Mercury Increases Light Susceptibility in the Green Alga Haematococcus lacustris

The influence of mercury ions (Hg²⁺, 10 μM) on photosynthesis was investigated in flagellates and aplanospores of Haematococcus lacustris. Hg²⁺ stress resulted in a fast decrease of chlorophyll fluorescence yield. This was initially caused by an increase in reversible non-photochemical quenching of chlorophyll fluorescence. During further exposure to Hg²⁺, an increasing contribution of pH independent non-photochemical quenching and a parallel rise in the content of the xanthophyll cycle pigment zeaxanthin was detected. An increase of the initial chlorophyll fluorescence as a final sign of Hg²⁺ induced adverse effects on photosynthesis supports our hypothesis that mercury ions predispose to non-reversible, “chronic” photoinhibition.     

Prolonged incubation with low concentrations of mercury alters energy transfer and chlorophyll (Chl) a protein complexes in Synechococcus 6301: changes in Chl a absorption and emission characteristics and loss of the F695 emission band

Synechococcus PCC 6301 cells grown in the presence of low sublethal levels of (about 2 m) mercury induced alterations in chlorophyll (Chl) a absorption without significant alterations in phycocyanin. Chl a fluorescence emission in Hg²⁺ -raised cells showed a large (about 18 nm) blue shift in the peak emission. No major spectral changes in phycobilisome (PBsome) emission characteristic were noticed, indicating major structural alterations in Chl-protein complexes by incubation with Hg²⁺ ions. Low temperature (77 K) emission spectra of cells grown in the presence of Hg²⁺ showed a loss of the characteristic Chl a emission band at 695 nm (F695), which is known to be linked to photosystem II photochemistry and to originate from the Chl a of core antenna polypeptide CP 47 of photosystem II. The SDS-PAGE polypeptide profile of thylakoids indicates a loss of a polypeptide(s) with a molecular mass between 40 and 60 k Da by Hg²⁺ incubation of cells. Our results suggest that prolonged incubation of Synechococcus 6301 cells with low concentrations of Hg²⁺ affects the Chl a spectral properties and the structure of Chl-protein complexes.     

Action of selected heavy metal ions on the photosystem 2 activity of the cyanobacteriumSpirulina platensis

Addition of different concentrations of heavy metal ions (Hg²⁺, Cu²⁺, Ni²⁺ and Pb²⁺) inhibited the photosystem 2 catalyzed electron transport activity (H₂O→p-benzo-quinone) of the cyanobacteriumSpirulina platensis. Hg²⁺ caused the inhibition in electron transport activity in very low concentrations compared to the other metal ions. Hg²⁺ at this low concentration specifically altered the spectral properties of phycocyanin of the phycobilisomes in the intact cells ofSpirulina, whereas other heavy metal ions were ineffective in this sense.     

Functional aspects of secondary carotenoids in Haematococcus lacustris (Girod) Rostafinski (Volvocales). V. Influences on photomovement

Secondary carotenoids are suspected to modulate photomovement in Haematococcus lacustris [Girod] Rostafinski (Volvocales). To investigate the influence of these extrachloroplastic ketocarotenoids on phototactic and photophobic responses in the flagellate stage of the green alga, flagellate suspensions differing in the content of secondary carotenoids were grown from green and red aplanospores. Photo-orientation of these flagellates induced by unilateral irradiation was investigated using a computer-aided system for microscopic image analysis. Results were hypothetically summarized as follows: (1) Diminution of precision of the positive phototaxis was found in red flagellates. This might be due to cellular shading of the blue-light-sensitive photoreceptor by secondary carotenoids. (2) Red flagellates exhibited an increase in the photophobic response. This finding is discussed in relation to an adaptive increase of the photoreceptor sensitivity, thought to be a result of the higher optical density of the corresponding cell suspension in the blue wavelength region.     

Mercury ions inhibit photosynthetic electron transport at multiple sites in the cyanobacteriumSynechococcus 6301

Inhibition of electron transport activities in the spheroplasts ofSynechococcus 6301 by HgCl² is dependent on the concentration of mercury ions. The inhibition of whole chain electron transport activity occurs at low concentration of Hg²⁺ (6 ΜM@#@). This inhibition occurs mostly due to interaction of Hg²⁺ on plastocyanin. At an elevated concentration (24 ΜM@#@), mercury induces inhibition chiefly in photosystem II catalyzed electron transport. At this concentration it also alters both the absorption and emission characteristics of the phycocyanin. The photosystem I catalyzed electron transport was inhibited by 50% only at high concentrations (36 ΜM@#@) of HgCl₂. However, electron transport catalyzed by photosystems I and II from reduced duroquinone to methylviologen which involves intersystem electron transport is extremely sensitive to mercury (low concentration 6–9 ΜM) like that of whole chain assay indicating that the observed inhibition in whole chain electron transport at low concentrations is mostly contributed by the damage involving other intersystem electron transport carrier(s) like plastocyanin. Thus mercury ions depending on the concentration affects the electron transport at multiple sites in the spheroplasts ofSynechococcus.     

Isolation of a mercuric chloride-tolerant bacterium and uptake of mercury by the bacterium

Summary A mercuric chloride-tolerant bacterium was isolated from activated sludge and the bacterium was identified as Pseudomonas oleovorans.The bacterium is tolerant up to 350 ppm Hg²⁺, 100 ppm Cd²⁺, 40 ppm cr⁶⁺, and 1000 ppm Cu²⁺. Observation by scanning electron microscope of cells growing with mercury shows a less rigid structure of the cell surface than in the case of controls, while observation by transmission electron microscope shows many electron-dense granules in the cytoplasm of cells growing with mercury. By chemical analysis, about 80% of mercury taken up by the cells was found in the cytoplasm, and about 20% in the cell envelope fraction, of the resting cells used. Mercury was found in the supernatant fluid, but not in the microsomal fraction.Further detailed experiments for mercury distribution in the cells show that it is concentrated in the nucleic acid fraction of the supernatant fluid. In the insoluble fraction, mercury was found mainly in the polyphosphate-polysaccharide, mucopeptide, and lipopolysaccharide-polysaccharide fractions. In the case of the growing cells used, about 20% of the mercury was found in the cytoplasm and 80% in the insoluble fraction.     

Effects of lanthanum and calcium on photoelectron transport activity and the related protein complexes in chloroplast of cucumber leaves

The effects of lanthanum and calcium ions on electron transport, dichlorephenol indophenol (DCIP) photoreduction, and oxygen evolution activities in chloroplast from cucumber (Cucumis satives L.) were determined. The lanthanum inhibited the whole electron chain-transport activity of chloroplast. DCIP photoreduction and oxygen evolution activities of the photosystem I (PSII) also decrease after treatment with La³⁺. But the diminished activities of PSII and chloroplast caused by La³⁺ could be reversed by Ca²⁺ and even became higher than the control level. The concentration analysis of related protein complexes to photoelectron transport in chloroplast included that La³⁺ induced the concentration of chlorophyll protein complexes increasing but caused some nonchlorophyll protein complexes to decompose partially. This increasing effect of La³⁺ on chlorophyll protein complexes results in the improvement of chlorophyll content, which will improve the absorption of photoelectron and energy transport in the process of photosynthesis.     

Presynaptic and postsynaptic effects of mercuric ions on guinea-pig ileum longitudinal muscle strip preparation

The toxic effect of mercuric ions on intestinal cholinergic neurotransmission was investigated in vitro. Hg²⁺ inhibited the evoked release and enhanced the resting release of ACh. Smooth muscle contraction was irreversibly inhibited by Hg²⁺ in a concentration-dependent manner, and Na₂EDTA did not antagonize this effect. We also investigated if Hg²⁺ enters the nerve terminal through Ca²⁺-channels, or Na⁺-channels, or both. The effects of mercuric ions obtained in our study were completely abolished by the combined administration of TTX and Co²⁺. It is suggested that the site of the action of mercuric ions is intracellular. We concluded that Hg²⁺ may interfere with cholinergic transmission by blocking [Ca²⁺]_o-dependent release of ACh and by enhancing [Ca²⁺]_o-independent resting release of ACh. The effect of Hg²⁺ was not only presynaptic since it also inhibited the effect of ACh on smooth muscle.     

Facilitated permeation of methyl viologen into chloroplasts in situ during electric pulse generation in excitable plant cell membranes

Generation of action potential (AP) in plasma membranes of characean algae has a strong impact on photoreactions occurring in chloroplasts. Under physiological conditions, AP suppresses electron transport in alkaline and acidic regions, although to a different extent; these changes are transient and reversible. In the presence of the artificial electron acceptor, methyl viologen (MV²⁺), AP-induced changes in electron transport in photosystem II become irreversible. Incubation of Chara corallina internodal cells with MV²⁺ has no effect on the chlorophyll P700 photooxidation kinetics in photosystem I reaction centers, suggesting that MV²⁺ is inaccessible for interactions with photosystem I, because its permeation into chloroplasts of a resting cell is hindered by membrane barriers. At the same time, AP generation in the presence of MV²⁺ is accompanied by irreversible modification of P700 photooxidation kinetics, as can be evidenced from differences in absorption changes at 810 and 870 nm (ΔA ₈₁₀ signals). These findings suggest that MV²⁺ permeation into chloroplasts in situ is facilitated during or after the AP generation. Similar to the ΔA ₈₁₀ signals, light-induced changes in membrane potential do not depend on the presence of MV²⁺ in the external medium until the first excitatory stimulus is applied. Electric photoresponses of the cell are irreversibly modified by AP generated in the presence of MV²⁺ at the expense of non-cyclic photosynthetic electron transport redirected to the MV²⁺ reduction. It is concluded that AP effects on chloroplast photosynthesis in situ are complex and involve permeability changes for MV²⁺ in membrane barriers of the “plasmalemma-chloroplast envelope” system.     

Biosorption of inorganic and methyl mercury by a biosorbent from Aspergillus niger

A biosorbent prepared by alkaline extraction of Aspergillus niger biomass was evaluated for its potential to remove mercury species – inorganic (Hg²⁺) and methyl mercury (CH₃Hg⁺) – from aqueous solutions. Batch experiments were carried out to determine the pH and time profile of sorption for both species in the pH range 2–7. The Hg²⁺ exhibited more rapid sorption and higher capacity than the CH₃Hg⁺. Further, removal of both mercury species from spiked ground water samples was efficient and not influenced by other ions. Sorption studies with esterified biosorbent indicated loss of binding of both mercury species (>80%), which was regained when the ester groups were removed by alkaline hydrolysis, suggesting the involvement of carboxyl groups in binding. Further, no interconversion of sorbed species occurred on the biomass. The biosorbent was reusable up to six cycles without serious loss of binding capacity. Our results suggest that the biosorbent from Aspergillus niger can be used for removal of mercury and methyl mercury ions from polluted aqueous effluents.     

Alterations in renal cellular glutathione metabolism after in vivo administration of a subtoxic dose of mercuric chloride

Renal cellular concentration of glutathione (GSH) increases after exposure to a subtoxic dose of inorganic mercury (Hg²⁺). In the present study, we tested the hypothesis that the increase in renal cellular concentration of GSH after exposure to a subtoxic dose of Hg²⁺ (0.5 μmol HgCl₂/kg body wt) is due to induction of GSH synthesis. Rats were treated in vivo with HgCl₂, and renal proximal tubular (PT) and distal tubular (DT) cells were isolated 24 hours later. PT cells were studied as the presumed target site for Hg²⁺, and DT cells were investigated as a nontarget cell population. γ-Glutamylcysteine synthetase activity increased after exposure to Hg²⁺ in PT cells when expressed on a per cell basis. Increases in activities of glutathione disulfide (GSSG) reductase, GSH peroxidase, and several enzymes involved in cellular energetics occurred after exposure to Hg²⁺. Many of these increases were observed in both PT and DT cells, indicating that the responses to Hg²⁺ were not restricted to the PT cells. These results are consistent with the hypothesis that in vivo exposure to a subtoxic dose of Hg²⁺ is also associated with induction of GSH synthesis and other key cellular enzymes. Early changes in GSH metabolism associated with exposure to Hg²⁺ appear to occur both in the primary target cell population and in more distal nephron sites. © 1996 John Wiley & Sons, Inc.     

A coumarin‐based fluorescent probe for Hg2+ and its application in living cells and zebrafish

Mercury (Hg) is a heavy metal with high toxicity and easy migration; it can be enriched through the food chain, and cause serious threats to the natural environment and human health. So, the development of a method that can be used to detect mercury ions (Hg²⁺) in the environment, in cells, and in organisms is very important. Here, a new 7‐hydroxycoumarin‐derived carbonothioate‐based probe ( CC‐Hg ) was designed and synthesized for detection of Hg²⁺. After addition of Hg²⁺, a large fluorescence enhancement was observed due to the formation of 7‐hydroxyl, which reinforced the intramolecular charge transfer process. The CC‐Hg probe had good water solubility and selectivity. Moreover, the probe was able to detect Hg²⁺ quantitatively over the concentration range 0–2 μM and with a detection limit of 7.9 nM. Importantly, we successfully applied the probe to detect Hg²⁺ in water samples, in living cells, and in zebrafish. The experimental results demonstrated its potential value in practical applications.     

Characterization of mercury(II)-induced inhibition of photochemistry in the reaction center of photosynthetic bacteria

Mercuric contamination of aqueous cultures results in impairment of viability of photosynthetic bacteria primarily by inhibition of the photochemistry of the reaction center (RC) protein. Isolated reaction centers (RCs) from Rhodobacter sphaeroides were exposed to Hg²⁺ ions up to saturation concentration (~?10³ [Hg²⁺]/[RC]) and the gradual time- and concentration-dependent loss of the photochemical activity was monitored. The vast majority of Hg²⁺ ions (about 500 [Hg²⁺]/[RC]) had low affinity for the RC [binding constant K_b?~?5 mM^?1] and only a few (~?1 [Hg²⁺]/[RC]) exhibited strong binding (K_b?~?50 μM^?1). Neither type of binding site had specific and harmful effects on the photochemistry of the RC. The primary charge separation was preserved even at saturation mercury(II) concentration, but essential further steps of stabilization and utilization were blocked already in the 5 < [Hg²⁺]/[RC]?<?50 range whose locations were revealed. (1) The proton gate at the cytoplasmic site had the highest affinity for Hg²⁺ binding (K_b?~?0.2 μM^?1) and blocked the proton uptake. (2) Reduced affinity (K_b?~?0.05 μM^?1) was measured for the mercury(II)-binding site close to the secondary quinone that resulted in inhibition of the interquinone electron transfer. (3) A similar affinity was observed close to the bacteriochlorophyll dimer causing slight energetic changes as evidenced by a?~?30 nm blue shift of the red absorption band, a 47 meV increase in the redox midpoint potential, and a?~?20 meV drop in free energy gap of the primary charge pair. The primary quinone was not perturbed upon mercury(II) treatment. Although the Hg²⁺ ions attack the RC in large number, the exertion of the harmful effect on photochemistry is not through mass action but rather a couple of well-defined targets. Bound to these sites, the Hg²⁺ ions can destroy H-bond structures, inhibit protein dynamics, block conformational gating mechanisms, and modify electrostatic profiles essential for electron and proton transfer.     

Wechselwirkungen zwischen Mikroorganismen und Metallionen

Microorganisms are able to interact with metal ions in aqueous solutions and to accumulate considerable amounts of them. The mechanism of the accumulation depends on the physiological state of the cells. In the case of resting cells the binding reaction takes place on the surface of the cell wall as a sorption process. The kinetic of the sorption processes in dependence of the physiological state of the cells, the concentration of Hg²⁺ and Cd²⁺-ions on the solution and the pH and temperature has been investigated. Beyond that possibilities for the desorption of the metals from the biomass have been tested.     

Carbon dots-modified paper-based chemiluminescence device for rapid determination of mercury (II) in cosmetics

Here, a simple and portable paper-based analytical device (PAD) based on the inherent capability of carbon quantum dots (CQDs) to serve as a great emitter for the bis(2,4,6-trichlorophenyl)oxalate (TCPO)–hydrogen peroxide (H₂O₂) chemiluminescence (CL) reaction is introduced for the detection of harmful mercury ions (Hg²⁺). The energy is transferred from the unstable reaction intermediate (1,2-dioxetanedione) to CQDs, as acceptors, and an intensive orange-red CL emission is generated at ~600 nm, which is equal to the fluorescence emission wavelength of CQDs. The analytical applicability of this system was examined for the determination of Hg²⁺. It was observed that Hg²⁺ could significantly quench the produced emission, which can be attributed to the formation of a stable and nonluminescent Hg²⁺–CQDs complex. Accordingly, a simple and rapid PAD was established for monitoring Hg²⁺, with a limit of detection of 0.04 μg ml⁻¹. No interfering effect on the signal was found from other examined cations, indicating the acceptable specificity of the method. The designed assay was appropriately utilized to detect Hg²⁺ ions in cosmetic samples with high efficiency. It was characterized by its low cost, ease of use, and was facile but accurate and high selective for the detection of Hg²⁺ ions. In addition, the portability of this probe makes it suitable for on-site screening purposes.     

Effects of Mercuric Chloride on Growth and Morphology of Selected Strains of Mercury-Resistant Bacteria

A survey of the comparative cytological effects of growth in the presence of mercury by a group of mercury-resistant bacterial cultures and a characterization of the process of bacterial adaptation to Hg²⁺ ion was accomplished. Mercury resistance was found to be dependent upon the ability to volatilize mercury from the medium and upon the amount of mercury accumulated by the cells. The results indicate that most cultures which adapt to growth in the presence of HgCl₂ exhibit extensive morphological abnormalities. Significant effects are delay in the onset of growth and cell division and numerous structural irregularities associated with cell wall and cytoplasmic membrane synthesis and function. A detailed analysis of the adaptation process and the resulting effects on morphology was performed on an Enterobacter sp. During the period preceding active multiplication, a selection for mercury-resistant mutants occurred. It was also demonstrated that growth commenced only at a specific threshold concentration of Hg²⁺.     

A highly selective colorimetric and long‐wavelength fluorescent probe for the detection of Hg2+

Currently, the fluorescent probe is an important method for detecting heavy metal ions, especially mercury ion (Hg²⁺), which is harmful to the health of humans and the environment due to its toxicity and extensive use. In this paper, we designed and synthesized a colorimetric and long‐wavelength fluorescent probe Hg‐P with high sensitivity and excellent selectivity, which could detect Hg²⁺ by the changes of visual color, fluorescence and absorption spectroscopy. With the addition of Hg²⁺ to probe Hg‐P solution, its color changed from yellow to pink, and showed a 171 nm red‐shifted absorption spectrum. Probe Hg‐P was used in real water and soil solution samples to detect Hg²⁺, and the result is satisfactory. Therefore, this new probe shows great value and application in detecting Hg²⁺ in the environment.     

EGTA, a calcium chelator, inhibits electron transport in photosystem II of spinach chloroplasts at two different sites

A fifteen minute incubation of spinach chloroplasts with the divalent Ca²⁺ chelator, EGTA, in concentrations 50–250 μM, inhibits electron transport through both photosystems. All photosystem II partial reactions, including indophenol, ferricyanide and the DCMU-insensitive silicomolybdate reduction are inhibited from 70–100%. The photosystem II donor reaction, diphenyl carbazide → indophenol, is also inhibited, indicating that the inhibition site comes after the Mn²⁺ site, and that the first Ca²⁺ effect noted (site II) is not on the water oxidation enzyme, as is commonly assumed, but between the Mn²⁺ site and plastoquinone A pool. The other photosystem II effect of EGTA (Ca²⁺ site I), occurs in the region between plastoquinone A and P700 in the electron transport chain of chloroplasts. About 50% inhibition of the reaction ascorbate + TMPD → methyl viologen is given by incubation with 200 μM EGTA for 15 min. Ca²⁺ site II activity can be restored with 20 mM CaCl₂. Ca²⁺ site I responds to Ca²⁺ and plastocyanin added jointly. More than 90% activity in the ascorbate + TMPD → methylviologen reaction can be restored. Various ways in which Ca²⁺ ions could affect chloroplast structure and function are discussed. Since EGTA is more likely to penetrate chloroplast membranes than EDTA, which is known to remove CF₁, the coupling factor, from chloroplast membranes, and since Mg²⁺ ions are ineffective in restoring activity, it is concluded that Ca²⁺ may function in the electron transport chain of chloroplasts in a hitherto unsuspected manner.     

Bioremoval capacity of three heavy metals by some microalgae species (Egyptian Isolates)

Three fresh water microalgal isolates [Phormidium ambiguum (Cyanobacterium), Pseudochlorococcum typicum and Scenedesmus quadricauda var quadrispina (Chlorophyta)] were tested for tolerance and removal of mercury (Hg²⁺), lead (Pb²⁺) and cadmium (Cd²⁺) in aqueous solutions as a single metal species at conc. 5–100 mg / L under controled laboratory conditions. The obtained results showed that Hg²⁺ was the most toxic of the three metal ions to the test algae even at low concentration (< 20 mg/L). While lower concentration of Pb²⁺ and Cd²⁺ (5–20 mg / L) enhanced the algal growth (chlorophyll a and protein), elevated concentrations (40–100 mg / L) were inhibitory to the growth. The results also revealed that Ph. ambiguum was the most sensitive alga to the three metal ions even at lower concentrations (5 and 10 mg / L) while P. typicum and S. quadricauda were more tolerant to high metal concentrations up to 100 mg / L. The bioremoval of heavy metal ions (Hg²⁺, Pb²⁺ and Cd²⁺) by P. typicum from aqueous solution showed that the highest percentage of metal bioremoval occurred in the first 30 min of contact recording 97% (Hg²⁺), 86% (Cd²⁺) and 70% (Pb²⁺). Transmission electron microscopy (TEM) was used to study the interaction between heavy metal ions and P. typicum cells. At ultrastructural level, an electron dense layers were detected on the algal cell surfaces when exposed to Cd, Hg and Pb. At the same time, dark spherical electron dense bodies were accumulated in the vacuoles of the algal cells exposed to Pb. Excessive accumulation of starch around the pyrenoids were recorded as well as deteriorations of the algal cell organelles exposed to the three metal ions.