Water Air Soil Pollut (2011) 216:319328 DOI 10.

1007/s11270-010-0535-5

Accumulation and Distribution of Metals in Bolboschoenus maritimus (Cyperaceae), from a South African River
Likentso S. Shuping & Reinette G. Snyman & James P. Odendaal & Patrick A. Ndakidemi

Received: 4 March 2010 / Accepted: 29 June 2010 / Published online: 16 July 2010 # Springer Science+Business Media B.V. 2010

Abstract The Diep River is a major freshwater ecosystem in the Western Cape, South Africa. Although it is surrounded by many sources of metal pollution, the actual metal levels in this river system are unknown. Wetland plants are known to accumulate metals and may possibly be used as biomonitors of metal contamination in a river system. One such species, the sedge Bolboschoenus maritimus, is found in abundance along the banks of this river. The aim of this study was to investigate and monitor the degree of metal contamination in the water and sediments of the lower Diep River, as well as to study the seasonal accumulation and distribution of metals in B. maritimus, and the use thereof as biomonitor species. Two

sampling sites were used: one site above the wetland section of the river, receiving runoff mainly from agricultural lands (site 1), and one site close to the river mouth (site 2), exposed to several possible pollution sources. Water, sediment, and plant (root, leaf and stem) samples were collected seasonally for 1 year and analyzed for Al, Zn, Cu, and Fe. There was greater bioaccumulation of metals by plants at site 2, due to greater bioavailability of metals. B. maritimus was shown to be a root accumulator of metals. Seasonal fluctuations in root, stem, and leaf metal concentrations did not follow seasonal sediment concentration patterns. However, using B. maritimus as test species did provide valuable additional information to sediment and water analyses. More extensive research is needed to conclude whether this species is an effective biomonitor in the lower Diep River environment. Keywords Metals . Sedge . Root accumulator . Biomonitor

L. S. Shuping : J. P. Odendaal Department of Environmental and Occupational Studies, Faculty of Applied Sciences, Cape Peninsula University of Technology, PO Box 652, Cape Town 8000, South Africa R. G. Snyman (*) Department of Biodiversity and Conservation, Faculty of Applied Sciences, Cape Peninsula University of Technology, PO Box 652, Cape Town 8000, South Africa e-mail: snymanr@cput.ac.za P. A. Ndakidemi Department of Horticultural Sciences, Faculty of Applied Sciences, Cape Peninsula University of Technology, PO Box 652, Cape Town 8000, South Africa

1 Introduction Metals enter river systems through natural geochemical processes (Klavin et al. 2000) but concentrations are greatly increased anthropogenically, through, for example, effluent from wastewater treatment plants (Okonkwo and Mothiba 2005), industries, mining, urban developments, power stations, and agriculture (Botes and Van Staden 2005).

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Worldwide, researchers generally use sediments and water to assess pollution levels in aquatic ecosystems (e.g., Klavin et al. 2000; Grabowski et al. 2001; Adamiec and Helios-Rybicka 2002). This is also the case in South Africa (e.g., Binning and Baird 2001; Botes and Van Staden 2005; Okonkwo and Mothiba 2005). However, determining water and sediment metal loads alone does not provide any information on whether the metals are actually bioavailable to aquatic biota. Mortimer (1985) suggested that aquatic plants and animals must be incorporated as biomonitoring species in the evaluation of metal bioavailability and toxicity. Wetland plants are known to accumulate and store nutrients and metals in their tissues. This has been documented for species such as Phragmites australis (Bragato et al. 2006), Bolboschoenus maritimus (Almeida et al. 2006; Bragato et al. 2006; Madejn et al. 2006a), and Spartina alterniflora (Weis and Weis 2004; Weis et al. 2004) that can accumulate and store metals in their roots. B. maritimus is widely used in the reconstruction, creation and rehabilitation of wetlands (Kantrud 1996) and as test species in pollution-related studies in various countries around the world (Otte et al. 1991; Clevering 1995; Almeida et al. 2006; Madejn et al. 2006a). However, no such studies have been done in South Africa, even though the plant is commonly found on riverbanks around the Cape Peninsula and northwards (Trinder-Smith 2003). The Diep River is a major freshwater ecosystem in the Western Cape, South Africa, where B. maritimus is found in abundance. Before joining the Atlantic Ocean, the river forms a large wetland, of which part of it has been proclaimed a nature reserve. Upstream from the wetland, the river passes various possible sources of metal pollution, such as agricultural lands and a major landfill site. The wetland itself is surrounded by densely populated residential areas and industries, which are likely sources of pollution. A large sewage treatment plant also discharges into the lower part of the river. Despite all these possible pollution sources, no comprehensive studies have been done on metal contamination of this river. Also, the use of a biomonitor species to determine the degree of metal bioaccumulation has not been investigated. According to Madejn et al. (2006b), biomonitors in a soil environment are species that indicate soil quality and that reveal information on soil quality that is difficult to

measure using direct soil analyses. For example, they may confirm the availability of trace elements in the soil (Madejn et al. 2004). The latter authors suggested that plants that are trace element accumulators could be used as biomonitors. Based on the successful use of B. maritimus as a test species (although not a biomonitor) in other studies, particularly also in terms of its ability to accumulate metals (Otte et al. 1991; Clevering and Hundscheid 1998; Bragato et al. 2006; Madejn et al. 2006a), it was chosen as a potential biomonitor species for the present study. The objectives of this study were therefore to investigate and monitor the degree of metal contamination in the water and sediments of the lower Diep River, as well as to study the accumulation and distribution of metals in the sedge B. maritimus, and the use thereof as a biomonitor species.

2 Materials and Methods 2.1 Sampling Sites Two sites were selected in the lower reaches of the river. No reference site in the upper reaches could be used, since that part of the river has been dry for several years. Site 1 This site was located 6 km upstream of the wetland reserve. This site receives runoff mainly from agricultural lands. Other possible metal sources include municipal dumpsites. Site 2 This site was located downstream of the wetland, 2 km from the river mouth. Above this site are several possible pollution sources, including a large industrial and residential area and a sewage treatment plant.

2.2 Sampling Procedure Sediment, water and plant samples were collected seasonally, over a 1-year period, commencing with summer. Plants were completely uprooted and separated into roots, leaves and stems in the laboratory. Sediment samples were taken from the root zone, to a maximum depth of 10 cm. A water temperature, pH,

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and electrical conductivity reading were also taken at each site, during each sampling occasion. All water, sediment, and plant samples used for metal analysis were frozen at 18C until preparation for metal analysis could be done. Sediment samples used for characterization were not frozen. 2.3 Sediment Characterization One sample of top sediment from each site was used for sediment characterization. The analysis procedure followed was according to the method described by Folk (1954). Samples were separated into their different fractions, namely sand, silt, and clay, and the percentage of each determined. 2.4 Metal Analysis After thawing, 10 ml of each water sample was poured into a test tube. The plant specimens and the sediment samples were oven dried for 48 h at 60C to obtain the dry weight. Sediment was sieved to remove large particles and plant specimens were ground using a pestle and mortar. A subsample (1 g) of each dried sample was placed into a test tube for acid digestion. Acid digestion of all samples was done according to the method used by Odendaal and Reinecke (1999). Ten milliliters of 55% nitric acid were added to samples and a 10-ml blank was prepared. Samples were heated in a dry block heater in a fume cabinet, at 40C for 1 h. The temperature was then increased to 120C and maintained for 3 h. After acid digestion, samples were left to cool, and then diluted with distilled water to obtain a 20 ml sample. Finally, samples were filtered using 0.4 or 0.6 mm Whatman filter paper and 0.45 m cellulose nitrate membrane filter paper, a needle and a syringe, after which they were stored in a refrigerator. Metal concentrations were determined using an inductively coupled plasmaatomic emission spectrophotometer (ICP-AES). All sediment and plant metal concentrations were expressed as mg/kg dry mass, and all water metal concentrations as mg/l. The metals that were originally analyzed for were chromium, iron, cobalt, copper, aluminum, zinc, cadmium, nickel, and lead. Aluminum, iron, copper, and zinc were selected for this study, since these were actually detected in the river water. The detection limit of the four selected metals on the ICP-AES was 0.00001 ppm.

2.5 Statistical Analysis KruskalWallis one-way analysis of variance on ranks was carried out to compare the concentrations of metals at different sites, over the duration of the sampling period, in water, sediment and plant samples. The StudentNewmanKeuls method was used to do pairwise multiple comparisons. Statistical analysis was done using Sigmastat 3.1 software package.

3 Results 3.1 Water pH, Temperature, and Electrical Conductivity Mean (SD) water pH, temperature and electrical conductivity measured at the two sites, are shown in Table 1. Since only one reading was taken per sampling occasion, a mean for the entire sampling period was calculated per site. The electrical conductivity measured for site 2 was significantly higher (p< 0.05) than for site 1. Water pH and temperature did not differ significantly between the two sites. 3.2 Metal Concentrations in Sediments and Water Mean (SD) metal concentrations (mg/kg) in sediments and water, per sampling occasion, measured for sites 1 and 2, are shown in Table 2. Concentrations of aluminum and iron in the sediments were significantly higher (p<0.05) at site 1 for all four seasons. Copper and zinc concentrations in the sediments were significantly higher (p<0.05) at site 2, for all four seasons and three of the four seasons respectively (zinc was not detected in sediment from site 1 during
Table 1 Mean (SD) water pH, temperature (C) and electrical conductivity (mS/m), for the entire sampling period, measured at sites 1 and 2 in the lower Diep River Physico-chemical parameter pH Temperature (C) Electrical conductivity (mS/m) Site 1 7.450.31a 19.453.28a 270.5105.7
a

Site 2 7.320.9a 20.784.26a 527.5197.24b

N=4. Letters in superscript indicate significant differences (p<0.05) between sites

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summer). In the case of water, there were no significant differences (p>0.05) in metal concentrations between the two sites. 3.3 Sediment Characterization Sediment characterization showed that sediment at site 1 is slightly gravelly sandy mud (gsM), with a 19.78% clay fraction, 49.85% silt fraction and 29.35% sand fraction. Sediment at site 2 is gravelly sandy (gS), with a 0.44% clay fraction, 5.63% silt fraction and 79.41% sand fraction. 3.4 Metal Concentrations in Plant Roots, Leaves, and Stems Mean (SD) metal concentrations in plant roots, stems and leaves, per sampling occasion (season), measured for sites 1 and 2, are shown in Fig. 1ah. Overwhelmingly, metal concentrations were significantly higher (p<0.05) in the roots than in the aboveground parts, at both sites. Leaves and stems often did not differ significantly but where differences were found, metal concentrations were mostly higher in leaves than in stems. Where differences were found between sites, metal concentrations were mostly significantly higher (p<0.05) in plants from site 2. Varying results were found when seasons were compared and no clear seasonal patterns could be detected. Copper and zinc showed particularly high peaks in roots during winter (sampling occasion 3) and spring (sampling occasion 4) at site 2 (Fig. 1f and h).

4 Discussion and Conclusions Metal concentrations in water samples were compared with the South African Water Quality Guidelines for Aquatic Ecosystems (DWAF 1996). The Target Water Quality Range (TWQR) for aquatic ecosystems for aluminum is 0.01 mg/l, for copper is 0.0014 mg/l, and for zinc is 0.002 mg/l. Guideline values for iron are not available. Aluminum and zinc concentrations were almost always higher than the guideline concentrations at both sites. Copper was often below detectable levels but, in summer, concentrations above the TWQR were measured at both sites. When comparing water metal concentrations between the sites and between sampling occasions

(seasons), there seems to be no clear patterns or trends. The metals that were detected are all either micronutrients (zinc, copper, and iron) or metals contained in clay minerals (aluminum and iron; Ashman and Puri 2002). The small fluctuations in concentrations between sampling occasions and minor differences between sites were probably mainly due to natural metal fluctuations as a result of biological processes such as decomposition (Phillips 1977; Luoma et al. 1982) and ion uptake and excretion by plants (Sawidis and Voulgaropoulos 1986; Ho 1990) and animals (Hare 1992). Other possible reasons could be occasional suspension of sediment into the water, different physico-chemical forms of sediment-bound metals (Luoma and Bryan 1981) or ion exchange between the clay minerals in the sediment and the water (Warren and Rudolph 1997). Sediment aluminum and iron concentrations were significantly higher at site 1 and zinc and copper significantly lower (Table 2). Aluminum and iron are metals that are naturally included in the clay mineral structure (Ashman and Puri 2002). The higher clay content (montmorillonite) found in the sediment at site 1, and therefore also higher contents of Al and Fe oxides, is thus most probably responsible for the elevated concentrations of these two metals. Pollution is less likely to have contributed at this site, since no major pollution sources for these metals exist above this site. However, the presence of aluminum and iron at site 2, as well as the significantly higher zinc and copper concentrations here, show that input from the sewerage works and industries above this site are contributing to metal pollution in this section of the river. When the two sites were compared in terms of metals in the various plant components, it was often found that there were no significant differences. However, wherever significant differences were found, plants from site 2 overwhelmingly had the highest concentrations of all metals tested for. There was thus greater bioaccumulation by the plants from site 2, even of the metals occurring in lower concentrations at this site (Al and Fe). The most likely explanation for this is an increased bioavailability of these metals at this site. Metal bioavailability is influenced by various factors, such as pH, temperature, redox potential, chemical speciation, seasonal changes, sediment type, salinity, and organic matter (Otte et al. 1993; Du

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Table 2 Mean (SD) aluminum, iron, copper and zinc concentrations measured in sediments (mg/kg dry mass) and water (mg/l) of the lower Diep River, over four seasons (summer to spring) Sampling occasions (seasons) Metal Sediment Site 1 1: Summer 2. Autumn 3: Winter 4: Spring 1: Summer 2. Autumn 3: Winter 4: Spring 1: Summer 2. Autumn 3: Winter 4: Spring 1: Summer 2. Autumn 3: Winter 4: Spring N.D. not detected N=5. Letters in superscript indicate significant differences (p<0.05) between sites Zn Cu Fe Al 13,271.42501.05a 10,456.54238.24
a

Water Site 2 2,837.31377.64b 6,455.7275.53

a b

Site 1 1.470.18a 0.250.08 1.831.2a 0.130.1

a a a a a

Site 2 2.280.41a 1.430.81a 1.600.69a 0.270.06a 1.910.98a 20.11.57b 1.480.53a 0.110.1a

a

12,455.33681.58a 13,702.361,389.47 4,133.441,443.4

a a a

7,108.64966.31b 2,756.9143.45 2,960.13364.94 10,608.05369.59 7,639.92828.0 12.830.98

b b b b b b

2.631.28 0.620.36 1.880.88

18,181.26238.09 17,951.47793.07 4.181.2

a a

16,150.37682.04a 4.921.13

3,072.31131.04b 9.611.09

0.240.17a 0.0020.006 N.D. N.D. N.D. 0.210.02

a a a a

0.040.01a N.D. N.D. N.D. 0.120.08a 1.700.04a 0.0320.027a 0.020.01a

12.420.83a 10.00.91a N.D. 83.3940.28 28.61.43

a a a

24.31.74b 15.591.45b 11.573.04 589.77159.67 82.5215.06 32.611.12

b b b

1.070.03 0.020.01

29.531.77

0.0080.007

Laing et al. 2007). Bioavailability in this study may have been influenced by several of these factors however, since water pH and temperature did not differ significantly between the two sites (Table 1), these factors are unlikely to have played a major role. The significantly higher electrical conductivity measured at site 2, due to pollutants and seawater influx at high tides (Grindley and Dudley 1988), indicate that salinity most likely increased metal bioavailability substantially here. An increase in salinity is known to increase metal bioavailability to plants (Fitzgerald et al. 2003; Stevens et al. 2003). Otte et al. (1991) also found such an increase in metal concentrations in Aster tripolium under saline conditions. Apart from salinity, sediment type would also have strongly influenced bioavailability. The sediment at site 1 has a high clay content, which could lead to metals binding to sediment and not being readily available for plants. Clay compounds have demonstrated binding potential and reduced metal solubility in soil solution (Lothenbach et al. 1999). At site 2, on the other hand, where the sediment lacks clay, metals may as a result be more available to plants.

Plant components (roots, leaves, and stems) were compared with one another, per site, to determine the distribution of metals in B. maritimus (Fig. 1ah). In general, metal concentrations were significantly higher in roots than in any other plant component. Higher metal content in the roots is to be expected, according to Almeida et al. (2006), since the root system is the main uptake pathway of metals from the sediment. However, numerous authors, e.g., Otte et al. 1991; Peverly et al. 1995; Weis and Weis 2004; Almeida et al. 2006; Madejn et al. 2006a) found that various wetland plants actually accumulate and immobilize certain metals in their root tissues, thus limiting distribution to aboveground parts. Of these authors, Otte et al. (1991), Almeida et al. (2006) and Madejn et al. (2006a), in particular, all concluded that B. maritimus can be considered a root accumulator of metals. The results of the present study are in agreement with the results of these latter authors and present a possibility of using this plant in rhizofiltration. Bioaccumulation of metals in belowground tissues is a strategy that plants use to restrict distribution of metals to aboveground tissues, to avoid damage of

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Roots Leaves Stems

A (Site1)
14000

B (Site2)
Concentration (mg/kg)
14000 12000 10000 8000 6000 4000 2000 0
1

Roots

Leaves

Stems

Concentration (mg/kg)

12000 10000 8000 6000 4000 2000 0

1 2 3 4

Sampling occasions

Sampling occasions

C (Site 1)
18000

Roots

Leaves

Stems

D (Site2)
18000

Roots

Leaves

Stems

Concentration (mg/kg)

Concentration (mg/kg)

16000 14000 12000 10000 8000 6000 4000 2000 0

1 2 3 4

16000 14000 12000 10000 8000 6000 4000 2000 0

1 2 3 4

Sampling occasions

Sampling occasions

E (Site1)
Concentration (mg/kg)
60 50 40 30 20 10 0

Roots

Leaves

Stems

F (Site 2)
60

Roots

Leaves

Stems

Concentration (mg/kg)
1 2 3 4

50 40 30 20 10 0

Sampling occasions

Sampling occasions

G (Site 1)
Concentration (mg/kg)
180 160 140 120 100 80 60 40 20 0
1

Roots

Leaves

Stems

H (Site2)
180

Roots

Leaves

Stems

Concentration (mg/kg)
2 3 4

160 140 120 100 80 60 40 20 0 1 2 3 4

Sampling occasions

Sampling occasions

Fig. 1 Mean (SD) aluminum (a and b), iron (c and d), copper (e and f) and zinc (g and h) concentrations (mg/kg, dry mass) measured in roots, leaves, and stems of B. maritimus collected

from two sites in the Diep River, over four seasons (summer to spring). N=5

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their photosynthetic tissues (Deng et al. 2004; Bragato et al. 2006). Metals are stored in the inner root tissues, within cells of the stele, or within cell walls and vacuoles (Weis and Weis 2004). Metals may also be immobilized through the production of metallothioneins (low molecular weight proteins which bind metals) and phytochelatins (synthesized enzymatically from glutathione in response to many metals, they chelate and bind metals; Rauser 1990; Memon et al. 2001). It must be noted that metal plaques on root surfaces may affect concentrations measured in roots. Plaques are composed mostly of iron hydroxides and other metals such as manganese that are mobilized and precipitated on the root surface (Weis and Weis 2004). Apart from the fact that they could give a false impression about metal concentrations contained inside roots, they could also increase or decrease the uptake of metals by plants. Iron plaques are known to adsorb large amounts of cations and on roots may, for example, form a barrier to the uptake of cationic nutrients and metals and may immobilize metals (Salomons and Frstner 1984; Taggart et al. 2005, 2009). It is possible that metal plaques may have, to some extent, influenced metal concentrations measured for the roots in the present study. However, the authors assume that the largest part of the metals measured, were in fact contained inside the roots. This assumption is based on a large body of literature on B. maritimus and root accumulation (as discussed previously). Metals were not only detected in the roots of B. maritimus in the present study, but were also detected in the stems and leaves. This is easily explained for Cu, Zn, and Fe as they are essential plant micronutrients (Madejn et al. 2006a) and therefore need to be distributed to all parts of the plant. They may even be found in higher concentrations aboveground, as shown for Zn and Cu in Spartina maritima and Halimione portulacoides (Cacador et al. 2000). The aluminum measured in the leaves and stems had probably been translocated from the roots. This translocation may have occurred due to one or more of the following factors: (1) exceeding of the root storage capacity: excess metals are stored in the roots and if the storage capacity is exceeded, the metals are translocated to the aboveground tissues regardless of whether they are essential or non-essential (Otte et al. 1991). (2) Senescence of aboveground tissues, espe-

cially leaves: Bragato et al. (2006) suggested that when photosynthetic activities are reduced due to aging of leaves, plants translocate metals to the aging parts as a mechanism to reduce metal burden. Luque et al. (1999) also found that old leaves had higher concentrations of various metals than young leaves. In the present study, older leaves were part of the whole plants sampled for metal analysis. This may have influenced the results. (3) Increased transpiration by leaves, leading to increased metal uptake in the process: Otte et al. (1991) found increased concentrations of metals in A. tripolium tissues under saline conditions, which they suggested may have been related to higher water uptake due to increased transpiration, leading to higher flux of metals into the entire plant. In the present study, electrical conductivity readings were particularly high at site 2, thus possibly increasing transpiration, water and metal uptake rates, and metal distribution rates in the plants. (4) Other metals and elements present in the plant: these may increase or decrease the accumulation of metals by plants, as well as influence the distribution, sequestration and immobilization of metals in specific plants parts. Weis et al. (2004) found that Cu distribution was affected by the presence of Zn or Pb in P. australis. Fritioff and Greger (2006) found a decrease of Cd in the roots and an increase of Cu in leaves of Potamogeton natans, in the presence of other metals in these tissues. A clear relationship was also found between Al and Ca in the foliage of Picea rubens (Borer et al. 2004). Seasonal variations in metal concentrations in B. maritimus roots were observed, but these did not follow similar patterns between the different metals or between the two sites. These variations also did not correspond with fluctuations in sediment concentrations (Fig. 1ah). Seasonal fluctuations in root metal concentrations are influenced by many factors, notably bioavailability (Larsen and Schierup 1981), which in turn is affected by, amongst others, pH and temperature, as well as physiological factors of the plant itself (Otte et al. 1993). The fact that fluctuations in roots of B. maritimus mostly did not correspond with fluctuations in sediment concentrations clearly indicates the important influences of the above-mentioned factors on metal uptake in this plant. The high concentration peaks found for Cu and Zn in winter (sampling occasion 3) and spring (sampling occasion 4) at site 2, were probably also

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Water Air Soil Pollut (2011) 216:319328 Almeida, C. M. R., Mucha, A. P., & Vasconcelos, M. T. S. D. (2006). Comparison of the role of the sea club-rush Scirpus maritimus and the sea rush Juncus maritimus in terms of concentration, speciation and bioaccumulation of metals in the estuarine sediment. Environmental Pollution, 142, 151159. Ashman, M. R., & Puri, G. (2002). Essential soil science: A clear and concise introduction to soil science. Malden: Blackwell. Binning, K., & Baird, D. (2001). Survey of heavy metals in the sediments of the Swartkops River Estuary, Port Elizabeth South Africa. Water South Africa, 27(4), 461466. Borer, C. H., Schaberg, P. G., DeHayes, D. H., & Hawley, G. J. (2004). Accretion, partitioning and sequestration of calcium and aluminum in red spruce foliage: implications for tree health. Tree Physiology, 24, 929939. Botes, P. J., & Van Staden, J. F. (2005). Investigation of trace element mobility in river sediments using ICP-OES. Water South Africa, 31(2), 183191. Bragato, C., Brix, H., & Malagoli, M. (2006). Accumulation of nutrients and heavy metals in Phragmites australis (Cav.) Trin. Ex. Steudel and Bolboschoenus maritimus (L.) Palla in a constructed wetland of the Venice lagoon watershed. Environmental Pollution, 144(3), 967975. Cacador, 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. Marine Environmental Research, 49, 279290. Clevering, O. A. (1995). Germination and seedling emergence of Scirpus lacustris L. and Scirpus maritimus L. with special reference to the restoration of wetlands. Aquatic Botany, 50, 6377. Clevering, O. A., & Hundscheid, M. P. J. (1998). Plastic and non-plastic variation in growth of newly established clones of Scirpus (Bolboschoenus) maritimus L. grown at different water depths. Aquatic Botany, 62, 117. 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. Environmental Pollution, 132, 2940. Du Laing, G., Vanthuyne, D., Tack, F. M. G., & Verloo, M. G. (2007). Factors affecting metal mobility and bioavailability in the superficial intertidal sediment layer of the Scheldt estuary. Aquatic Ecosystem Health & Management, 10, 3340. DWAF. (1996). South African water quality guidelines, vol. 7: aquatic ecosystems (1st ed.). Pretoria: Department of Water Affairs and Forestry. Fitzgerald, E. J., Caffrey, J. M., Nesaratnam, S. T., & McLoughlin, P. (2003). Copper and lead concentrations in salt marsh plants on the Suir Estuary, Ireland. Environmental Pollution, 123, 6774. Folk, R. L. (1954). Distinction between grain size and mineral composition in sedimentary rock nomenclature. Journal of Geology, 62, 345351. Fritioff, A., & Greger, M. (2006). Uptake and distribution of Zn, Cu, Cd, and Pb in an aquatic plant Potamogeton natans. Chemosphere, 63, 220227.

caused by a combination of the above-mentioned factors. It is therefore important to establish the mechanisms used by B. maritimus to take up, accumulate, and transport metals in their organs, under different conditions. In conclusion, it is clear that the present study can serve as a foundation for future studies in this river. The lower Diep River is definitely contaminated with metals and may pose a threat to this ecosystem, as a result of their availability to aquatic organisms. Therefore, further monitoring should be undertaken on metal contamination and toxicity in this river, particularly since the river runs through an area where there is continuous development. Finally, there is some doubt as to whether B. maritimus can be effectively used as biomonitor species in an environment such as the lower Diep River, particularly since root concentrations mostly did not indicate the actual level of contamination in the environment (sediment), or temporal changes in contamination levels. On the other hand, B. maritimus did provide additional information that soil analyses alone would not have provided, namely the bioavailability of the metals in the sediment and water. According to Madejn et al. (2006b) and Mertens et al. (2006), such additional information is necessary for a species to qualify as a biomonitor species. However, Mertens et al. (2006) further stated that the information provided by a biomonitor species should ideally also include information on plant or ecosystem functioning. The results of the present study thus warrant further investigation in this regard, for example with the use of biomarkers and other toxicological endpoints.
Acknowledgments We thank the National Research Foundation and the Cape Peninsula University of Technology for financial assistance. We also thank J. Rogers of the Geology Department, University of Cape Town, for assistance with the sediment characterization.

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