Oxidoreductases provide a more generic response to metallic stressors (Cu and Cd) than hydrolases in soil fungi: new ecotoxicological insights

The present study investigates the effect of metals on the secretion of enzymes from12 fungal strains maintained in liquid cultures. Hydrolases (acid phosphatase, β-glucosidase, β-galactosidase, and N-acetyl-β-glucosaminidase) and ligninolytic oxidoreductases (laccase, Mn, and lignin peroxidases) activities, as well as biomass production, were measured in culture fluids from fungi exposed to Cu or Cd. Our results showed that all fungi secreted most of the selected hydrolases and that about 50 % of them produced a partial oxidative system in the absence of metals. Then, exposure of fungi to metals led to the decrease in biomass production. At the enzymatic level, Cu and Cd modified the secretion profiles of soil fungi. The response of hydrolases to metals was contrasted and complex and depended on metal, enzyme, and fungal strain considered. By contrast, the metals always stimulated the activity of ligninolytic oxidoreductases in fungal strains. In some of them, oxidoreductases were specifically produced following metal exposure. Fungal oxidoreductases provide a more generic response than hydrolases, constituting thus a physiological basis for their use as biomarkers of metal exposure in soils.     

Simulation and evaluation of 2010 emission control scenarios in a Mediterranean area

At the onset of the 2010 statutory deadline for the respect of the European National Emission Ceiling directive, two questions arise. Will the engaged regulations for the respect of ozone air quality thresholds be fully efficient on the most polluted regions? How can we design the continuation of ozone control in those areas? This study is based on refined 3D modelling studies over a French Mediterranean region. It compares 2001 and 2003 situations with several prospective 2010 emission scenarios with, for the first time, the evaluation of local action plans. The degree of compliance with air quality regulation is investigated and the impact of emission control on the local potential for ozone formation is discussed. The results show that current efforts on emissions, although substantial and efficient, are not sufficient yet to abrogate all the ozone threshold exceedances. They also highlight the gap between regulatory and effective emission control, as well as the need for regional regulations to complete national efforts. Finally, the simulations indicate that large-scale emission control significantly helps reducing rural ozone (?20 ppbv) but affects much fewer (?2 to ?10 ppbv) the highest peaks. The continuation and the strengthening of ozone policies under their current form in such regions are considered.     

Managing Dive Tourism for the Sustainable Use of Coral Reefs: Validating Diver Perceptions of Attractive Site Features

It has been argued that strategies to manage natural areas important for tourism and recreation should integrate an understanding of tourist preferences for specific natural features. However, the accuracy of tourist recalled perceptions of environmental attributes, which are usually derived from post hoc surveys and used to establish management priorities, is currently unmeasured. We tested the validity of the relationship between tourist-stated preferences and actual condition of coral reefs around the Caribbean island of Bonaire. Using standardized questionnaires, we asked 200 divers to select their most and least favorite dive sites and the attributes that contributed to that selection. We also carried out ecological surveys at 76 of the 81 dives sites around the island to assess the actual conditions of the attributes indicated as important for site selection. Fish- and coral-related attributes were key features affecting dive enjoyment. In general, divers appeared to be able to perceive differences between sites in the true condition of biological attributes such as fish species richness, total number of fish schools, live coral cover, coral species richness, and reef structural complexity, although men and women divers differed in their ability to perceive/recall some of the attributes. Perceived differences in environmental attributes, such as surface conditions, underwater current, and the likelihood of encountering rare fish and sea turtles, were not empirically validated. The fact that divers perceive correctly differences in the condition of some of the key biological attributes that affect dive enjoyment reinforces the need to maintain overall reef condition at satisfactory levels. However, variation in accuracy of perceptions owing to demographic factors and attribute type suggests the need for caution when using public perceptions to develop environmental management strategies, particularly for coral reefs.     

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.     

Interaction of erythromycin ethylsuccinate and acetaminophen with protein fraction of extracellular polymeric substances (EPS) from various bacterial aggregates

Extracellular polymeric substances (EPS) are, along with microbial cells, the main components of the biological sludges used in wastewater treatment and natural biofilms. EPS play a major role in removing pollutants from water by means of sorption. The ability of soluble EPS (S-EPS) and bound EPS (B-EPS) derived from various bacterial aggregates (flocs, granules, biofilms) to bind at pH 7.0?±?0.1 to two pharmaceutical substances, acetaminophen (ACE) and erythromycin ethylsuccinate (ERY), has been investigated using the fluorescence quenching method. Two intense fluorescence peaks, A (Ex/Em range, 200–250/275–380 nm) and B (Ex/Em range, 260–320/275–360 nm), corresponding respectively to the aromatic protein region and soluble microbial by-product-like region, were identified in a three-dimensional excitation-emission matrix of EPS samples. The fluorescence peak, which corresponds to humic-like substances, was also identified though at low intensity. The ability of EPS to bind ACE was found to exceed that for ERY. The aromatic protein fraction of EPS displays a slightly higher affinity for drugs than that shown by the soluble microbial by-product-like fraction. The S-EPS and B-EPS present the same affinity for ACE and ERY. The effective quenching constants (log K) derived from the Stern–Volmer Equation equaled at peak A (with S-EPS): 3.7?±?0.2 to 4.0?±?0.1 for ACE and 2.1?±?0.3 to 2.7?±?0.1 for ERY. With B-EPS, these values were 3.9?±?0.1 to 4.0?±?0.1 for ACE and 2.0?±?0.2 to 2.6?±?0.1 for ERY. Our results suggest that the weaker EPS affinity for ERY than for ACE serves to partially explain why only about 50–80 % of ERY is removed from wastewater at the treatment plant. Moreover, this work demonstrates that EPS from natural river biofilms are able to bind drugs, which in turn may limit the mobility of drugs in natural waters.     

In vivo exposure of Mytilus edulis to living enteric bacteria: a threat for immune competency?

Mussels are widespread in coastal environments and experience various physical, chemical, and bacteriological conditions. Owing to the increase of coastal urbanization, mussels are now commonly exposed not only to indigenous bacteria, but also to enteric bacteria originating from pulsed and chronic sewage discharges into coastal environments. Due to its broad resilience to environmental variations, the blue mussel Mytilus edulis is commonly used as an indicator of environmental quality in bio-monitoring programs. However, since mussel immune system capabilities may be affected by the presence of exogenous fecal bacteria in coastal seawater subjected to sewage discharges, we aimed to determine the effect of in vivo bacterial challenges on mussels' immune competency by using two exogenous enteric bacterial strains, Escherichia coli and Enterococcus faecalis, and an indigenous bacterial strain Vibrio splendidus (as control). Bacterial strains were tested individually, by injection into the posterior adductor muscle at three different cell densities (10², 10³, and 10⁴ cells). Unlike classic in vitro experiments using higher bacterial concentrations, neither the enteric bacteria nor the indigenous strain induced significant increase or decrease of either cell-mediated (phagocytosis, reactive oxygen species, and NO _x production) or humoral components (prophenoloxidase-like, acid phosphatase, and l-leucine-aminopeptidase production) of the immune system. This study demonstrates that, at low concentrations, E. coli and E. faecalis do not represent an additional threat that could impair M. edulis immune competency and, as a consequence, its potential of survival in coastal areas subjected to sewage discharges.     

Methane oxidation in a crude oil contaminated aquifer: Delineation of aerobic reactions at the plume fringes

High resolution direct-push profiling over short vertical distances was used to investigate CH(4) attenuation in a petroleum contaminated aquifer near Bemidji, Minnesota. The contaminant plume was delineated using dissolved gases, redox sensitive components, major ions, carbon isotope ratios in CH(4) and CO(2), and the presence of methanotrophic bacteria. Sharp redox gradients were observed near the water table. Shifts in δ(13)C(CH4) from an average of -57.6‰ (±1.7‰) in the methanogenic zone to -39.6‰ (±8.7‰) at 105m downgradient, strongly suggest CH(4) attenuation through microbially mediated degradation. In the downgradient zone the aerobic/anaerobic transition is up to 0.5m below the water table suggesting that transport of O(2) across the water table is leading to aerobic degradation of CH(4) at this interface. Dissolved N(2) concentrations that exceeded those expected for water in equilibrium with the atmosphere indicated bubble entrapment followed by preferential stripping of O(2) through aerobic degradation of CH(4) or other hydrocarbons. Multivariate and cluster analysis were used to distinguish between areas of significant bubble entrapment and areas where other processes such as the infiltration of O(2) rich recharge water were important O(2) transport mechanisms.     

Charging for Nature: Marine Park Fees and Management from a User Perspective

User fees can contribute to the financial sustainability of marine protected areas (MPAs), yet they must be acceptable to users. We explore changes in the fee system and management of Bonaire National Marine Park (BNMP) from the perspective of users. Responses from 393 tourists indicated that 90% were satisfied with park conditions and considered current user fees reasonable. However, only 47% of divers and 40% of non-divers were prepared to pay more. Diver willingness-to-pay (WTP) appears to have decreased since 1991, but this difference could be due in part to methodological differences between studies. Although current fees are close to diver maximum stated WTP, revenues could potentially be increased by improving the current fee system in ways that users deem acceptable. This potential surplus highlights the value of understanding user perceptions toward MPA fees and management.     

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.     

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.