Toxicity of the antimicrobial compound triclosan and formation of the metabolite methyl-triclosan in estuarine systems

Triclosan, a commonly used antimicrobial compound, has been measured in aquatic systems worldwide. This study exposed marine species to triclosan to examine effects primarily on survival and to investigate the formation of the degradation product, methyl-triclosan, in the estuarine environment. Acute toxicity was assessed using the bacterium Vibrio fischeri, the phytoplankton species Dunaliella tertiolecta, and three life stages of the grass shrimp Palaemonetes pugio. P. pugio larvae were more sensitive to triclosan than adult shrimp or embryos. Acute aqueous toxicity values (96 h LC50) were 305 microg/L for adult shrimp, 154 microg/L for larvae, and 651 microg/L for embryos. The presence of sediment decreased triclosan toxicity in adult shrimp (24 h LC50s were 620 microg/L with sediment, and 482 microg/L without sediment). The bacterium was more sensitive to triclosan than the grass shrimp, with a 15 min aqueous IC50 value of 53 microg/L and a 15 min spiked sediment IC50 value of 616 microg/kg. The phytoplankton species was the most sensitive species tested, with a 96 h EC50 value of 3.55 microg/L. Adult grass shrimp were found to accumulate methyl-triclosan after a 14-day exposure to 100 microg/L triclosan, indicating formation of this metabolite in a seawater environment and its potential to bioaccumulate in higher organisms. Triclosan was detected in limited surface water sampling of Charleston Harbor, SC at a maximum concentration of 0.001 microg/L, substantially lower than the determined toxicity values. These findings suggest triclosan poses low acute toxicity risk to estuarine organisms; however, the potential for chronic, sublethal, and metabolite effects should be investigated.     

Influence of light in acute toxicity bioassays of imidacloprid and zinc pyrithione to zooplankton crustaceans

The acute toxicity of imidacloprid, a neonicotinoid insecticide, and zinc pyrithione (Zpt), a biocide used in anti-dandruff shampoos and protective antifouling paints, to three species of ostracods and two waterfleas, including Daphnia magna, was determined and compared under light and dark conditions. Under normal laboratory conditions, UV light had no significant influence on the outcome of toxicity bioassays, although in the case of imidacloprid both EC(50) and LC(50) calculated values were twice as high under the light as in the dark. No influence of UV light was observed on bioassays conducted with Zpt, in spite of the fast aqueous photolysis exhibited by this compound. Imidacloprid 48-h LC(50) for cladocerans (65-133mg/L) were two orders of magnitude higher than for ostracods (301-715microg/L); values of EC(50) for cladocerans and ostracods were 2-6mg/L and 3-16microg/L, respectively. Toxicity of Zpt to both ostracod and cladoceran species appears to be similar, with 48-h LC(50) in the range 137-524 and 75-197microg/L for ostracods and cladocerans, respectively, and similar values for EC(50)s. The mortality endpoint (LC(50)), however, is not a reliable predictor of the effects of imidacloprid under field situations (e.g. rice paddies), because the paralysis effect induced by this insecticide takes place at much lower concentrations than those required to cause the death of the animals: regardless of the taxa, differences as large as 100- or 600-fold were observed between the EC(50) and LC(50) for the same exposures. As a consequence, immobilization tests and EC(50) values are recommended for this class of compounds, while caution should be exercised in environmental risk assessments of this and possibly other related neonicotinoid insecticides with similar activity.     

Piscicidal and anti-acetylcholinesterase activity of Euphorbia royleana stem bark extracts against freshwater common predatory fish Channa punctatus

The present paper deals with the piscicidal activity of different solvent extracts of Euphorbia royleana stem bark against freshwater predatory fish Channa punctatus, which is commonly present in shrimp and carp culture ponds. The rank of order of toxicity of different solvent extract was: diethyl ether (LC(50) (96h) 31.76mg DW/L) > chloroform (LC(50) (96h) 56.26mg DW/L) > methanol (LC(50) (96h) 56.80mg DW/L) > acetone (LC(50) (96h) 65.77mg DW/L). There was a significant (P < 0.05) negative correlation between LC values and exposure periods, thus increase in exposure time. The LC(50) values of diethyl ether extract of E. royleana stem bark decrease from 81.78mg DW/L (24h) to 31.76mg DW/L (96h). Similar trend were also observed in case of chloroform, methanol and acetone extracts. Exposure of sub-lethal doses (40% and 80% of LC(50)) of the diethyl ether extract for 24h or 96h caused significant inhibition in the activity of enzyme acetylcholinesterase (AChE) in liver and muscle tissue of fish C. punctatus. There was a significant recovery observed in the activity of enzyme acetylcholinesterase in both the tissues of fish after 7th day of the withdrawal of treatment. Thus, the diethyl ether extract of E. royleana stem bark can be used for control of unwanted predatory fish C. punctatus from fish culture ponds. Their toxicological action is due to their anti-acetylcholinesterase enzyme activity, reversibility in their action is advantageous factor for their use as environmentally safe piscicide in aquatic bodies.     

大型溞生物测试法对农药溴氰菊酯的毒性评价

溴氰菊酯对大型(氵蚤)的半数运动受抑制浓度值(EC_(50)),24、48和96小时分别为0.113、0、31和0.03μg/L;半数致死浓度(LC_(50))值,24、48和96小时分别为0.133、0.038和0.01μg/L,其毒性比汞离子还大。作者认为,大型(氵蚤)生物测试法是快速、灵敏、经济、可靠的,值得推广应用。     

Effects of chlordane and lindane on testosterone and vitellogenin levels in green neon shrimp (Neocaridina denticulata)

The purpose of this study was to investigate the acute toxicity of chlordane and lindane as well as their endocrine disruption effect on green neon shrimp (Neocaridina denticulata), a common habitant in freshwater system of eastern Asia and Hawaii. First, the organisms were exposed to chlordane and lindane to estimate the 96-h LC(50)(96-h median lethal concentration). Then, levels of testosterone and vitellogenin in hemolymph of N. denticulata after exposure to sublethal concentrations of chlordane (1 ng/L and 10 ng/L) and lindane (0.1 microg/L and 1 microg/L) were also examined. The 96-h LC(50) values obtained from the results of acute exposure were 127.03 (130.11-122.35) ng/L and 9.36 (8.00-10.96) microg/L for chlordane and lindane, respectively. Furthermore, reductions of testosterone concentration were observed in both chlordane- and lindane-treated shrimps, whereas induction of vitellogenin-like protein was only apparent in chlordane-treated shrimps. Thus, it is concluded that chlordane and lindane may probably show some disruption endocrine functions on N. denticulata.     

Tissue-specific Cu bioaccumulation patterns and differences in sensitivity to waterborne Cu in three freshwater fish: rainbow trout (Oncorhynchus mykiss), common carp (Cyprinus carpio), and gibel carp (Carassius auratus gibelio)

Rainbow trout (Oncorhynchus mykiss), common carp (Cyprinus carpio) and gibel carp (Carassius auratus gibelio) were exposed to copper (1-20 microM) in softened Antwerp City tap water at pH 7.3 +/- 0.1 and with a water hardness of 292.4 +/- 8.1 mg/L CaCO(3) (Ca 100.8 +/- 3.0 mg/L; Mg 11.0 +/- 0.2 mg/L). LC50s (96 h) were determined and copper accumulation in gills, liver, and kidney assessed over a 10-day period. Rainbow trout (96 h LC50: 3.3 microM/210 microg/L) were three times more sensitive to Cu exposure than common carp (96 h LC50: 10.4 microM/661 microg/L) and almost seven times more sensitive than gibel carp (96 h LC50: 22.0 microM/1398 microg/L). After 96 h, the incipient lethal level (ILL) was reached for common carp, and by the end of the experiment (>120 h) also for rainbow trout. The ILL was never reached for gibel carp. Survival analysis confirmed the differences in sensitivity shown by the 96 h LC50 values. At 1 microM Cu, the relative risk to die was six to seven times greater for rainbow trout as for common or gibel carp, respectively, while it was 9000 and 19,000 times greater at 5 microM Cu. Only the environmental Cu concentrations contributed significantly (P < 0.001) to the Time-To-Death (TTD). Tissue Cu concentrations did not relate to TTD. Among species, a clear difference in metal handling was apparent, with high liver residues and liver accumulation rates for the most sensitive species, the rainbow trout, and lower liver but higher kidney residues and kidney accumulation rates for the most resistant species, the gibel carp. Gill concentrations and accumulation rates were lowest in the sensitive rainbow trout.     

Acute toxicity, behavioral changes, and histopathological effects of deltamethrin on tissues (gills, liver, brain, spleen, kidney, muscle, skin) of Nile tilapia (Oreochromis niloticus L.) fingerlings

Deltamethrin, a synthetic pyrethroid contaminating aquatic ecosystems as a potential toxic pollutant, was investigated in the present study for acute toxicity. The purpose of this study was to evaluate LC(50) values of deltamethrin on Nile tilapia (Oreochromis niloticus L.) fingerlings and investigate histopathological responses of fish exposed to deltamethrin. The 48 h LC(50) value for Nile tilapia fingerlings was estimated as 4.85 microg/L using static test system. In addition, behavioral changes at each deltamethrin concentration were observed closely. All fish, exposed to 5 microg/L deltamethrin revealed severe morphological alterations in the gills and liver. In the gills hyperemia, fusion of secondary lamellae and telangiectasis were observed; whereas hydropic degenerations in liver were observed in all examined fish. The results are significant for reporting acute deltamethrin toxicity in terms of behavioral and histopathological changes: Deltamethrin is highly toxic to fingerlings.     

Piperonyl butoxide enhances the bioconcentration and photoinduced toxicity of fluoranthene and benzo[a]pyrene to larvae of the grass shrimp (Palaemonetes pugio)

Piperonyl butoxide (PBO) is a commonly used synergist in many pyrethroid formulations due to its ability to interfere with cytochrome P450 (CYP) monooxygenases. Because PBO can co-occur in the estuarine environment with polycyclic aromatic hydrocarbons (PAHs), a class of compounds metabolized by CYP isozymes, the overall objective of this study was to investigate the influence of PBO on the bioconcentration and photoinduced toxicity of two common PAH contaminants, fluoranthene (FLU) and benzo[a]pyrene (BaP), on the larvae of the grass shrimp (Palaemonetes pugio). PBO alone was not particularly toxic to grass shrimp larvae. In dark exposures and under simulated sunlight (UV-A=211.0+/-7.0 microW/cm(2), UV-B=9.8+/-2.4microW/cm(2)), 96-h LC(50) values were similar (814.4 and 888.6 microg/L, respectively), suggesting that PBO toxicity is not enhanced in the presence of sunlight. The presence of sublethal concentrations of PBO in single PAH toxicity tests increased the bioconcentration of the two tested PAHs, and these increases were greatest at the lowest tested PAH concentrations. Mean bioconcentration factors (BCF) at the three lowest FLU and BaP treatments increased 14.3- and 7.1-fold, respectively, in the low PBO (127 microg/L) exposure compared to that of the no PBO exposure. Under simulated sunlight, PBO exposure also increased the photoinduced toxicity of the two tested PAHs, and this increase occurred in a PBO concentration-dependent fashion. For FLU, 96-h LC(50) values decreased from 2.35 microg/L in the absence of PBO to 0.76 microg/L in the high PBO (256 microg/L) exposure. For BaP, 96-h LC(50) values similarly decreased from 1.02 microg/L in the absence of PBO to 0.30microg/L in the high PBO exposure. The presence of PBO also influenced the PAH tissue residue-response relationship, but in different ways for FLU and BaP. For FLU, slopes of the tissue residue-response relationship decreased in the presence of PBO, and for BaP, there was a trend towards increased slopes in the presence of PBO. These results demonstrate that sublethal levels of PBO increase the bioconcentration and photoinduced toxicity of certain PAH in grass shrimp larvae, and underscore the need to consider the potential for PBO to synergize the toxicity of co-occurring environmental contaminants in future risk assessments.     

Effects of cyanobacterial biomass and purified microcystins on malformations in Xenopus laevis: teratogenesis assay (FETAX)

Xenopus laevis (African clawed frog) embryos in a 96-h teratogenesis assay (FETAX) were exposed to 0-250 microg/L and 500 microg/L of purified microcystin-LR (MCYST-LR) for the estimation of lethality, as well as to equivalent concentrations of biomass containing MCYST-LR (natural water bloom dominated by Microcystis aeruginosa) and biomass without MCYST-LR (bloom dominated by Microcystis wesenbergii). The highest tested concentrations of purified MCYST-LR caused up to 30% lethality after a 96-h exposure, corresponding to a LC(25) of 380 microg/L. Cyanobacterial biomass containing MCYST-LR caused significant lethality up to 50% at the highest tested concentrations (300 mg/L, i.e., 250 microg/L of MCYST-LR). The estimated 96-h LC(25) values varied from 125 mg/L (biomass containing MCYST-LR) up to 232 mg/L (biomass without MCYST-LR). A statistically significant increase in the number of malformed embryos was observed after exposure to cyanobacterial samples. Purified MCYST-LR at and above 25 microg/L significantly increased the number of malformations, with 53% of surviving embryos malformed in the highest tested concentration, 250 microg/L (EC(25) = 27 microg/L). Exposure to the highest concentration of MCYST-LR containing biomass resulted in more than 60% of the embryos being malformed and an EC(25) of 52 mg/L (i.e., 43 microg of MCYST-LR/L). Cyanobacterial biomass with no natural microcystin also induced substantial malformations-about 50% aberrant embryos at the highest concentration, 300 mg/L (EC(25) = 75 mg/L). External additions of purified MCYST-LR to the biomass that was originally without microcystins resulted in a slight additional increase in the rate of malformations (80% at the highest concentration, 300 mg of biomass plus 250 microg of MCYST-LR per liter). A comparison of lethality and effects on malformations (teratogenic index, TI = LC(25)/EC(25)) showed that all samples had significant teratogenic potential in the FETAX assay (TI(MCYST-LR) = 14; TI for biomass with and without microcystin ranged between 2.4 and 3.1, respectively). We conclude that cyanobacterial water blooms can significantly alter the normal development of amphibian embryos.     

The effects of metals on embryo-larval and adult life stages of the sea urchin, Diadema antillarum

Since the massive population decline of the long-spined sea urchin, Diadema antillarum, in the early 1980s, the dynamics of coral reef ecosystems in the Caribbean have changed tremendously. The absence of D. antillarum, once a keystone herbivore, has led to macroalgal dominance in many of these reef communities. D. antillarum is not only important ecologically, but may also be a sensitive bioindicator species for toxicant exposure. Echinoderm larval development tests were conducted with D. antillarum exposed to elevated levels of aqueous copper (Cu), silver (Ag), nickel (Ni), or selenium (Se). All metals significantly affected larval development, based on normal development to the pluteus stage. The EC50s based on dissolved metal concentrations were 11 microg/L Cu, 6 microg/L Ag, 15 microg/L Ni, and 26 microg/L Se. Adult sea urchins were exposed to aqueous copper under flow through conditions for 96 h. The 96-h LC50 for this exposure was 25 microg/L dissolved Cu. Additionally, behavioral and physiological disturbance was observed. The physiological responses included both acid-base balance disturbance, as evidenced by reduced coelomic fluid pH and apparent ionoregulatory effects. In addition, behavioral effects included spatial orientation within the exposure tank, spine closure, and loss of spines. The high sensitivity of both adult and larval D. antillarum to these metals supports the use of this organism as an important biological indicator for metal exposure in marine environments.     

Comparative toxicity of chlorothalonil and chlorpyrifos: Ceriodaphnia dubia and Pimephales promelas

This study was done to characterize responses of Ceriodaphnia dubia Richard and Pimephales promelas Rafinesque exposed to aqueous solutions of chlorothalonil (tetrachloroisophthalonitrile) and chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate). Chlorothalonil (a fungicide) and chlorpyrifos (an insecticide) are intensely used in agricultural, silvicultural, and urban settings. These pesticides may enter aquatic systems through several pathways including rainfall runoff. C. dubia and P. promelas have been used to monitor surface waters and discern the effects of pesticides that contaminate those waters. Modified static renewal exposures (7 or 10 days) with individual solutions of chlorothalonil and chlorpyrifos were used to obtain mortality data for C. dubia and P. promelas, from which potency curves were derived, as well as sublethal effects data (reproduction or growth). In these experiments P. promelas was more sensitive to chlorothalonil, and C. dubia was more sensitive to chlorpyrifos. Lower and upper thresholds (i.e., LC(0) and LC(100)) for 7-day P. promelas exposures to chlorothalonil were 14.4 and 30.8 microg/L, respectively, in contrast to the lower and upper threshold values, 103 and 210 microg/L, respectively, for C. dubia. Ten-day exposures of C. dubia to chlorpyrifos resulted in lower and upper threshold values of 0.05 and 0.09 microg/L, whereas 10-day exposures of P. promelas to chlorpyrifos yielded threshold values of 26 and 274 microg/L. The results of this study illustrated differences in species' sensitivities to chlorothalonil and chlorpyrifos as well as differences in the duration of the exposure necessary to illustrate effects that might be elicited from pesticide exposures.     

Chronic effects of p-nonylphenol on survival and reproduction of Daphnia galeata: multigenerational life table experiment

The chronic effect of p-nonylphenol on survival and reproduction for two generations of the freshwater cladoceran Daphnia galeata was examined by life table experiments. The effects on survival and reproduction were used as the intrinsic rate of natural increase, r, with the Euler-Lotka equation and were analyzed with a simple mathematical model (a power function). The population-level EC(50), the concentration of a substance that reduces the intrinsic rate of natural increase by 50%, was estimated as 65.2 microg/L for the first generation and 81.5 microg/L for the second generation. No transgenerational effect that reinforces adverse responses in the offspring generation has been detected. From a 48-h immobility test an acute LC(50) was estimated to be 60.8 microg/L. The acute LC(50) is a good indicator of the chronic population-level effects of this chemical to this species.     

Lethal and sublethal responses of an aquatic insect Culex quinquefasciatus (Diptera: Culicidae) challenged with individual and joint exposure to dissolved sodium selenate and methylmercury chloride

Pollutants rarely occur alone in the natural environment, and few studies have focused on the potential interactions between metals or metalloids. In this study an aquatic insect, the southern house mosquito (Culex quinquefasciatus: Diptera), was used to test the individual and joint effects of dissolved sodium selenate (Se) and methyl mercury chloride (MeHg). We conducted ovipositional preference tests and 14-day chronic toxicity studies to determine lethal and sublethal responses of C. quinquefasciatus to a range of Se and MeHg concentrations and mixtures. No evidence was found for female ovipositional preference in field trials using artificial ponds. Larvae were more sensitive to MeHg than Se, with LC(50) values of 30 microg/L (CI = 28-31 microg/L) and 11 mg/L (CI = 10-12 mg/L) respectively. In addition, larval survival was significantly reduced at concentrations as low as 25 microg/L of MeHg and 8 mg/L of Se. A synergistic interaction was observed in the toxicity of the Se-MeHg mixtures to C. quinquefasciatus larvae. Larval mosquito survival was significantly reduced at 7.5 microg/L MeHg + 2.75 mg/L Se and an LC(50) value of 9 microg/L MeHg + 3.4 mg/L Se was determined for a fixed ratio mixture. The rate of growth of the larvae was analyzed using a Growth Index that provided a sensitive measure of the developmental effects of toxicant exposure. Sodium selenate at concentrations as low as 2 mg/L caused a significant decrease in growth between larvae in treatment versus control solutions after only 4 days. Similarly, MeHg at concentrations as low as 25 microg/L and a Se-MeHg mixture of 3 microg/L MeHg plus 1.1 mg/L Se caused significant growth reductions after only 2 and 3 days, respectively. These are the first reported survival and developmental data for an aquatic insect exposed to MeHg and Se-MeHg mixtures.     

Changes in nitrogen metabolism of the freshwater fish Cirrhinus mrigala following exposure to cypermethrin

We studied the effect of a synthetic pyrethroid insecticide, cypermethrin, on some aspects of nitrogen metabolism in the gills, liver, and muscle tissue of the Indian major carp, Cirrhinus mrigala. Free amino acid levels and protease activity showed an elevation in all tissues following exposure to lethal (5.13 microg/L) and sublethal (1.026 microg/L) doses of cypermethrin, with significant changes noticed on day 4. The normal condition was restored only at a sublethal concentration.     

Histological alterations in the ultimobranchial gland of teleost Heteropneustes fossilis in response to chlorpyrifos treatment

In this study, an experiment was performed on Heteropneustes fossilis for short-term (1.76 mg/L chlorpyrifos, i.e., 0.8 of 96-h LC50) and long-term (0.44 mg/L chlorpyrifos, i.e., 0.2 of 96-h LC50) exposure. The fish were sacrificed after 24, 48, 72 and 96 h in the short-term experiment and after 7, 14, 21 and 28 days in the long-term experiment. On these intervals, blood was collected and analysis of serum calcium was done. Ultimobranchial glands were also fixed for histological study. The serum calcium levels of H. fossilis exhibit a decline after 24 h following exposure to chlorpyrifos. This decrease continues until the end of the experiment (96 h). The serum calcium levels of chronically exposed fish exhibit a decrease on day 7. Thereafter, the levels continue to fall progressively until the end of the experiment (28 days). The ultimobranchial gland of chlorpyrifos treated fish exhibits no histological change up to 48 h. After 72 h, there is a decrease in the staining response of cytoplasm of the ultimobranchial cells. The nuclear volume of these cells is slightly decreased. After 96 h following chlorpyrifos exposure, these changes become exaggerated. In chlorpyrifos-treated fish there is no change in the histological structure of the ultimobranchial gland up to 14 days. After 21 days, the cytoplasm of ultimobranchial cells stain feebly and the nuclear volume of these cells exhibits a decrease. Following 28 days treatment, the nuclear volume of these cells records a further decrease and the gland depicts vacuolization and degeneration at certain areas.     

Comparison of the acute toxicity for gamma-cyhalothrin and lambda-cyhalothrin to zebra fish and shrimp

Gamma-cyhalothrin 15CS (GCH) contains only the active stereoisomer of the two isomers found in lambda-cyhalothrin 25EW (LCH). GCH (0.5 x rate) provides equivalent overall insect control as LCH (1 x rate). Both formulations showed high acute toxicity to zebra fish (Brachydanio rerio H.B.) and shrimp (Macrobrachium nippoensis de Haan). The 96-h LC(50(zebra fish,GCH)) is 1.93 microg a.i/L and LC(50(zebra fish,LCH)) is 1.94 microg a.i/L. LC(50(shrimp,GCH)) is 0.28 microg a.i./L and LC(50(shrimp,LCH)) 0.04 microg a.i./L. This indicates that the toxicity to shrimp is likely stereochemistry-dependent. The fates of GCH and LCH are similar in laboratory simulated rice paddy water and their concentrations decrease rapidly, with no GCH or LCH detected after 3 or 4 days. Both are toxic to shrimp in a simulated paddy irrigation reservoir even though treated return water is diluted 5 times. No shrimp fatality is shown in the GCH-treated paddy water after a 4-day holding period, and longer than 5 days is necessary to reach a zero fatality rate for LCH. This is compatible with the 7-day water holding period considered reasonable in agricultural practice.     

Effects of the pyrethroid insecticide,cypermethrin, on a freshwater community studied under field conditions. I. Direct and indirect effects on abundance measures of organisms at different trophic levels

The effects of the pyrethroid insecticide cypermethrin on a natural freshwater community were studied in small in situ enclosures over an 11-day period. The experiment was conducted in a eutrophic lake using a regression design that included three untreated controls and a gradient of six unreplicated cypermethrin concentrations, ranging from 0.01 to 6.1 microg/l. This paper is the first in a series of two, and describes the fate of cypermethrin and its effects on the abundance of crustaceans, rotifers, protozoans (cilliates and heterotrophic nanoflagellates (HNF)) and bacteria and the biomass of periphytic and planktonic algae. The concentration of cypermethrin decreased quickly during the experiment, with a half-life of 48 h for the total and 25 h for the dissolved fractions of cypermethrin, respectively. Cypermethrin proved to be acutely toxic to crustaceans in enclosures receiving nominal cypermethrin concentrations of >/=0.13 microg/l. No Effect Concentration (NEC) and median Effect Concentration (EC(50)) for the total crustacean community and cladoceran and copepod subgroups ranged between 0.02-0.07 and 0.04-0.17 microg/l, respectively, with copepods being less sensitive than cladocerans. The abundance of rotifers, protozoans and bacteria and the chlorophyll-a concentration of planktonic and periphytic algae was significantly related to the concentration of cypermethrin. All groups proliferated within 2-7 days after the cypermethrin application in those enclosures where the abundance of crustaceans was seriously affected by cypermethrin (i.e. >/=0.13 microg/l). We hypothesise that the proliferation of rotifers, protozoans, bacteria and algae was due to a reduced grazer control from crustaceans and thereby mediated indirectly by cypermethrin. The results of this experiment provide knowledge on how an entire microplankton community may respond to pyrethroids in nature, and the indirect effects observed on the community clearly demonstrates the necessity of multispecies field experiments in ecotoxicological risk assessment.     

In vitro screening for antitumour activity of Clinopodium vulgare L. (Lamiaceae) extracts

Aqueous extract of Clinopodium vulgare L. showed strong antitumour activity when tested in vitro on A2058 (human metastatic melanoma), HEp-2 (epidermoid carcinoma, larynx, human) and L5178Y (mouse lymphoma) cell lines-6 h after treatment disintegration of the nuclei and cell lysis started. Applied at a concentration of 80 microg/ml it reduced the cell survival to 1.0, 5.6 and 6.6%, respectively. The concentrations of aqueous extract inhibiting the growth of A2058, HEp-2 and L5178Y cells by 50% (IC50 values) were calculated to be 20, 10 and 17.8 microg/ml respectively. Two groups of active substances were detected: the first one, probably combining glycosides, influenced adhesion, while the second one caused massive cell vacuolisation. The chloroform extract, which contained ursolic acid and gentriacontan had also cytotoxic, however a little bit weaker effect. All changes observed were irreversible.     

Isolation of praeruptorins A and B from Peucedanum praeruptorum Dunn. and their general pharmacological evaluation in comparison with extracts of the drug

The root of Peucedanum praeruptorum Dunn. was extracted with solvents at different polarity obtaining three chemical fractions: aqueous (H2O), n-butanol (BuOH) and ethyl acetate (AcOEt). From AcOEt praeruptorins A and B were isolated by column chromatography on silica gel, using toluene/ethyl acetate as eluent, and identified by 1H and 13C NMR analysis. The extracts and the praeruptorins were tested for gross behavioural effects and acute toxicity in mice; the cytotoxicity on Artemia salina Leach and the antimicrobial activity were also evaluated. None of the tested substances evoked behavioural effects or acute toxicity after oral administration in mice; delayed mortality was observed with AcOEt and praeruptorin A only after intraperitoneal administration of high doses (1 g/kg). In Artemia salina test AcOEt, and praeruptorins A and B had LC50 values of 40.2, 121.2 and 34.5 microg/ml, respectively. AcOEt and praeruptorin A showed antimicrobial activity on Streptococcus agalactiae; their MIC values were 250 and 100 microg/ml, respectively.     

Toxicity of cylindrospermopsin to the brine shrimp Artemia salina: comparisons with protein synthesis inhibitors and microcystins.

The Artemia salina bioassay was successfully applied to the analysis of the hepatotoxic cyanobacterial alkaloid and protein synthesis inhibitor, cylindrospermopsin. A dose-dependent response in mortality was observed for purified cylindrospermopsin and LC(50) values decreased with time from 8.1 to 0.71 microg/ml(-1), between 24 and 72 h, respectively. Cylindrospermopsin was slightly less potent than micro cystin-LR, with similar LC(50) values on a gravimetric basis, but was more toxic to A.salina than the protein synthesis inhibitors, cycloheximide, chloramphenicol and tetracycline. Cylindrospermopsin-containing strains of the cyanobacterium Cylindrospermopsis raciborskii were found to be toxic to A.salina and the LC(50) concentration for these strains over time was greater than the LC(50) for purified cylindrospermopsin, with the exception of C. raciborskii strain CR1.