Pergamon Chemosphere, Vol. 38, No. 13, pp.

3155-3168, 1999
© 1999 Elsevier Science Ltd. All rights reserved
0045-6535/99/$ - see front matter

PII: S0045-6535(98)00522-0
EFFECTS OF CADMIUM ON THE PERFORMANCE AND MICROBIOLOGY OF
LABORATORY-SCALE LAGOONS TREATING DOMESTIC SEWAGE

J.L. B o n n e t *a , J. Bohatier a - b a n d D. P~pin c

aLaboratoire Biologic Compar4e des Protistes, UPRESA CNRS 6023, Universit~ Blaise Pascal, 63177 AUBIERE Cedex, FRANCE

bLaboratoire Biologic Cellulaire, UFR de Pbarrnaeie, BP 38, 63001 CLERMONT-FERRAND Cedex, FRANCE

CLaboratoire Hydrologic Hygiene et Environnement, UFR de Pharmaeie, BP 38, 63001 CLERMONT-FERRAND Cedex, FRANCE

(Received in Germany 29 April 19987; accepted 23 September 1998)

ABSTRACT

Two experiments were performed to assess the impact of cadmium on the sewage lagoon wastewater
treatment process. For each one, three laboratory-scale pilot plants with one tank receiving the same raw
effluent were used; one plant served as control and the other two were contaminated once only with cadmium.
In the first study, the effects of a shock load of two concentrations of cadmium chloride (60 and 300 pg/1) on
the plant performance, microbial populations (protists and bacteria) and enzyme activities were determined.
Initially, most of the performance parameters were affected concentration-dependently. A reduction in the
protist population density and some influence on the total bacterial population were observed, and the potential
enzymatic activities were also modified. A second experiment with a lower cadmium concentration (30/~g/l),
supplied as chloride or sulphate, still perturbed most of the parameters studied, and the effects of the two
cadmium salts were identical. © 1999 Elsevier Science Ltd. All rights reserved

Key words: cadmium, pollution, sewage lagoon process, wastewater treatment, laboratory-scale pilot plants.

INTRODUCTION

Cadmium, an industrial pollutant, ranks among the most toxic heavy metals [1-5]. It can form stable
combinations with SH groups and has been responsible for Itai-Itai disease, gastro-intestinal and pulmonary
disorders, and cell and tissue alterations [6].
Numerous environmental factors influence cadmium toxicity, including nature of medium, nature of
sediment, organic matter content, pH, dissolved oxygen, and speciation of others metals [3,7]. Research has
shown that cadmium is toxic to aquatic organisms: algae, cyanobacteria and protozoa are the most sensitive,
whereas bacteria, fungi and some invertebrates are the most resistant. For bacteria, the concentrations
necessary to induce a toxic effect range widely, and several resistance mechanisms have been chamcterised
[2,8,9]. For phytoplankton, effects have been determined on growth, morphology, photosynthesis, and
3155
3156

nutrient use [3,4]. Negative effects of cadmium (physiology, ultrastructure) have been shown in protozoa [10-
14]. Some studies have also shown a toxic action on crustaceans, cladocerans particularly [3,15,16].
The widespread industrial use of cadmium means it can be found in natural aquatic environments and
wastewater treatment systems, where it can cause toxic effects and reduce the efficiency of biological
treatments. As a recognised source of pollution, its ecotoxicological impact needs to be known precisely.
However, most of the research carried out on the toxic effects of cadmium on aquatic ecosystems have so far
studied single aspects, such as the influence on micro-organisms, water quality or metal behaviour [ 1,17-19].
To our knowledge, no work has addressed the overall effects on a wastewater treatment process, and
specifically on a sewage lagoon process. The aim of this work was therefore to study the influence of
cadmium on plant performance, microbial populations (protists and bacteria) and enzyme activities of the
micro-organisms by means of laboratory-scale pilot plants mimicking a full-scale lagoon process. The natural
lagooning process was chosen as an aquatic model because it is the treatment system that possesses the highest
degree of homology with the natural aquatic ecosystems. We have also studied the behaviour of cadmium by
determining the time course of its concentration in the water column and outflow effluent.
In previous work [20], we demonstrated that our laboratory pilot plants, in particular the one-tank
system, accurately simulate the operation of a natural sewage lagoon plant after an initial stabilization period of
about one month. Statistical analysis has revealed a similarity of operation and an equivalent depollution
capacity for the three-pond reference plant and laboratory plants using one or two tanks. Overall, the results
have shown that the first pond (reference plant) or tank (laboratory systems) take the main share in the
depollution process for all the parameters studied.

MATERIALS AND METHODS

a) Cadmium assay
Cadmium was used as chloride (Sigma Chemicals C-2544) and sulfate (Sigma Chemicals C-2919)
dissolved in deionised distilled water. Final concentrations were 30, 60 and 300/,~g/1.
Cadmium concentrations were determined by atomic absorption spectrophotometry using a graphite
furnace with Zeeman effect (Zeeman 500 Perkin-Elmer) for contents below 50 ,ug/1 and by emission
spectrometry (Inductive Coupled Plasma analysis) (ICP 6500 Perkin-Elmer) for contents above 50 ug/1.

b) Laboratory pilot plants

Laboratory-scale pilot plants mimicking a full-scale lagoon process have been described previously
[20]. For each experiment, three systems with only one tank were run simultaneously. In the first experiment,
after the initial stabilization period (45 days), pilot "Pil 1" without cadmium served as control, pilot "Pil 2"
received once 60/~g/l CdC12 at the tank head, and pilot "Pil 3" received once 300/~g/l CdC12 at the tank head.
In the second experiment, after the initial stabilization period (40 days), pilot "Pil 4" without cadmium served
as control, pilot "Pil 5" received once 30 ,ug/l CdC12 at the tank head, and pilot "Pil 6" received once 30 ,ug/l

C d S O 4 at the tank head.

e) Artificial substrates
In this study, polyurethane foam (PF) blocks were used as artificial substrates to collect the different
micro-organisms present in the medium. Small cubes (2 x 2 x 2 cm) were cut from PF, a three-dimensional
 3157

interstitial material. Several blocks, hanging on nylon yarn from a rod, were immersed at the same time in the
water column in the centre of the tank of each pilot. The PF blocks were installed in the pilots one week before
adding the cadmium.
The blocks were withdrawn three at a lime from each pilot at regular intervals. The first three were
taken out after 7 days and the last three at about 1 month. About 7 ml was collected by squeezing. A 1-ml
sample was taken immediately for ciliate determination. Each block was then rinsed with 30 ml of distilled
water. They were squeezed a second time to collect all the block content. From this volume, a 5-ml sample
was used for flagellate determination and a 30-ml one for chlorophyll pigment assay.

d) Physico-chemical and biological parameters measured

Chemical oxygen demand (COD), biochemical oxygen demand (BODs), concentrations of N-NH 4, N-

NO 3, N-NO 2, and P-PO 4, pH, conductivity, dissolved oxygen, volatile suspended solids (VSS) and
suspended solids (SS) were measured according to the techniques described previously [20].

e) Determination of the different micro-organisms

Total non-filamentous bacteria were determined by epifluorescence microscopy with the acridine
orange technique and the micro-organisms were counted by means of an image analysis system [20,21].
The biomass represented by the phytoplankton was estimated by assay of chlorophyll pigments. The
samples were collected from the liquid associated with the PF blocks as described under "artificial substrates".
The assay was carried out by spectrophotometry according to the procedure employed previously [20].
Ciliate protozoa were recovered from the liquid collected from the PF blocks. Living micro-organisms
were identified and(or) counted under binocular magnification with a Dolfuss cell.
Heterotrophic and autotrophic flagellate protozoa were also collected from the liquid associated with the
PF blocks. They were counted by epifluorescence microscopy with the primulin technique [22]. Primulin
allows us, on the same preparation, simply by changing the filter, to distinguish between plastid and non-
plastid flagellates. The autotrophic flagellates were characterised by a red-orange autofluorescence of
chlorophyll a [22].

f) Potential enzymatic activities

Nitrification activity was measured by the method of TOMLINSONet al. [23] modified by NAHIMANA[24]
and previously described [25].
Denitrification activity was measured by the method of NAKAJIMAet aL [26] and HOCHMANet al. [27]
modified by NAHIMANA[24] and previously described [25].
Esterasic activity was measured by the method described for activated sludges by FONTVlEILLEet al.
[28] and modified by BONNET ]21] for sewage stabilization ponds. The protocol consists in the hydrolysis of
fluorescein diacetate and a spectrophotometric quantification of the free fluorescein.

g) Statistieal analysis
Variance analysis (F test) was performed using Statview software on most of the parameters studied.
This statistical analysis was carried out to check for any significant difference between the state of the three
pilots for a given parameter at the end of the stabilization period. This test consisted in comparing calculated F
with F values from the Fisher-Snedecor table for a significance threshold. If calculated F was greater than the
3158

theoretical value o f the table at the threshold chosen (5% most often), the difference was significant.

RESULTS

A- I n f l u e n c e o f 6 0 a n d 300 /ag/I of CdCI 2

The results o f variance analysis (tablel) show that some parameters were similar in the three pilots at
the end o f the stabilization period (day 45), whereas others were significantly different between two or three
pilots.

Parameters calculated F Significant difference Pilots concerned

N-NO3 0.147 no
N-NO 2 0.429 no
N-NH4 0.080 no
P-PO4 0.440 no
COD 1.830 no
BOD5 0.900 no
Temperature 0.040 no
pH 32.158 yes 1/2 1/3
Conductivity 1.997 no
Dissolved oxygen 30.327 yes 1/2 l/3 2/3
Suspended solids 4.418 yes l/2
Total bacteria 58.853 yes 1/2 l/3 2/3
Esterasic activity, 58.484 yes 1/3 2/3
Table l: variance analysis (F test) for the pilots Pil 1, Pil 2 and Pil 3 on the stabilization period days 28 - 45
(parameters measured in the water column).

a) Effects on p l a n t p e r f o r m a n c e
The addition of cadmium to pilots Pil 2 and Pil 3 caused increases in COD, B O D 5, nitric nitrogen, and
nitrous nitrogen values (fig. la to ld). These concentration-dependent increments were strong and prompt
during the first days and then more gradual after 14 days. For some parameters, we observed a gradual return

to initial values. For ammoniacal nitrogen and P-PO 4 (fig. le and lf), the effects seemed slight. For the
control, the values remained stable throughout the experimental period. Overall, we noted a negative effect on
the treatment efficiencies in the treated systems compared with the control.
An effect was also noticeable for pH values and dissolved oxygen concentrations. The pH values
increased more in the treated pilots than in the control. The dissolved oxygen concentration decreased strongly
and sharply after the Cd addition, and a gradual return to initial concentrations was observed after l 0 days
(fig. 2b). T h e control values displayed only slight variations throughout the experimental period.

b) B e h a v i o u r o f c a d m i u m
Assay o f cadmium (fig. 2a) s h o w e d that the raw effluent continuously added to the pilots was
essentially free o f cadmium. At the water column and the outflow levels o f the treated pilots, we noted a
prompt decrease in the free cadmium concentration against time, followed by a more gradual reduction. After
14 days, the concentrations were low at all the experiment points. For the control pilot, only traces of
cadmium were found throughout the experimental period.
)~ N-NH 4 (maql) N-NO (r~8;l) C O D (zn~4|)

~: !!!

Z p-PO. (mgn) N-NO z (m~t) B O D s (rosa)

¢,~ N N
N

~ N N

N ' N
3160

3oo

25o

2oo

150

,oo

5 o ~ a
o ..... ~ , , Q
0 7 14 21 28 35 (days) 42

~ Raw e f f l u e n t & Pil 1 w a t e r ~ P i l 1 outflow : Pil 2 w a t e r

-- -0- -Pil 2 outflow "-- Pil 3 w a t e r ---I-I--- Pil 3 o u t f l o w

80

Controlpilot

i--o- ~ t a0o~l ca I

o~,O 6o

co~
2

b
20 ~ I
10 20 30 40 50 60 70 $0 90 0 10 20 30 40 50 60 70 g0 90
(days) (days)

20~ I1 Controlpilot I 30

,s . . . . / m .,°t 3°°.g., CdI

IO
s 1 5

d e
0 o
0 l 3 7 10 14 21 42 0 l 3 7 lO 14 21 42
(days) (days)

Fig. 2: Time course of cadmium (a), dissolved oxygen (b) and potential enzymatic activities (c to e)
during the experiment period. On graphs a, d and e, stabilization period is not shown (day O is the day
of cadmium addition to the pilots).
 3161

c) Effects on t h e m i c r o b i a l p o p u l a t i o n s
The total bacterial population fluctuated non-significantly after the addition o f cadmium. In all cases
(control and treated pilots), a marked fall was noted between the raw effluent and the outflow from the pilots.
Autotrophic and heterotrophic flagellates were counted from the liquid associated with the P F units.
Time course s h o w e d a concentration-dependent decrease in the population density compared with the control
in which the colonization seemed to be maximal after 21 days.
Identification o f the dominant ciliate species that colonized the blocks indicated a less significant effect
between control and treated pilots.

d) Effects on the potential m e t a b o l i c activities of the m i c r o - o r g a n i s m s

For this first experiment, three activities were measured (fig. 2c to 2e): nitrification, denitrification, and
esterasic activities. The nitrification activity decreased after 24 h in Pil 3, and after 72 h in Pil 2. The activities
then remained level as those o f the control pilot until the end o f the experiment. An increase in the
denitrification activity was noted from day 10 in the contaminated pilots. The esterasic activity was enhanced
very soon after the addition o f cadmium. This enhancement persisted thereafter at a lower level.

B- I n f l u e n c e o f 30 pg/I o f CdCI 2 or C d S O 4
The results o f variance analysis (table 2 and 3) indicated similar states for the three pilots at the end o f
the stabilization period for all the parameters studied, except for esterasic activity.

Parameters calculated F Significant difference Pilots concerned

N-NO3 1.445 no
N-NO 2 0.130 no
N-NH4 0.007 no
P-PO4 0.045 no
COD 3.368 no
BOD5 2.333 no
Suspended solids 1.301 no
Volatile suspended solids 1.909 no
pH 0.388 no
Dissolved oxygen 0.552 no
Conductivity 1.818 no
Total bacteria 2.439 no
Esterasic activity, 40.121 yes 4/5 4/6 5/6
Table 2: variance analysis (F test) for pilots Pil 4, Pil 5 and Pil 6 on the stabilization period days 27 - 40
(parameters measured in the water column).

Parameters calculated F Significant difference Pilots concerned

Autotrophic flagellates 0.022 no
Heterotrophic flagellates 0.075 no
Chlorophyll a 0.002 no
Ch/orophyll b 0.051 no
Chlorophyll e 0.176 no
active Chlomphyll a 0.258 no
Pheo-pigments 0.009 no
Carotenoids 0.270 no
Table 3: variance analysis (F test) for pilots Pil 4, Pil 5 and Pil 6 on the stabilization period days 27 - 33
(parameters measured from the liquid associated with the polyurethane foam blocks).
3162

a) Effects on plant performance

The addition of cadmium to pilots Pil 5 and Pil 6 caused, as in the first experiment, an
enhancement of the values of COD, BODs, nitric and nitrous nitrogen. For these parameters, no
significant differences were detected between the two salts of cadmium used at 30 ~g/1. As expected,
the gradual reduction of the treatment efficiencies in the treated pilots was weaker than in the presence
of 60 or 300/~g/l of Cd. Dissolved oxygen and pH displayed similar profiles to those observed in the
first experiment, but the magnitude of the variations appeared to be lower. Concerning suspended
solids and volatile suspended solids, the effects generated by the two cadmium salts seemed to be quite
different. An increase was noted with both the chloride and the sulphate, but sulphate caused a greater
enhancement of the values.

b) Behaviour of cadmium
The same type of results were obtained as in the first experiment. A decrease in the free cadmium
concentration against time was observed. Those decreases that were prompt in the first part of the
experiment were similar with the two cadmium salts used.

e) Effects on the microbial populations

Although the total bacterial population did not vary significantly after the addition of cadmium,
this was not the case for the various populations of protists (fig. 3 and 4). The cadmium caused a
decrease in the ciliate, and autotrophic and heterotrophic flagellate population densities. The effects
were practically the same with the two salts employed. For the flagellate populations, the effect was
weaker than in the first experiment.
In this second experiment, the liquid from the PF blocks was also used for the determination of
the different chlorophyll pigment concentrations. The first experiment showed that samples taken
directly from the water column did not permit the effects of cadmium on the phytoplanktonic
populations to be accurately assessed. A continuous increase occurred for each pigment (chlorophyll a,
chlorophyll b, chlorophyll c, active chlorophyll a, carotenolds, pheo-pigments) in the control pilot, In
the case of the treated pilots, the concentrations were lower to those determined for the control pilot
according to the type of pigment considered. For the same pigment, the two salts induced identical
effects. We only showed the evolution of one pigment (fig. 4c).

i,o, 1
t. R~weffio~ot
t
e ~tcdc~ I ~oo
Pilot CdS04

a b
10 , i , i , i , i , i , i , i__

10 20 30 40 50 60 (days) 70 7 14 21 (d~ys) 28

Fig, 3: Time course of total bacterial (a) and ciliate (b) populations during the experiment period. On
graph b, stabilization period is not shown (day 0 is the day of Cd addition to the pilots).
 3163

14
g ---13-- Controlpilot ~ Controlpilot I~ ]
..... ......... /

7 14
......... :f
21 (daTs) 28 (days)

"~ ~:: r;dl3pi;t

20O00 . . . . . .

3,0 . . . . . .

25O .--0

50O00 / c 2'0 d

3 10 17 21 (days) 28 , , ~ * ~ , ~ , , . ~ , ~ , =
0 lO 20 30 40 50 60 (days) 70

Fig. 4: Time course of flagellate populations (a and b), chlorophyll a concentration (c) and esterasic
activity (d) during the experiment period. On graphs a, b and c, stabilization period is not shown (day 0
is the day of cadmium addition to the pilots).
d) Effects on the potential metabolic activities of the micro-organisms
For this second experiment, only esterasic activity was measured (fig. 4d). The time course was
identical for cadmium chloride and sulphate. We observed a sharp increase in this activity after the
addition of cadmium, followed by a stabilised state in the second part of the experiment.

DISCUSSION

Wastewater treatment systems have to deal with organic and(or) inorganic contaminants of widely
diverse origins, and so knowledge of the microbiological and physiological effects such substances
produce in these systems, and their impact on treatment efficiency, is of the utmost importance. We
used laboratory pilot plants that mimic wastewater treatment by the natural lagoon process to study the
impact of cadmium pollution on the whole of the treatment process. Only an approach based on a
broad range of methods (analytical, microbiological, enzymological .... ) can afford a full understanding
of the impact on aquatic environments. Yet most of the studies conducted to date concerning the
impact of metal ions on treatment systems have been limited to comparisons of input-output balances
or the monitoring of some particular aspect [ 17,18,29,30].
3164
The two successive experiments carried out with cadmium showed a reduction of decontamination
capacity relative to a control system free of cadmium ions. We found a concentration-dependent increase with
time in the values of the main parameters that quantify treatment efficiency in the pilot plants. However, the
results do not show wide variations in amplitude for the different Cd concentrations used (30, 60 and 300
,ug/l). The impact was weakest with 30/ag/l, and was the same for cadmium chloride and sulphate for most of
the parameters. These experiments confirm the toxicity of cadmium already observed in different environments
[I,2,4,5,7,11,19].
Concerning the fate of the cadmium, the concentrations fell rapidly in the water column in the Cd-
treated pilots during the first days, and then more gradually. After 14 days, practically no cadmium could be
found in the pilots. Also, a certain amount of cadmium left the pilot systems within the first 24 h. After
introduction into the pilots the cadmium is found in the different compartments of the system; in solution in the
water column, adsorbed or captured by micro-organisms, trapped by various suspended particles, bound to
sediment (either by direct binding from the dissolved fraction, or by particle settling). The bioavailability and
speciation of the cadmium in the medium are important parameters that determine its toxicity towards micro-
organisms [1,3,7]. In future experiments, it will therefore be interesting to determine precisely what the
concentrations of cadmium are in each fraction, to assess its toxicity more fully.
In each of the two experiment conducted, we observed an increase in suspended solids and volatile
suspended solids in the Cd-treated pilots. This was not unexpected, given the reduction of treatment efficiency
and the ensuing increase in organic matter present. A lack of correlation was observed between suspended
solids (or volatile suspended solids) and bacterial biomass. Although these values are widely considered to be
representative of the biomass, our results show that this is not the case; suspended solids and volatile
suspended solids are not specific to the biological compartment, since they account for both living and inert
matter (dead cells and particles with no biological activity). This means that in enzymological studies, the
expression of results in terms of volatile suspended solids undervalues enzyme activities.
Concerning the different micro-organisms present, it is difficult to evaluate and separate the effects
produced by the different populations from those that concern the whole of the ecosystem, because a
xenobiotic acts simultaneously at many levels. Reduced abundance and variety of species in aquatic media
exposed to pollution are the most frequently observed effects. Reduced diversity of microbial communities in
an ecosystem reflects general environmental damage.
Although the total bacterial count did not vary significantly after addition of cadmium in the two
experiments, this result may nevertheless be considered as the consequence of several concomitant effects.
When placed in contact with bacteria, a toxic substance can exert an inhibiting action, be biodegraded,
bioconverted or mineralised, or remain inert. Cadmium is known to be toxic to most bacteria, but the
concentration thresholds for toxic effects range widely, from ,ug/l to mg/l [1-3]. The constant total numbers of
bacteria found in our experiments may therefore be due either to a general lack of effect of the different
concentrations tested, or more probably to a selective effect on the bacterial flora with elimination of sensitive
strains and selection of resistant strains. Acquired cadmium resistance is well documented [2,8,9,31]; Gram-
negative bacteria are known to be generally more resistant to cadmium [3]. Our results show that overall
bacterial impact is not sufficient to account for a specific effect of a xenobiotic. For a more thorough
understanding of the disturbances generated, the fates of the different groups of bacteria present have to be
monitored. Immunological probes prepared against defined bacterial strains, already used in an activated
sludge system in a study of the impact of chromium and copper, are one possible approach [24]. Other
 3165

methods, such as numerical taxonomy [32], ribotyping [33] or nucleic probes [34], might also help to study
the evolution of a given group of bacteria in this ecosystem following a pollution challenge.
Microalgae, flagellates and ciliates were affected by the presence of cadmium in the pilots. This result
is consistent with the fact that protists are among the most cadmium-sensitive micro-organisms [2]. In the
lagoon type system, the diversity of the eukaryotic microbial populations is broader than in other wastewater
treatment systems, but the biomass present in the water column is lower. The study of the disturbances caused
by cadmium on these populations was possible only by the use of an artificial substrate (PF blocks) to collect
objectively the microfauna. As a true microcosm, this artificial substrate was successfully used previously for
stagnant and running water to study diverse microbial communities [35,36]. The use of PF blocks enabled us
to evaluate, qualitatively and quantitatively, the eukaryotic microbial communities in the control and Cd-treated
pilots. Although the microbial densities determined in the liquid phase from the PF blocks are very different
from the real values determined in the water column (certainly owing to the absence of predators in the
blocks), they will nevertheless reveal any influence of a toxic substance by comparison with a control. In this
way we showed a concentration-dependent reduction of the colonising capacities of the ciliates and flagellates
when the pilot was treated once with cadmium. These results agree with those of FERNANDF.Z-LEBORANSand
NOVlLLO [13,19,37], and MA~N! et al. [12,18,38], who report disturbances in communities of protozoa
exposed to cadmium.
Various effects of small concentrations of cadmium on growth, morphology, photosynthesis, and
nutrient absorption and assimilation in phytoplankton were reported [1-4,39]. In our first experiment, the
effect on phytoplankton, estimated by assay of chlorophyll pigments in water column samples, could not be
determined precisely; decreases in low concentration values have to be interpreted with great caution. In the
second experiment, we showed that levels of chlorophyll pigments, i.e., of total phytoplankton biomass fell in
the Cd-trcated pilots, by measurements in liquid collected from the PF blocks. All the autotrophic populations
were disturbed by the presence of cadmium, because the concentrations of all the types of chlorophyll pigment
decreased. However, it seems that the micro-organisms that are rich in chlorphyll b, chlorophyll c and
pheopigments are preferably affected. After addition of cadmium, dissolved oxygen levels fell and then
gradually reverted to initial values. This result agrees with those concerning the variations in autotrophic
populations and variations in the concentrations of chlorphyll pigments, the fall in dissolved oxygen level
results directly from the effects of cadmium on the phytoplanktonic organisms, which are mainly responsible
for the oxygenation of the water. The gradual return to the initial state, and the correlated increase in pH
observed at the end of the experiment, suggest that the phytoplanktonic micro-organisms return to
approximately normal physiological activity under these experimental conditions.
To our knowledge, no other study as complete as ours on the impact of cadmium on all the
communities present in an ecosystem of the natural lagoon type has ever been conducted. In recent years,
various studies including several trophic levels have been carried out, but on other media, to study the impact
of cadmium. A study of the regulation of phytoplankton and zooplankton biomass in rockpool food webs
under chronic cadmium pollution [40] showed that cadmium (at 20/~g/1) had a negative effect on all trophic
levels. Another study concerning the regeneration, recycling and trophic transfer of trace metals (Zn and Cd)
by microbial food-web organisms in the pelagic surface waters of Lake Erie [41] showed that picoplanktonic
organisms, which are ideally suited to the scavenging of trace metals, were grazed heavily by
microzooplankton. This grazing resulted in both a trophic transfer of these metals and their regeneration in the
dissolved phase. For cadmium, recycling was more important than trophic transfer. This study also showed
3166

that microzooplankton grazing tends to prolong the residence times of these metals in the pelagic surface
waters. A similar study should enable us to determine with greater precision the cadmium cycle in a natural
lagoon wastewater treatment plant, and the exact role of each community of micro-organisms in this cycle; this
can be done in parallel with the study of the fate of the cadmium mentioned above.
We also focused on certain potential enzyme activities of micro-organisms that can be used to assess
the physiological state of the cells after addition of a xenobiotic. Cadmium, which has no biological function
of its own, is known to impair many enzyme activities [2,3]. In our two experiments, a single introduction of
cadmium in the pilots caused first a sharp, and then a gradual increase in esterasic activity, an indicator of cell
metabolism [28,42,43]. This therefore points to an increased cellular metabolic activity. It is difficult to
interpret these results more precisely, because hydrolysis of the substrate used (fluorescein diacetate) can arise
from reactions catalysed by enzymes of different types (esterases, lipases, serine proteases), that may take
place inside or outside the cells [28] or exclusively inside the cell [42,43]. The values measured only give
general indications about the metabolic activity of the micro-organisms. The results observed are surprising,
however, because an inhibition of this activity was generally found in the presence of different metals (Cd,
Zn, Hg, Cu, Ni) in marine algae [42] and in a rotifer [44]. Inhibition of nitrification was observed in the two
Cd-treated pilots, but later with the lowest concentration of cadmium. This agrees with the work of NAHIMANA
[24], who also found an inhibition of this activity' in the presence of other metals (Cr and Cu) in an activated
sludge treatment system. Cadmium did not modify denitrification activities overall in the first part of our
experiment. However, a stimulation was observed from day 10. This results differ from those of NAHIMANA
[24]. The effects of cadmium on these two activities should logically lead to a strong decrease in nitric nitrogen
concentrations in the water column of the Cd-treated pilots. Assay results show the opposite. This
demonstrates the difficulty of linking potential enzyme activities that the micro-organism can express when
they meet an exogenous substratc, and the real expression of these activities in a given situation. Here, the
high levels of nitric nitrogen may result from an increase in the numbers of nitrifying bacteria, albeit less active
in the presence of cadmium, coupled with a reduction in the numbers of denitrifying bacteria.
Determination of other activities (dehydrogenase activity, ammonification, ureolysis, proteolysis,
glucosidase, phosphatases . . . . ) and assay of adenyl nucleotides (especially of ATP) will allow a fuller
evaluation of cadmium toxicity in the lagoon pilots.

References
[ 1] O. Ravera, Cadmium in freshwater ecosystems, Experientia, 40, 2-14 (1984).
[2] J.T. Trevors, G.W. Stratton and G.M. Gadd, Cadmium transport, resistance and toxicity in bacteria, algae
and fungi, Can. J. Microbiol., 32,447-464 (1986).
[3] P.T.S. Wong, Toxicity of cadmium to freshwater microorganisms, phytoplankton and invertebrates. In
Cadmium in the Aquatic Environment (Edited by J.O. Nriagu and J.B. Sprague), 117-138, Wiley, New York
(1987).
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