E. Andreu-Moliner

Eel ATPase activity as biomarker of thiobencarb exposure

European eels (Anguilla anguilla) were exposed to a sublethal thiobencarb concentration of 0.22 mg/L in a flow-through system for 96 h. Mg(2+) and Na(+)-K(+) adenosine triphosphatase (ATPase) activities were evaluated in gill and muscle tissues at 2, 12, 24, 48, 72, and 96 h of thiobencarb exposure. Gill ATPase activities were rapidly inhibited from 2h of contact onward. Highest inhibition was registered for Na(+), K(+)-ATPase (85%) from 2 to 12h. Both Mg(2+) and total ATPase were inhibited (>73%) during the first hours of toxicant exposure. At the end of the exposure period (96 h) ATPase activities were still different from those of the controls (>50%). Significant inhibition was detected in Na(+), K(+)-ATPase activity (80%) in muscle tissue after 2h and it was maintained over the entire exposure time. However, Mg(2+)-ATPase and total ATPase showed only perturbations after 2 h of exposure. Eels were exposed to 0.22 mg/L of thiobencarb for 96 h and then a recovery period in herbicide-free water was allowed for 192 h. Gill and muscle samples were removed at 8, 24, 72, 96, 144, and 192 h and ATPase activity was evaluated. Following 144 h of recovery, Mg(2+)- and Na(+), K(+)-ATPase activities, as well as total ATPase activity, in gills of those animals previously exposed to 0.22 mg/L of thiobencarb were still significantly different compared to controls. Thiobencarb seems to act to alter the ionic profiles. Since ion-dependent ATPases are known to regulate the influx and efflux of ions across the membrane to maintain the physiological requirements of the cells, the inhibition of Na(+), K(+)-ATPase probably induced osmoregulatory perturbations. On the other hand, thiobencarb exposure causes increases in the muscle water content of A. anguilla. The results indicated that water content increased significantly (>100% higher than the controls) during the first 24 h of exposure.

Thiobencarb toxicity and plasma AChE inhibition in the European eel

The acute toxicity of the herbicide thiobencarb (S-4-chlorobenzyl diethylthiocarbamate) was determined for the European eel (Anguilla anguilla). The 24, 48, 72 and 96 hours median lethal concentrations (LC50) were 25.7, 21.7, 17.0 and 13.2 mg/L, respectively. Fish were also exposed to a sublethal thiobencarb concentration (1/60 LC50-96 hr = 0.22 mg/L) during 96 hours in a flow-through system and then an elimination period of 192 hours in clean water was allowed. Eels were removed and blood samples taken out at each exposure time and recovery period in order to evaluate AChE activity. Thiobencarb induced significant inhibitory effects on plasma AChE activity of A. anguilla from the first contact time with the toxicant. This inhibition (under 50% activity) was maintained during the entire exposure period (96 hours) and even those animals transferred to clean water showed plasma AChE activity different from the controls. Differences between total and specific AChE activity were detected during the exposure period. Total AChE activity in the plasma from animals transferred to a medium free of toxicant recovered its normal value while specific AChE activity remained depressed (< 50%) until five days later.

Effect of the insecticide methylparathion on filtration and ingestion rates of Brachionus calyciflorus and Daphnia magna

The freshwater rotifer Brachionus calyciflorus and the cladocera Daphnia magna were exposed to sublethal levels of methylparathion to determine the effect on filtration and ingestion rates. The experiments were performed using the unicellular algae, Nannochloris oculata in a density of 5 × 105 cell/ml. Prior to feeding experiments, preliminary acute toxicity tests were carried out to determine 24-h LC50 values for both species, these values indicated that Daphnia magna was more sensitive to methylparathion acute exposure than Brachionus calyciflorus was. Rates of filtration and ingestion declined with increasing methylparathion concentrations after an exposure of 5 h to this toxicant. The effective concentration at which filtration and ingestion rates were reduced to 50% of those in controls (EC50) were calculed for both species.

Biochemical stress response in tetradifon exposed Daphnia magna and its relationship to individual growth and reproduction.

Environmental risk assessment of chemicals toxicity requires the use of costly and labor-intensive chronic data and short-term tests provide additional information. Energy budget is used by the animals for their growth, reproduction, and metabolism and it is reduced in case of toxic stress. Tetradifon acaricide is frequently used in the European Mediterranean region and it is implicated in aquatic environmental pollution. Previous studies showed that the EC(50)-24 h of tetradifon on Daphnia magna was 8.92 mg/L. Based on that, D. magna were exposed to sublethal tetradifon concentrations of 0.10, 0.18, 0.22 and 0.44 mg/L for five days in order to investigate their effect on intermediate metabolism. Caloric content was determined as biomarker of tetradifon toxicity. Results were analyzed using one-way analysis of variance (ANOVA) and Duncans significant difference test was used to find differences between groups (alpha was set at p=0.05). Daphnids energy content decreased as tetradifon concentration increased. At 120-h caloric content was depleted >51% at pesticide concentrations of 0.18 mg/L and higher. In order to determine a possible link between the 5-d test and the 21-d chronic test, animals under short-term test were exposed to the same pesticide concentrations known to cause adverse effects on reproduction, growth and survival. Results from the present study indicated a good correlation between the proposed 5-day test and daphnid energy budget. Comparison between both, caloric content results and the chronic effect values obtained using life-table studies, suggested a good fit between them. These studies can be used as earlier, predictive and useful tests with comparable results to the classic chronic ones. Our results indicate that caution must be done about the use of tetradifon in the aquatic environment.

Physiological effects of tricyclazole on zebrafish (Danio rerio) and post-exposure recovery.

Short-term effects of tricyclazole on male zebrafish (Danio rerio) physiology were examined joint to the degree of recovery after exposure. Fish were exposed to 142 microg/L (1/100 LC(50)-96 h) of tricyclazole for 7 (Exp.1) and 14 days (Exp.2) and then allowed to recover for 7 or 14 more days, respectively. Whole-body triglycerides, cholesterol, glucose, lactate and total proteins were measured as well as the aspartate aminotransferase (AAT), alanine aminotransferase (AlAT), alkaline phosphatase (AP) and lactate dehydrogenase (LDH) activities as biomarkers of intermediary metabolism; gamma-glutamyl transpeptidase (gammaGT) as biomarker of oxidative detoxification processes and vitellogenin (Vtg) concentration as endpoint for endocrine disruptor effect were also determined. Corpulence factor (k) was calculated. Fungicide exposure in zebrafish resulted in an increased of triglycerides, cholesterol, glucose and lactate levels, however the total protein content did not change. LDH, AlAT and AAT enhanced while AP activity decreased. Corpulence factor (k) also decreased. At the end of the recovery periods cholesterol and glucose levels recovered whereas triglycerides and lactate continued to elevate. Induced disorders on the selected enzymes remained and did not recover at the end of experiments. Fish exhibited significant increases of Vtg during the overall experimental times as a consequence of the fungicide exposure. These findings are of importance in the assessment of the potential risk of new fungicides as tricyclazole on aquatic ecosystems.

Evaluation of a Daphnia magna renewal life‐cycle test method with diazinon

Abstract Acute and chronic toxicity tests with diazinon (diethyl 2‐isopropyl‐6‐methyl‐4‐pyrimidinyl phosphorothionate) were conducted on Daphnia magna. The 24‐hr static LC50 was 0.86 μL.L‐1. The sublethal effects of 0.05, 0.1, 0.5, 0.75 and 1.0 ngL‐1 of diazinon concentrations on the survival, reproduction and growth of D. magna were monitored for 21 days. The algae Nannochloris oculata (5 x 105 cellsmL‐1) was used to feed the daphnids. The parameters used to determined the effect of the pesticide on D. magna were: mean total young per female; mean brood size; days to first brood; intrinsic rate of natural increase (r); growth; and survival. Reproduction as well as survival was significantly reduced at diazinon concentrations of 0.10 ngL‐1 and higher. The intrinsic rate of natural increase (r) decreased with increasing concentrations of diazinon. Growth, as measured by body length, was depressed significantly at 0.05 ngL‐1 of diazinon and higher concentrations. The maximum acceptable toxicant concentratio...

Chronic toxicity of diazinon to daphnia magna: Effects on survival, reproduction and growth

The sublethal effect of 0.15, 0.18, 0.22, 0.25 and 0.30 μg/L diazinon on the survival, reproduction and growth of D. magna was monitored for 21 days. Neonates (≤ 24 h) were obtained from the stock cultures and raised individually in 50 mL glass beakers. All daphnids were transfered every other day to a new beaker containing fresh medium, food and the appropriate pesticide concentration. The animals were maintained in an environmental chamber at 22±1°C on a 12L: 12D photoperiod and were fed daily on 5 x 105 cell/mL of the green algae Nannochloris oculata. The parameters used to determine the effect of the pesticide on reproduction were: mean total young per female, mean brood size, mean number of broods, mean time to first reproduction and intrinsic rate of natural increase “r”. Growth of surviving adults was determinated after 21 days of exposure. The maximun acceptable toxicant concentration (MATC) was also calculated. Survival was affected with 0.18, 0.25 and 0.30 μg/L diazinon. All concentrations cause...

Chronic toxicity of methylparathion to the rotifer Brachionus calyciflorus fed on Nannochloris oculata and Chlorella pyrenoidosa

The effect of sublethal levels of methylparathion (0,1, 3, 5, 7 mg 1−1) on the freshwater rotifer, Brachionus calyciflorus, during their entire life cycle was studied. Rotifers were fed on two species of unicellular algae: Nannochloris oculata and Chlorella pyrenoidosa; both algal concentrations were 5 × 105 cell ml−1.

Organophosphorus diazinon induced toxicity in the fish anguilla angvilla L.

Abstract 1. Acute toxicity effects of diazinon on European eel ( Anguilla anguilla ) were examined using short-term exposures in static conditions. 2. The lc 50 values found were: 0.16, 0.11, 0.09 and 0.08 mg/1 at 24, 48, 72 and 96 hr exposure, respectively. 3. Eels were exposed to 0.056 mg/l of diazinon and the bioaccumulation and elimination of this insecticide in liver, muscle, gill and blood tissues were studied. 4. BCF were 800 in liver, 1600 in muscle tissue and 2300 and 2730 in gill and blood tissue, respectively. 5. The BCF 1 were 0.30 for liver, 0.60 for muscle and 0.84 for gill. Higher accumulation capacity of the gill was observed for the first hour of exposure. 6. Diazinon elimination from the selected tissues was rapid, diazinon levels were not detected in any tissue after 24 hr in clean water. 7. The excretion rate constants (K 2 ) of this insecticide were 0.023 hr −1 for liver, 0.005 hr −1 for gill and 0.019 −1 for muscle. 8. Diazinon half-lives were calculated as 30.6, 32.2 and 38.3 hr for liver, muscle and gill, respectively.

Uptake and elimination kinetics of a pesticide in the liver of the european EEL

Abstract This study was undertaken to measure the aerobic biodegradation of phenol using poultry litter and to compare the toxicity of the parent compound with the end product(s). Phenol was aerobically treated with a water extract of the untreated litter or of irradiated litter in order to differentiate between the effects of the microorganisms and other chemical species present in the litter on the biodegradation of phenol. Two concentrations of phenol (10 ppm and 100 ppm) were each treated with three concentrations of leachate (0.2%, 0.5% and 1%). The litter leachate (1%) degraded 10 ppm and 100 ppm phenol by 88% in 8 h and 24 h, respectively. Lag phases were observed for all treatments except with 1% leachate. The lag phase increased as the concentration of phenol increased and decreased with an increase in the leachate concentration. No degradation was observed using the leachate of the irradiated litter. Experiments using Ceriodaphnia dubia, (LD50) and Vibrio fischeri, (EC50) showed that the degrada...