Continuing benefits of the Montreal Protocol and protection of the stratospheric ozone layer for human health and the environment.

The protection of Earth's stratospheric ozone (O3) is an ongoing process under the auspices of the universally ratified Montreal Protocol and its Amendments and adjustments. A critical part of this process is the assessment of the environmental issues related to changes in O3. The United Nations Environment Programme's Environmental Effects Assessment Panel provides annual scientific evaluations of some of the key issues arising in the recent collective knowledge base. This current update includes a comprehensive assessment of the incidence rates of skin cancer, cataract and other skin and eye diseases observed worldwide; the effects of UV radiation on tropospheric oxidants, and air and water quality; trends in breakdown products of fluorinated chemicals and recent information of their toxicity; and recent technological innovations of building materials for greater resistance to UV radiation. These issues span a wide range of topics, including both harmful and beneficial effects of exposure to UV radiation, and complex interactions with climate change. While the Montreal Protocol has succeeded in preventing large reductions in stratospheric O3, future changes may occur due to a number of natural and anthropogenic factors. Thus, frequent assessments of potential environmental impacts are essential to ensure that policies remain based on the best available scientific knowledge.

The response of aquatic ecosystems to the interactive effects of stratospheric ozone depletion, UV radiation, and climate change.

Variations in stratospheric ozone and changes in the aquatic environment by climate change and human activity are modifying the exposure of aquatic ecosystems to UV radiation. These shifts in exposure have consequences for the distributions of species, biogeochemical cycles, and services provided by aquatic ecosystems. This Quadrennial Assessment presents the latest knowledge on the multi-faceted interactions between the effects of UV irradiation and climate change, and other anthropogenic activities, and how these conditions are changing aquatic ecosystems. Climate change results in variations in the depth of mixing, the thickness of ice cover, the duration of ice-free conditions and inputs of dissolved organic matter, all of which can either increase or decrease exposure to UV radiation. Anthropogenic activities release oil, UV filters in sunscreens, and microplastics into the aquatic environment that are then modified by UV radiation, frequently amplifying adverse effects on aquatic organisms and their environments. The impacts of these changes in combination with factors such as warming and ocean acidification are considered for aquatic micro-organisms, macroalgae, plants, and animals (floating, swimming, and attached). Minimising the disruptive consequences of these effects on critical services provided by the world's rivers, lakes and oceans (freshwater supply, recreation, transport, and food security) will not only require continued adherence to the Montreal Protocol but also a wider inclusion of solar UV radiation and its effects in studies and/or models of aquatic ecosystems under conditions of the future global climate.

Environmental effects of stratospheric ozone depletion, UV radiation, and interactions with climate change: UNEP Environmental Effects Assessment Panel, Update 2021.

The Environmental Effects Assessment Panel of the Montreal Protocol under the United Nations Environment Programme evaluates effects on the environment and human health that arise from changes in the stratospheric ozone layer and concomitant variations in ultraviolet (UV) radiation at the Earth's surface. The current update is based on scientific advances that have accumulated since our last assessment (Photochem and Photobiol Sci 20(1):1-67, 2021). We also discuss how climate change affects stratospheric ozone depletion and ultraviolet radiation, and how stratospheric ozone depletion affects climate change. The resulting interlinking effects of stratospheric ozone depletion, UV radiation, and climate change are assessed in terms of air quality, carbon sinks, ecosystems, human health, and natural and synthetic materials. We further highlight potential impacts on the biosphere from extreme climate events that are occurring with increasing frequency as a consequence of climate change. These and other interactive effects are examined with respect to the benefits that the Montreal Protocol and its Amendments are providing to life on Earth by controlling the production of various substances that contribute to both stratospheric ozone depletion and climate change.

Environmental effects of stratospheric ozone depletion, UV radiation, and interactions with climate change: UNEP Environmental Effects Assessment Panel, Update 2020.

This assessment by the Environmental Effects Assessment Panel (EEAP) of the United Nations Environment Programme (UNEP) provides the latest scientific update since our most recent comprehensive assessment (Photochemical and Photobiological Sciences, 2019, 18, 595-828). The interactive effects between the stratospheric ozone layer, solar ultraviolet (UV) radiation, and climate change are presented within the framework of the Montreal Protocol and the United Nations Sustainable Development Goals. We address how these global environmental changes affect the atmosphere and air quality; human health; terrestrial and aquatic ecosystems; biogeochemical cycles; and materials used in outdoor construction, solar energy technologies, and fabrics. In many cases, there is a growing influence from changes in seasonality and extreme events due to climate change. Additionally, we assess the transmission and environmental effects of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is responsible for the COVID-19 pandemic, in the context of linkages with solar UV radiation and the Montreal Protocol.

Environmental effects of stratospheric ozone depletion, UV radiation and interactions with climate change: UNEP Environmental Effects Assessment Panel, update 2019.

This assessment, by the United Nations Environment Programme (UNEP) Environmental Effects Assessment Panel (EEAP), one of three Panels informing the Parties to the Montreal Protocol, provides an update, since our previous extensive assessment (Photochem. Photobiol. Sci., 2019, 18, 595-828), of recent findings of current and projected interactive environmental effects of ultraviolet (UV) radiation, stratospheric ozone, and climate change. These effects include those on human health, air quality, terrestrial and aquatic ecosystems, biogeochemical cycles, and materials used in construction and other services. The present update evaluates further evidence of the consequences of human activity on climate change that are altering the exposure of organisms and ecosystems to UV radiation. This in turn reveals the interactive effects of many climate change factors with UV radiation that have implications for the atmosphere, feedbacks, contaminant fate and transport, organismal responses, and many outdoor materials including plastics, wood, and fabrics. The universal ratification of the Montreal Protocol, signed by 197 countries, has led to the regulation and phase-out of chemicals that deplete the stratospheric ozone layer. Although this treaty has had unprecedented success in protecting the ozone layer, and hence all life on Earth from damaging UV radiation, it is also making a substantial contribution to reducing climate warming because many of the chemicals under this treaty are greenhouse gases.

Environmental effects of ozone depletion, UV radiation and interactions with climate change: UNEP Environmental Effects Assessment Panel, update 2017.

The Environmental Effects Assessment Panel (EEAP) is one of three Panels of experts that inform the Parties to the Montreal Protocol. The EEAP focuses on the effects of UV radiation on human health, terrestrial and aquatic ecosystems, air quality, and materials, as well as on the interactive effects of UV radiation and global climate change. When considering the effects of climate change, it has become clear that processes resulting in changes in stratospheric ozone are more complex than previously held. Because of the Montreal Protocol, there are now indications of the beginnings of a recovery of stratospheric ozone, although the time required to reach levels like those before the 1960s is still uncertain, particularly as the effects of stratospheric ozone on climate change and vice versa, are not yet fully understood. Some regions will likely receive enhanced levels of UV radiation, while other areas will likely experience a reduction in UV radiation as ozone- and climate-driven changes affect the amounts of UV radiation reaching the Earth's surface. Like the other Panels, the EEAP produces detailed Quadrennial Reports every four years; the most recent was published as a series of seven papers in 2015 (Photochem. Photobiol. Sci., 2015, 14, 1-184). In the years in between, the EEAP produces less detailed and shorter Update Reports of recent and relevant scientific findings. The most recent of these was for 2016 (Photochem. Photobiol. Sci., 2017, 16, 107-145). The present 2017 Update Report assesses some of the highlights and new insights about the interactive nature of the direct and indirect effects of UV radiation, atmospheric processes, and climate change. A full 2018 Quadrennial Assessment, will be made available in 2018/2019.

Fighting fish parasites with photodynamically active chlorophyllin.

Water-soluble chlorophyll (chlorophyllin) was used in a phototoxic reaction against a number of fish ectoparasites such as Ichtyobodo, Dactylogyrus, Trichodina, and Argulus. Chlorophyllin is applied to the water at concentrations of several micrograms per milliliter for a predefined incubation time, and afterwards, the parasites are exposed to simulated solar radiation. Application in the dark caused only little damage to the parasites; likewise, light exposure without the addition of the photosensitizer was ineffective. In Ichthyobodo, 2 µg/mL proved sufficient with subsequent simulated solar radiation to almost quantitatively kill the parasites, while in Dactylogyrus, a concentration of about 6 µg/mL was necessary. The LD50 value for this parasite was 1.02 µg/mL. Trichodina could be almost completely eliminated at 2 µg/mL. Only in the parasitic crustacean Argulus, no killing could be achieved by a photodynamic reaction using chlorophyllin. Chlorophyllin is non-toxic, biodegradable, and can be produced at low cost. Therefore, we propose that chlorophyllin (or other photodynamic substances) are a possible effective countermeasure against several ectoparasites in ponds and aquaculture since chemical remedies are either forbidden and/or ineffective.

Treatment of ichthyophthiriasis with photodynamically active chlorophyllin.

Water-soluble chlorophyll (chlorophyllin) exerts pronounced photodynamic activity on fish parasites. In order to determine its potential as a remedy against ectoparasites in fish carps were incubated in water with defined concentrations of chlorophyllin. The main focus of the experiments was on the ciliate Ichthyophthirius multifiliis (Fouquet) which is responsible for considerable losses in livestock in aquaculture. As malachite green, which in the past efficiently cured infected fishes, is banned because of its possible carcinogenicity; no effective remedy is presently available in aquaculture to treat ichthyophthiriasis. Using chlorophyllin, the number of trophonts was significantly reduced (more than 50 %) after 3 h incubation of infested fish at 2 and 4 mg/L and subsequent irradiation with simulated solar radiation. The lack of reinfection after light treatment indicates that also the remaining parasites have lost their multiplication capacity. In the controls (no chlorophyllin and no light, light but no chlorophyllin, or chlorophyllin but no light), no reduction of the I. multifiliis infection was observed. We propose that chlorophyllin (or other photodynamic substances) is a possible effective countermeasure against I. multifiliis and other ectoparasites in aquaculture.

Chlorophyllin for the control of Ichthyophthirius multifiliis (Fouquet).

Tests were conducted to determine the efficiency of the photodynamic substance chlorophyllin to kill different life stages of the protozoan parasite Ichthyophthirius mulftifiliis (Fouquet) which causes the white spot disease in many freshwater fish species. This study has developed a new treatment for the control of ichthyophthiriosis, and demonstrated that non-toxic water-soluble chlorophyll (chlorophyllin) is a potential remedial agent at low concentrations for this serious parasitic disease.

Effects of UV radiation on aquatic ecosystems and interactions with climate change.

The health of freshwater and marine ecosystems is critical to life on Earth. The impact of solar UV-B radiation is one potential stress factor that can have a negative impact on the health of certain species within these ecosystems. Although there is a paucity of data and information regarding the effect of UV-B radiation on total ecosystem structure and function, several recent studies have addressed the effects on various species within each trophic level. Climate change, acid deposition, and changes in other anthropogenic stressors such as pollutants alter UV exposure levels in inland and coastal marine waters. These factors potentially have important consequences for a variety of aquatic organisms including waterborne human pathogens. Recent results have demonstrated the negative impacts of exposure to UV-B radiation on primary producers, including effects on cyanobacteria, phytoplankton, macroalgae and aquatic plants. UV-B radiation is an environmental stressor for many aquatic consumers, including zooplankton, crustaceans, amphibians, fish, and corals. Many aquatic producers and consumers rely on avoidance strategies, repair mechanisms and the synthesis of UV-absorbing substances for protection. However, there has been relatively little information generated regarding the impact of solar UV-B radiation on species composition within natural ecosystems or on the interaction of organisms between trophic levels within those ecosystems. There remains the question as to whether a decrease in population size of the more sensitive primary producers would be compensated for by an increase in the population size of more tolerant species, and therefore whether there would be a net negative impact on the absorption of atmospheric carbon dioxide by these ecosystems. Another question is whether there would be a significant impact on the quantity and quality of nutrients cycling through the food web, including the generation of food proteins for humans. Interactive effects of UV radiation with changes in other stressors, including climate change and pollutants, are likely to be particularly important.

Phototaxis and photophobic responses in Stentor coeruleus. Action spectrum and role of Ca2+ fluxes.

Negative phototactic orientation, step-up photophobic responses and light-induced action potentials have been studied in the ciliate Stentor coeruleus. A resolved action spectrum, based on fluence rate-response curves, is consistent with stentorin as the photoreceptor. Calcium flux blockers prolong the response time for ciliary stop and reversal and inhibit step-up photophobic responses. Drugs believed to affect the membrane-bound calcium pump likewise inhibit phobic responses. On the other hand, alpha-phosphatidic acid promotes Ca2+-influx and enhances the photophobic sensitivity of the organism, thus providing an unambiguous evidence for the role of Ca2+ influx. A change in the response time decreases the degree of phototactic orientation, indicating that negative phototaxis in this organism is brought about by subsequent phobic responses of individual rows of cilia as the associated photoreceptor granules experience an increase in light intensity when the organism rotates during forward locomotion in lateral light.

Overview of the Liulin type instruments for space radiation measurement and their scientific results.

Ionizing radiation is recognized to be one of the main health concerns for humans in the space radiation environment. Estimation of space radiation effects on health requires the accurate knowledge of the accumulated absorbed dose, which depends on the global space radiation distribution, solar cycle and local shielding generated by the 3D mass distribution of the space vehicle. This paper presents an overview of the spectrometer-dosimeters of the Liulin type, which were developed in the late 1980s and have been in use since then. Two major measurement systems have been developed by our team. The first one is based on one silicon detector and is known as a Liulin-type deposited energy spectrometer (DES) (Dachev et al., 2002, 2003), while the second one is a dosimetric telescope (DT) with two or three silicon detectors. The Liulin-type instruments were calibrated using a number of radioactive sources and particle accelerators. The main results of the calibrations are presented in the paper. In the last section of the paper some of the most significant scientific results obtained in space and on aircraft, balloon and rocket flights since 1989 are presented.

A rapid, simple method for the isolation and characterization of the photoreceptor of Dictyostelium discoideum amoebae.

A membrane-bound 45.5 kDa protein has been isolated from Dictyostelium discoideum amoebae. It shows an absorption spectrum, which closely resembles the action spectrum for amoebal phototaxis, leading to the conclusion that this protein might play an important role in the photoreception of Dictyostelium amoebae. For further characterization we employed phase partition in an aqueous polymer two-phase system, which was developed by Widell and Larsson for the separation of plasma membrane proteins of higher plants. This method clearly shows that the 45.5 kDa protein is a plasma membrane protein and not an intracellular protein. Furthermore, by using phase systems with increasing polymer concentrations, this simple and rapid purification of plasma membrane proteins allowed us to isolate the putative photoreceptor in one single step. Compared to standard biochemical methods phase partition provides an enormous facilitation of the isolation of D. discoideum membrane proteins.

Isolation and characterisation of chemotactic mutants of Chlamydomonas reinhardtii obtained by insertional mutagenesis.

The swimming behaviour of the green flagellated protist Chlamydomonas reinhardtii is influenced by several different external stimuli including light and chemical attractants. Common components are involved in both the photo- and chemo-sensory transduction pathways, although the nature and organisation of these pathways are poorly understood. To learn more about the mechanism of chemotaxis in Chlamydomonas, we have generated nonchemotactic strains by insertional mutagenesis. The arginine-requiring strain arg7-8 was transformed with DNA carrying the wild-type ARG7 gene. Of the 8630 arginine-independenttransformants obtained, five are defective in their chemotaxis towards various sugars. Two of the mutants (CTX2 and CTX3) are blocked only in their response to xylose. Mutant CTX1 is blocked in its response to xylose, maltose and mannitol, but displays normal taxis to sucrose. Mutants CTX4 and CTX5 lack chemotactic responses to all sugars tested. CTX1, CTX4 and CTX5 represent novel chemotactic phenotypes not previously obtained using ultra-violet or chemical mutagenesis. Genetic analysis confirms that each mutation maps to a single nuclear locus that is unlinked to the mating-type locus. Further analysis of CTX4 indicates that the mutant allele is tagged by the transforming ARG7 DNA. CTX4 appears to be defective in a component specific for chemotactic signal transduction since it exhibits wild-type photobehavioural responses (phototaxis and photoshock) as well as the wild-type responses of EGTA-induced trans-flagellum inactivation and acid-induced deflagellation. Insertional mutagenesis has thus permitted the generation of novel chemotactic mutants that will be of value in the molecular dissection of the signalling machinery.

Graviperception in the flagellate Euglena gracilis during a shuttle space flight.

During a recent space flight, gravitaxis of the unicellular photosynthetic flagellate, Euglena gracilis, was studied on board of the American shuttle Columbia. Accelerations were varied between 0 and 1.5 x g using a slow rotating centrifuge microscope (NIZEMI). The cells showed a sigmoidal response curve for the dependence of the precision of gravitaxis on acceleration which is indicative of the involvement of an active, physiological gravireceptor with a threshold at g-values < or = 0.16 x g and a saturation at g-values > or = 1 x g. No adaptation to microgravity was found during the prolonged space mission. After return the cells showed a normal gravitactic behavior at 1 x g. Since the cells are heavier than water, their swimming velocity is affected by sedimentation. The velocity distribution at different accelerations closely follows Stokes' law for sedimentation indicating that, in contrast to the ciliate Paramecium, E. gracilis, does not show any gravikinesis.

Influence of accelerations on the spatial orientation of Loxodes and Paramecium.

The gravitactic ciliates Paramecium and Loxodes were cultivated for 15 days in space during the IML-2 spacelab mission. At dedicated times their behavioral responses to different accelerations between 10(-3) x g and 1.5 x g were investigated by using a slow rotating centrifuge microscope (NIZEMI). The threshold for gravitaxis of Paramecium was found to be at > 0.16 x g and < or = 0.3 x g. No adaptation of Paramecium to the conditions of weightlessness was observed over the duration of 15 days. Loxodes showed no graviresponses to increasing accelerations, though it demonstrated gravitaxis after return to earth.

Signal perception and amplification in photoresponses of cyanobacteria.

In addition to a number of other external stimuli, the gliding cyanobacterium, Phormidium uncinatum, uses light as a clue to control movement in order to find and stay in a microhabitat with optimal conditions for growth and survival of the population. Of the three photoresponses developed in cyanobacteria, phototaxis, photokinesis and photophobic responses, the latter are the most prominent for this task. Step-down phobic responses are mediated by the photosynthetic pigments and are linked to the linear electron-transport chain. A change in the fluence rate is reflected by a change in the proton-motive force which is amplifi.

A novel express bioassay for detecting toxic substances in water by recording rhodopsin-mediated photoelectric responses in Chlamydomonas cell suspensions.

The influence of Cu2+, Zn2+, Cd2+, Pb2+ and formaldehyde on rhodopsin-mediated photoelectric responses in the green flagellate Chlamydomonas reinhardtii was investigated using three modifications of a recently developed population method for electrical recording (in nonoriented, phototactically preoriented (PO) and gravitactically preoriented cell suspensions). The addition of the heavy metal ions at concentrations several times lower than those known to affect swimming velocity and other physiological parameters in photosynthetic flagellates led to a rapid (one to several minutes) inhibition of the responses. Formaldehyde induced a significant temporary increase in the gravi-orientation of the cells simultaneously with an inhibition of their photoelectric cascade, photo-orientation and motility. The signals recorded in PO suspensions were more sensitive to all tested toxic substances than those recorded from nonoriented cells and indicated a switch from negative to positive phototaxis in the presence of the toxic substances. Of the two major components of the photoelectric cascade, the regenerative response was more sensitive to the tested heavy metal ions, but not to formaldehyde, than the photoreceptor current. The results obtained show that measurement of the photoinduced electrical responses in Chlamydomonas cell suspensions is a powerful novel bioassay for testing environmental pollutants in water samples.

High-energy carbon ion irradiation and the inhibition of negative gravitaxis in Euglena gracilis Z.

PURPOSE: To study the effects of high-energy carbon ion irradiation on negative gravitaxis in the photosynthetic flagellate Euglena gracilis strain Z in a dose-response-dependent manner. MATERIALS AND METHODS: Cells were exposed to 290 MeV amu(-1) carbon ion from the Heavy Ion Medical Accelerator in Chiba (HIMAC) at 12 doses (0, 1, 5, 7.5, 10, 15, 20, 50, 75, 100, 150, 200 Gy for water). r was used to quantify negative gravitaxis observed in a recently developed biomonitoring system. RESULTS: Negative gravitaxis of Euglena was significantly affected by irradiation at a dose >7.5 Gy, 28 h after irradiation. Negative gravitaxis recovered after a few days. The rising phase of r during the first 30 s of orientation was independent of irradiation dose. The recovery mechanism itself was damaged by 290 MeV amu(-1) carbon ions at a dose >50 Gy. CONCLUSIONS: The results indicate that negative gravitaxis in Euglena may be affected by the radiation experienced during a space experiment especially during the occurrence of solar flares. The analyses of time needed to reorient to the gravitational vector suggest that the steering control of the flagellar apparatus is affected by exposure to the 290 MeV amu(-1) carbon ion irradiation, resulting in an altered beating pattern of a flagellum and/or changes of the cell form during reorientation.

Interactive image analysis system to determine the motility and velocity of cyanobacterial filaments.

An interactive image analysis system has been developed to analyse and quantify the percentage of motile filaments and the individual linear velocities of organisms. The technique is based on the "difference" image between two digitized images taken from a time-lapse video recording 80 s apart which is overlaid on the first image. The bright lines in the difference image represent the paths along which the filaments have moved and are measured using a crosshair cursor controlled by the mouse. Even short exposure to solar ultraviolet radiation strongly impairs the motility of the gliding cyanobacterium Phormidium uncinatum, while its velocity is not likewise affected. These effects are not due to either type I (free radical formation) or type II (singlet oxygen production) photodynamic reactions, since specific quenchers and scavengers, indicative of these reactions, failed to be effective.