Degradation of oxytetracycline and doxycycline by ozonation: Degradation pathways and toxicity assessment.

Tetracyclines are one of the antibiotics widely employed worldwide and frequently detected in surface waters because of incomplete removal from wastewater treatment. Various advanced oxidation processes have been investigated for tetracyclines degradation and their transformation products (TPs) have recently gained more attention. Studies on ozonation are however seldom for the degradation of oxytetracycline (OTC) and doxycycline (DTC). In the present study, a lower O3 inlet gas concentration (4.67 ± 0.13 mg/L), supplied at a flow rate of 0.27 L/min, was shown to be more effective at removing OTC than the same dose of ozone applied at higher inlet gas concentration (up to 6.29 mg/L) over a shorter time at the same flow rate. The use of pCBA and t-BuOH indicated that ozone plays a more important role in the degradation of OTC than HO•. The DTC degradation was less efficient than for OTC, with 99 % removal requiring twice the amount of ozone. OTC had almost no inhibition of Vibrio fischeri, however, the inhibition ratio was increased to 37 % (5-min) and 46 % (15-min) within 1 min of ozonation. Contrastly, DTC had toxic effects on V. fischeri (inhibition rate5min of 84 %) and sustained toxicity in samples treated for up to 40-min. The observed toxicities after treatment could be explained by the identified TPs (26 TPs for OTC and 23 for DTC, some identified for the first time) and their quantitative structure-activity relationship analysis data. Several TPs showed toxic or extremely toxic predicted effects on fish, daphnid, and green algae, corresponding with the V. fischeri inhibition results. Among the possible degradation pathways, aromatic ring hydroxylation and ring-opening pathways could lead to the formation of TPs less harmful to the environment.

A Practical Assessment of the Disinfectant Efficacy of UV Light with and without Ozone Using a Novel Transfer Hatch in a Research Animal Facility.

Most in vivo animal research and breeding using mice and rats in China takes place in facilities under barrier conditions. Items being moved across the barrier are typically disinfected using UV radiation in a transfer hatch. However, the time periods necessary for this disinfection technique are inefficient, and disinfection is frequently incomplete, especially if concealed surfaces are present. The current study used a newly developed transfer hatch incorporating both UV and ozone disinfection to examine disinfection efficacy against 4 bacteria species (Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii). Disinfection trials used UV and ozone, applied separately and in combination, for up to 30 min. Separate and combined treatments were also tested with a UV barrier. We found that if UV radiation has direct contact with surfaces, it is an efficient disinfection method. However, where surfaces are concealed by a UV barrier, UV radiation performs relatively poorly. The results of this study indicate that a combination of UV and ozone produces the most effective disinfection and is markedly quicker than current disinfection times for UV applied on its own. This novel transfer hatch design therefore allows more complete and efficient disinfection, improves workflow, and reduces barrier breaches by pathogens that may affect animal health and welfare and compromise research outcomes.

An assessment of growth, floral morphology, and metabolites of a medicinal plant Sida cordifolia L. under the influence of elevated ozone.

Tropospheric ozone (O3) is a major secondary air pollutant and greenhouse gas, and its impact on growth, yield, and its quality is well established in the case of crop plants. However, the effects of tropospheric O3 have not been comprehensively studied on medicinal plants. Therefore, a field study was planned on a medicinally important Sida cordifolia L. plant (commonly known as country mallow or Bala) to assess the expected changes on the morphology, growth, and leaf injury under elevated O3 (ambient + 20 ppb) by using open-top chambers (OTCs) at 30, 60, and 90 days after treatment (DAT), while leaf and root metabolites were observed at 60 DAT. At all the growth stages, significant leaf damage was recorded as foliar injury symptoms. Most of the growth parameters also showed significant reductions at all the growth stages. Plants under elevated O3 showed a significant negative impact on most of the reproductive parts of the plant. Leaf weight ratio (LWR) showed significant increment at early stages while reduced at 90 DAT; however, root shoot ratio (RSR) showed a significant reduction at 60 DAT. The majority of the steroid metabolites showed an increase in root and leaves under elevated O3, while terpenes showed variable response. Due to O3 stress, most of the major metabolites showed an increase possibly due to their role in defense and other metabolic activities. Based on the outcomes, it is concluded that the future increase in the levels of tropospheric O3 will impact a significant effect on important metabolites of medicinal plants growing in tropical countries like India.

Diversity Assessment of Toxic Cyanobacterial Blooms during Oxidation.

Fresh-water sources of drinking water are experiencing toxic cyanobacterial blooms more frequently. Chemical oxidation is a common approach to treat cyanobacteria and their toxins. This study systematically investigates the bacterial/cyanobacterial community following chemical oxidation (Cl2, KMnO4, O3, H2O2) using high throughput sequencing. Raw water results from high throughput sequencing show that Proteobacteria, Actinobacteria, Cyanobacteria and Bacteroidetes were the most abundant phyla. Dolichospermum, Synechococcus, Microcystis and Nostoc were the most dominant genera. In terms of species, Dolichospermum sp.90 and Microcystis aeruginosa were the most abundant species at the beginning and end of the sampling, respectively. A comparison between the results of high throughput sequencing and taxonomic cell counts highlighted the robustness of high throughput sequencing to thoroughly reveal a wide diversity of bacterial and cyanobacterial communities. Principal component analysis of the oxidation samples results showed a progressive shift in the composition of bacterial/cyanobacterial communities following soft-chlorination with increasing common exposure units (CTs) (0-3.8 mg·min/L). Close cyanobacterial community composition (Dolichospermum dominant genus) was observed following low chlorine and mid-KMnO4 (287.7 mg·min/L) exposure. Our results showed that some toxin producing species may persist after oxidation whether they were dominant species or not. Relative persistence of Dolichospermum sp.90 was observed following soft-chlorination (0.2-0.6 mg/L) and permanganate (5 mg/L) oxidation with increasing oxidant exposure. Pre-oxidation using H2O2 (10 mg/L and one day contact time) caused a clear decrease in the relative abundance of all the taxa and some species including the toxin producing taxa. These observations suggest selectivity of H2O2 to provide an efficient barrier against toxin producing cyanobacteria entering a water treatment plant.

Cost-effective culture of human induced pluripotent stem cells using UV/ozone-modified culture plastics with reduction of cell-adhesive matrix coating.

Human induced pluripotent stem cells (hiPSCs) are considered to be one of the most promising cell resources for regenerative medicine. HiPSCs usually maintain their pluripotency when they are cultured on feeder cell layers or are attached to a cell-adhesive extracellular matrix. In this study, we developed a culture system based on UV/ozone modification for conventional cell culture plastics to generate a suitable surface condition for hiPSCs. Time of flight secondary ion mass spectrometry (ToF-SIMS) was carried out to elucidate the relationship between hiPSC adhesion and UV/ozone irradiation-induced changes to surface chemistry of cell culture plastics. Cell culture plastics with modified surfaces enabled growth of a feeder-free hiPSC culture with markedly reduced cell-adhesive matrix coating. Our cell culture system using UV/ozone-modified cell culture plastics may produce clinically relevant hiPSCs at low costs, and can be easily scaled up in culture systems to produce a large number of hiPSCs.

In vivo toxicity assessment of aflatoxin B1-contaminated corn after ozone degradation.

Corn is an important food and feedstuff in China and worldwide. The problems caused by aflatoxin B1-contaminated corn (ACC) are of great concern. Our previous studies have demonstrated that ozone can effectively degrade AFB1 in corn, prompting us to investigate the in vivo toxicity of treated ACC. In this study, 35 Kunming mice were used to assess the in vivo toxicity of ozone treated ACC. Results indicated that compared to mice fed with basal feedstuff (provided by the Shanghai SLAC Laboratory), those fed with ACC have significantly decreased mean weight as well as total protein (TP), albumin (ALB), and globulin (GLB) contents (p < 0.05). On the other hand, the liver and kidney/body weight ratio as well as the serum alanine transaminase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) levels significantly increased (p < 0.05). Obvious histopathological changes were found in the liver and kidney. When mice were fed with the ozone-treated ACC, no significant differences were observed in the mean weight, the liver and kidney/body weight ratio and in the major serum indexes ALT, TP, ALB, and GLB (p > 0.05). However, AST and ALP significantly increased (p < 0.05), and slight histopathological changes were found in liver tissues. This study indicated that ACC may lead to significant changes in various physiological characteristics and biochemical indexes in liver and kidney tissues, but ozone treatment of ACC could significantly reduce these changes.

Assessment of biofilm formation among clinical isolates of Acinetobacter baumannii in burn wounds in the west of Iran.

Burn wound infection by A. baumannii is one of the predominant cause of mortality worldwide. The present investigation aimed at determination of antimicrobial resistance profile and expression of the biofilm-related genes in A. baumannii isolated from hospitalized patients with burn wound infection in Kermanshah hospitals. Sixty four isolates of A. baumannii were recovered from burn wound of hospitalized patients at hospitals in Kermanshah. The antimicrobial susceptibility testing (AST) was performed. Biofilm formation was measured and antibiotic resistance was compared between before and after of biofilm formation. The polymerase chain reaction (PCR) and Real-Time PCR were performed to detect of abaI and pgaD genes. The biofilm producer isolates and the most resistant isolates were exposed to ozone gas .More than 70% strains were resistance to Erythromycin, Ofloxacin, Ceftazidime, Ceftriaxone, and Ticarcillin-clavulanic acid and 50% isolates were resistant to Imipenem. Thirty one (48.4%) isolates were biofilm producer. The pgaD and abaI genes were positive in 29 (45.3%) and 9 (14%) isolates, respectively. Real time PCR demonstrated that the copy numbers of the pgaD and abaI genes after biofilm formation were increased. After exposure to ozone, biofilm formation reduced in all very strong biofilm producing isolates. Our results showed that after biofilm formation, an increased resistance was observed in most isolates. Also rising expression of abaI gene was associated with biofilm formation and an increase of antibiotic resistance. In the current study, both biofilm formation and antibiotic resistance were reduced after O3 exposure.

In vitro assessment of the recurrent doses of topical gaseous ozone in the removal of Enterococcus faecalis biofilms in root canals.

AIM: To evaluate the potential antibacterial effect of recurrent doses of topical gaseous ozone on the Enterococcus faecalis biofilms0 growth in human root canals in vitro. MATERIALS AND METHODS: One hundred and thirty four human single-rooted mandibular premolars were enlarged to a size 35 K-File. Each root canal were inoculated with an overnight culture of Enterococcus faecalis 0ATCC 29212 in tryptic soy broth for 24 hours and incubated for 7 days at 37°C. At 7-day interval, 4 specimens were prepared for Scanning Electron Microscope (SEM) analysis to confirm the presence and purity of biofilms whilst the other contaminated root canals were irrigated and disinfected. One hundred root canals of total 134 specimens were selected to create the experimental groups and divided into 5 subgroups. In each experimental group ( n = 20) root canals), recurrent ozone doses were applied with different irrigation and disinfection protocols in 5 different time intervals. Bacterial growth was analyzed by counting viable E. faecalis on tryptic soy agar plates. RESULTS: According to intergroup comparison results observed in the final sample collection analysis, the amount of remaining bacteria in the positive control group were found to be significantly higher compared to Groups 1, 2, 3, 4, 5 and the material control group ( P < 0.01). The remaining amount of bacteria in the last count of Group 1 were found to be significantly higher compared to Group 2 ( P < 0.05), Group 4 ( P < 0.01), Group 5 (P < 0.05) and the material control group (P < 0.01). CONCLUSION: The application of topical gaseous ozone in recurrent doses provides a positive effect in the removal of E. faecalis biofilm from root canals. However, during disinfection procedure, the combined use of recurrent doses of topical gaseous ozone with 2% NaOCl enhanced its antibacterial effect against E. faecalis biofilm.

Foliar symptoms triggered by ozone stress in irrigated holm oaks from the city of Madrid, Spain.

BACKGROUND: Despite abatement programs of precursors implemented in many industrialized countries, ozone remains the principal air pollutant throughout the northern hemisphere with background concentrations increasing as a consequence of economic development in former or still emerging countries and present climate change. Some of the highest ozone concentrations are measured in regions with a Mediterranean climate but the effect on the natural vegetation is alleviated by low stomatal uptake and frequent leaf xeromorphy in response to summer drought episodes characteristic of this climate. However, there is a lack of understanding of the respective role of the foliage physiology and leaf xeromorphy on the mechanistic effects of ozone in Mediterranean species. Particularly, evidence about morphological and structural changes in evergreens in response to ozone stress is missing. RESULTS: Our study was started after observing ozone -like injury in foliage of holm oak during the assessment of air pollution mitigation by urban trees throughout the Madrid conurbation. Our objectives were to confirm the diagnosis, investigate the extent of symptoms and analyze the ecological factors contributing to ozone injury, particularly, the site water supply. Symptoms consisted of adaxial and intercostal stippling increasing with leaf age. Underlying stippling, cells in the upper mesophyll showed HR-like reactions typical of ozone stress. The surrounding cells showed further oxidative stress markers. These morphological and micromorphological markers of ozone stress were similar to those recorded in deciduous broadleaved species. However, stippling became obvious already at an AOT40 of 21 ppm•h and was primarily found at irrigated sites. Subsequent analyses showed that irrigated trees had their stomatal conductance increased and leaf life -span reduced whereas the leaf xeromorphy remained unchanged. These findings suggest a central role of water availability versus leaf xeromorphy for ozone symptom expression by cell injury in holm oak.

Application of integrated ozone biological aerated filters and membrane filtration in water reuse of textile effluents.

A combined process including integrated ozone-BAFs (ozone biological aerated filters) and membrane filtration was first applied for recycling textile effluents in a cotton textile mill with capacity of 5000 m(3)/d. Influent COD (chemical oxygen demand) in the range of 82-120 mg/L, BOD5 (5-day biochemical oxygen demand) of 12.6-23.1 mg/L, suspended solids (SSs) of 38-52 mg/L and color of 32-64° were observed during operation. Outflows with COD≤45 mg/L, BOD5≤7.6 mg/L, SS≤15 mg/L, color≤8° were obtained after being decontaminated by ozone-BAF with ozone dosage of 20-25 mg/L. Besides, the average removal rates of PVA (polyvinyl alcohol) and UV254 were 100% and 73.4% respectively. Permeate water produced by RO (reverse osmosis) could be reused in dyeing and finishing processes, while the RO concentrates could be discharged directly under local regulations with COD≤100 mg/L, BOD5≤21 mg/L, SS≤52 mg/L, color≤32°. Results showed that the combined process could guarantee water reuse with high quality, and solve the problem of RO concentrate disposal.

A chemically enhanced biological process for lowering operative costs and solid residues of industrial recalcitrant wastewater treatment.

An innovative process based on ozone-enhanced biological degradation, carried out in an aerobic granular biomass system (SBBGR--Sequencing Batch Biofilter Granular Reactor), was tested at pilot scale for tannery wastewater treatment chosen as representative of industrial recalcitrant wastewater. The results have shown that the process was able to meet the current discharge limits when the biologically treated wastewater was recirculated through an adjacent reactor where a specific ozone dose of 120 mg O3/L(influent) was used. The benefits produced by using ozone were appreciable even visually since the final effluent of the process looked like tap water. In comparison with the conventional treatment, the proposed process was able to reduce the sludge production by 25-30 times and to save 60% of operating costs. Molecular in situ detection methods were employed in combination with the traditional measurements (oxygen uptake rate, total protein content, extracellular polymeric substances and hydrophobicity) to evaluate microbial activity and composition, and the structure of the biomass. A stable presence of active bacterial populations was observed in the biomass with the simultaneous occurrence of distinctive functional microbial groups involved in carbon, nitrogen and sulphate removal under different reaction environments established within the large microbial aggregates. The structure and activity of the biomass were not affected by the use of ozone.

Determinants of ozone fluxes and metrics for ozone risk assessment in plants.

Tropospheric ozone concentration is increasing and represents a threat to single plants and whole ecosystems. The deleterious ozone effects mainly occur when (i) ozone concentration in the air builds up; (ii) the pollutant enters the leaf through stomatal uptake, and (iii) ozone-produced reactive oxygen species are not efficiently scavenged by leaf antioxidants and then oxidize leaf tissues. The sensitivity of plants to ozone is species-specific, and a correct risk assessment should be based on a metric that correctly takes into account the ambient concentration of ozone, the physiological control on stomatal apertures, and the efficiency of leaf antioxidant system. Current methodologies have been analysed to evaluate ozone risk assessment, and, by phasing-in and phasing out sources and sinks of ozone, elements of improvements for the current metrics have been suggested.

The impact of tropospheric ozone pollution on trial plot winter wheat yields in Great Britain - an econometric approach.

Numerous experiments have demonstrated reductions in the yields of cereal crops due to tropospheric O(3), with losses of up to 25%. However, the only British econometric study on O(3) impacts on winter wheat yields, found that a 10% increase in AOT40 would decrease yields by only 0.23%. An attempt is made here to reconcile these observations by developing AOT40 maps for Great Britain and matching levels with a large number of standardised trial plot wheat yields from many sites over a 13-year period. Panel estimates (repeated measures on the same plots with time) show a 0.54% decrease in yields and it is hypothesised that plant breeders may have inadvertently selected for O(3) tolerance in wheat. Some support for this is provided by fumigations of cultivars of differing introduction dates. A case is made for the use of econometric as well as experimental studies in prediction of air pollution induced crop loss.

Safety and quality assessment during the ozonation of cloudy apple juice.

Traditionally, ozone processing within the food industry has focused on solid foods by either gaseous treatment or washing with ozonized water. However, with the FDA's approval of ozone as a direct additive to food, the potential for liquid applications has emerged. This study investigates the effect of ozone processing on microbial inactivation (E. coli ATCC 25922 and NCTC 12900) and quality parameters (color, phenolic content) of cloudy apple juice. Apple juice samples were ozonated at room temperature (20 ± 1.5 °C) with a generated ozone concentration of 0.048 mg O(3) at a constant flow rate of 0.12 L/min and treatment time of 0 to 10 min. E. coli inactivation kinetics in apple juice were described quantitatively by using the Shoulder log-linear and the Weibull model. Ozone treatment of E. coli in apple juice demonstrate that a desired 5 log reduction can be achieved within 5 min. Apple juice color (L*, a*, and b*) and total phenols were significantly affected by ozone concentration and treatment time.

Moving toward effective ozone flux assessment.

We present a comment about "Ozone risk assessment for plants: central role of metabolism-dependent changes in reducing power" by Dizengremel, Le Thiec, Bagard, and Jolivet. As tools for summarizing plant O(3) sensitivity in simple indices, Dizengremel et al. suggest: reducing power, as antioxidant regeneration through the Halliwell/Asada cycle requires NADPH from the photosynthetic light reaction; Rubisco/PEPc ratio, as an index of the energy balance between anabolic and catabolic reactions; and water-use efficiency as a time-integrated approximation of the carbon gain to stomatal O(3) uptake ratio. The scientific background is solid, and simple enough (although expensive) to be translated into modelling and routine use. In the last decade, several approaches have been developed, mostly by using photosynthesis as a metric of defence. All these approaches should be experimentally tested in different and realistic conditions, before the results are transferred to the field and used in effective O(3) flux modelling and assessment.

Ozone risk assessment for plants: central role of metabolism-dependent changes in reducing power.

The combination of stomatal-dependent ozone flux and total ascorbate level is currently presented as a correct indicator for determining the degree of sensitivity of plants to ozone. However, the large changes in carbon metabolism could play a central role in the strategy of the foliar cells in response to chronic ozone exposure, participating in the supply of reducing power and carbon skeletons for repair and detoxification, and modifying the stomatal mode of functioning. To reinforce the accuracy of the definition of the threshold for ozone risk assessment, it is proposed to also consider the redox pool (NAD(P)H), the ratio between carboxylases and the water use efficiency as indicators of the differential ozone tolerance of plants.

A bench-scale, cost effective and simple method to elicit Lycopersicon esculentum cv. PKM1 (tomato) plants against Cucumber mosaic virus attack using ozone-mediated inactivated Cucumber mosaic virus inoculum.

Studies were undertaken to evaluate ozone for inactivation of Cucumber mosaic virus present in the inoculum and to stimulate Lycopersicon esculentum cv. PKM1 (tomato) plants against Cucumber mosaic virus infection by using the inactivated Cucumber mosaic virus inoculum. Application of a T(4) (0.4mg/l) concentration of ozone to the inoculum containing Cucumber mosaic virus resulted in complete inactivation of the virus. The inactivated viral inoculum was mixed with a penetrator (delivery agent), referred to as T(4) preparation, and it was evaluated for the development of systemic acquired resistance in the tomato plants. Application of a T(4) preparation 5 days before inoculation with the Cucumber mosaic virus protected tomato plants from the effects of Cucumber mosaic virus. Among the components of the inactivated virus tested, coat protein subunits and aggregates were responsible for the acquired resistance in tomato plants. In field trials, the results of enzyme-linked immunosorbent assay revealed that, Cucumber mosaic virus accumulation was significantly less for all the test plants (16%) sprayed with the T(4) preparation than untreated control plants (89.5%) at 28 days postinoculation (dpi). A remarkable increase in the activities of the total soluble phenolics (10-fold) and salicylic acid (16-fold) was detected 5 days after the treatment in foliar extracts of test plants relative to untreated control plants. The results showed that treatment of tomato plants with inactivated viral inoculum led to a significant enhancement of protection against Cucumber mosaic virus attack in a manner that mimics a real pathogen and induces systemic acquired resistance.

Uncertainty in prediction of disinfection performance.

Predicting the disinfection performance of a full-scale reactor in drinking water treatment is associated with considerable uncertainty. In view of quantitative risk analysis, this study assesses the uncertainty involved in predicting inactivation of Cryptosporidium parvum oocysts for an ozone reactor treating lake water. A micromodel is suggested which quantifies inactivation by stochastic sampling from density distributions of ozone exposure and lethal ozone dose. The ozone exposure distribution is computed with a tank in series model that is derived from tracer data and measurements of flow, ozone concentration and ozone decay. The distribution of lethal ozone doses is computed with a delayed Chick-Watson model which was calibrated by Sivaganesan and Marinas [2005. Development of a Ct equation taking into consideration the effect of Lot variability on the inactivation of Cryptosporidium parvum oocysts with ozone. Water Res. 39(11), 2429-2437] utilizing a large number of inactivation studies. Parameter uncertainty is propagated with Monte Carlo simulation and the probability of attaining given inactivation levels is assessed. Regional sensitivity analysis based on variance decomposition ranks the influence of parameters in determining the variance of the model result. The lethal dose model turns out to be responsible for over 90% of the output variance. The entire analysis is re-run for three exemplary scenarios to assess the robustness of the results in view of changing inputs, differing operational parameters or revised assumptions about the appropriate model. We argue that the suggested micromodel is a versatile approach for characterization of disinfection reactors. The scheme developed for uncertainty assessment is optimal for model diagnostics and effectively supports the management of uncertainty.

Synopsis of the CASIROZ case study: carbon sink strength of Fagus sylvatica L. in a changing environment--experimental risk assessment of mitigation by chronic ozone impact.

Databases are needed for the ozone (O(3)) risk assessment on adult forest trees under stand conditions, as mostly juvenile trees have been studied in chamber experiments. A synopsis is presented here from an integrated case study which was conducted on adult FAGUS SYLVATICA trees at a Central-European forest site. Employed was a novel free-air canopy O(3) fumigation methodology which ensured a whole-plant assessment of O(3) sensitivity of the about 30 m tall and 60 years old trees, comparing responses to an experimental 2 x ambient O(3) regime (2 x O(3), max. 150 nl O(3) l (-1)) with those to the unchanged 1 x ambient O(3) regime (1 x O(3)=control) prevailing at the site. Additional experimentation on individual branches and juvenile beech trees exposed within the forest canopy allowed for evaluating the representativeness of young-tree and branch-bag approaches relative to the O(3) sensitivity of the adult trees. The 2 x O(3) regime did not substantially weaken the carbon sink strength of the adult beech trees, given the absence of a statistically significant decline in annual stem growth; a 3 % reduction across five years was demonstrated, however, through modelling upon parameterization with the elaborated database. 2 x O(3) did induce a number of statistically significant tree responses at the cell and leaf level, although the O(3) responsiveness varied between years. Shade leaves displayed an O(3) sensitivity similar to that of sun leaves, while indirect belowground O(3) effects, apparently mediated through hormonal relationships, were reflected by stimulated fine-root and ectomycorrhizal development. Juvenile trees were not reliable surrogates of adult ones in view of O(3) risk assessment. Branch sections enclosed in (climatized) cuvettes, however, turned out to represent the O(3) sensitivity of entire tree crowns. Drought-induced stomatal closure decoupled O(3) intake from O(3) exposure, as in addition, also the "physiologically effective O(3) dose" was subject to change. No evidence emerged for a need to lower the "Critical Level for Ozone" in risk assessment of forest trees, although sensitive tree parameters did not necessarily reflect a linear relationship to O(3) stress. Exposure-based concepts tended to overestimate O(3) risk under drought, which is in support of current efforts to establish flux-related concepts of O(3) intake in risk assessment.

Modelling ozone effects on adult beech trees through simulation of defence, damage, and repair costs: Implementation of the CASIROZ ozone model in the ANAFORE forest model.

Ozone affects adult trees significantly, but effects on stem growth are hard to prove and difficult to correlate with the primary sites of ozone damage at the leaf level. To simulate ozone effects in a mechanistic way, at a level relevant to forest stand growth, we developed a simple ozone damage and repair model (CASIROZ model) that can be implemented into mechanistic photosynthesis and growth models. The model needs to be parameterized with cuvette measurements on net photosynthesis and dark respiration. As the CASIROZ ozone sub-model calculates effects of the ozone flux, a reliable representation of stomatal conductance and therefore ozone uptake is necessary to allow implementation of the ozone sub-model. In this case study the ozone sub-model was used in the ANAFORE forest model to simulate gas exchange, growth, and allocation. A preliminary run for adult beech (FAGUS SYLVATICA) under different ozone regimes at the Kranzberg forest site (Germany) was performed. The results indicate that the model is able to represent the measured effects of ozone adequately, and to distinguish between immediate and cumulative ozone effects. The results further help to understand ozone effects by distinguishing defence from damage and repair. Finally, the model can be used to extrapolate from the short-term results of the field study to long-term effects on tree growth. The preliminary simulations for the Kranzberg beech site show that, although ozone effects on yearly growth are variable and therefore insignificant when measured in the field, they could become significant at longer timescales (above 5 years, 5 % reduction in growth). The model offers a possible explanation for the discrepancy between the significant effects on photosynthesis (10 to 30 % reductions simulated), and the minor effects on growth. This appears to be the result of the strong competition and slow growth of the Kranzberg forest, and the importance of stored carbon for the adult beech (by buffering effects on carbon gain). We finally conclude that inclusion of ozone effects into current forest growth and yield models can be an important improvement into their overall performance, especially when simulating younger and less dense forests.