Marine Pollution Bulletin 192 (2023) 115007

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Marine Pollution Bulletin

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

Heavy metal compartmentalisation in salt marsh and seagrass of the

urbanised Swartkops estuary, South Africa
Marelé A. Nel a, b, c, Janine B. Adams a, c, Gletwyn Rubidge d, Lucienne R.D. Human b, c, *
a
DSI/NRF Research Chair: Shallow Water Ecosystems, Nelson Mandela University, Summerstrand South Campus, Gqeberha, South Africa
b
South African Environmental Observation Network (SAEON) Elwandle Coastal Node Nelson Mandela University, Ocean Sciences Campus, 4 Gomery Avenue,
Summerstrand, Gqeberha 6031, South Africa
c
Department of Botany, Nelson Mandela University, Summerstrand, South Campus, PO Box 77 000, Gqeberha 6031, South Africa
d
Department of Chemistry, Nelson Mandela University, Summerstrand North Campus, PO Box 77 000, Gqeberha 6031, South Africa

A R T I C L E I N F O A B S T R A C T

Keywords: Wetland plants are naturally exposed to high metal concentrations, and often have mechanisms to prevent metal
Wetland toxicity. This study compared metal concentrations in seagrass (Zostera capensis) and salt marsh (Spartina mar­
TXRF itima, Salicornia tegetaria) — to determine their niche as metal sinks. Samples were collected in each season over a
Phytoremediation
year at five sites in the estuary and analysed using a Total X-Ray Fluorescence (TXRF) spectrometer. Spartina
Heavy metals
maritima and S. tegetaria accumulated in their roots, and displayed little translocation to leaves (BCF = 1–14;
Sink
TF[leaf/root] < 1). Contrastingly, Z. capensis showed high uptake to its leaves (TF[leaf/root] = 1–8; Mn, Zn, Cr, Pb, Ni,
Cu). Spartina maritima and S. tegetaria were identified as good phytostabilisers (BCF > 1, TF[leaf/root] < 1).
Compartmentalization was unique to each species, and considering their ecosystem service importance, more
plant species should be analysed to estimate their ecological value for management purposes.

1. Introduction
Higher degree of pollution input is globally recorded in marine and
estuarine ecosystems, including trace elements such as heavy metals
(Sánchez-Quiles et al., 2017; Li et al., 2022). Unlike other organic pol­
lutants, metals persist in the environment and cannot be easily removed
by natural processes, but tend to accumulate in the environment and in
the biota, causing biomagnification at higher trophic levels (Shilla et al.,
2019). This is a threat to the socio-economic status as well as the health
of the environment and can ultimately lead to the degradation of
important ecosystem services (Adams et al., 2019; Human et al., 2020).
In particular, Africa has gone through accelerated urbanisation and
industrialisation in the last five decades, which has increased pollution
pressure on marine environments (Sulla and Zikhali, 2018). Numerous
studies have researched the benefits of healthy wetland habitats (salt
marsh and seagrass) for the mitigation of the harmful effects of metal
pollution in estuaries (Caçador et al., 2009; Ali et al., 2013; Bonanno
et al., 2018). In light of local and global losses of salt marsh, it has
become increasingly important to conserve wetland plants considering
their contribution to estuarine health (Bornman et al., 2016; McOwen
et al., 2017). Knowing the capacity of wetland plant species to accu­
mulate metals is crucial in identifying suitable species for remediation
and increasing the importance of conserving local and endemic
vegetation.
Wetland vegetation occupies areas that naturally accumulate high
levels of metals and have developed physiological mechanisms to cope
with adversely higher concentrations (Wang and Liu, 2003; Weis and
Weis, 2004; Curado et al., 2014). The adaptation is in response to
photosynthetic performance, which can be reduced by competitive
metal ions, and include mechanisms such as the induction of metal
chelators, antioxidant defences, gene regulation, exclusion, ion ho­
meostasis and accumulation (Wiszniewska et al., 2018; Greco et al.,
2019). Whether phytoremediation is employed or not, wetland plants
can reduce the exposure of metals to the rest of the ecosystem, therefore,
reducing biomagnification and providing an important ecosystem ser­
vice (Windham et al., 2003; Weis and Weis, 2004; Reboreda and Caça­
dor, 2007; Curado et al., 2014). Phytostabilisation, where metals are
accumulated in the below-ground organs, is a very important strategy to
prevent metals from damaging toxic photosynthetic systems in the
shoots and leaves (Reboreda and Caçador, 2007). This strategy

* Corresponding author at: South African Environmental Observation Network (SAEON) Elwandle Coastal Node Nelson Mandela University, Ocean Sciences
Campus, 4 Gomery Avenue, Summerstrand, Gqeberha 6031, South Africa
E-mail address: lucienne.human@saeon.ac.za (L.R.D. Human).

https://doi.org/10.1016/j.marpolbul.2023.115007
Received 6 June 2022; Received in revised form 31 March 2023; Accepted 29 April 2023
Available online 15 May 2023
0025-326X/© 2023 Elsevier Ltd. All rights reserved.
M.A. Nel et al.
immobilises metals in more stable root biomass, and is a seasonal and
site-specific, long-term metal sink. Other strategies such as removal
(often employed by seagrasses) accumulate and nullify the toxic effects
of metals in the leaves, whereafter the leaves are dropped. This ensures
the survival of the individual plant but is more of a source than a sink of
metals, to the rest of the ecosystem and food chain. Compartmentali­
sation is a metal tolerance mechanism that involves metal uptake and
storage in various parts of plant organs, which lead to the negation of
metal toxicity in the plant and/or the rest of the environment (Soda
et al., 2012; Bonanno and Vymazal, 2017).
The aim of this study was to evaluate the seasonal compartmentali­
sation (leaves, shoots, roots) of potentially hazardous metals (Fe, Mn,
Zn, Cr, Pb, Ni, Cu) in salt marsh (Spartina maritima and Salicornia tege­
taria) and seagrass (Zostera capensis) plants in the Swartkops Estuary.
This estuary is densely developed with point sources originating from
urban (formal and informal) and industrial zones. The Swartkops Estu­
ary is an important socio-economic and environmental asset and has
high biodiversity importance in terms of fish, birds, and plants (Turpie
et al., 2002; Adams, 2020). Its intertidal salt marsh area is largely intact
with Salicornia tegetaria as a dominant species (Colloty et al., 2000).
Zostera capensis is endemic to Southern Africa, and little research on its
metal accumulation has been done. Therefore, it presents an important
opportunity to study the dynamics of compartmentalisation in these
plants. Higher capacity to accumulate metals will bring to light the
environmental value of salt marsh and seagrass, specifically in a densely
developed estuary such as the Swartkops Estuary, providing incentive to
conserve the habitats. This study will describe the patterns of accumu­
lation for metals in the selected plants and identify internal mobility
patterns and phytoremediation potential. We hypothesise that the salt
marsh (S. maritima and S. tegetaria) will have higher metal accumulation
than the seagrass (Z. capensis), and that compartmentalisation pathways
would be species-specific.
Fig. 1. Locations of the sampling sites in the Swartkops Estuary, Gqeberha, South Africa, (Site 1) Motherwell Canal, (Site 2) Markman Canal, (Site 3) Tiger Bay
Launch Site, (Site 4) Tipper’s Creek and (Site 5) Swartkops Estuary Mouth.
2
Marine Pollution Bulletin 192 (2023) 115007
2. Materials and methods
2.1. Study site
Sampling occurred at five different sites in the Swartkops Estuary, a
predominantly open estuary located on the warm-temperate south-east
coast of South Africa (Fig. 1) (van Niekerk et al., 2019). The sites were
located in the intertidal area of the middle and lower reaches of the
estuary. The freshwater inflow is restricted by damming and tributaries
that reduces freshwater inflow by 5 %, but input has increased overall
due to the Wastewater Treatment Works, which modified the natural
conditions of the estuary (Adams et al., 2019). It is well mixed and ex­
hibits highly stratified waters only during a high flood event. The salt
marshes cover an area of approximately 547.4 ha and is dominated by
S. maritima in the lower reaches. Salicornia sp. are found higher up the
intertidal gradient of the saltmarsh, while seagrass beds consisting of
Z. capensis is found along the subtidal-intertidal fringe. Swartkops Es­
tuary contains some of the largest Z. capensis beds (Adams et al., 2021).
These habitats are essential for nutrient cycling, fish nurseries and
aquatic habitat. The middle and lower reaches of the estuary is sur­
rounded by rural, urban, and industrial zones, which contribute to
effluent release into the estuary. The major industrial activities in the
vicinity include saltpans, sewage works, sand/clay mining, brickworks,
a power station, motor industry, wool industry, tanneries, extraction/
beneficiation processes, aquaculture, railway yards, depots, and some
agriculture (Binning and Baird, 2001). Previous studies in the water
column, sediments, and biota indicate increased concentrations of Cu,
Zn, Fe, Cd, As, Hg, and Pb in the ecosystem (Nel et al., 2015, 2020;
Phillips et al., 2015; Pretorius, 2015). The estuary is therefore subjected
to high anthropogenic pressure, with the potential to introduce metal
contaminants.
M.A. Nel et al.
2.2. Sampling
Sampling was carried out over a year period in autumn (30 April
2019), winter (29 July 2019), spring (28 October 2019) and summer (20
January 2020), during spring low tides. Sediment samples were
collected along the shore of the estuary within homogenous patches of
Z. capensis, S. maritima and S. tegetaria, at each of the sites. The plants
were randomly selected along a vertical line so that the three species
were within an intertidal range from each other. Samples were measured
in triplicate at each site, but sites were collated in this study, therefore
15 replicates were taken per plant, per season. Rhizosediment from the
surface (0–10 cm) of the plant was collected and contained within 50 ml
plastic jars with a screw cap, being careful not to disturb the root
biomass. A PVC corer (radius = 5.6 cm) was used to remove, whole plant
biomass from the selected plants, being careful to keep the roots intact
(depth = ca. 20 cm). The majority of the outer sediment was removed
from samples in situ, using a 1 mm wire meshed basket without dis­
turbing the integrity of the plant’s internal structure. Samples were then
transferred into sealable plastic ziplock packets. The samples were
transported in a cooled, thermally insulated cooler box and stored at
<5 ◦ C until further analysis, according to the specifications in (Four­
qurean et al., 2017). All containers and equipment used in handling the
samples were acid stripped with 1 % HNO3, rinsed with de-ionised water
and dried in their preparation.
Once in the laboratory, plant samples were washed gently under a
water jet with distilled water to remove any trapped particles, and then
rinsed with bi-distilled water to remove any other residues. The plants
were individually dissected into leaves, roots, and shoots, using stainless
steel scissors. Rhizomes were included into “roots”, while the tillers in
grasses were categorised under “shoots” (Caçador et al., 2009; Di Leo
et al., 2013). The individual plant organs were then freeze-dried
(Vacutec, V-FD12 Series Freeze Dryer) until a constant weight was
achieved. The plant sample then consisted of the same organs cut from
the same plant. Once dry, plant samples were weighed on a scale, and
ground in a stainless-steel coffee grinder and further with mortar-and-
pestle into a homogenous powder. Biomass values per area of the PVC
core (g m− 2) were also calculated with the radius of the corer at 5.6 cm.
Leftover residue in the grinders were cleaned between each sample with
brushes and paper cloths. Sediment samples were similarly freeze-dried
until a constant weight, prior to the metal extractions.
2.3. Analysis of metal concentrations through TXRF spectrometry
The single extraction entailed a conventional overnight wet diges­
tion method (Du Laing et al., 2003; Phillips et al., 2015), to extract the
total metal concentration in plant material and sediment. The sediment
samples and dissected plant samples were digested for the measurement
of Cr, Cu, Fe, Mn, Ni, Pb, and Zn. An aliquot of freeze-dried sediment
first was pushed through a 1 mm sieve to remove any unwanted mate­
rial. The homogenous plant and sediment samples (1.0000 ± 0.001 g)
were left in 10 ml HNO3 (A.R grade, 65 %, p.a.) overnight, and then
refluxed on a hotplate for ~5 h at 110 ◦ C, using a watch-glass, until the
production of NO2 fumes had ceased. After evaporation to near dryness
the sample was diluted with 20 ml of 2 % (v/v H2O) nitric acid and
transferred to 50 ml volumetric flasks after filtering (Whatman™ GF/F
Filters) to remove particulates and diluted to the mark with ultra-pure
water (Milli-Q®, Direct 8 Water Purification System).
Extractants were analysed for the selected metals using a Total
Reflection X-ray Fluorescence Spectrometer (TXRF, Bruker, S4 T-star),
an alternative to the ICP-OES (Inductively Coupled Plasma-optical
emission spectrometer) spectrometer. The TXRF is a reliable method
of analysis, which is very efficient at using small amounts of sample
(Vilhena et al., 2021). The spectrometer is equipped with a molybdenum
(Mo) and tungsten-brems (W) x-ray tube. A 10 μl mixture of the
following was pipetted in the middle of the circular sample carriers
(polished quartz glass): (1) 7000 μl of the digested sample, (2) 70 μl of
3
Marine Pollution Bulletin 192 (2023) 115007
the 100 ppm internal standard(s) i.e., 1 mg L− 1, and 70 μl of 0.3 g L− 1
poly(vinyl) alcohol solution (for sample homogeneity). Sample carriers
were prepared according to the indication of the manufacturers,
including the siliconization of the sample carriers with hexamethyldi­
silazane (≥ 99 %, MerckSA) to make the surface hydrophilic. Regarding
internal standards, Ga and Y were used for plant samples and Se was
used for sediment samples. Internal standards were chosen that do not
naturally occur in sediment or plant material, and that corresponds with
the wavelengths of the measured metals. The internal standard method
also corrects for variations in sample deposition on the reflective sample
carriers (Montero Alvarez et al., 2007). The stocked sample carrier was
then transferred to a hotplate and maintained at low heat (ca. 55 ◦ C)
until the sample was dry. The TXRF analyses the surface to detect the
metal concentrations using x-ray fluorescence. The metals are identified
based on their fluorescence Kα lines (peaks). The instrument was cali­
brated according to manufacture indications before each analysis, which
entails alignment, and calibration of concentrations using a pre-set
standard. Scan time was defined at 1000 s. Blanks runs were per­
formed alongside the samples, which consisted of the HNO3 acid, the
poly(vinyl) alcohol solution, and the internal standard. The detection
limits (mg L− 1) for the selected metals (Cr = 0.005, Cu = 0.001, Fe =
0.004, Mn = 0.003, Ni = 0.001, Pb = 0.001, Zn = 0.001) were provided
by the spectrometer. It is calculated with the area of the target elemental
peak and the area of the adjacent (background) peaks (Eq. (1)) (Matu­
siak et al., 2017). Percentage recovery of metals were calculated by
comparing the detected concentrations of certified reference materials
(Sigma-Aldrich, Metals in Sewage Sludge, SQC001S-30 g and Institute for
Reference Materials and Measurements, Rye Grass, ERM® - CD281).
√̅̅̅̅̅̅̅̅
3.Ci. NBG
LLDi = (1)
Ni
Where: LLDi = the lowest limit of detection of the element “i” [PPM], Ci
= concentration of element “i” in the measured sample [PPM], NBG =
background area subjacent to the element “i” Kα lines, and Ni = peak
area of the element “i” Kα lines.
2.4. Determining metal compartmentalisation in macrophytes
Bioconcentration and translocation factors were calculated to obtain
information on plant bioaccumulation and compartmentalization. Both
factors were calculated as a ratio between the total metal concentrations
of the leaves, shoots, roots, and the sediment collected within the
rhizosphere. The factors can be expressed as follows:
Bioconcentration Factor (BCF) = Croots /Csediment
Translocation Factors (TF) = Cleaf /Croots
Where Croots,
= CShoot /Croots
= Cleaf /CShoot
Cshoot, Cleaf, and Csediment are the concentrations of the specific element
in the roots, shoots, leaves, and the sediment (mg kg− 1, dry weight),
respectively. The BCF expresses the efficiency of a plant to take up and
accumulate the element in its roots, and a larger value indicates a higher
accumulation ability (Soda et al., 2012). The TF expresses the specific
element mobility within the plant, and a larger value indicates a higher
translocation ability (Deng et al., 2004). A BCF >1 has been argued to be
a great candidate for phytostabilisation (Usman et al., 2012).
2.5. Statistical analysis
Each statistical test was accompanied by 15 replicate samples. The
analyses of variance (ANOVA, random effects) were used to test for
differences in all the data. Tukey’s post hoc test was then used to
compare the means where significant differences were found. Differ­
ences were significant when the p-value was <0.05. The assumption of
normality was verified before analysis with the Shapiro-Wilks test,
accompanied with the respective histograms, to test for skewness.
M.A. Nel et al. Marine Pollution Bulletin 192 (2023) 115007

Variables were log transformed if the null hypothesis for normality was
rejected. Once the BCF and TF were calculated, outliers were removed
from these compartmentalisation ratios using box-and-whisker plots.
The subsequent results were graphically plotted. Metal stocks (μg m− 2)
were calculated by multiplying the biomass per unit area (g m− 2) with
the metal concentrations (μg g− 1 of sediment), and then graphically
displayed. All statistical processing and graphical display were per­
formed in RStudio (3.5.1/4.0.2; RProject) using the packages Vegan,
RColorBrewer, Tidyverse, R-commander, mvabund and Geoveg, and
Microsoft Excel (365).
3. Results
3.1. Metal concentrations in plants and rhizosediment
The average metal concentrations in the rhizosediment for all five
sites, showed the following patterns in descending order: Fe > Mn > Zn
> Cr > Pb > Ni > Cu and S. tegetaria > S. maritima ≥ Z. capensis
(Table 1). Furthermore, no significant statistical differences were found
between the seasons for a specific metal and rhizosediment type (p >
0.005).
The studied species generally showed different metal concentrations
with different trends for the elements (Table 2). Spartina maritima con­
tained the highest Cr and Ni, followed by S. tegetaria and then Z. capensis.
Zostera capensis, contained the most Cu, Mn, and Zn. Salicornia tegetaria
contained the second highest Mn and Cu, but the least amount of Zn. The
two metals, Fe and Pb, showed similar concentrations between the
whole plant species. Metal concentrations in plant tissues generally
showed consistent trends. Spartina maritima and S. tegetaria had their
highest concentrations in the below-ground organs, indicating a general
trend of root > shoot > leaf, while Z. capensis showed the opposite, with
a general trend of leaf > root > shoot or leaf ≥ root > shoot.
There weren’t many general trends regarding metal concentrations
in the leaf tissues. For example, leaf tissue of S. maritima showed the
general trend winter ≥ summer > spring > autumn, but it was not sig­
nificant for any element. Metal concentrations in the leaf tissue of
S. tegetaria decreased from the winter to autumn to summer or spring,
respectively. Apart from Mn, no particular seasonal trend was observed
for the leaf tissue of Z. capensis. Higher Mn concentrations occurred in
summer compared to winter, and higher spring concentrations
compared to winter. The winter concentrations in the leaves were in
general higher than the other seasons. In most cases only the highest and
lowest concentrations within a plant species were statistically
significant.
Regarding seasonality in shoot tissue concentrations — this did not
show many comparable trends between plants and the specific metals.
For S. maritima winter metal concentrations were also generally the
highest, with the exception of Cr and Ni (in autumn) and Pb (in sum­
mer). These exceptions were not statistically significant (p > 0.05).
Similarly, the leaf tissue in Z. capensis and shoot concentrations were
generally higher in spring, and the exceptions were Mn (in summer) and
Zn and Cu (in autumn). Regarding S. tegetaria shoot tissues: Fe was
highest in spring; Mn, Zn, and Cr highest in autumn; and Pb, Ni, and Cu
highest in summer. The latter elements can be contrasted with Fe, Mn,
Zn and Cr, where summer concentrations were at its lowest.
Lastly, seasonality in root concentrations showed differences for
S. tegetaria and Z. capensis, but S. maritima concentrations showed little
seasonal differences. Spring concentrations were generally the highest
between seasons for Z. capensis and S. tegetaria, and the lowest in sum­
mer. Some exceptions for Z. capensis include Zn, where winter concen­
trations were the highest and Ni, where autumn had the lowest
concentration. For S. tegetaria, Cr, Ni, and Cu showed a different trend,
as autumn was the highest for the two former elements and winter was
higher for the latter element. Spartina maritima root tissue concentra­
tions were similar seasonally.
3.2. Translocation and bioconcentration
Firstly, TF(leaf/root) ratios showed that S. tegetaria and S. maritima
values were generally <1 (Fig. 2). Zostera capensis TF(leaf/root) were
higher than 1 for all metals except for Fe, and therefore was very
different compared to the other plants. Regardless, the TF(leaf/root) had
the following mean pattern: Mn > Zn > Cu > Cr > Ni = Pb > Fe, where
“>” is significantly different at p < 0.05. Salicornia tegetaria and

Table 1
1
Average metal concentrations (μg g− dry weight ± SD) in the rhizosediment of Z. capensis, S. maritima, and S. tegetaria, for each season in all five sites (n = 15).
− 1
μg.g Z. capensis S. maritima S. tegetaria

Fe Autumn 8496.7 ± 4455.7 10,746.5 ± 6407.8 15,698.1 ± 11,063.9

Winter 7806.6 ± 5020.2 10,899.3 ± 9064.0 14,524.2 ± 9860.5
Spring 5589.4 ± 2705.1 6470.2 ± 4002.0 10,919.4 ± 6099.4
Summer 5617.9 ± 2068.0 11,544.4 ± 6115.4 13,737.5 ± 8785.6
Mn Autumn 69.7 ± 46.6 91.2 ± 55.9 223.5 ± 176.6
Winter 50.1 ± 21.0 84.0 ± 60.9 222.7 ± 140.5
Spring 39.6 ± 10.8 49.2 ± 26.9 165.9 ± 102.5
Summer 39.9 ± 13.2 126.6 ± 127.4 172.0 ± 118.4
Zn Autumn 45.7 ± 20.6 45.1 ± 18.2 68.7 ± 39.3
Winter 59.8 ± 23.4 63.8 ± 29.0 73.4 ± 39.7
Spring 29.4 ± 10.9 35.5 ± 14.1 44.5 ± 20.6
Summer 32.4 ± 9.9 48.1 ± 18.2 60.7 ± 33.9
Cr Autumn 26.3 ± 10.5 22.0 ± 8.2 31.9 ± 25.9
Winter 21.2 ± 11.6 25.4 ± 17.2 29.9 ± 24.1
Spring 15.1 ± 5.9 18.4 ± 9.7 19.8 ± 9.61
Summer 16.9 ± 5.1 27.9 ± 13.5 29.3 ± 16.9
Pb Autumn 12.2 ± 8.4 11.9 ± 8.4 24.2 ± 22.7
Winter 11.7 ± 10.7 16.6 ± 16.4 25.1 ± 22.9
Spring 6.9 ± 3.2 9.3 ± 5.3 13.5 ± 8.7
Summer 7.6 ± 2.6 15.8 ± 9.4 21.7 ± 16.8
Ni Autumn 6.3 ± 2.9 8.4 ± 4.2 12.2 ± 7.6
Winter 7.1 ± 3.3 7.6 ± 5.6 10.9 ± 7.2
Spring 4.5 ± 3.0 5.7 ± 2.8 8.1 ± 4.4
Summer 4.9 ± 3.6 9.3 ± 2.5 11.6 ± 6.6
Cu Autumn 5.7 ± 3.3 5.8 ± 3.6 9.6 ± 6.2
Winter 5.2 ± 4.1 6.1 ± 4.9 7.6 ± 5.4
Spring 3.9 ± 2.0 4.5 ± 2.4 6.4 ± 4.2
Summer 3.6 ± 1.2 6.5 ± 3.3 9.4 ± 7.4

4
 M.A. Nel et al.
Table 2
1
Average metal concentrations (μg g− dry weight ± SD) in leaves, shoots, and roots of Z. capensis, S. maritima, and S. tegetaria, for each season (n = 15).
− 1
μg.g Z. capensis S. maritima S. tegetaria

Leaf Shoot Root Leaf Shoot Leaf Shoot

Fe Autumn 2673 ± 349a,1 569 ± 74a,b,2 4360 ± 464a,1 822 ± 107a,1 1403 ± 166a,b,1 727 ± 86a,b,1 1410 ± 129a,1
Winter 3232 ± 1114a,1 1180 ± 197a,2 8031 ± 1009a,b,3 1190 ± 140a,1 2922 ± 906a,1 1126 ± 170a,1 1108 ± 135a,1
Spring 3987 ± 520a,1 2003 ± 329a,b,1 14,079 ± 1705b,2 955 ± 139a,1 1060 ± 113a,b,1 514 ± 94a,b,1 1684 ± 298a,2
Summer 1734 ± 202a,1 486 ± 82b,2 3719 ± 458a,1 631 ± 79a,1 749 ± 73b,1 377 ± 40b,1 853 ± 150a,1
Mn Autumn 1566 ± 288a,b,1 206 ± 30a,2 198 ± 31a,2 33.5 ± 3.9a,1 43.9 ± 7.2a,1,2 43.3 ± 5.3a,1 71.3 ± 9.5a,1,2
Winter 1034 ± 135a,1 215 ± 47a,2 269 ± 48a,2 48.3 ± 6.0a,1 72.0 ± 13.6a,1 52.2 ± 10.4a,1 47.0 ± 6.5a,1
Spring 2012 ± 279a,b,1 180 ± 17a,2 297 ± 30a,2 47.8 ± 5.1a,1 66.0 ± 17.7a,1 24.7 ± 3.6a,1 53.2 ± 7.9a,1
Summer 2749 ± 397b,1 318 ± 59a,2 341 ± 31a,2 29.2 ± 4.5a,1 41.5 ± 6.3a,1,2 22.9 ± 3.0a,1 36.8 ± 3.0a,1,2
Zn Autumn 164.8 ± 11.0a,1 113.5 ± 9.1a,1,2 87.7 ± 14.3a,b,2 39.3 ± 4.5a,1 45.3 ± 4.1a,b,1 38.3 ± 4.2a,1 52.4 ± 4.8a,1
Winter 187.9 ± 11.6a,1 88.7 ± 4.7a,2 101.8 ± 8.4a,2 48.2 ± 3.9a,1 84.2 ± 8.5b,2 49.0 ± 3.7a,1 49.5 ± 3.9a,1
Spring 151.4 ± 8.4a,1 67.8 ± 4.4a,2 92.1 ± 8.6a,b,2 36.4 ± 2.3a,1 38.8 ± 2.3a,b,1 33.7 ± 3.3a,1 41.5 ± 4.0a,1
Summer 129.4 7.7a,1 69.4 4.3a,2 56.5 4.5b,2 44.6 3.2a,1 54.2 6.9b,1 38.0 4.6a,1 40.2 3.9a,1
5

± ± ± ± ± ± ±
Cr Autumn 19.0 ± 4.6a,1 4.6 ± 4.0a,2 6.1 ± 0.7a,c,1,2 15.5 ± 2.6a,1 64.7 ± 11.8a,2 6.7 ± 0.7a,1 8.0 ± 0.9a,b,1
Winter 25.9 ± 16.9a,1,2 10.4 ± 5.5a,1 23.7 ± 5.8a,b,2 16.8 ± 3.5a,1 50.3 ± 17.4a,b,1,2 5.5 ± 1.1a,1 7.2 ± 1.9a,c,1
Spring 12.1 ± 1.2a,1 26.4 ± 10.9b,1 37.2 ± 5.6b,1 10.4 ± 2.1a,1 17.1 ± 2.5b,1 1.8 ± 0.4b,1 4.0 ± 0.6b,c,1
Summer 7.4 ± 1.3a,1 5.8 ± 2.2a,b,1 6.4 ± 1.3c,1 6.8 ± 1.0.a,1 14.4 ± 2.2b,1 1.3 ± 0.4b,1 1.4 ± 0.2c,1
Pb Autumn 13.9 ± 1.5a,1 4.0 ± 0.2a,2 9.7 ± 1.3a,b,1,2 2.2 ± 0.2a,1 2.9 ± 0.3a,1 3.0 ± 0.3a,1 4.0 ± 0.2a,1
Winter 6.6 ± 1.1b,1,2 3.9 ± 0.8a,1 10.4 ± 1.8a,b,2 2.8 ± 0.6a,1 5.1 ± 1.8a,b,1 3.4 ± 0.3a,1 3.0 ± 0.2a,1
Spring 12.9 ± 1.0a,1 5.5 ± 0.3a,2 16.8 ± 1.5a,1 2.5 ± 0.2a,1 2.9 ± 0.2a,1 1.9 ± 0.2a,1 5.3 ± 1.1a,2
Summer 9.0 ± 0.7a,b,1 3.4 ± 0.2a,2 5.8 ± 0.5b,1,2 5.5 ± 0.1b,1 6.4 ± 0.3b,1 2.0 ± 0.2a,1 11.1 ± 7.5a,1
Ni Autumn 10.6 ± 1.3a,1 6.9 ± 1.0a,1 5.3 ± 0.7a,1 7.6 ± 1.1a,1 28.7 ± 3.6a,2 4.6 ± 0.4a,1 5.1 ± 0.4a,1
Winter 14.3 ± 6.7a,1 7.9 ± 2.6a,1 11.2 ± 2.2a,b,1 8.1 ± 1.5a,1 23.2 ± 7.7a,b,1 4.3 ± 0.6a,c,1 5.0 ± 0.8a,b,1
Spring 9.0 ± 1.0a,1 14.8 ± 5.2a,1 17.1 ± 2.8b,1 5.2 ± 0.8a,1 8.8 ± 1.1b,1 1.8 ± 0.2b,c,1 3.1 ± 0.3a,b,1
Summer 6.5 ± 0.7a,1 4.5 ± 1.0a,1 4.6 ± 0.6a.b,1 3.7 ± 0.4a,1 7.6 ± 1.0b,1 1.1 ± 0.1b,1 13.6 ± 11.8b,1
Cu Autumn 64.6 ± 7.1a,1 32.9 ± 4.0a,1 42.0 ± 6.5a,b,1 6.0 ± 0.8a,1 6.3 ± 0.6a,1 9.8 ± 1.1a,1 17.9 ± 2.7a,1
Winter 56.6 ± 4.3a,1 27.1 ± 2.2a,2 51.3 ± 3.8b,1,2 9.4 ± 1.1a,1 13.2 ± 2.6b,1 20.4 ± 2.7b,1 18.5 ± 1.7a,1
Spring 59.0 ± 3.8a,1 23.2 ± 1.7a,2 56.9 ± 4.1b,1 5.9 ± 0.5a,1 5.6 ± 0.6a,b,1 6.4 ± 0.4a,b,1 15.0 ± 1.5a,1
Summer 38.3 ± 3.5a,1 18.0 ± 1.0a,1 21.7 ± 1.7a,1 6.4 ± 0.6a,1 7.9 ± 1.3a,b,1 10.8 ± 1.7a,b,1 20.2 ± 8.2a,1

Marine Pollution Bulletin 192 (2023) 115007

M.A. Nel et al. Marine Pollution Bulletin 192 (2023) 115007

Fig. 2. Average seasonal [leaf/root] Translocation Factor for Z. capensis, S. maritima, and S. tegetaria where; (a) Fe, (b) Mn, (c) Zn, (d) Cr, (e) Pb, (f) Ni, and (g) Cu (N
= 15).

S. maritima deviated from the mean and showed Zn > Cu = Mn > Ni =
Pb > Cr = Fe and Zn = Mn > Cu > Pb = Ni > Cr = Fe, respectively (“>”
= significant; p < 0.05). Note that the lowest four metals (Cr, Ni, Pb, Fe)
and the highest three metals (Mn, Zn, Cu) do not change between spe­
cies. Seasonal differences were only prominent for Z. capensis, where a
decrease in the TF(leaf/root) occurred from autumn to winter followed by
a similar translocation in spring and a relatively higher translocation in
summer.
In general, TF(leaf/shoot) (Fig. 3) acted similarly to TF(leaf/root), in that
S. maritima and S. tegetaria ratios were generally below 1, and had only a
few seasonal differences. That is, autumn and winter generally showed
higher translocation for S. tegetaria, and spring and summer showed
higher translocation for S. maritima. However, these observations were
not significant. Any trends concerning the specific metals were also not

Fig. 3. Average seasonal [leaf/shoot] Translocation Factor for Z. capensis, S. maritima, and S. tegetaria where; (a) Fe, (b) Mn, (c) Zn, (d) Cr, (e) Pb, (f) Ni, and (g) Cu
(N = 15).

6
M.A. Nel et al.
that apparent for S. maritima or S. tegetaria, as the ratios were similar to
each other. In most cases, the Z. capensis TF(leaf/shoot) far exceeded those
of the other two plant species. Moreover, the TF(leaf/shoot) was more
variable between elements, and occurred as follows, in descending
order: Mn > Fe > Pb = Cu > Zn > Ni = Cr. It also showed more seasonal
variability. Seasonal differences for Z. capensis were different in this
case, between metals. Winter values were the lowest for Fe, Mn, and Pb,
while high values in autumn were found for Fe, Cr and Pb. The TF(leaf/
shoot) was comparably much higher in Z. capensis than the TF(leaf/root).
A different trend was observed for the TF(shoot/root) (Fig. 4) compared
to the other translocation factors. The range of values were much lower,
as seen on the figures, where only Z. capensis ratios exceeded 1 in a few
seasons for Zn and Ni. The translocation from root to shoot remained the
lowest for S. tegetaria and S. maritima, but Z. capensis values were much
lower and show similarity for Fe, Mn, Zn, Cr, Ni, and Cu. Zostera capensis
had higher translocation of certain metals (Mn, Zn, Cr, Pb, and Ni) in
autumn and summer, compared to the other plants in the same season.
The mean TF(shoot/root) had the following order in terms of metals:
Z. capensis̶ Zn > Ni = Mn > Cu > Cr = Pb > Fe, S. maritima – Zn = Mn >
Ni > Cr > Cu = Pb > Fe, and S. tegetaria – Zn > Cu = Mn > Pb = Ni > Fe
> Cr.
The BCF(root/sediment), that is translocation from sediment to roots
(Fig. 5), generally show lower Z. capensis values compared to the other
plants. Exceptions occurred for Mn and Zn, where Z. capensis BCF(root/
sediment) was higher or similar to the other plant species, respectively. On
average, it exceeded a value of 1 for all plants. The mean BCF(root/sedi­
ment) for each plant species had the following order: Zostera capensis –
Mn > Zn ≥ Ni ≥ Cu > Cr ≥ Fe ≥ Pb, S. maritima – Cu > Ni ≥ Cr > Zn ≥
Pb ≥ Mn ≥ Fe, and S. tegetaria – Cu > Mn ≥ Ni ≥ Zn ≥ Cr ≥ Pb ≥ Fe.
Seasonal differences were different for each plant and metal. In spring,
Fe concentrations were highest for all three plants species. Zostera
capensis had higher BCF(root/sediment) in spring, for all cases except Mn.
Spartina maritima also displayed a higher BCF in spring, except for Cr, Ni,
and Cu. Salicornia tegetaria also had higher BCF(root/sediment) in spring for
Fe, Mn, Zn, and Pb, but other metals (except Cu) showed similar values
in autumn and spring.
Fig. 4. Average seasonal [shoot/root] Translocation Factor for Z. capensis, S. maritima, and S. tegetaria where; (a) Fe, (b) Mn, (c) Zn, (d) Cr, (e) Pb, (f) Ni, and (g) Cu
(N = 15).
7
Marine Pollution Bulletin 192 (2023) 115007
3.3. Plant biomass
Total plant biomass (Table 3; g m− 2) for S. maritima (e.g., Autumn =
3440 g m− 2) was more than double compared to S. tegetaria (e.g.,
Autumn = 1001 g m− 2), which in turn was more than double the total
biomass of Z. capensis (e.g., Autumn = 328 g m− 2). Zostera capensis alone
showed seasonal variation in total biomass, where summer had signifi­
cantly higher total biomass compared to autumn and spring. The plant
compartments (leaf, shoot, root) follow the same pattern as the total
biomass, that is: S. maritima > S. tegetaria > Z. capensis. They showed
different seasonal variations within a plant species. Leaf biomass was the
most dynamic between seasons. Zostera capensis leaf biomass was
highest in summer, which was significant to autumn (p < 0.0001) and
spring (p = 0.0002). Salicornia tegetaria and S. maritima leaf biomass
were highest in spring, but it was only significant for S. tegetaria. Shoot
biomass did not differ seasonally for S. maritima and Z. capensis. How­
ever, S. tegetaria had significantly lower shoot biomass in autumn (p <
0.0006), only. High root biomass could be seen in autumn for S. maritima
(2147 ± 347 g m− 2) compared to the lowest in summer (1246 ± 121 g.
m− 2). The lowest root biomass was also in summer for S. tegetaria (412
± 54 g m− 2) and Z. capensis (104 ± 9 g m− 2).
3.4. Metal stocks in plants
Metal stocks (Fig. 6; μg m− 2) showed similar trends between metal
elements. They were generally lower in S. maritima, while Z. capensis
and S. tegetaria had comparable metal stocks. Salicornia tegetaria and S.
maritima stocks were lowest in spring compared to its highest in winter.
Zostera capensis showed a decline in metal stock from autumn to sum­
mer. However, some metals like Mn had comparable concentrations
between winter and spring.
4. Discussion
4.1. Compartmentalisation
The studied species show different metal concentrations (Table 2),
M.A. Nel et al. Marine Pollution Bulletin 192 (2023) 115007

Fig. 5. Average seasonal [root/sediment] Bioconcentration Factor for Z. capensis, S. maritima, and S. tegetaria where; (a) Fe, (b) Mn, (c) Zn, (d) Cr, (e) Pb, (f) Ni, and
(g) Cu (N = 15).

Table 3
2
Average dry biomass (g m− ± SE) for plant organs and whole plant of Z. capensis, S. maritima, and S. tegetaria in the Swartkops Estuary (n = 15).
Biomass Z. capensis S. maritima S. tegetaria
− 2
g.m

Leaf Autumn 171 ± 22 369 ± 43 406 ± 28

Winter 363 ± 65 473 ± 78 433 ± 74
Spring 216 ± 31 541 ± 38 643 ± 47
Summer 472 ± 53 460 ± 53 426 ± 34
Shoot Autumn 30 ± 3 924 ± 80 86 ± 8
Winter 52 ± 11 1016 ± 165 233 ± 23
Spring 34 ± 4 1273 ± 106 258 ± 20
Summer 52 ± 7 1104 ± 83 161 ± 14
Root Autumn 128 ± 7 2147 ± 347 509 ± 51
Winter 160 ± 18 1254 ± 188 491 ± 94
Spring 150 ± 18 1625 ± 146 648 ± 132
Summer 104 ± 9 1246 ± 121 412 ± 54
Total Autumn 328 ± 25 3440 ± 378 1001 ± 66
Winter 575 ± 84 2743 ± 370 1156 ± 121
Spring 401 ± 39 3439 ± 202 1549 ± 168
Summer 629 ± 56 2810 ± 203 999 ± 78

S. maritima and S. tegetaria had higher metal concentrations in their roots
compared to their leaves, while metals in Z. capensis was the highest in
the leaves. Seasonal patterns were also unique to the plant species and
the metal in question. Metal accumulation patterns are generally not
comparable between life forms or genera and is species specific
(Bonanno et al., 2018). The life forms of the three plant species are very
different i.e., S. maritima is a grass, Z. capensis is a seagrass, while
S. tegetaria is a succulent. The highly dense root structure of S. maritima
compared to Z. capensis and S. tegetaria may contribute to its success as
an accumulator. The highest metal concentrations in leaves did gener­
ally not occur in the growing season but was in autumn and winter.
Dilution would be a key contributing plant characteristic to consider. It
can occur in the above-ground organs during the growing seasons as
plants get ready to reproduce (Weis and Weis, 2004). This is when rapid
growth of biomass dilutes the metal concentrations in the plant. Con­
centrations in young tissues have been found to be much lower in
Spartina alterniflora, and other salt marsh plants (Weis and Weis, 2004).
However, root and shoot concentrations were generally highest in spring
or summer, the growing seasons. Ragsdale and Thorhaug (1980) found
that in several salt marsh and seagrass distributed along the coast of the
United States of America, Cd, Cr and Mn increased at the end of the
growing season, while other metals (Pb, Zn, Cu) remained seasonally
constant. They attributed these differences to plant physiology and
senescence. To get rid of the metal burden plants concentrated metals at
the end of the growing season when plants senesced from living to
standing dead litter. Other evidence provided by Walsh and Grow
(1973) indicated that in Thalassia testudinum Mn, Fe and Zn varied
seasonally in the leaf fraction but had little variation in rhizome fraction.
This was related to a change in protein content which was higher in the
leaf fraction compared to the rhizomal fraction. The studied plants in the
Swartkops Estuary produced the most statistically different metal con­
centrations in plant organs, in summer, indicating the aforementioned

8
M.A. Nel et al.
2
Fig. 6. Average whole plant stock (μg m− ± SE) for Z. capensis, S. maritima, and S. tegetaria in each season, where (a) Fe, (b) Mn, (c) Zn, (d) Cr, (e) Pb, (f) Ni, and (g)
Cu (n = 15).
strategies in response to the growing season. However, other studies
have found that well-defined seasonality regarding metal concentrations
in roots is difficult to observe (Caçador et al., 2009; Windham et al.,
2003).
There is a large difference between the metal TF (Figs. 2, 3 and 4) of
S. maritima, S. tegetaria, and Z. capensis. The latter showed >10 times
translocation, indicating the transfer of metals to the above-ground
biomass of the plant. The metals that contained similar TF of root to
leaf, and shoot to leaf included Mn, Zn, Cr, and Ni — suggesting similar
rates of transfer between these compartments. As was the case for Cd,
Co, Mn and Zn for Z. marina (Lee et al., 2019). The other three metals Pb,
Fe, and Cu showed large differences between TF for Z. capensis. The non-
essential metal, Pb, showed lower TF from root to the shoot, and higher
TF from shoot to leaf, which indicates limited uptake of this metal, but
high sequestration in the leaves once it is taken up by the plant. Lead
toxicity in Z. marina is known to cause higher shoot density, which
would promote translocation to the leaves, and lower chlorophyll values
(Hoven, 1998). Zostera capensis had similar Pb concentrations in the
leaves and roots, which is what was found for Z. marina by Lyngby and
Brix (1984) and Lee et al., (2019) in meadows from Limfjord, Denamark
and Koje Bay, Korea, respectively. The low Fe TF (from roots to above-
ground organs) in this study indicates low uptake but high translocation
to the leaves, indicative of its essentiality for the production of chloro­
phyll. The uptake of Fe is highly regulated in these plants, due to the low
solubility (bioavailability) of Fe in sediments (Guerinot and Ying, 1994).
The TF of Cu showed the same pattern as Fe, with high translocation to
the leaves, but lower translocation to the shoots.
There were low TFs recorded for S. tegetaria and S. maritima, spe­
cifically from root to shoot and root to leaf (Figs. 2, 3 and 4). Trans­
location of metals to above-ground organs were minimal. These two
plant species also had very similar TFs. The turn-over of above-ground
organs (leaves and shoots) are generally higher than below-ground or­
gans and are more likely to be exported out of the marsh, enriching other
areas with metals carried within them. The negligible TF, high BCF
(Fig. 6) and the statistically stable leaf biomass (Table 3) found in these
plants make them ideal candidates for phytostabilisation. These char­
acteristics indicate that their role in the metal budget in the sediment is
very low (Duarte et al., 2010), as the metals in the plants are unavailable
for uptake by other organisms. The main organ of bioaccumulation in
S. tegetaria and S. maritima was the roots, since BCF was much higher in
9
Marine Pollution Bulletin 192 (2023) 115007
comparison to the TF. The work of Burke et al. (2000) showed that
S. alterniflora from a marsh in Hackensack Meadowlands, USA, was able
to actively excrete Cr, Cu, Pb and Zn through its salt glands, and may be
another strategy used by S. maritima to maintain a low concertation of
metals in its above ground biomass. Root accumulation of metals is a
common strategy in salt marsh plants (Bonanno and Vymazal, 2017;
Deng et al., 2004; Gedan et al., 2009; Reboreda and Caçador, 2007b).
The compartmentalisation strategy employed by these two plants pre­
vents toxic levels of metal accumulation in the photosynthetic organs
(Bonanno et al., 2017; Phillips et al., 2015; Weis and Weis, 2004). This
study showed that all three plant species, S. tegetaria, S. maritima and
Z. capensis, are good bioaccumulators, by having high BCF. The
following metals bioaccumulated (BCF > 1): (1) Z. capensis for Mn, Zn,
and Ni, (2) S. tegetaria for Mn, Zn, Cr, Pb, Ni, and Cu, and (3) S. maritima
for Fe, Zn, Cr, Pb, Ni, and Cu.
Seasonal metal bioconcentration to the roots were generally highest
in the spring/summer months for the selected wetland plants. The
warmer seasons increases the physiological requirements of plants as
they prepare for growth and reproduction (Yruela, 2009). Lead is a non-
essential metal and also showed this maximum in the growing seasons.
However, Cr, Ni, and Cu BCF-maximums were also found in autumn. A
decrease in metal uptake activity in the autumn and winter coincide
with the reduction in metabolic activity during the colder months (Feng
et al., 2017). The Swartkops Estuary is a warm-temperate estuary and
the plants do not fully go dormant in the colder months, which is what
was found for the warm-temperate Tagus Estuary in Portugal (Caçador
et al., 2000). Nevertheless, these past studies still found a reduction in
uptake in a response to colder temperatures. Translocation of the metals
to the leaves were low in the salt marsh plants (S. tegetaria and
S. maritima) and did not reflect seasonal growing requirements. Salt
marsh plants generally restrict metal uptake through the excluder
strategy to the surrounding sediment, thereby preventing translocation
to above ground tissue. This was found for S. alterniflora and Scirpus
mariqueter growing in the Dongtan wetland China (Quan et al., 2007).
Zostera capensis translocation to the leaves indicate metal growth re­
quirements towards chlorophyll production for the warmer seasons (Zn,
Cu, Mn), but also displayed high TF (Fe, Ni, Cr, Pb) in preparation for the
colder winter season. Weis and Weis (2004) indicated that younger
leaves in seagrasses, formed during the growing seasons, contain less
metal concentrations, but that translocation occurs into older leaves to
M.A. Nel et al.
get rid of metal loads in the plant during leaf senescence in colder
months.
4.2. Metal stock
Wetland plants are excellent sinks of metals, accumulating high
concentrations in their tissues. Their potential as a metal sink can be
tested with the level of metals stored in a specific area, relative to the
density that the plants occur in. This study found that the metal stock of
Z. capensis were generally lower than S. tegetaria and S. maritima, even
though the concentrations of metals were the highest. This can be
attributed to the relatively high biomass (g m− 2) found for this plant
species (Table 3; Fig. 6). Spartina maritima is well-known for excellent
accumulation of metals and for its potential in ecological engineering
through phytoremediation (Curado et al., 2014; Williams et al., 1994).
The biomass (g m− 2) of the plants followed the general order of
S. maritima > S. tegetaria > Z. capensis, and was most seasonally variable
for the seagrass, Z. capensis. The growing season of this seagrass occurred
in summer. Higher seasonal variability in the leaf biomass of Z. capensis
indicate high rates of leaf cycling, and the formation of leaf litter. Leaf
litter increases the mobility of metals in the sediment as they decompose
(Helfield and Diamond, 1997), after which they are available for uptake
by the plants again. In very hydrologically dynamic areas leaf litter can
promote the export of metals to other areas of the estuary. Caçador et al.
(2004) found that nearly 86 % of the produced above-ground plant
material was exported to other areas of the Tagus Estuary, Portugal.
Duarte et al. (2017) indicated that in the Tagus Estuary outwelling to the
ocean will increase due to sea level rise and climatic change — the
detritus is generally retained at the source during the neap tide and
exported during spring tide. The wetlands in the middle reaches of the
Swartkops Estuary (Site 3 and Site 4) are conducive to trapping mate­
rials from upstream sites. The formation of Z. capensis leaf litter likely
occurred in autumn and to a lesser extent in spring, as these seasons had
the lowest leaf biomass (Table 3). The results suggest that Z. capensis
may employ a metal tolerance strategy to remove toxic levels of metals
from the plant by translocating metals to the leaves and then shedding
them. This strategy is called the “removal” strategy. Metals accumulated
in the above-ground tissue can cause toxic effects, shedding of these
parts can prevent the death of the individual plant (Weis and Weis,
2004). Bonanno and Borg (2018) found that found that the seagrasses
Posidonia oceanica and Cymodocea nodosa also tended to accumulate
metals in either the leaves or the roots, like Z. capensis. The closely
related Z. marina has previously been found to employ this removal
strategy and has a potential as a bioindicator (Lee et al., 2019; Lyngby
and Brix, 1984; Williams et al., 1994).
Spartina maritima and S. tegetaria total biomass showed little signif­
icant seasonality, grasses (i.e. Z. capensis and S. maritima) contribute
significantly to the leaf litter in the estuary (Sousa et al., 2010). Since
S. maritima and S. tegetaria accumulate the majority of metals to the
roots, indicated by their high BCF (Fig. 4). This characteristic makes
these plants ideal in preventing mobile metals from re-entering the
sediment, and the rest of the food chain. This characteristic was prev­
alent even for Spartina densiflora growing in a relatively uncontaminated
Patagonian salt marsh, Argentina, displaying higher metal concentra­
tion in its belowground tissue (Idaszkin et al., 2014). Spartina maritima
and S. tegetaria, have been documented to be phytostabilisers, reducing
the mobility of heavy metals in the sediment. Duarte et al., 2010
concluded that Sarcocornia perennis was a much better phytostabiliser
species compared to S. maritima. Salicornia tegetaria, a closely related
species, also maintain more phytostabilisation than S. maritima. Root
senescence returns high amounts of metals to the sediment, and has been
found to occur for S. maritima, but its cycling rates between roots and
sediment occur slowly (Duarte et al., 2010). Our results indicate that a
decrease in root biomass of S. maritima between autumn and winter,
corroborating the results of previous studies. Maricle and Lee (2002)
showed that S. maritima promotes oxygenation of the roots with its
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Marine Pollution Bulletin 192 (2023) 115007
highly developed aerenchyma system, which may counteract metal
mobility in sediment together with root senescence. The relatively stable
root biomass of S. tegetaria suggests a slow root turnover rate, as found
for Sarcocornia perennis (a closely related, similar lifeform), promotes
phytostabilisation. Formation of root plaque in the rhizosediment of
these plants also assists their potential as phytostabilisers (Doyle and
Otte, 1997; Reboreda and Caçador, 2007).
This study showed that Z. capensis and S. tegetaria are valuable
standing stocks of metals in the Swartkops Estuary (Fig. 6). Similarly,
there were lower standing stocks for S. maritima compared to Halimoine
portulacoides, Sarcocornia fruticose and S. perennis reported by Caçador
et al. (2009). This confirms that S. tegetaria is a valuable sink for metals
in the Swartkops Estuary. Zostera capensis is a more valuable sink of
metals than previously known, both in the roots and the leaves. The
significantly lower standing stock found in S. maritima, and its well-
documented characteristic of being a hyperaccumulator (Curado et al.,
2014), indicate that the potential of uptake has not been exceeded in the
Swartkops Estuary.
Our results showed different total metal stocks for the studied spe­
cies, indicating autumn concentrations (Fe, Mn, Zn, Cr, Cu) in Z. capensis
far exceeded concentrations in S. tegetaria (except for Pb and Ni). It is an
increase in essential metals in Z. capensis that significantly exceeds the
stock in S. tegetaria. This stockpiling of metals come in the season with
the lowest total biomass values for Z. capensis. Internal concentrations of
essential metals can be higher than non-essential metals, since plants
have a greater capacity to take up these metals (Kabata-Pendias, 2011).
The spring growing season showed the lowest metal stock in S. tegetaria
and S. maritima coinciding with the highest leaf biomass for the season of
these plants. In the Swartkops Estuary, flowering for S. maritima was
observed in summer, while no flowering was observed for S. tegetaria.
Zostera capensis flowered in spring, but no indication of higher or lower
metal stocks were given, instead metals steadily decreased from the
colder seasons to the warmer seasons (autumn to summer), likely due to
growth dilution and physiological needs in warmer seasons. Hence, the
average standing stock in all three plants, displayed the same pattern as
that of the metal concentrations in sediments, i.e., Fe > Mn > Zn > Cr >
Pb > Ni > Cu. This indicated limited exclusion of certain metals at the
roots.
5. Conclusion
This study investigated the metal storage capabilities of three
wetland plant species, S. tegetaria, S. maritima, and Z. capensis in the
Swartkops Estuary, South Africa. Seasonal BCF and TF was unique to
each species, corroborating previous research indicating that metal
uptake cannot be compared between similar lifeforms or genera. Sali­
cornia tegetaria showed the highest potential as a phytostabiliser, due to
its low root turnover rate and high metal stock (μg m− 2). Although,
S. maritima is known for its high root accumulation, which was also
shown in this study, its lower comparable metal stock indicated a lower
potential for phytostabilisation compared to S. tegetaria. The seagrass,
Z. capensis, displayed capacity as a bioindicator in its roots (Cu, Ni, Cr),
but also accumulated metals in its leaves. Moreover, although
S. maritima is widely studied as it also occurs in parts of Europe, no
research in this field exists for the Southern African endemic, Z. capensis.
The ecological value of salt marsh and seagrass beds are evident in
heavily developed estuaries, but research needs to be conducted on all
species to establish each species’ worth.
CRediT authorship contribution statement
LH, JBA and GR conceptualized research, MN and LH wrote the first
draft, LH, MN collected field data, MN analysed data. LH, JBA, GR
provided inputs to data analysis and interpretation. All authors provided
editorial inputs to the paper.
M.A. Nel et al.
Funding
This work was funded by the National Research Foundation of South
Africa (Grant Number: 120406) awarded to Dr. L.R.D Human. Ms. M.A.
Nel was supported by a scholarship from the National Research Foun­
dation of South Africa (Grant Number: 116920).
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
Acknowledgements
This work was supported by the Shallow Marine and Coastal
Research Infrastructure (SMCRI) platform hosted by the South African
Environmental Observation Network Elwandle Node (SAEON). The
authors also wish to thank the Shallow Water Ecosystem research group
(funded by UID 84375DSI/NRF Research Chair) for assistance in the
laboratory and field.
References
Adams, J.B., 2020. Salt marsh at the tip of Africa: patterns, processes and changes in
response to climate change. Estuar. Coast. Shelf Sci. 237, 106650 https://doi.org/
10.1016/j.ecss.2020.106650.
Adams, J.B., Pretorius, L., Snow, G.C., 2019. Deterioration in the water quality of an
urbanised estuary with recommendations for improvement. Water SA 45, 86–96.
https://doi.org/10.4314/wsa.v45i1.10.
Adams, J., Hughes, D., James, N., 2021. Swartkops estuary: present ecological status and
future restoration scenarios. In: Institute for Coastal & Marine Research Report No.
48, p. 167.
Ali, H., Khan, E., Sajad, M.A., 2013. Phytoremediation of heavy metals-concepts and
applications. Chemosphere 91, 869–881. https://doi.org/10.1016/j.
chemosphere.2013.01.075.
Binning, K., Baird, D., 2001. Survey of heavy metals in the sediments of the Swartkops
River estuary, Port Elizabeth South Africa. Water SA 27, 461–466. https://doi.org/
10.4314/wsa.v27i4.4958.
Bonanno, G., Borg, J.A., 2018. Comparative analysis of trace element accumulation in
seagrasses Posidonia oceanica and Cymodocea nodosa: biomonitoring applications
and legislative issues. Mar. Pollut. Bull. 128, 24–31. https://doi.org/10.1016/j.
marpolbul.2018.01.013.
Bonanno, G., Vymazal, J., 2017. Compartmentalization of potentially hazardous
elements in macrophytes: insights into capacity and efficiency of accumulation.
J. Geochem. Explor. 181, 22–30. https://doi.org/10.1016/j.gexplo.2017.06.018.
Bonanno, G., Borg, J.A., Di Martino, V., 2017. Levels of heavy metals in wetland and
marine vascular plants and their biomonitoring potential: a comparative assessment.
Sci. Total Environ. 576, 796–806. https://doi.org/10.1016/j.scitotenv.2016.10.171.
Bonanno, G., Vymazal, J., Cirelli, G.L., 2018. Translocation, accumulation and
bioindication of trace elements in wetland plants. Sci. Total Environ. 631–632,
252–261. https://doi.org/10.1016/j.scitotenv.2018.03.039.
Bornman, T.G., Schmidt, J., Adams, J.B., Mfikili, A.N., Farre, R.E., Smit, A.J., 2016.
Relative Sea-level rise and the potential for subsidence of the swartkops estuary
intertidal salt marshes, South Africa. S. Afr. J. Bot. 107, 91–100. https://doi.org/
10.1016/j.sajb.2016.05.003.
Burke, D.J., Weis, J.S., Weis, P., 2000. Release of metals by the leaves of the salt marsh
grasses Spartina alterniflora and Phragmites australis. Estuar. Coast. Shelf Sci. 51
(2), 153–159.
Caçador, I., Vale, C., Catarino, F., 2000. Seasonal variation of zn, pb, cu and cd
concentrations in the root- sediment system of Spartina maritima and Halimione
portulacoides from tagus estuary salt marshes. Mar. Environ. Res. 49, 279–290.
https://doi.org/10.1016/S0141-1136(99)00077-X.
Caçador, I., Costa, A.L., Vale, C., 2004. Carbon storage in tagus salt marsh sediments.
Water Air Soil Pollut. Focus 4, 701–714. https://doi.org/10.1023/B:
WAFO.0000028388.84544.ce.
Caçador, I., Caetano, M., Duarte, B., Vale, C., 2009. Stock and losses of trace metals from
salt marsh plants. Mar. Environ. Res. 67, 75–82. https://doi.org/10.1016/j.
marenvres.2008.11.004.
Colloty, B.M., Adams, J.B., Bate, G.C., 2000. The use of a botanical importance rating to
assess changes in the flora of the swartkops estuary over time. Water SA 26,
171–180.
11
Marine Pollution Bulletin 192 (2023) 115007
Curado, G., Rubio-Casal, A.E., Figueroa, E., Castillo, J.M., 2014. Potential of Spartina
maritima in restored salt marshes for phytoremediation of metals in a highly
polluted estuary. Int. J. Phytorem. 16, 1209–1220. https://doi.org/10.1080/
15226514.2013.821451.
Deng, H., Ye, Z.H., Wong, M.H., 2004. Accumulation of lead, zinc, copper and cadmium
by 12 wetland plant species thriving in metal-contaminated sites in China. Environ.
Pollut. 132, 29–40. https://doi.org/10.1016/j.envpol.2004.03.030.
Di Leo, A., Annicchiarico, C., Cardellicchio, N., Spada, L., Giandomenico, S., 2013. Trace
metal distributions in Posidonia oceanica and sediments from Taranto gulf (Ionian
Sea, southern Italy). Mediterr. Mar. Sci. 14, 204. https://doi.org/10.12681/
mms.316.
Doyle, M., Otte, M., 1997. Organism-induced accumulation of iron, zinc and arsenic in
wetland soils. Environ. Pollut. 96, 1–11. https://doi.org/10.1016/S0269-7491(97)
00014-6.
Du Laing, G., Tack, F.M.G., Verloo, M.G., 2003. Performance of selected destruction
methods for the determination of heavy metals in reed plants (Phragmites australis).
Anal. Chim. Acta 497, 191–198. https://doi.org/10.1016/j.aca.2003.08.044.
Duarte, B., Caetano, M., Almeida, P.R., Vale, C., Caçador, I., 2010. Accumulation and
biological cycling of heavy metal in four salt marsh species, from tagus estuary
(Portugal). Environ. Pollut. 158, 1661–1668. https://doi.org/10.1016/j.
envpol.2009.12.004.
Duarte, B., Vaz, N., Valentim, J.M., Dias, J.M., Silva, H., Marques, J.C., Sleimi, N.,
Caçador, I., 2017. Revisiting the outwelling hypothesis: modelling salt marsh detrital
metal exports under extreme climatic events. Mar. Chem. 191, 24–33. https://doi.
org/10.1016/j.marchem.2016.12.002.
Feng, H., Qian, Y., Cochran, J.K., Zhu, Q., Hu, W., Yan, H., Li, L., Huang, X., Chu, Y.S.,
Liu, H., Yoo, S., Liu, C.J., 2017. Nanoscale measurement of trace element
distributions in Spartina alterniflora root tissue during dormancy. Sci. Rep. 7
https://doi.org/10.1038/srep40420.
Fourqurean, J.W., Johnson, B., Kauffman, J.B., Kennedy, H., Lovelock, C.E.,
Megonigal, J.P., Rahman, A., Saintilan, N., Simard, M., 2017. Coastal Blue Carbon.
Arlington, Virginia, USA.
Gedan, K.B., Silliman, B.R., Bertness, M.D., 2009. Centuries of human-driven change in
salt marsh ecosystems. Annu. Rev. Mar. Sci. 1, 117–141. https://doi.org/10.1146/
annurev.marine.010908.163930.
Greco, M., Sáez, C.A., Contreras, R.A., et al., 2019. Cadmium and/or copper excess
induce interdependent metal accumulation, DNA methylation, induction of metal
chelators and antioxidant defences in the seagrass Zostera marina. Chemosphere
224, 111–119. https://doi.org/10.1016/j.chemosphere.2019.02.123.
Guerinot, M.L., Ying, Yi, 1994. Iron: nutritious, noxious, and not readily available. Plant
Physiol. 104, 815–820. https://doi.org/10.1104/pp.104.3.815.
Helfield, J.M., Diamond, M.L., 1997. Use of constructed wetlands for urban stream
restoration: a critical analysis. Environ. Manag. 21, 329–341. https://doi.org/
10.1007/s002679900033.
Hoven, H.M., 1998. Eelgrass (Zostera marina L.) as a Sentinel Accumulator of Lead in
Portsmouth Harbor, New Hampshire - Maine doi:9907586.
Human, L., Feijão, E., de Carvalho, R.C., Caçador, I., Reis-Santos, P., Fonseca, V.,
Duarte, B., 2020. Mediterranean salt marsh sediment metal speciation and
bioavailability changes induced by the spreading of non-indigenous Spartina patens.
Estuar. Coast. Shelf Sci. 243, 1–9.
Idaszkin, Y.L., Bouza, P.J., Marinho, C.H., Gil, M.N., 2014. Trace metal concentrations in
Spartina densiflora and associated soil from a patagonian salt marsh. Mar. Pollut.
Bull. 89 (1–2), 444–450.
Kabata-Pendias, A., 2011. Trace Elements in Soils and Plants, Fourth ed. Taylor & Francis
Group. CRC Press, Boca Raton.
Lee, G., Suonan, Z., Kim, S.H., Hwang, D.W., Lee, K.S., 2019. Heavy metal accumulation
and phytoremediation potential by transplants of the seagrass Zostera marina in the
polluted bay systems. Mar. Pollut. Bull. 149 https://doi.org/10.1016/j.
marpolbul.2019.110509.
Li, C., Wang, H., Liao, X., Xiao, R., Liu, K., Bai, J., Li, B., He, Q., 2022. Heavy metal
pollution in coastal wetlands: a systematic review of studies globally over the past
three decades. J. Hazard. Mater. 424, 127312.
Lyngby, J.E., Brix, H., 1984. The uptake of heavy metals in eelgrass Zostera marina and
their effect on growth. Ecol. Bull. 36, 81–89.
Maricle, B.R., Lee, R.W., 2002. Aerenchyma development and oxygen transport in the
estuarine cordgrasses Spartina alterniflora and S. Anglica. Aquat. Bot. 74, 109–120.
https://doi.org/10.1016/S0304-3770(02)00051-7.
Matusiak, K., Skoczen, A., Setkowicz, Z., Kubala-Kukus, A., Stabrawa, I., Ciarach, M.,
Janeczko, K., Jung, A., Chwiej, J., 2017. The elemental changes occurring in the rat
liver after exposure to PEG-coated iron oxide nanoparticles: total reflection x-ray
fluorescence (TXRF) spectroscopy study. Nanotoxicology 11, 1225–1236. https://
doi.org/10.1080/17435390.2017.1408151.
McOwen, C.J., Weatherdon, L.V., Bochove, J.W., Sullican, E., Blyth, S., Zockler, C.,
Stanwell-Smith, D., Kingston, N., Martin, C.S., Spalding, M., Fletcher, S., 2017.
A global map of saltmarshes. Biodivers. Data J. 5.
Montero Alvarez, A., Estévez Alvarez, J.R., Padilla Alvarez, R., 2007. Heavy metal
analysis of rainwaters: a comparison of TXRF and ASV analytical capabilities.
J. Radioanal. Nucl. Chem. 273, 427–433. https://doi.org/10.1007/s10967-007-
6895-7.
Nel, L., Strydom, N.A., Bouwman, H., 2015. Preliminary assessment of contaminants in
the sediment and organisms of the swartkops estuary, South Africa. Mar. Pollut. Bull.
101, 878–885. https://doi.org/10.1016/j.marpolbul.2015.11.015.
Nel, M.A., Rubidge, G., Adams, J.B., Human, L.R.D., 2020. Rhizosediments of salicornia
tegetaria indicate metal contamination in the intertidal estuary zone. Front. Environ.
Sci. 8, 175.
M.A. Nel et al.
Phillips, D.P., Human, L.R.D., Adams, J.B., 2015. Wetland plants as indicators of heavy
metal contamination. Mar. Pollut. Bull. 92, 227–232. https://doi.org/10.1016/j.
marpolbul.2014.12.038.
Pretorius, L., 2015. Spatial and Temporal Variability in Water Quality Characteristics of
the Swartkops Estuary. Nelson Mandela Metropolitan University, South Africa. MSc
thesis.
Quan, W.M., Han, J.D., Shen, A.L., Ping, X.Y., Qian, P.L., Li, C.J., Shi, L.Y., Chen, Y.Q.,
2007. Uptake and distribution of N, P and heavy metals in three dominant salt marsh
macrophytes from Yangtze River estuary, China. Mar. Environ. Res. 64 (1), 21–37.
Ragsdale, H.L., Thorhaug, A., 1980. Trace metal cycling in the U.S. Coastal zone: a
synthesis. Am. J. Bot. 67, 1102–1112. https://doi.org/10.1002/j.1537-2197.1980.
tb07743.x.
Reboreda, R., Caçador, I., 2007. Halophyte vegetation influences in salt marsh retention
capacity for heavy metals. Environ. Pollut. 146, 147–154. https://doi.org/10.1016/
j.envpol.2006.05.035.
Sánchez-Quiles, D., Marbà, N., Tovar-Sánchez, A., 2017. Trace metal accumulation in
marine macrophytes: hotspots of coastal contamination worldwide. Sci. Total
Environ. 576, 520–527. https://doi.org/10.1016/j.scitotenv.2016.10.144.
Shilla, D., Pajala, G., Routh, J., Dario, M., Kristoffersson, P., 2019. Trophodynamics and
biomagnification of trace metals in aquatic food webs: the case of rufiji estuary in
Tanzania. Appl. Geochem. 100, 160–168. https://doi.org/10.1016/j.
apgeochem.2018.11.016.
Soda, S., Hamada, T., Yamaoka, Y., Ike, M., Nakazato, H., Saeki, Y., Kasamatsu, T.,
Sakurai, Y., 2012. Constructed wetlands for advanced treatment of wastewater with
a complex matrix from a metal-processing plant: bioconcentration and translocation
factors of various metals in Acorus gramineus and Cyperus alternifolius. Ecol. Eng.
39, 63–70. https://doi.org/10.1016/j.ecoleng.2011.11.014.
Sousa, A.I., Lillebø, A.I., Pardal, M.A., Caçador, I., 2010. The influence of Spartina
maritima on carbon retention capacity in salt marshes from warm-temperate
estuaries. Mar. Pollut. Bull. 61, 215–223. https://doi.org/10.1016/j.
marpolbul.2010.02.018.
Sulla, V., Zikhali, P., 2018. Overcoming Poverty and Inequality in South Africa: An
Assessment of Drivers, Constraints and Opportunities. Washington DC.
Turpie, J.K., Adams, J.B., Joubert, A., Harrison, T.D., Colloty, B.M., Maree, R.C.,
Whitfield, A.K., Wooldridge, T.H., Lamberth, S.J., Taljaard, S., Van Niekerk, L.,
12
Marine Pollution Bulletin 192 (2023) 115007
2002. Assessment of the conservation priority status of south african estuaries for use
in management and water allocation. Water SA 28, 191–206. https://doi.org/
10.4314/wsa.v28i2.4885.
Usman, A.R.A., Lee, S.S., Awad, Y.M., Lim, K.J., Yang, J.E., Ok, Y.S., 2012. Soil pollution
assessment and identification of hyperaccumulating plants in chromated copper
arsenate (CCA) contaminated sites, Korea. Chemosphere 87, 872–878. https://doi.
org/10.1016/j.chemosphere.2012.01.028.
Van Niekerk, L., Adams, J.B., Lamberth, S.J., MacKay, C.F., Taljaard, S., Turpie, J.K.,
Raimondo, D.C., 2019. South African National Biodiversity Assessment 2018:
Technical Report. Volume 3: Estuarine Realm. CSIR, Stellenbosch.
Vilhena, J.C.E., Amorim, A., Ribeiro, L., Duarte, B., Pombo, M., 2021. Baseline study of
trace element concentrations in sediments of the intertidal zone of amazonian
oceanic beaches. Front. Mar. Sci. 8 https://doi.org/10.3389/fmars.2021.671390.
Walsh, G.E., Grow, T.E., 1973. Composition of thulassia testttdinum and Ruppia
maritima. Q. J. Florida Acad. Sci. 35 (2), 97–108.
Wang, Z.L., Liu, C.Q., 2003. Distribution and partition behavior of heavy metals between
dissolved and acid-soluble fractions along a salinity gradient in the changjiang
estuary, eastern China. Chem. Geol. 202, 383–396. https://doi.org/10.1016/j.
chemgeo.2002.05.001.
Weis, J.S., Weis, P., 2004. Metal uptake, transport and release by wetland plants:
implications for phytoremediation and restoration. Environ. Int. 30, 685–700.
https://doi.org/10.1016/j.envint.2003.11.002.
Williams, T.P., Bubb, J.M., Lester, J.N., 1994. Metal accumulation within salt marsh
environments: a review. Mar. Pollut. Bull. 28, 277–290.
Windham, L., Weis, J.S., Weis, P., 2003. Uptake and distribution of metals in two
dominant salt marsh macrophytes, Spartina alterniflora (cordgrass) and Phragmites
australis (common reed). Estuar. Coast. Shelf Sci. 56, 63–72. https://doi.org/
10.1016/S0272-7714(02)00121-X.
Wiszniewska, A., Kamińska, I., Koźmińska, A., Hanus-Fajerska, E., 2018. Aspects of co-
tolerance towards salt and heavy metal stresses in halophytic plant species. In:
Hasanuzzaman, M., Fujita, M., Oku, H., et al. (Eds.), Plant Nutrients and Abiotic
Stress Tolerance. Springer Singapore, Singapore, pp. 477–498.
Yruela, I., 2009. Copper in plants: acquisition, transport and interactions. Funct. Plant
Biol. 36, 409–430. https://doi.org/10.1071/FP08288.