Bioindicator reliability: the example of Bel W3 tobacco (Nicotiana tabacum L.)

The use of the Bel W3 tobacco variety revealed the existence of an intrapopulation variation of sensitivity to the ozone effect leading to the isolation of two categories of plants: sensitive and resistant. The use of sensitive plants revealed the existence of a biological ozone efficiency threshold at a level close to 34.5 ppb. Under the threshold, the biological effects observed were weak or very weak. They can be attributed to the occurrence of secondary phenomena (chemical, physiological, pathological, etc.), inducing or inhibiting necrosis, which become perceptible here.     

Growth of soybean at future tropospheric ozone concentrations decreases canopy evapotranspiration and soil water depletion

Tropospheric ozone is increasing in many agricultural regions resulting in decreased stomatal conductance and overall biomass of sensitive crop species. These physiological effects of ozone forecast changes in evapotranspiration and thus in the terrestrial hydrological cycle, particularly in intercontinental interiors. Soybean plots were fumigated with ozone to achieve concentrations above ambient levels over five growing seasons in open-air field conditions. Mean season increases in ozone concentrations ([O₃]) varied between growing seasons from 22 to 37% above background concentrations. The objective of this experiment was to examine the effects of future [O₃] on crop ecosystem energy fluxes and water use. Elevated [O₃] caused decreases in canopy evapotranspiration resulting in decreased water use by as much as 15% in high ozone years and decreased soil water removal. In addition, ozone treatment resulted in increased sensible heat flux in all years indicative of day-time increase in canopy temperature of up to 0.7 °C.     

Lichen microalgae are sensitive to environmental concentrations of atrazine

The identification of new organisms for environmental toxicology bioassays is currently a priority, since these tools are strongly limited by the ecological relevance of taxa used to study global change. Lichens are sensitive bioindicators of air quality and their microalgae are an untapped source for new low-cost miniaturized bioassays with ecological importance. In order to increase the availability of a wider range of taxa for bioassays, the sensitivity of two symbiotic lichen microalgae, Asterochloris erici and Trebouxia sp. TR9, to atrazine was evaluated. To achieve this goal, axenic cultures of these phycobionts in suspension were exposed to a range of environmental concentrations of the herbicide atrazine, a common water pollutant. Optical density and chlorophyll autofluorescence were used as endpoints of ecotoxicity and ecophysiology on cell suspensions. Results show that lichen microalgae show high sensitivity to very low doses of atrazine, being higher in Asterochloris erici than in Trebouxia sp. TR9. We conclude that environmental concentrations of atrazine could modify population dynamics probably through a shift in reproduction strategies of these organisms. This seminal work is a breakthrough in the use of lichen microalgae in the assessment of micropollution effects on biodiversity.     

Critical levels for ozone effects on vegetation in Europe

The evidence of detrimental effects of ozone on vegetation in Europe, and the need to develop international control policies to reduce ozone exposures which are based on the effects of the pollutant, has led to attempts to define so-called critical levels of ozone above which adverse effects on trees, crops and natural vegetation may occur. This review is a critical assessment of the scientific basis of the concepts used to define critical levels for ozone and identifies the key limitations and uncertainties involved. The review focuses on the Level I critical level approach, which provides an environmental standard or threshold to minimise the effects of ozone on sensitive receptors, but does not seek to quantify the impacts of exceeding the critical level under field conditions. The concept of using the AOT (accumulated exposure over a threshold) to define long-term ozone exposure is demonstrated to be appropriate for several economically important species. The use of 40 ppb (giving the AOT40 index) as a threshold concentration gives a good linear fit to experimental data from open-top chambers for arable crops, but it is less certain that it provides the best fit to data for trees or semi-natural communities. Major uncertainties in defining critical level values relate to the choice of response parameter and species; the absence of data for many receptors, especially those of Mediterranean areas; and extrapolation to field conditions from relatively short-term open-top chamber experiments. The derivation of critical levels for long-lived organisms, such as forest trees, may require the use of modelling techniques based on physiological data from experimental studies. The exposure-response data which have been applied to derive critical levels should not be used to estimate the impacts of ozone over large areas, because of the uncertainties associated with extrapolation from the open-top chamber method, especially for forest trees, and because of spatial variation in atmospheric and environmental conditions, which may alter ozone uptake.     

Response to ambient ozone of two white clover (Trifolium repens L.cv. "Regal") clones, one resistant and one sensitive, grown in a Mediterranean environment

Two clones of white clover (Trifolium repens L.) differing in ozone tolerance were grown in southern Italy during 1997 and 1998 to study the effects of ambient ozone exposure on yield, leaf morphology and water use. Ambient ozone levels were high in both years with values exceeding the threshold for leaf injury reported in the literature. In both years ozone injury was observed on the sensitive clone (NC-S) but not on the resistant one (NC-R), and leaf and stolon dry matter production was significantly lower in NC-S than in NC-R. However, it cannot be excluded that other factors, such as high temperature, interacted with the effect of ozone on biomass production. The clones differed in morphological characteristics. Lower total leaf area in NC-S plants was due to a smaller number of leaves per plant, but the average area per leaf was higher in NC-S. Specific leaf weight and net assimilation rate were higher in the more productive clone (NC-R). Cumulative plant water use was higher in NC-R in each growth period because of the larger leaf area; by contrast, water use per unit leaf area was higher in NC-S, indicating higher leaf conductance to water vapour. The results suggest that ozone significantly reduces the yield of sensitive white clover plants under well-watered conditions, and that the difference in ozone tolerance between clover clones is related to differences in leaf morphology and water use.     

Study on the formation and transport of ozone in relation to the air quality management and vegetation protection in Tenerife (Canary Islands)

An experimental study on the formation and transport of ozone in ambient air was performed in Tenerife (Canary Islands) in order to investigate the processes affecting ozone levels and air quality. The special features of Tenerife (prevalence of the trade wind pattern (NE), orography and the specific location of the local ozone sources) permit to quantify the role of the ‘long-range transport from northern latitudes' versus the ‘formation and transport of ozone downwind of the main urban areas' of Tenerife. Levels of O₃, NO₂ and O_X were monitored in different types of environments to achieve this purpose. The results showed that: (1) upwind of the urban areas ozone is mainly transported from the ocean by trade winds, (2) local ozone titration (by NO) and ozone replenishment from the ocean are the main causes of ozone variations in urban and suburban areas, and (3) photochemical ozone production occurs downwind of the urban areas. Photochemical production causes daylight O₃ and O_X levels downwind of urban areas to be frequently (60% and 35% days/year, respectively) higher than upwind of the urban sites (O₃ and O_X excess frequently in the range 5–20 ppbv). Due to the above processes, different daily ozone cycles occur in short distances (<30 km), with maximum O₃ levels during daylight or night depending on the site. Ozone phytotoxicity was assessed by calculating the AOT40 index upwind and downwind of the main urban areas. The critical value for the 5-day-AOT40 index was simultaneously exceeded at the two sites (few times/year) during long-range transport events. During the additional exceedances of the critical value downwind of the urban area, relatively high 5-day-AOT40 values were recorded upwind of the urban site. Thus, long-range transport from northern latitudes may produce relatively high 5-days-AOT40 levels in the oceanic boundary layer. These results are important for the protection of the large number of endemic plants in the Canaries. The conceptual model discussed in this study may be qualitatively applied to other islands which possess features similar to those of Tenerife.     

Instrumental recording and biomonitoring of ambient ozone in the Greek countryside

Among eight commercial Greek varieties of tobacco (Nicotiana tabacum L.) tested for their ozone-sensitivity levels, the Zichnomirodata (KK6/5) variety was found to be the most sensitive, although less sensitive than the well-known super-sensitive Bel-W3. Besides qualitative differences in the appearance of macroscopic symptoms these two varieties can be used simultaneously as a reliable pair of ozone bioindicators. The occurrence of ozone in the Greek countryside was surveyed by biomonitoring in 14 rural regions over the country and by a simultaneous biomonitoring and instrumental recording of ozone concentrations at a single remote side (Pournaria, Arcadia). Phytotoxic symptoms were observed mainly on the leaves of Bel-W3 and occasionally on those of Zichnomirodata varieties, suggesting that ozone levels were high enough to affect at least sensitive species. The instrumental monitoring (during a total period of 912 h) revealed maximum hourly O3 concentration 62 ppb, while the thresholds of 30, 40 and 50 ppb were exceeded for 40%, 20% and 6% of the recording period, respectively. The accumulated exposure over 40 ppb (AOT40) for the daylight hours over the 38 monitored days was 680 ppb h.     

A synthesis of AOT40-based response functions and critical levels of ozone for agricultural and horticultural crops

Crop-response data from over 700 published papers and conference proceedings have been analysed with the aim of establishing ozone dose-response functions for a wide range of European agricultural and horticultural crops. Data that met rigorous selection criteria (e.g. field-based, ozone concentrations within European range, full season exposure period) were used to derive AOT40-yield response functions for 19 crops by first converting the published ozone concentration data into AOT40 (AOT40 is the hourly mean ozone concentration accumulated over a threshold ozone concentration of 40 ppb during daylight hours, units ppm h). For any individual crop, there were no significant differences in the linear response functions derived for experiments conducted in the USA or Europe, or for individual cultivars. Three statistically independent groups were identified: ozone sensitive crops (wheat, water melon, pulses, cotton, turnip, tomato, onion, soybean and lettuce); moderately sensitive crops (sugar beet, potato, oilseed rape, tobacco, rice, maize, grape and broccoli) and ozone resistant (barley and fruit represented by plum and strawberry). Critical levels of a 3 month AOT40 of 3 ppm h and a 3.5 month AOT40 of 6 ppm h were derived from the functions for wheat and tomato, respectively.     

Determination of coplanar and ortho substituted PCBs in some sewage sludges of Switzerland using HRGC/ECD and HRGC/MSD

The three most toxic coplanar PCB congeners PCB 77, 126 and 169 as well as 7 mono-and di-ortho-PCBs (PCB 28, 52, 101, 118, 153, 138, 180) were measured in 19 sewage sludges. Concentrations of the coplanar PCBs ranged between 231–5050 ng/kg dw for PCB 77, 43–1511 ng/kg dw for PCB 126 and 29–273 ng/kg dw for PCB 169. Comparable results were obtained with mass selective detection in the single ion monitoring mode (MSD-SIM) except for PCB 169, where MSD-SIM was less sensitive than ECD. The sum of the 7 mono-and di-ortho-PCBs, which are routinely measured as representatives of the PCB fraction, reached levels between 43–550 μg/kg dw and agreed well with results previously obtained in our laboratory. Based on our data, a selective accumulation of the coplanar PCBs could not be observed. Sludges that received industrial effluents clearly showed higher PCB levels than rural ones. Finally, calculation of TEQ equivalents of the coplanar PCBs in the sewage sludges revealed that, based on application rates normally used in Switzerland, the contribution of the sludges to soil pollution seems to be of minor importance.     

Ammonium and nitrate tolerance in lichens

Since lichens lack roots and take up water, solutes and gases over the entire thallus surface, these organisms respond more sensitively to changes in atmospheric purity than vascular plants. After centuries where effects of sulphur dioxide and acidity were in the focus of research on atmospheric chemistry and lichens, recently the globally increased levels of ammonia and nitrate increasingly affect lichen vegetation and gave rise to intense research on the tolerance of lichens to nitrogen pollution. The present paper discusses the main findings on the uptake of ammonia and nitrate in the lichen symbiosis and to the tolerance of lichens to eutrophication. Ammonia and nitrate are both efficiently taken up under ambient conditions. The tolerance to high nitrogen levels depends, among others, on the capability of the photobiont to provide sufficient amounts of carbon skeletons for ammonia assimilation. Lowly productive lichens are apparently predisposed to be sensitive to excess nitrogen.     

Effects of ozone fumigation on epiphytic macrolichens: ultrastructure, CO2 gas exchange and chlorophyll fluorescence

The lichen species Anaptychia ciliaris, Collema nigrescens, Evernia prunastri, Hypogymnia bitteri, Lobaria pulmonaria, Pseudevernia furfuracea and Usnea rigida s.l. were fumigated with site-relevant concentrations (for Central Europe) of ozone over 80 days (180 microg m(-3) during daytime, 80 microg m(-3) during the night). Chlorophyll fluorescence measurements revealed a significant reduction of Fv/Fm after ozone fumigation in five of the species investigated, indicating severe stress on photosystem II due to ozone. The physiological impairment paralleled our fine structural investigations, revealing a significantly higher percentage of collapsed photobiont cells. This indicates that the effects of ambient ozone concentrations under experimental conditions included biophysical and physiological, as well as structural impairment in the lichens studied.     

Using plant biomonitors and flux modelling to develop O3 dose-response relationships in Catalonia

We used tobacco Bel-W3 biomonitoring data and ozone flux modelling (WINDEP model) with the aim of developing the absorbed dose-response relationship, and comparing this approach with the most commonly used AOT40 (the sum of hourly ozone concentrations above a cut-off of 40 ppb during daylight hours, when global radiation exceeds 50 W m(-2)) in the estimation of exposure-damage curves. Leaf damage values were more related to OAD(15 days, potential) (potential ozone absorbed dose calculated over 15 consecutive days) than to AOT40 in all the studied stations. An OAD(15 days, potential) of 180 mg m(-2) was found to be the threshold for damage to the most sensitive species in this region under well watered conditions. The results show the applicability of the flux approach for risk assessment at the local scale, the improvement of the ozone damage estimation when the potential absorbed dose is modelled and used instead of just the ozone exposure, and finally, the possibilities opened by the use of biomonitoring networks.     

Relationship between epiphytic lichens, trace elements and gaseous atmospheric pollutants

A study was conducted to determine the joint effect of gaseous atmospheric pollutants and trace elements on epiphytic lichens. We used our data to test the hypothesis that lichens are generally insensitive to toxic effects of trace elements, and can therefore be used as accumulator organisms to estimate concentrations of these elements in the environment. In a field study in The Netherlands the abundance of epiphytic lichen species was estimated, and their supporting bark was collected. Concentrations of a range of trace elements were determined in the bark, and concentrations of atmospheric trace gases were estimated at the sites of collection. Multivariate statistics were used to determine the relation between the abundance of the species and pollutant concentrations. Atmospheric SO2 and NO2 appeared to be the most important factors determining lichen biodiversity. Nearly all species were sensitive to these compounds. The effect of the other trace elements was very slight; only Sb had a significantly negative effect on the abundance of a few species. It is concluded that lichens can safely be used as accumulator organisms in pollution studies, provided that concentration in lichen thalli reflect atmospheric concentrations.     

Growth and premature leaf fall in American aspen as bioindications for ozone

Several clones of the American aspen (Populus tremuloides Michx.) differing in ozone sensitivity were fumigated continuously for 105 days with different ozone concentrations, producing different ozone doses. The suitability of the clones as possible replacements for tobacco as bioindicators in forested areas was tested. The life span of leaves of the ozone-sensitive clones was distinctly reduced by ozone. The difference to controls was significant even in the relatively clean air at Birmensdorf. Growth was diminished, too, but the decrease in height was much less pronounced than that in weight. In sensitive clones an after-effect was detectable, since the weight of new shoots (grown from reserves) decreased with increasing ozone stress in the previous year. Since the tolerant clone grew less, it is-at first sight-of decreased economic interest from the point of view of rapid productivity. In the long run, however, it might be economically interesting because it may be able to maintain productivity (although at a lower level) due to better competitive strength.     

On the decline of ground lichen forests in the Swedish boreal landscape: Implications for reindeer husbandry and sustainable forest management

Lichens are a bottleneck resource for circumpolar populations of reindeer, and as such, for reindeer husbandry as an indigenous Sami land-use tradition in northern Sweden. This study uses ground lichen data and forest information collected within the Swedish National Forest Inventory since 1953, on the scale of northern Sweden. We found a 71 % decline in the area of lichen-abundant forests over the last 60 years. A decline was observed in all regions and age classes and especially coincided with a decrease of >60 year old, open pine forests, which was the primary explanatory factor in our model. The effects of reindeer numbers were inconclusive in explaining the decrease in lichen-abundant forest. The role that forestry has played in causing this decline can be debated, but forestry can have a significant role in reversing the trend and improving ground lichen conditions.

Electronic supplementary material

The online version of this article (doi:10.1007/s13280-015-0759-0) contains supplementary material, which is available to authorized users.     

Effect of Nitrogen Nutrition on the Response of Tobacco to Ozone in the Atmosphere

Experiments were conducted to determine the effect of nitrogen nutrition on the response of tobacco to ozone in the atmosphere. With Nicotiana rustica L., a noncommercial tobacco, plants receiving an “optimum” nitrogen supply were found to be more susceptible to ozone injury than those receiving deficient or luxury supplies. Furthermore, it was found on analyses of the third oldest tobacco leaf that, while these “optimum” plants showed little or no increase in respiration as a result of ozone in concentrations too low to produce visible injury, plants grown with deficient or excess nitrogen showed a significant increase in respiration under these same conditions. The increase in respiration was accompanied by a comparable and equally significant decrease in carbohydrate con-lent in the latter plants which was not apparent in plants at the “optimum” nitrogen supply, as well as an increase in total nitrogen, protein-nitrogen, and amino-acid nitrogen. In conclusion, it is suggested that the carbohydrate reserve built up in plants grown with deficient nitrogen nutrition serves as protection against the ozidative potential of ozone. Likewise, a decrease in carbohydrate content below that of “optimum” plants provides protection in plants grown with an overabundance of nitrogen fertilization, though possibly by a more complex mechanism.     

Effects of atmospheric pollution on lichen biodiversity (LB) in a Mediterranean region (Liguria, northwest Italy)

The effects of air pollution on lichen biodiversity (LB) were monitored in Liguria (northwest Italy). A systematic sampling strategy was adopted in order to avoid the influence of spatial autocorrelation on the results. An eight LB class scale permitted to point out the levels of naturality/alteration in the region. The comparison of these results with the ones obtained by mean of physico-chemical methodologies shows a good accordance. The results of this study suggest the possibility of designing an integrated monitoring network, in which biological monitoring will allow to estimate the level of alteration in remote areas, which account for most of this region and which are currently not covered by measurements with automatic systems.     

Lichens as sentinels for air pollution at remote alpine areas (Italy)

The present study was undertaken with the aim of using epiphytic lichens as sentinels for air pollution at two remote alpine sites (1,400 and 1,800 m above sea level (asl)) of NW Italy. The results indicated that the site at 1,800 m prompted for early warning indications of biological changes. Although levels of the many elements assayed in samples of the lichen Pseudevernia furfuracea (L.) Zopf, ranging from minor elements (e.g., Al) to ultra-trace (e.g., Pt), were at normal levels, indications of a slowly worsening environment were given by the lichen biodiversity and by damage to cell membranes. The analysis of Pb isotopic ratios suggested that the origin of Pb accumulated in lichens is not local, but linked to the long-range transport by air masses. It was concluded that the origin of pollutants is from air mass coming from the Po plain of Italy and from densely populated areas of Switzerland and France.     

Tidal salt marsh sediment in California, USA: part 3. Current and historic toxicity potential of contaminants and their bioaccumulation

To assess potential health risks to benthic organisms from exposure to toxic contaminants, sediment chemistry data from five salt marshes along the coast of California were compared with threshold effects levels (TELs) and probable effects levels (PELs). As an integrated estimate of toxicity potential of multiple contaminants, mean PEL quotients (mPELQs) were used to categorize sampling stations into three groups: high (>0.5), medium (0.1–0.5) and low (<0.1). In all sediments from Stege Marsh located in San Francisco Bay, at least one contaminant exceeded PELs by up to 18-fold and mPELQs were higher than 0.7. Mean PELQs in two core sediments from eastern Stege Marsh ranged from 0.7 to 2.1, indicating that benthic organisms in Stege Marsh may have been adversely affected for several decades. To investigate bioavailability and bioaccumulation of contaminants in sediments, longjaw mudsuckers (Gillichthys mirabilis) were transplanted to six Stege Marsh stations for 60 days. Body burdens of organic contaminants clearly showed that they were readily available for benthic organisms. Measured concentrations of organic contaminants in mudsuckers were similar to estimated levels computed using a theoretical bioaccumulation potential model. Levels of PCBs and arsenic in mudsuckers were higher than screening values set as guidelines for the protection of humans and levels of PCBs and DDTs were higher than criteria for wildlife. The results of this study indicate that the levels of contaminants in Stege Marsh sediments may not fully support the well-being of benthic organisms and also may provoke adverse effects on fish-eating animals and humans through trophic transfer.