Chemosphere 55 (2004) 257–265

www.elsevier.com/locate/chemosphere

Studies on lethal concentrations and toxicity stress

of some xenobiotics on aquatic organisms
M. Abul Farah a, Bushra Ateeq a, M. Niamat Ali b, Rubina Sabir a,
Waseem Ahmad a,*
a
Gene-Tox Laboratory, Division of Genetics, Department of Zoology, Aligarh Muslim University, Aligarh 202002, UP, India
b
P.G. Department of Zoology, University of Kashmir, Srinagar-190006, J & K, India
Received 22 January 2003; received in revised form 3 October 2003; accepted 25 October 2003

Abstract

Three widely used xenobiotics pentachlorophenol (PCP), 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-chloro-2,6-
diethyl-N-(butoxymethyl) acetanilide (Butachlor) are evaluated for acute toxicity and stress behavior on freshwater fish
(Heteropneustes fossilis, Clarias batrachus, Channa punctatus) and mosquito larvae (Culex pipiens fatigans). The
experiment was carried out by medium treatment using intermittent flow-through system. Median lethal concentrations
(LC50 ) were calculated by probit analysis. The LC50 values and 95% confidence intervals showed variable range for
tested chemicals. Mosquito larvae generally appeared resistant than fish, while H. fossilis was found to be most sen-
sitive. Stress signs in the form of behavioral changes are also observed. Both types of organisms are recommended as
good bioindicator for the risk assessment of aquatic environment due to chemicals tested.
Ó 2003 Elsevier Ltd. All rights reserved.

Keywords: Acute toxicity; LC50 ; Behavioral changes; PCP; 2,4-D; Butachlor

1. Introduction (Seiler, 1991; Extoxnet, 1996). The other related com-

Chemicals play an important role in modern day
farm practices. New formulations are introduced on
regular basis. Human destructive influence on the
aquatic environment in the form of sub-lethal pollution
also contributes to chronic stress conditions. Impor-
tant compounds such as (PCP), 2,4-D and Butachlor
belonging to chlorophenoxy and acetanilide groups merit
immediate attention in this context. Together these con-
stitute the highest production figure worldwide (Seiler,
1991; Hill et al., 1997; 2,4-D Fact Sheet PAN, 1997).
The pronounced biocidal activity of PCP, known for
many years, has facilitated its use as fungicide, mollus-
cide, insecticide, herbicide, slimicide and as preservative
*
Corresponding author. Tel.: +91-571-2720920; fax: +91-
571-2708336.
E-mail address: faridi2001@rediffmail.com (W. Ahmad).
0045-6535/$ - see front matter Ó 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/j.chemosphere.2003.10.063
pound, 2,4-D, is a common herbicide employed in post-
emergence foliar spray and has additional use in weed
control of wheat, rice, maize and aquatic weeds (2,4-D
Fact Sheet PAN, 1997), while, Butachlor is known for
its pre-emergence selective herbicidal activity and is used
in rice fields of many Asian countries (Sinha et al., 1995).
The massive use of these chemicals has therefore, re-
sulted in their ubiquitous presence in the environment
and more seriously, exposure of people has culminated
in serious toxic symptoms (Klemmer et al., 1980). De-
spite this, no satisfactory information is available on the
toxic stress of these chemicals with special reference to
India.
For such purpose, initial toxicity tests are usually
performed on water and sediments using fish and
invertebrates. Thus standard lethal tests such as the
median lethal concentration (LC50 , 96 h) are recom-
mended for preliminary exploration to establish initial
estimates of the toxicity of a substance (Sprague, 1989).
258 M.A. Farah et al. / Chemosphere 55 (2004) 257–265
The effects of pollutants on fish can be evaluated by
acute and chronic toxicity test (Sprague, 1973; Rand and
Petrocelli, 1985; Van der Merwe et al., 1993; Nussey
et al., 1996). In addition, abnormal behavior if any is
also recommended as the sign of toxicity stress (EPA
Guideline, 1996).
A few studies on median lethal dose of forestated
chemicals have appeared in terrestrial vertebrates: for
example, 50–328 mg/kg for PCP (RTECS, 1980; Renner
et al., 1986), 100–2000 mg/kg for 2,4-D (Gehring and
Betso, 1977; Young, 2000) and 2000 mg/kg for Buta-
chlor (Tomlin, 1994). The situation however, in aquatic
organisms is complicated since completely or partially
dissolved chemicals maintain a constant pressure. PCP,
2,4-D and Butachlor used terrestrially end up in the
water bodies and create such pressure, affecting non-
target aquatic organisms.
This study is aimed at examining the toxicity of these
compounds on some freshwater fish (Heteropneustes
fossilis, Clarias batrachus and Channa punctatus) and the
larval stages of the mosquito (Culex pipiens fatigans) by
determining LC50 values and analysing behavioral
changes due to toxic stress. The study further focussed
on more reliable evaluation of the risk to freshwater
organisms and to strengthen the base line data that
could be used to estimate a comparative sensitivity to
these xenobiotics.
2. Materials and methods
2.1. Chemicals
The herbicides, pentachlorophenol (CAS No: 87-86-
5, 99% purity), 2,4-dichlorophenoxyacetic acid (CAS
No: 94-75-7, 98% purity) and 2-chloro-2,6-diethyl-N-
(butoxymethyl) acetanilide, 50% EC (Butachlor: CAS
No: 23184-66-9, technical grade) were purchased from
Fluka Chemika (Switzerland), Loba Chemie and Shree
Rasayan Udyog (India) respectively.
2.2. Experimental organisms
Fish: Various freshwater fish: H. fossilis (30 ± 5 g and
18 ± 2 cm), C. batrachus (35 ± 5 g, 16 ± 2 cm) and C.
punctatus (30 ± 5 g, 15 ± 2 cm) were procured from local
out let and acclimatized for two weeks to the laboratory
conditions. All the stocks were maintained in polypro-
pylene troughs containing water at 22 ± 2 °C and the
dissolved oxygen at the constant (5 ppm) level. Specimen
were subjected to a prophylactic treatment by bathing
twice in 0.01% KMnO4 for 30 min at an interval of 24 h
before commencing the routine experiments during
which feeding was stopped. Only healthy fish showing
little or no mortality for two weeks were used in the
experiments.
Mosquito larvae: Fourth instar larval stages of Culex
pipiens fatigans with uniform size were used. All the
stocks were maintained at constant conditions of tem-
perature (25 ± 2 °C) and humidity (80 ± 5%).
2.3. Treatment
Since 2,4-D and PCP are not soluble or have low
solubility, a stock solution of the two chemicals was
prepared by dissolving desired amount in ethanol. The
multiple exposures comprised of 10 concentrations in
each chemical (Tables 1 and 2). Replicates on negative
control (water) and solvent control (0.2% ethanol) were
set up simultaneously for comparison. The acute toxicity
test was worked out in medium treatment by intermit-
tent flow-through test by replacing toxicant and the
dilution of water at regular intervals (Adams, 1993). A
group of 10 fish and 100 larvae were exposed in each
concentration. Treatments were given in polypropylene
troughs by dissolving desired quantity of stock solution
in 50 and 10 l of water for fish and mosquito larvae
respectively. Remaining test conditions like temperature
(22 ± 2 °C), pH of water (6.5–8.5) dissolved oxygen
>60% of saturation and hardness (140–160 mg/l as
CaCO3 ), etc. were kept constant. All variables and test
concentrations were measured at the beginning and the
termination point of the study.
2.4. Determination of LC50 and toxicity stress
Mortality of test organisms was recorded when
opercular movements stopped. Dead individuals were
removed instantly. Accurate records of mortality counts
were maintained at a regular interval of 6 h till 96 h (fish)
and 48 h (C. p. fatigans). LC50 values were based on the
cumulative mortality observed at the end of a desired
exposure. LC50 values and 95% confidence intervals (CI)
were calculated by probit analysis transformation
method and by plotting graph of percent mortality
(probit value) against log concentrations (Musch, 1996).
Change in behavior related to stress and symptoms were
closely monitored during the course of experiment.
3. Results
3.1. LC50 determination
The relation between various concentrations of PCP,
2,4-D and Butachlor and the percent mortality with
their respective probit values for the test organisms are
shown in Tables 1 and 2. The estimated LC50 values and
confidence intervals are listed in Table 3. Graphic rep-
resentation of the probit transformation for the chemi-
cals under study are presented in Figs. 1–3. No mortality
Table 1
Showing the percent mortality and their respective probit values obtained by acute toxicity testing by various concentrations of PCP, 2,4-D and Butachlor in H. fossilis and
C. batrachus
Test PCP 2,4-D Butachlor
organism Conc. Log % Mortality Probit Conc. Log % Mortality Probit Conc. Log % Mortality Probit

M.A. Farah et al. / Chemosphere 55 (2004) 257–265

ppm conc. value ppm conc. value ppm conc. value
H. fossilis 0.35 )0.52 0 – 30 1.47 0 – 0.5 )0.30 0 –
0.40 )0.45 3.34 3.12 40 1.60 6.67 3.5 1.0 0 16.67 4.05
0.45 )0.39 10 3.72 50 1.69 23.34 4.26 1.5 0.17 20 4.16
0.50 )0.34 16.67 4.05 60 1.77 23.34 4.26 2.0 0.30 40 4.75
0.55 )0.30 33.34 4.56 70 1.84 36.67 4.67 2.5 0.39 53.34 5.08
0.60 )0.25 43.34 4.82 80 1.90 46.67 4.92 3.0 0.47 63.34 5.33
0.65 )0.22 53.34 5.08 90 1.95 60 5.25 3.5 0.54 73.34 5.61
0.70 )0.18 63.34 5.33 100 2 73.34 5.61 4.0 0.60 80 5.84
0.75 )0.15 80 5.84 110 2.04 83.34 5.95 4.5 0.65 90 6.28
0.80 )0.12 100 – 120 2.07 100 – 5.0 0.69 100 –

C. batrachus 0.35 )0.45 0 – 70 1.84 0 – 1.0 0 0 –

0.40 )0.39 3.34 3.12 80 1.90 6.67 3.52 1.5 0.17 13.34 3.87
0.45 )0.34 10 3.72 90 1.95 10 3.72 2.0 0.30 26.67 4.39
0.50 )0.30 23.34 4.26 100 2 13.34 3.87 2.5 0.39 33.34 4.56
0.55 )0.25 36.67 4.67 110 2.04 26.67 4.39 3.0 0.47 43.34 4.82
0.60 )0.22 46.67 4.92 120 2.07 43.34 4.82 3.5 0.54 53.34 5.08
0.65 )0.18 56.67 5.18 130 2.11 63.34 5.33 4.0 0.60 60 5.25
0.70 )0.15 60 5.25 140 2.14 70 5.70 4.5 0.65 80 5.84
0.75 )0.12 76.67 5.74 150 2.17 83.34 5.90 5.0 0.69 83.34 5.95
0.80 )0.09 100 – 160 2.20 100 – 5.5 0.74 100 –

259
260 M.A. Farah et al. / Chemosphere 55 (2004) 257–265

Table 2
Showing the percent mortality and their respective probit values obtained by acute toxicity testing by various concentrations of PCP,
2,4-D and Butachlor in C. punctatus and C. p. fatigans
Test PCP 2,4-D Butachlor
organism Conc. Log % Probit Conc. Log % Probit Conc. Log % Probit
ppm conc. Mortality value ppm conc. Mortality value ppm conc. Mortality value
C. punctatus 0.50 )0.30 0 – 60 1.77 0 – 0.5 )0.30 0 –
0.55 )0.25 6.67 3.52 70 1.84 10 3.72 1.0 0 13.34 3.87
0.60 )0.22 16.67 4.05 80 1.90 16.67 4.05 1.5 0.17 26.67 4.39
0.65 )0.18 23.34 4.26 90 1.95 33.34 4.50 2.0 0.30 33.34 4.56
0.70 )0.15 36.67 4.39 100 2 40 4.75 2.5 0.39 43.34 4.82
0.75 )0.12 46.67 4.92 110 2.04 56.67 5.18 3.0 0.47 52 5.05
0.80 )0.09 53.34 5.08 120 2.07 73.34 5.61 3.5 0.54 56.67 5.18
0.85 )0.07 66.67 5.44 130 2.11 80 5.84 4.0 0.60 73.34 5.61
0.90 )0.04 80 5.84 140 2.14 86.67 6.13 4.5 0.65 80 5.84
0.95 )0.02 100 – 150 2.17 100 – 5.0 0.69 100 –

C. p. fatigans 20 1.30 0 – 50 1.69 0 – 5 0.69 0 –

40 1.60 21 4.19 100 2 7 3.52 10 1 20 4.16
60 1.77 25 4.33 150 2.17 15 3.96 20 1.30 37 4.67
80 1.90 25 4.33 200 2.30 25 4.33 30 1.47 43 4.82
100 2 51 5.03 250 2.39 39 4.72 40 1.60 52 5.05
120 2.07 64 5.36 300 2.47 48 4.95 50 1.69 63 5.33
140 2.14 75 5.67 350 2.54 57 5.18 60 1.77 75 5.67
160 2.20 80 5.84 400 2.60 71 5.55 70 1.84 83 5.95
180 2.25 87 6.13 450 2.65 84 5.99 80 1.90 94 6.55
200 2.30 100 – 500 2.69 100 – 90 1.95 100 –

Table 3
Calculated LC50 values (ppm) and 95% confidence intervals for PCP, 2,4-D and Butachlor in fish and mosquito larvae
Test organism Durations (h) LC50 (95% confidence intervals)
PCP 2,4-D Butachlor
H. fossilis 96 0.58 (0.40–0.70) 81 (50–100) 2.34 (1.5–4.0)
C. batrachus 96 0.64 (0.45–0.75) 122 (100–130) 3.25 (2.5–4.5)
C. punctatus 96 0.77 (0.65–0.90) 107 (90–130) 2.82 (2.0–4.5)
C. p. fatigans 48 98 (60–140) 302 (150–400) 35 (20–70)

was observed in any fish or in mosquito larvae in control
group.
The LC50 values estimated in H. fossilis, C. batrachus,
C. punctatus and C. p. fatigans were 0.58, 0.64, 0.77 and
98 ppm for PCP, 81, 122, 107 and 302 ppm for 2,4-D
and 2.34, 3.25, 2.82 and 35 ppm for Butachlor respec-
tively. Concentrations below 0.35 ppm (PCP), 30 ppm
(2,4-D) and 0.5 ppm (Butachlor) showed no mortality in
any fish, whereas, no mortality appeared in mosquito
larvae at or below 20, 50 and 5 ppm of the respective
chemicals. In terms of relative sensitivity, H. fossilis was
found to be more sensitive among the fish and mosquito
larvae comparatively resistant than the fish, as the esti-
mated LC50 values were higher for each chemical.
3.2. Toxic stress and poisoning symptoms in fish
Fish subjected to higher concentrations of chemicals
under observation displayed behavioral changes such
as restlessness, swimming at the surface or abnormal
swimming behavior, vigorous jerks of body, loss of
balance, myotonia and anorexia. The concentrations
between 0.65–0.90 ppm, 110–150 ppm and 4.0–5.0 ppm
of PCP, 2,4-D and Butachlor induced lethality accom-
panied by tremors and coma. Severe poisoning was
found to be terminated into death along with the
observable signs of congestion, breathing difficulties,
edema and hemorrhages of the viscera.
3.3. Toxic stress and poisoning symptoms in mosquito
larvae
Behavioral and the swimming patterns were normal
in control group without any mortality. Concentrations
>LC50 that is 120–200 ppm (PCP), 350–500 ppm (2,4-D)
and 50–90 ppm (Butachlor) showed stress in larval
stages. Respiratory problem was observed in most cases
forcing the larvae to float on the surface in congrega-
 M.A. Farah et al. / Chemosphere 55 (2004) 257–265 261

100 10 100 10
H. fossilis C. batrachus
90 9 90 9
80 8 80 8
70 7 70 7

Probit Value

Probit Value
% Mortality
% Mortality

60 6 60 6
50 5 50 5
40 4 40 4
30 3 30 3
20 2 20 2
10 1 10 1
0 0 0 0
-0.52 -0.45 -0.39 -0.34 -0.3 -0.25 -0.22 -0.18 -0.15 -0.12 -0.45 -0.39 -0.34 -0.3 -0.25 -0.22 -0.18 -0.15 -0.12 -0.09
Log Concentrations PCP (ppm) Log Concentrations PCP (ppm)

100 10
C. p. fatigans
100 10 90 9
C. punctatus
90 9 80 8
80 8 70 7
70 7

Probit Value
% Mortality
60 6
Probit Value
% Mortality

60 6
50 5
50 5
40 4 40 4

30 3 30 3
20 2 20 2
10 1 10 1
0 0
0 0
-0.3 -0.35 -0.22 -0.18 -0.15 -0.12 -0.09 -0.07 -0.04 -0.02
1.3 1.6 1.77 1.9 2 2.07 2.14 2.2 2.25 2.3
Log Concentrations PCP (ppm) Log Concentrations PCP (ppm)

Fig. 1. Graphic representation of log concentrations versus the probit values of the percent mortality determining LC50 of PCP in the
organisms tested.

tional form. Affected larvae gradually become inacti- in cutthroat trout depending on the formulations used
vated avoiding vertical movements, followed by death (NRCC, 1978). Other LC50 values reported are 1.1 mg/l
as a consequent. for rainbow trout (Tomlin, 1994) and 63.24 mg/l for C.
carpio (Sarikaya and Yilmaz, 2003). The corresponding
LC50 values for the present, range from 81 to 122 ppm
4. Discussion in all the fish studied. LC50 (96-h) values of Butachlor
has been reported by Tomlin (1994). These are 0.52 mg/l
Toxicity of chemicals to aquatic organisms has for rainbow trout, 0.44 mg/l for bluegill sunfish, 0.32
shown to be effected by age, size and health of the spe- mg/l for carp and 0.14 mg/l for channel catfish, in
cies. Physiological parameters like quality, temperature, comparison present value ranges between 2.34 and 3.25
pH, dissolved oxygen and turbidity of water, amount ppm for this chemical showing significant toxicity to
and kind of aquatic vegetation, concentration and for- the fish under study and possibly to other organisms as
mulation of chemical and its exposure also greatly well.
influence such studies (EPA Pesticide Fact Sheet, 2000; A review of literature revealed little information on
Young, 2000). Johnson and Finley (1980), found PCP as toxicity of these herbicides in invertebrate group using
highly toxic to many species of fish as evident from the mosquito larvae. Aquatic invertebrates do not in
reported 96-h LC50 in Chinook (68 lg/l), rainbow trout general seem to be very sensitive to 2,4-D and the
(52 lg/l), fathead minnow (205 lg/l), channel catfish (68 toxicity level show wide range )0.1 to >100 ppm (2,4-
lg/l) and bluegill sunfish (32 lg/l). Against these, the D Fact Sheet PAN, 1997). The median tolerance limit
present values recorded for the same chemical are 0.58, at 48 h in Culex pipiens by alkyl benzene sulfonate
0.64 and 0.77 ppm in H. fossilis, C. batrachus and C. (detergent) was estimated by Lal et al. (1983). In the
punctatus respectively. present case, LC50 48-h is estimated between 35 and
As for the 2,4-D, some formulations are highly toxic 302 ppm. Toxic effects of heavy metals on Aedes ae-
(esters), therefore, LC50 ranges between 1.0 and 100 mg/l gypti larvae reported LC50 endpoints as 3.1, 16.5 and
262 M.A. Farah et al. / Chemosphere 55 (2004) 257–265

100 10 100 10
H. fossilis C. batrachus
90 9 90 9
80 8 80 8
70 7 70 7

Probit Value

Probit Value
% Mortality
% Mortality

60 6 60 6
50 5 50 5
40 4 40 4
30 3 30 3

20 2 20 2

10 1 10 1
0 0 0 0
1.47 1.6 1.69 1.77 1.84 1.9 1.95 2 2.04 2.07 1.84 1.9 1.95 2 2.04 2.07 2.11 2.14 2.17 2.2

Log Concentrations 2,4-D (ppm) Log Concentrations 2,4-D (ppm)

100 10
100 10 C. p. fatigans
C. punctatus 90 9
90 9
80 8
80 8
70 7
70 7 Probit Value

Probit Value
60 6
60 6 % Mortality
% Mortality

5 50 5
50
4 40 4
40
30 3 30 3

20 2 20 2

10 1 10 1

0 0 0 0
1.77 1.84 1.9 1.95 2 2.04 2.07 2.11 2.14 2.17 1.69 2 2.17 2.3 2.39 2.47 2.54 2.6 2.65 2.69

Log Concentrations 2,4-D (ppm) Log Concnetrations 2,4-D (ppm)

Fig. 2. Graphic representation of log concentrations versus the probit values of the percent mortality determining LC50 of 2,4-D in the
organisms tested.

33 ppm for Hg, Cd and Cu respectively (Ryms-Keller fauna like snails, slug worms, tadpoles and fish finger-
et al., 1998). lings.
Other chemicals like carbaryl, malathion and Behavioral changes as a result of stress are further
ammonium sulfate have also been tried on H. fossilis accepted as the most sensitive indication of potential
(Singh et al., 1984; Banerjee, 1993). LC50 value of car- toxic effects. Various behavioral changes observed dur-
baryl (15.08 mg/l, 96 h) in C. batrachus induces pertur- ing the coarse of the study are restlessness, swimming at
bations at the level of certain biochemical components the surface or abnormal swimming behavior, vigorous
(Sharma, 1999). Similarly few acute toxicity studies in jerks of body, loss of balance, myotonia and anorexia.
Channa punctatus are carried out and LC50 values cal- More or less similar changes were reported in common
culated for malathion (4.51 mg/l, 96-h), endosulfan (5.78 carp (C. carpio) (Sarikaya and Yilmaz, 2003) and gup-
mg/l, 96-h), carbaryl (8.71 ppm, 48-h) and phenthoate pies (Peocilia reticulata) by Polat et al. (2002) and Baser
(0.473 ppm, 48-h) (Rao et al., 1985; Haider and Inbaraj, et al. (2003) using various chemicals including 2,4-D.
1986). Ferrando et al. (1991), in their study on the effect of
From a comparative account, H. fossilis seems to be eight selected organochlorine pesticides on eels (Anguilla
more sensitive among the fish examined (C. batrachus anguilla), while determining 96-h LC50 values also re-
and C. punctatus) as the LC50 values are found to be ported behavioral changes like anxiety, disorders in
lower for all the tested chemicals. Comparatively, swimming patterns, loss of balance excessive mucous
mosquito larvae appear generally resistant and their secretion and lightening in colour. Sub-lethal concen-
LC50 values are higher than those in the fish. This trations of carbaryl in catfish Mystus vittatus, acceler-
suggests that these chemicals have greater margin of ated the swimming activity and increased the frequency
safety to mosquito larvae in comparison to fish. In a of opercular beats (Arunachalam et al., 1980). In other
similar study by Lal et al. (1983), Culex pipiens was aquatic species including mosquito larvae (Culex pipi-
found to be more resistant than other species of aquatic ens) behavioral changes as an index of detergent toxicity
 M.A. Farah et al. / Chemosphere 55 (2004) 257–265 263

100 10 100 10
H. fossilis C. batrachus
90 9 90 9
80 8 80 8
70 7 70 7
60 6

Probit Value
% Mortality

Probit Value
60 6

% Mortality
50 5 50 5

40 4 40 4

30 3 30 3

20 2 20 2

10 1 10 1

0 0 0 0
-0.3 0 0.17 0.3 0.39 0.47 0.54 0.6 0.65 0.69 0 0.17 0.3 0.39 0.47 0.54 0.6 0.65 0.69 0.74

Log Concentrations Butachlor (ppm) Log Concentrations Butachlor (ppm)

100 10 100 10
C. punctatus C. p. fatigans
90 9 90 9

80 8 80 8

70 7 70 7
Probit Value

Probit Value
% Mortality
% Mortality

60 6 60 6

50 5 50 5

40 4 40 4

30 3 30 3

20 2 20 2
10 1 10 1
0 0 0 0
-0.3 0 0.17 0.3 0.39 0.47 0.54 0.6 0.65 0.69 0.69 1 1.3 1.47 1.6 1.69 1.77 1.84 1.9 1.95
Log Concentrations Butachlor (ppm) Log Concentrations Butachlor (ppm)

Fig. 3. Graphic representation of log concentrations versus the probit values of the percent mortality determining LC50 of Butachlor in
the organisms tested.

are known (Lal et al., 1983). The mode of action of all inhibition of cholinesterase (Rajyalakshmi et al., 1996).
these chemicals may be markedly different, behavioral In contrast the respiratory problem in mosquito larvae
changes are similar to ours. could be due to external factors such as water pH,
It is widely accepted that the stress response as a mineral composition and ionic calcium levels (Wend-
whole is characterised by physiological changes. These elaar Bonga, 1997).
changes tend to be similar for stressors and could be The present study on the acute toxicity in aquatic
as varied as anesthesia, flight, forced swimming, disease organisms is expected to help assessment of possible risk
treatments, handling, scale loss, or rapid temperature to similar species in natural environment, and as an aid
change (Wedenmeyer and McLeay, 1981). A study by in determination of water quality criteria for regula-
Wendelaar Bonga (1997) showed that stressor increases tory measures. Correlation with acute toxicity testing
the permeability of the surface epithelia, including the of other species for comparative purpose is further
gills to water and ions and thus induces hydromineral achieved by such studies.
disturbances. Therefore, it is good indicator of toxicity
in fish than in mammals, as the fish are exposed to
aquatic pollutants by extensive and delicate respiratory Acknowledgements
surface of the gills. The high bioavailability of many
chemicals in water is an additional factor in their sen- The Council of Scientific and Industrial Research
sitivity to stress. (CSIR), New Delhi, India is profusely acknowledged for
The study showed that PCP and Butachlor induced financial help no. 9/112 (339) 2002 EMR-I-SRF; 9/112
the stress well below the LC50 concentrations, while in (340) 2002 EMR-I-SRF and 13(7740-A)/2002-pool. The
case of 2,4-D, the stress response appeared at >LC50 Chairman, Department of Zoology, AMU, Aligarh is
concentrations. Stress response by Butachlor was gen- gratefully acknowledged for providing necessary facili-
erally severe with the possible neurotoxic effect through ties.
264 M.A. Farah et al. / Chemosphere 55 (2004) 257–265
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