On the problem of radioadaptation in small mammals (ecological specialization of species, radioresistance, hemopoiesis, immunity)

The data illustrating ambivalent nature of radioresistance of rodents having different ecological specialization: wood mice (Apodemus uralensis Pallas 1811) and mole-voles (Ellobius talpinus Pallas 1770) to acute (laboratory experiment) and chronic (long-term inhabiting the East Ural Radioactive Trace - EURT) radiation action are presented. These species are greatly distinguished by a way of life, a migratory activity, an average life span etc. Numerous immunological, hematological disturbances were found in more radioresistant wood mice inhabiting in radioactively-polluted biogeocenosis (density of pollution 90Sr - 18.5 MBq/m2 = 500 Ci/km2). On the contrary there are no pathological shifts in more radiosensitive mole-voles from the EURT zone (density of pollution 90Sr - 37 MBq/m2 = 1000 Ci/km2). Comparative analysis of the hematological and immunological parameters and absorbed dose loads in rodents from radiocontaminated environment testify about the possibility of radioadaptation in a series of generations in Ellobius talpinus Pall. The leading role in development of radioadaptation of mole-voles belongs to ecophysiological peculiarities of the species.     

Changes in liver of Apodemus sylvaticus mice from East-Ural radioactive territories

Male Microtus arvalis mice from radionuclide polluted territories in the South Urals (90Sr activity of ground--0.2, 2, 500, 800 Ci/km2) were used. The 90Sr content in a mouse bony skeleton was determined. Carbohydrate, energy and protein metabolism, lipid peroxidation processes in liver were studied. The intensification of protein and carbohydrate enzymes. On the increase of antioxidant activity we found the reduction of lipid peroxidation processes. The decrease of microsome oxidoreductases was found. Possible mechanisms of the observed effects are discussed.     

The cytogenetic consequences of chronic irradiation in rodent populations inhabiting the Eastern Urals Radioactive Trace zone

In bone marrow cells of rodents (Apodemus (Sylvaemus) uralensis Pall., 1811, Apodemus agrarius Pall., 1771) inhabiting the Eastern Urals Radioactive Trace (EURT) zone (Kyshtym radiation accident 1957) and adjacent areas of Urals, the chromosome instability and 90Sr accumulation in bones were investigated. Intensive mutagenic process in both species from impact plots (the soil pollution by 90Sr 2322-16690 kBq/m2) was found. Significant positive correlation of aberrant cells frequencies and 90Sr was shown. Possible causes of the lack of resistance to long-term mutagenic factor (over 100 generations since 50 years from the accident) such as migration of animals and specific configuration of the EURT zone (narrow extended territory with sharply falling gradient of radionuclide pollution), which considerably decrease the probability that certain changes will be fixed and inherited in a series of generations of rodents, are discussed.     

Accumulation of 90Sr in the bone tissue of the northern mole vole, living in the head part of the East Ural Radioactive Trace

Deposition of 90Sr in a bone tissue of the northern mole-vole (Ellobius talpinus Pallas, 1770) living in the head part of the East Ural Radioactive Trace (EURT) (density of pollution by 90Sr 37 MBq/m2 - 1000 Ci/km2) was investigated. Features of the rodent are digging (underground) way of life, family organization of settlements and weak migratory activity. Authentic (p < 0.01) interfamily distinctions in concentration 90Sr are revealed on the background of absence of sexual and are features of accumulation the radionuclides inside separate families, thus individual parameters differ in 7 times. It is established, that the level of accumilation of 90Sr is comparable with that in other species of small mammals in this territory. Age inversion of accumulation of 90Sr earlier found out in other species of mammals on the EURT is confirmed.     

Functional morphology of the ear in fossorial rodents, Microtus arvalis and Arvicola terrestris

Functionally relevant features and parameters of the outer, middle, and inner ear were studied morphologically and morphometrically in two species of voles, smaller Microtus arvalis and larger Arvicola terrestris. The findings in these fossorial (i.e., burrowing) rodents with components of surface activity were compared with respective findings reported for taxonomically related muroid rodents representing the same size classes but different eco-morphotypes: obligate subterranean rodents (Ellobius talpinus and Spalax ehrenbergi superspecies) and generalized rodents (Mus domesticus and Rattus norvegicus). The ear in voles was characterized by traits reported for subterranean rodents. The eardrum was round, without a distinct pars flaccida, and had an area of 5.4 mm2 in M. arvalis and 9 mm2 in A. terrestris. The middle ear exhibited reduced goniale, enlarged incus nearly parallel to the manubrium of the malleus. The malleus-incus lever ratio amounted to 2.1 (M. arvalis) and 2.0 (A. terrestris). The malleus-incus complex weighed about 0.8 mg in both vole species. The stapedial footplate had an area of 0.3 mm2 in M. arvalis and 0.4 mm2 in A. terrestris. The cochlea had 2.3 coils in both vole species; the basilar membrane was 8.5 mm and 10.5 mm long in M. arvalis and A. terrestris, respectively. There were on average 1,030 (M. arvalis) and 1,220 (A. terrestris) inner hair cells, and 3,760 (M. arvalis) and 4,250 (A. terrestris) outer hair cells in the organ of Corti. In quantitative terms, all these (as well as some further) traits and parameters were intermediate (related to body size) between those reported for generalized rodents on the one hand and subterranean ones on the other. The sound transmission system of the ear seems to be best tuned to frequencies of about 8-16 kHz with a high-frequency cut-off at about 50-60 kHz. The ear of A. terrestris seems to be tuned to somewhat lower frequencies than that in M. arvalis. In this aspect as well as regarding hearing sensitivity (as judged from the mechanical transmission parameters), voles can be considered intermediate not only in their lifestyle but also in their hearing abilities between the subterranean rodents (mole-vole and blind mole-rat) and the surface dwellers (house mouse and Norway rat).     

The characteristics of 90Sr accumulation and elimination in residents of the Urals region in the period of 1957-1988

A number of radiation accidents occurred at the "Mayak" plutonium production complex (Southern Urals, Russia) in 1950-60s that resulted in environmental contamination of large territories in the Southern and Middle Urals with long-lived radionuclides (90Sr, 137Cs). The results of long-term radiological monitoring were compiled into special databases at the Urals Research Center for Radiation Medicine, which allow the study of 90Sr behaviour in the environment and its accumulation in the human body. Retrospective analysis of the data on postmortem measurements of 90Sr in bones performed on residents of the Urals region over 1957-1988 were conducted for the first time. A time-dependence of 90Sr accumulation in the human body upon its density in soils was observed. Age and gender characteristics of 90Sr retention in bones were studied. It was shown that the levels of 90Sr in residents of large territories north and north-east of Chelyabinsk region, and of bordering territories of Sverdlovsk region were several times higher than the average values for the Russian Federation. Based on the measurements the direct dependence of 90Sr content in the skeleton on 90Sr density in soils was observed 30 years after the 1957 accident (the so called "Kyshtym accident"). However, for individual settlements this dependence may be weakened as a result of the influence of different biological and social factors. No influence of gender on 90Sr accumulation in the skeleton was found in the study. The analysis of age characteristics of 90Sr accumulation in bone, as influenced by skeleton growth and maturation, showed maximal levels of 90Sr from global fallout to be accumulated in bones of the persons born in 1949-1955.     

鼹形田鼠入侵鼠和留居鼠之间的相互作用

我们研究了营掘地生活的社会性啮齿类——鼹形田鼠(Ellobius talpinus Pall．)对陌生鼠入侵完整家庭所占据洞穴系统后的行为和内分泌反应。在繁殖期和非繁殖期,所有的入侵鼠都在进入洞穴2天内从留居鼠的洞穴系统内消失;而在引入空洞穴的7只鼹形田鼠中,有4只至少在该洞穴中停留了2天。引入陌生鼠导致留居鼠在释放地的集中,并引起留居鼠和入侵鼠血浆中皮质酮水平的增加。在繁殖期,在引入陌生鼠的当天,也伴随着留居鼠血浆中睾酮增加的现象。所以,对陌生鼠入侵的模拟表明了留居鼹形田鼠有效的社会性防范,这可能是该物种家庭大小和结构稳定的一个重要机制。留居鼠和入侵鼠的相遇激活了压力的生理机制,特别是在繁殖季节更是如此[动物学报50(1)：19-26,2004]。     

Parameters of peroxide oxidation of lipids and the system of its regulation in the tissues of Ellobius talpinus Pall,living on the territory of the East Urals Radioactive Trace

The peroxide oxidation of lipids and the systems of its regulation was tested in Ellobius talpinus Pall, living on the territory of the East Urals Radioactive Trace with the 90Sr contamination density of 1000 Ci/km2. The degree of sensitivity of the examined tissues and organs was revealed, the most considerable parameters were defined for marking radiosensitivity of the taken species, as well as the complex of parameters, allowing classifying an animal as an inhabitant of the clean or the contaminated territory. The found changes are considered as compensatory mechanisms for forming of nonspecific resistance.     

Assessment of radiation exposure of murine rodents at the EURT territories

The study provides data on contemporary levels of radiation exposure of organs and tissues of murine rodents (several species of mice and voles) inhabiting the East-Ural Radioactive Trace. The estimation procedure involves the most advanced approach based on application of appropriate voxel phantom and biokinetic model. Input data for dose assessment are the results of measurements of skeletal ⁹⁰Sr activity concentration. Maximal internal dose to skeleton, accumulated during 45 days, is 303 mGy. Median internal dose rates on the last day before trapping were 0.83, 0.092 and 0.023 mGy/day for animals trapped at the sites with initial (1957) ⁹⁰Sr surface contamination >37 MBq/m², 18.5–37 MBq/m² and 0.074–18.5 MBq/m² respectively. Taking to account internal and external exposures, upper boundary of the ICRP Derived Consideration Reference Level (DCRL) is exceeded on the territory with maximal level of the initial ⁹⁰Sr surface contamination. On the territory with 18.5–37 MBq/m², whole body mean dose rates to murine rodents exceed the lower boundary of DCRL. On the areas with lower level of surface contamination, even the 90-th percentile of dose rate is below the DCRL.     

Frugal strategy as a base of mole-vole (Ellobius talpinus: Rodentia) adaptations to the fossorial way of life

Interspecies comparison of rodents (Rodentia, Cricetidae) of different ecological specialization has revealed dramatic differences in the most essential physiological life history traits between social fossorial mole vole (Ellobius talpinus) and terrestrial rodents. dwarf hamsters (Phodopus) and voles (Clethrionomys). Mole voles have decreased standard and maximal metabolic rates, reduced ability to maintain constant body temperature. lower rates of growth and sexual maturation, lower reproductive output, declined immune and stress response. These life history traits, shaped by the unique peculiarities of the soil environment, provide effective resource re-allocation towards the costly burrowing activity. Spatial and temporal variability of ecological conditions inevitable for any species additionally participate in re-allocation of internal resources. In cold seasons of the year mole voles increase their metabolic activity and become homeothermic. In the north-eastern periphery of the species range, mole voles are fare more stressed and differ from the individuals inhabit the central part of the range with decreased longevity and increased reproductive output on the level of the entire population.     

Factors of maintaining chromosome polymorphism in common vole Microtus arvalis Pallas, 1779: reduced fertility and meiotic drive

The common vole Microtus arvalis (the form obscurus) exhibits polymorphism of a pericentric inversion in chromosome pair 5 throughout the species range. In the Urals populations, the frequency of an acrocentric variant of the heteromorphic chromosome is very low (on average 3.2%) and virtually does not change annually. The factors of maintaining stable chromosomal polymorphism in the common vole were studied under conditions of a laboratory colony. Heterozygous and homozygous for the acrocentric chromosome females showed a significant reduction of the reproductive output irrespective of the male karyotype. This effect was manifested mostly in litter size at birth. A number of cytogenetic and exophenotypic characteristics, as well as parent--offspring transmission of this chromosome in crosses of various types, were examined. We have found meiotic drive in favor of the acrocentric, as a result of which the frequency of the acrocentric (without taking into account the postnatal mortality) totaled over all cross variants (0.48) was significantly higher than that expected with random segregation (0.42). It is likely that meiotic drive of the acrocentric largely compensates for the reduced fertility of its carriers, being among the factors of maintaining it in natural populations.     

Reconstruction of 90Sr intake for breast-fed infants in the Techa riverside settlements

The Techa River (Southern Urals, Russia) was contaminated as a result of radioactive releases by the Mayak plutonium production facility during 1949-1956. The persons born after the onset of the contamination have been identified as the "Techa River Offspring Cohort" (TROC). The TROC has the potential to provide direct data on health effects in progeny that resulted from exposure of a general parent population to chronic radiation. The purpose of the present investigation is the estimation of (90)Sr intake from breast milk and river water in the period from birth to 6 months of life, necessary for an infant dose calculation. The investigation is based on all available data concerning radioactive contamination due to global fallouts and Mayak releases in the Southern Urals where extensive radiometric and radiochemical investigations of human tissues and environmental samples were conducted during the second half of the twentieth century. The strontium transfer factor from mother's daily diet to breast milk was estimated as 0.05 (0.01-0.13) d L(-1). Based on this transfer factor and data on (90)Sr water contamination, the average total (90)Sr intake for an infant born in the middle Techa River region was found to be equal to 60-80 kBq in 1950-1951. For the same period, calculations of (90)Sr intake using ICRP models gave values of 70-100 kBq. From 1952 onwards, the differences in intakes calculated using the two approaches increased, reaching a factor of 2-3 in 1953. The Techa River data provide the basis for improving and adapting the ICRP models for application to Techa River-specific population.     

A comparative analysis of the mole vole sibling species Ellobius tancrei and E. talpinus (Cricetidae, Rodentia) through chromosome painting and examination of synaptonemal complex structures in hybrids

A comparative genomic analysis was carried out in the mole vole sibling species Ellobius tancrei and E. talpinus. Performing fluorescent in situ hybridisation (Zoo-FISH) using chromosome paints from the field vole Microtus agrestis showed no differences in the allocation of syntenic groups in the karyotypes of these sibling species. The only difference between their karyotypes was the position of the centromere in one pair of chromosomes, which is assumed to be the result of an inversion. To verify this hypothesis, we analysed chromosome synapsis in prophase I of meiosis. We utilised a synaptonemal complex (SC) surface-spreading technique to visualise the process of chromosome synapsis in the spermatocytes and oocytes of first-generation hybrids and back-crosses of these sibling species. In prophase I of meiosis, immunocytochemical and electron microscopy analyses revealed that all bivalents had been fully adjusted. Even in the case of a submetacentric-acrocentric bivalent with different centromere locations, synapsis of SC lateral elements was fulfilled along the entire length of the chromosomes and the formation of an inversion loop was not observed. We hypothesise that a possible mechanism leading to the change in centromere position is the repositioning and/or generation of a neocentromere. Despite the great similarity in the karyotypes of these sibling species, they exhibited significant genomic diversification, which manifested as hybrid sterility and parous female death.     

Reconstruction of radionuclide intakes for the residents of East Urals Radioactive Trace (1957–2011)

The East Urals Radioactive Trace (EURT) was formed after a chemical explosion in the radioactive waste-storage facility of the Mayak Production Association in 1957 (Southern Urals, Russia) and resulted in an activity dispersion of 7.4?×?10¹⁶ Bq into the atmosphere. Internal exposure due to ingestion of radionuclides with local foodstuffs was the main factor of public exposure at the EURT. The EURT cohort, combining residents of most contaminated settlements, was formed for epidemiological study at the Urals Research Center for Radiation Medicine, Russia (URCRM). For the purpose of improvement of radionuclide intake estimates for cohort members, the following data sets collected in URCRM were used: (1) Total β-activity and radiochemical measurements of ⁹⁰Sr in local foodstuffs over all of the period of interest (1958–2011; n?=?2200), which were used for relative ⁹⁰Sr intake estimations. (2) ⁹⁰Sr measurements in human bones and whole body (n?=?338); these data were used for average ⁹⁰Sr intake derivations using an age- and gender-dependent Sr-biokinetic model. Non-strontium radionuclide intakes were evaluated on the basis of ⁹⁰Sr intake data and the radionuclide composition of contaminated foodstuffs. Validation of radionuclide intakes during the first years after the accident was first carried out using measurements of the feces β-activity of EURT residents (n?=?148). The comparison of experimental and reconstructed values of feces β-activity shows good agreement. ⁹⁰Sr intakes for residents of settlements evacuated 7–14 days after the accident were also obtained from ⁹⁰Sr measurements in human bone and whole body. The results of radionuclide intake reconstruction will be used to estimate the internal doses for the members of the EURT cohort.     

Relationship between the study of DNA reassociation kinetics of various chromosomal forms of Ellobius and questions concerning the pathways of chromosome rearrangement during evolution

Fifteen chromosome forms of Ellobius talpinus (from 2n = 31 to 2n = 54) were found in the small area in the Pamirs. Low-chromosome karyotypes evolved from 54-chromosomal ancestral form by Robertsonia centric fusions. The DNA reassociation kinetics of 34- and 54-chromosome forms of E. talpinus have been studied. For comparison DNA of E. lutescens (2n = 17) the karyotype of which seems to have arisen from 54-chromosome ancestor by Robertsonian and other types rearrangements was examined. Reassociation profiles of Ellobius DNA suggest the existence of several repeated sequences families with different frequences of repetitions. The reassociation curves of DNA from 34- and 54-chromosome forms were identical. These data indicate absence of changes in DNA molecular organization during the evolution of E. talpinus karyotypes by Robertsonian fusions. Comparative analysis of DNA reassociation kinetics of E. talpinus and E. lutescens showed identical characteristics of highly repeated sequences and of one from the three intermediate fractions, however Cot 1/2, complexity and repetitive frequencies of two intermediate fractions of E. talpinus and E. lutescens were different. It is possible that non-robertsonian rearrangements of E. lutescens karyotype affected only intermediate repetitions. The alternative explanation of these data is a simple divergence of repeated sequences during the evolution of E. lutescens DNA.     

Gene for structural proteins of the SMC family in the common vole Microtus arvalis]

Genes for four subfamilies of SMC (structural maintenance of chromosomes) proteins have been isolated from the genome of a common vole Microtus arvalis. The high degree of homology between representatives of each SMC protein subfamily of different classes of organisms has been demonstrated. The full-sized copy of a mammalian gene encoding SMC4 protein has been isolated and analyzed for the first time. The SMC proteins enter into the composition of complexes responsible for cohesion of sister chromatids, formation of mitotic chromosomes, recombination, DNA repair, and regulation of gene expression. We discuss the possible participation of the SMC proteins in inactivation of the X chromosome in mammalian females. Common voles of genus Microtus group "arvalis" serve a unique model for the study of the inactivation process.     

Chronic low-dose exposure in the Techa River Cohort: risk of mortality from circulatory diseases

The aim of the present study was to analyze the mortality from circulatory diseases for about 30,000 members of the Techa River cohort over the period 1950–2003, and to investigate how these rates depend on radiation doses. This population received both external and internal exposures from ⁹⁰Sr, ⁸⁹Sr, ¹³⁷Cs, and other uranium fission products as a result of waterborne releases from the Mayak nuclear facility in the Southern Urals region of the Russian Federation. The analysis included individualized estimates of the total (external plus internal) absorbed dose in muscle calculated based on the Techa River Dosimetry System 2009. The cohort-average dose to muscle tissue was 35 mGy, and the maximum dose was 510 mGy. Between 1950 and 2003, 7,595 deaths from circulatory diseases were registered among cohort members with 901,563 person years at risk. Mortality rates in the cohort were analyzed using a simple parametric excess relative risk (ERR) model. For all circulatory diseases, the estimated excess relative risk per 100 mGy with a 15-year lag period was 3.6 % with a 95 % confidence interval of 0.2–7.5 %, and for ischemic heart disease it was 5.6 % with a 95 % confidence interval of 0.1–11.9 %. A linear ERR model provided the best fit. Analyses with a lag period shorter than 15 years from the beginning of exposure did not reveal any significant risk of mortality from either all circulatory diseases or ischemic heart disease. There was no evidence of an increased mortality risk from cerebrovascular disease (p > 0.5). These results should be regarded as preliminary, since they will be updated after adjustment for smoking and alcohol consumption.     

Fish otoliths as biological dosimeter: internal dose calculation

Otoliths are organs used by fish for hearing and keeping balance. They consist of biogenic crystals of hydroxyapatite and do not contain any living cells. Upon exposure to ionizing radiation, otolith hydroxyapatite accumulates radiation-induced stable CO₂^? radicals whose amount is proportional to absorbed dose. In electron paramagnetic resonance (EPR) dosimetry, carbonate ions are registered and, hence, the total accumulated dose in the fish otolith can be quantified. Therefore, otoliths can be used as individual fish dosimeters to support radiobiological and radioecological studies. An important aspect of otolith-based EPR dosimetry on fish from contaminated water bodies is the potential presence of bone-seeking ⁹⁰Sr. Consequently, cumulative absorbed doses measured with EPR in otoliths may reflect the superposition of internal exposure to ⁹⁰Sr/⁹⁰Y and external exposure due to radionuclides circulating in soft tissue of the fish as well as due to environmental contamination. The objective of the present study was to develop a method that allows for an assessment of the contribution of ⁹⁰Sr to the total dose in otolith. The method has been tested using otoliths from seven fish taken from reservoirs located in the Southern Urals contaminated with radionuclides including ⁹⁰Sr. It has been shown that dose to otoliths is largely determined by ⁹⁰Sr in the hydroxyapatite. The internal dose component can be calculated using activity concentration-to-dose conversion factors, which vary slightly in the range of 2.0–2.8?×?10^–3 Gy year^?1 per Bq g^?1 depending on fish species and age. Internal doses to fish from water bodies with different levels of ⁹⁰Sr contamination were calculated in the range from 2 mGy to?~?200 Gy. External dose contribution was derived for two fish only to be about 100 and 40 Gy. It is concluded that EPR dosimetry on fish otoliths is a promising tool when external exposure prevails or is comparable to internal exposure due to ⁹⁰Sr.

     

Influence of chronic exposure to low doses of γ-radiation and <Superscript>90</Superscript>Sr on the level of DNA breaks and cell sensitivity to hydrogen peroxide in the mouse spleen

Using comet assay, a statistically significant increase (p < 0.05) in the level of DNA breaks in spleen cells was revealed in male CBA/lac mice exposed to γ-radiation (1.7 mGy/day) or ⁹⁰Sr (150–250 Bq/day) for 210 days. The level of DNA breaks also increased under combined exposure to both γ-radiation and ⁹⁰Sr (p < 0.05), but to a lesser degree than under exposure to each of these factors alone. Upon additional in vitro treatment of spleen cells with hydrogen peroxide, the relative increase in the level of DNA breaks was smaller in cells of irradiated mice than in the control. The ratio of the level of DNA breaks after hydrogen peroxide treatment to that before this treatment in control mice was 4.2 ± 0.9, compared to 1.4 ± 0.6 in γ-irradiated mice, 1.9 ± 0.8 in ⁹⁰Sr-irradiated mice, and 2.3 ± 0.8 in mice exposed to both γ- and ⁹⁰Sr-irradiation.     

The estimation of 90Sr and 137Cs excretion parameters from the organism of wild and laboratory small mammals in vivo, after natural contamination of them in the Chernobyl zone

In the course of laboratory experiment, parameters of 90Sr and 137Cs excretion were estimated in individuals of bank vole Clethrionomys glareolus, captured at one of the most contaminated sites of the Chernobyl zone. The animals were kept under laboratory conditions using "clean" feed during 50 days. The similar investigation was carried out with laboratory mice Mus musculus (Big Blue line) during a longer period (184 days). The measurements of 90Sr content in the animals' body were in vivo carried out, using a specially designed beta-spectrometer with appropriate software, and 137Cs one--by the gamma-spectrometer. During the experiment, the animals had lost less 0.4% of activity due to physical decay of radionuclides. The organism was depurated mainly through biological excretion. In accordance with parameters of one-component exponential decay equation, 99.3% of 137Cs initial content in vole was excreted with half-life period of 2.18 days, and mice--4.4 days (99%). 90Sr excretion had longer half-life period: 11.7 days (56%) in voles, and 49.9 days (87%) in laboratory mice. The rest radionuclides amount of given model was considered as non-excreted from the organism during the observation period. It was determined on the example of voles that 90Sr and 137Cs loss in males was faster than in females, and among females more intensive excretion was in lactating females. 137Cs excretion from the body of bank vole is mainly with urine (74.7%), whereas 90Sr one--with feces and urine in approximately equal amounts. Due to the birth of babies and consequent feeding female lose appreciably less amount of radionuclides body burden than at daily loss with urine and feces.