Chemosphere 41 (2000) 177±182

E€ects of 2,4-D, glyphosate and paraquat on growth,

photosynthesis and chlorophyll±a synthesis of Scenedesmus
quadricauda Berb 614
P.K. Wong *

Department of Biology, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong SAR, People's Republic of China

Abstract

The e€ects of 2,4-D, glyphosate and paraquat on growth, photosynthesis and chlorophyll-a synthesis by a freshwater
green alga, Scenedesmus quadricauda Berb 614, were determined. These herbicides are the most often used in Hong
Kong. Within the concentration range 0.02±200 mg/l, paraquat was more toxic than glyphosate and 2,4-D to the
growth, photosynthesis and chlorophyll-a synthesis. The presence of 0.02, 0.2 or 2 mg/l of 2,4-D was not toxic to the
alga. Algal growth, photosynthesis and chlorophyll-a synthesis were stimulated by the presence of low concentrations
(0.02 or 0.2 and 0.02 mg/l, respectively) of 2,4-D and glyphosate. The presence of 0.02 or 0.2 mg/l of paraquat, 2 mg/l of
glyphosate or 20 mg/l of 2,4-D was signi®cantly inhibitory to the three test parameters, whereas the presence of 2 or
more mg/l of paraquat, 20 or more mg/l of glyphosate or 200 mg/l of 2,4-D completely inhibited algal growth, pho-
tosynthesis and chlorophyll-a synthesis. The use of the alga as a bio-indicator of herbicide contamination in freshwater
environment was discussed. Ó 2000 Elsevier Science Ltd. All rights reserved.

1. Introduction Previous studies have indicated that plants and uni-

cellular green algae showed similar responses to the
The pollution of aquatic system by agricultural pol- presence of various chemicals (Sumida and Ueda, 1974;
lutants has attracted great concerns from the public. Muromtzev et al., 1975). There are numerous reports on
Numerous studies have been conducted to determine the the toxicity of individual herbicides in pure form to
harm of these pollutants on living organisms in aquatic di€erent algae (Sullivan et al., 1981; Moorhead and
system (Fargasova, 1994a; Peterson et al., 1994; Fair- Kosinski, 1986; Laurent et al., 1992; Kasai and Hata-
child et al., 1997). Among living aquatic organisms, keyama, 1993; Brown and Lean, 1995; Kobraei and
phytoplankton such as unicellular algae which are im- White, 1996; Okay and Gaines, 1996). Irmer et al. (1986)
portant components of the aquatic food chain, has at- reported the Chlorella sp., the most well-studied uni-
tracted most of the concerns. cellular green alga, was less sensitive than other green
Herbicides are chemicals commonly used to control algae such as Chlamydomonas sp. towards exposure to
weeds in agricultural activities. Some of the applied chemicals; variations in sensitivity of di€erent algal
herbicides are continuously discharged into aquatic en- species to selected toxicants including herbicides have
vironments through the surface runo€, such discharge been frequently reported (Sullivan et al., 1981; Irmer
can lead to contaminated aquatic environments which et al., 1986; Moorhead and Kosinski, 1986; Laurent
are hazardous to resident organisms (Fargasova, 1994b). et al., 1992; Kasai and Hatakeyama, 1993; Brown and
Lean, 1995; Kobraei and White, 1996; Okay and Gaines,
1996). It is very dicult to compare the toxicity of these
herbicides tested by di€erent algal species.
*
Tel.: +852-2609-6383; fax: +852-2603-5767. Numerous studies on toxicity of inorganic and or-
E-mail address: [email protected] (P.K. Wong). ganic compounds to Scenedesmus quadricauda have been

0045-6535/00/$ - see front matter Ó 2000 Elsevier Science Ltd. All rights reserved.
PII: S 0 0 4 5 - 6 5 3 5 ( 9 9 ) 0 0 4 0 8 - 7
178 P.K. Wong / Chemosphere 41 (2000) 177±182

conducted (Ibrahim, 1990; Fargasova, 1993; Hosetti rim, 1997). The herbicides in emulsi®able concentrate
et al., 1993; Saenz et al., 1993; Fargasova, 1994a,b,c; forms were diluted with sterile distilled water and added
Peterson et al., 1994; Reddy and Prasad, 1994; Bussieres into the sterile MBM in various concentrations (in mg/l
et al., 1995; Fargasova and Kizlink, 1996; Tisler and of active ingredient) without sterilization.
Zagorc-Koncan, 1997) since the alga is frequently found For growth inhibition experiments, the procedures of
in freshwater environment and is an important compo- Wong and Chang (1988) were adopted. In brief, 10 ml of
nent in freshwater food chains (Fargasova, 1994c). Only algal suspension from a late log phase culture was in-
a few of these studies concerned the toxicity of herbi- oculated to a 500 ml cotton-plugged Erlenmeyer ¯ask
cides (Ibrahim, 1990; Fargasova, 1994a,c; Peterson with 90 ml of MBM containing various concentrations
et al., 1994). The study of Ibrahim (1990) focused on the of individual herbicide to yield an initial cell density of
e€ect of paraquat on the growth of the alga, while the 5 ´ 104 cells/ml. The algal culture was grown on a gy-
studies of Fargasova (1994a,c) were on the e€ects of rotary shaker running at 100 rpm in a constant tem-
plant growth hormones such as 2,4-D on the algal perature room at 25 ‹ 2°C. Cool white ¯uorescence
growth. Although Peterson et al. (1994) compared the tubes with a light intensity of 80 lE/m2 /s and 16/8 light-
e€ects of 23 pesticides on the 14 C uptake by S. quadric- dark cycle were employed. The growth of the algal cul-
auda. The study included 2,4-D and glyphosate, but not ture was measured by monitoring the changes of cell
paraquat. In the present study, toxicity of three common density by direct cell count with a hematocytometer
but chemically di€erent herbicides, namely, 2,4-D, under a Nikon phase-contrast light microscope at two
glyphosate and paraquat, to the growth, photosynthesis day-intervals until stationary phase was reached. The
and chlorophyll-a synthesis of S. quadricauda Berb 614 growth rate of algal culture was determined by the
were compared. Results obtained from the comparative following equation:
study provide useful information in choosing a suitable
herbicide for agricultural practices base on the e€ects of
these herbicides on the growth and metabolisms of algae Table 1
and plants. Composition of modi®ed bristolÕs medium (MBM)
Chemical Amounts
NaNO3 0.25 g
2. Materials and methods CaCl2 2H2 O 0.025 g
MaSO4 7H2 O 0.075 g
A culture of the freshwater green alga, S. quadricauda K2 HPO4 0.075 g
Berb 614, was originally obtained from University of KH2 PO4 0.17 g
Texas at Austin Culture Collection and kept at the Bi- NaCl 0.025 g
ology Department, The Chinese University of Hong *Trace element (100 X) 10 ml
Kong. The algal cells were grown in Modi®ed BristolÕs Distilled water 990 ml
Medium (MBM, Table 1) and the pH of the medium pH is adjusted to 6.8
*Trace elements (100 X)
was adjusted to 6.8.
EDTANa2 540 mg
Herbicices 2,4-D (2,4-Dichlorophenoxyacetic acid, FeCl3 6H2 O 500 mg
active ingredient 49%) and paraquat (1,10 -dimethyl-4,40 - Heated in 500 ml distilled water until distilled, then added:
bipyridinium ion, active ingredient 20%) manufactured CaCl2 6H2 O 26.55 mg
by Imperial Chemical Industries (ICI), England and Col2 6H2 O 0.02 mg
glyphosate (N-(phosphonomethyl) glycine, active ingre- CuSO4 5H2 O 0.01 mg
dient 35.6%) manufactured by Monsanto, USA were MnCl2 4H2 O 0.3 mg
purchased from commercial sources in Hong Kong. Na2 MoO4 2H2 O 0.05 mg
Chemical, chemical classes, chemical names, chemical ZnSO4 7H2 O 0.04 mg
abstract numbers and modes of action of the herbicides Diluted to 1000 ml with distilled and sterilized by
autoclaving
are listed in Table 2 (Worthing and Walker, 1987; Ka-

Table 2
Chemical formulation and modes of action of herbicides tested with Scenedesmus quadricauda Berb 614 (Karim, 1997)
Herbicide Chemical class Chemical name and number (Chemical abstract service) Mode of action
2,4-D Phenoxy-acetic acid 2,4-Dichlorophenoxyacetic acid; CAS no. 94-75-7 Auxin simulator
Glyphosate Amine N-(phosophonomethyl) glycine; CAS no. 1071-83-6 Inhibit aromatic amino
acids synthesis
Paraquat Pyridine 1,10 -Dimethyl-4,40 -bipyridinium ion; CAS no. 1910-42-5 Alters oxidative
phosphorylation
 P.K. Wong / Chemosphere 41 (2000) 177±182 179

k ˆ … ln X1 ÿ ln X0 †=…T1 ÿ T0 †; Table 3
Growth rate of Scenedesmus quadricauda Berb 614 in the
where k is the speci®c growth rate, X1 the cell density at presence of various concentrations of 2,4-D, glyphosate and
time T1 and X0 is the cell density at time T0 . All ex- paraquat
perimental cultures, including the control, were set up in Herbicide Concentration Growth rateA
triplicate. (mg/l) (k/h)
Gross photosynthetic rate was measured with a Control ) 0.028 ‹ 0.002a B
Gilson di€erential respirometer (Gilson Medical Elec- 2,4-D 0.02 0.034 ‹ 0.004b
tronic, Middleton, Wisconsin, USA). Algal density in 0.2 0.032 ‹ 0.003b
each reaction ¯ask was about 1 ´ 106 cells/ml and the 2 0.028 ‹ 0.003a
algal suspension was placed in main chamber of the 20 0.009 ‹ 0.001c
reaction ¯ask. Appropriate concentrations of herbicide 200 NGdC
and phosphate bu€er (pH 6.8) containing 50 mm of Glyphosate 0.02 0.033 ‹ 0.002b
NaHCO3 which provided carbonate ions for photosyn- 0.2 0.028 ‹ 0.003b
2 0.010 ‹ 0.001c
thesis were placed in the side arms of the reaction ¯asks.
20 NGd
The central well contained 0.2 ml of 10% KOH solution. 200 NGd
The reaction ¯asks were equilibriated at 25°C for 10 min Paraquat 0.02 0.015 ‹ 0.002e
before mixing the herbicide and the algal suspensions in 0.2 0.004 ‹ 0.004f
the main chambers. Photosynthesis activities of control 2 NGd
and treatment ¯asks were measured by recording the 20 NGd
readings of micrometers in 10 min intervals for 60 min 200 NGd
after mixing the herbicide with algal suspension. All A
Mean value ‹ standard deviation of triplicates.
experimental cultures, including the control, were set up B
Di€erence between means of various concentrations of test
in triplicate. herbicide on algal growth by one-way ANOVA followed by
Chlorophyll-a of the algal culture at log and sta- DuncanÕs mutliple range test at P < 0.05. Control and treat-
tionary phases of growth were determined by the ments, statistically identical, are marked by the same letter.
C
method of Jensen (1978). Triplicates were set up for each NG: no growth.
test and control experiments.
Data were analyzed by a computer software, Simg-
aStat version 1.0 (Jandel Scienti®c Softwares, 1995).
Di€erence between individual treatments and control Table 4
Photosynthetic rate of Scenedesmus quadricauda Berb 614 in the
were analyzed using Analysis of Variance (ANOVA)
presence of various concentrations of 2,4-D, glyphosate and
followed by DuncanÕs Multiple Range Test (P < 0.05) paraquat
(Wong and Chang, 1991).
Herbicide Concentration Photosynthetic
(mg/l) rateA (ll O2 /ml/h)
Control ) 128.5 ‹ 4.2aB
3. Results 2,4-D 0.02 135.2 ‹ 6.1b
0.2 132.1 ‹ 4.3b
The presence of low concentration (i.e., 0.02 or 0.2 2 102.5 ‹ 3.2c
mg/l) of 2,4-D or (0.02 mg/l) glyphosate stimulated the 20 75.4 ‹ 4.4d
growth rate of S. quadricauda Berb 614 and the presence 200 NDeC
of 0.2 mg/l of glyphosate and 2 mg/l of 2,4-D did not Glyohosate 0.02 134.7 ‹ 5.6b
inhibit algal growth (Table 3). The presence of 0.02 or 0.2 127.9 ‹ 3.2a
0.2 mg/l of paraquat, 2 mg/l of glyphosate and 20 mg/l of 2 80.2 ‹ 2.6d
2,4-D signi®cantly inhibited algal growth, while the 20 NDe
200 NDe
presence of 2 mg/l or more of paraquat, 20 mg/l or more
Paraquat 0.02 108.4 ‹ 4.8c
of glyphosate and 200 mg/l of 2,4-D completely inhib- 0.2 78.9 ‹ 2.5d
ited algal growth (Table 3). 2 NDe
Similar to the e€ects of the herbicides to algal 20 NDe
growth, the presence of low concentration (i.e., 0.02 or 200 NDe
0.2 mg/l) of 2,4-D or (0.02 mg/l) of glyphosate increased A
Mean value ‹ standard deviation of triplicates.
the photosynthetic rate of S. quadricauda Berb 614, B
Di€erence between means of various concentrations of test
while the presence of 0.2 mg/l of glyphosate did not in- herbicide on algal growth by one-way ANOVA followed by
hibit algal photosynthetic rate (Table 4). The presence of DuncanÕs mutliple range test at P < 0.05. Control and treat-
2 mg/l of 2,4-D reduced the algal photosynthetic rate to ments, statistically identical, are marked by the same letter.
C
about 80% of the control, while the presence of 0.2 mg/l ND: non-detectable.
180 P.K. Wong / Chemosphere 41 (2000) 177±182

Table 5
Chlorophyll-a content of Scenedesmus quadricauda Berb 614 in the presence of various concentrations of 2,4-D, glyphosate and
paraquat
Herbicide Concentration (mg/l) Chlorophyll-a contentA (10ÿ9 mg/cell)
Log phase Stationary phase
aB
Control ) 2.4 ‹ 0.2 2.8 ‹ 0.4aB
2,4-D 0.02 3.0 ‹ 0.3b 3.2 ‹ 0.5b
0.2 2.8 ‹ 0.2b 3.0 ‹ 0.2b
2 2.4 ‹ 0.3a 2.8 ‹ 0.3a
20 1.0 ‹ 0.1c 1.4 ‹ 0.2c
200 0.2 ‹ 0.1d 0.2 ‹ 0.1d
Glyphosate 0.02 2.5 ‹ 0.3a 3.1 ‹ 0.5b
0.2 2.4 ‹ 0.1a 2.8 ‹ 0.3a
2 1.6 ‹ 0.4e 1.7 ‹ 0.4e
20 0.2 ‹ 0.1d 0.2 ‹ 0.1d
200 0.2 ‹ 0.1d 0.2 ‹ 0.1d
Paraquat 0.02 1.8 ‹ 0.2e 1.8 ‹ 0.1e
0.2 0.8 ‹ 0.2c 0.8 ‹ 0.1f
2 0.2 ‹ 0.1d 0.2 ‹ 0.1d
20 0.2 ‹ 0.1d 0.2 ‹ 0.1d
200 0.2 ‹ 0.1d 0.2 ‹ 0.1d
A
Mean value ‹ standard deviation of triplicates.
B
Di€erence between means of various concentrations of test herbicide on algal growth by one-way ANOVA followed by DuncanÕs
mutliple range test at P < 0.05. Control and treatments, statistically identical, are marked by the same letter.

of paraquat, 2 mg/l of glyphosate or 20 mg/l of 2,4-D through the surface runo€ (Kloeppel et al., 1997). These
reduced the algal photosynthetic rate to about 60% of herbicides e€ectively control their targeted-weed, how-
the control (Table 4). The algal photosynthetic rates ever, it is of great concern to understand their e€ects on
were non-detectable in the presence of 2 mg/l or more of the non-target organisms in aquatic environment. In the
paraquat, 20 mg/l or more of glyphosate and 200 mg/l of present study, S. quadricauda Berb 614, which is com-
2,4-D (Table 4). mon in freshwater ecosystem in Hong Kong, was used to
At stationary phase algal culture treated with low evaluate the toxicity of these herbicides. These herbi-
concentrations (0.02 or 0.2 mg/l) of 2,4-D or (0.02 mg/l) cides have di€erent chemical properties and modes of
of glyphosate showed increases of their chlorophyll-a action (Table 1) on living organisms. Their e€ects on
contents over that of control, while the presence of growth, photosynthesis and chlorophyll-a synthesis of
0.2 mg/l of glyphosate or 2 mg/l of 2,4-D, the algal cul- S. quadricauda Berb 614 were determined in order to
ture produced a similar amount of chlorophyll-a content elucidate the mode of action of these herbicides on the
as the control (Table 5). The presence of 0.02 or 0.2 mg/l algal cells.
of paraquat, 2 mg/l of glyphosate or 20 mg/l of 2,4-D The growth of S. quadricauda Berb 614 was more
signi®cantly reduced the chlorophyll-a contents of the susceptible to the presence of paraquat and less inhibi-
treated algal cultures (Table 5). The presence of 2 mg/l or tion of algal growth was found in the presence of 2,4-D
more of paraquat, 20 mg/l or more of glyphosate, or and glyphosate. Although inhibition of algal growth
200 mg/l of 2,4-D did not show any increase in chloro- can be observed even in the presence of 2 mg/l of three
phyll-a contents from long phase to stationary phase of herbicides, the complete inhibition of algal growth,
growth which indicated the complete inhibition of chlo- except paraquat, required very high concentration
rophyll-a synthesis in the presence of these concentra- (20 mg/l or more) of herbicides. The presence of low
tions of corresponding herbicides (Table 5). concentrations of 2,4-D (0.02 or 0.2 mg/l) and glypho-
sate (0.02 mg/l) stimulated algal growth. The stimulation
may result from a plant growth regulator characteristics
4. Discussion of 2,4-D at low concentration. 2,4-D has been reported
to stimulate growth of plant cells (Draber, 1983).
Herbicides such as 2,4-D, glyphosate and paraquat Glyphosate stimulation may have resulted from its use
which are commonly used in agricultural activities due as carbon or nitrogen source for algal growth (Malik
to their e€ectiveness and low cost. All three herbicides et al., 1989; Marsalek and Rojickova, 1996). Base on the
are water-soluble and their residues in agricultural ®eld results obtained in the present study, the presence of
could be easily translocated into aquatic environments expected environmental concentration (EEC) of the
 P.K. Wong / Chemosphere 41 (2000) 177±182 181

three herbicides (2,4-D about 3 mg/l, glyphosate about Fargasova, A., 1994a. Comparative study on the toxicity and
3 mg/l and paraquat about 0.8 mg/l) (Peterson et al., inhibitory e€ects of inorganic tin compounds on various
1994), glyphosate will slightly inhibit the growth, pho- biological subjects. Biologia 49, 307±311.
tosynthetic rate and chlorophyll-a synthesis of S. quad- Fargasova, A., 1994b. Comparative study of plant growth
hormone (herbicide) toxicity in various biological subjects.
ricauda Berb 614 and paraquat will signi®cantly inhibit
Ecotoxicol. Environ. Safety 29, 359±364.
the three tested parameters of the alga, whereas no in- Fargasova, A., 1994c. Toxicity determination of plant growth
hibitory e€ects of 2,4-D to the alga will be observed. hormones on aquatic alga ± Scenedesmus quadricauda. Bull.
In conclusion, among the three tested herbicides Environ. Contam. Toxicol. 52, 706±711.
paraquat was more toxic than 2,4-D and glyphosate to Fargasova, A., Kizlink, J., 1996. Organotin compounds on the
the growth, photosynthesis and chlorophyll-a synthesis growth of the freshwater alga Scenedesmus quadricauda.
of S. quadricauda Berb 614. The signi®cant inhibitory Ecotoxicol. Environ. Safety 34, 156±159.
e€ects on the algal growth, photosynthesis and chloro- Hosetti, B.B., Shivaraj, K.M., Patil, H.S., 1993. Toxicity of
phyll-a synthesis were only observed in the presence of cobalt chloride on Scenedesmus quadricauda (Turp) Breb
EEC of glyphosate and paraquat (Peterson et al., 1994), during sewage puri®cation. Indian J. Exp. Biol. 31, 627±731.
Ibrahim, E.A., 1990. The in¯uence of the herbicide paraquat
whereas the presence of EEC of 2,4-D (Peterson et al.,
``gramoxone'' on growth and metabolic activity of three
1994) was not inhibitory to the alga tested in the present chlorophytes. Wat. Air Soil Pollut. 51, 89±104.
study. The application of glyphosate and paraquat in Irmer, U., Wachholz, I., Schafer, H., Lorch, D.W., 1986.
agricultural activities should be critically reviewed. In In¯uence of lead on Chlamydomonas reinhardii Danegard
addition, the presence of low concentrations (i.e., 0.02± (Volvocales, Chlorophyta): accumulation, toxicity and ul-
0.2 mg/l) of 2,4-D, glyphosate and paraquat could be trastructural changes. Environ. Exp. Bot. 26, 263±267.
detected by S. quadricauda Berb 614 showing either Jensen, A., 1978. Chlorophyll and carotenoids. In: Hellebust,
stimulating or inhibiting e€ects (Tables 3±5), thus the J.A., Craigie, J.S. Handbook of Phycological Methods:
alga is a good bio-indicator of herbicide contamination Physiological and Biochemical Methods. Cambridge Uni-
in aquatic environments in Hong Kong. However, in versity Press, London, pp. 59±70.
Karim, M.A. (Ed.), 1997. Pesticide Pro®les: Toxicity, Environ-
order to obtain more precise and conclusive toxicologi-
mental Impacts, and Fate. CRC Lewis Publishers, Boca
cal data on the applications of these herbicides, similar Raton.
study using other freshwater algae or plants should be Kasai, F., Hatakeyama, S., 1993. Herbicide susceptibility in
conducted. two green algae, Chlorella vulgaris and Selenastrum capri-
cornutum. Chemosphere 27, 899±904.
Kloeppel, H., Koerdel, W., Stein, B., 1997. Herbicide transport
Acknowledgements by surface runo€ and herbicide retention in a ®lter strip-
rainfall and runo€ simulation studies. Chemosphere 35,
The authors wish to thank Miss T.T. Chong for her 129±141.
Kobraei, M.E., White, D.S., 1996. E€ects of 2,4-Dichlorophen-
preliminary work on the present study. This work was
oxyacetic acid on Kentuckey algae. Arch. Environ. Contam.
partially supported by the Student Campus Working Toxicol. 31, 571±580.
Scheme of United College and Direct Research Grant of Laurent, D.St., Blaise, C., Macquarrie, P., Scroggins, R.,
The Chinese University of Hong Kong. Trottier, B., 1992. Comparative assessment of herbicide
phytotoxicity to Selenastrum capricornutum using micro-
plate and ¯ask bioassay procedures. Environ. Toxicol. Wat.
References Qual. 7, 35±48.
Malik, J., Barry, G., Kishore, G., 1989. The herbicide glypho-
Brown, L.S., Lean, D.R.S., 1995. Toxicity of selected pesticides sate. Biofactor 2, 17±25.
to lake phytoplankton measured using photosynthetic Marsalek, B., Rojickova, R., 1996. Stress factors enhancing
inhibition compared to maximal uptake rates of phosphate production of algal exudates: A potential self-protective
and ammonium. Environ. Toxicol. Chem. 14, 93±98. mechanism? J. Biosci. C 51, 646±650.
Bussieres, D., Cote, R., Richard, C., St-Pierre, E., 1995. Long- Moorhead, D.L., Kosinski, R.J., 1986. E€ects of atrazine on
term copper toxicity on Scenedesmus quadricauda and the productivity of arti®cial stream algal communities. Bull.
complexing polypeptides. Wat. Qual. Res. J. Can. 30, Environ. Contam. Toxicol. 37, 330±336.
265±276. Muromtzev, G.S., Borvko, A.V., Zolnikova, N.V., 1975.
Draber, W., 1983. Plant growth regulators. In: Buchel, K.H. Chlorella as an indicator for detection of microorganisms
Chemistry of Pesticides. Wiley, New York, pp. 393±405. producing phytotoxicity substances. In: Coulson, F., Korte,
Fairchild, J.F., Ruessler, D.S., Haverland, P.S., Carlson, A.R., F. (Eds.), Pesticide. Georg Thieme Publishers, Stuttgart,
1997. Comparative sensitivity of Selenastrun capricornutum pp. 672±674.
and Lemma minor to sixteen herbicides. Arch. Environ. Okay, O.S., Gaines, A., 1996. Toxicity of 2,4-D acid to
Contam. Toxicol. 32, 353±357. phytoplankton. Wat. Res. 30, 688±696.
Fargasova, A., 1993. E€ects of ®ve toxic metals on the alga Peterson, H.G., Boutin, C., Martin, P.A., Freemark, K.E.,
Scenedesmus quadricauda. Biologia 48, 301±304. Ruecker, N.J., Moody, M.J., 1994. Aquatic phyto-toxicity
182 P.K. Wong / Chemosphere 41 (2000) 177±182

of 23 pesticides applied at expected environmental concen- Mechanisms of Pesticide Action. American Chemical Soci-
trations. Aquatic Toxicol. 28, 275±292. ety, Washington, DC, pp. 156±168.
Reddy, G.N., Prasad, M.N.V., 1994. Cadmium toxicity of Tisler, T., Zagorc-Koncan, J., 1997. Comparative assessment of
Scenedesmus quadricauda: proteins and protein phosphory- toxicity of phenol, fomaldehyde, and industrial wastewater
lation changes. Biochem. Arch. 10, 185±188. to aquatic organisms. Wat. Air Soil Pollut. 97, 315±322.
Saenz, M.E., Accorinti, J., Del-Carment-Tortorellu, M., 1993. Wong, P.K., Chang, L., 1988. The e€ects of 2,4-D herbicide and
Toxicity of paraquat to a green alga Scenedsmus acutus. organophosphorus insecticides on growth, photosynthesis,
J. Environ. Sci. Health B 28, 193±204. and chlorophyll a synthesis of Chlamydomonas reinhardtii
Sullivan, D.S., Sullivan, T.P., Bisalputra, T., 1981. E€ects of (mt+). Environ. Pollut. 55, 179±189.
Roundupâ herbicide on diatom populations in aquatic Wong, P.K., Chang, L., 1991. E€ects of copper, chromium and
environment of a coastal forest. Bull. Environ. Contam. nickel on growth, photosynthesis and chlorophyll a synthe-
Toxicol. 26, 91±96. sis of Chlorella pyrenoidosa 251. Environ. Pollut. 72,
Sumida, S., Ueda, M., 1974. Chlorella as a model system to 127±139.
study herbicidal mode of action and its application to a new Worthing, C.R., Walker, S.B., 1987. The Pesticide Manual: A
herbicide, O-ethyl-O-(3-metyl-6-nitrophenyl)-N-sec-butyl World Compendium. The British Crop Protection Council,
phosphorothioamidate (S-2846). In: Kohn, G.K. (Ed.), Thornton Heath.