Metals and low dose IR: Molecular effects of combined exposures using HepG2 cells as a biological model.

Uranium mining sites produce residues rich in metals and radionuclides, that may contaminate all environmental matrices, exposing human and non-human biota to low doses of ionizing radiation (LDIR) and to the chemical toxicity of several metals. To date, experimental and radio-epidemiological studies do not provide conclusive evidence of LDIR induced cancer. However, co-exposures (LDIR plus other contaminants), may increase the risks. To determine the potential for genotoxic effects in human cells induced by the exposure to LDIR plus metals, HEPG2 cells were exposed to different concentrations of a uranium mine effluent for 96 h. DNA damage was evaluated using the comet assay and changes in the expression of tumor suppressor and oncogenes were determined using qPCR. Results show that effluent concentrations higher than 5%, induce significant DNA damage. Also, a significant under-expression of ATM and TP53 genes and a significant overexpression of GADD45a gene was observed. Results show that the exposure to complex mixtures cannot be disregarded, as effects were detected at very low doses. This study highlights the need for further studies to clarify the risks of exposure to LDIR along with other stressors, to fully review the IR exposure risk limits established for human and non-human biota.

Life history traits and genotoxic effects on Daphnia magna exposed to waterborne uranium and to a uranium mine effluent - A transgenerational study.

Assessing the impact of uranium mining industry on aquatic ecosystems near mining areas is critical to ensure the long-term health and sustainability of ecosystem services. As so, a transgenerational study with Daphnia magna has been conducted to perceive to what extent intermittent discharges of uranium mine effluents into watercourses may impact the DNA integrity and life history traits of cladocerans. Organisms were exposed for 48 h to a 2% dilution of an uranium mine effluent (UME) and to a corresponding dose of waterborne uranium (WU) that, according to our preliminary studies, induces significant DNA damage in daphnids. After exposure, organisms were transferred to clean medium, where three successive generations were monitored for genotoxicity and other effects at the individual and population level. Despite some differences between WU and UME data, our results revealed that the negative impacts of the short-term exposure gradually disappeared after placing the organisms in clean medium. These results suggest that, under intermittent stress, daphnids are able to recover, since after the 3rd brood release, DNA damage (measured as DNA strand breaks) is no longer observed and has no significant impact on the detectable life traits of offspring. Although our results indicate that populations of D. magna are not affected by intermittent and highly diluted discharges from uranium mining, aquatic systems under this kind of pressure should not be seen as hazardous-free. Future studies in this field are recommended and these should consider radionuclides in the water column, their accumulation in the sediments and also multiple life stages.

RIBE at an inter-organismic level: A study on genotoxic effects in Daphnia magna exposed to waterborne uranium and a uranium mine effluent.

The induction of RIBE (Radiation Induced Bystander Effect) is a non-target effect of low radiation doses that has already been verified at an inter-organismic level in fish and small mammals. Although the theoretical impact in the field of environmental risk assessment (ERA) is possible, there is a gap of knowledge regarding this phenomenon in invertebrate groups and following environmentally relevant exposures. To understand if RIBE should be considered for ERA of radionuclide-rich wastewaters, we exposed Daphnia magna (<24 h and 5d old) to a 2% diluted uranium mine effluent for 48 h, and to a matching dose of waterborne uranium (55.3 µg L-1). Then the exposed organisms were placed (24 and 48 h) in a clean medium together with non-exposed neonates. The DNA damage observed for the non-exposed organisms was statistically significant after the 24 h cohabitation for both uranium (neonates p = 0.002; 5 d-old daphnids p = <0.001) and uranium mine effluent exposure (only for neonates p = 0.042). After 48 h cohabitation significant results were obtained only for uranium exposure (neonates p = 0.017; 5 d-old daphnids p = 0.013). Although there may be some variability associated to age and exposure duration, the significant DNA damage detected in non-exposed organisms clearly reveals the occurrence of RIBE in D. magna. The data obtained and here presented are a valuable contribution for the discussion about the relevance of RIBE for environmental risk assessment.

Source identification of uranium-containing materials at mine legacy sites in Portugal.

Whilst prior nuclear forensic studies have focused on identifying signatures to distinguish between different uranium deposit types, this paper focuses on providing a scientific basis for source identification of materials from different uranium mine sites within a single region, which can then be potentially used within nuclear forensics. A number of different tools, including gamma spectrometry, alpha spectrometry, mineralogy and major and minor elemental analysis, have been utilised to determine the provenance of uranium mineral samples collected at eight mine sites, located within three different uranium provinces, in Portugal. A radiation survey was initially conducted by foot and/or unmanned aerial vehicle at each site to assist sample collection. The results from each mine site were then compared to determine if individual mine sites could be distinguished based on characteristic elemental and isotopic signatures. Gamma and alpha spectrometry were used to differentiate between samples from different sites and also give an indication of past milling and mining activities. Ore samples from the different mine sites were found to be very similar in terms of gangue and uranium mineralogy. However, rarer minerals or specific impurity elements, such as calcium and copper, did permit some separation of the sites examined. In addition, classification rates using linear discriminant analysis were comparable to those in the literature.

APPLICATION OF DROPLET DETECTORS TO ALPHA RADIATION DETECTION.

Superheated droplet detectors (SDDs) are traditionally employed in the detection of neutrons. In this work the focus is on the detection of alpha particles using C2ClF5 as the target liquid. The alpha-droplet interaction is examined via computational studies, and a geometric model developed to describe the expected detector response. Experiments with alpha-emitting uranium- and samarium-doped SDDs at temperatures of 5-12°C confirm that the event rate is related to the size of the droplets, and are in model agreement for temperatures below 8°C; above this temperature, the acoustic sensitivity is reduced by signal attenuation as a result of the increasing bubble population, for which the addition of an attenuation coefficient restores the agreement with experiment. The results suggest the viability of a SDD-based alpha spectrometer using mono-sized droplets.

Uranium mining wastes: The use of the Fish Embryo Acute Toxicity Test (FET) test to evaluate toxicity and risk of environmental discharge.

Active and abandoned uranium mining sites often create environmentally problematic situations, since they cause the contamination of all environmental matrices (air, soil and water) with stable metals and radionuclides. Due to their cytotoxic, genotoxic and teratogenic properties, the exposure to these contaminants may cause several harmful effects in living organisms. The Fish Embryo Acute Toxicity Test (FET) test was employed to evaluate the genotoxic and teratogenic potential of mine liquid effluents and sludge elutriates from a deactivated uranium mine. The aims were: a) to determine the risk of discharge of such wastes in the environment; b) the effectiveness of the chemical treatment applied to the uranium mine water, which is a standard procedure generally applied to liquid effluents from uranium mines and mills, to reduce its toxicological potential; c) the suitability of the FET test for the evaluation the toxicity of such wastes and the added value of including the evaluation of genotoxicity. Results showed that through the FET test it was possible to determine that both elutriates and effluents are genotoxic and also that the mine effluent is teratogenic at low concentrations. Additionally, liquid effluents and sludge elutriates affect other parameters namely, growth and hatching and that water pH alone played an important role in the hatching process. The inclusion of genotoxicity evaluation in the FET test was crucial to prevent the underestimation of the risks posed by some of the tested effluents/elutriates. Finally, it was possible to conclude that care should be taken when using benchmark values calculated for specific stressors to evaluate the risk posed by uranium mining wastes to freshwater ecosystems, due to their chemical complexity.

Certified reference materials for radionuclides in Bikini Atoll sediment (IAEA-410) and Pacific Ocean sediment (IAEA-412).

The preparation and characterization of certified reference materials (CRMs) for radionuclide content in sediments collected offshore of Bikini Atoll (IAEA-410) and in the open northwest Pacific Ocean (IAEA-412) are described and the results of the certification process are presented. The certified radionuclides include: (40)K, (210)Pb ((210)Po), (226)Ra, (228)Ra, (228)Th, (232)Th, (234)U, (238)U, (239)Pu, (239+240)Pu and (241)Am for IAEA-410 and (40)K, (137)Cs, (210)Pb ((210)Po), (226)Ra, (228)Ra, (228)Th, (232)Th, (235)U, (238)U, (239)Pu, (240)Pu and (239+240)Pu for IAEA-412. The CRMs can be used for quality assurance and quality control purposes in the analysis of radionuclides in sediments, for development and validation of analytical methods and for staff training.

Uranium mining in Portugal: a review of the environmental legacies of the largest mines and environmental and human health impacts.

The history of uranium mining in Portugal during almost one century has followed international demand peaks of both radium and uranium, which in turn were driven by medical, military, and civil applications. Nowadays, following price drop in the 1980s, mining activities decreased and ceased in 2001. The current challenge is to deal with environmental legacies left by old uranium mines, mainly located in Viseu and Guarda districts. In 2001, based on several radiological surveys carried out, the Portuguese government assumed the remediation costs of abandoned mine areas for environmental safety and public health protection. Detailed environmental and public health risk assessments were performed under the scope of studies both requested by the government and by funded research projects. It was found that the existing risks, due to radiological and chemical exposures to metals and radionuclide's, were particularly high at the old milling facilities and mines where in situ and heap leaching of low-grade ore occurred. The different studies, involving both humans and non-human species from different trophic levels, demonstrated the existence of effects at different levels of biological organization (molecular, cellular, tissues, individuals, and populations) and on ecosystem services. To mitigate the risks, the environmental rehabilitation works at the Urgeiriça mine complex are almost complete, while at Cunha Baixa mine, they are presently in progress. These works and environmental improvements achieved and expected are described herein.

Certified Reference Material IAEA-446 for radionuclides in Baltic Sea seaweed.

A Certified Reference Material (CRM) for radionuclides in seaweed (Fucus vesiculosus) from the Baltic Sea (IAEA-446) is described and the results of the certification process are presented. The (40)K, (137)Cs, (234)U and (239+240)Pu radionuclides were certified for this material, and information values for 12 other radionuclides ((90)Sr, (99)Tc, (210)Pb ((210)Po), (226)Ra, (228)Ra, (228)Th, (230)Th, (232)Th, (235)U, (238)U, (239)Pu and (240)Pu) are presented. The CRM can be used for Quality Assurance/Quality Control of analysis of radionuclides in seaweed and other biota samples, as well as for development and validation of analytical methods, and for training purposes.

Final analysis and results of the Phase II SIMPLE dark matter search.

We report the final results of the Phase II SIMPLE measurements, comprising two run stages of 15 superheated droplet detectors each, with the second stage including an improved neutron shielding. The analyses include a refined signal analysis, and revised nucleation efficiency based on a reanalysis of previously reported monochromatic neutron irradiations. The combined results yield a contour minimum of σp=5.7×10(-3) pb at 35 GeV/c2 in the spin-dependent sector of weakly interacting massive particle (WIMP) proton interactions, the most restrictive to date for MW}≤60 GeV/c2 from a direct search experiment and overlapping, for the first time, with results previously obtained only indirectly. In the spin-independent sector, a minimum of 4.7×10(-6) pb at 35 GeV/c2 is achieved, with the exclusion contour challenging a significant part of the light mass WIMP region of current interest.

Characteristics of Lactobacillus and Gardnerella vaginalis from women with or without bacterial vaginosis and their relationships in gnotobiotic mice.

The objectives of the present study were to evaluate in vitro the production of antagonistic compounds against Gardnerella vaginalis by Lactobacillus strains isolated from women with or without bacterial vaginosis (BV), and to select one of the better Lactobacillus producers of such a substance to be tested in vivo using a gnotobiotic animal model challenged with one of the more sensitive G. vaginalis isolates. A total of 24 isolates from women with and without BV were identified as G. vaginalis. A higher frequency (P<0.05) of this bacterium was observed in women with BV (56.7%) when compared to healthy women (17.6%). A total of 86 strains of Lactobacillus were obtained from healthy women and women with BV. Lactobacillus strains were more frequently present (P<0.05) in healthy women (97.5%) than in women with BV (76.7%). Lactobacillus crispatus was the predominating strain in both healthy women and women with BV. Lactobacillus jensenii, Lactobacillus johnsonii, Lactobacillus gasseri and Lactobacillus vaginalis were isolated with an intermediate frequency in the two groups. In vitro antagonism assays were performed using as indicators 17 reference strains and the G. vaginalis strains isolated from women with BV and from healthy women. Lactobacillus isolated from healthy women showed the higher antagonistic activity against all the indicator strains when compared with isolates from women with BV. Concerning the indicator strains, G. vaginalis found in women with BV was more resistant to the antagonism, particularly when Lactobacillus isolates from women with BV were used as producer strains. A high vaginal population level of G. vaginalis was obtained by intravaginal inoculation of germ-free mice, and this colonization was accompanied by vaginal histopathological lesions. A tenfold decrease in vaginal population level of G. vaginalis and a reduction of histological lesions were observed when the pathogenic challenge was performed in mice previously monoassociated with an L. johnsonii strain. Concluding, results of the present study suggest that progression of G. vaginalis-associated BV depends in part on a simultaneous presence of Lactobacillus populations with a low antagonistic capacity and of a G. vaginalis strain with a high resistance to this antagonism. The results could also explain why G. vaginalis is frequently found in the vaginal ecosystem of healthy women.

Radioactivity from Fukushima nuclear accident detected in Lisbon, Portugal.

The radioactivity released from the Fukushima Dai-ichi nuclear accident was transported around the globe by atmospheric processes. Several artificial radionuclides were detected and measured in aerosols and atmospheric surface depositions in the Lisbon area during late March and early April 2011. The highest concentrations measured in aerosols were those of particulate (131)I, 1.39 ± 0.08 mBq m(-3). Cesium-134, (137)Cs and (132)Te were also determined but at lower concentrations. The total atmospheric depositions on the ground were higher on the first week of April with values for (131)I, (134)Cs, and (137)Cs of 0.92 ± 0.11, 0.59 ± 0.06, and 0.62 ± 0.12 Bq m(-2), respectively. The four artificial radionuclides measurable, (131)I, (132)Te (134)Cs, and (137)Cs, caused little radiation exposure to the members of the public, that was five orders of magnitude lower than the ionizing radiation effective dose limits for members of the public for one year (1 mSv y(-1)).

Tracking of airborne radionuclides from the damaged Fukushima Dai-ichi nuclear reactors by European networks.

Radioactive emissions into the atmosphere from the damaged reactors of the Fukushima Dai-ichi nuclear power plant (NPP) started on March 12th, 2011. Among the various radionuclides released, iodine-131 ((131)I) and cesium isotopes ((137)Cs and (134)Cs) were transported across the Pacific toward the North American continent and reached Europe despite dispersion and washout along the route of the contaminated air masses. In Europe, the first signs of the releases were detected 7 days later while the first peak of activity level was observed between March 28th and March 30th. Time variations over a 20-day period and spatial variations across more than 150 sampling locations in Europe made it possible to characterize the contaminated air masses. After the Chernobyl accident, only a few measurements of the gaseous (131)I fraction were conducted compared to the number of measurements for the particulate fraction. Several studies had already pointed out the importance of the gaseous (131)I and the large underestimation of the total (131)I airborne activity level, and subsequent calculations of inhalation dose, if neglected. The measurements made across Europe following the releases from the Fukushima NPP reactors have provided a significant amount of new data on the ratio of the gaseous (131)I fraction to total (131)I, both on a spatial scale and its temporal variation. It can be pointed out that during the Fukushima event, the (134)Cs to (137)Cs ratio proved to be different from that observed after the Chernobyl accident. The data set provided in this paper is the most comprehensive survey of the main relevant airborne radionuclides from the Fukushima reactors, measured across Europe. A rough estimate of the total (131)I inventory that has passed over Europe during this period was <1% of the released amount. According to the measurements, airborne activity levels remain of no concern for public health in Europe.

A new Certified Reference Material for radionuclides in Irish sea sediment (IAEA-385).

A new Certified Reference Material (CRM) for radionuclides in sediment (IAEA-385) is described and the results of the certification process are presented. Eleven radionuclides ((40)K, (137)Cs, (226)Ra, (228)Ra, (230)Th, (232)Th, (234)U, (238)U, (238)Pu, (239+240)Pu and (241)Am) have been certified and information mass activities with 95% confidence intervals are given for seven other radionuclides ((90)Sr, (210)Pb((210)Po), (235)U, (239)Pu, (240)Pu and (241)Pu). Results for less frequently reported radionuclides ((60)Co, (99)Tc, (134)Cs, (155)Eu, (224)Ra and (239)Np) and information on some activity and mass ratios are also reported. The CRM can be used for quality assurance/quality control of the analysis of radionuclides in sediment samples, for the development and validation of analytical methods and for training purposes.

Radioactivity in the environment around past radium and uranium mining sites of Portugal.

Measurements of ambient radiation doses and determination of radionuclide concentrations in mining waste and soils were performed in 60 areas of former radium and uranium mining. In several places, mining waste and low-grade uranium ore left on the surface contain radioactivity above regional background. Most of the former mining sites present no enhanced radionuclide concentrations. However, in the mining facilities where the radioactive ore was chemically extracted, mill tailings contain materials with elevated levels of radioactivity, up to 200 times the levels in unaffected soils of the region. Mud from neutralization ponds used to treat acid mine waters contains also elevated radionuclide concentrations. Furthermore, depending on the type of waste, the radioelement composition varies. Environmental rehabilitation measures shall take these differences into account in order to prevent in the long term the radioactive contamination of agriculture soils and water resources, and to ensure adequate radiological protection to the public and to the environment.

Certified reference material for radionuclides in fish flesh sample IAEA-414 (mixed fish from the Irish Sea and North Sea).

A certified reference material (CRM) for radionuclides in fish sample IAEA-414 (mixed fish from the Irish Sea and North Seas) is described and the results of the certification process are presented. Nine radionuclides (40K, 137Cs, 232Th, 234U, 235U, 238U, 238Pu, 239+240Pu and 241Am) were certified for this material. Information on massic activities with 95% confidence intervals is given for six other radionuclides (90Sr, 210Pb(210Po), 226Ra, 239Pu, 240Pu 241Pu). Less frequently reported radionuclides (99Tc, 129I, 228Th, 230Th and 237Np) and information on some activity and mass ratios are also included. The CRM can be used for quality assurance/quality control of the analysis of radionuclides in fish sample, for the development and validation of analytical methods and for training purposes. The material is available from IAEA, Vienna, in 100 g units.

Toxaphene residues from cotton fields in soils and in the coastal environment of Nicaragua.

Toxaphene (camphechlor) was intensively used in the cotton growing fields of Nicaragua for decades with application rates as high as 31 kg ha(-1) in 1985. Although the use of this compound has recently been discontinued in the country, its intensive use in the past and its long persistence in soil allowed for the build up of large reservoirs of toxaphene in agriculture soils and a wide dispersal of residues in the environment. Measurements of toxaphene in coastal areas on the coast of the Pacific Ocean show that environmental concentrations are particularly high in the district of Chinandega, the traditional cotton growing region. Toxaphene residues measured in soils attained 44 microg g(-1) (dry weight) while concentrations in lagoon sediments attained 6.9 microg g(-1) (dry weight) near the mouth of the rivers flowing across the agricultural region. Measurements in aquatic biota showed concentrations as high as 1.6 microg g(-1) (dry weight) in the soft tissues of clams. The toxaphene reservoir in soils combined with the obvious persistence of this compound in soils and lagoon sediments allows predicting that toxaphene will remain in the coastal ecosystem at relatively high concentrations for many years. Toxic effects in lagoon fauna are likely to be observed especially in benthic species that may recycle this compound from sediments. Consumption of seafood, in particular of clams (Anadara spp.) from the more contaminated areas, may expose the population to unacceptably high intake of toxaphene, 30 microg d(-1) per person, with the diet.

Fate of 14C-chlorpyrifos in the tropical estuarine environment.

The distribution and fate of 14C-chlorpyrifos were investigated in microcosms simulating the conditions of the tropical estuarine environment of North Vietnam. The microcosms containing brackish water, sediment, clams (Meretrix meretrix) and green algae (Gracilaria verucosa) from the Red River estuary, were maintained for 30 days. The results show that chlorpyrifos released into the water was rapidly adsorbed onto sediment. However, only 1-2% of the initial amount of 14C-chlorpyrifos could be detected in the sediment by the end of the experiment. The accumulation of chlorpyrifos in fauna and flora attained, respectively, a maximum of 5.8% and 2.2% of the initial activity observed at days 3 and 2 after application. The compound 3,4,5-trichloro-2pyridinol (TCP) was the major transformation product of chlorpyrifos found in the microcosm. TCP accumulated in the clam's soft tissues, and, 3 days after application, reached a maximum of 0.5% of the total 14C-activity. The balance of the 14C-activity at the end of the experiment suggests that the main loss of the insecticide from the system was through volatilisation of chlorpyrifos and escape to the atmosphere. The persistence half-time of the compound in the aquatic microcosms was computed at 5 days.

Distribution, fate and effects of pesticide residues in tropical coastal lagoons of northwestern Mexico.

Analyses of pesticide residues in sediments, water and biota of the Altata-Ensenada del Pabellon coastal lagoon system in Sinaloa, Mexico, showed the presence of organochlorine and organophosphorus compounds. For all the compounds analysed, concentrations in sediments were higher near the points of water discharge from ponds and drainage from the agricultural area. Among the organochlorines, total DDTs displayed the largest sedimentary reservoir, followed by total endosulfans and chlorpyrifos. In sediments, as well as in biota, pp'-DDT concentrations were lower than those of DDT metabolites, which confirms the reduction in the previous massive use of this compound in agriculture. Endosulfan is currently used in the region and endosulfan residues in lagoon sediments attained levels considered to be toxic to meiofauna, therefore constituting an ecological risk to lagoon ecosystems. There was a large sedimentary reservoir of chlorpyrifos but its ecotoxicological risk is difficult to assess due to lack of adequate comparative data. Nevertheless, concentrations of chlorpyrifos which approach acute toxic levels for shrimp were recorded in lagoon water. This suggests that drainage from agricultural fields during high runoff may, on occasion, cause mass mortality of shrimp and fish Organophosphorus pesticides are widely regarded to degrade very rapidly in aquatic systems. Experimental research performed with 14C-labelled chlorpyrifos and parathion has shown that they may be stabilised for relatively long periods of time through sediment-water partitioning. This extension of their environmental half-lives increases their potential for impacting on coastal ecosystems. Since organophosphorus pesticides are highly toxic for aquatic organisms at concentrations generally lower than organochlorines, their presence in the lagoon water and sediment is a matter for much concern. The increased use of tropical coastal lagoons for shrimp and fish farming requires the implementation of environmental management practices to protect these ecosystems from the impact of agricultural activities. Environmental management aims to preserve the ecosystem integrity of these coastal lagoons and, through improving the water quality, to allow the development of aquaculture and reduce human exposure to pesticide residues in food.

Evaluation of recombinant Leptospira antigen-based enzyme-linked immunosorbent assays for the serodiagnosis of leptospirosis.

There is an urgent need for development of new serodiagnostic strategies for leptospirosis, an emerging zoonosis with worldwide distribution. We have evaluated the diagnostic utility of five recombinant antigens in enzyme-linked immunosorbent assays (ELISAs) for serodiagnosis of leptospirosis. Sera from 50 healthy residents of a high-incidence region were used to determine cutoff values for 96% specificity. In paired sera from 50 cases of leptospirosis confirmed by the microscopic agglutination test, immunoglobulin G (IgG) but not IgM reacted with the recombinant leptospiral proteins. The recombinant LipL32 IgG ELISA had the highest sensitivities in the acute (56%) and convalescent (94%) phases of leptospirosis. ELISAs based on recombinant OmpL1, LipL41, and Hsp58 had sensitivities of 16, 24, and 18% during the acute phase and 72, 44, and 32% during convalescence, respectively. Compared to sera from healthy individuals, patient sera did not react significantly with recombinant LipL36 (P > 0.05). Recombinant LipL32 IgG ELISA demonstrated 95% specificity among 100 healthy individuals, and specificities ranging from 90 to 97% among 30 dengue patients, 30 hepatitis patients, and 16 patients with diseases initially thought to be leptospirosis. Among 39 Venereal Disease Research Laboratory test-positive individuals and 30 Lyme disease patients, 13 and 23% of sera, respectively, reacted positively with the rLipL32 antigen. These findings indicate that rLipL32 may be an useful antigen for the serodiagnosis of leptospirosis.