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

00492$25.00 © 2001 MAIK “Nauka /Interperiodica”
0012-4966/01/0910-
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 cantly (149.3%) higher than the optical density of the
SWM) was significantly (187.8%) higher than this control samples (variant B).
value in the control. The filtration-induced decrease in Similar results were obtained with the cell suspen-
the optical density (0.716 units of optical density) dur- sion of the third strain of S. cerevisiae.
ing 70 min of incubation was (0.716–0.242 =
0.474 units of optical density) and (0.716–0.566 = In addition, it was found in experiment 2 (S. cerevi-
0.150 units of optical density) in the control and exper- siae strain MSU2) that, in contrast to variant A (in the
imental samples, respectively. Therefore, the rate of the presence of SWM), pellets were excreted by mollusks
filtration-induced decrease in the optical density of the after 20 min of incubation of the control samples in
control samples was significantly (316%) higher than variant B (in the absence of SWM). Although pellets
in the experimental samples. This effect can be attrib- were also excreted by mollusks in variant A, they
uted to the SWM-induced inhibition of the elimination appeared significantly later (after 3 h of incubation) and
of unicellular organisms from water by mollusks. It their amount was significantly smaller than in variant B.
also follows from Table 1 that, in the absence of mol- Therefore, the pellet excretion by mollusks was inhib-
lusks (i.e., in the absence of water filtration; variants C ited in the presence of SWM (variant A).
and D) the optical density of the incubation medium Thus, the results of our experiments showed that the
was significantly higher than in the control variant with synthetic detergent OMO tested in this work caused a
mollusks and without SWM (variant B). decrease in the rate of water filtration by freshwater
mollusks. In other words, this detergent inhibits the fil-
Similar results were obtained with the cell suspen- tration activity of mollusks and decreases the efficiency
sion of the other strain of S. cerevisiae. It follows from of elimination of suspended particles from water. This
Table 2 that incubation with SWM (50 mg/l) caused a conclusion is consistent with experimental results
decrease in the rate of water filtration. The rate of elim- obtained on marine mollusks. For example, it was
ination of unicellular organisms from water by mol- found in our experiments [4–7] that SWMs and LWMs
lusks in the presence of SWM (variant A) was lower of various types were capable of inhibiting the water fil-
than in the control (variant B). The optical density of tration by marine mollusks (Table 3). Therefore, certain
the incubation medium at the end of the experiment concentrations of the mixed detergents (SWMs and
(after 60 min of incubation) in the experimental sam- LWMs) reduce the filtration activity of many species of
ples in the presence of SWM (variant A) was signifi- bivalve mollusks (water filtrators). Losk-Universal,

DOKLADY BIOLOGICAL SCIENCES Vol. 380 2001
494 OSTROUMOV

Table 3. Inhibition of the filtration activity of organisms limiting the population size of phytoplankton by certain chemical
substances
Effect of
Chemical substance Concentration, mg/l Organisms References
organic substance
SWM OMO 50–100 Unio tumidus + This work
SWM Losk-Universal 7–20 Mytilus galloprovincialis + [9]
SWM Tide-Lemon 50 Mytilus galloprovincialis + The same
SWM IXI 10–50 Mytilus galloprovincialis + [6]
SWM Deni-Automat 30 Crassostrea gigas + The same
SWM Lanza 20 Crassostrea gigas + "
SWM Vesna-Delikat 1 Crassostrea gigas + "
LWM E 2 Mytilus galloprovincialis + "
LWM E 2 Crassostrea gigas + "
Triton X-100 0.5 Mytilus edulis –(Exposure time, "
30–60 min)
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, ACKNOWLEDGMENTS
LWM E, and some other mixed detergents also exert I am grateful to Prof. J. Widdows, and to
similar effects. V.D. Fedorov, V.V. Malakhov, A.G. Dmitrieva, and
In addition to synthetic surfactants, many SWMs other researchers at Moscow State University, as well
and LWMs also contain phosphorus compounds. The as Yu.Yu. Dgebuadze, N.S. Zhmur, and Yu.P. Kozlov,
content of sodium tripolyphosphate in widely used for stimulating discussion. I am also grateful to
detergents such as Bio-S, Kristall, Kashtan, and many E.A. Kuznetsov, N.N. Kolotilova, and O.S. Ostroumov
other reaches 3–40 wt %. Phosphates stimulate phy- for their expert assistance in some experiments.
toplankton growth. At certain concentrations of SWMs, This study was supported by IBG and the Open
the phosphate-induced stimulation of phytoplankton Society Foundation (RSS project no. 1306/1999) and
growth may exceed the potential inhibition effect of the partly supported by the MacArthur Foundation (Pro-
surfactant [12–15]. Therefore, mixed detergents exert gram for Global Safety and Stable Development).
two types of hazardous effects on ecosystems: phos-
phate-induced stimulation of phytoplankton growth
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DOKLADY BIOLOGICAL SCIENCES Vol. 380 2001