Quantification of metallothionein in whole body gammarus locusta (crustacea: amphipoda) using differential pulse polarography

A polarographic method was optimised for the quantitative determination of metallothionein (MT) in whole body Gammarus locusta (Crustacea: Amphipoda). The identity of the MT-like proteins was examined using electrophoretic analysis (SDS-PAGE) and induction studies (water-borne exposures to copper). The MT-method was then used to examine changes in MT levels over the lifetime (juveniles, sub-adults and adults) of a laboratory population of G. locusta; copper (Cu) and zinc (Zn) body burden were also measured by flame atomic absorption spectrometry. MT was quantified by heat-denaturation of 30?000?g supernatants (to remove the bulk of non-MT proteins) followed by differential pulse polarography (DPP) to measure sulphydryl-group containing proteins. DPP proved to be a sensitive and reliable method to quantify heat-stable total sulphydryl-proteins. The majority of the latter was indicated to be MT from SDS-PAGE and induction studies. SDS-PAGE revealed a putative MT of 23?±?0.6?kDa app. mol. wt., consistent with the average size of dimeric MTs (10–20?kDa) found in several aquatic invertebrate species. Copper-exposure revealed parallel increases in the amounts of 23?kDa protein and total sulphydryl-proteins. Levels of whole body MT (rabbit pure MT as standard) range between 1.3 and 2.3?mg?g^?1 dry wt. over the lifetime of G. locusta. Sex-related changes in MT status were observed, the levels in females being 1.6 times higher than in males (p<0.05). Copper and Zn body content changed over the lifetime of G. locusta, the levels in juveniles (p>0.05) and sub-adults (p<0.05) being higher than in adults. It is concluded that the inclusion of MT in ecotoxicological studies with amphipod crustaceans require some caution with respect to the stages of moulting and/or sexual maturation of organisms, but this should not provide any major impediment in its use in metal pollution monitoring.     

Effects of bisphenol A on Gammarus fossarum and Lumbriculus variegatus in artificial indoor streams

The impact of bisphenol A (BPA) on Gammarus fossarum and Lumbriculus variegatus was studied in four artificial indoor streams (0, 5, 50 and 500?µg?L^?1 BPA, nominal) over 103 days in a pulse–dose exposure scenario (weekly BPA application). For G. fossarum populations at day 103, the proportions of juveniles and of breeding females from the highest BPA treatment were in tendency reduced. For individually exposed gammarid pairs an EC₁₀ of 17?µg?L^?1 BPA (nominal) for the proportion of reproductive females in the fourth brood was determined. During the first three broods, the largest brood size occurred at the highest BPA concentration, whereas in the fourth brood it decreased concentration-dependently (fourth brood EC₁₀?=?5?µg?L^?1 BPA, nominal). Effects on L. variegatus were a reduced population growth (103?d-EC₁₀ of 2?µg?L^?1 BPA, nominal) and an increase in dry weight and the number of segments in large, complete worms.     

The correlation of total and extractable heavy metals from soil and domestic sewage sludge and their transfer to maize ( Zea mays L.) plants

A study to understand the mobility and transport of heavy metals (HMs) from soil and soil amended with sewage sludge to maize plants was carried out. The total and ethylenediaminetetraaceticacid (EDTA)-extractable HMs in agricultural soil and untreated domestic sewage sludge samples, and the correlation between the total and extractable metals in soil and sewage sludge were carried out. Pot experiments were performed to study the transfer of HMs to maize grains, grown in soil (control) and in soil amended with sewage sludge (test samples). The total and extractable HMs in soil, sewage sludge, and maize grains were analysed by FAAS/ETAAS (flame atomic absorption spectrometer/electrothermal atomic absorption spectrometer) after digestion in microwave oven. Statistically significant correlations were obtained between the total contents of Cu, Cd, As and their respective extractable fractions in soil, while in domestic wastewater sludge (DWS) the better correlation was observed only for Ni and Cd. The edible part of maize plants (grains) from test samples presented high concentration of Zn, Pb, Ni, Cd, Cu, As, and Cr concentrations (80.7–85.6, 3.8–3.95, 2.35–2.5, 0.75–0.82, 3.21–3.29, 0.23–0.27, and 0.22–0.29?mg?kg^?1, respectively). Good correlations were found between metals in exchangeable fractions of both soil and DWS and total metals in control and test samples of maize grains. The transfer factor of all HMs from DWS to maize grains was also determined.     

Pesticides poisoning cases recorded in Ngaoundere,Cameroon over a 27-month period

An examination of hospital records at Ngaoundere in Cameroon showed that there were 51 reported cases of pesticide poisoning in a 27-month period (January 2004 to April 2006) and of those, only two deaths were attributed to insecticides. One feature was that apart from a high proportion of unidentified poisons, rodenticide poisoning was reported more often than other pesticides. Many cases of poisoning might go unrecorded if an individual does not go to hospital.     

Lead distribution in rats following respiratory exposure to lead-contaminated soils

Two preliminary experiments were performed to explore the use of an intratracheal instillation technique as a cost-effective method of determining the biokinetics of lead (Pb) following respiratory exposure to Pb-contaminated soils. A novel intratracheal instillation procedure was refined and used to deliver a defined dose of Pb-contaminated soil or PbAc to the lower respiratory tract of rats. In the first experiment, rats were sacrificed at numerous time intervals post-dosing, and liver, kidney, blood, and bone tissues were collected for Pb analysis. In the second experiment, rats were dosed with Pb-contaminated soil or PbAc via intratracheal instillation or gastric gavage. All rats were sacrificed 96 h after dose administration and tissues were collected for lead analysis. Data from these experiments indicate Pb is well-absorbed following intratracheal instillation of Pb-contaminated soil, and the intratracheal instillation technique could be used as a cost effective method for exploring the biokinetics of Pb in Pb-contaminated soils following respiratory tract exposure.     

Levels of plasma B-esterases and glutathione-S-transferase activities in three South American toad species

We determined the normal levels of butyrylcholinesterase (BChE), carboxylesterase (CbE), and glutathione S-transferases (GST) activities in three South American toad species in order to establish reference values for field pesticide monitoring purposes. Interspecies variations in B-esterase and GST activities were examined according to body mass. In addition, comparative inhibition of BChE and CbE activities using malaoxon, and chemical reactivation of malaoxon-inhibited BChE activity using pyridine-2-aldoxime methochloride (2-PAM) were investigated. Bufo fernandezae had average activity values for BChE: 17.31 mmol min^?1 ml^?1; CbE: 621.49 nmol min^?1 ml^?1 and GST: 1.94 mmol min^?1 ml^?1 while B. arenarum enzymatic average activities were BChE: 9.51 mmol min^?1 ml^?1; CbE: 270.07 nmol min^?1 ml^?1, and GST: 1.59 mmol min^?1 ml^?1; finally Bufo schneideri had enzymatic mean values of BChE: 2.08 mmol min^?1 ml^?1; CbE: 301.95 nmol min^?1 ml^?1, and GST: 1.60 mmol min^?1 ml^?1. Moreover, we found an allometric relationship between plasma BChE and CbE activities and body size for the three toad species. We suggest that B. fernandezae would be the species with a higher tolerance capacity to organophosphorous insecticides compared to the other toad species, while B. schneideri may be the most vulnerable toad species to field pesticide exposure, although some other factors (e.g., brain AChE sensitivity or pesticide metabolism by phosphotriesterases) should be also taken into account. The malaoxon-inhibited BChE activity of the three toad species was reactivated in the presence of 2-PAM, and it is recommended as a specific and sensitive methodology in the assessment of field exposure to OP insecticides together to compare BChE activity levels between OP-exposed and nonexposed individuals.     

Interaction of alcohol and kola nut on brain sodium pump activity in Wistar rats

The effect of interaction of alcohol and kola nut on sodium pump activity was studied in Wistar rats. Thirty Wistar rats were divided into six groups of five rats per group. Control received a placebo (4 mL of distilled water). Groups 2–6 were treated for a period of 21 days with 10% (v/v) alcohol, 50 mg caffeine/kg, 50 mg kola nut/kg, or a combination of 10% (v/v) alcohol + 50 mg kola nut/kg, or 10% (v/v) alcohol + 50 mg caffeine/kg, respectively. One day after the final exposure, brains were harvested and several biochemical parameters examined including activities of total ATPase, ouabain-insensitive ATPase, ouabain sensitive ATPase (Na⁺–K⁺-ATPase) and levels of nonenzymatic breakdown of ATP and inorganic phosphate (P_i) released. Results showed that the essential enzyme of the brain responsible for neuronal function, Na⁺–K⁺-ATPase, was inhibited by alcohol–kola nut co-administration relative to control, resulting in a decreased ATP production, ion transport and action potential, leading to loss of neuronal activities.     

Measurement of residues of benomyl and mancozeb pesticides in shrub and nonshrub cucumbers sampled from different regions of Mazandaran province (Iran)

Farmers use mancozeb against cucumber pests, such as Perono spoxa, in large amounts without consideration of potential chronic human health hazards resulting from pesticide accumulation in food and fruits. Considering the necessity of food for humans and importance of fruit in daily diet, as well as the determination of pesticide residues in food and fruits as mandated by WHO and FAO, the residues benomyl and mancozeb, the most prominently used pesticides in Iran, were measured. In this study cucumbers from 76 sample regions were used. Data showed that the amounts of benomyl was related to the type of cucumber in the regions, but there was no significant relationship between the amount of mancozeb and type of cucumber.     

Automated indirect fluoride determination by ICP-OES using discontinuous-flow analysis

An automated method for the indirect determination of fluoride by inductively coupled plasma atomic emission spectrometry using discontinuous-flow analysis has been developed. A modified flow injection analysis system was used for sample preparation. The method is based on the determination of excess cerium(III) ion after precipitation of CeF₃. Separation of the precipitate is performed online on a syringe membrane filter that is cleaned each analysis cycle by 1 M nitric acid, ensuring a long life-time for the filter. As a result of the short reaction time, a detection limit of 9.5 mg L^?1 was obtained. The precision (RSD) of the proposed method is 1.02% at fluoride concentration of 28 mg L^?1. The applicability of the technique for groundwater monitoring of fluoride-contaminated area is demonstrated.