Alteration of photosystem II activity by atrazine on Chlamydomonas reinhardtii synchronized and asynchronized cell cycle cultures

Inhibition of photosystem II (PSII) activity by atrazine was investigated in the green alga Chlamydomonas reinhardtii during different states of the cell cycle. The algal cultures were maintained under continuous light or under light/dark cycle (16/8?h) to obtain homogenized cell cycle distribution. The cycle state of algal population was determined by the DNA content using flow cytometry and defined as newly divided cells before the initiation of DNA replication (G₀/G₁) and cells at the end of the replication cycle with fully duplicated DNA content (G₂/M). Under different synchronized states of the cell population, the photosynthetic activity was investigated after treatment at 10, 100, and 1000?µmol?L^?1 atrazine exposed for 24?h by using fluorescence parameters related to PSII activity measured with a plant efficiency analyzer and pulse-amplitude modulated methods. In this study, we found that the atrazine effect was different depending on cell cycle phases and the period of illumination. Algal cells under light–dark cycle showed inhibition of the PSII electron transport leading to an increase of heat energy dissipation by the PSII reaction center. Algal cells grown under continuous light was shown to be more resistant to atrazine than the cells grown under light–dark cycle.     

The role of root organic acids in the tolerance of Festuca rubra to zinc,lead and cadmium

Abstract

Festuca rubra L. plants are pseudometallophytes colonizing abandoned Pb/Zn mine areas, successfully employed in phytostabilization. To study the contribution of low-molecular weight organic acids to metal tolerance, F. rubra plants were grown for three months in hydroponics with Cd (1.8, 18 and 36 µmol?L^?1), Pb (50, 250 and 500?µmol?L^?1) and Zn (0.3, 3 and 6?mmol?L^?1), separately, and in ternary combination (18?µmol?L^?1 Cd + 250?µmol?L^?1 Pb + 0.3?mmol?L^?1 Zn). The roots retained most of the metals but their distribution from shoot to root was altered when the plants were treated with the ternary combination. The main organic acids in roots were citrate and malate. At the lowest concentrations, the metals caused small reductions in biomass, had no effects on photosynthetic pigments nor on malondialdehyde, but led to increases in root organic acids. At higher concentrations, phytotoxic responses were observed, associated with a decline of citrate and malate in the roots.     

Toxic effects of phosphoramidate on Paramecium sp. with special emphasis on respiratory metabolism,growth, and generation time

The toxicological impacts of the increasing number of synthetic compounds present in the aquatic environment are assessed predominantly in laboratory studies where test organisms are exposed to a range of concentrations of single compounds. Protozoan cells are often used as bioindicators for the presence of xenobiotic compounds. In this article, we describe the inhibitory effect of a synthetic phosphoramidate derivative at different concentrations (40, 60, and 80?µmol?L^?1) on Paramecium sp., affecting its growth (proliferation) in concentration-dependent manner, as well as the generation time and response percentage. The LC₅₀ value determined for these protozoa was estimated at 60?µmol?L^?1 on 24?h of exposure. The respiratory metabolism of protozoan is perturbed at three concentrations, noting that the oxygen consumption was significantly increased at high concentrations after 18?h of exposure. In addition, the data can be used as reference values in further testing with other pesticides.     

Differential effects of UV-B radiation fluence rates on growth,photosynthesis, and phosphate metabolism in two cyanobacteria under copper toxicity

This work was undertaken to ascertain the impact of different fluence rates of ultraviolet-B (UV-B) radiation on two cyanobacterial biofertilizers, Phormidium foveolarum and Nostoc muscorum, growing under copper toxicity. Copper (2 and 5?µmol?L^?1) and high UV-B fluence rate (UV-B_H; 1.0?µmol?m^?2?s^?1) decreased the growth, pigment content, photosynthetic oxygen yield, phosphate uptake, and acid phosphatase activity in both the strains analyzed after 24 and 72?h of experiments, and combined exposure further enhanced the toxic effects. Respiration and alkaline phosphatase activities were stimulated appreciably. The damaging effect was shown on the order on pigments: phycocyanin?>?chlorophyll a?>?carotenoids, and on photosystems: whole chain photosynthetic reaction?>?photosystem II?>?photosystem I. Partial recovery in the photosystem II activity in the presence of artificial electron donors; diphenyl carbazide (DPC), hydroxylamine (NH₂OH), and manganese chloride (MnCl₂) pointed out the interruption of electron flow on the oxidation side of photosystem II. Unlike UV-B_H, low UV-B fluence rate (UV-B_L; 0.1?µmol?m^?2?s^?1), rather than causing damaging effect partially, alleviated the toxic effects of Cu. This study suggests that the cyanobacterium P. foveolarum is less sensitive against UV-B_H and excess Cu (2 and 5?µmol?L^?1), thus P. foveolarum may be used as a biofertilizer for sustainable agriculture.     

Producing Cu-loaded algae for feeding experiments: effects of copper on Parachlorella kessleri

Microalgae require several essential metals for optimum growth, which at elevated concentrations may interfere with biochemical and physiological processes, one of them being copper (Cu). The aim of this study is to raise Cu-loaded Parachlorella kessleri as feed for mussels. In order to spike the algae with Cu without lowering their nutritional quality, it is important to know the highest Cu-concentration at which the main parameters remain unaffected, especially in respect to proteins and polysaccharides. The dependence of growth rate, biomass, chlorophyll-a and -b, pheophytin-a, protein, and polysaccharide contents on Cu concentrations are determined. The tests show that P. kessleri is largely unchanged in its nutritional value when exposed to Cu at levels of up to 6?µmol?L^?1. Above 10?µmol?L^?1, toxic effects become obvious, with chlorophyll contents and growth rate being the most sensitive indicators.     

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.     

Effects of copper on lipid peroxidation,glutathione, metallothionein,and antioxidative enzymes in the freshwater mussel Anodonta anatina

Copper is an essential element to all animals. At elevated concentrations, it is toxic and can participate in the formation of reactive oxygen species, leading to cellular damage. In this study, the ecotoxicological relevance of copper was investigated with freshwater mussels, Anodonta anatina. When the mussels were exposed to copper at environmentally realistic concentrations, either via the water (0.3?µmol?L^?1 Cu) or fed with Cu-loaded algae (equivalent to 0.06?µmol?L^?1 Cu), the level of thiobarbituric acid-reactive substances rose and glutathione decreased. This was associated with the induction of metallothionein and, relative to total protein, of glutathione reductase and the antioxidative enzymes superoxide dismutase, catalase, and glutathione peroxidase. But, since the overall protein-synthetic capacity was hampered by the copper insult, the activities of the enzymes relative to tissue weight and copper concentrations were depressed. During depuration, most parameters started to normalize although not returning to control values within 12 days.     

In vitro kinetic characterization of inhibition of acetylcholinesterase by organophosphate and carbamate compounds in food animals

Organophosphate, dichlorvos (2,2-dichlorovinyldimethylphosphate, DDVP), and diazinon (DZN) as well as carbamates are currently found in contaminated water, dust, soil, agricultural waste, and elsewhere in the environment, and can be harmful after accidental or deliberate exposure. Intoxication by these compounds causes a generalized cholinergic crisis due to the inhibition of acetylcholinesterase (AChE), whose major physiological role in mammalian tissues is in nervous transmission. The half-maximal inhibitory concentration (IC₅₀) was found to be higher for carbaryl than DDVP and DZN, about 3–4 times in liver and about 4–6 times in muscles. The half time (t _1/2) of inhibition (8?µmol?L^?1 DDVP or DZN; 40?µmol?L^?1 carbaryl) ranged between 4 and 16?min and decreased as follows: pig?>?cattle?>?sheep for liver tissue, and cattle?>?sheep?>?pig for muscle. Rate constants of inhibition (k _i) ranged between 43?×?10^?3 and 168?×?10^?3?min^?1 for liver and from 46?×?10^?3 to 115?×?10^?3?min^?1 for muscles. Very little residual AChE activity (<6.1%) was seen in liver, but more was seen in muscle (<17%).     

Seasonal changes in inorganic nutrient concentration in the alexandria western harbour (Egypt)

Surface and bottom water samples were collected on a monthly basis from six locations in the Alexandria Western Harbour between April 2002 and March 2003. Total nitrogen, ammonia, nitrite, nitrate, total phosphorus, reactive phosphate and silicate were analysed. The average content of total nitrogen ranged from 81.1 to 65.7?µmol?l^?1 in the surface and bottom waters, respectively, while ammonia ranged from 13.77 to 15.79?µmol?l^?1 in surface and bottom waters, respectively. Also, the average concentration of nitrite was relatively higher in surface waters than in bottom waters (0.89 and 0.61?µmol?l^?1, respectively). The results of this study also indicated a considerable temporal variation in nitrate concentrations which ranged from 1.12 to 13.83?µmol?l^?1. Total phosphorus displayed an irregular pattern throughout the year, ranging from 1.9 to 11.8?µmol?l^?1 in surface waters and from 1.7 to 9.1?µmol?l^?1 in bottom waters. The results of PO₄-P analysis showed higher values in surface waters (0.28–2.75?µmol?l^?1) than in bottom waters (0.10–1.70?µmol?l^?1). The average concentration of silicates was relatively lower in the surface than in the bottom waters (8.97 and 10.1?µmol?l^?1, respectively). The analysis of variance (ANOVA) among seasons and sites revealed significant differences for ammonia, total nitrogen and phosphate, while nitrate showed no significant differences among stations. Finally, silicate did not show any significant variance among sites and seasons (ANOVA, P?>?0.05).     

Induction of micronuclei in tadpoles of Odontophrynus americanus (Amphibia: Leptodactylidae) by the pyrethroid insecticide cypermethrin

Cypermethrin (CY) is an active cyano pyrethroid effective against a wide range of pests encountered in agriculture and forestry. Although CY is not mutagenic in in vitro assays for gene mutation, in vivo assays showed conflicting results. In vivo genotoxicity of the synthetic pyrethroid CY in erythrocytes of Odontophrynus americanus tadpoles was examined. The frequency of micronuclei (MN) was recorded in blood smears obtained from tadpoles exposed in vivo to four different nominal concentrations 5, 10, 20 or 40?µg?L^?1 of the compound and fixed at two sampling times 48 and 96?h. As a positive control larvae were exposed to 40?mg?L^?1 of cyclophosphamide (CP). Tadpoles exposed to all CY treatments showed a significant increase in single small MN compared to the negative control group after 48?h and at 5 and 10?µg?L^?1 of CY at 96?h. Results obtained here demonstrated the genotoxic effects of the commercial formulation CY in the anuran larvae analyzed. Thus, data suggest that measurements of MN and other erythrocytes morphological aberrations performed in circulating blood samples of O. americanus tadpoles is a method for detecting cytogenetic damage in other native species.     

Deltamethrin-induced alterations in serum calcium and prolactin cells of a freshwater teleost,Heteropneustes fossilis

Deltamethrin, a pyrethroid insecticide widely used in many countries, causes significant adverse effects in aquatic ecosystems. The concentration of deltamethrin in water reservoirs and the run off from agricultural areas (in water) in many countries range up to 24.0?µg?L^?1 which is higher than the recommendation of the European Union standard. Hence, in this study the effects of deltamethrin were investigated, i.e. its toxic impacts on the freshwater catfish Heteropneustes fossilis in terms of serum calcium and prolactin cells (located in the rostral pars distalis region of the pituitary). The fish were subjected to deltamethrin for a short-term experiment (96?h; 1.5?µg?L^?1 e.0.8 of 96?h LC₅₀) and a long-term experiment (28 days; 0.37?µg?L^?1 e.0.2 of 96?h LC₅₀). After short-term deltamethrin exposure, serum calcium levels decrease. No histological change in prolactin cells is noticed throughout the short-term experiment. Fish exposed to deltamethrin for 7 days also exhibit a decrease in serum calcium level. This decrease persisted until the end of the experiment (28 days). Prolactin cells of fish treated for 14 days with deltamethrin exhibit increased nuclear volume and degranulation, increasing progressively from the 21st day onwards. After 28 days, a few degenerated cells are discerned. The results of this study show that deltamethrin is moderately toxic for the freshwater fish H. fossilis by producing adverse effects on serum calcium and prolactin cells. Hence, it should be used with caution in areas near fish-inhabited waters.     

Impaired phospholipid metabolism in Saccharomyces cerevisiae by increased phospholipase B activity induced by 4-(methyl nitrosamino)-1-(3-pyridyl)-1-butanone

Tobacco products and cigarette smoke cause many respiratory diseases including cancer. 4-(Methyl nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is a potent carcinogen in cigarette smoke, but its effect on lipid metabolism remains enigmatic. Hence, Saccharomyces cerevisiae was exposed to different concentrations of NNK (0–400?µmol?L^?1) to elucidate its role in lipid metabolism. Exposure to NNK substantially decreases (about 60%) of the phospholipid content with a concomitant increase in lysophospholipids. Significant reduction was observed in the phosphatidylcholine followed by phosphatidylethanolamine with NNK-treated cells. On the contrary, cells accumulated significant amount of neutral lipids and free fatty acids. Exposure of yeast cells (wild-type cells and three plbΔ mutant strains) to NNK greatly enhances the hydrolysis of phospholipid in the presence of calcium. We are the first to report that exposure to NNK enhances phospholipase B (PLB), particularly plb1p activity. Furthermore, NNK also promotes the alteration of phospholipid fatty acid (FA) content. These results suggest that NNK aids in the degradation of phospholipids by enhanced PLB activity and is accompanied with FA alteration. Understanding the altered phospholipid metabolism in the presence of NNK remains a worthy pursuit.     

Effects of chronic bisphenol A exposure on hepatic antioxidant parameters in medaka (Oryzias latipes)

Bisphenol A (BPA), the plastic monomer and plasticizer with well-known endocrine-disrupting chemical properties, is widely present in the aquatic environment, but little is known regarding hepatic toxicity in fish. This study assessed the effects of chronic exposure to BPA on the antioxidant defense system of Japanese medaka liver. Fish were exposed to sub-lethal concentrations of BPA (0.1, 1, 10, 100, or 1000?µg?L^?1) for 60 days. Antioxidant parameters were determined and compared with controls. The results showed that the activities of catalase (CAT), superoxide dismutase, glutathione peroxidase, glutathione S-transferase, and the content of reduced glutathione were all induced at 10?µg?L^?1 BPA after 60 days. Further, hepatic CAT was sensitive to BPA at 1?µg?L^?1 concentration. In conclusion, CAT may serve as a potential early biomarker of BPA exposure in aquatic environment.     

Efficiency of Zn phytoextraction,biomass yield and formation of low-molecular-weight organic acids in S × rubens – a hydroponic experiment

The aim of the study was to estimate Zn phytoextraction and changes in biomass of S?×?rubens growing in modified Knop's solution with different levels of Zn addition (0.5, 1.0, 2.5 and 5.0?mM). Obtained results were correlated with secretion of selected low-molecular-weight organic acids (LMWOAs) in the rhizosphere, roots and leaves. An increase in Zn concentration in Knop's solution resulted in Zn accumulation in roots, shoots and leaves. The highest accumulation was observed for plants growing in 5?mM Zn, at concentration levels 4741.36?±?98.66, 1227.31?±?16.57 and 2241.65?±?34.90?mg?kg^?1 DW in roots, shoots and leaves, respectively. The bioaccumulation factor and the translocation factor for plants growing in 0.5, 1.0 and 2.5?mM Zn clearly indicate that this Salix taxon is an effective Zn accumulator. The general reduction of Salix biomass with an increase in Zn concentration in the solution was observed. In the rhizosphere, the total LMWOA concentration was almost 0.93?µmol?kg^?1 DM for control (Zn free) plants, while for 5.0?mM of Zn it was 4.9?µmol?kg^?1 DM. Increasing concentrations of acids were observed in roots (1.34 for the control and 5.57?µmol?kg^?1 DM for plants treated with 2.5?mM of Zn).     

Green chemistry approach to analysis of formic acid and acetic acid in aquatic environment by headspace water-based liquid-phase microextraction and high-performance liquid chromatography

A simple and totally organic-free (green) method, viz. headspace water-based liquid-phase microextraction combined with high-performance liquid chromatography-ultraviolet detection has been successfully developed for analysis of formic acid and acetic acid in environmental water samples. A microdrop of an aqueous solution of sodium hydroxide was suspended from the tip of a microsyringe needle over the headspace of the stirred sample solution containing the analytes at pH 1.0 for a given time. The microdrop was then retracted into the microsyringe, diluted with HPLC mobile phase, and injected to HPLC. Optimum efficiency has been achieved for: 3.0 µL NaOH microdrop (0.1 mol L^?1) exposed for 15 min over the headspace of an aqueous sample of 6.5 mL at 55 °C, containing 15% w/v of Na₂SO₄, adjusted to pH = 1.0 and stirred at 750 rpm. Under these conditions, enrichment factors of 162 and 187, limits of detection of 0.3 and 0.1 µg L^?1 (S/N = 3) with dynamic linear ranges of 1–500 and 0.5–500 µg L^?1 were obtained for formic acid and acetic acid, respectively. A reasonable repeatability (5.8% ≤ RSD ≤ 8.8%, n = 6) and satisfactory linearity (r² ≥ 0.997) illustrated the performance of the method.     

Combined effects of copper and cadmium on Chlorella pyrenoidosa H.Chick: subcellular accumulation,distribution, and growth inhibition

Disposal of waste into aquatic ecosystems may cause microalgae to be exposed to various metals, e.g. copper and cadmium. The effects caused by combinations of metals may be more serious. Evaluations of subcellular fate, bioaccumulation, and biological effects of metals on aquatic organisms are generally derived from experiments with individual metals. The present study aims to evaluate the effects of exposure of Chlorella pyrenoidosa to copper and cadmium in combination on subcellular accumulation, distribution, and growth. The algae were exposed for 72 h to copper at concentrations of 13 – 25 µmol L^?1, cadmium at about 6 µmol L^?1, and combinations thereof. The levels of copper and cadmium in subcellular organelles, heat-denaturated protein, metal-rich granules, and heat-stable protein were determined by atomic absorption spectrometry. Exposure of C. pyrenoidosa to copper and cadmium in combination inhibited growth more strongly than copper and cadmium individually. Highest accumulation was observed in metal-rich granules and heat-stable proteins. Administration of both metals in combination affected their subcellular distribution: copper was mainly distributed into the metal-rich granules (70%–80%) and heat-stable proteins (9%–24%), cadmium in the metal-rich granules (88%–98%).     

Formation of resistance in the Chironomus plumosus to four pesticides over 45 generations

A study was conducted on the Chironomus plumosus larvae to determine initiation of resistance to four pesticides – chlorfenvinphos, chlorpyrifos, chlorpyrifos-methyl, and malathion. First generational LC₅₀ values were well within the threshold value for chironomids based upon the literature. Subsequent LC₅₀ values were observed to increase, indicating a lessening of the toxicity of the pesticides to the chironomid. In the case of chlorfenvinphos, the 96 h LC₅₀ for generations 1–23 was 6 µg L^?1, in generations 3–7 was 8.57 µg L^?1 and 11.14 µg L^?1 for generations 8–9. Generations 10–12 had an LC₅₀ value of 22.58 µg L^?1 and generation 13 had an LC₅₀ value of 35.08 µg L^?1. Generation 14 had an LC₅₀ value of 47.58 µg L^?1. Generations 15–19 and 20–24 had 96 h LC₅₀ values of 60.68 µg L^?1, 72.58 µg L^?1, 85.08 µg L^?1, 97.58 µg L^?1 and 110.08 µg L^?1, respectively. Generations 25, 26 to 30, 31 to 38 and 39 to 45 had 96 h LC₅₀ values of 160.42 µg L^?1, 210.7 µg L^?1, 262.24 µg L^?1 and 274.36 µg L^?1, respectively. The variation between LC₅₀ values was found to be statistically significant. This was observed for most pesticides tested. Larval size and life cycle duration was observed to change from generation to generation with the body size decreasing markedly from 1 to 0.3 cm with life cycle increasing from 7 to 39 days.     

In situ seagrass photosynthesis measured using a submersible, pulse-amplitude modulated fluorometer

Assessments of photosynthetic activity in marine plants can now be made in situ using a newly developed, submersible, pulse-amplitude modulated (PAM) fluorometer: Diving-PAM. PAM fluorometry provides a measure of chlorophyll a fluorescence using rapid-light curves in which the electron-transport rate can be determined for plants exposed to ambient light conditions. This technique was used to compare the photosynthetic responses of seagrasses near Rottnest Island, Western Australia. Several fluorescence parameters were measured as a function of time of day and water depth; electron-transport rate (ETR), quantum yield, photochemical quenching and non-photochemical quenching and Photosystem II (PSII) photochemical efficiency (F _v :F _m ratio) were measured. Results indicate that recent light-history plays a crucial role in seagrass photosynthetic responses. Maximum ETR of Posidonia australis, Amphibolis antarctica and Halophila ovalis is influenced by the irradiance during the diurnal cycle, with low rates at dawn and dusk (<10 μmol electron m⁻² s⁻¹), highest rates in late morning (40 to 60 μmol electron m⁻² s⁻¹) and a mid-day depression. Maximum ETR and PSII photochemical efficiency varied widely between seagrass species and were not correlated. A comparison of photochemical to non-photochemical quenching indicated that seagrasses in shallow water receiving high light have a high capacity for non-photochemical quenching (e.g. light protection) compared to seagrasses in deep water. These results indicate that in situ measurements of photosynthesis will provide new insights into the mechanisms and adaptive responses of marine plants. Received: 26 May 1997 / Accepted: 27 May 1998     

Effects of sublethal concentrations of permethrin on bioaccumulation of cadmium in zebra cichlid (Cichlasoma nigrofasciatum)

The aim of this study was to evaluate the synergistic and antagonistic effects of permethrin on the bioaccumulation of cadmium in zebra cichlid (Cichlasoma nigrofasciatum). Zebra cichlid was exposed to various sublethal concentrations of permethrin combined with cadmium chloride for 15 days. Although at the lowest concentrations, permethrin (0.40 µg L^?1) decreased the bioconcentration potential of cadmium in fish, bioaccumulation of cadmium increased along with increasing concentrations of permethrin (0.80–1.20 µg L^?1). However, the magnitude of changes in the accumulation of cadmium in the fishes simultaneously exposed to 1.60 µg L^?1 permetrin and cadmium chloride was similar to the cadmium levels observed in fishes after exposure to the cadmium chloride alone. The results of this study suggest that the synergistic effects of permethrin on the bioaccumulation of cadmium highly depend on the concentrations of this pesticide in water.     

Arsenic in freshwater ecosystems of the Bengal delta: status,sources and seasonal variability

The study aimed to examine the contamination status of arsenic (As) in excavated small water bodies, commonly known as ponds – the integral part of daily life in the arsenic-affected rural areas of West Bengal, India in comparison to the unaffected areas. The ponds of the contaminated area had higher levels of As: water 2–174 µg L^?1 (mean 31 ± 2 µg L^?1) and sediment 1.3–37.3 mg kg^?1 (mean 10.3 ± 0.4 mg kg^?1), than those from the unaffected area: water 1–8 µg L^?1 (mean 4 ± 0 µg L^?1) and sediment 1.4–5.3 mg kg^?1 (mean 3.0 ± 0.1 mg kg^?1). A moderate positive correlation was observed between the water and sediment arsenic content of the ponds of the arsenic-affected region (r = 0.688, n = 277, p < 0.0001). Contaminated ground water, either as direct input or through agricultural washings, was found to be the major contributor of arsenic pollution to these ecosystems. Seasonal variations were not prominent. This study emphasized the beneficial role of using the studied ecosystems over the highly contaminated ground water for various livelihood activities in the Gangetic delta region.