Toxicity of the nerve agent tabun to Daphnia magna,a new experimental species in military toxicology

Daphnia magna, a freshwater microcrustacean, is currently tested as an alternative experimental species in research dealing with nerve agents poisonings treatment. Because of this, the toxicity of the nerve agent tabun (a cyanide-group containing organophosphate) to Daphnia had to be examined by estimating the EC₅₀ values. The immobilization of daphnids was chosen as the end-point. It was found that D. magna is sensitive to small amounts of tabun, even after 15 min exposure, and tabun toxicity increases with time. The estimated EC₅₀ values for 15, 30, 45, and 60 min exposure were as follows: 67.39, 38.10, 26.95, and 21.9 μg l^?1. In addition, the toxicity of media to which tabun was added 24 h before the start of experiments was examined. The results obtained indicate that daphnids can be used in experiments with nerve-agent intoxication treatment.     

Movements of tiger sharks (Galeocerdo cuvier) in coastal Hawaiian waters

The giant clam Tridacna crocea harbors in the mantle tissue symbiotic microalgae commonly called zooxanthellae. Isolated zooxanthellae release glycerol into the medium in the presence of mantle tissue homogenate (MH), but it is not clear whether the cells do so in situ. In order to determine the photosynthetic products released by zooxanthellae in the mantle of the giant clam we traced photosynthetic fixation products from ¹³C- and ¹⁴C-bicarbonate both in the clam and in isolated zooxanthellae (IZ) in the presence or absence of MH. After 15 min incubation in the absence of MH the IZ released less than 0.6% of the fixed labeled carbon, mainly as glucose. The major intracellular photosynthates were neutral lipids, which constituted 20 to 40% of the total extractable ¹⁴C. In the presence of MH, the IZ released up to 5.6% of the total fixed ¹⁴C, mostly as glycerol, and the major intracellular photosynthate was glucose. In an intact clam incubated in sea water containing ¹⁴C-bicarbonate, 46 to 80% of the fixed ¹⁴C was translocated from the zooxanthellae to the host tissues. Most of the ¹⁴C in the hemolymph, in the isolated zooxanthellae and in intact mantle tissue (containing zooxanthellae) was recovered as glucose. No ¹⁴C-glycerol was detected in the mantle after 1 to 30 min incubation, and, even after 60 min, far less ¹⁴C-glycerol was synthesized than by IZ in the presence of MH. The possibility that in clam tissue glycerol is converted to glucose was examined by tracing the labeled carbon from ¹⁴C-glycerol injected into the adductor muscle. After 5 min incubation, no labeled glucose was found in the hemolymph, but after 60 min, some 20% was found as glucose. Thin slices containing zooxanthellae, cut from the surface of the mantle, fixed inorganic carbon supplied as NaH¹⁴CO₃ in the medium and mainly released ¹⁴C-glucose. The addition of MH to the surrounding medium did not affect the release rate or form of release product. When the slices were cut into smaller pieces, however, the ratio of glycerol to glucose in the release product increased. These results indicate that in the presence of MH the metabolism of isolated zooxan- thellae was different from that of zooxanthellae in the mantle. In the presence of MH, isolated zooxanthellae release mostly glycerol, whereas in the mantle they release glucose. Received: 18 February 1998 / Accepted: 4 December 1998     

The pattern of carbon fixation in the marine unicellular alga Phaeodactylum tricornutum

The short- and long-term fixation of ¹⁴CO₂ by Phaeodactylum tricornutum was studied using methods of fractionation that allowed examination of all the products labelled with ¹⁴C. There was no doubt that the major pathway of CO₂ fixation was into 3-phosphoglycerate, but there was also significant incorporation by -carboxylation by means of phosphoenol-pyruvate carboxylase and transamination into aspartate. At short time intervals (10 sec), 90% of the radioactive products found were accounted for by 3-phosphoglycerate and aspartate. The lipids associated with the photosynthetic apparatus contained a high proportion of the ¹⁴C fixed, which at 60 sec was located mainly in the carbohydrate portion of the lipids. At 30 and 300 min, the chlorophylls, carotenoids and the long-chain fatty acids were heavily labelled with ¹⁴C. The monogalacto-diglycerides, the digalacto-diglycerides and the sulpholipids each had a characteristic long-chain fatty acid composition. The cell proteins and a reserve polysaccharide were also labelled with ¹⁴C at short time intervals and increased their radioactivity in a linear fashion up to the longest period studied, 300 min. The activity of the enzymes ribulose diphosphate carboxylase and phosphoenolpyruvate carboxylase was sufficient to account for the pattern of fixation found.     

Photochemistry of halogenated benzene derivatives. part xii.∗ effects of sodium nitrite on the environmental phototransformation of bromoxynil (3,5‐dibromo‐4‐hydroxybenzonitrile) herbicide in water: The photoincorporation of nitrite ions into bromoxynil

Photoreactions of the herbicide bromoxynil (3,5‐dibromo‐4‐hydroxybenzonitrile) (1) were extensively studied in water containing various concentrations of sodium nitrite with radiation of wavelengths around 313 nm. In the absence of NaNO₂, the quantum yields for the photodecomposition of the herbicide I amounted to 0.054±0.005, while such data was in the range of 0.047 ±0.005 to 0.023 ±0.003 in the presence of 0.5 to 25.0 × 10^‐3 m of the inorganic salt. These quantum yield data for the phototransformation of 1 without and with the presence of sodium nitrite followed the Stern‐Volmer equation. The rates of the photolytic destruction of bromoxynil (1) in water were slower in the presence than in the absence of NaNO₂. For example, the irradiation of the 7.8 x 10^‐6M aqueous solution of 1 in the presence of 10 mM sodium nitrite gave rise to the production of three main photoproducts, viz., 3‐bromo‐4‐hydroxy‐5‐nitrobenzonitrile (2), 4‐hydroxy‐3‐nitrobenzonitrile (3), and an “unidentified photoproduct”; 4. In the case of this photoreaction of compound 1, the percentages of maximum concentration of the photoproducts 2, 3, and 4 were achieved after 14, 44.5, and 60‐min photolyses of 1, respectively; the starting material 1 completely disappeared after 60‐min photolysis. The photoproducts 2 and 3 were identified through the interpretation of the GC‐MS data. Both thhe mass and FT‐IR spectra of the product 4 indicated the possible presence of a COOH group in 4.     

Ammonia production and kinetic properties of glutamate dehydrogenase in the sipinculid Phascolosoma arcuatum exposed to anoxia

The amounts of total NH ₄ ⁺ detected in the external media in which Phascolosoma arcuatum had been exposed to various periods of anoxia were significantly greater than those in which the worms were exposed to normoxia for a similar period. The increased NH ₄ ⁺ production by P. arcuatum during anoxic exposure was unlikely to be due to an increased catabolism of adenine nucleotides or urea. In contrast, there were significant decreases in the concentrations of several free amino acids in the coelomic plasma and body tissues of individuals during the 48 h of anoxic exposure. The amount of NH ₄ ⁺ produced by the anoxic P. arcuatum could be accounted for by the decreases in the concentrations of aspartate or glycine. Increases in the catabolism of free amino acids (FAA), leading to the increased production of NH ₄ ⁺ , in P. arcuatum during anoxia were supported by the detection of significant changes in the kinetic properties of glutamate dehydrogenase (GDH), in the deaminating direction, from worms exposed to anoxia for 48 h. The apparent increase in the affinity of GDH from the anoxic worm to glutamate would bring about a greater deaminating activity at physiological concentrations of ths substrate. P. arcuatum used in these experiments were collected from the mangrove swamp at Mandai, Singapore between 1990 and 1993.     

Effects of arsenic on survival and metabolism of Crangon crangon

Metabolic changes and toxic effects in Crangon crangon (collected during summer 1989 near Helsingør, Denmark and at Kulhuse in the Isefjord) were observed during exposure to different arsenate concentrations. Survival, respiration, and concentrations of hemolymph glucose, muscle glycogen andadenine nucleotides were measured. C. crangon was very tolerant of arsenate; survival was only affected at arsenate concentrations >25 ppm. Small individuals (0.2 to 0.29 g wet weight) were less tolerant than medium-sized and large individuals (0.5 to 0.59 g and 0.8 to 0.89 g wet weight), e.g. at 50 ppm the LT₅₀ for small individuals was 180 h compared to 343 h and 360 h in middle-sized and large individuals. The relation between size and toxicity is probably due to the change in surface/volume ratio. Under clean conditions the respiration of C. crangon showed a clear relation between routine MO₂ (O₂ in mlg^-1 h^-1) and wet weight (r²=0.89; P<0.001). During shortterm exposure to arsenate (10 and 50 ppm), the respiratory rates (MO₂) seemed to decrease in a size-dependent manner. The concentrations of blood glucose and muscle glycogen did not change in response to arsenate exposure. The concentrations of adenylate phosphates were high and constant throughout the experiments. C. crangon had maximal adenylate energy charge (AEC) values of ca. 0.8. No decrease in AEC due to the exposure to arsenate was observed.     

Synthesis and antibacterial activity of a Fe3O4–AgCl nanocomposite against Escherichia coli

In this investigation, Fe₃O₄ magnetic nanoparticles (MNPs) were prepared by the alkalinization of an aqueous medium containing ferrous sulfate and ferric chloride. In the next step, a Fe₃O₄–AgCl magnetic nanocomposite was fabricated by the drop-by-drop addition of silver nitrate solution into a NaCl solution containing Fe₃O₄ MNPs. All prepared nanoparticles were characterized by transition electron microscopy (TEM), X-ray diffraction (XRD), and energy-dispersive X-ray spectroscopy (EDS). Both particle types varied in size from 2.5 to 20?nm, with an average size of 7.5?nm for Fe₃O₄ MNPs and 12.5?nm for Fe₃O₄–AgCl nanocomposites. The antibacterial effect of the Fe₃O₄ MNPs and fabricated Fe₃O₄–AgCl nanocomposites against Escherichia coli (ATCC 35218) were investigated by conventional serial agar dilution method using the Müller–Hinton Agar medium. The minimum inhibitory concentration was 4?mg?mL^?1 for Fe₃O₄ MNPs and 2?mg?mL^?1 for the Fe₃O₄–AgCl magnetic nanocomposites. Time-kill course assays showed that the Fe₃O₄–AgCl magnetic nanocomposites successfully killed all inoculated bacterial cells during an exposure time of 60?min. The antibacterial activity of recycled Fe₃O₄–AgCl magnetic nanocomposites over four 60?min cycles of antibacterial treatment was further tested against E. coli by the colony-forming unit (CFU) method. The antibacterial efficiency of the nanocomposites was constant over two cycles of antibacterial testing.     

Effects of light exposure on the retention of kleptoplastic photosynthetic activity in the sacoglossan mollusc <Emphasis Type="Italic">Elysia viridis</Emphasis>

The effects of light exposure on the photosynthetic activity of kleptoplasts were studied in the sacoglossan mollusc Elysia viridis. The photosynthetic activity of ingested chloroplasts was assessed in vivo by non-destructively measuring photophysiological parameters using pulse amplitude modulation (PAM) fluorometry. Animals kept under starvation were exposed to two contrasting light conditions, 30 μmol photons m⁻² s⁻¹ (low light, LL), and 140 μmol photons m⁻² s⁻¹ (high light, HL), and changes in photosynthetic activity were monitored by measuring the maximum quantum yield of photosystem II (PSII), F _v/F _m, the minimum fluorescence, F _o, related to chlorophyll a content, and by measuring rapid light-response curves (RLC) of relative electron transport rate (rETR). RLCs were characterised by the initial slope of the curve, α_RLC, related to efficiency of light capture, and the maximum rETR level, rETR_m,RLC, determined by the carbon-fixation metabolism. Starvation induced the decrease of all photophysiological parameters. However, the retention of photosynthetic activity (number of days for F _v/F _m > 0), as well as the rate and the patterns of its decrease over time, varied markedly with light exposure. Under HL conditions, a rapid, exponential decrease was observed for F _v/F _m, α_RLC and rETR_m,RLC, F _o not showing any consistent trend of variation, and retention times ranged between 6 and 15 days. These results suggested that the retention of chloroplast functionality is limited by photoinactivation of PSII reaction center protein D1. In contrast, under LL conditions, a slower decrease in all parameters was found, with retention times varying from 15 to 57 days. F _v/F _m, α_RLC and rETR_m,RLC exhibited a bi-phasic pattern composed by a long phase of slow decrease in values followed by a rapid decline, whilst F _o decayed exponentially. These results were interpreted as resulting from lower rates of D1 photoinactivation under low light and from the gradual decrease in carbon provided by photosynthesis due to reduction of functional photosynthetic units.     

Respiratory impairment by water-borne copper and zinc in the edible crab cancer pagurus (L.) (Crustacea: Decapoda) during hypoxic exposure

Specimens of the edible crab Cancer pagurus (L.) collected in the Skagerrak, Denmark, between June and August 1990 were examined in the laboratory. Impairment of respiratory function after pre-exposure (7 d) to sub-lethal concentrations of Cu and Zn (0.4 mg l^-1) was only detectable during hypoxic exposure [PO₂ (partial pressure of oxygen) =60 torr]. This was indicated by a decrease in the transfer factor (TO₂), due principally to an increase in the PO₂ differential across the gills. Cu and Zn exposure did not cause significant changes in ventilation or perfusion although there was some indication that cardiac output may increase in respiratory-impaired individuals. After 28 d exposure no difference was noted in the respiratory responses to hypoxia of treated and untreated crabs. It is concluded that respiratory impairment was due to an increase in the diffusion barrier thickness at the gills and that this was reversible even during continued exposure to trace metal contamination.     

Carbon fixation and analysis of assimilates in a coral-dinoflagellate symbiosis

Freshly collected pieces of the hermatypic coral Acropora cf. scandens containing dinoflagellate endosymbionts (presumably Gymnodinium microadriaticum) were allowed to assimilated ¹⁴C from H¹⁴CO ₃ ^- in the light and in the dark. Time-dependent carbon uptake resulted in intense ¹⁴C-labelling of ethanol-soluble as well as of insoluble assimilates. About forty ¹⁴C-labelled assimilates have been identified. Polymeric (ethanol-insoluble) compounds achieve about 30% of total radiocarbon incorporation after 60 min incubation. Kinetics of ¹⁴C-labelling of single assimilates are analyzed. Percentages of typical photosynthates in the soluble fraction undergo characteristic time-dependent changes. Lipids proved to be the main accumulation products of carbon assimilation by incorporating more than 50% of ¹⁴C after 60 min photosynthesis. The data indicate that low-molecular weight photosynthates such as ¹⁴C-glycerol and ¹⁴C-glucose are rapidly converted to constituents of the polymeric fraction(s) of the coral. Besides peptides, polysaccharides, and lipophilic substances, considerable amounts of ¹⁴C are confined to skeletal CaCO₃ of the coral. The results are discussed with respect to trophic and metabolic interrelationships between the autotrophic dinoflagellates and the A. cf. scandens tissues.     

Experimental study on acute effects of the combined exposure to temperature increase and chlorination upon the marine copepod Acartia omorii

The acute effects of combined exposure to temperature increase and chlorination on a neritic marine copepod, Acartia omorii Bradford (collected offshore of Onjuku, Japan in 1982), were investigated in the laboratory. Continuous flow exposure and batch exposure modes were compared. Based on the results of continuous flow experiments, the 24-h median lethal concentration (24-h LC₅₀, in mg l^-1) of total residual chlorine was estimated using the multiple regression equation below, with a multiple correlation coefficient of 0.955: 24-h LC₅₀=2.988-0.034 dT-1.611 log₁₀ t where dT is temperature rise (°C) and t is exposure duration (min). In batch experiments, the predictive power of the multiple regression equation was reduced, probably due to variations in chlorine concentration during exposure duration.     

The short-term partitioning of carbon-14 assimilate between zooxanthellae and polyp tissue in Acropora formosa

The usefulness of Fluorinert for the extraction of Acropora formosa polyp tissue and zooxanthellae was demonstrated. The latter remain intact, with no leakage of metabolites, and the polyp tissue can be extracted in a minimal volume. Intact A. formosa and its isolated zooxanthellae were incubated in the light with sodium [¹⁴C]bicarbonate for 5 s to 15 min and the kinetics of carbon-14 fixation was determined. The isolated zooxanthellae showed a linear response for carbon fixation, whilst the zooxanthellae in the intact association showed a lag period of 1 to 2 min, containing only 12% of the total fixed carbon in the first 1 min. After 10 min, the distribution of fixed carbon between the symbiotic partners was approximately even and the total carbon fixed was in a range similar to that fixed by the isolated zooxanthellae. A pulse-chase experiment showed rapid movement of fixed carbon from the polyp tissue to the zooxanthellae after the 30 s pulse. The paper discusses two possible explanations for the observed results.     

Anaerobic metabolism in Arenicola marina and Nereis diversicolor during low tide

In order to observe the metabolic response of two marine polychaetes, Arenicola marina L. and Nereis diversicolor Müller, at various times during low tide, the concentrations of adenine nucleotides, phosphagens, D- and L-alanine, succinate, acetate, propionate and lactate were measured. During 8h of exposure ATP does not change significantly in the body-wall musculature of the lug-worm A. marina, whereas ADP and AMP concentrations double the first hour of exposure. Correspondingly, the energy charge falls from 0.91 to about 0.84. The concentration of the phosphagen phosphotaurocyamine decreases by about 20% during the first 2 h and after 6 h only about 50% of the initial concentration was measured. During exposure, aspartate decreases by approximately 6 mol g^-1 fresh weight within 6 h (from 23 to 17 mol g^-1 fresh weight). During the first 4 h of exposure succinate concentration doubles and within the next 4 h it reaches values about 10-fold the initial value. The concentrations of acetate and propionate only increase during the last 2 h (6–8) of exposure. After several hours of exposure typical changes in the concentrations of all these metabolites could also be demonstrated in N. diversicolor. The concentrations of the two phosphagens phosphoglycocyamine and phosphocreatine and of aspartate decrease by about 25% during 9 h of exposure, on the other hand succinate and especially D-lactate accumulate. In both species, the partial anaerobic energy production during exposure does not have a lasting detrimental effect. After one high tide the worms have fully recovered.     

Influence of environmental salinity on oxygen consumption and ammonia excretion of the arctic under-ice amphipodOnisimus glacialis

Changes in salinity affect the metabolic rate of the sympagic amphipodOnisimus glacialis collected from the Barents Sea in 1986 and 1988. When transferred from 35 to 5 ppt S, oxygen consumption and ammonia excretion both increase three-fold during the first 5 h of exposure, and they remain high throughout the rest of the experimental period (26 h). During 24-h acclimation to various salinities (5 to 45 ppt), the amphipods exhibit a respiratory and excretory response to hyper- and hypoosmotic stress; however, a rather constant O:N atomic ratio (around 15) was obtained at the experimental salinities, indicating protein/lipids as metabolic substrate. Both rates of oxygen consumption and ammonia excretion increased with an increasing osmotic difference (0 to 650 mOsm) between the haemolymph and the environmental medium, indicating higher energy requirements for osmotic and ionic regulation at low salinities. In amphipods abruptly transferred from 35 to 5 ppt, a minor decrease of the haemolymph sodium concentrations together with an increased ammonia excretion output indicate a counter-ion regulation of NH ₄ ⁺ and Na⁺ during hyposmotic stress.     

Light‐induced changes in rabbit pineal gland n‐acetyltransferase activity

N‐acetyltransferase (NAT) activity was determined in different ages of New Zealand White rabbit pineal gland using 2‐aminofluorene and p‐aminobenzoic acid as substrates and it was assayed by high pressure liquid chromatography. Rabbits of different ages were either sacrificed during the light phase, exposed to darkness or light for 1 min during the dark phase of the lighting cycle, returned to their cages in darkness for 30 min and then sacrificed. Pineal gland NAT activity in animal nocturnally exposed to 1 min of light was inhibited in animals 1 ‐day‐old of age. Nocturnal light exposure did not inhibit enzyme activity in 1‐day‐old rabbit, even though these animal displayed clear light : dark differences in pineal gland NAT activity. Nocturnal light exposure also did not inhibit night time levels of NAT activity in 1‐day‐old animals who had been bilaterally enucleated. The result suggested that this effect is retinally mediated. Pre‐treatment of 1‐day‐old and 60‐day‐old animals with the isoproterenol (beta‐noradrenoreceptor agonist drug), prevented the nocturnal light‐induced inhibition of NAT activity. The different sensitivity of 60‐day‐old and 1‐day‐old animals to different illuminances or durations of nocturnal light exposure, was that the duration or intensity of light exposure was enable to inhibit nocturnal NAT activity. The NAT activity was at least 3.2‐ to 4.6‐fold greater in 1‐day‐old rabbits compared to 60‐day‐old rabbits. Kinetic constants for arylamine NAT activity in pineal gland from rabbits were determined. K_m and F_max values for 2‐aminofluorene were 2.6‐fold higher for light exposure than for no light exposure rabbits. This is the first demonstration of the retina‐pineal gland pathway appears light‐induced changes in pineal glands of animals in 1‐day‐old of ages or older; but this pathway does not function in 60‐day‐old rabbits like manner in 1‐day‐old rabbits.     

Photoinhibition and recovery of the kelp Laminaria saccharina at optimal and superoptimal temperatures

Effects of high irradiance on photosynthetic characteristics were examined in sporophytes of the kelp Laminaria saccharina Lamour. from 1992 to 1994. Exposure to high irradiance (700 mol photons m^-2s^-1) for 1 h at optimal temperature (12°C) caused a 40 to 60% decline in photosynthetic efficiency (alpha), quantum yield, and the ratio of variable to maximum chlorophyll fluorescence (F_v/F_m), an indicator of Photosystem II efficiency. Although the photoinhibition effects were partly attributable to protective mechanisms, a concurrent increase in minimal fluorescence (F_o) indicated damage to Photosystem II reaction centers. The magnitude of photoinhibition was proportional to irradiance and duration; however, F_v/F_m was significantly reduced after exposure to irradiances as low as 40 to 50 mol photons m^-2s^-1 for 1 h, or to 700 mol photons m^-2s^-1 for only 5 min. In contrast, photosynthetic capacity (P_max) was affected only at much higher irradiance. Superoptimal temperatures up to 24°C did not exacerbate high-light effects. At 25°C, however, alpha and P_max were more susceptible to photoinhibition than at lower temperatures. Recovery from photoinhibition was examined by following F_v/F_m and F_o for 24 h after exposure to high light. Recovery of F_v/F_m was fastest during the first 1 to 3 h, and slowed or ceased after 6 to 12 h, while recovery of F_o was relatively constant over 12 h. Dithiothreitol, which blocks formation of energy-dissipating xanthophylls, reduced both the initial rate and extent of recovery. Chloramphenicol, which blocks chloroplast-encoded protein synthesis, had little effect on initial rates of recovery, but stopped recovery after 3 h. Thus, L. saccharina appears to rely on the xanthophyll cycle to protect the photosynthetic apparatus, and reversal of this protective mechanism causes the rapid initial recovery in F_v/F_m. Longterm recovery depends on repair of damaged reaction centers. Both the rate and extent of recovery were temperature-dependent. The initial rate was higher at 18 to 22°C than at 12°C, but the extent of recovery over 24 h declined with increasing temperature. High temperatures, therefore, appear to enhance protective mechanisms, but disrupt repair processes. L. saccharina from Long Island Sound, an ecotype adapted to low light and high temperature, showed slightly but consistently greater effects of photoinhibition than plants from the Atlantic coast of Maine, but exhibited faster recovery at superoptimal temperatures.     

Effects of UV-B radiation on the gills of Catla catla during early development

Scanning electron microscopic (SEM) study of gills of Catla catla catla (17-day-old) exposed to UV-B radiation (145?µW?cm^?2 at the water surface) for three different exposure times: 5, 10 and 15?min was conducted. Fish without UV-B exposure served as control. UV-B radiation damaged both gill filaments and lamellae. The intensity of damage was minimal in 5?min exposed fish, followed by 10?min exposed fish and maximal in 15?min. The gill epithelium was severely damaged in 15?min irradiated fish compared to control. Pavement cells (PVCs) were damaged and the numbers of microridges within PVCs decreased. The deep boundary of PVC was not clear. In some area of gill epithelium, PVCs were destroyed and mitochondrion-rich cells (MRCs) were exposed. The 5?min exposure reduced the number of microridges in the PVCs, but the boundary of PVCs was still visible. MRCs in the gill epithelium were not exposed in 5?min exposed fish. The damage to PVCs and subsequent exposure of MRCs in UV-B irradiated fish may hamper respiratory functions and disturb osmoregulation in catla.     

Salinity stress and hydrogen peroxide regulation of antioxidant defense system in Ulva fasciata

The regulation of antioxidant defense system in macroalgae exposed to salinity stress was examined in Ulva fasciata Delile. As compared to the 30‰ control, a long-term (4 days) exposure to hyposaline (5, 15‰) and hypersaline (60, 90, 120, 150‰) conditions inhibited growth rate and TTC reduction ability. A decrease in maximum quantum efficiency (F _v/F _m ratio) and the maintenance of superoxide dismutase activity under salinity stress indicate the potential generation of reactive oxygen species in chloroplasts. An exposure to 15, 60, and 90‰ decreased seawater H₂O₂ contents but increased thallus H₂O₂ contents that are positively correlated with TBARS and peroxide contents. Alleviation of oxidative damage and H₂O₂ accumulation at 15 and 90‰ by a H₂O₂ scavenger, dimethylthiourea, suggests that oxidative damage occurring under moderate hyposaline and hypersaline conditions is ascribed to accumulated H₂O₂. Increased glutathione reductase activity and glutathione content and decreased ascorbate content are responsible for accumulated H₂O₂ at 15, 60, and 90‰, while ascorbate peroxidase activity increased only at salinity ≥ 90‰. Catalase and peroxidase activities also increased at 60 and 90‰ for H₂O₂ removal, but only catalase showed activity increase at 15‰. For the regeneration of ascorbate, the activities of both dehydroascorbate reductase and monodehydroascorbate reductase were increased at 5 and 15‰ while only monodehydroascorbate reductase activity increased at 60 and 90‰. It is hypothesized that the availability of antioxidants and the activities of antioxidant enzymes are increased in U. fasciata to cope with the oxidative stress occurring in hyposaline and hypersaline conditions.     

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.     

Uptake and elimination kinetics of triphenyltin hydroxide by two fish species

The uptake from water and the elimination of carbon‐14 radiolabelled triphenyltin hydroxide ([¹⁴C]TPTH) was studied in two fish species: guppy (Poecilia reticulata) and rainbow trout larvae (Salmo gairdneri). During all the experiments no steady state in fish was found. TPTH was rapidly taken up, while elimination was very slow. Uptake and elimination rate constants (k ₁ and k ₂, respectively), and a bioconcentration factor were estimated, assuming first order kinetics.

During eight days of exposure of guppy to TPTH an uptake rate constant k ₁ of 70±7L/kg.d and an elimination rate constant k ₂ of 0.005 ±0.029d ^‐1 was found. This resulted in a biconcentration factor of at least 2.1 × 10³L/kg (wet weight). Comparable results were obtained during a 30 days exposure experiment with guppy: k ₁ was 41±2L/kg.d, k ₂ 0.014 + 0.002d^‐1, and the bioconcentration factor was estimated to be 2.9 × 10³ L/kg (wet weight).

Four days exposure of rainbow trout larvae resulted in a it, of 22+ 2 L/kg.d, and a k ₂ of 0.031 ±0.007d^‐1. Using these k, and k ₂ values it was estimated that the biconcentration factor exceeds 650 L/kg (wet weight).