Aquatic Botany, 29 (1988) 303-317 303

Elsevier Science Publishers B.V., Amsterdam - - Printed in The Netherlands

RHIZOSPHERE OXYGENATION BY TYPHA DOMINGENSIS

PERS. IN MINIATURE ARTIFICIAL WETLAND FILTERS USED
FOR METAL REMOVAL FROM WASTEWATERS

JANET S. DUNBABIN, JAN POKORNY 1 and KATHLEEN H. BOWMER 2

CSIRO Centre for Irrigation and Freshwater Research, Griffith, NSW 2680 (Australia)
(Accepted for publication 24 August 1987)

ABSTRACT

Dunbabin, J.S., Pokorn~, J. and Bowmer, K.H., 1988. Rhizosphere oxygenation by Typha dom-
ingensis Pers. in miniature artificial wetland filters used for metal removal from wastewaters.
Aquat. Bot., 29: 303-317.

Miniature artificial wetland filters were used to treat a metal-enriched synthetic effluent by
upward percolation through a gravel substratum. The systems were either planted with Typha
domingensis Pets. or unplanted, and were with or without lactic acid or ground leaves as carbon
amendments to enhance biological oxygen demand. Oxygen concentration, pH, redox potential
and metal retention were higher in planted systems than in systems without plants. Respiration
was increased by carbon amendments in all filters. Oxygen concentration in unplanted filters
declined, but rhizosphere aeration maintained oxygenated conditions in all planted filters. The
results demonstrate that available carbon supply improved heavy metal removal in the artificial
wetland filters and affected the rate of rhizosphere oxygenation. Rhizosphere oxygenation, cal-
culated from the sum of respiration and net oxygenation observed in static assay, was of the order
of at least 80-300 Hg 11 h -1, or 210-750 Hg h-1 filter-1.

INTRODUCTION

Emergent wetland plants growing in anoxic sediments are adapted to cope

with anaerobic conditions. Rhizosphere oxygenation is considered essential for
active root functions, and also enables the plants to counteract the effects of
soluble phytotoxins, including sulphides and metals, which may be present at
high concentrations in anoxic substrata (Armstrong, 1971; Gambrell and Pa-
trick, 1978).
Anaerobic metabolism may be a short-term adaptation to flooding (John
and Greenway, 1976) or to survival in extremely reducing environments

1Present address: Czechoslovak Academy of Sciences, Institute of Botany, Department of Hydro-

botany, Dukelska 145, 37982 T~ebofi, Czechoslovakia.
2Author to whom requests for reprints should be addressed.

nqn4 _qg~n/~ l~nq ~N ~ I ClRR l~,l~vi~r Rcience Publishers B.V.

304

( Mendelssohn et al., 1981 ) but anatomical adaptations, to allow air supply to

underground organs and roots, are important for survival and growth in the
longer term. Air diffuses from the leaves through aerenchymatous tissue to
submerged parts of the plant ( Conway, 1937; van Raalte, 1941; Teal and Kan-
wisher, 1966; Armstrong, 1978; Sale and Wetzel, 1983).
Aerobic conditions are required for the nitrification of ammoniacal nitrogen
compounds prior to their gaseous loss by denitrification (Thursby, 1984 ), and
for the oxidation of iron and manganese to hydrous oxides which have a high
surface area and adsorptive capacity for phosphorus and metals (Patrick and
Gambrell, 1976). Therefore, rhizosphere oxygenation may be an important
influence on the behaviour of natural and artificial wetlands. The latter have
potential for low maintenance, low-cost treatment of rural, urban and indus-
trial wastewaters for removal of nutrients and heavy metals (Finlayson and
Chick, 1983; Gersberg et al., 1985a). The investigations reported here are part
of a larger study to assess the suitability of artificial wetlands for treating see-
pages from mine tailings or water impoundments on mine sites after closure.
The extent of oxygenation will depend on several factors, including the ox-
ygen demand of biological and chemical processes in the substratum and the
leakiness of the roots for oxygen, which varies with plant species ( Armstrong,
1964). Evidence for oxygen diffusion outwards from the roots is derived from
observations of the precipitation of oxidised ferric iron and changes in redox
conditions close to root surfaces (Wium-Andersen and Andersen, 1970; Tes-
senow and Baynes, 1978; Chen et al., 1980; Taylor et al., 1984) and by direct
measurement using cylindrical platinum electrodes surrounding the roots
(Armstrong, 1971; Armstrong and Wright, 1975 ). However, there is very little
information on the overall effect of plant-induced oxygenation on wetland sub-
strata, or on their capacity to remove or inactivate metals.
Part of the metal entering an artificial wetland filter is retained in the sub-
stratum as a result of trapping of particulate matter, and precipitation and
adsorption reactions, part is taken up by plants and the remainder flows out
of the system (Banus et al., 1975, Lindau and Hossner, 1982; Gersberg et al.,
1985b ). The partitioning of metals between these phases is determined by the
pH and redox of the artificial filter system and by the presence of chelating
agents released during the decomposition of organic matter (Gambrell et al.,
1977). When high rates of respiration release C02, the pH of the system is
reduced, increasing the solubility and toxicity of the metals. Rhizosphere ox-
ygenation is required for the plants to counteract toxic effects by precipitating
the metals close to the root surfaces. Therefore, uptake of metals by plants,
organic matter decomposition, oxygenation, redox, pH and retention of metals
in the system are interrelated processes (Taylor et al., 1984).
In this experiment we used a series of miniature artificial wetland filters, on
which different treatments were imposed, in an attempt to quantify the role of
several of these factors. Each filter system consisted of a bed of gravel with or
 305

Fig. 1. Unplanted wetland filter dimensions showing (A) gravel surface, (B) water level, (C)
filler tube with flanged base to spread the influent, (D) reservoir with restricted inlet to reduce
turbulence on addition of influent, (E) position of oxygen probe during in situ measurements,
(F) outlet tube, used for collection of samples, (G) mesh bag containing ground leaves (where
supplement of T. dorningensis used) and (H) redox probe.

without Typha domingensis Pers. A synthetic effluent, enriched with metals,

was percolated upwards through the gravel substratum. The objectives were
(1) to quantify the net oxygen flux resulting from the opposing effects of rhi-
zosphere oxygenation and respiration in the substratum and ( 2 ) to investigate
the effects of oxygenation on the retention of copper (Cu), zinc (Zn) and
cadmium (Cd) in the wetland filters.
Daily measurements of the metal flowing out of each system were compared
with the load added to determine the retention of metals by artificial wetlands.
Two contrasting types of organic carbon source were added to stimulate mi-
crobial respiration: lactic acid was used as a readily available source and ground
Typha leaves were used as a natural amendment.

MATERIALS AND METHODS

Experimental wetlands and operation

Each experimental system (Fig. 1 ) consisted of a plastic bucket (101), either

unplanted, or containing T. domingensis growing in a gravel substratum (di-
ameter 25-75 mm). The plants were grown from seed collected at a smelting
works, and three young plants were planted in each bucket. The experiment
was conducted in a glasshouse.
To obtain a uniform stand of healthy plants, daily additions of nutrient so-
lution were made for 4 weeks before introducing metals to make a synthetic
wastewater (Appendix I ). Each morning a basal addition of cooled wastewater
(0.5 l, equivalent to about one sixth of the pore volume) was added through
the filler tube ( Fig. 1 ). In preliminary checks, dye dilution studies showed that
the density gradient produced by cooling this influent by 2-3 °C prevented
mixing until temperature equilibrium was reached at least 1 h after addition
(P.R. Breen, personal communication, 1985). Therefore, the samples dis-
306

placed upwards and collected through the three surface outlets were not mixed
with the recent influent. The systems were kept saturated throughout the ex-
periment. Evapotranspiration losses were replaced each evening by adding
metal-free nutrient solution from the surface. Dye measurements showed that
these were completely mixed with the bulk solution before sampling in the
morning. This design ensured that the redox status of the wetlands was mini-
mally disturbed, and that all the wastewater passed through the root zone with
a theoretical residence time within the filter system of 6 days. ( The theoretical
pore volume was determined by displacement of pore space in an unplanted
system. In planted systems, root growth reduced the pore volume to about
2.2-2.5 l, so that retention times were reduced from 6 days to between 4.4 and
5.0 days. )
The experiment was a three-way factorial design in which treatments were
as follows: planted or unplanted, with or without organic carbon as ground
leaves and with or without organic carbon as lactic acid. Each treatment was
replicated six times.

Organic carbon

Ground leaves of T. domingensis, equivalent to about 0.6% (w/w) of the

substratum, were placed in a fine mesh bag below the filler tube (Fig. 1 ). This
design allowed recovery at the end of the experiment, and analysis of metals
retained by the leaves. The lactic acid amendment, equivalent to a carbon con-
centration of 25 mg 11 in the wastewater, was added directly to the synthetic
effluent for daily addition. The available carbon load per wetland was about
29 and 1.225 g for the dry leaves and lactic acid, respectively, assuming 100%
availability of lactic acid and 45% availability of leaves (Westlake, 1963 ).

Metal addition and analysis

Addition of metals to the artificial filters began 4 weeks after planting. A

nutrient solution, as described in Appendix I, was enriched with metals for use
as a synthetic wastewater. Concentrations of Zn, Pb, Cu and Cd were 2.5, 1.0,
0.5 and 0.5 mg 11, respectively. The metals were added as ZnSO4.7H20,
Pb (NO3)2, CuSO4.5H20 and 3CdSO4.8H20. The initial pH was adjusted to
5.3 using sulphuric acid or potassium hydroxide, and conductivity was 700-800
~/S c m -1.
Effluent wastewater samples were collected each morning as daily additions
were made, and pH was recorded immediately. Subsamples were preserved in
concentrated nitric acid and refrigerated before metal analysis by flame atomic
absorption spectrophotometry.
Plants were harvested at the conclusion of the experiment, 18 weeks after
planting and 14 weeks after 'flow-through' operation began. The plants were
dried at 65 ° C, ground, digested with a mixture of nitric and perchloric acids
 307

(ratio 9:1 ), and analysed as before. Ground leaves of T. domingensis, used as

an organic carbon supplement, were analysed similarly both before and after
exposure to metal-enriched wastewater.

Redox

Redox potentials were measured at weekly intervals using five platinum

electrodes per filter system. The electrodes were permanently inserted into the
gravel substratum, with sensor tips at a depth of 0.10-0.15 m. A standard ref-
erence calomel half-cell filled with saturated potassium chloride solution was
used, and the reference potential ( 242 mV ) was added to the measured value.
All filter systems were used from one randomly selected block in the three-way
factorial design, making a total of eight sets of observations. Measurements
continued until 15 weeks after planting.

Oxygen measurements

Detailed measurements of the oxygen status of the filter systems began 15

weeks after planting, when the plants were about 0.8-1.3 m high and had till-
ered to around 20 shoots per wetland.
Oxygen concentration was measured with a Clark-type sensor designed at
the Heyrovsk:~ Institute of the Czechoslovak Academy of Science and pro-
duced by Chemoprojekt in Prague. The sensor consists of a Pt cathode 0.1 mm
in diameter and an Ag/AgCl anode; it produces a current of about 2 nA in water
saturated with air at about 20 ° C. The 02 consumption of this sensor is so low
that the difference between the signal in stirred and unstirred water does not
exceed 2 %. Therefore, the sensor can be used for in situ measurements without
stirring.
The calibration of the sensor tended to be unstable during the measurements
of some samples with low oxygen concentration. The instability can be ex-
plained by interference of H2S both with the Ag/AgC1 anode and Pt cathode.
Another calibrated sensor was used when instability occurred. The membrane
and the electrolyte of the poisoned sensor were replaced, the cathode was cleaned
and the sensor was recalibrated after its recovery. Frequent recalibration is
required when oxygen is measured in wastewater or other environments con-
taining H2S.

Diel fluctuations in oxygen concentration

Oxygen concentrations in situ in the filters were measured by inserting the

sensor into the filler tubes (Fig. 1 ) immediately before wastewater addition.
All the buckets were monitored giving a total of 48 readings at each observation
time.
308

Net oxygenation

Subsequently, some of the filters were flushed with at least three times their
volume of wastewater, which had been bubbled with nitrogen gas to reduce
oxygen concentrations to 3-4 mg 1-1. The net increase in oxygen concentration
was monitored over the following 68 h, with no further addition of wastewater.
This increase reflects the effects of rhizosphere oxygenation, together with any
re-aeration from the atmosphere, minus consumption of oxygen by the sub-
stratum. Observations were made on three replicate buckets from six treat-
ments. These were planted and plant-free systems amended with the following
combinations of lactic acid (L) or Typha leaves (T) : L - T - ; L + T - ; L +
T+.

Respiration

Respiration was measured using wastewater from the same series of filter
systems. Wastewaters were quickly aerated by rapid stirring and then confined
in full gas-free bottles which were incubated in the dark at a constant temper-
ature of 24 ° C. Oxygen concentration was monitored during the following 96 h.
Preliminary experiments showed that effluents from some of the systems
respired very slowly. These were inoculated with a further quantity of lactic
acid, equivalent to a carbon concentration of 25 mg 1-1, or with wastewater (20
ml ) from a system which gave a high rate of respiration.

RESULTS

Diel fluctuations in oxygen concentration

Oxygen concentrations measured in situ in the filters during flow-through

operation are shown in Table I and Fig. 2. Diel changes were generally rather
small, suggesting that an equilibrium concentration was reached within 9 h
after addition of the aerated influent. The presence of plants significantly in-
creased the oxygen concentrations in all the treatments except in filters with
no carbon amendment. Here, oxygen concentrations were high whether planted
or not, due to the low biological oxygen demand in the system. All planted
wetlands maintained oxygen concentrations above 8 mg 1-1, and were not sig-
nificantly different from each other. Except for the unsupplemented control,
all the unplanted systems gave low oxygen concentrations, ranging from 1.0 to
3.2 mg 1-1, reflecting the increase in respiration caused by supply of organic
carbon. Lactic acid was slightly more effective than ground leaves in reducing
oxygen concentration, and about 50 times more effective per unit of added
available carbon (Fig. 2 ).
Evidently, in planted treatments, rhizosphere oxygenation was sufficient to
compensate for the respiratory demands which were enhanced by carbon
supplementation.
 309

TABLEI

Oxygen concentrations in the artificial filters, measured in the morning at 07.30 h (temperature
18-22°C) and in the afternoon at 16.40 h (temperature 2 9 - 3 5 ° C ) , with each reading being the
average from six wetlands

Treatment' Morning 2 A f t e r n o o n2

Plants L T % saturation mg 1-' % saturation mg 1-1

a - - - 80.1 7.38 88.9 6.60

b - + - 20.2 1.80 22.8 1.70
c - - + 35.6 3.20 13.9 1.00
d - + + 14.0 1.14 12.7 1.00
e + - - 91.0 8.13 79.5 5.90
f Jr + -- 93.5 8.57 85.8 6.90
g + - + 94.0 8.50 82.7 6.10
h + + + 89.9 8.12 82.8 6.10

LSD (P<0.05) 9.7 1.04 11.5 0.34

'Filterswere with ( + ) and without ( - ) supplementation with organic carbon as lacticacid (L)
or ground leaves of 7".domingensis (T).
2Aerated wastewaters were introduced immediately after readings were made, so that figures rep-
resent depletion of oxygen during 14 h for morning readings and during 9 h for afternoon readings,
together with mixing of residual wastewater.

I0- f g

e
8 h
a

5 -

I+ -
c
c,J
/ G

x
0
m

0 I °
L - 4-
+
T - -- ÷
+
Fig. 2. Oxygen concentration in wetlands supplemented with lactic acid (L) or ground leaves of
T. domingesis (T), measured in situ at 07.30 h, 14 h after the previous addition of aerated was-
tewater. ~ =unplanted; EZ] =planted. Vertical bar gives L S D (P<0.01). Symbols (a)to (h)
correspond to treatments described in Table I.
 310

- Plants + Plants

-L-T ~ ' S

* i L I i i 0
C
+L-T b

i i i i i
/
.1_.3" h/J"
~" td i ~ - ' f

80
Time ( h i

Fig. 3. Changein oxygenconcentrationwith time measuredin situ in the filters,after introduction

of low-oxygenwastewater.Verticalbar givesrangeof duplicate measurements.Dotted line indi-
cates a singlemeasurement.Symbols (a) to (h) correspondto treatmentsdescribedin Table I.

Net oxygenation

Figure 3 shows the net oxygen production in selected wetland filters, follow-
ing flushing with low-oxygen wastewater. In planted wetlands with no carbon
supplements, or with only lactic acid addition, the oxygen concentration in-
creased rapidly, stabilising after about 20 h. In the presence of both carbon
sources there was a decline in oxygen concentration for 24 h, followed by an
increase. The increase in the net oxygen concentration reflects the release of
oxygen from the plant roots, together with any atmospheric re-aeration, minus
the consumption of oxygen through microbial respiration. In unplanted wet-
land filters with carbon addition there was little change over 68 h. The extent
of oxygenation, reflected in the net oxygen flux into deoxygenated wastewater
(Fig. 3 ), paralleled the trends of the oxygen concentration measured in routine
operation during a 6-day retention time (Fig. 2), confirming the important
role of the plants in maintaining an oxygenated substratum.

Respiration, carbon supply and microbial activity

The decline in oxygen concentration caused by respiration of wastewater

microorganisms, which was determined in bottle incubation experiments, is
shown in Fig. 4.
The addition of carbon substrate increased the respiration rate, ground leaves
(T) being more effective than lactic acid (L). Also, with no carbon amend-
ment, respiration rates were faster in wastewater from planted filters, than
from unplanted filters.
Figure 5 shows the effects on respiration of either added microbial inoculum
(I; of wastewater from actively respiring treatments) or of additional lactic
acid supplements (L).
 311

- plants +plantl +/-L +1-I

0 25 50 75 '100 0 25 50 75 25 5O 75 100

-L-T

J i
C ' ' ~ [0 --L -'11"
I
+L-T

-plants
o

-L+T

-I.plants
0 i ~ 10--L - T
0 25 50 75
T i m e (h)

+L+T 5 5

•I-plant s
0 tt.-'1"
255075 25 50 75 ~00 25 5O % 100
Time (h) Time (h)

Fig. 4. Change in oxygen concentration with time during incubation of aerated effluents from
wetlands in bottle experiments. Vertical bar gives range of duplicate measurements. Symbols (a)
to (h) correspond to treatments described in Table I.

Fig. 5. Effect of either added inoculum (I) or additional lactic acid supplement (L) on respiration
of effluents in bottle incubation experiments. • • -- without supplement; C) © = with
supplement. Symbols correspond to treatments described in Table I.

Respiration, or biological oxygen demand, must be satisfied by atmospheric

and rhizosphere oxygenation if the substratum is to remain oxygenated. Car-
bon supply, through its direct effect on respiration, is therefore expected to be
an important influence on the performance of artificial filters for retaining
metals or nutrients.
Results of the respiration experiments suggest that in unplanted, una-
mended wetlands the respiration rate was limited by carbon supply rather than
by a lack of adapted microbial populations (Fig. 5a). The reverse applied where
unplanted wetlands had been amended with lactic acid initially (Fig. 5b). Here,
the additional lactic acid increased respiration after a lag phase, perhaps when
microbial populations increased sufficiently to use the additional carbon.
In planted filters with lactic acid addition, the microbial inoculum was prob-
ably not limiting as it did not increase the respiration rate ( Fig. 5f).
Where there were no carbon supplements, effluents from planted filters res-
pired more quickly that effluents from unplanted filters, suggesting that bio-
logical activity was being promoted by carbon supplied in root exudates (Fig.
4). But, as demonstrated in Fig. 5e, the addition of extra lactic acid substan-
tially increased the consumption of oxygen even further.
312

T A B L E II

S u m m a r y of rates of net oxygen production measured in situ, consumption (respiration) meas-

ured in bottle incubation experiments and gross oxygenation calculated from the sum of net pro-
duction and c o n s u m p t i o n

Treatment 1 Net 02 02 Gross oxygenation

production consumption
Plants L T (pg h -1 (#g h -1 pg h 1 pg 1-1 h-1
filter 1) filter-l) filter-1

a - - m w
720 7 727 265
b - + - -70 81 11 4
d - + + -98 326 228 119
e + - - 550 195 745 325
f + + - 510 248 758 337
h + + + -40 480 440 203

1For details of t r e a t m e n t s , see Table I.

These results are encouraging in confirming the validity of the experimental

technique in that microbial colonisation was not inhibited in synthetic was-
tewaters containing metals at these concentrations.

Rhizosphere oxygenation: quantification

The oxygenation of the filters can be estimated from the rate of net oxygen-
ation (arbitrarily estimated after 20 h; Fig. 3) added to the oxygen consumed
to satisfy biological oxygen demand (Fig. 4). These figures (Table II) dem-
onstrate that, in planted treatments, gross oxygen production from rhizo-
sphere oxygenation and atmospheric exchange always exceeds oxygenation in
unplanted controls from atmospheric exchange alone. The difference, due to
rhizosphere oxygenation, was 747 pg h -1 in treatments amended with lactic
acid, and 212 ttg h -~ in treatments amended with both ground leaves and lactic
acid or 333 and 84 pg 1-1 h -1, respectively. These values are probably underes-
timates because the system was static during re-aeration, allowing the greater
development of non-stirred diffusion zones which could limit overall oxygen-
ation compared with a flowing system. Also, respiration was determined using
effluent from the wetlands, so that the biological demand of the microbial films
left undisturbed in the substratum was not included in the estimation. In un-
supplemented filters,the gross oxygenation in the planted system (745 ttg h -1)
was only slightly higher than in a system without plants ( 727 ttg h -1) so that
rhizosphere oxygenation, by difference, was only about 18 pg h - 1. This reflects
the low demand of these systems for oxygen. In the unsupplemented system,
the plant supplied only its own oxygen requirement and was not required to
 313

satisfy a demand for oxygen created by bacterial degradation of lactic acid or

ground Typha leaves.

Metal retention: effect of pH, redox and rhizosphere oxygenation

Table III shows the partitioning of applied metal loads during routine flow-
through operation between 4 and 18 weeks after planting. The quantity re-
tained by the gravel substratum was calculated by difference: the sum of the
metal loads leaving in the outflow, harvested in living plants and scavenged by
ground Typha leaves was compared with the metal loads applied.
Table IV shows the proportion of influent metal retained in the filters (metal
retention), together with redox status and pH. Values for metal retention were
calculated from the difference in daily metal loads flowing into and out of each
filter. For example, a metal retention of 80% means that 80% of the influent
metal load is distributed between the plants and/or substratum within the
filter, and the remaining 20% leaves the filter in the effluent.
TABLE III

Influent and effluent metal loads, and metal uptake into T. dorningensis plants and ground leaf
amendments at harvesting. (Quantities of metal retained in the substratum were calculated by
difference)

Metal Treatment 1 Metal partitioning ( m g ) 2

Plants L T Influent Plant Ground leaves Substratum Effluent

Cu C -- - + 21.5 - 2.98±1.27 17.3 1.23±0.14

d - + + 20.6 - 2.04±0.20 17.4 1.19+0.14
e + 21.5 10.16_+0.39 - 9.9 1.48_+0.08
f + + - 20.6 9.55_+0.73 - 9.5 1.50 ± 0.08
g + - + 21.5 8.31 ± 0.52 4.02 _+0.54 8.5 0.66 + 0.05
h + + + 20.6 7.67±0.52 1.99±0.18 10.0 0.95_+0.11
Zn C -- - + 100.5 - 19.15_+0.76 78.3 3.11 ±0.41
d - + + 96.2 - 11.48_+1.23 81.7 3.04_+0.66
e + 100.5 38.29 ± 2.31 - 61.5 0.76 _+0.20
f + + - 96.2 31.90_+2.09 - 63.6 0.71 ± 0 . 1 2
g + - + 100.5 28.19 ± 1.81 17.26 ± 1.49 53.1 1.96 _+0.39
h + + + 96.2 25.72 _+ 1.49 11.00 ± 1.00 56.2 3.31 ± 0.61
Cd C -- - + 19.9 2.96 _+0.25 16.4 0.52 _+0.06
d - + + 18.8 2.08 ± 0.17 16.2 0.52 _+0.09
e + 19.9 4.73 ± 0.30 15.0 0.16 _+0.03
f + + - 18.8 4.54__0.27 14.1 0.13±0.02
g + -- + 19.9 4.09 -+ 0.28 3.00 ± 0.33 12.5 0.29 _+0.05
h + + + 18.8 3.87±0.26 1.94±0.19 12.6 0.45-+0.11

~For details of treatments see Table I.

2Figures are mean ± s.e.
314

Over all t r e a t m e n t s , t h e o r d e r o f m e t a l r e t e n t i o n in w e t l a n d s was directly

r e l a t e d to p H , a n d o n l y p o o r l y r e l a t e d to o x y g e n c o n c e n t r a t i o n a n d r e d o x po-
tential. In u n p l a n t e d wetlands, t h e effects of c a r b o n a m e n d m e n t s t r o n g l y in-
f l u e n c e d p e r f o r m a n c e efficiency, m a i n l y t h r o u g h t h e effects o n p H a n d r e d o x
p o t e n t i a l , r a t h e r t h a n t h r o u g h c h a n g i n g o x y g e n status.
T h e p r e s e n c e of p l a n t s i m p r o v e d m e t a l r e t e n t i o n in all w e t l a n d s , b u t p a r t i c -
u l a r l y in - L - T a n d + L - T t r e a t m e n t s ( T a b l e IV, t r e a t m e n t s e a n d f) w h e r e
p H was significantly increased. Analysis o f t h e p l a n t s ( T a b l e III) s h o w e d t h a t
o n l y a b o u t 2 0 - 4 7 % o f t h e m e t a l load was r e t a i n e d in t h e p l a n t m a t e r i a l , while
n e a r l y all t h e m e t a l inflow ( 9 7 . 2 - 9 9 . 9 % ) was s c a v e n g e d b y t h e p l a n t s a n d
s u b s t r a t u m t o g e t h e r . Also, w h e n b o t h p l a n t s a n d g r o u n d leaves were p r e s e n t
t h e y o n l y a c c o u n t e d for up to 58% of t h e m e t a l s r e m o v e d ( T a b l e s III a n d IV,
t r e a t m e n t s g a n d h ) , a l t h o u g h m e t a l r e m o v a l f r o m i n f l u e n t s in the s y s t e m s
overall was close to 100%. T h e r e f o r e , in p l a n t e d w e t l a n d s , t h e p e r f o r m a n c e
was c o n t r o l l e d b o t h b y u p t a k e i n t o t h e p l a n t s t h e m s e l v e s a n d also b y indirect
effects of t h e p l a n t s o n t h e p h y s i c o c h e m i c a l c o n d i t i o n o f t h e s u b s t r a t u m .
In u n p l a n t e d filter s y s t e m s t h e r e was a s u b s t a n t i a l i n c r e a s e in m e t a l r e t e n -
t i o n in the p r e s e n c e o f c a r b o n s u p p l e m e n t s ( T a b l e IV, t r e a t m e n t s b to d com-
p a r e d w i t h a ) . D i r e c t u p t a k e into t h e g r o u n d leaves o f T. domingensis was less
t h a n a b o u t 17% o f t h e m e t a l load ( T a b l e I I I ) , suggesting t h a t filter p e r f o r m -
ance was again i n f l u e n c e d s u b s t a n t i a l l y b y i n d i r e c t effects o f the a m e n d m e n t s
on t h e p h y s i c o c h e m i c a l c h a r a c t e r of t h e s u b s t r a t u m .

TABLE IV

Redox status, pH and retention of metals in artificial filters with ( + ) and without ( - ) supple-
mentation with organic carbon as lactic acid (L) or ground leaves of T. domingensis (T)

Treatment' Redox2 pH Metal retention'~

(mV) ( % of influent)

Plants L T Cu Zn Cd
a - - - 548 4.3 78.8 49.0 62.3
b - + - 203 6.0 83.5 93.6 96.2
c - - + 118 7.3 94.9 96.4 98.0
d - + + 86 7.8 91.4 96.0 98.0
e + - - 539 7.2 97.2 99.9 99.9
f + + - 558 7.1 97.8 99.9 99.9
g + - + 268 7.6 99.1 99.8 99.4
h + + + 216 7.5 99.2 99.3 99.9

LSD 78 0.4 4.0 4.8 31

(P<0.01)
'For details of treatments see Table I.
2Mean of five redox probes in one randomly selected wetland filter.
3Means of six wetlands each measured daily.
 315

The gravel used in these experiments was rich in iron and coarse in texture,
with a low native organic carbon content. In this neutral, oxygenated substra-
tum the mechanisms of metal removal are probably dominated by precipita-
tion as metal oxides, adsorption to colloidal hydrous oxides and chelation with
organic matter ( Patrick and Gambrell, 1976). Under these conditions the plants
were able to increase pH to neutral and counteract the respiratory demand
enhanced by organic carbon supplementation. In summary, the presence of the
plants was critical for the satisfactory performance of these artificial wetland
filter systems.

ACKNOWLEDGEMENTS

This research was funded by the Australian Mineral Industries Research

Association Ltd. For advice and helpful discussion we are grateful to several of
the sponsors, and also Dr. D.B, Nairn, Dr. D.S. Mitchell, P.R. Breen, D.L.
Short and A. Chick. Advice on statistical analysis and data processing were
given by Dr. Bob Forrester and D. Erskine. Analytical advice and help was
given by M.L. Higgins and W. Korth. R. Dalgleish gave superb technical
support.

APPENDIX I

Preparation of nutrient solution and metal-enriched synthetic effluent

Chemical compounds Compound in Dilution Final ionic

stock solution factor concentration
(g1-1 ) (mg 1-')
KH2P04 42.4 } (P) 9.6
K2S04 65.2 1000 (K) 41.4
NaC1 15.2 (Na) 6
(C1) 9
B NH4NO,~ 287.1 5000 (N) 20.1
Ca(NO3)2.H20 648 1250 (N) 79.8
(Ca) 114
C MgSO4.7H20 196 1600 (Mg) 12
(s) 28
D H3B03 2.86 } (B) 0.094
MnCI2.4H20 1.81 (Mn) 0.094
ZnSO4.7H20 0.22 5333 (Zn) 0.0094
CuSO4.5H~O 0.08 (Cu) 0.0032
(NH4) ~MOTO24.4H20 0.131 (Mo) 0.0019

E Sequestrene 138 Fe' 13.65 533 (Fe)

1Fe supplied as sodium ethylenediamine di- (o-hydroxy phenyl acetate).
316

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