Doklady Biological Sciences, Vol. 380, 2001, pp. 492–495. Translated from Doklady Akademii Nauk, Vol.
380, No. 5, 2001, pp. 714–717.
Original Russian Text Copyright © 2001 by Ostroumov.
GENERAL BIOLOGY
Responses of Unio tumidus to Mixed Chemical Preparations
and the Hazard of Synecological Summation
of Anthropogenic Effects
S. A. Ostroumov
Presented by Academician V.N. Bol’shakov February 26, 2001
Received February 28, 2001
Bivalve mollusks contribute significantly to the
flows of matter and energy in ecosystems [1, 2].
Bivalves were used as a test object in studies on the bio-
logical effects of metals [3], synthetic surfactants, and
other pollutants and xenobiotics on living organisms
[4–10]. Surfactants occur in the environment as compo-
nents of surfactant-containing synthetic washing mix-
tures (SWMs), liquid washing mixtures (LWMs), and
other mixed preparations. Until the present time, there
has been no detailed work aimed at clarifying whether
or not SWMs affect the rate of water filtration by fresh-
water bivalve mollusks.
The goal of this work was to study the effects of
SWMs on the water-filtration activity of the freshwater
bivalve Unio tumidus Philipsson, 1788. The OMO
detergent was used as an example of SWMs.
The mollusks U. tumidus were collected at the par-
tially silted large-grain sand bottom of the Moscow
River upstream from the town of Zvenigorod. Collected
mollusks were kept in tanks with artificial aeration. A
total of 20 mollusk specimens were used in each exper-
iment; 10 specimens were exposed to SWM (variant A)
and the other 10 were control (variant B).
Washing mixtures were added to experimental tanks
5 min before the experiment. The SWM concentrations
shown in Tables 1–3 and indicated in the discussion of
experimental data are the initial concentrations added
to the tanks. In addition to mollusk-containing experi-
mental tanks (variants A and B), control tanks of the
same volume (0.5 l) containing no mollusks were used
to determine the cell suspension density in the absence
of water biofiltration (variants C and D). One of the
control tanks (variant C) contained the same concentra-
tion of SWM as the corresponding experimental tank.
The rate of water biofiltration was estimated as a
decrease in the optical density of the incubation
medium caused by cell elimination from water and the
corresponding decrease in the cell count in water. A
Moscow State University, Vorob’evy gory, Moscow,
119899 Russia
00492$25.00 © 2001 MAIK “Nauka /Interperiodica”
0012-4966/01/0910-
yeast (Saccharomyces cerevisiae) cell suspension was
used. The S. cerevisiae strains MSU1, MSU2, and
MSU3 were from the collection of microorganisms of
the Faculty of Biology, Moscow State University. Yeast
cells were grown for 96 h at 26–28°C in glass flasks on
a flask-shaker (180–200 rpm) under continuous intense
aeration. The culture medium contained 5.0 g/l
(NH4)2SO4, 1.2 g/l KH2PO4, 0.15 g/l KCl, 0.2 g/l
MgSO4 · 7H2O, 0.05 g/l CaCl2, and 50 ml of yeast
autolysate (initial pH, 6.0) [11]. The culture medium
was poured into 0.75-l flasks (150 ml per flask) and
sterilized. A sterile glucose solution was added to the
flasks before inoculation (final concentration, 2 wt %).
The inoculate volume was 3–4 ml per flask. The inocu-
late was preliminarily grown for two days at 30°ë on
wort agar as a slant culture. The biomass was washed
off the agar slant with sterile water and used as an inoc-
ulate. The yeast biomass was provided by N.N. Kolo-
tilova. The optical density at 500 nm was measured
using a Hitachi 200–20 spectrophotometer (Japan) at
an optical path length of 10 mm. The Intelligent
(Automat) OMO detergent (Unilever Polska SA, Byd-
goszcz, Poland) was used as a test surfactant.
Two series of experiments with different strains of
S. cerevisiae were performed. The S. cerevisiae strains
MSU1 and MSU2 were used in experimental series 1
and 2, respectively. The S. cerevisiae cell count was the
same in the two series (7.5 × 106 per ml). The mean
fresh weight of mollusks with shells in variants A and
B of experimental series 1 was 19.2 and 19.0 g, respec-
tively. The mean fresh weight of mollusks with shells in
variants A and B of experimental series 2 was 19.16 and
18.99 g, respectively. The water temperature in experi-
mental series 1 and 2 was 19 and 19.5°C, respectively.
It was shown in our experiments that, in the pres-
ence of the initial SWM concentration of 100 mg/l,
there was a significant decrease in the rate of water fil-
tration by U. tumidus (Table 1). For example, during
1 h of filtration, the optical density of the incubation
medium in experimental (in the presence of SWM,
variant A) and control (variant B) samples decreased to
0.554 and 0.295, respectively. Therefore, the optical
 RESPONSES OF UNIO TUMIDUS TO MIXED CHEMICAL PREPARATIONS 493

Table 1. Dynamics of optical density of the incubation medium in the studies on the effects of the SWM OMO (100 mg/l)
on the filtration of S. cerevisiae MSU1 cells by mollusks Unio tumidus (experiment 1)
Experiment Control Control (without Control (without
Time after addition
(+SWM; +mollusks) (–SWM; +mollusks) mollusks +SWM) mollusks –SWM) A/B, %
of cells, min
(Variant A) (Variant B) (Variant C) (Variant D)
10 0.670 0.660 0.713 0.718 101.5
20 0.667 0.585 0.715 0.706 114.0
30 0.644 0.525 0.676 0.677 122.7
40 0.653 0.442 0.666 0.618 147.7
60 0.554 0.295 0.598 0.587 187.8
70 0.566 0.242 0.581 0.442 233.9

Table 2. Dynamics of optical density of the incubation medium in the studies on the effects of the SWM OMO (50 mg/l) on
the filtration of S. cerevisiae MSU2 cells by Unio tumidus mollusks (experiment 2)

Experiment Control Control (without Control (without

Time after addition
(+SWM; +mollusks) (–SWM; +mollusks) mollusks +SWM) mollusks –SWM) A/B, %
of cells, min
(Variant A) (Variant B) (Variant C) (Variant D)

5 0.711 0.794 0.693 0.671 89.5

20 0.644 0.556 0.702 0.671 115.8
35 0.614 0.448 0.648 0.660 137.1
60 0.557 0.373 0.655 0.629 149.3

density of the experimental sample (in the presence of
SWM) was significantly (187.8%) higher than this
value in the control. The filtration-induced decrease in
the optical density (0.716 units of optical density) dur-
ing 70 min of incubation was (0.716–0.242 =
0.474 units of optical density) and (0.716–0.566 =
0.150 units of optical density) in the control and exper-
imental samples, respectively. Therefore, the rate of the
filtration-induced decrease in the optical density of the
control samples was significantly (316%) higher than
in the experimental samples. This effect can be attrib-
uted to the SWM-induced inhibition of the elimination
of unicellular organisms from water by mollusks. It
also follows from Table 1 that, in the absence of mol-
lusks (i.e., in the absence of water filtration; variants C
and D) the optical density of the incubation medium
was significantly higher than in the control variant with
mollusks and without SWM (variant B).
Similar results were obtained with the cell suspen-
sion of the other strain of S. cerevisiae. It follows from
Table 2 that incubation with SWM (50 mg/l) caused a
decrease in the rate of water filtration. The rate of elim-
ination of unicellular organisms from water by mol-
lusks in the presence of SWM (variant A) was lower
than in the control (variant B). The optical density of
the incubation medium at the end of the experiment
(after 60 min of incubation) in the experimental sam-
ples in the presence of SWM (variant A) was signifi-
cantly (149.3%) higher than the optical density of the
control samples (variant B).
Similar results were obtained with the cell suspen-
sion of the third strain of S. cerevisiae.
In addition, it was found in experiment 2 (S. cerevi-
siae strain MSU2) that, in contrast to variant A (in the
presence of SWM), pellets were excreted by mollusks
after 20 min of incubation of the control samples in
variant B (in the absence of SWM). Although pellets
were also excreted by mollusks in variant A, they
appeared significantly later (after 3 h of incubation) and
their amount was significantly smaller than in variant B.
Therefore, the pellet excretion by mollusks was inhib-
ited in the presence of SWM (variant A).
Thus, the results of our experiments showed that the
synthetic detergent OMO tested in this work caused a
decrease in the rate of water filtration by freshwater
mollusks. In other words, this detergent inhibits the fil-
tration activity of mollusks and decreases the efficiency
of elimination of suspended particles from water. This
conclusion is consistent with experimental results
obtained on marine mollusks. For example, it was
found in our experiments [4–7] that SWMs and LWMs
of various types were capable of inhibiting the water fil-
tration by marine mollusks (Table 3). Therefore, certain
concentrations of the mixed detergents (SWMs and
LWMs) reduce the filtration activity of many species of
bivalve mollusks (water filtrators). Losk-Universal,

DOKLADY BIOLOGICAL SCIENCES Vol. 380 2001

494
Table 3. Inhibition of the filtration activity of organisms limiting the population size of phytoplankton by certain chemical
substances
Chemical substance Concentration, mg/l
SWM OMO 50–100
SWM Losk-Universal 7–20
SWM Tide-Lemon 50
SWM IXI 10–50
SWM Deni-Automat 30
SWM Lanza 20
SWM Vesna-Delikat 1 Crassostrea gigas
LWM E 2 Mytilus galloprovincialis
LWM E 2 Crassostrea gigas
Triton X-100 0.5
Note: (+) effect of inhibition of elimination of unicellular organisms during filtration is observed; (–) lack of statistically significant (95%)
effect of inhibition.
Deni-Automat, Vesna-Delikat, Tide-Lemon, Lanza,
LWM E, and some other mixed detergents also exert
similar effects.
In addition to synthetic surfactants, many SWMs
and LWMs also contain phosphorus compounds. The
content of sodium tripolyphosphate in widely used
detergents such as Bio-S, Kristall, Kashtan, and many
other reaches 3–40 wt %. Phosphates stimulate phy-
toplankton growth. At certain concentrations of SWMs,
the phosphate-induced stimulation of phytoplankton
growth may exceed the potential inhibition effect of the
surfactant [12–15]. Therefore, mixed detergents exert
two types of hazardous effects on ecosystems: phos-
phate-induced stimulation of phytoplankton growth
and surfactant-induced inhibition of biofiltrators.
Because biofiltrators are an effective natural factor of
unicellular plankton population control, the two types
of the detergent-induced effects on an ecosystem facil-
itate the phytoplankton population growth. Therefore,
these effects are summed, thereby increasing the hazard
of the ecosystem collapse.
Thus, the results obtained in this work are of partic-
ular interest in the context of discussion of the potential
ecological danger of pollutants for integral functions of
ecosystems. Synecological summation of the effects of
anthropogenic factors on plankton populations and bio-
filtrators is of particular concern. Therefore, ecological
situations should be analyzed based on the synecologi-
cal approach to the problem. In other words, the inter-
action between populations of plankton organisms and
biofiltrator-consumers should be taken into consider-
ation in studies on the ecological effects of SWMs and
LWMs on these populations. The results obtained in
this work are consistent with the conclusions drawn in
the preceding works [7–9].
OSTROUMOV
Effect of
Organisms References
organic substance
Unio tumidus + This work
Mytilus galloprovincialis + [9]
Mytilus galloprovincialis + The same
Mytilus galloprovincialis + [6]
Crassostrea gigas + The same
Crassostrea gigas + "
+ "
+ "
+ "
Mytilus edulis –(Exposure time, "
30–60 min)
ACKNOWLEDGMENTS
I am grateful to Prof. J. Widdows, and to
V.D. Fedorov, V.V. Malakhov, A.G. Dmitrieva, and
other researchers at Moscow State University, as well
as Yu.Yu. Dgebuadze, N.S. Zhmur, and Yu.P. Kozlov,
for stimulating discussion. I am also grateful to
E.A. Kuznetsov, N.N. Kolotilova, and O.S. Ostroumov
for their expert assistance in some experiments.
This study was supported by IBG and the Open
Society Foundation (RSS project no. 1306/1999) and
partly supported by the MacArthur Foundation (Pro-
gram for Global Safety and Stable Development).
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