Spatial Gradient Analyses of Nutrients and Chlorophyll a Biomass in an

Urbanized Lagoonal Estuary: A Comparison between Wet and Dry Periods

Authors: D. L. White, D. E. Porter, A. J. Lewitus, and J. Keesee

Source: Journal of Coastal Research, 2008(243) : 649-659
Published By: Coastal Education and Research Foundation
URL: https://doi.org/10.2112/05-0498.1

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 Journal of Coastal Research 24 3 649–659 West Palm Beach, Florida May 2008

Spatial Gradient Analyses of Nutrients and Chlorophyll a

Biomass in an Urbanized Lagoonal Estuary: A Comparison
between Wet and Dry Periods
D.L. White†, D.E. Porter‡§*, A.J. Lewitus‡††, and J. Keesee‡

National Oceanographic and

†
Belle W. Baruch Institute for
‡
Department of
§
National Oceanographic and
††

Atmospheric Administration Marine and Coastal Environmental Health Atmospheric Administration

Hollings Marine Laboratory
Charleston, SC 29412, U.S.A.
whitedl@clemson.edu
Sciences
University of South Carolina
Columbia, SC 29208, U.S.A.
Sciences
Arnold School of Public
Health
University of South Carolina
Columbia, SC 29208, U.S.A.
Center for Sponsored Coastal
Ocean Research
1305 East West Highway,
Room 8220
Silver Spring, MD 20910,
U.S.A.

ABSTRACT
WHITE, D.L.; PORTER, D.E.; LEWITUS, A.J., and KEESEE, J., 2008. Spatial gradient analyses of nutrients and
chlorophyll a biomass in an urbanized lagoonal estuary: a comparison between wet and dry periods. Journal of Coastal
Research, 24(3), 649–659. West Palm Beach (Florida), ISSN 0749-0208.

The effects of stochastic events on estuarine water quality parameters are often difficult to quantify spatially due to
the inherent variability in these systems. The use of geographic information systems to identify spatial patterns and
trends can improve such efforts. This work describes the use of geographic information systems and the Mantel Test
to determine the existence of trends and the persistence of spatial patterns in the relationships between nutrients
and chlorophyll a in an urbanized lagoonal estuarine system, Murrells Inlet, South Carolina. Relatively dry and wet
periods were compared, the latter following several substantial precipitation events related to tropical hurricane
activity in the summer of 1999. Dissolved inorganic nitrogen concentrations were elevated following the passage of
Hurricane Irene, and chlorophyll a did not increase concomitantly. Orthophosphate was found to have persistently
higher concentrations in the more urbanized regions of Murrells Inlet that were consistent spatially during the two
sampling periods. Strong spatial and temporal correlations between chlorophyll a and dissolved silicate : dissolved
inorganic nitrogen suggest a relationship between the latter ratio and the phytoplankton growth. However, there
appeared to be only a tenuous relationship between chlorophyll a and dissolved inorganic nitrogen and phosphorus,
both spatially and temporally, suggesting that nutrient requirements for phytoplankton growth were met by internal
estuarine processes, during this time period.

ADDITIONAL INDEX WORDS: Chlorophyll, geographic information systems (GISs), nutrients, hurricane, water qual-
ity, spatial analysis.

INTRODUCTION The effects of urbanization on estuarine eutrophication are

The identification and analysis of spatial clusters and
trends in natural systems can provide insight into ecological
phenomena because such patterns may be structured by com-
mon environmental processes (LENGENDRE and LENGENDRE,
1998). The presence of a spatial gradient may reflect the ef-
fects of broad-scale physical processes or ‘‘forcing functions’’
that create gradients in the physical environment (LENGEN-
DRE and TROUSSELLIER, 1988). Identification of common spa-
tial patterns or correlations among environmental variables
may indicate a common spatial gradient. In some situations,
these gradients may be indicated by a trend such as a de-
creasing range of values in a geographic direction or other
spatial patterns that show some consistency over time and
may indicate larger ecological phenomena.
DOI:10.2112/05-0498.1 received 27 April 2005; accepted in revision
17 November 2006.
* Present address: Clemson University, ITC, 340 Computer Court,
Anderson, SC 29625, U.S.A.
complex and difficult to assess. However, the detection and
analysis of spatial patterns may provide insight into the re-
lationships between urbanization-associated landscape
changes and estuarine temporal and spatial variability. To
better understand the dynamics of spatial gradients in es-
tuaries and how they might be affected by stochastic events,
a high-salinity estuarine system (Murrells Inlet, South Car-
olina) was sampled during a relatively dry period in the sum-
mer of 1999 and again in the fall of 1999 following the nearby
passage of three hurricanes: Hurricane Dennis (24 August–5
September), Hurricane Floyd (14–17 September), and Hur-
ricane Irene (13–16 October). Significant portions of North
Carolina and the northeastern coastal counties of South Car-
olina (SC) were particularly impacted by significant flooding
events during this latter time period (BALES, 2003; BURK-
HOLDER et al., 2004; MALLIN et al., 2002; PEIERLS, CHRIS-
TIAN, and PAERL, 2003; RAMUS et al., 2003).
Murrells Inlet (MI) is a high-salinity lagoonal estuary rep-
resentative of similar coastal areas that are increasingly in-
fluenced by anthropogenic development. In SC, small high-

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 650 White et al.

salinity creeks and inlets exceed the total combined area of

larger river-dominated systems (VERNBERG et al., 1992).
Shallow lagoonal-type environments that lack significant
freshwater inputs generally have lower nutrient levels than
riverine-dominated estuaries (NIXON, 1982). Phytoplankton
and nutrient concentrations on a systemwide scale have been
assumed homogeneous in these estuaries, based on the char-
acteristically high mixing and flushing properties. However,
empirical data sets to support this supposition are lacking,
because spatially extensive temporal studies are rare in these
systems.
In this study, spatially interpolated surfaces that charac-
terize water quality variables in MI—nitrate (NO⫺ 3 ), ortho-
phosphate (PO⫺ ⫹
4 ), ammonium (NH4 ), chlorophyll a (Chl a),
and silicate : dissolved inorganic nitrogen (Si : DIN)—are de-
veloped and discussed. These data are spatially intensive and
were collected during relatively dry and wet periods. These
two distinct periods provide an opportunity to investigate
spatial relationships under differing environmental condi-
tions. We hypothesized that (1) water quality variables are
strongly influenced by the diverse landscape in MI, leading
to gradients or clustering of these parameters; (2) Chl a is
spatially correlated to inorganic nutrients; and (3) spatial
gradients will intensify during the wet period as urban runoff
leads to increased nutrient inputs.

MATERIALS AND METHODS

Study Site and Methods
MI is a tidally dominated, bar-built estuary characterized
by extensive Spartina alterniflora habitat and high salinities.
The estuary encompasses approximately 3000 acres of
marshland, and the upland areas have been developed to ac-
commodate an increasing flux of residents and tourists
(VERNBERG et al., 1992). The estuary is surrounded by a mix
of residential and commercial development on all sides with
the exception of Huntington Beach State Park to the south-
east (Figure 1). The estuary has one exchange point with oce-
anic waters and has nine outfall pipes draining the upland
watersheds. MI has an average depth of 1.5 m and a maxi-
mum depth of 10.0 m during midtide (B. JONES and D. POR-
TER, unpublished data).
Thirty sites were sampled in MI (Figure 1) on 24 August
1999 (AUG) and 22 October 1999 (OCT). Data collections
were carried out by two teams, one covering the northern
extent and the other covering the southern extent. Water
samples were collected approximately 0.5 m below the sur-
face using grab samples in acid-washed, polyethylene, 1-L
bottles. Three replicate bottles were collected at each sample
location. Inorganic nutrients (NO⫺3 ⫹ NO2 ⫽ NO3 , NH4 , SiO4
⫺ ⫺ ⫹
⫽ Si, PO4 ) were determined by automated colorimetric anal-
⫺3
yses using Technicon AutoAnalyzers. Chlorophyll a was mea-
sured fluorometrically after extraction in 90% acetone by a
freeze–thaw method (LEWITUS, KOEPFLER, and MORRIS,
1998). Daily (24 h) precipitation data were obtained from the
National Weather Service for MI, and daily totals were
summed to determine monthly totals. Salinity was measured
using a refractometer, and the mean among sampling sites
was calculated.
Figure 1. Murrells Inlet land use/cover and collection sites for the 24
August 1999 and 22 October 1999 data collections.
Spatial Interpolation
The analysis of spatial structures was initiated by spatial
interpolation of NO⫺ ⫺3 ⫹
3 , PO4 , NH4 , Chl a, and Si : DIN for the
AUG and OCT data, using radial basis functions (RBFs).
RBFs form a spatial interpolation technique available in the
ESRI ArcMap娂 software. The RBF interpolators are similar
to neural networks and allow the user some control over the
parameter settings. RBFs are moderately deterministic in-
terpolators that are exact, meaning that the predicted surface
must go through the sample locations. However, RBFs do not
account for spatial autocorrelation, thus making them less
flexible than some other interpolators, such as Kriging, that
will account for an autocorrelation structure and produce er-
ror or uncertainty surfaces (ESRI, 2001). The RBF is using
Euclidean distance between data points instead of in-creek
distance for calculating the predicted surface. Previous re-
search in MI compared the use of Euclidean distances vs. in-
creek distances and found improvement in prediction accu-
racy for some environmental parameters under study; how-
ever, overall, the use of in-creek distances resulted in unpre-

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 Spatial Gradients 651

dictable and modest improvements (LITTLE, EDWARDS, and

PORTER, 1997). In addition, spatial autocorrelation was ex-
amined using semivariogram analyses for all of the parame-
ters under study, and in many instances the semivariograms
were found to be inconclusive (D. WHITE, unpublished data).
Thus, Kriging was not a good choice as an interpolation tech-
nique. To be consistent, the RBF was viewed as a means to
apply a consistent interpolation technique. RBFs make no as-
sumptions about the data and therefore are an adequate
method to develop initial interpolated surfaces. A weakness
of RBFs is that they are primarily suited for moderate chang-
es in surface values; therefore, large changes in short dis-
tances may invalidate this technique. A built-in cross-vali-
dation technique was used to calculate a root-mean-square
error (RMSE) for each interpolated model.
The interpolation methods were initiated by calculating
means for each parameter at each sample location. These
data were combined with latitude and longitude coordinates
and subsequently imported into ArcGIS as comma-separated
value files using the ArcCatalog functions. The projections for
the data were set to a Universal Transverse Mercator NAD83
coordinate system. Using the Geostatistical Wizard extension
in ArcMAP, the data were interpolated using RBF and a reg-
ularized spline function with a nearest neighborhood search
of 15 points for each variable. An anisotropic search area was
developed to account for the geographic orientation of MI that
roughly follows a 45⬚ trend (Figure 1). After interpolation, the
continuous surface was converted to a grid and followed by
raster calculations to allow for a conversion to an ESRI
shapefile. The grid was a floating point file that precludes
the conversion to an ESRI coverage file. To work around this,
the grid values were multiplied by 1000 and then converted
to integers. This grid file was converted first to an ESRI cov-
Figure 2. Creek network and sampling locations used for the GIS-based
erage file and then to an ESRI shapefile. This output file was
network analyses.
rectangular and covered a much greater area than that of the
area studied (i.e., the MI creek system). A data layer of the
creek system was therefore used to extract a data layer that
was specific to the creek system. The models are presented in a geographic information system (GIS). Digitizing the MI
in a manner to highlight spatial structures across the two creek system in the ESRI ArcView 3.2娂, a vector data layer
sampling dates, so in some cases, the classification range may of the creek system was created focusing on the primary and
not be ecologically significant. secondary creeks where the collection sites are located. The
source data originated from a detailed creek boundary file
Mantel Tests originally created by digitizing 1 : 4800-scale CIR photos ac-
quired at low tide during the early (leaf off) part of 1988 (D.
Interpolated surfaces are useful for identifying trends and PORTER, personal communication). The data locations deter-
patches in a spatial context. However, these types of analyses mined via a global positioning system (GPS) survey of the
are limited with regard to verifying or interpreting the pro- collection sites were also entered into this GIS (Figure 2). In
cesses affecting visible spatial patterns that appear within this context, primary creeks were navigable at low tide while
and between each collection period. The Mantel Test (MAN- secondary creeks branched from primary creeks and may or
TEL, 1967) was used to better understand the spatial and may not be navigable at low tide. These two data layers fa-
temporal patterns observed through the interpolated surfac- cilitated a network analysis using the Arc Script Shortest
es. This nonparametric analysis tests the null hypothesis that Network Paths v1.1 (REMINGTON, 1999) that calculated the
two independent similarity, dissimilarity, or distance matri- nearest creek distance between collection sites that were sub-
ces are not significantly different (L ENGENDRE and LENGEN- sequently exported from GIS into a matrix table (DIS). The
DRE, 1998; SOKAL and ROHLF, 1995). Two types of matrices second set of matrices consists of dissimilarity matrices cal-
were developed, including a matrix composed of the geo- culated by taking the unsigned difference among the mean
graphic distances between all of the collection sites based values of the parameters (VAR) of interest (NO⫺ ⫹
3 , NH4 ,
upon creek distances between the 30 collection sites. These PO⫺34 , Chl a, and Si : DIN), following the methods described
distances were generated from a network analysis performed by LENGENDRE and TROUSSELLIER (1988).

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 652 White et al.

These matrices were used to test both spatial and temporal

hypotheses. The first set of spatial hypotheses that were ex-
amined used the DIS matrix and the dissimilarity matrices
(VAR) of each parameter examined. The purpose of this anal-
ysis is to determine whether a linear relationship exists be-
tween parameter concentrations and their spatial location.
Thus, the null hypothesis for these tests is that there are no
significant spatial trends in the distribution of the parame-
ters under investigation. The spatial distributions of the pa-
rameters were also examined for spatial correlation between
the VAR matrices within and between collection periods, thus
exploring the spatial correlations between the parameters
over time and space. The null hypothesis for these analyses
is that there is no spatial correlation between parameters
within and between collection periods.
The basic form of the Mantel statistic, zm, is calculated from
the sum of cross products of the (unstandardized) values in
the two matrices excluding the main diagonal (which were
zeros because distance and dissimilarity matrices were cal-
culated; LENGENDRE and LENGENDRE [1998]. If dissimilari-
ties or distances correspond in magnitude between the two
data matrices, then zm will be large and indicate a positive
association. If the magnitudes of these values are dissimilar
from one another, then the correlation is expected to be small
(SOKAL and ROHLF, 1995). The current study also presents
a correlation coefficient (r) and the g statistic (standard nor-
mal variate). These are one-tailed tests because the hypoth-
esis tests the positive relationship between the two variables;
thus, the levels of significance that have been calculated for
the g statistic are based on the upper tail.
The Mantel statistics are tested by permutation (also
known as randomization), which is a general approach of
testing statistical hypotheses without needing to meet the
distributional assumptions of any particular statistical meth- Figure 3. Monthly mean precipitation and salinity from Murrells Inlet
(a) precipitation and (b) salinity. Error bars are ⫾1 SE (salinity only).
od, such as a normal distribution (LENGENDRE and LENGEN- Sampling periods in the present study indicated by an open circle (䡬).
DRE, 1998). In permutation, the reference distribution is gen-
erated from a randomization of the original data. The refer-
ence distribution is a composite of these randomizations to
generate a distribution that compares to the original data. were made after a relatively dry period, while the OCT col-
Thus, it serves a role similar to the normal distribution in lections occurred after significant rain events in September
parametric statistical analyses. The test statistic is compared and October during the passage of Hurricane Floyd and Hur-
to this reference distribution. There are two steps to the per- ricane Irene. The highest recorded precipitation levels were
mutation analysis, with the initial recognition that the ob- in September 1999 during the passage of Hurricane Floyd (15
served sample of parameters is just one of many possible, but September 1999). From 16–21 October, there was approxi-
equally likely, different outcomes that could have arisen due mately 216 mm of precipitation, primarily the result of the
to chance. These other outcomes are then enumerated by ran- passage of Hurricane Irene over the SC coast approximately
domly rearranging the parameters, and, on the basis of the 3 to 5 days prior to the OCT data collections. These rainfall
resulting distribution, the observed outcome is tested to de- events are reflected in marked salinity differences between
termine whether it is dissimilar enough to warrant rejection the AUG and the OCT sampling periods (Figure 3b).
of the null hypothesis (SOKAL and ROHLF, 1995). Thus, 1000 These stochastic events had significant impacts on nutrient
random iterations were generated out of a possible 1 million inputs into the estuarine systems, increasing the concentra-
with an alpha level of 0.05. Mantel Tests were performed tions of all inorganic nutrients (Figures 4–6; see also WHITE,
using the shareware Mantel v2.0 (LIEDLOFF, 1999). PORTER, and LEWITUS, 2004). A spatial gradient was appar-
ent, indicating higher nutrient concentrations in the northern
RESULTS regions of the estuary. Lower concentrations were associated
with the inlet and southern reach of MI. The PO⫺3 4 data ap-
Interpolated Surfaces (Maps)
peared to be the most consistent in regard to the spatial pat-
Rainfall patterns prior to the sampling dates differed be- terns over time (Figure 6). Although concentrations were
tween collection periods (Figure 3a). The AUG collections higher during the OCT sampling period, both interpolated

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 Spatial Gradients 653

Figure 4. Interpolated surfaces of NO⫺3 for the Murrells Inlet creek sys-
tem: (a) collections from 24 August 1999 and (b) collections from 22 Oc-
tober 1999. Legends are meant to assist in the identification of trends;
thus, color scheme and class breakdown are not presented in a continuous
manner between collection dates. For a color version of this figure, see
page 690.
Figure 5. Interpolated surfaces of NH⫹4 for the Murrells Inlet creek sys-
tem: (a) collections from 24 August 1999 and (b) collections from 22 Oc-
tober 1999. Legends are meant to assist in the identification of trends;
thus, color scheme and class breakdown are not presented in a continuous
manner between collection dates. For a color version of this figure, see
page 690.

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 654
Figure 6. Interpolated surfaces of PO⫺3
4 for the Murrells Inlet creek sys-
tem: (a) collections from 24 August 1999 and (b) collections from 22 Oc-
tober 1999. Legends are meant to assist in the identification of trends;
thus, color scheme and class breakdown are not presented in a continuous
manner between collection dates. For a color version of this figure, see
page 691.
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White et al.
surfaces indicated ‘‘hotspots’’ in the same general locations
during both sampling periods. These areas are located in the
urbanized region.
The Chl a data (Figure 7) show some similarities over time,
with higher concentrations found in the smaller creeks along
the southwestern region of the estuary. The reported Chl a
concentrations for AUG corresponded to annual maxima re-
ported by WHITE, PORTER, and LEWITUS (2004). Higher con-
centrations were apparent in the more urbanized region dur-
ing the AUG collection period. During OCT, a similar trend
was observed, with the highest concentrations observed in
the southwest region of MI. The Si : DIN data appeared to
track Chl a, with the greatest ratio of Si : DIN occurring in
the southwest and north–northwest regions of MI during the
AUG collection period (Figure 8a). The spatial gradients of
Si : DIN were also similar to the Chl a data during OCT, with
the highest magnitudes observed in the south–southwest re-
gion (Figure 8b). Overall, these data suggest that the north-
ern regions of MI had higher inorganic nutrient concentra-
tions. However, the Chl a and Si : DIN data did not follow the
inorganic nutrient trends because highest concentrations
generally occurred along the south–southwestern boundary
of the estuary.
The cross-validation procedure in ArcMap娂 uses all of the
data to estimate a trend model. A data point is removed, one
at a time, and a new predicted value is calculated using the
remaining values. This is repeated for all of the data points,
and the measured and predicted values are compared. The
overall performance of the model can be examined by looking
at the RMSE; as a value increases from zero, that is an in-
dication of increasing error in model performance. The units
of RMSE are the same as the parameter of interest, thus
comparing RMSE is only useful with models that are of the
same units. For the AUG sampling period, RMSE values for
NO⫺ 3 , NH4 , PO4 , Chl a, and Si : DIN were 0.13 ␮M, 1.58 ␮M,
⫹ ⫺3
0.08 ␮M, 3.59 ␮g/L, and 3.63, respectively. RMSE values for
the OCT analyses of NO⫺ ⫹ ⫺3
3 , NH4 , PO4 , Chl a, and Si : DIN
were 0.69 ␮M, 2.39 ␮M, 0.06 ␮M, 1.04 ␮g/L, and 0.42, re-
spectively. Overall, model performance for NO⫺ 3 and NH4
⫹
suggests that as concentrations increased during OCT model
performance decreased. However, PO⫺3 4 was relatively stable
between AUG and OCT and showed the best performance of
all models. Chlorophyll a decreased between AUG and OCT
as concentrations decreased; this trend was also observed for
Si : DIN.
Mantel Tests
The Mantel analyses were designed to test for similarities
and trends between the parameters in a spatiotemporal con-
text. A particularly well-developed spatial gradient for the
inorganic nutrients was apparent, with higher concentrations
observed in the northern regions of the estuary followed by
reduced concentrations toward the southern extent and the
inlet region. To test the null hypotheses that there were not
linear trends for each of the parameters examined, Mantel
Tests were computed (VAR ⫻ DIS). Mantel results indicated
the presence of a significant linear trend only for PO⫺3
4 during
the AUG collection period (Table 1). Significant linear trends
 Spatial Gradients 655

Figure 7. Interpolated surfaces of Chl a for the Murrells Inlet creek
system: (a) collections from 24 August 1999 and (b) collections from 22
October 1999. Legends are meant to assist in the identification of trends;
thus, color scheme and class breakdown are not presented in a continuous
manner between collection dates. For a color version of this figure, see
page 691.
Figure 8. Interpolated surfaces of Si : DIN for the Murrells Inlet creek
system: (a) collections from 24 August 1999 and (b) collections from 22
October 1999. Legends are meant to assist in the identification of trends;
thus, color scheme and class breakdown are not presented in a continuous
manner between collection dates. For a color version of this figure, see
page 692.

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 656 White et al.

Table 1. Mantel statistics for parameter comparisons during the August 24, 1999 collection event. Probabilities are for the g statistic (standard normal
variate) and are one-tailed based on an alpha of 0.05.

NO3 (VAR) NH4 (VAR) PO4 (VAR) Chl a (VAR) Si:DIN (VAR) DIS

NO3 (VAR) — r ⫽ 0.093 r ⫽ 0.272 r ⫽ ⫺0.061 r ⫽ ⫺0.053 r ⫽ 0.106

g ⫽ 0.662 g ⫽ 2.148 g ⫽ ⫺0.477 g ⫽ ⫺0.393 g ⫽ 1.314
P ⫽ 0.254 P ⫽ 0.0159 P ⫽ 0.6833 P ⫽ 0.6528 P ⫽ 0.094
NH4 (VAR) — — r ⫽ 0.151 r ⫽ 0.106 r ⫽ 0.061 r ⫽ 0.095
g ⫽ 1.289 g ⫽ 0.888 g ⫽ 0.491 g ⫽ 1.252
P ⫽ 0.0989 P ⫽ 0.1873 P ⫽ 0.3117 P ⫽ 0.1503
PO4 (VAR) — — — r ⫽ ⫺0.031 r ⫽ 0.016 r ⫽ 0.273
g ⫽ ⫺0.282 g ⫽ 0.146 g ⫽ 3.836
P ⫽ 0.611 P ⫽ 0.442 P ⫽ 0.0001
Chl a (VAR) — — — — r ⫽ 0.645 r ⫽ 0.099
g ⫽ 5.650 g ⫽ 1.372
P ⬎ 0.0001 P ⫽ 0.085
Si:DIN (VAR) — — — — — r ⫽ 0.051
g ⫽ 0.692
P ⫽ 0.2445
r ⫽ correlation between any two parameters, g ⫽ standard normal variate.

were identified for all of the parameters during OCT (Table
2).
There is interest not only in identifying spatial trends but
also in identifying relationships among the parameters dur-
ing the sampling periods, which may indicate physical or bi-
ological relationships. Mantel Tests were used to test the null
hypotheses that there were not similar spatial patterns be-
tween the parameters of interest. The VAR ⫻ VAR analyses
indicated two significant relationships for the AUG matrices,
NO⫺ 3 ⫻ PO4 and Chl a ⫻ Si : DIN (Table 1). The data from
⫺3
the OCT analyses indicated more spatial interaction between
the parameters. Nitrate and NH⫹ 4 were significantly related
to PO⫺34 , but NO3 ⫻ NH4 was not statistically significant
⫺ ⫹
(Table 2). Nitrate was significantly related to Chl a and Si :
DIN, and Chl a ⫻ Si : DIN was significantly related.
Further analyses testing the null hypothesis that there is
no association or correlation between the parameter matrices
over time found that the PO⫺3 4 matrices were significantly
similar between the AUG and the OCT collection periods (Ta-
ble 3). These results support the observations of PO⫺3 4 ‘‘hot-
spots’’ in the northern region of the MI. In addition, the Chl
a and Si : DIN matrices were found to be significantly related
over time.
DISCUSSION
Overall, the nitrogen (N) and phosphorus (P) spatial trends
are likely a function of nutrient inputs from urban runoff,
groundwater, and dilution from coastal waters. This is sup-
ported by previous research in MI that looked at the potential
spatial contributions of upland runoff and coastal waters in
affecting water quality and Chl a (WHITE, PORTER, and LEW-
ITUS, 2004). The surrounding landscape of MI is highly var-
iable, with more developed upland areas surrounding the
northern regions of the estuary and a state park to the south.
The inlet is a source for coastal waters that are nutrient poor
relative to the estuarine water. Mantel Test results indicated
intensification of the gradients for N and P during OCT, fol-
lowing the heavy rainfall in late September and October. Be-
cause of the lack of significant riverine freshwater sources in
the MI estuary, nonpoint source pollution runoff is likely the
main source of ‘‘new’’ inorganic N and P following rain events.

Table 2. Mantel statistics for parameter comparisons during the October 22, 1999 collection event. Probabilities are for the g statistic (standard normal
variate) and are one-tailed based on an alpha of 0.05.

NO3 (VAR) NH4 (VAR) PO4 (VAR) Chl a (VAR) Si:DIN (VAR) DIS

NO3 (VAR) — r ⫽ ⫺0.022 r ⫽ 0.192 r ⫽ 0.576 r ⫽ 0.500 r ⫽ 0.207

g ⫽ ⫺0.178 g ⫽ 2.061 g ⫽ 4.615 g ⫽ 4.407 g ⫽ 2.883
P ⫽ 0.5706 P ⫽ 0.0197 P ⬎ 0.0001 P ⬎ 0.0001 P ⫽ 0.002
NH4 (VAR) — — r ⫽ 0.614 r ⫽ ⫺0.139 r ⫽ ⫺0.082 r ⫽ 0.296
g ⫽ 5.814 g ⫽ ⫺0.966 g ⫽ ⫺0.630 g ⫽ 3.731
P ⬎ 0.0001 P ⫽ 0.833 P ⫽ 0.7357 P ⫽ 0.0001
PO4 (VAR) — — — r ⫽ 0.018 r ⫽ ⫺0.018 r ⫽ 0.370
g ⫽ 0.170 g ⫽ ⫺0.185 g ⫽ 5.724
P ⫽ 0.4325 P ⫽ 0.5734 P ⬎ 0.0001
Chl a (VAR) — — — — r ⫽ 0.720 r ⫽ 0.158
g ⫽ 5.558 g ⫽ 1.998
P ⬎ 0.0001 P ⫽ 0.0229
Si:DIN (VAR) — — — — — r ⫽ 0.175
g ⫽ 2.373
P ⫽ 0.009
r ⫽ correlation between any two parameters, g ⫽ standard normal variate.

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 Spatial Gradients
Table 3. Mantel statistics for parameter comparisons across the August 24, 1999 and October 22, 1999 collection events. Probabilities are for the g statistic
(standard normal variate) and are one-tailed based on an alpha of 0.05.
NO3 (VAR) NH4 (VAR)
NO3 (VAR) r ⫽ 0.007 — —
g ⫽ 0.051
P ⫽ 0.4797
NH4 (VAR) — r ⫽ ⫺0.012
g ⫽ ⫺0.090
P ⫽ 0.5359
PO4 (VAR) — —
Chl a (VAR) — —
Si:DIN (VAR) — —
r ⫽ correlation between any two parameters, g ⫽ standard normal variate.
Nonpoint sources generally include fertilizers, animal
waste, sediments, storm runoff, groundwater, and atmo-
spheric deposition (JAWORSKI, HOWARTH, and HETLING,
1997; JORDAN, CORRELL, and WELLER, 1997; PAERL, 1997).
These nutrient sources may be funneled into the estuary by
impervious surfaces such as roads, driveways, parking lots,
rooftops, and other common landscape features of an urban
environment (U.S. ENVIRONMENTAL PROTECTION AGENCY
STAFF, 1997; WASCHBUSCH, SELBIG, and BANNERMAN,
2000). The diverse landscape features that surround MI im-
pose different conditions on the estuary, resulting in regions
that receive varying degrees of nutrient runoff. Investigators
have shown that these systems are susceptible to modest
changes in the upland cover, with measurable adverse chang-
es in the physical and chemical environment observed at 10
to 20% impervious surface within a watershed (HOLLAND et
al., 2004). Analyses have shown a decreasing gradient in im-
pervious surface from the northern region to the southern
region in MI (TUFFORD et al., 2003; D. WHITE, unpublished
data). Higher concentrations found in the northern regions
are associated with the urbanized features. Concentrations
decrease in the southern region, and a relatively nutrient-
poor region is associated with the inlet. Thus, three regions
in MI can be differentiated based on coastal and upland pro-
cesses, including (1) a northern region defined by higher nu-
trient concentrations; (2) a southern region with moderate
nutrient concentrations; and (3) the inlet region character-
ized by relatively low nutrient concentrations.
The interpolated maps indicate hotspots and clusters of nu-
trient concentrations that appear to be stable over time. This
was particularly apparent for PO⫺3 4 , although further long-
term studies are needed to confirm this. Previous research
has shown fecal coliforms to be elevated in specific regions in
MI within the general proximity of the observed hotspots for
this study (KELSEY et al., 2004). As noted previously, the
northern region of MI is characterized by various urbanized
features that may be affecting specific locations within the
estuary. In addition, the freshwater streams and ponds that
drain into MI were found to have higher NO⫺ 3 and total P
concentrations compared with a reduction in organic N for
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657
PO4 (VAR) Chl a (VAR) Si:DIN (VAR)
— —
— — —
r ⫽ 0.599 — —
g ⫽ 6.509
P ⬎ 0.0001
— r ⫽ 0.528 —
g ⫽ 4.231
P ⬎ 0.0001
— — r ⫽ 0.531
g ⫽ 4.498
P ⬎ 0.0001
these particular areas due to development when compared to
pristine forested watersheds of a nearby estuarine system
(TUFFORD et al., 2003). The Mantel Tests indicated a signif-
icant correlation between the AUG and the OCT data. This
stability is surprising since the AUG data were collected dur-
ing a relatively dry period and the OCT data were collected
immediately after a period of heavy rainfall, resulting in the
highest N concentrations observed in MI during the course of
an approximate 2-year study (WHITE, PORTER, and LEWITUS,
2004). Orthophosphate concentrations were generally less
than 1 ␮M during the dry and wet periods. Orthophosphate
has virtually no atmospheric phases and stays largely in the
water column and in marsh sediments composed of clay sed-
iments and peat (FROELICH, 1988; WHITNEY et al., 1981).
These physical processes may have implications in the ap-
parent lack of spatial variability for PO⫺3
4 in MI.
The interpolated maps for Chl a and Si : DIN were very
similar during AUG and OCT, suggesting a strong spatiotem-
poral relationship. Previous research in MI has shown posi-
tive temporal correlations between Chl a and silicate (WHITE,
PORTER, and LEWITUS, 2004). The strong correlations ob-
served between Chl a and Si : DIN were likely a reflection of
diatom growth. Silicate is not limiting in these systems, and
N was not likely to be limiting during the summer, when N
concentrations reach annual maxima (LEWITUS, KOEPFLER,
and MORRIS, 1998; WHITE, PORTER, and LEWITUS, 2004).
Therefore, the Si : DIN pattern may be a reflection of phyto-
plankton uptake and remineralization of silicate and N in the
sediments and water column. Phytoplankton assemblage
data indicate that diatoms are dominant in this system and
the spatial distributions of diatoms tend to follow those of
Chl a (LEWITUS et al., 2005; D. WHITE, unpublished data).
The lowest Chl a concentrations were associated with the
inlet region of MI, consistent with general findings that
southeastern U.S. coastal waters have lower phytoplankton
biomass than adjacent estuaries. Maximum Chl a concentra-
tion was consistently centered within a creek located on the
southwestern fringe of the estuary, although peaks in the
more northern areas of MI occurred during AUG. Spatial sta-
bility was observed for Chl a and Si : DIN despite the strong
 658
tidal flushing characterizing this system. However, this par-
ticular creek does not appear to be natural and is bordered
by several houses with small docks that support water access
for residents. Observations in this area suggest this partic-
ular section may not flush as well as the primary creeks in
MI. In addition, based on documented linkages between
coastal development and eutrophication (KENNISH, 1992;
PINCKNEY et al., 2001), spatial correlation between elevated
nutrient concentrations and phytoplankton growth may be
expected, assuming upland nutrient inputs were a significant
and critical source for new N and P.
As was common with the inorganic nutrients for the OCT
period, the VAR ⫻ DIS comparisons were significant, indi-
cating linear trends for Chl a and Si : DIN. However, the in-
terpolated maps indicated that the trend was reversed for
Chl a and Si : DIN. Also, the Mantel results indicated that
Chl a and Si : DIN were not significantly correlated to NH ⫹ 4
or PO⫺34 (either positively or negatively) during these two pe-
riods. Nitrate was significantly correlated with Chl a and Si :
DIN during OCT. The interpolated NO⫺ 3 data for OCT sug-
gest that the strong clustering observed previously for AUG
decreased to some extent during OCT. The spatial correla-
tions between Chl a and NO⫺
3 may be not a result of a bio-
logical relationship but a function of increased NO⫺
3 inputs
near the marsh–upland interface where biomass was elevat-
ed. The apparent disconnect between upland nutrient inputs
and phytoplankton biomass may be due to internal estuarine
processes meeting phytoplankton nutritional requirements,
primarily remineralization. LEWITUS, KOEPFLER, and MOR-
RIS (1998) demonstrated the importance of regenerated nu-
trients to phytoplankton in summer in a similar estuary,
North Inlet, SC. NOBLE et al. (2003) also presented evidence
that the dominance of autochthonous factors (e.g., regener-
ated nutrients and grazing) over allochthonous factors (e.g.,
nutrient loading and changes in turbidity) in regulating phy-
toplankton biomass in North Inlet in the summer was related
to a relatively small watershed and lack of riverine fresh-
water input compared to another minimally urbanized salt
marsh estuary in South Carolina, the Ashepoo–Combahee–
Edisto Basin. Nitrogen loading would be expected to impact
phytoplankton nutrition to a greater degree in the winter and
spring, when population growth is N limited in North Inlet
and therefore presumably in MI. Furthermore, the location
and stability of the Chl a maxima in our study may be related
to physical properties. The spatial interpolation maps showed
higher biomass generally associated with the smaller creeks
near the marsh–upland interface. These creeks appeared to
be somewhat isolated from the primary tidal creeks in MI,
which may affect tidal flow, minimizing tidal flushing and
resulting in decreased losses of localized phytoplankton bio-
mass.
CONCLUSIONS
The use of the interpolation maps highlighted water qual-
ity patterns in the MI system, and this is the first step in the
identification of trends and patterns. These maps, combined
with the Mantel Test, allowed the testing of spatial and tem-
poral hypotheses that facilitated the diagnosis and interpre-
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White et al.
tation of estuarine processes. These methods also provided
the opportunity to formulate new hypotheses that may have
implications outside of this system. Of importance is the ex-
amination of the possible internal processes that may be af-
fecting the stability of the Chl a and PO⫺34 concentrations in
spatiotemporal context. The lack of an observed increase in
phytoplankton biomass concomitant with increased nutrient
loading during the summer may reflect a dominant influence
of autochthonous regulation of phytoplankton growth (e.g.,
nutrient regeneration), but potential other influences include
rapid recovery related to high tidal flushing rates or light
limitation of phytoplankton growth resulting from increased
turbidity (BURKHOLDER et al., 2004; MALLIN et al., 1999,
2002; NOBLE et al., 2003). Application of these analyses to
winter–spring periods is needed to test whether the influence
of stochastic events on the distribution and magnitude of phy-
toplankton biomass depends on their seasonal timing.
ACKNOWLEDGMENTS
The research described within was supported in part by a
Graduate Research Fellowship from National Estuarine Re-
search Reserve System (grant NA07OR0725), the Slocum-
Lunz Foundation (proposal R-98㛮115600), the South Carolina
Space Grant Consortium, and National Oceanic and Atmo-
spheric Administration grant NA90AA-D-SG672. In addition,
we wish to acknowledge the efforts of students and staff as-
sociated with the Baruch Institute Geographic Information
Processing Laboratory at the University of South Carolina
who assisted in data collection and Bill Johnson of the Ba-
ruch Marine Field Laboratory and Dr. Don Edwards of the
University of South Carolina for their efforts and guidance.
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