Successful analysis of gut contents in fungal-feeding oribatid mites by combining body-surface washing and PCR

The feeding habits of soil mesofauna have been a mystery for decades, and depending on the methods used, different degrees of feeding specialisation have been observed. A new way to study the almost unknown feeding habits of soil mesofauna e.g. oribatid mites is to use PCR-based techniques. When applying PCR on these small organisms, the low amount of ingested DNA can cause problems. Even more important is to certify that the amplified DNA does not originate from body-surface contamination. The aim of this study was to analyse if washing of the body surface combined with PCR can be a successful approach when identifying the food ingested by fungivorous mites. The method was developed in a laboratory system where we used the oribatid mite Archegozetes longisetosus as a model organism due to its relatively short life cycle and ease of laboratory culturing. The results demonstrated that surface contamination is a serious problem. Both washing and dissection was needed to remove surface contamination on such small organisms. To get a reliable result the samples also had to contain at least five pooled guts, but preferably ten. This is the first step towards a successful use of PCR-based methods to study natural feeding habits of species most likely contaminated on the body surface. When modified for field conditions, the results obtained by this method have a high potential to answer many questions about the animals feeding habits, and their functional role in the soil.     

Mate choice of an endogeic earthworm revealed by microsatellite markers

Endogeic earthworms are difficult study subjects due to the cryptic medium in which they live; thus, only the behaviour of epigeic and anecic earthworms has been studied before. We used microsatellite markers as a tool to elucidate the mate choice processes of Hormogaster elisae, an endogeic earthworm. It was shown to normally mate with two partners, preferably of the same size that are found in close proximity thereby eliminating the need for long-distance dispersion, which could explain the previously observed high genetic differentiation between populations. The genetic analyses of the sperm within each of its four spermathecae showed a uniform distribution with no signs of differential storage of sperm from different partners.     

Indirect host effect on ectomycorrhizal fungi: Leaf fall and litter quality explain changes in fungal communities on the roots of co-occurring Mediterranean oaks

Host trees can modify their soil abiotic conditions through their leaf fall quality which in turn may influence the ectomycorrhizal (ECM) fungal community composition. We investigated this indirect interaction using a causal modelling approach. We identified ECM fungi on the roots of two coexisting oak species growing in two forests in southern Spain - Quercus suber (evergreen) and Quercus canariensis (winter deciduous)-using a PCR-based molecular method. We also analysed the leaf fall, litter and soil sampled beneath the tree canopies to determine the concentrations of key nutrients. The total mycorrhizal pool was comprised of 69 operational taxonomic units (OTUs). Tomentella and Russula were the most species-rich, frequent and abundant genera. ECM fungi with epigeous and resupinate fruiting bodies were found in 60% and 34% of the identified mycorrhizas, respectively. The calcium content of litter, which was significantly higher beneath the winter-deciduous oak species due to differences in leaf fall quality, was the most important variable for explaining ECM species distribution. The evaluation of alternative causal models by the d-sep method revealed that only those considering indirect leaf fall-mediated host effects statistically matched the observed covariation patterns between host, environment (litter, topsoil, subsoil) and fungal community variables.     

First indications for the involvement of strigolactones on nodule formation in alfalfa (Medicago sativa)

Strigolactones have recently been suggested to be phytohormones that are present in all plants. Strigolactones are released by roots into the rhizosphere, stimulating the seed germination of parasitic plants such as Striga spp. and Orobanche spp. and play a crucial role in the interaction between plants and symbiotic arbuscular mycorrhizal fungi.By applying different concentrations of the synthetic strigolactone analogue GR24 to alfalfa (Medicago sativa) inoculated with Sinorhizobium meliloti we could show that in alfalfa nodulation is positively affected by the presence of the strigolactone analogue GR24. Moreover, we could show that this increased nodulation cannot be linked with a stimulatory effect of GR24 on the growth or the expression of nod genes of S. meliloti.Putative mechanisms operating in the plant in response to the addition of GR24 and leading to increased nodule formation by rhizobia are discussed.     

Emissions of CO2, CH4 and N2O from Southern European peatlands

Peatlands play an important role in emissions of the greenhouse gases CO₂, CH₄ and N₂O, which are produced during mineralization of the peat organic matter. To examine the influence of soil type (fen, bog soil) and environmental factors (temperature, groundwater level), emission of CO₂, CH₄ and N₂O and soil temperature and groundwater level were measured weekly or biweekly in loco over a one-year period at four sites located in Ljubljana Marsh, Slovenia using the static chamber technique. The study involved two fen and two bog soils differing in organic carbon and nitrogen content, pH, bulk density, water holding capacity and groundwater level. The lowest CO₂ fluxes occurred during the winter, fluxes of N₂O were highest during summer and early spring (February, March) and fluxes of CH₄ were highest during autumn. The temporal variation in CO₂ fluxes could be explained by seasonal temperature variations, whereas CH₄ and N₂O fluxes could be correlated to groundwater level and soil carbon content. The experimental sites were net sources of measured greenhouse gases except for the drained bog site, which was a net sink of CH₄. The mean fluxes of CO₂ ranged between 139 mg m⁻² h⁻¹ in the undrained bog and 206 mg m⁻² h⁻¹ in the drained fen; mean fluxes of CH₄ were between −0.04 mg m⁻² h⁻¹ in the drained bog and 0.05 mg m⁻² h⁻¹ in the drained fen; and mean fluxes of N₂O were between 0.43 mg m⁻² h⁻¹ in the drained fen and 1.03 mg m⁻² h⁻¹ in the drained bog. These results indicate that the examined peatlands emit similar amounts of CO₂ and CH₄ to peatlands in Central and Northern Europe and significantly higher amounts of N₂O.     

Heavy Metal and Nitrogen Monitoring Using Moss and Topsoil Samples in a Pyrenean Forest Catchment

This study was carried out in order to assess both the deposition of heavy metal and nitrogen in a mountain ecosystem with low levels of metal deposition and its possible interactions with factors such as lithology and topography. For this purpose, samples of Hypnum cupressiforme Hedw. and topsoils were collected in a forest catchment within Bertiz Natural Park, an International Cooperative Programme on Integrated Monitoring site. Trace metals levels in mosses can be considered low compared with values reported elsewhere in Europe, and the dust soil mineral particles seemed to be the main source of these values. Only Cd and Hg presented external inputs, probably with an anthropic origin, for mosses according to the enrichment factor values, whereas historical pollution-related deposition in soils was determined for Pb, Cu, and Ni, attending to their total/extractable ratio.     

Bioaccumulation of Copper by Zea mays: Impact on Root, Shoot and Leaf Growth

In the present study, the growth and the Cu²⁺accumulation by roots, shoots and leaves of Zea mays were examined using copper sulphate in the range of 10^?4 to 10^?2 M. Plants of Z. mays did not show inhibition of growth in the presence of 10^?4 to 10^?2 M Cu²⁺; however, it was observed growth effects on root when different Cu²⁺ solution concentrations were used. Only the seedlings exposed to 10^?2 M exhibited substantial root growth reduction, yielding only 56% of length with respect to the control. Seedlings exposed to 10^?4 M Cu²⁺ exhibited 16% and 42% growth increase in shoots and leaves, respectively, when compared with the controls. The seedlings treated with 10^?3 and 10^?2 M Cu²⁺ were inhibited in shoot and leaf growth. The fresh weights in roots, shoots and leaves significantly decreased at 10^?2 M Cu²⁺. The tolerance index, based on root length, was not significantly different for the three different treatments with copper. However, the total accumulation rate was very low at 10^?4 and 10^?3 M compared to 10^?2 Cu treatments. The capacity of copper accumulation by roots, shoots and leaves of Z. mays plants increased concomitant to the copper concentration, arriving to 382 times more in roots, 157 in shoots and only 16 in leaves, all compared to the controls. Cu could be accumulated by roots, shoots and leaves when the initial concentrations were 10^?3 and 10^?4 M. However, when it was 10^?2 M, the metal could not be accumulated by leaf and shoot levels; the roots could increase their copper accumulation capacity three times compared to the control. Z. mays has potential ability to accumulate Cu without being overly sensitive to Cu toxicity.     

Bioaccumulation and Effects of Heavy Metals in Crayfish: A Review

Metal pollution is a global problem which represents a growing threat to the environment. Because of bioaccumulation and negative effects of heavy metals, their bioavailability needs to be monitored. Many studies showed accumulation of metals in crayfish tissues as dose- and time-dependent without significant differences in tissue concentration levels comparing males and females. Muscles and exoskeleton were considered as specific for accumulation of mercury and nickel, respectively. Cadmium, zinc, copper, lead, and chromium accumulated mainly in hepatopancreas. By analyzing these specific tissues, it is possible to deduce the bioavailability and, by presumption, the level of environmental pollution by specific metals. However, in the case of zinc and copper, their utility is limited to assessing bioavailability because rapid depuration of these metals renders them less useful for long-term environmental monitoring programs. The literature reporting heavy metal impacts on freshwater crayfish, with reference to accumulation levels, is reviewed and summarized with respect to their suitability as bioindicators. Summarized published data from unpolluted or control localities can be used as referential values in crayfish, and consequently help with evaluation of monitored sites.     

A Modeling Study of the Impact of a Power Plant on Ground-Level Ozone in Relation to its Location: Southwestern Spain as a Case Study

The impact of atmospheric industrial emissions on secondary pollutant formation depends on many factors; one of the most important being the environmental setting in which the industry is located. The environmental setting affects an industry’s impact on ozone levels through both the air mass dispersion (a function of topography and wind patterns) and the emissions of organic volatile compounds (VOC) and nitrogen oxides (NO _x ) in the area. This model-based study shows how the sensitivity of surface ozone changes with the choice of source location. For the analysis, seven points distributed along the Tinto–Guadalquivir Basin (in the Southwestern Iberian Peninsula) were selected. This area is characterized by the close proximity of natural environments and crop fields to cities, roads, and industrial areas with high NO _x emissions. Natural VOC emissions represent more than 60% of the total non-methane volatile organic compounds emitted in the study area. The results reveal that the largest increases in ozone levels are produced when the industry is located both far away from NO _x emission sources and near to biogenic VOC emissions. Furthermore, the highest increases over the hourly and 8-hourly maximums, as well as the highest accumulated daily values, are found in areas characterized by high VOC/NO _x emission ratios and NO _x sensitivity. The study of the recurrent meteorological patterns along with the distribution of chemical indicators of the O₃–NO _x –VOC sensitivity allows the determination of the industry’s geographical impact on ozone levels. This information enables air quality managers to decide the future location of an industry minimizing its impact on smog levels.     

In Situ Heavy Metal Accumulation in Lettuce Growing Near a Former Mining Waste Disposal Area: Implications for Agricultural Management

Mining wastes may pose risk nearby urban and agricultural areas. We investigated a lettuce crop land close to a former capped mine tailing in order to determinate the metal uptake by crops. Soil plot sampling design within the crop area and two transects along the tailing were performed. In addition, lettuces (root and leaves) were analyzed after transplant and harvest. The results showed a pH of around 7–8 for all the soil samples. Total metal concentrations were as follows: 190–510 mg kg^?1 Pb, 13–21 mg kg^?1 Cu, and 210–910 mg kg^?1 Zn. Diethylene triamine pentaacetic acid-extractable Pb was around 18% of the total Pb in some samples. Transects along the base and on the plateau of the tailing showed high metal concentrations of Pb (up to 5,800 mg kg^?1) and Zn (up to 4,500 mg kg^?1), indicating that capping layer had been eroded. Lettuce leaves showed Pb concentrations within standard for human health (<0.3 mg kg^?1 in fresh weight). For essential micronutrients such as Cu and Zn, leaves had optimal content (10–28 mg kg^?1 Cu, 60–85 mg kg^?1 Zn). A continued monitoring in metal uptake is needed in crop lands close to mining wastes in order to prevent risks in food safety. Capped tailings must be monitored and rehabilitation works performed from time to time.