Antioxidant metabolism in Xenopus laevis embryos is affected by stratospheric balloon flight

To test the effects of low levels of radiation from space on living organisms, we flew Xenopus laevis embryos at different stages of development on a stratospheric balloon (BI.R.BA mission). After recovery, different parameters were analyzed to assess the effects of flight, with particular regard to oxidative stress damage. Because of failed temperature control during flight, the flight shielded embryos (FC) could not be used for biochemical or morphological comparisons. In contrast, the incubation conditions (i.e. temperature, containers, volumes) for the flight embryos (F) were parallel to those for the ground controls. Mortality data show that younger embryos (16 h) flown on the balloon (F) are more sensitive to radiation exposure than older ones (40 h and 6 days). Exposure during flight lowered the antioxidant potential in all embryos, particularly older ones. These preliminary data demonstrate that flight on a stratospheric balloon might affect antioxidant metabolism, though it is not yet possible to correlate these results with low radiation exposure during flight.     

Assessing invasion risk across taxa and habitats: life stage as a determinant of invasion success

Aim Many aquatic invertebrates produce dormant life‐history stages as a means to endure inhospitable environments and to facilitate natural long‐distance dispersal, yet we have little understanding of the role of dormant stages as a mechanism for human‐mediated introductions of non‐indigenous species. We explore the survival of invertebrate dormant eggs in collected ships’ ballast sediment over a 1‐year period to determine relative invasion potential across taxa (i.e. rotifers, copepods, cladocerans and bryozoans) and different habitats (freshwater, marine). Location Canadian Atlantic and Pacific coasts and Laurentian Great Lakes. Methods During 2007 and 2008, 19 ballast samples were collected as a part of a larger study. The degradation rate of dormant eggs was assessed by enumerating dormant eggs and by conducting viability hatching experiments. Results Taxa examined included rotifers, copepods, anomopods, onychopods and bryozoans. Dormant eggs of rotifers degraded at the highest rate of all taxa examined, with no viable eggs remaining within 10 months. Copepods showed a less rapid degradation rate than rotifers. The degradation rate of anomopod dormant eggs was significantly slower than that of both rotifers and copepods. Onychopods and bryozoans did not visibly degrade at all over 12 months. Viability hatching experiments were successful for rotifers, copepods, and anomopods. Onychopods and bryozoans did not hatch during any of the three hatching trials. Main conclusions Dormancy is not equally beneficial to all invertebrate taxa. Our results indicate that dormant eggs of rotifers and copepods degrade at a rapid rate and may not pose high invasion risk. In contrast, the slow degradation rate of anomopod dormant eggs and the lack of degradation of onychopod and bryozoan dormant eggs could result in high invasion risk because of their accumulation in ballast tanks. Species having resistant dormant eggs mostly belong to freshwater taxa making freshwater habitats at higher invasion risk by dormant invertebrates than marine habitats.     

Macrotrachela quadricornifera featured in a space experiment

Macrotrachela quadricornifera, a bdelloid rotifer, is the animal model of an experiment scheduled on the International Space Station (ISS). The focus of the experiment is the role of the cytoskeleton during oogenesis and early development. Bdelloids will fly desiccated, be rehydrated on the ISS and cultivated for five generations. We present the outline of the ISS experiment, the expectations and the state-of-the-art of ground-based research run to date on the major topics of the planned experiment: anhydrobiosis and embryogenesis. Anhydrobiosis focuses on two major aspects, storage conditions that enhance recovery rate and comparison of the resistance between dormant and active rotifers to UV radiation and other environmental injuries. Embryogenesis has been approached at the morphological level under ground conditions: developing embryos have been studied by light and confocal microscopes.     

Amphora margalefii Tomàs var. lacustris P. Sánchez var. nova,a new brackish water diatom

The rotifer Brachionus plicatilis can grow in a wide range of salinities and temperatures, but rapid shifts in both salinity and temperature may result in immobilized, non-swimming rotifers. The goal of this study was to examine the effect of perturbations in temperature and salinity on the swimming pattern of the rotifer.Only slight changes in mobility were observed when rotifers were exposed to changes in temperature (from 20 °C to 8–30 °C) and to an increase in salinity (from 20% to 30%). When the salinity was reduced to 15% and 5%, the proportion of mobile rotifers was reduced to 50% and 5%, respectively. The rotifers were throughout more resistant to perturbations in temperature than to those of salinity.Combined temperature and salinity perturbations compared to perturbations in each factor separately suggested a synergetic effect of temperature and salinity on the rotifers locomotion. Transfer from cultivation conditions to low salinity (5%) and high temperature (28 °C) resulted in very low percent of mobile rotifers (0–10%). However, if the temperature was reduced to 8 °C concomitant with the changes in salinity, the percent of mobile rotifers was 85%.Rotifers use a high share of their metabolic energy for locomotion, and it is therefore not surprising that perturbations in salinity and temperature may result in partial or complete immobilization.     

Morphological and physiological responses of bean plants to supplemental UV radiation in a Mediterranean climate

During the last few decades many experiments have been performed to evaluate the responses of plants to enhanced solar UV-B radiation (280–320 nm) that may occur because of stratospheric ozone depletion; most of them were performed in controlled environment conditions where plants were exposed to low photosynthetically active radiation (PAR) levels and high UV-B irradiance. Since environmental radiative regimes can play a role in the response of plants to UV-B enhancement, it appears doubtful whether it is valid to extrapolate the results from these experiments to plants grown in natural conditions. The objective of this work was to evaluate the effects on physiology and morphology of a bean (Phaseolus vulgaris L.) cultivar Nano Bobis, exposed to supplemental UV radiation in the open-air. UV-B radiation was supplied by fluorescent lamps to simulate a 20% stratospheric ozone reduction. Three groups of plants were grown: control (no supplemental UV), UV-A treatment (supplementation in the UV-A band) and UV-B treatment (supplemental UV-B and UV-A radiation). Each group was replicated three times. After 33 days of treatment plants grown under UV-B treatment had lower biomass, leaf area and reduced leaf elongation compared to UV-A treatment. No significant differences were detected in photosynthetic parameters, photosynthetic pigments and UV-B absorbing compounds among the three groups of plants. However, plants exposed to UV-A treatment showed a sort of 'stimulation' of their growth when compared to the control. The results of this experiment showed that plants may be sensitive to UV-A radiation, thus it is difficult to evaluate the specific effects of UV-B (280–320 nm) radiation from fluorescent lamps and it is important to choose the appropriate control. Environmental conditions strongly affect plant response to UV radiation so further field studies are necessary to assess the interaction between UV-B exposure and meteorological variability.     

Behavior and reproduction of invertebrate animals during and after a long-term microgravity: space experiments using an Autonomous Biological System (ABS).

Aquatic invertebrate animals such as Amphipods, Gastropods (pond snails), Ostracods and Daphnia (water flea) were placed in water-filled cylindrical vessels together with water plant (hornwort). The vessels were sealed completely and illuminated with a fluorescent lamp to activate the photosynthesis of the plant for providing oxygen within the vessels. Such ecosystem vessels, specially termed as Autonomous Biological System or ABS units, were exposed to microgravity conditions, and the behavior of the animals and their reproduction capacity were studied. Three space experiments were carried out. The first experiment used a Space shuttle only and it was a 10-day flight. The other two space experiments were carried out in the Space station Mir (Shuttle/Mir mission), and the flight units had been kept in microgravity for 4 months. Daphnia produced their offspring during a 10-day Shuttle flight. In the first Mir experiment, no Daphnia were detected when recovered to the ground. However, they were alive in the second Mir experiment. Daphnia were the most fragile species among the invertebrate animals employed in the present experiments. All the animals, i.e., Amphipods, pond snails, Ostracods and Daphnia had survived for 4 months in space, i.e., they had produced their offspring or repeated their life-cycles under microgravity. For the two Mir experiments, in both the flight and ground control ecosystem units, an inverse relationship was noted between the number of Amphipods and pond snails in each unit. Amphipods at 10 hours after the recovery to the ground frequently exhibited a movement of dropping straight-downward to the bottom of the units. Several Amphipods had their legs bent abnormally, which probably resulted from some physiological alterations during their embryonic development under microgravity. From the analysis of the video tape recorded in space, for Ostracods and Daphnia, a half of their population were looping under microgravity. Such looping animals could be observed still at the end of the 4 month stay in space. No looping behavior was noted for Amphipods and pond snails.     

Teratogenic, biochemical, and histological studies with mice prenatally exposed to 2.45-GHz microwave radiation

Pregnant CD-1 mice were exposed to 2.45-GHz continuous wave microwave radiation at an incident power density of 30 mW/cm2. The local specific absorption rate near the uterine area (deep colonic location), as determined from time-temperature profiles measured with a Vitek thermistor probe, was 40.2 mW/g. Groups of mice were exposed 8 hr per day through Days 1-6 or 6-15 of pregnancy. Other groups of animals were exposed to an elevated ambient temperature of 31 degrees C which increased the colonic temperature 2.3 degrees C, the same as that produced by the microwaves. Sham-irradiated groups of animals were treated exactly the same as the microwave-exposed animals. For the two conditions, temperature exposed and sham exposed, two groups of animals were used. One group was handled in the same manner as the microwave-irradiated group and the other group was not handled so as to evaluate the effects of stressing the animals by handling. Eleven groups of animals were used in the complete study: five groups for gestational Days 1-6, five groups for gestational Days 6-15, and one group of cage control animals. On Day 18 of gestation the dams of all experimental groups were sacrificed and their reproductive status was determined. The fetuses were examined for visceral and skeletal alterations. Brain cholinesterase activity and histology were evaluated in the groups exposed on Days 6-15. The results show that microwave radiation increases embryo lethality at the early stages of gestation (exposure Days 1-6). Fetal toxicity and teratogenicity were not significantly increased by exposure to microwaves on either Days 1-6 or 6-15 of gestation. Cholinesterase activity and histology of the brain of 18-day-old fetuses were not adversely affected.     

Effects of Ultraviolet Radiation on Locomotion and Orientation in Roughskin Newts (Taricha granulosa)

Environmental changes, including those associated with the atmosphere may significantly affect individual animals and ultimately populations. Ultraviolet (UV) radiation, perhaps increasing due to stratospheric ozone depletion, has been linked to mortality in a number of organisms, including amphibians. The eggs and larvae of certain amphibian species hatch at significantly lower rates when exposed to ambient ultraviolet light. Yet little is known about the sublethal effects of UV radiation. For example, UV radiation may affect specific behaviors of an animal that could alter its ability to survive. To examine if UV radiation affects amphibian behavior, we used roughskin newts ( Taricha granulosa ) as a model. Newts were exposed to low-level doses of UV in the laboratory and then tested in the field to examine if UV-exposed and control (no UV) newts differed in orientation towards water or in locomotor activity levels. UV-exposed and control newts both exhibited a significant orientation towards water in field tests but there was no significant difference in orientation between treatments. However, UV-exposed newts were significantly more active than control newts. Our results suggest that exposure to short-term low levels of UV radiation alters certain behaviors. Environmentally induced changes in behavior may have significant ecological and evolutionary consequences.     

Artificial and solar UV radiation induces strand breaks and cyclobutane pyrimidine dimers in Bacillus subtilis spore DNA

The loss of stratospheric ozone and the accompanying increase in solar UV flux have led to concerns regarding decreases in global microbial productivity. Central to understanding this process is determining the types and amounts of DNA damage in microbes caused by solar UV irradiation. While UV irradiation of dormant Bacillus subtilis endospores results mainly in formation of the "spore photoproduct" 5-thyminyl-5,6-dihydrothymine, genetic evidence indicates that an additional DNA photoproduct(s) may be formed in spores exposed to solar UV-B and UV-A radiation (Y. Xue and W. L. Nicholson, Appl. Environ. Microbiol. 62:2221-2227, 1996). We examined the occurrence of double-strand breaks, single-strand breaks, cyclobutane pyrimidine dimers, and apurinic-apyrimidinic sites in spore DNA under several UV irradiation conditions by using enzymatic probes and neutral or alkaline agarose gel electrophoresis. DNA from spores irradiated with artificial 254-nm UV-C radiation accumulated single-strand breaks, double-strand breaks, and cyclobutane pyrimidine dimers, while DNA from spores exposed to artificial UV-B radiation (wavelengths, 290 to 310 nm) accumulated only cyclobutane pyrimidine dimers. DNA from spores exposed to full-spectrum sunlight (UV-B and UV-A radiation) accumulated single-strand breaks, double-strand breaks, and cyclobutane pyrimidine dimers, whereas DNA from spores exposed to sunlight from which the UV-B component had been removed with a filter ("UV-A sunlight") accumulated only single-strand breaks and double-strand breaks. Apurinic-apyrimidinic sites were not detected in spore DNA under any of the irradiation conditions used. Our data indicate that there is a complex spectrum of UV photoproducts in DNA of bacterial spores exposed to solar UV irradiation in the environment.     

Are Freshwater Mixotrophic Ciliates Less Sensitive to Solar Ultraviolet Radiation than Heterotrophic Ones?1

We tested whether mixotrophic ciliates are more resistant to solar ultraviolet radiation (UVR) than heterotrophic ones because symbiotic algae can provide self-shading by cell matter absorption and eventually by direct UV screening from mycosporine-like amino acids (MAAs). Sensitivity of a natural assemblage to solar radiation was tested in experiments in the original lake and in a more UV transparent alpine lake after transplantation of the ciliates. In both lakes, the assemblage was exposed either to full sunlight, to photosynthetically active radiation only, or kept in the dark. In each lake, exposure was for 5 h at the surface and at the depth corresponding to the 10% attenuation depth at 320 nm. Overall, when the assemblage was exposed to surface UVR, only one out of four dominant mixotrophic ciliates, Vorticella chlorellata, was more resistant than heterotrophic species. The higher UV resistance in V. chlorellata was related to the presence of MAAs and the high percentage of ciliate volume occupied by algal symbionts. Our results indicate that effects of UVR were species-specific and depended on efficient screening of these wavelengths, but also on the depth preference of the ciliates and thus, on their previous exposure history to UVR.     

Mutation frequency of Dictyostelium discoideum spores exposed to the space environment.

Two strains of cellular slime mold Dictyostelium discoideum, a radiosensitive mutant and the parental wild-type strain, were used to investigate the effects of cosmic radiation on viability and mutation frequency at the spore stage for about 9 days in Space Shuttle of NASA. We measured little effect of space environment on viability and cell growth in the both strains as compared to ground controls. The mutation frequency of the flown spores were similar to that of ground control. These results suggest that there could be no effect of cosmic radiation, containing high linear energy transfer radiation at about 0.9 mSv/day as detected by real-time radiation monitoring device on the induction of mutation at the spore stage.     

Stress during dormancy: effect on recovery rates and life-history traits of anhydrobiotic animals

To test the influence of storage conditions on recovery and life-history traits of dormant animals we subjected cohorts of a bdelloid rotifer (Macrotrachela quadricornifera) and a nematode (Panagrolaimus rigidus) to three different conditions during a 26-d anhydrobiosis period. These conditions were (1) standard conditions in our laboratory (AC), (2) a flight to the U.S.A. and storage there (KSC), and (3) a flight to the U.S.A. followed by a flight on a Shuttle-to-Mir mission (Shuttle). Another cohort (HC) was maintained under hydrated conditions and served as reference for life-history traits of the desiccated samples. The recovery after dormancy and the residual life-history traits of the different cohorts were compared. Results indicate that (a) during dormancy the animals were indifferent to the test conditions, as evidenced from both recovery capacity and life history performances, (b) the anhydrobiotic period affected the experimental species differently. At recovery both animals resumed their life cycle, but while the rotifer appeared indifferent to the time spent in desiccation, the nematode modified its life history according to the duration of anhydrobiosis. Third, (c) induction of anhydrobiosis can act as a switching mechanism for animals capable of desiccation. Anhydrobiosis probably represents a suitable method to preserve organisms during stressful conditions allowing them to return to a normal life when convenient.     

Artificial and Solar UV Radiation Induces Strand Breaks and Cyclobutane Pyrimidine Dimers in Bacillus subtilis Spore DNA

The loss of stratospheric ozone and the accompanying increase in solar UV flux have led to concerns regarding decreases in global microbial productivity. Central to understanding this process is determining the types and amounts of DNA damage in microbes caused by solar UV irradiation. While UV irradiation of dormant Bacillus subtilis endospores results mainly in formation of the “spore photoproduct” 5-thyminyl-5,6-dihydrothymine, genetic evidence indicates that an additional DNA photoproduct(s) may be formed in spores exposed to solar UV-B and UV-A radiation (Y. Xue and W. L. Nicholson, Appl. Environ. Microbiol. 62:2221–2227, 1996). We examined the occurrence of double-strand breaks, single-strand breaks, cyclobutane pyrimidine dimers, and apurinic-apyrimidinic sites in spore DNA under several UV irradiation conditions by using enzymatic probes and neutral or alkaline agarose gel electrophoresis. DNA from spores irradiated with artificial 254-nm UV-C radiation accumulated single-strand breaks, double-strand breaks, and cyclobutane pyrimidine dimers, while DNA from spores exposed to artificial UV-B radiation (wavelengths, 290 to 310 nm) accumulated only cyclobutane pyrimidine dimers. DNA from spores exposed to full-spectrum sunlight (UV-B and UV-A radiation) accumulated single-strand breaks, double-strand breaks, and cyclobutane pyrimidine dimers, whereas DNA from spores exposed to sunlight from which the UV-B component had been removed with a filter (“UV-A sunlight”) accumulated only single-strand breaks and double-strand breaks. Apurinic-apyrimidinic sites were not detected in spore DNA under any of the irradiation conditions used. Our data indicate that there is a complex spectrum of UV photoproducts in DNA of bacterial spores exposed to solar UV irradiation in the environment.     

Ecological,Dynamic and Taxonomic Problems Due to <Emphasis Type="Italic">Ludwigia</Emphasis> (Onagraceae) in France

We analyzed meta-community structure of bdelloid rotifers colonizing mosses along an 80 meter section of Rio Valnava in NW Italy. Bdelloid rotifers are small animals living associated with a substratum; colonization in bdelloids can be produced by active animals moving along the riverbed, or by passive dormant propagules, moved by wind. To detect which kind of colonization might be stronger at different spatial scales, we designed a spatially nested sampling experiment at three hierarchical levels: (1) single sample, (2) 10 communities inside each pool, (3) complete section of 10 pools. Assessing species richness and species similarity of communities, and coherence and nestedness of bdelloid meta-communities, we found that different forces may drive species composition at different spatial scales: at the largest scale, colonization of propagules may over-ride direct dispersal between pools, while at the scale of the single pool, differential movements of species give a nested structure to the meta-communities. The number of species increased as the level of analysis increased, even though this study was carried out along only a small stream section.     

Experimental study of the embryotropic action of tetracycline]

The general toxic and embryotropic effect of tetracycline was studied on unbred pregnant albino rats. The antibiotic was used in concentrations of 1 and 0.1 mg/m3, the maximum permissible concentration (MPC). The animals were exposed to tetracycline inhalation for 20 days of their pregnancy. On the basis of the results it was concluded that tetracycline in the concentration at the level of the MPC, i.e. 0.1 mg/m3 had under the experimental conditions a negative effect on the pregnant animals. In concentrations of 1 and 0.1 mg/m3 it had in addition an embryotropic effect. It was also shown that the pregnant animals were more sensitive to the antibiotic than the non-pregnant ones.     

Thermoregulation as a limit to habitat use in alpine marmots (Marmota marmota)

Summary The body temperature (T _b) of free-living alpine marmots rose with activity; the higher the effective environmental temperature (T _e), the higher the rise. Maximum T _bof 40° C was reached at the time of greatest activity in late afternoon or evening. The activity pattern was strongly influenced by the microclimate. Up to an T _eof 25° C the animals spent more time above ground and were more active the higher T _ewas, but above 25° C this trend was reversed, and the animals withdrew increasingly into their burrows. On warm days the activity pattern was therefore bimodal and above ground presence was reduced, in contrast to cool days. Hence behavioural thermoregulation limits the available time for above ground activity on days with high T _ein this strictly diurnal species. We suggest that the alpine marmots' preference for south oriented slopes is due to the better conditions for hibernation there, the microclimate during summer is more favourable on northerly slopes. Thermoregulatory constraints could also keep alpine marmots away from lower elevations.     

Prediction of the changes in thermodynamic stability of proteins caused by single amino acid substitutions

The functional (synthetic) activity of blood lymphocytes and bone marrow hematopoietic cells in ground squirrels was studied in different seasons and at different stages of the torpor-arousal cycle. The effect of γ-irradiation on animals in different physiological states was also studied. The synthetic activity of cells was estimated from the amount of active RNA per unit DNA in the cell (parameter α). The α values in lymphocytes were minimal in hibernating animals (January–March), reached a peak upon their complete awakening (April), slightly decreased in the summer activity period, and decreased further in the prehibernation autumn period (November). During winter arousals between torpor bouts, this parameter reached the same values as in summer. The dynamics of parameter α in bone marrow hematopoietic cells were generally similar: minimal values in November and higher between torpor bouts than in summer. The peak of synthetic activity of proliferating hematopoietic cells recorded upon awakening from hibernation in April was mainly due to the accumulation of cells in the G₁ and G₂ phases of the cell cycle, and its decrease in summer reflected prevalent transition from G₂ to mitosis and then partly to G₀. In the torpor-arousal-euthermia cycle, two stages of awakening were distinguished, differing considerably in most of the test parameters. The synthetic activity and the total number of blood and bone marrow cells in ground squirrels irradiated in the state of torpor did not differ significantly from those in nonirradiated torpid animals. The adverse effect of radiation in animals irradiated at the initial stage of awakening was lesser than in animals irradiated in the active state, whereas animals at the second stage of awakening proved more vulnerable to acute irradiation. The physiological state of ground squirrels exposed to ionizing radiation at different phases of the torpor-arousal-euthermia cycle plays a key role in the dynamics of qualitative and quantitative characteristics of blood system cells. The results of this study indicate that the hypometabolic state of ground squirrels during hibernation is a factor of protection from the impact of ionizing radiation on the whole body and on the immune system in particular.     

Effects of UV-B radiation on growth and motility of four phytoplankton species

A depletion of the stratospheric ozone layer would result in an increased UV-B radiation, which could have harmful effects on marine organisms. The aim of this study was to determine the effects of an enhanced UV-B radiation (280–320 nin) on the motility and growth in four Swedish phytoplanklon species. The different plankton species were exposed to different doses of UV-B radiation during growth. The growth of the motile dinoflagellates, Gyronidium aureolum Hulburt (Ba 6), and Prorocentrum minimum (Pav.) P. Schiller (Ba 12), was more sensitive to UV-B radiation than the non-motile diatoms Dityhim brightwellii (P. West) Grun (Ba 15) and Phaeodactylum tricornutum Bohlin (Ba 16). One week of UV-B radiation 2 h daily (159 J m⁻² day⁻¹), had a dramatic effect on the growth of the dinoflagellates, while the diatoms were nearly unaffected. On the other hand, when given higher intensity of UV-B radiation (312, 468 and 624 J m⁻² day⁻¹) during the initial phase of growth, also the growth of the diatom, D. brightwellii, was inhibited. Not only the growth but also the swimming speed of the dinoflagellates C. aureolum and P. minimum were affected by UV-B radiation. The speed decreased rapidly after 1–2 h of UV-B radiation (312 J m⁻² day⁻¹), and after longer irradiation times the dinoflagellates lost their motility. G. aureolum exposed to UV-B radiation, regained normal speed after two weeks of visible light.     

The significance of the study about the biological effects of solar ultraviolet radiation using the Exposed Facility on the International Space Station.

It is believed that ultraviolet (UV) radiation from the sun participated in events related to the chemical evolution and birth of life on the primitive Earth. Although UV radiation would be also a driving force for the biological evolution of life on Earth, life space of the primitive living organisms would be limited in the UV-shielded place such as in the water at an early stage of the evolution of life. After the formation of stratospheric ozone layer through the production of oxygen by photoautotroph, living organisms were able to expand their domain from water to land. As a result, now, many kinds of living organisms containing human beings are flourishing on the ground. In the near future, increased transmission of harmful solar UV radiation may reach the Earth's surface due to stratospheric ozone layer depletion. In order to learn more about the biological effects of solar UV radiation with or without interruption by the ozone layer, the utilization of an Exposed Facility on the International Space Station is required. Experiments proposed for this facility would provide a tool for the scientific investigation of processes involved in the birth and evolution of life on Earth, and could also demonstrate the importance of protecting the Earth's future environment from future ozone layer depletion.