Two holm oak (Quercus ilex L.) sites in Tuscany, Central Italy, were studied in July 2000. Leaf morphological characteristics (area, dry weight, specific dry weight, water content, epicuticular wax amount, stomatal density) and leaf fluorescence were measured. Ozone flux into the leaves was calculated on a stomatal conductance basis. Among the measured parameters, only stomatal density and wax amount significantly differed between the sites. This response might indicate an adaptation to air pollutants, of which tropospheric ozone might be of the greatest importance

Tropospheric ozone is a dangerous atmospheric pollutant for forest ecosystems when it penetrates stomata. Thresholds for ozone-risk assessment are based on accumulated stomatal ozone fluxes such as the Phytotoxic Ozone Dose (POD). In order to identify the effect of ozone on a Holm oak forest in central Italy, four flux-based ozone impact response functions were implemented and tested in a multi-layer canopy model AIRTREE and evaluated against Gross Primary Productivity (GPP) obtained from observations of Eddy Covariance fluxes of CO₂. To evaluate if a clear phytotoxic threshold exists and if it changes during the year, six different detoxifying thresholds ranging between 0 and 5 nmol O₃ m⁻² s⁻¹ were tested.The use of species-specific rather than more general response functions based on plant functional types (PFT) increased model accuracy (RMSE reduced by up to 8.5%). In the case of linear response functions, a threshold of 1 nmol m⁻² s⁻² produced the best results for simulations of the whole year, although the tolerance to ozone changed seasonally, with higher tolerance (5 nmol m⁻² s⁻¹ or no ozone impact) for Winter and Spring and lower thresholds in Summer and Fall (0–1 nmol m⁻² s⁻¹). A “dynamic threshold” obtained by extracting the best daily threshold values from a range of different simulations helped reduce model overestimation of GPP by 213 g C m⁻² y⁻¹ and reduce RMSE up to 7.7%. Finally, a nonlinear ozone correction based on manipulative experiments produced the best results when no detoxifying threshold was applied (0 nmol O₃ m⁻² s⁻¹), suggesting that nonlinear functions fully account for ozone detoxification. The evidence of seasonal changes in ozone tolerance points to the need for seasonal thresholds to predict ozone damage and highlights the importance of performing more species-specific manipulative experiments to derive response functions for a broad range of plant species.

Polycyclic aromatic hydrocarbons (PAHs) are hazardous organic compounds with mutagenic, genotoxic and carcinogenic properties. Although PAHs in soil can cause toxicity to microorganisms, the microbial community is able to degrade these compounds. For this reason, it is important to study acute and short-term effects of PAH contamination on soil microbial community, also to shed light on its possible exploitation in soil restoration.The effects of acute PAH contamination on the structure and metabolic activity of microbial communities in three forest (beech, holm oak, black pine) soils were studied. The soils were spiked with phenanthrene, pyrene or benzo[a]pyrene and incubated in experimental mesocosms, under controlled conditions. Enzymatic activities (laccase, total peroxidase and hydrolase), as well as microbial biomass and community structure (through phospholipid fatty acid and ergosterol analyses), were evaluated in the three soil systems 4 days after contamination and compared to no-spiked soils. In soil under holm oak, there was a stimulation of Gram+ bacteria after contamination with all the 3 PAHs, whereas in soil under pine, pyrene and phenanthrene additions mainly stimulated fungi and actinomycetes.

Deposition of dissolved organic nitrogen (DON) in both bulk precipitation (BD) and canopy throughfall (TF) has been measured for the first time in the western Mediterranean. The study was carried out over a year from 2012 to 2013 at four evergreen holm oak forests located in the Iberian Peninsula: two sites in the Province of Barcelona (Northeastern Spain), one in the Province of Madrid (central Spain) and the fourth in the Province of Navarra (Northern Spain). In BD the annual volume weighted mean (VWM) concentration of DON ranged from 0.25 mg l−1 in Madrid to 1.14 mg l−1 in Navarra, whereas in TF it ranged from 0.93 mg l−1 in Barcelona to 1.98 mg l−1 in Madrid. The contribution of DON to total nitrogen deposition varied from 34% to 56% in BD in Barcelona and Navarra respectively, and from 38% in Barcelona to 72% in Madrid in TF. Agricultural activities and pollutants generated in metropolitan areas were identified as potential anthropogenic sources of DON at the study sites. Moreover, canopy uptake of DON in Navarra was found in spring and autumn, showing that organic nitrogen may be a supplementary nutrient for Mediterranean forests, assuming that a portion of the nitrogen taken up is assimilated during biologically active periods.

Photosynthetic stimulation and stomatal conductance (Gs) depression in Quercus ilex leaves at a CO2 spring suggested no down-regulation. The insensitivity of Gs to a CO2 increase (from ambient 1500 to 2000 μmol mol-1) suggested stomatal acclimation. Both responses are likely adaptations to the special environment of CO2 springs. At the CO2-enriched site, not at the control site, photosynthesis decreased 9% in leaves exposed to 2x ambient O3 concentrations in branch enclosures, compared to controls in charcoal-filtered air. The stomatal density reduction at high CO2 was one-third lower than the concomitant Gs reduction, so that the O3 uptake per single stoma was lower than at ambient CO2. No significant variation in monoterpene emission was measured. Higher trichome and mesophyll density were recorded at the CO2-enriched site, accounting for lower O3 sensitivity. A long-term exposure to H2S, reflected by higher foliar S-content, and CO2 might depress the antioxidant capacity of leaves close to the vent and increase their O3 sensitivity. Very high CO2 concentrations did not compensate for the effects of O3 on holm oak photosynthesis.

We conducted a field drought manipulation experiment in an evergreen oak Mediterranean forest from 1999 to 2005 to investigate the effects of the increased drought predicted for the next decades on the accumulation of trace elements that can be toxic for animals, in stand biomass, litter and soil. Drought increased concentrations of As, Cd, Ni, Pb and Cr in roots of the dominant tree species, Quercus ilex, and leaf Cd concentrations in Arbutus unedo and of Phillyrea latifolia codominant shrubs. The increased concentration of As and Cd can aggravate the toxic capacity of those two elements, which are already next or within the levels that have been shown to be toxic for herbivores. The study also showed a great reduction in Pb biomass content (100-135 g ha-1) during the studied period (1999-2005) showing the effectiveness of the law that prohibited leaded fuel after 2001. The results also indicate that drought increases the exportation of some trace elements to continental waters. Drought increased biomass concentrations of As and Cd and favors exportation of some trace elements to continental waters in a Mediterranean forest.

In Mediterranean areas, dry deposition is a major component of the total atmospheric N input to natural habitats, particularly to forest ecosystems. An innovative approach, combining the empirical inferential method (EIM) for surface deposition of NO₃⁻ and NH₄⁺ with stomatal uptake of NH₃, HNO₃ and NO₂ derived from the DO₃SE (Deposition of Ozone and Stomatal Exchange) model, was used to estimate total dry deposition of inorganic N air pollutants in four holm oak forests under Mediterranean conditions in Spain. The estimated total deposition varied among the sites and matched the geographical patterns previously found in model estimates: higher deposition was determined at the northern site (28.9 kg N ha⁻¹ year⁻¹) and at the northeastern sites (17.8 and 12.5 kg N ha⁻¹ year⁻¹) than at the central-Spain site (9.4 kg N ha⁻¹ year⁻¹). On average, the estimated dry deposition of atmospheric N represented 77% ± 2% of the total deposition of N, of which surface deposition of gaseous and particulate atmospheric N averaged 10.0 ± 2.9 kg N ha⁻¹ year⁻¹ for the four sites (58% of the total deposition), and stomatal deposition of N gases averaged 3.3 ± 0.8 kg N ha⁻¹ year⁻¹ (19% of the total deposition). Deposition of atmospheric inorganic N was dominated by the surface deposition of oxidized N in all the forests (means of 54% and 42% of the dry and total deposition, respectively). The relative contribution of NO₂ to dry deposition averaged from 19% in the peri-urban forests to 11% in the most natural site. During the monitoring period, the empirical critical loads provisionally proposed for ecosystem protection (10–20 kg N ha⁻¹ year⁻¹) was exceeded in three of the four studied forests.

Atmospheric nitrogen deposition is one of the main threats for biodiversity and ecosystem functioning. Measurement techniques like ion-exchange resin collectors (IECs), which are less expensive and time-consuming than conventional methods, are gaining relevance in the study of atmospheric deposition and are recommended to expand monitoring networks. In the present work, bulk and throughfall deposition of inorganic nitrogen were monitored in three different holm oak forests in Spain during two years. The results obtained with IECs were contrasted with a conventional technique using bottle collectors and with a literature review of similar studies. The performance of IECs in comparison with the conventional method was good for measuring bulk deposition of nitrate and acceptable for ammonium and total dissolved inorganic nitrogen. Mean annual bulk deposition of inorganic nitrogen ranged 3.09–5.43 kg N ha−1 according to IEC methodology, and 2.42–6.83 kg N ha−1 y−1 using the conventional method. Intra-annual variability of the net throughfall deposition of nitrogen measured with the conventional method revealed the existence of input pulses of nitrogen into the forest soil after dry periods, presumably originated from the washing of dry deposition accumulated in the canopy. Important methodological recommendations on the IEC method and discussed, compiled and summarized.