling event
Parameter Sampling event Site N Site C
CT Ca Fe CT Ca Fe

pH (units) Post-herbicide 7.33 7.40 7.54 7.33 7.39 6.95

Initial: Pre-dry down 7.49 ± 0.13 7.48 ± 0.12 7.54 ± 0.06 7.13 ± 0.12 7.02 ± 0.06 6.85 ± 0.12
Site C = 7.49 Post-dry down 7.49 ± 0.07 7.37 ± 0.06 7.41 ± 0.05 7.43 ± 0.09 7.17 ± 0.02 7.25a ± 0.04
Site N = 7.75 Final 7.29 ± 0.04 7.26 ± 0.12 7.24 ± 0.07 7.37 ± 0.06 7.25 ± 0.14 7.31a ± 0.05
Eh (mV) Post-herbicide -15 -85 -71 -45 -57 -44
Pre- dry down 9 ± 20 -30 ± 11 6 ± 16 22 ± 11 41 ± 14 -87 ± 77
Post- dry down -141 ± 15 -158 ± 5 -152 ± 2 -155 ± 10 -121 ± 19 -142a ± 2
Final -95 ± 25 -109 ± 11 -136 ± 36 -117 ± 1 -120 ± 9 -98a ± 1
-1
Sulfate (mg L ) Post-herbicide 19.0 34.0 31.0 20.0 25.0 14.0
Initial: Pre-dry down 11.4 ± 4.8 8.5 ± 2.8 3.8 ± 1.1 2.8 ± 1.5 9.9 ± 1.2 2.7 ± 0.9
Site C = 15.4 Post- dry down 2.9 ± 1.9 8.1 ± 3.8 7.0 ± 2.1 8.6 ± 2.8 10.6 ± 3.9 5.2a ± 2.1
Site N = 49.5 Final 12.8 ± 5.3 17.2 ± 8.2 21.0 ± 7.5 15.5 ± 5.9 8.4 ± 6.3 12.0a ± 6.1
Sulfide (mg L-1) Post-herbicide 5.3 2.6 0.7 3.9 3.6 5.8
Initial: Pre-dry down 2.8 ± 1.0 3.7 ± 0.7 1.6 ± 0.6 4.1 ± 0.2 3.3 ± 0.6 0.8 ± 0.3
Site C = 0.26 Post-dry down 3.3 ± 1.1 5.3 ± 0.1 1.3 ± 0.7 4.7 ± 1.7 4.8 ± 1.6 1.0a ± 0.8
Site N = 0.40 Final 4.3 ± 0.6 5.0 ± 1.4 2.9 ± 1.1 4.5 ± 1.4 5.5 ± 0.8 3.2a ± 0.0
-1
Diss. Fe (lg L ) Post-herbicide 37 16 16 67 75 90
Pre-dry down 44 ± 7 34 ± 4 128 ± 56 41 ± 4 40 ± 4 1343 ± 987
Post-dry down 49a ± 11 215 ± 109 307 ± 184 76 ± 19 64 ± 15 195a ± 112
Final 35 ± 13 25 ± 12 97 ± 55 13 ± 0 23 ± 10 81a ± 31
Initial values (n = 2) for Site N and Site C soils (0–10 cm depth) are provided where available
a
n=2
n = 3 except for the post-herbicide (prior to soil amendments) sampling event (n = 1), and where otherwise noted. The duration of
the demonstration was 21 months and included a seven-week dry down period
CT: herbicide applied to eliminate cattail (control), Ca: herbicide followed by limerock amendment, Fe: herbicide followed by FeCl3
amendment

controls and treatments at both sites occurred during
the two porewater samplings after reflooding the
7-week dry out soils in the mesocoms. Sulfate levels
decreased from their initial and post-herbicide appli-
cation concentrations during pre- and post-dry down
periods, but then increased across the control and
treatments for both sites at the end of the 21 months
(Table 2). Sulfide concentrations increased from ini-
tial values of 0.40 (Site N) and 0.26 (Site C) mg L-1
after initially flooding the mesocosms, and usually
remained elevated (typically [ 1.0 mg L-1) through-
out the 21-month experimental period. There were no
obvious sulfide concentration differences between
sites, but the Fe-treated mesocosms exhibited lower
sulfide levels than did the Control or Ca-treated
mesocosms for both sites. Due to the FeCl3 amend-
ment that took place subsequent to the post-herbicide
sampling event, porewater dissolved Fe concentra-
tions in those treated mesocosms increased substan-
tially compared to the other mesocosms (Table 2),
with the mesocosms from Site C exhibiting higher
dissolved Fe concentrations compared to those at Site
N through the pre-dry down sampling.
Prior to herbicide application or chemical amend-
ments, mean soil porewater (0-10 cm soil depth) SRP
and DOP concentrations were higher in the Site N soil
(153 lg SRP L-1 and 89 lg DOP L-1) than the Site C
soil (29 lg SRP L-1 and 43 lg DOP L-1). Two
months later (post-herbicide for control, Ca, and Fe
treatments but prior to soil amendments), under batch

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658 Wetlands Ecol Manage (2017) 25:649–666

feed conditions, porewater (6–10 cm below soil
surface) SRP concentrations in the Site N soils were
reduced relative to the initial concentration (Fig. 3). In
contrast, porewater SRP concentrations in mesocosms
from Site C were higher than the initial 29 lg L-1 for
those treatments.
The elevated DOP concentrations in the porewater
immediately after herbicide application decreased
dramatically in the Control and two amended meso-
cosms from both sites during the subsequent 49 weeks
of flow-through operation prior to dry down (Fig. 3).
The addition of LR and FeCl3 did not reduce either
SRP or DOP porewater concentrations relative to the
Control mesocosms. However, the mean SRP concen-
tration was significantly lower (P = 0.025; n = 3;
two-tailed) for the Ca than Fe treated Site N soils after
49 weeks of flow-through operation during the pre-dry
down period.
For the two post-reflood porewater sampling events
(P-R and Final), SRP and DOP concentrations tended
to be higher for Site N than for Site C in the Control
mesocosms, and higher in SRP concentration for Site
C than for Site N in the Ca and Fe mesocosms (Fig. 3).
The first post-reflood porewater sampling, 15 weeks
after the resumption of flow, demonstrated slightly
higher SRP and DOP concentrations than levels
observed prior to dry down for the control and
treatments from Site N (Fig. 3). A greater variability
in SRP and DOP concentrations were exhibited in the
Site C mesocosm porewaters.
By the last collection date, 35 weeks post-reflood
after the dry down, porewater SRP and DOP

Site N Site C Site N Site C

200 1200 n=1
CT CT
1000
150
SRP (µg L-1)

n=1
DOP (µg L-1)

800
n=1
100 600

n=1 400
50
200
0 0

200 1200
Ca Ca
1000
150
DOP (µg L-1)
SRP (µg L-1)

800 n=1

100 n=1 600

n=1
n=1 400
50
200
0 0

n=1 n=1
200 1200
Fe Fe
1000
150
DOP (µg L-1)
SRP (µg L-1)

n=2 800
n=2 n=1
100 600
n=2
n=1 400
50
n=2 n=2
200 n=2
0 0
P-H P-D P-R Final P-H P-D P-R Final

Fig. 3 Mean (±1 SE; n = 3 except where noted) soluble
reactive phosphorus (SRP) and dissolved organic phosphorus
(DOP) concentrations of the porewaters from the ex situ intact
flow-through soil mesocosms. Initial porewater concentrations
for soils at Sites N and C (0–10 cm depth) are indicated by long-
and short-dashed lines, respectively. All other porewater
concentrations were collected from a soil depth of 6–10 cm
below soil surface. CT: herbicide applied to eliminate cattail
(control), Ca: herbicide followed by limerock amendment, Fe:
herbicide followed by FeCl3 amendment. P–H: Post-Herbicide
but prior to soil amendments; P–D: Pre-Dry Down; P–R: Post-
Reflood

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Wetlands Ecol Manage (2017) 25:649–666 659

concentrations in all mesocosm Control and treat-
ments for both soils had decreased relative to levels
measured 15 weeks following the dry down-reflood
event (Fig. 3). The Ca treatment continued to exhibit
lower porewater SRP and DOP levels than the Fe
treatment, but as in the three previous porewater
sampling dates, the differences generally were not
significant.
Vegetation composition
Composition of the vegetation within each mesocosm
was noted throughout the 21-mo study. See Table A1
(in Electronic supplementary material) for a summary
of the periods before and after the 7-week dry down.
Soil characteristics
Site N soil, collected in the ‘‘impacted’’ region, was
initially more P enriched (79.3 lg cm-3, or 1485 mg
(kg dry wt)-1) than the ‘‘moderately impacted’’ Site C
soil (68.9 lg cm-3, or 1085 mg (kg dry wt)-1) in the
upper 10 cm depth interval (Fig. 4). For the 10–30 cm
soil depth, the difference between Sites N and C soils
140
Depth: 0-10 cm
120
TP (µg cm-3)
was even greater: 74.4 lg cm-3 versus 46.0 lg cm-3
(1380 vs. 596 mg (kg dry wt)-1), respectively.
The initial total organic carbon (TOC: 47.3%), total
nitrogen (N: 3.04%), total Ca (3.1%), and total Fe
(0.12%) concentrations for the 0–30 cm depth interval
for the N site were similar to those at the C site soils
(47.8% TOC, 3.09% N, 2.6% Ca, and 0.11% Fe).
These constituents were also similar between soil
depths of 0–10 cm versus 10–30 cm within a site (data
not shown), except for Fe at Site C where the
10–30 cm soil layer was 77% more enriched with Fe
than the 0–10 cm layer (0.13 vs. 0.07%).
In both the 0–10 and 10–30 cm soil layers, mean TP
concentration in the Ca mesocosms from both sites
was higher at the end of the study than at the beginning
of the study (prior to imposing treatments) (Fig. 4).
This was also true for the 10–30 cm soil depth when
compared to the Control, but not for the 0-10 cm soil
depth interval. The Fe-amended soil TP concentra-
tions at the end of the 21-mo study more closely
approximated the initial and Control (end of study) TP
concentrations across both soil depth intervals and
locations.
The inorganic P fractionation provided insight into
which P pools were most affected by the treatments,
and whether they were labile or recalcitrant. Within
the more labile P pools (readily available and Fe/Al-
bound), P was usually approximately equal to or
higher in all chemically-treated soils (including the

100
80
herbicide-only control) compared to the initial (pre-
60 treatment) soils from both sites (Fig. 5). Notable ex-
n=2
40 ceptions include the low readily available P in the Fe-
20
0
amended mesocosms for the 0–10 cm soils at Sites N
140
and C.
Depth: 10-30 cm
120 For the more nonlabile biogenic and Ca/Mg-bound
TP (µg cm-3)

100 P fractions in the Control, and Fe and Ca treatments,

80
n=2 concentrations did not generally vary meaningfully
60
40 after 21 months from the initial values measured prior
20
0
to treatment, within either the 0–10 (P [ 0.62; n = 3;
Initial CT Ca Fe Initial CT Ca Fe two-tailed) or 10–30 cm soil depths (Fig. 5). Excep-
End of Study End of Study tions were found in the Ca/Mg-bound P pool, where
Site N Site C the Ca-treated 0-10 cm soil from both sites and the Fe-
amended 10–30 cm soil from Site N were elevated.
Fig. 4 Mean (±1 SE; n = 3) total phosphorus (TP) concen-
trations in the 0–10 and 10–30 cm soil depth intervals of Between the two sites, the biogenic and Ca/Mg-bound
mesocosms from the N and C field sites in WCA-2A at the P pools were lower in the 10–30 cm soil depths from
beginning (initial—December 13, 2006, prior to treatments with Site C than for Site N soils across all treatments; this
herbicide and soil amendments) and end of the study (January
was not true for the 0–10 cm soil depths, where similar
13, 2009). CT: herbicide applied to eliminate cattail (control),
Ca: herbicide followed by limerock amendment, Fe: herbicide concentrations were observed across all treatments
followed by FeCl3 amendment (Fig. 5).

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660 Wetlands Ecol Manage (2017) 25:649–666

Depth: 0-10 cm Depth: 10-30 cm

Readily Available
1.2 1.2

P (µg cm-3)
0.8 0.8

0.4 0.4

0.0 0.0

5 5
Fe/Al-bound P

4 4
(µg cm-3)

3 3
2 2
1 1
0 0

30 30
Biogenic P
(µg cm-3)

20 20

10 10

0 0
60 60
Ca/Mg-bound P
(µg cm-3)

40 40

20 20

0 0
CT
CT

Fe

Fe
Ca

Ca
Initial

Initial

CT

Fe
Ca

CT

Ca

Fe
Initial

Initial

End of Study End of Study End of Study End of Study

Site N Site C Site N Site C

Fig. 5 Readily available, biogenic, Fe/Al- and Ca/Mg-bound
phosphorus (P) concentrations in the 0–10 cm (n = 2) and
10–30 cm (n = 1 except for initial (n = 2)) soil depth intervals
of the ex situ intact flow-through mesocosms from N and C field
sites before and after 21 months of treatment. CT: herbicide
applied to eliminate cattail (control), Ca: herbicide followed by
limerock (LR) amendment, Fe: herbicide followed by FeCl3
amendment. The initial represents P concentrations on Decem-
ber 13, 2006 prior to treatments with herbicide and soil
amendments (LR and Fe salt). Error bars represent ±1 SE Note
the different Y-axis scales

Phosphorus flux rates
Among the mesocosm treatments (Ca and Fe) and
Control, the soils retrieved from Site N released
more P than the Site C soils (Table 3), indicating a
larger labile P pool in the more highly impacted Site
N soil. Application of Fe to the surface soil after
herbicide treatment (Fe treatment) sharply exacer-
bated the release of soil P relative to the Control
mesocosms from both sites, with rates of 6.1 and
3.5 mg P m-2 d-1 observed for the Site N and Site
C soils, respectively. The next highest release rates
were 3.9 (Site N) and 2.1 (Site C) mg P m-2 d-1 for
the Control mesocosms, which were treated with
herbicide but no soil amendments. The LR amend-
ment following herbicide application (Ca treatment)
apparently was successful in inhibiting soil P
release, as the mean P flux for the Ca treatment
was lower (by more than 48%) than in the Control
mesocosms (Table 3).

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Wetlands Ecol Manage (2017) 25:649–666 661

Table 3 Mean (±1 SE) total phosphorus (TP) flux rates from during the next 39 days before increasing again
intact flow-through soil mesocosms retrieved from Site C and (Fig. 2). The post-dry down period corresponded to
Site N in WCA-2A and subjected to the following treatments
lower flux rates than those measured during the pre-
Site/Treatment P Flux (mg P m-2 d-1) dry down period, averaging 3.0 and 1.2 mg P m-2 d-1
Mean SE versus the pre-dry down values of 4.6 and 2.9 mg P
m-2 d-1 for the Site N and Site C soils, respectively.
Site N The statistically significant (P B 0.0017; n = 24 (pre-
CT 3.9 0.31 dry down) or 19 (post-dry down); two-tailed) decrease
Ca 1.3 0.15 in P flux during the latter part of the study compared to
Fe 6.1 0.43 the pre-dry down period can likely be attributed to the
Site C more rapid leaching and decomposition of highly
CT 2.1 0.26 labile plant detritus immediately after applying the
Ca 1.1 0.10 herbicide (DeBusk and Reddy 2005) during the onset
Fea 3.5 0.24 of the pre-dry down period. The 13.6% P contained in
a
Mean represents n = 3 until May 30, 2008; thereafter glyphosate likely also contributed to some of the
represents n = 2. Time-weighted standard error of three exported DOP initially during flow-through
replicates through May 30, 2008 and of the two replicates operations.
after May 30, 2008
The duration of the demonstration was 21 months and included Soil chemical amendments
a seven-week dry down period
CT: herbicide applied to eliminate cattail (control), Ca:
herbicide followed by limerock amendment, Fe: herbicide
For this study, we chose to investigate the potential
followed by FeCl3 amendment attenuation of P using Fe- and Ca-based chemical
amendments. Although alum (aluminum sulfate [Al2(-
SO4)3
18H2O] has been found to be a strong binding
agent in reducing P release from muck farmland in
Discussion central Florida (Hoge et al. 2003), alum was not
considered in our study due to concerns about
Herbicide application potential adverse effects of aluminum and sulfate ions
on Everglades biota (Sherwood and Qualls 2001). We
Research into cattail removal in the Everglades has applied the two chemical amendments (LR and FeCl3)
focused on prescribed burning, physical control on top of the soil after eradicating the cattails with
(through a combination of cutting and flooding), and herbicide, rather than incorporated into the soil, so as
chemical control (Richardson 2008; Hagerthey et al. to minimize upward P release from the soil to the
2014). These efforts have been made as precursors to water column (Chimney et al. 2007). Broadcasting
habitat restoration, and not necessarily as a forerunner chemicals on the surface maintains their high P
for managing P as is the intent of our investigation. For sorption capacity with respect to the flux of porewater
example, the removal of dense cattail stands in the SRP and P released from decomposition of organic
P-impacted areas of WCA-2A is a necessity if soil matter underneath.
amendments are to be effective in binding soil P. We We did not compare various application amounts
therefore used glyphosate herbicide to remove the within or between the two treatments since the study
standing cattail biomass. was conceived as a preliminary feasibility study to
In the herbicide-only Control mesocosms we determine whether either of the two chemical amend-
observed substantial releases of TP to the water ments deserved further consideration in future inves-
column prior to dry down (Fig. 2), especially in the tigations. Thus we did not seek to ‘‘normalize’’ the
DOP component during the first two months (Fig. A4; application amounts in terms of adsorption capacity,
in Electronic Supplementary material). P flux rates in and therefore a comparison in the effectiveness of P
these same mesocosms increased again immediately removal by each of the added substrates is limited to
upon reflooding following the dry down period, but P the specific application amounts employed in this
flux in the Control mesocosms subsequently decreased study.

123
662
Limerock
Various solid Ca-containing substrates such as cal-
cium silicate slag (wollastonite: CaSiO3) and calcium
carbonate (CaCO3) have been documented to adsorb,
precipitate, or filter P in Everglades soils at different
experimental scales: bench-top sorption/desorption
isotherms (Chimney et al. 2007), re-packed soil lab
core run in batch mode (Chimney et al. 2007), and
flow-through outdoor mesocosms (DeBusk et al. 2004;
this study). We chose LR rather than CaSiO3 because
LR is readily abundant in south Florida from quarrying
operations and lacks heavy metals that may be present
in CaSiO3 slag (Proctor et al. 2000). Also LR is a
prominent geological feature, underlying the shallow
soils, throughout most of the Everglades marshes.
LR was effective in inhibiting P release by factors
of 3.0 and 1.9 relative to the control treatments for N
and C site soils, respectively (Table 3). This was
verified by the considerably higher TP and Ca/Mg-
bound P concentrations in the 0–10 cm soil depth of
the LR-treated (Ca) mesocosms from both Sites N and
C at the end of the study, compared to either the initial
(prior to treatment) or Control mesocosms from those
sites (Figs. 4, 5). The Ca/Mg-bound P is generally
considered to be chemically and biologically recalci-
trant (White et al. 2004; Reddy and DeLaune 2008).
The effect of adding LR to the surface of the soil did
not have any effect on the TP (Fig. 4) or Ca/Mg-bound
P (Fig. 5) in the 10–30 cm soil depth interval when
compared to the initial or Control mesocosms.
Chimney et al. (2007) applied calcium silicate
(CaSiO3) slag generated during stainless steel produc-
tion to relict agricultural soils from one of the STAs.
They reported a 74 and 84% reduction in P flux for
application rates of 400 and 1000 g m-2 (4 and 10 t
ha-1), respectively, relative to the unamended control.
Although these percent reductions are high, the
corresponding P flux rates of 6.1 and 3.7 mg m-2
d-1 were approximately the same as the results from
Site N Control and both sites N and C for Fe treatment
in our study. However, the P release rates for the
herbicide Control mesocosms in our study (N site: 3.9
and C site: 2.1 mg P m-2 d-1) were 6 and 11 times
lower than the rate reported for the unamended control
soil (23 mg P m-2 d-1) in the Chimney et al. (2007)
study, indicating higher soil labile P pools may have
been present in the relict STA soil than in the WCA-
2A soil. Experimental conditions between the two
123
Wetlands Ecol Manage (2017) 25:649–666
studies also differed considerably. Our experiment
benefited from a longer monitoring period (21 months
vs. 20 days), larger platform size, flow-through con-
ditions, and intact soil cores that preserved soil
structure and vertical distribution of soil constituents.
Iron chloride
For the muck soils present at our study sites, with high
organic matter (total organic carbon: 47–48%) and
low Fe (0.11–0.12%) contents, the addition of 100 g
Fe(III) m-2 (as FeCl3) was ineffective compared to the
herbicide-only (control) treatment (Fig. 2). P export
was highest for the Fe mesocosms than any of the other
treated or Control mesocosms (Table 3). Also, there
was no significant increase (P [ 0.12; n = 2; one-
tailed) in Fe/Al-bound P at the 0-10 cm soil depths of
the Fe mesocosms from Sites N and C compared to the
initial soil analysis and the Control mesocosms
(Fig. 5). This P pool would be the one expected to
increase significantly if the amendment was binding P
released from the soil.
Our study highlights that important differences
exist between the environmental chemistry of Fe- and
Ca-based amendments that need to be taken into
consideration when selecting a chemical amendment
for soil P immobilization in wetlands. Although the
effectiveness of both amendments in controlling the
solubility of the metal-phosphate salt depends on the
pH in the environment, the ORP and sulfide concen-
tration are also critically important in the case of Fe–P
binding (Driscoll et al. 1993; Brezonik and Arnold
2011). The ORP in the Fe-treated mesocosm porewa-
ter was poised between ?6 and -152 mV, a range that
would have reduced the added Fe(III) to Fe(II) as well
as reduced sulfate to sulfide (Sherwood and Qualls
2001; Reddy and DeLaune 2008), and was indicated
by the higher dissolved Fe porewater concentrations in
the Fe(III)-amended than the Ca treatment and Control
mesocosms (Table 2). Ann et al. (2000) also reported
Fe(III) amendment to be ineffective in controlling
soluble P in muck soils of a marsh in central Florida
when soil ORP values decreased to below ?200 mV.
As in this study, they found that low ORP values
resulted in low P in the Fe/Al-bound soil pool along
with higher dissolved Fe and SRP in the porewater.
Because the mesocosms were maintained as flow-
through, and not static, systems, the added Fe(III),
which was reduced to Fe(II), was continuously being
Wetlands Ecol Manage (2017) 25:649–666
flushed from the mesocosms (Fig. A6; in Electronic
supplementary material). This depleted the Fe reser-
voir of 100 g m-2 that had initially been applied.
Some of the applied Fe also migrated to lower soil
depths as indicated by total Fe concentrations in the
10-30 cm soil depth of 2900 mg (kg dry wt)-1 (N Site)
and 3100 mg (kg dry wt)-1 (C Site) at the end of the
21 months of flow-through compared to 1250 and
1300 mg (kg dry wt)-1 prior to applying the FeCl3. As
a result, higher Fe/Al-bound P soil fractions were
observed in the 10–30 than 0–10 cm soil layers of the
Fe, relative to the Control, mesocosms (Fig. 5). Fe/Al-
bound soil P was also higher at 10–30 cm depth in the
Fe-treatment compared to the Fe/Al-bound P mea-
sured in Ca treatment or the Control.
Besides being more soluble and therefore more
likely to be flushed from the soil, reduced Fe(II) will
also preferentially bind to sulfide rather than P,
reducing its ability to immobilize P (Reddy and
DeLaune 2008). Therefore, for Fe(II) to be effective
in binding soil P, sulfate reduction and Fe(II) input
need to be in balance (Smolders et al. 1995). Although
the addition of Fe(III) decreased the porewater sulfide
concentrations compared to the Control and Ca
amendment in soils from both sites (Table 2), pore-
water sulfide concentrations were still C0.80 mg L-1
for the Fe-treated mesocosms. This excess sulfide in
the Fe-treated soils suggests the formation of insoluble
iron sulfide complexes (FeSx), accompanied by the
release of P from Fe–P complexes, leading to molar
ratios of dissolved Fe to SRP in the porewaters being
less than 3.5 for most of the Fe mesocosms (Table A2;
in Electronic supplementary material). Zak et al.
(2004, 2010) and Geurts et al. (2008) reported that
Fe:SRP molar ratios \3.0–3.5 indicate that Fe is not
contributing to SRP immobilization. Even with 100 g
Fe(III) m-2 applied, the ending soil TFe:TP molar ratio
for the 0–10 cm depth in the Fe mesocosms was less
than the 10 mol mol-1 threshold reported by Geurts
et al. (2008) as a minimum ratio for TFe to be effective
in immobilizing P. Because the soils and porewaters
from the study sites contain low native Fe concentra-
tions (1100–1200 mg (kg dry wt)-1 [0–30 cm soil
depth] and 16–90 lg L-1, respectively), while the
surface waters are relatively high in sulfate (Table 1),
the P immobilized by one-time or even periodic Fe
amendments will only be temporary since sulfide will
be continually produced in the low ORP environment.
663
Contrary to our findings, Sherwood and Qualls
(2001) were successful in inhibiting P release under
low ORP when insoluble Fe–P complexes (after
precipitating SRP with FeCl3) were applied directly
to Everglades soils in lab microcosms. They attributed
the P release inhibition to the reduction of Fe(III) to
Fe(II) to form insoluble ferrous phosphate (vivianite).
In addition to placing the pre-formed Fe–P precipitate
directly on and within the soil column, the 10-h daily
water column aeration and unknown sulfide effect
(aqueous sulfur species were not reported) likely
explain the difference in P release between our and
their study.
In addition to the low ORP and production of
sulfide, the higher pH of the pore and overlying waters
also likely contributed to the instability of Fe–P solids.
In the Fe-treated mesocosms, the standing water pH
remained around 8.0 for most of the time through the
end of the demonstration project. The porewater had a
pH 6.85–7.54, which is conducive to increasing the
solubility of FePO4 (strengite). The minimum solu-
bility of FePO4 is in the pH range of 2.5–4.5, and
solubility can increase 1000-fold from a pH of 4.5–7.0
(Brezonik and Arnold 2011).
Yet another mechanism that may have contributed
to the release of P in the FeCl3-amended mesocosms
compared to the Control mesocosms is enhanced
organic matter decomposition. Iron reduction by
facultative and obligate anaerobes is linked to organic
matter decomposition (Reddy and DeLaune 2008). By
serving as an electron acceptor in the oxidation of
organic matter, the addition of Fe(III) as FeCl3 could
have enhanced organic matter P mineralization.
Thus, a combination of low ORP, sulfide produc-
tion, accelerated organic matter turnover, and high pH
acted interactively to mitigate the P immobilization
potential of the added Fe(III) in the Fe mesocosms.
This finding was confirmed by the lower readily
available P in the 0–10 cm surface soil of the Fe-
treatments than in other chemically-treated meso-
cosms, and the lower than expected Fe/Al-bound P
pool measured in 0–10 cm soil depths from Fe-treated
mesocosms (Fig. 5). The mobilization of P from the
readily available P pool, combined with a lack of P
sequestration in the Fe/Al-bound, biogenic, and Ca/
Mg-bound pools in the Fe-amended treatment,
resulted in higher pulses of SRP, DOP, and PP from
Fe-treated soils at both sites compared to soils at either
123
664
the Control or Ca treatment mesocosms (Fig. A4; in
Electronic supplementary material).
Other than biogeochemical factors, the amendment
application methodology and operational conditions
may have also partially affected the different out-
comes between our and other studies (Smolders et al.
1995, 2001; Sherwood and Qualls 2001). Despite
capping the Fe-enriched top soil after application with
a 1-cm thick sand layer, the one-time application of
Fe(III) at 100 g m-2 was unsuccessful in immobiliz-
ing P except for a brief period in the beginning
(Fig. 2). Periodic re-application of Fe(III) may have
been more successful, but given the solubility of
Fe(III) under the reducing conditions in the soil,
washout of added Fe under flowing conditions could
still render repeated applications ineffective over the
long run.
We did not use a solid Fe chemical, which may have
been more effective since it would presumably have
been less soluble than the applied liquid Fe(III)
solution. But even in studies where solid Fe addition
prevented sulfide accumulation and resulted in SRP
immobilization (Smolders et al. 2001), the beneficial
effects were only temporary when external sulfate
loads were being received. These factors call into
question the feasibility of wide-scale application of Fe
to large areas of the P-impacted Everglades.
Conclusions
The use of intact soil-water-plant (cattail) monoliths
extracted from WCA-2A and exposed to low-P flow-
through water provided a realistic mesocosm platform
for testing the effectiveness of two soil amendments
(Fe and limerock) for minimizing soil P release to the
water column, and for evaluating the effects of a
seven-week imposed dry down followed by reflood-
ing. Intact soil columns from both locations exhibited
long-term (21 month) efflux of P when fed low P
waters. Soils from the more P-enriched Site N released
more P than soils from the less P-enriched Site C.
Higher P fluxes occurred prior to the seven-week dry
down than after reflooding during the post-dry down
period.
Our results show that Fe is not a suitable amendment
for immobilizing soil P in these marshes, since the
organic soils typically found in the impacted areas of
WCA-2A frequently have low ORPs, high pH values
123
Wetlands Ecol Manage (2017) 25:649–666
and sulfide concentrations, conditions favorable for
the reductive dissolution of ferric hydroxide with
adsorbed P and/or ferric phosphate precipitates,
resulting in the formation of FeSx and liberation of
P. The addition of Fe(III) as FeCl3 also likely served to
accelerate organic P mineralization. LR was a superior
amendment to FeCl3 in reducing P flux from soils in
WCA-2A when applied to the top of the soil column.
Even though LR is a naturally occurring substrate in
the Everglades marshes, its use in the manner
described in this study should be further evaluated
for any environmental consequences. Also, the poten-
tial costs of surficial LR application and field-scale P
immobilization benefits, would need to be more
thoroughly assessed prior to consideration of large-
scale application of this amendment.
Acknowledgements Sample analyses were performed by J.
Henry, N. Chan, K. Hileman, N. Larson, L. Colville, J. Wolack,
and S. O’Harrow. J. Potts undertook the statistical analyses.
T. Goffinet and D. Haselow helped core and transport the soil
monoliths from WCA-2A to the experimental site (SATTS).
Funding Funding was provided by the Everglades Agricul-
tural Area Environmental Protection District.
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