A phycocyanin probe as a tool for monitoring cyanobacteria in freshwater bodies

In many countries, the presence of cyanobacteria in freshwater bodies used for both drinking water and recreational purposes is under increasing public health attention. Water managers are considering how to implement monitoring that leads to a minimization of the risks incurred by the users of potentially contaminated sites. To address this question, this study involved assessing the performance of a submersible probe for measuring phycocyanin-specific fluorescence as a function of cyanobacterial biomass, with the aim of applying it as a tool for surveillance management. Its advantages and limits compared to more traditional analyses are discussed. The monitoring of cyanobacteria in the water bodies of western France was carried out using a minifluorimeter specific to the fluorescence of phycocyanin, a pigment specific to cyanobacteria. The results are compared with the analyses recommended by the World Health Organisation (chlorophyll a and cell counting). This study based on nearly 800 samples shows a significant correlation between the phycocyanin content and the cyanobacterial biomass, expressed as the number of cells per mL (R2 = 0.73). This submersible probe is simple and rapid to use, making it possible to take into account horizontal and vertical heterogeneities in the proliferation growth. In this way, we are able to detect at an early stage the conditions that could potentially lead to a risk, in order to start sampling. Due to its sensitivity, this tool proves suitable for monitoring aimed at reducing the risks incurred by the users of contaminated sites and launching preventative actions. The use of the phycocyanin probe provides an effective tool to complement traditional analyses of cyanobacterial presence. It is suggested that a surveillance protocol based on phycocyanin concentration can significantly improved the accuracy of the extent of cyanobacterial bloom development in the light of spatial and temporal variabilities associated with these occurrences.     

Determination of dominant biogeochemical processes in a contaminated aquifer-wetland system using multivariate statistical analysis

Determining the processes governing aqueous biogeochemistry in a wetland hydrologically linked to an underlying contaminated aquifer is challenging due to the complex exchange between the systems and their distinct responses to changes in precipitation, recharge, and biological activities. To evaluate temporal and spatial processes in the wetland-aquifer system, water samples were collected using cm-scale multi-chambered passive diffusion samplers (peepers) to span the wetland-aquifer interface over a period of 3 yr. Samples were analyzed for major cations and anions, methane, and a suite of organic acids resulting in a large dataset of over 8000 points, which was evaluated using multivariate statistics. Principal component analysis (PCA) was chosen with the purpose of exploring the sources of variation in the dataset to expose related variables and provide insight into the biogeochemical processes that control the water chemistry of the system. Factor scores computed from PCA were mapped by date and depth. Patterns observed suggest that (i) fermentation is the process controlling the greatest variability in the dataset and it peaks in May; (ii) iron and sulfate reduction were the dominant terminal electron-accepting processes in the system and were associated with fermentation but had more complex seasonal variability than fermentation; (iii) methanogenesis was also important and associated with bacterial utilization of minerals as a source of electron acceptors (e.g., barite BaSO(4)); and (iv) seasonal hydrological patterns (wet and dry periods) control the availability of electron acceptors through the reoxidation of reduced iron-sulfur species enhancing iron and sulfate reduction.     

Dynamics of atmospheric nitrogen deposition in a temperate calcareous forest soil

In temperate forest ecosystems, soil acts as a major sink for atmospheric N deposition. A (15)N labeling experiment in a hardwood forest on calcareous fluvisol was performed to study the processes involved. Low amounts of ammonium ((15)NH(4)(+)) or nitrate ((15)NO(3)(-)) were added to small plots. Soil samples were taken after periods ranging from 1 h to 1 yr. After 1 d, the litter layer retained approximately 28% of the (15)NH(4)(+) tracer and 19% of (15)NO(3)(-). The major fraction of deposited N went through the litter layer to reach the soil within the first hours following the tracer application. During the first day, a decrease in extractable (15)N in the soil was observed ((15)NH(4)(+): 50 to 5%; (15)NO(3)(-): 60 to 12%). During the same time, the amount of microbial (15)N remained almost constant and the (15)N immobilized in the soil (i.e., total (15)N recovered in the bulk soil minus extractable (15)N minus microbial (15)N) also decreased. Such results can therefore be understood as a net loss of (15)N from the soil. Such N loss is probably explained by NO(3)(-) leaching, which is enhanced by the well-developed soil structure. We presume that the N immobilization mainly occurs as an incorporation of deposited N into the soil organic matter. One year after the (15)N addition, recovery rates were similar and approximately three-quarters of the deposited N was recovered in the soil. We conclude that the processes relevant for the fate of atmospherically deposited N take place rapidly and that N recycling within the microbes-plants-soil organic matter (SOM) system prevents further losses in the long term.     

Probabilistic intake assessment of multiple compounds as a tool to quantify the nutritional-toxicological conflict related to seafood consumption

Seafood represents a natural source of valuable nutrients (e.g., long chain omega-3 poly-unsaturated fatty acids (LC n-3 PUFAs), vitamin D, and iodine), but the favourable health perception is troubled by the presence of contaminants (e.g., PCBs, dioxin-like compounds, and (methyl) mercury (MeHg)). This dualism forms the basis for an important public health conflict. The objective of this study was to calculate and evaluate the simultaneous intake of multiple beneficial and harmful compounds (LC n-3 PUFAs, vitamin D, iodine, (Me)Hg, PCBs, and dioxin-like compounds) via seafood consumption in Belgium. A methodology and a software module were developed for executing probabilistic assessments of the simultaneous intake of multiple compounds leading to better insight in the problematic nature of food items involving both health benefits and risks. The simulations concerning seafood consumption in Belgium predicted that, as far as only seafood consumption is concerned, the considered populations do not reach a sufficiently high intake for the three nutrients considered (LC n-3 PUFAs, vitamin D, and iodine). Regarding the contaminants, (Me)Hg contamination of seafood on the Belgian market does not seem to be an issue of major toxicological concern. In contrast, for dioxin-like compounds the tolerable daily intake is reached by people with high seafood consumption.     

Evaluation of the exposure methodology for risk-benefit assessment of seafood consumption

This study evaluated the methodology of using existing, international data to calculate the intake of nutrients and contaminants via seafood consumption on a national level. This was done by combining French seafood consumption data with two different datasets. The results showed that a significant correlation was found between the results obtained by using different data sets and that a high percentage of the population was classified in the same category (above or below the intake reference values). As such, the general conclusion that can be drawn from the results: 'whether a part of the population does exceed the toxicological reference values for certain contaminants and whether a part of the population does not reach the recommended intake for certain nutrients' remains the same for the two approaches. Therefore, it can be concluded that the existing, international probability distributions are useful when aiming to perform a cost-effective risk benefit evaluation of seafood consumption on national or regional level. When this evaluation does indicate that more detailed results are needed because a certain health risk exists, it can be decided that it is worth to spend more time and money for the analyses of national samples in order to collect more detailed results.     

Long-term fate of exogenous metals in a sandy Luvisol subjected to intensive irrigation with raw wastewater

From 1899 to 2002, sandy Luvisol in the Paris region has been intensively irrigated with raw wastewater, resulting in major soil pollution by metallic trace elements (MTE). To identify the soil phases implicated in retaining these metals, sequential extractions were performed on a solum irrigated with untreated wastewater and another reference solum. The endogenous and exogenous fractions of MTE in the contaminated soil were discriminated using correlations between MTE and major elements defined from unpolluted soils of the area. In the contaminated soil no exogenous lead and chromium are present below the surface horizon, whereas exogenous zinc and copper are found down to the base of the solum. The endogenous MTE are mainly found in the residual fraction. Exogenous MTE appear to be associated with organic matter in the surface horizon, and exogenous zinc seems to be readsorbed on iron and manganese oxyhydroxides in the underlying horizons.     

Detection of trisomy 21 by quantitative fluorescent–polymerase chain reaction in uncultured amniocytes

Prenatal diagnosis of fetal trisomy 21 is usually performed by cytogenetic analysis. This requires lengthy laboratory procedures, high costs and is unsuitable for large-scale screening of pregnant women. Today, trisomy 21 can be rapidly diagnosed within 24 h by molecular analysis of uncultured fetal cells using the semi-quantification of fluorescent PCR products from short tandem repeat (STR) polymorphic markers. The aim of our study was to test a chromosome quantification method on the basis of the analysis of fluorescent PCR products derived from non-polymorphic target genes. Co-amplification of a portion of DSCR1 (Down syndrome Critical Region 1) and the reference gene, CFTR (cystic fibrosis transmembrane regulator) enabled molecular detection of trisomy 21. Our method was successfully tested on a total of 154 amniotic fluids in a blind prospective study. Calculation of the DSCR1/CFTR ratio allowed us to distinguish between 152 normal amniotic fluids (mean ratio 0.99) and 2 amniotic fluids presenting a trisomy 21 status (DSCR1/CFTR ratio of 1.53 and 1.61, respectively). The results obtained by conventional cytogenetic analysis and our quantitative PCR method were concordant in every case. Our gene-based fluorescent PCR approach represents an alternative molecular method for rapid and reliable detection of trisomy 21, which can be helpful in the prenatal diagnosis of women at high risk of fetal trisomy 21. Copyright © 2003 John Wiley & Sons, Ltd.     

Natural formation and degradation of chloroacetic acids and volatile organochlorines in forest soil--challenges to understanding

- DOI: http:/dx.doi.org/10.1065/espr2005.06.262 Goal, Scope and Background The anthropogenic environmental emissions of chloroacetic acids and volatile organochlorines have been under scrutiny in recent years because the two compound groups are suspected to contribute to forest dieback and stratospheric ozone destruction, respectively. The two organochlorine groups are linked because the atmospheric photochemical oxidation of some volatile organochlorine compounds is one source of phytotoxic chloroacetic acids in the environment. Moreover, both groups are produced in higher amounts by natural chlorination of organic matter, e.g. by soil microorganisms, marine macroalgae and salt lake bacteria, and show similar metabolism pathways. Elucidating the origin and fate of these organohalogens is necessary to implement actions to counteract environmental problems caused by these compounds. Main Features While the anthropogenic sources of chloroacetic acids and volatile organochlorines are relatively well-known and within human control, knowledge of relevant natural processes is scarce and fragmented. This article reviews current knowledge on natural formation and degradation processes of chloroacetic acids and volatile organochlorines in forest soils, with particular emphasis on processes in the rhizosphere, and discusses future studies necessary to understand the role of forest soils in the formation and degradation of these compounds. Results and Discussion Reviewing the present knowledge of the natural formation and degradation processes of chloroacetic acids and volatile organochlorines in forest soil has revealed gaps in knowledge regarding the actual mechanisms behind these processes. In particular, there remains insufficient quantification of reliable budgets and rates of formation and degradation of chloroacetic acids and volatile organochlorines in forest soil (both biotic and abiotic processes) to evaluate the strength of forest ecosystems regarding the emission and uptake of chloroacetic acids and volatile organochlorines, both on a regional scale and on a global scale. Conclusion It is concluded that the overall role of forest soil as a source and/or sink for chloroacetic acids and volatile organochlorines is still unclear; the available laboratory and field data reveal only bits of the puzzle. Detailed knowledge of the natural degradation and formation processes in forest soil is important to evaluate the strength of forest ecosystems for the emission and uptake of chloroacetic acids and volatile organochlorines, both on a regional scale and on a global scale. Recommendation and Perspective As the natural formation and degradation processes of chloroacetic acids and volatile organochlorines in forest soil can be influenced by human activities, evaluation of the extent of this influence will help to identify what future actions are needed to reduce human influences and thus prevent further damage to the environment and to human health caused by these compounds.     

Primary co-culture as a complementary approach to explore the diversity of bacterial associations in marine invertebrates: the example of <Emphasis Type="Italic">Nautilus macromphalus</Emphasis> (Cephalopoda: Nautiloidea)

The recent application of molecular tools to address associations between bacteria and marine invertebrates has provided access to an immense diversity of unidentified microbes resistant to cultivation. However, the role of bacteria as partners in animal physiology remains unclear and in most cases difficult to investigate in the absence of adequate condition of cell growth and proliferation. In this work, we studied the reservoir of microbes associated with the excretory organs of Nautilus macromphalus as a model. Using the bacterial 16S RNA gene as a marker, we compared three complementary approaches for bacterial detection: bacterial DNA extraction from N. macromphalus tissues (“molecular approach”), strain isolation to provide a bacterial culture collection (“microbiological approach”) and finally, maintenance of N. macromphalus excretory organ cells with associated bacteria (“cellular approach”). Our results stress the potential of the “cellular approach” as a promising new tool as it promotes the detection of as yet uncultured β-proteobacteria and spirochaetes associated with N. macromphalus, and serves as a foundation for future studies describing potential roles that these bacteria may play in Nautilus.     

Disposition and metabolic profiling of [14C]-decabromodiphenyl ether in pregnant Wistar rats

Fully brominated diphenyl ether, decabromodiphenyl ether (DBDE), is one of the most widely used brominated flame retardants worldwide. Little data is available about the metabolic fate of DBDE in animal models and nothing at all about the extent of foetal exposure. In this work, pregnant Wistar rats were force-fed with 99.8% pure [14C]-DBDE over 96 h at a late stage of gestation (days 16 to 19). More than 19% of the administered dose was recovered in tissues and carcasses, demonstrating efficient absorption of DBDE despite its high molecular weight and low solubility. The highest concentrations of DBDE residues were found in endocrine glands (adrenals, ovaries) and in the liver, with lower values recorded for fat. In all tissue extracts, most of the radioactivity was associated with unchanged DBDE. The use of high-grade purity [14C]-DBDE allowed quantification of several metabolites present both in maternal tissues and in foetuses. These biotransformation products accounted for 9-27% of the extractable radioactivity in tissues and 14% of that in foetuses. Three nona-BDEs and one octa-BDE were identified by LC-APPI/MS. The unequivocal characterisation of a hydroxylated octa-BDE isolated from liver was confirmed by NMR. In rat, the main metabolic pathways of DBDE are debromination and oxidation. DBDE, and very likely most of its metabolites, are able to cross the placental barrier in rat. Metabolic profiles, obtained in vivo for the first time, demonstrated the presence of DBDE and major biotransformation products in endocrine glands as well as in foetuses. The biological activity of these metabolites still needs to be assessed in order to better understand the potential toxicity of DBDE.