Effects of acute γ-irradiation on community structure of the aquatic microbial microcosm

To characterise indirect effects of ionising radiation on aquatic microbial communities, effects of acute γ-irradiation were investigated in a microcosm consisting of populations of green algae (Chlorella sp. and Scenedesmus sp.) and a blue-green alga (Tolypothrix sp.) as producer; a ciliate protozoan (Cyclidium glaucoma), rotifers (Lecane sp. and Philodina sp.) and an oligochaete (Aeolosoma hemprichi) as consumer; and more than four species of bacteria as decomposers. Population changes in the constituent organisms were observed over 160 days after irradiation. Prokaryotic community structure was also examined by denaturing gradient gel electrophoresis (DGGE) of 16S rDNA. Principle response curve analysis revealed that the populations of the microcosm as a whole were not significantly affected at 100 Gy while they were adversely affected at 500-5000 Gy in a dose-dependent manner. However, some effects on each population, including each bacterial population detected by DGGE, did not depend on radiation doses, and some populations in the irradiated microcosm were larger than those of the control. These unexpected results are regarded as indirect effects through interspecies interactions, and possible mechanisms are proposed originating from population changes in other organisms co-existing in the microcosm. For example, some indirect effects on consumers and decomposers likely arose from interspecies competition within each trophic level. It is also likely that prey-predator relationships between producers and consumers caused some indirect effects on producers.     

Assessment of natural or enhanced in situ bioremediation at a chlorinated solvent-contaminated aquifer in Italy: a microcosm study

A microcosm study was used to assess the potential for in situ natural or enhanced bioremediation at a chloroethane- (i.e., tetrachloroethane, TeCA) and chloroethene-contaminated (i.e., tetrachloroethene, PCE; trichloroethene, TCE) groundwater in Northern Italy. All the live microcosms were positive for dechlorination, indicating the presence of an active native dechlorinating population in the subsurface. All the tested electron donors (i.e., yeast extract, lactate, butyrate, hydrogen) promoted enhanced dechlorination of chlorinated contaminants. Lactate- and butyrate-amended microcosms performed the best, and also dechlorinated the solvents past cis-dichloroethene (cis-DCE). The microcosm bioaugmented with a PCE-dechlorinating mixed culture containing Dehalococcoides spp. dechlorinated groundwater contaminants to DCE, vinyl chloride (VC), and ethene (ETH). In conclusion, results from this microcosm study indicate the potential for enhancing full dechlorination at the contaminated site, through a proper addition of a suitable electron donor (e.g., lactate or butyrate) and/or through bioaugmentation with a Dehalococcoides-containing culture.     

Effects of acute γ-irradiation on the aquatic microbial microcosm in comparison with chemicals

Effects of acute γ-irradiation were investigated in the aquatic microcosm consisting of green algae (Chlorella sp. and Scenedesmus sp.) and a blue–green alga (Tolypothrix sp.) as producers; an oligochaete (Aeolosoma hemprichi), rotifers (Lecane sp. and Philodina sp.) and a ciliate protozoan (Cyclidium glaucoma) as consumers; and more than four species of bacteria as decomposers. At 100 Gy, populations were not affected in any taxa. At 500–5000 Gy, one or three taxa died out and populations of two or three taxa decreased over time, while that of Tolypothrix sp. increased. This Tolypothrix sp. increase was likely an indirect effect due to interspecies interactions. The principal response curve analysis revealed that the main trend of the effects was a dose-dependent population decrease. For a better understanding of radiation risks in aquatic microbial communities, effect doses of γ-rays compared with copper, herbicides and detergents were evaluated using the radiochemoecological conceptual model and the effect index for microcosm.     

Cytogenetic dose-response and adaptive response in cells of ungulate species exposed to ionizing radiation

In the studies reported here, the micronucleus assay, a common cytogenetic technique, was used to examine the dose-responses in fibroblasts from three ungulate species (white-tailed deer, woodland caribou, and Indian muntjac) exposed to high doses of ionizing radiation (1-4 Gy of (60)Co gamma radiation). This assay was also used to examine the effects of exposure to low doses (1-100 mGy) typical of what these species experience in a year from natural and anthropogenic environmental sources. An adaptive response, defined as the induction of resistance to a stressor by a prior exposure to a small "adapting" stress, was observed after exposure to low doses. This work indicates that very small doses are protective for the endpoint examined. The same level of protection was seen at all adapting doses, including 1 radiation track per cell, the lowest possible cellular dose. These results are consistent with other studies in a wide variety of organisms that demonstrate a protective effect of low doses at both cellular and whole-organism levels. This implies that environmental regulations predicated on the idea that even the smallest dose of radiation carries a quantifiable risk of direct adverse consequences to the exposed organism require further examination. Cytogenetic assays provide affordable and feasible biological effects-based alternatives that are more biologically relevant than traditional contaminant concentration-based radioecological risk assessment.     

Challenges in radioecology

Today, radioecology covers a broad scientific field; from the source to long term environmental impact from ionizing radiation. To summarize key challenges within radioecology, the present paper focuses upon knowledge gaps related to processes, mechanisms and variables contributing most to the overall uncertainties in environmental impact assessments. A series of sources related to the nuclear weapon cycle and the civil nuclear cycle has contributed, is still contributing or can potentially contribute to release of radionuclides to the environment in the future. The speciation of most radionuclides depends on the source and release conditions, and will influence ecosystem transport, biological uptake, doses and effects in flora and fauna. Radionuclides may also co-occur in contaminant mixtures (e.g., metals, organics), which potentially could lead to synergisms or antagonisms. Thus, challenges associated with the links between the source or release term – radionuclide speciation – ecosystem transfer – exposure – response relationships are highlighted.     

Low dose beta-emitter source induces sexual reproduction instead of fragmentation in an earthworm, Enchytraeus japonensis

We examined whether background radiation, or radiation at a slightly higher level, plays a role in the reproduction of a terrestrial earthworm. Enchytraeus japonensis a recently described terrestrial oligochaete, reproduces asexually by fragmentation and subsequent regeneration. Following radiation exposure in which the worms were subjected to a 32P beta-emitter source at 15 times the background dose rate (4.5 microGy/h), a statistically significant decrease in the number of fragmentations was observed as compared with the sham controls. At that time, in a stained preparation with haematoxylin and eosin (HE), sexual reproduction occurred instead of asexual fragmentation, and mature oocytes were observed in the body of grown worms. However, increasing the radiation dose rate by 30 microGy/h resulted in the complete disappearance of the radiation-induced effects, i.e., fragmentation again occurred after 14 h. The results of this study indicate that a lower dose of radiation may be essential to achieve sexual reproduction, inducing an inhibition of fragmentation (asexual reproduction), but at higher, more cytotoxic doses of radiation these effects are negated.     

Ecological impacts of umbrella effects of radiation on the individual members

In order to study the interactions in a model aquatic microcosm, an individual-based computer simulation model was developed. The microcosm consists of Euglena gracilis as an autotroph algae, Tetrahymena thermophila as a heterotroph protozoa and Escherichia coli as a saprotroph bacteria. There exists a strong interaction between Tetrahymena and E. coli as the first is the predator of the second. Ecological toxicity tests were conducted to test the population level impacts of the biological effects of radiation and toxicants on the lethality and mobility factors that influence directly or indirectly growth and reproduction. Radiological effects on lethality of E. coli individuals were translated to the reduction of the equilibrium population of Tetrahymena. A synergistic effect at the community level was also observed by the simulation of a combined exposure of radiation and a toxicant which reduced the feeding efficiency of Tetrahymena.     

Genomic instability in human osteoblast cells after exposure to depleted uranium: delayed lethality and micronuclei formation

It is known that radiation can induce a transmissible persistent destabilization of the genome. We have established an in vitro cellular model using HOS cells to investigate whether genomic instability plays a role in depleted uranium (DU)-induced effects. Transmissible genomic instability, manifested in the progeny of cells exposed to ionizing radiation, has been characterized by de novo chromosomal aberrations, gene mutations, and an enhanced death rate. Cell lethality and micronuclei formation were measured at various times after exposure to DU, Ni, or gamma radiation. Following a prompt, concentration-dependent acute response for both endpoints, there was de novo genomic instability in progeny cells. Delayed reproductive death was observed for many generations (36 days, 30 population doublings) following exposure to DU, Ni, or gamma radiation. While DU stimulated delayed production of micronuclei up to 36 days after exposure, levels in cells exposed to gamma-radiation or Ni returned to normal after 12 days. There was also a persistent increase in micronuclei in all clones isolated from cells that had been exposed to nontoxic concentrations of DU. While clones isolated from gamma-irradiated cells (at doses equitoxic to metal exposure) generally demonstrated an increase in micronuclei, most clonal progeny of Ni-exposed cells did not. These studies demonstrate that DU exposure in vitro results in genomic instability manifested as delayed reproductive death and micronuclei formation.     

Analysis of rodent populations in technogenically transformed areas (with reference to <Emphasis Type="Italic">Apodemus</Emphasis> (<Emphasis Type="Italic">S.</Emphasis>) <Emphasis Type="Italic">uralensis</Emphasis> from the EURT zone)

It is shown that the response of rodent populations to acute and chronic irradiation depends on its functional structure, i.e., on specific features of animals with two alternative types of ontogenetic development. Upon acute irradiation, sexually immature young of the year (animals with the second type of ontogeny) are most radioresistant. Exposure to chronic irradiation, as in the zone of the Eastern Ural Radioactive Trace (EURT), leads to an increase in the proportion of mature young of the year (animals with the first type of ontogeny), which are the most radiosensitive part of the population. The abundance and fecundity of mice in the impact zone are consistently higher than in the background zone, which improves the adaptive potential of the population. The role of species ecological specialization and configuration of the contaminated zone in the formation of migrant rodent population is emphasized. It is concluded that a high migration activity allows the pigmy wood mouse (a radiosensitive species) to avoid long-term radiation exposure.     

Comparative radiation impact on biota and man in the area affected by the accident at the Chernobyl nuclear power plant

A methodological approach for a comparative assessment of ionising radiation effects on man and non-human species, based on the use of Radiation Impact Factor (RIF) - ratios of actual exposure doses to biota species and man to critical dose is described. As such doses, radiation safety standards limiting radiation exposure of man and doses at which radiobiological effects in non-human species were not observed after the Chernobyl accident, were employed. For the study area within the 30km ChNPP zone dose burdens to 10 reference biota groups and the population (with and without evacuation) and the corresponding RIFs were calculated. It has been found that in 1986 (early period after the accident) the emergency radiation standards for man do not guarantee adequate protection of the environment, some species of which could be affected more than man. In 1991 RIFs for man were considerably (by factor of 20.0-1.1 x 10(5)) higher compared with those for selected non-human species. Thus, for the long term after the accident radiation safety standards for man are shown to ensure radiation safety for biota as well.     

Problems of sustainable development and ecological indication in radioactively contaminated areas

Lands that have become unusable because of contamination with long-lived artificial radionuclides require special measures aimed at their monitoring and rehabilitation. In such areas, specific processes in the biota are associated with the direct effect of ionizing radiation on some species, secondary changes depending on the ecology of organisms trophically connected with these species, and side effects explained by the regime of isolation established in radiation reserves. The dynamics of floras and faunas in radiation reserves are determined by successional changes in plant communities, migration of radionuclides, and the protective status of these territories.     

Toxicological effects of disinfections using sodium hypochlorite on aquatic organisms and its contribution to AOX formation in hospital wastewater

Sodium hypochlorite (NaOCl) is often used for disinfecting hospital wastewater in order to prevent the spread of pathogenic microorganisms, causal agents of nosocomial infectious diseases. Chlorine disinfectants in wastewater react with organic matters, giving rise to organic chlorine compounds such as AOX (halogenated organic compounds adsorbable on activated carbon), which are toxic for aquatic organisms and are persistent environmental contaminants. The aim of this study was to evaluate the toxicity on aquatic organisms of hospital wastewater from services using NaOCl in pre-chlorination. Wastewater samples from the infectious and tropical diseases department of a hospital of a large city in southeast of France were collected. Three samples per day were collected in the connecting well department at 9 a.m., 1 p.m. and 5 p.m. during 8 days from 13 March to 22 March 2001, and a mixture was made at 6 p.m. with the three samples in order to obtain a representative sample for the day. The toxicity test comprised the 24-h EC50 on Daphnia magna and a bioluminescence assay using Vibrio fischeri photobacteria. Fecal coliforms and physicochemical analyses such as total organic carbon (TOC), chloride, AOX, total suspended solids (TSS) and chemical oxygen demand (COD) were carried out. Wastewater samples highlighted considerable acute toxicity on D. magna and V. fischeri photobacteria. However, low most probable numbers (MPN), ranging from <3 to 2400 for 100 ml, were detected for fecal coliforms. Statistical analysis, with a confidence interval of 95%, gave a strong linear regression assessed with r=0.98 between AOX concentrations and EC50 (TU) on daphnia. The identification of an ideal concentration of NaOCl in disinfecting hospital wastewater, i.e. its non-observed effect concentration (NOEC) on algae and D. magna, seems to be a research issue that could facilitate the control of AOX toxicity effects on aquatic organisms. Therefore, it would be necessary to monitor the biocide properties of NaOCl on fecal coliforms at various doses and its toxicity effects on aquatic organisms.     

Choosing an alpha radiation weighting factor for doses to non-human biota

The risk to non-human biota from exposure to ionizing radiation is of current international interest. In calculating radiation doses to humans, it is common to multiply the absorbed dose by a factor to account for the relative biological effectiveness (RBE) of the radiation type. However, there is no international consensus on the appropriate value of such a factor for weighting doses to non-human biota. This paper summarizes our review of the literature on experimentally determined RBEs for internally deposited alpha-emitting radionuclides. The relevancy of each experimental result in selecting a radiation weighting factor for doses from alpha particles in biota was judged on the basis of criteria established a priori. We recommend a nominal alpha radiation weighting factor of 5 for population-relevant deterministic and stochastic endpoints, but to reflect the limitations in the experimental data, uncertainty ranges of 1-10 and 1-20 were selected for population-relevant deterministic and stochastic endpoints, respectively.     

Fractionation of radionuclide species in the environment

Naturally occurring and artificially produced radionuclides in the environment may be present in different physico-chemical forms (i.e., radionuclide species) varying in size (nominal molecular mass), charge properties and valence, oxidation state, structure and morphology, density, degree of complexation, etc. Low molecular mass (LMM) species are believed to be mobile and potentially bioavailable, while high molecular mass (HMM) species such as colloids, polymers, pseudocolloids and particles are considered inert. Due to time-dependent transformation processes such as mobilisation of radionuclide species from solid phases or interactions of mobile and reactive radionuclide species with components in soils and sediments, the original distribution of radionuclides deposited in ecosystems will change over time. To assess the environmental impact from radionuclide contamination, information on radionuclide species deposited, interactions within affected ecosystems and the time-dependent distribution of radionuclide species influencing mobility and biological uptake is essential. The development of speciation techniques to characterize radionuclide species in waters, soils and sediments should therefore be essential for improving the prediction power of impact and risk assessment models. The present paper reviews available fractionation techniques which can be utilised for radionuclide speciation purposes.     

Nematode species at risk--a metric to assess pollution in soft sediments of freshwaters

Soft sediments are often highly polluted as many of the toxic chemicals introduced into surface waters bind to settling particles. The resulting accumulation of pollutants in the sediments poses a risk for benthic communities. However, pollution induced changes in benthic communities have been difficult to determine when using macro-invertebrates as bioindicators, as these organisms are often absent in soft sediment. The present study therefore examined the ability of meiofaunal organisms, specifically, nematodes, to assess the ecological status of soft sediments. Over a 9-year period, nematode communities present in sediments collected from large rivers and lake Constance in Germany were studied. These sediments showed a large range of physico-chemical properties and anthropogenic contamination. After the degree of metal and organic contamination was translated into ecotoxicologically more relevant toxic units (TUs), multivariate methods were used to classify nematode taxa in species at risk (NemaSPEAR) or not at risk (NemaSPE(not)AR). This approach clearly distinguished the influence of sediment texture from that of the toxic potential of the samples and thus allowed classification of the nematode species according to their sensitivity to or tolerance of toxic stress. Two indices, expressing the proportion of species at risk within a sample (NemaSPEAR[%](metal), NemaSPEAR[%](organic)), were calculated from independent data sets obtained in field and experimental studies and showed good correlations with the toxic potential (field data) or chemical concentrations (microcosm data). NemaSPEAR[%] indices for metal and organic pollution were therefore judged to be suitable for assessing the impact of chemical contamination of freshwater soft sediments.     

Adaptive response in frogs chronically exposed to low doses of ionizing radiation in the environment

Using the micronucleus assay, decreased levels of DNA damage were found after high dose ionizing radiation exposure of liver cells taken from frogs inhabiting a natural environment with above-background levels of ionizing radiation, compared to cells taken from frogs inhabiting background areas. The data obtained from a small number of animals suggest that stress present in the above-background environment could induce an adaptive response to ionizing radiation. This study did not reveal harmful effects of exposure to low levels of radioactivity. On the contrary, stress present in the above-background area may serve to enhance cellular defense mechanisms.     

Multiple stressor effects of radiation and metals in salmon (Salmo salar)

These experiments were designed to look at the cellular effects in key organs in Atlantic salmon (Salmo salar) after exposure in vivo to radiation and subtoxic levels of aluminum (Al) and cadmium (Cd), alone or in combination. Salmon (25g) were exposed to a single 0.5Gy dose of gamma-irradiation in water containing Cd, Al or Cd+Al. Three fish per group were sacrificed after 1h and the liver, pronephros, fin and gill of each was dissected. Small explants of each tissue were set up. After 2 days, the culture medium was harvested and filtered then placed on a reporter cell line for determination of stress signal activity (bystander effects). Radiation in combination with Cd and/or Al, caused bystander effects in tissues harvested from in vivo exposed salmon. The effects vary between different organs and are not consistently additive or synergistic for a given treatment. Tissue type appears to be critical. Liver cultures produce a toxic factor which is lethal to reporter cells, and therefore no liver data could be obtained. It is hoped that this stress signal response will prove to be a useful indicator of environmental stress in species inhabiting aquatic ecosystems.     

Radiation-induced adaptive response in fish cell lines

There is considerable interest at present in low-dose radiation effects in non-human species. In this study gamma radiation-induced adaptive response, a low-dose radiation effect, was examined in three fish cell lines, (CHSE-214 (Chinook salmon), RTG-2 (rainbow trout) and ZEB-2J (zebrafish)). Cell survival after exposure to direct radiation with or without a 0.1 Gy priming dose, was determined using the colony forming assay for each cell line. Additionally, the occurrence of a bystander effect was examined by measuring the effect of irradiated cell culture medium from the fish cell lines on unexposed reporter cells. A non-linear dose response was observed for all cell lines. ZEB-2J cells were very sensitive to low doses and a hyper-radiosensitive (HRS) response was observed for doses <0.5 Gy. A typical protective adaptive response was not detected in any of the three fish cell lines tested. Rather, it was found that pre-exposure of these cells to 0.1 Gy radiation sensitized the cells to subsequent high doses. In CHSE-214 cells, increased sensitivity to subsequent high doses of radiation was observed when the priming and challenge doses were separated by 4 h; however, this sensitizing effect was no longer present when the interval between doses was greater than 8 h. Additionally, a "protective" bystander response was observed in these cell lines; exposure to irradiated medium from fish cells caused increased cloning efficiency in unirradiated reporter cells. The data confirm previous conclusions for mammalian cells that the adaptive response and bystander effect are inversely correlated and contrary to expectations probably have different underlying mechanisms.     

Species-Specific Features of Inter- and Intrapopulation Variation in the Level of Chromosome Instability in Rodents

Studies on the common vole Microtus arvalis revealed a distinct cytogenetic response to the influence of ionizing radiation, a wide range of variation in the group average indices of chromosome instability beyond the zone of technogenic impact, and the influence of natural factors (viral infections and fluctuations of population size) on the frequency of chromosome aberrations. In the northern mole vole Ellobius talpinus, neither the mutagenic effect of ionizing radiation nor interpopulation differences in the level of chromosome aberrations in the absence of anthropogenic impact were detected. Variation of genome instability of the rodents is significantly contributed to by species demography and population cycles.     

Adaptation of greater plantain, <Emphasis Type="Italic">Plantago major</Emphasis> L., to long-term radiation and chemical exposure

A comparative study of the greater plantain seed progeny was performed with samples from cenopopulations growing for a long time under conditions of radioactive contamination (in the Eastern Ural Radioactive Trace, EURT) or chemical pollution (in the impact zone of the Nizhny Tagil Iron and Steel Works, NTMK). The progeny of plants from the NTMK zone had low viability but proved to be resistant to the additional impact of a “new” factor (acute γ-irradiation) as well as of the “habitual” factor (heavy metal toxicity). Plantain seeds from the EURT area showed high viability and low heavy metal and radiation resistance; i.e., no preadaptation effect was revealed. In experiments on growing plants from different cenopopulations in plot culture, samples from the EURT zone were characterized mainly by morphoses of generative organs, while samples from the NTMK area, by morphoses of vegetative organs.