Microbial diversity and ecological networks as indicators of environmental quality

Evaluating the quality of ecosystems in terms of biological patrimony and functioning is of critical importance in the actual context of intensified human activities. Microbial diversity is commonly used as a bioindicator of ecosystems functioning. However, there is a lack of sensitivity of microbial diversity indicators in the case of moderate and chronic environmental degradation, such as atmospheric deposition of pollutants, agricultural practices, diffuse pollution by wastewater and climate change. As a consequence, there is a need for alternative bioindicators of soils and water quality. Here, we discuss the interest of adopting a more integrative approach based on biotic interaction networks beyond the simple diversity indicators. We review how the various biotic interactions can be integrated in the various microbial networks such as trophic, mutualistic and co-occurrence networks. Then we discuss the efficiency of microbial networks and associated metrics to detect changes in microbial communities. We conclude that the connectance, the number of links and the average degree of co-occurrence networks could vary from 10 to 50% in response to minor perturbations when microbial diversity parameters remain stable. Finally, we analyze studies that aimed at linking microbial networks and activity to evaluate the potential of such networks for providing simple and operational indicators of ecosystem quality and functioning.     

A half-century analysis of landscape dynamics in southern Québec,Canada

We studied landscape dynamics for three time periods (<1950, 1965, and 1997) along a gradient of agricultural intensity from highly intensive agriculture to forested areas in southern Québec. Air photos were analyzed to obtain long-term information on land cover (crop and habitat types) and linear habitats (hedgerows and riparian habitats) and landscape metrics were calculated to quantify changes in habitat configuration. Anthropogenic areas increased in all types of landscapes but mostly occurred in the highly disturbed cash crop dominated landscape. Perennial crops (pasture and hayfields) were largely converted into annual crops (corn and soybean) between 1965 and 1997. The coalescence of annual crop fields resulted in a more homogeneous agricultural landscape. Old fields and forest cover was consistently low and forest fragmentation remained stable through time in the intensive agriculture landscapes. However, forest cover increased and forest fragmentation receded in the forest-dominated landscapes following farm abandonment and the transition of old fields into forests. Tree-dominated hedgerows and riparian habitats increased in areas with intensive agriculture. Observed changes in land cover classes are related to proximate factors, such as surficial deposits and topography. Agriculture intensification occurred in areas highly suitable for agriculture whereas farm abandonment was observed in poor-quality agriculture terrains. Large-scale conversion of perennial crops into annual crops along with continued urbanization exerts strong pressures on residual natural habitats and their inhabiting wildlife. The afforestation process occurring in the more forested landscapes along with the addition of tree-dominated hedgerows and riparian habitats in the agriculture-dominated landscapes should improve landscape ecological value.     

Analysis of the Structure-Properties Relationships of Different Multiphase Systems Based on Plasticized Poly(Lactic Acid)

Poly(lactic acid) is one of the most promising biobased and biodegradable polymers for food packaging, an application which requires good mechanical and barrier properties. In order to improve the mechanical properties, in particular the flexibility, PLA plasticization is required. However, plasticization induces generally a decrease in the barrier properties. Acetyl tributyl citrate (ATBC) and poly(ethylene glycol) 300 (PEG), highly recommended as plasticizers for PLA, were added up to 17 wt% in P(D,L)LA. In the case of PEG, a phase separation was observed for plasticizer contents higher than 5 wt%. Contrary to PEG, the T_g decrease due to ATBC addition, modelled with Fox’s law, and the absence of phase separation, up to 17 wt% of plasticizer, confirm the miscibility of PLA and ATBC. Contents equal or higher than 13 wt% of ATBC yielded a substantial improvement of the elongation at break, becoming higher than 300%. The effect of PLA plasticization on the barrier properties was assessed by different molecules, with increasing interaction with the formulated material, such as helium, an inert gas, and oxygen and water vapour. In comparison to the neat sample, barrier properties against helium were maintained when PLA was plasticized with up to 17 wt% of ATBC. The oxygen permeability coefficient and the water vapour transmission rate doubled for mixtures with 17 wt% ATBC in PLA, but increased five-fold in the PEG plasticized samples. This result is most likely caused by increased solubility of oxygen and water in the PEG phase due to their mutual miscibility. To conclude, ATBC increases efficiently the elongation at break of PLA while maintaining the permeability coefficient of helium and keeping the barrier properties against oxygen and water vapour in the same order of magnitude.     

Detection of environmental clastogens and aneugens in human fibroblasts by cytokinesis-blocked micronucleus assay associated with immunofluorescent staining of CENP-A in micronuclei

The cytokinesis-blocked micronucleus (CBMN) assay, in combination with fluorescent in situ hybridization (FISH) of human pan-centromeric DNA probes, or with CREST antibodies that specifically stain kinetochore proteins, is widely used on several cell types. It distinguishes micronuclei containing one or several whole chromosomes, which are positively labeled (centromere positive micronucleus, C+MN, due to aneugenic effect), or acentric chromosome fragments, which are unlabeled due to the absence of centromere (centromere negative micronucleus, C−MN, due to clastogenic effect). However, the very slight level of the centromeric signals obtained with the FISH technique on primary human fibroblasts, a cell type commonly used in environmental genetic toxicology, leads to great difficulties in distinguishing C+MN and C−MN. Furthermore, the CREST technique may lead to inappropriate results particularly with regards to variations in antibody composition between patient sera. Our results show that the in vitro CBMN, in combination with immunofluorescence staining of CENP-A (centromere protein A), efficiently screens genotoxicants for their ability to induce clastogenic and/or aneugenic effects. We propose the in vitro CBMN assay in combination with immunofluorescence staining of CENP-A as a suitable tool in environmental genotoxicity testing of primary human fibroblasts.     

Natural radioactivity in winter wheat from organic and conventional agricultural systems

The distribution of natural radionuclides was studied in winter wheat plants collected from three sites in Belgium during 2004-2007. Activity concentrations of ⁴⁰K, ²²⁶Ra, ²²⁸Ra and ²²⁸Th in organically and conventionally grown wheat, and in the corresponding soil samples, were determined using ultra low-level gamma-ray spectrometry. The observed soil-to-wheat concentration ratios were calculated for the different parts of the wheat plant (root, stem and grain) in the two agricultural systems (organic and conventional). There were large variations in radionuclide activity concentrations between the sites and fields, but no significant difference between conventionally and organically grown wheat plants was observed.     

Kinetic characterizing of soil trace metal availability using Soil/EDTA/Chelex mixture

The kinetic aspects are not usually tackled when mimicking the soil trace metal mobilization. In this work, a simple procedure is developed for measuring the kinetics of Pb, Cu and Cd transfer from the soil solid phase towards a resin sink. A ternary system of Soil/EDTA/Chelex was employed for mimicking the metal transfer from two agricultural soil samples into the Chelex. Two different kinetic regimes (P₁ and P₂) were observed. The kinetic profile of Pb was distinctly different from those of Cd and Cu. Basing on kinetic principles, two kinetic models were proposed for estimating the apparent rate constants of leaching and removal processes in, respectively, two binary mixtures of soil/EDTA and EDTA extracts/Chelex. Contrary to Pb, solid phase pools of Cd and Cu exchanged with the solution on short time scales. The kinetic rate of desorption occurred in following order: Pb < Cu ? Cd. Comparing the kinetics of binary systems, the desorption process was hypothesized to control the metal transfer. The parameters related to the desorption process were extended to the kinetic regimes of Soil/EDTA/Chelex and revealed that the soil to Chelex transfer of Pb is under kinetics control whereas the bulk concentration (affected by the size of particulate reservoir) is likely to control the Cd transfer. Cu presents an intermediate case.     

Ecotoxicological risk assessment linked to the discharge by hospitals of bio-accumulative pharmaceuticals into aquatic media: The case of mitotane

The release of hospital wastewater into the urban sewer networks contributes to the general contamination of aquatic media by pharmaceutical residues. These residues include bio-accumulative pharmaceuticals that lead to increased risk for ecosystems because they can concentrate in organisms and food chains, and therefore reach toxic levels. In order to assess the ecotoxicological risks linked to this particular category of residues, we have developed a specific method, by combining a theoretical calculation of pollutant concentrations in organisms to estimate Body Residue (BR), and ecotoxicity biomarkers in fish cell lines, enabling the calculation of a Critical Body Residue (CBR). This method finally results in the calculation of a specific risk quotient (Qb = BR/CBR), characterizing the risk linked to this type of pollutant. This method was applied to mitotane, a bio-accumulative pharmaceutical typically found in hospital wastewater, in the framework of an exposure scenario corresponding to the discharge of all the hospital wastewaters into the Rhone River which flows through the city of Lyon, France. This approach leads to risk quotients (Qb and Qbg) much higher than those found with the classical approach, i.e. Q = PEC/PNEC (Predictive Environmental Concentration/Predictive Non Effect Concentration) = 0.0006. This difference in the appreciation of risk is important when using cytotoxicity as the criterion for measuring the toxicity of mitotane (Qb = 0.056) and it is even greater when the criterion used is genotoxicity (Qbg = 6.8). This study must be now consolidated by taking the biomagnification of the pharmaceuticals into consideration.     

Organochlorinated pesticides, PCBs, dioxins, and PBDEs in grey mullets (Liza ramada) and allis shads (Alosa alosa) from the Vilaine estuary (France)

This study aimed to compare the contamination levels of various organohalogenated compounds in two migratory fish species in the Vilaine River in western France. Organochlorinated pesticides, polychlorinated biphenyls (PCBs), dioxins (polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/Fs)), and polybrominated diphenyl ethers (PBDEs) were analyzed in two diadromous species from the Vilaine estuary, the grey mullet (Liza ramada)—an amphihaline species, and the allis shad (Alosa alosa)—an anadromous species. Fish were collected in spring 2004 and spring 2005, upstream and downstream of the Arzal Dam. PCB contamination varied from 27 to 200 ng?g^?1 dry weight (d.w.). PCDDs/Fs, expressed in toxicity equivalent quantity (TEQ) varied from 0.4 to 2.8 pg?g^?1 d.w. Dioxins and dioxin-like PCBs expressed in total TEQ varied from 1.4 to 18.8 pg?g^?1 d.w. PBDE47 was present at around 2–10 ng?g^?1 d.w. and concentrations of the insecticide dichlorodiphenyltrichloroethane breakdown product p,p′-dichlorodiphenylchloroethylene varied from 1 to 14 ng?g^?1 d.w. For both species, specimens collected upstream were more contaminated. The grey mullet specimens were less contaminated than the allis shad when taken downstream of the dam but were more contaminated upstream. The allis shads presented intermediate contaminant concentrations with a less pronounced difference between upstream and downstream specimens. However, it is thought that shads do not feed when they spawn in the upstream parts of rivers, which should modify the contaminant concentrations. However, measurements in upstream shad samples show an unexpected increase of the contamination, which remains unexplained.     

The ‘Storm of the Century’ (December 1999) and the accidental escape of Siberian sturgeons (Acipenser baerii) into the Gironde estuary (Southwest France)

GOAL, SCOPE AND BACKGROUND: The Gironde estuary is considered as a reference ecosystem with a large fish assemblage (75 species) and the presence of eleven diadromous species. However, geochemical studies and biomonitoring using oysters have shown high metal pollution levels. During the December 1999 hurricane, more than 5,000 young Siberian sturgeons (Acipenser baerii) accidentally escaped from a fish farm into the estuary, where this species had not been present until then. METHODS: In 2002/2003, thirteen sturgeons were collected from the estuary and twelve were obtained from the fish farm, from the same initial batch. Analysis of five metals (cadmium, zinc, copper, lead, mercury) measured in the gills, muscle, liver and kidneys, revealed significantly higher levels of cadmium, lead and mercury in the sturgeon collected from the estuary compared with those from the fish farm. RESULTS AND DISCUSSION: Metal analysis of the stomach contents of fish collected from the estuary and artificial food used in the fish farm indicated that dietary exposure might account for the differences observed. Nevertheless, metal concentrations in sturgeons that had spent about three years in the Gironde estuary may be considered low when compared with other species (mullets, eels) or with data in the literature regarding concentrations able to induce structural and functional perturbations in fish. As the introduced Siberian sturgeons live in the same place and feed on the same prey as the juveniles of the endangered A. sturio, our results indicate in the short-term that this strictly protected species does not appear to be much affected by the polymetallic pollution in the Gironde estuary. Marked differences in the concentrations of three metals (Cd, Pb, Hg) were established in four organs (gills, liver, skeletal muscle, kidneys) of Siberian sturgeons between specimens that had accidentally escaped into the Gironde estuary (France) during the December 1999 hurricane and were collected in March/June 2003 and specimens from the same initial batch at the fish farm. RECOMMENDATIONS AND PERSPECTIVES: Based on these results, a programme will be set up, with sampling campaigns carried out in the estuary every two years, with systematic monitoring of age in order to check the origin of individuals, in conjunction with sampling of sturgeon from the fish farm. This programme will complement other studies on several biological species representative of the main trophic levels in the estuary, especially the plankton component, benthic macroinvertebrates and oysters.     

Responses of the European flounder (Platichthys flesus) to a mixture of PAHs and PCBs in experimental conditions

A multibiomarker approach was developed to evaluate the juvenile European flounder responses to a complex mixture of 9 polycyclic aromatic hydrocarbons (PAHs) and 12 polychlorinated biphenyls (PCBs). Exposure was performed through contaminated food pellets displaying: (1) PAH and PCB levels similar to those detected in the heavily polluted Seine estuary, respectively in sediments and in flatfish and (2) ten times these concentrations. Several biomarkers of the immune system (e.g., lysozyme concentration and gene expression of complement component C3 and TNF-receptor), DNA damage (e.g., Comet assay), energetic metabolism (e.g.,?activity of cytochrome C oxidase), detoxification process (e.g., cytochrome P450 1A1 expression level: CYP1A1; betaine homocysteine methyl transferase expression level: BHMT) were investigated after 14 and 29 days of contamination, followed by a 14-days recovery period. After 29 days of contamination, the detoxification activity (CYP1A1 expression level) was positively correlated with DNA damages; the increase of the BHMT expression level could also be related to the detoxification process. Furthermore, after the recovery period, some biomarkers were still upregulated (i.e., CYP1A1 and BHMT expression levels). The immune system was significantly modulated by the chemical stress at the two concentration levels, and the lysozyme appeared to be the most sensitive marker of the mixture impact.