THE EFFECT OF LABELING INTENSITY, ESTIMATED BY REAL-TIME CONFOCAL LASER SCANNING MICROSCOPY, ON FLOW CYTOMETRIC APPEARANCE AND IDENTIFICATION OF IMMUNOCHEMICALLY LABELED MARINE DINOFLAGELLATES

Two different fluorescein isothiocyanate (FITC) conjugates were used to analyze the effect of labeling intensity on the flow cytometric appearance of marine dinoflagellates labeled with antibodies that specifically recognized the outer cell wall. Location of the labeling was revealed by epifluorescence and real-time confocal laser scanning microscopy using an anti-rabbit IgG/FITC-conjugated secondary antiserum. Flow cytometric measurements showed that cells of Prorocentrum species labeled this way could not always be distinguished from unlabeled cells. The labeling intensity increased several times when a biotinylated anti-rabbit IgG secondary antiserum was used in combination with a streptavidin/FITC conjugate. Flow cytometry indicated that the labeling intensity had increased 50%, which resulted in an improved separation of clusters of labeled and unlabeled cells.</p

Chemical characteristics and limnology of Loskop Dam on the Olifants River (South Africa), in light of recent fish and crocodile mortalities

A declining crocodile population and fish mortalities attributed to pansteatitis, along with increasing blooms of Microcystis aeruginosa and Ceratium hirundinella, have led to serious concerns about water quality in Loskop Dam, on the Olifants River, South Africa. Major impacts include acid mine drainage and eutrophication associated with sewage effluent. However, the specific causes of pansteatitis remain elusive. In 2011 the water chemistry and limnology of Loskop Dam were studied to determine factors that may be influencing aquatic ecosystem health. Long-term monitoring data collected by the Department of Water Affairs were analysed for trends using a seasonal Mann-Kendall trend test, and were used to determine the trophic state of Loskop Dam using the Carlson index. Multiple sites were sampled which showed the reservoir was heterogeneous with regard to nutrient concentrations, algal biomass and dissolved metals. Specifically, the transitional zone was characterised by frequent algal blooms, resulting in fluctuating dissolved oxygen (range = 2.1–14.5 mg/ℓ) and pH (range = 7.35–10.59) levels. Using total phosphorus, Secchi depth, and chlorophyll-a concentrations, the trophic state of Loskop Dam was classified as meso- to eutrophic. Significant positive trends were observed in total (Tau = 0.422) and dissolved inorganic (Tau = 0.193) phosphorus.The reservoir showed a monomictic pattern of summer stratification (October to April) and holomictic winter circulation (June to July), with an increase in the depth and extent of anoxia in the hypolimnion when compared to previous research. Simultaneous elevated concentrations of manganese (>370 μg/ℓ) and iron in near-bottom water samples coincided with hypolimnetic anoxia. Aluminium concentrations exceeded the target water quality range (>10 μg/ℓ) during summer (December) in both surface and near-bottom water samples. We conclude that fish in Loskop Dam are periodically exposed to several physiological stressors including elevated ammonia, aluminium, iron and manganese and possibly hydrogen sulphide, as well as low dissolved oxygen. While these factors have never individually been linked to pansteatitis, their combined impacts have not been studied. To ensure the sustainability of Loskop Dam, catchment management plans must focus on reducing phosphorus inputs, and continue seeking treatment solutions for mine-water associated with abandoned and working coal mines.http://www.wrc.org.zaam2014mn201

Importance of organic matter lability for monomethylmercury production in sulfate-rich marine sediments

Sediment cores were collected from two shallow sites in the Venice Lagoon, Italy, in order to study the lability of organic matter and the methylation rate of inorganic Hg(II). Measurements were made of concentrations of total Hg and monomethylmercury (MMHg), Hg(II) methylation rates, concentrations of total organic carbon and total nitrogen in the sediments, and dissolved sulfate, sulfide, and alkalinity in sedimentary pore waters. A positive linear relationship was detected between the specific Hg(II) methylation rate constant and the fraction of total Hg comprised of MMHg (%MMHg/Hg), indicating that short-term Hg(II) methylation rate reflects a long-term accumulation of MMHg in sediment. In addition, the %MMHg/Hg and specific Hg(II) methylation rate constant in sediment increased with decreasing ratios of total organic carbon to total nitrogen (C/N), whereas concentrations of dissolved sulfate, sulfide, and alkalinity in pore water remained constant. This result suggests that the Hg(II) methylation rate was affected by lability of organic matter. In particular, surface sediments, which contained large fractions of fresh algal organic material (C/N = 5.8–7.8), showed higher Hg(II) methylation rates than did deeper sediments (C/N > 10). Our results indicate that the C/N ratio can be used as a proxy for the lability of organic matter that influences Hg(II) methylation rate in sulfate-rich marine sediments

Mercury concentration and monomethylmercury production in sediment: effect of dredged sediment reuse on bioconcentration for ragworms

The effect of dredged sediment reuse on the production and bioconcentration of monomethylmercury (MMHg) was investigated by examining sediments and ragworms found in dredge material banks and surrounding sites in the Venice Lagoon, Italy. Total Hg concentrations in the surface 20 cm of sediments were higher in the banks than in the surrounding sites, but MMHg concentrations were similar, which suggests reduced MMHg production in the banks. Monomethylmercury content in ragworms was significantly lower (p?<?0.05) in the banks than in the surrounding sites. In pore water, concentrations of both sulfate and Fe decreased with depth in the upper 20 cm of the dredge banks. In contrast, sulfate concentrations were constant with depth and large amounts of dissolved Fe occurred in the upper 20 cm of sediments of surrounding sites. Continuous sulfate reduction and possible precipitation of iron sulfide may decrease the production and bioconcentration of MMHg in the dredge material banks compared to the surrounding sites. Overall, the production of MMHg in sediments and its bioconcentration in benthic organisms were connected to the process of sediment diagenesis of organic matter through the control of pore water and sediment geochemistry involving iron and sulfur

Beryllium isotope geochemistry of hydrothermally altered sediments

cited By 16International audienceno abstrac

Sulfide and iron control on mercury speciation in anoxic estuarine sediment slurries

In order to understand the role of sulfate and Fe(III) reduction processes in the net production of monomethylmercury (MMHg), we amended anoxic sediment slurries collected from the Venice Lagoon, Italy, with inorganic Hg and either potential electron acceptors or metabolic byproducts of sulfate and Fe(III) reduction processes, gradually changing their concentrations. Addition of sulfide (final concentration: 0.2–6.3 mM) resulted in an exponential decrease in the sulfate reduction rate and MMHg concentration with increasing concentrations of sulfide. Based on this result, we argue that the concentration of dissolved sulfide is a critical factor controlling the sulfate reduction rate, and in turn, the net MMHg production at steady state. Addition of either Fe(II) (added concentration: 0–6.1 mM) or Fe(III) (added concentration: 0–3.5 mM) resulted in similar trends in the MMHg concentration, an increase with low levels of Fe additions and a subsequent decrease with high levels of Fe additions. The limited availability of dissolved Hg, associated with sulfide removal by precipitation of FeS, appears to inhibit the net MMHg production in high levels of Fe additions. There was a noticeable reduction in the net MMHg production in Fe(III)-amended slurries as compared to Fe(II)-amended ones, which could be caused by a decrease in the sulfate reduction rate. This agrees with the results of Hg methylation assays using the enrichment cultures of anaerobic bacteria: whereas the enrichment cultures of sulfate reducers showed significant production of MMHg (4.6% of amended Hg), those of Fe(III), Mn(IV), and nitrate reducers showed no production of MMHg. It appears that enhanced Fe(III)-reduction activities suppress the formation of MMHg in high sulfate estuarine sediments

Detection of Toxic Phytoplankton Species by Immunochemical Particle Analysis Based on Flow Cytometry

Particulate suspended matter in oceanic, coastal, and estuarine regions can be specifically marked immunochemically with a fluorescent probe using antisera recognizing antigens present on their surface. Of the particulate matter, phytoplankton is a major component. Toxic species that may form harmful blooms can be a direct threat to aquaculturing, tourism, sea-life and man. In order to detect such species in natural fixed phytoplankton populations, immunochemical tagging has been combined with flow cytometric evaluation. Microalgal cells can be labeled with a fluorescent probe (fluorescein isothiocyanate, FITC, is recommended). Labeled cells are counted using a flow cytometer. This method has proved to be applicable in a monitoring programme in the North Sea.

Biogeochemical factors affecting mercury methylation in sediments of the Venice Lagoon, Italy

Mercury methylation and sulfate reduction rates, total Hg, and monomethyl Hg in the sediments of the Venice Lagoon(Italy) were measured in June 2005 in order to identify the factors affecting the methylation of inorganic Hg. While the rates of Hg methylation and sulfate reduction were generally higher in the surface layers (0–2.5 cm), the correlation between Hg methylation and sulfate reduction rates was not significant when considering all depths and sites. This discrepancy is discussed considering two factors: the activity of sulfate-reducing bacteria and Hg solubility. The former factor is important in determining the Hg methylation rate in comparable geochemical conditions as evidenced by similar vertical profiles of Hg methylation and sulfate reduction rates in each sediment core. The latter factor was assessed by comparing the Hg methylation rate with the particle–water partition coefficient of Hg. The Hg methylation rates normalized to sulfate reduction rates showed a negative linear correlation with the logarithm of the particle–water partition coefficient of Hg, suggesting that the availability of dissolved Hg is a critical factor affecting Hg methylation. Solid FeS seems to play an important role in controlling the solubility of Hg in Venice Lagoon sediments, where sulfate and iron reductions are the dominant electron-accepting processes. Overall, the production of monomethyl Hg in the Venice Lagoon is controlled by a fine balance between microbial and geochemical processes with key factors being the microbial sulfate reduction rate and the availability of dissolved Hg

Mercury speciation in marine sediments under sulfate-limited conditions

Sediment profiles of total mercury (Hg) and monomethylmercury (MMHg) were determined from a 30-m drill hole located north of Venice, Italy. While the sediment profile of total Hg concentration was fairly constant between 1 and 10 m, that of the MMHg concentration showed an unexpected peak at a depth of 6 m. Due to the limited sulfate content (<1 mM) at the depth of 6 m, we hypothesized that the methylation of inorganic Hg(II) at this depth is associated with the syntrophic processes occurring between methanogens and sulfidogens. To test this hypothesis, anoxic sediment slurries were prepared using buried Venice Lagoon sediments amended with HgCl2, and we monitored MMHg concentration in sediment slurries over time under two geochemical conditions: high sulfate (1-16 mM) and limited sulfate concentrations (<100 µM). After day 52 and onward from the addition of inorganic Hg(II), the MMHg concentrations were higher in sulfate-limited slurries compared to high sulfate slurries, along with methane production in both slurries. On the basis of these results, we argue that active methylation of inorganic Hg(II) occurs under sulfate-limited conditions possibly by syntrophic processes occurring between methanogens and sulfidogens. The environmental significance of syntrophic Hg(II) methylation should be further studied