- Email: [email protected]
59. Current status of tundra cranes species populations in Yakutia
358
Abstracts / Cryobiology 65 (2012) 339–366
a gap between freezing and melting point, as well as ice re-crystallization inhibition activity. Thermal hysteresis activity of the FfIBP was approximately 2.5 °C at 50 lM that is 10 times higher than moderately active LeIBP. Furthermore, the ice re-crystallization inhibition activity represented by the FfIBP has the ability to inhibit ice growing at low concentration 2.5 lM limit. Consequently the FfIBP was classified as a hyperactive ice binding protein. Also, preliminary X-ray crystallography was performed to reveal the ice-binding site of FfIBP at the molecular level. Conflict of interest: None declared. Acknowledgement We thank the staff at the X-ray core facility of Korea Basic Science Institute (KBSI) (Ochang, Korea). This work was supported by the National Agenda Project from the Korea Research Council of Fundamental Science and Technology (KRCF) (Grant No. PG11010 to HJK) and the Korea Polar Research Institute (KOPRI) (PE11100 to HJK and PE11120 to JHL). E-mail address: [email protected] (H.J. Kim) http://dx.doi.org/10.1016/j.cryobiol.2012.07.058
58. Ecological–physiological adaptations of terrestrial vertebrate species to the conditions of sharply continental climate of Yakutia. N.G. Solomonov, E.S. Solomonov, A.I. Anufriev, I.M. Okhlopkov, A.P. Isaev, T.N. Solomonova, V.T. Sedalishchev, N.I. Mordosova, Institute of Biological Problems of Cryolithozone SD RAS, Yakutsk, Russia NorthEastern Federal University, Yakutsk, Russia Middendorf (1869) raised the question of the main ways of adaptation of Siberian animals to the cold climate. He paid attention to the fact that northern mammals have thick and mossy fur, accumulate more fat for winter and have such a lifestyle that allows saving warmth in the body. Subsequent research has shown that the basis of the processes of adaptation of animals to the extreme conditions is made by the preservation of their energy balance in the process of interaction with the environment and preservation of relative stability of internal environment of the organism. Since the 1960s we have conducted research of adaptations of terrestrial vertebrate species to the cold climate conditions in the taiga and tundra zones of Yakutia. The main direction of our research is studying morphological–physiological and ecological parameters of the species securing the preservation of their energy balance in the harsh conditions of North-East Asia. The main objects of research are mass species of mammals, birds and amphibian. We have studied auto ecology, population ecology and ecological physiology of the model species. Special attention was paid to the ecological peculiarities of hibernating and large cold-adapted mammals: reindeer, elk, Yakut-horse, brown bear. It was found that during preparation to winter these organisms produce biologically active substances, which reduces the level of metabolism in winter, similar to those of hibernating species in the condition of hibernation. Economy of energy resources of widespread rodents is provided both by reducing the level of metabolism within the thermo-neutral zone and better development of physical thermoregulation of aboriginal forms, and, especially, by adaptive behavior. For mammals’ and birds’ ability to adapt to the cold, their morphological peculiarities of fur and feathers, composition of their lower limbs, anatomic peculiarities of nasal cavity and its blood supply are of great significance, also their adaptive behavior. The strategy of avoiding cold of two amphibian species of Yakutia – Siberian wood frog Rana amurensis and Asiatic salamander Salamandrella keyserlingii – is implemented differently: frogs winter in underwater pits lapsing into hypobiosis and salamanders find convenient wintering places on the land, sometimes far from reservoirs, i.e. frogs winter at around zero temperatures, whereas salamanders, who winter almost under a bedding, experience winter temperatures of the soil surface level to minus 30– 32 °C. In these conditions the strategy of avoiding cold does not help and there has to be another strategy of cold (frost) tolerance, which can be secured by producing glycerol and other chemical agents in the organism. Crow birds feeding with more high-calorie food in winter sustain a high level of metabolism, whereas gallinaceous, whose food is less high-calorie, have to use a strategy of cold avoiding by keeping warm under snow shelters. Aboriginal forms of mammals (reindeer, Yakut-horse, musk-ox) are typical of lower level metabolism and body temperature in winter months. There are substantial differences in using tundra by the reindeer and musk-ox. The first one makes long migrations to winter pastures to forest tundra and summer tundra habitats, the musk-ox one has more settled lifestyle. Conflict of interest: None declared. Source of funding: None declared. E-mail address: [email protected] (N.G. Solomonov) http://dx.doi.org/10.1016/j.cryobiol.2012.07.059
59. Current status of tundra cranes species populations in Yakutia. N.I. Germogenov, I.P. Bysykatova, A.G. Degtyarev, N.G. Solomonov, S.M. Sleptsov, N.N. Egorov, A.E. Pshennikov, M.V. Vladimirtseva, Institute for Biological Problems of Cryolithozone, Siberian Branch of Russian Academy for Sciences, Lenina Ave. 41, 677980 Yakutsk, Russia Just two from four breeding species of cranes in Yakutia, Siberian Crane Grus leucogeranus and Sandhill crane Grus canadensis canadensis, are found in the most northern areas of the region, inhabiting the subarctic tundra of the lowland between rivers Yana and Kolyma. Siberian crane is represented by the eastern population in the region. This crane winters in China (Yangtze River downstream basin) and appears to be a typical species of the Siberian fauna and endangered bird of the world fauna. Sandhill crane is an expanding species of American fauna with wintering grounds in Texas (USA) and Mexico. This crane is in a process of active distribution in the northeastern Yakutia and related to rare endangered species of the region. Siberian cranes’ increase of the eastern population number has been observed since the late 1990s on the breeding ground, particularly in tundra of the River Indigirka basin (the number of crane pairs increased from 52 to 56 on the model site of 1314 km2) and on the main wintering ground in Poyang Lake Nature Reserve in the southeast of China (it reached 4 thousand individuals), no doubt, was related to the active scientific and conservation work undertaken within the framework of international cooperation, with a reduction of disturbance factors in key areas of habitat. Nevertheless, the current status of the population is still estimated to be alarming. The optimal habitat of Siberian Cranes is lakeside lowlands and wetlands occupied by 21–26% of the breeding range. They are located outside of the regular spring flooding territories and in uneven and mosaic distribution. The birds inhabit mainly areas adjacent to large and medium-sized lakes, occupying no more than 50–60% of the areas suitable for their nesting. The water content increases in the territory because of the destruction and defrosting of tundra soils that leads to a reduction of the most optimum habitat for the population. An average home range size for a reproductive pair, including its breeding and feeding areas, is 7.3–16.5 or 10.6 km2. The rate of population growth (reproduction efficiency varies from 4.3–8.7 to 65–83.3%) allows us to assume that the absence of separate generations of birds which did not live to maturity was due to their very low number and natural losses. The front of the bird migration in Yakutia has a quite narrow corridor. This is especially noticeable in the middle Aldan River at fall where up to 50% of the eastern population is reported from the separate observation points. Siberian crane transit stops are much longer in China than in Russia, and the concentrations of birds are more numerous in China. Sandhill cranes continue to distribute in the western region of tundra and increase their numbers. At present, the range of this species almost completely coincides with that of the Siberian crane (approximately 82 km2). The species number has grown tenfold and reached 56 pairs over the past 20 years on the model site in tundra of the River Indigirka basin. Conflict of interest: None declared. Source of funding: None declared. E-mail addresses: [email protected], [email protected] (N.I. Germogenov) http://dx.doi.org/10.1016/j.cryobiol.2012.07.060
60. Investigation of osmotic tolerance limits for rational design of vitrification procedures. Allyson K. Fry, Adam Z. Higgins, School of Chemical, Biological and Environmental Engineering, Oregon State University, United States Previously, we reported a new strategy for mathematical optimization of CPA addition and removal procedures which involves minimization of a toxicity cost function while constraining cell volumes within osmotic tolerance limits. The purpose of this study was to clearly define cellular osmotic tolerance limits of bovine pulmonary artery endothelial cells for inclusion in our optimization routine. Our general experimental approach consisted of exposure to anisotonic solutions for a prescribed time period, return to isotonic solution, and measurement of cell viability. The variables investigated include the buffering system, the type of non-permeating solute, the duration of exposure to anisotonic solutions, and the temperature. Primary bovine endothelial cells cultured in 96 well plates were exposed to solutions with concentrations ranging from pure water to 4000 mOsm/kg. Viability was assessed directly before and 24 h after treating with test solutions using the resazurin-based PrestoBlue assay. The resulting hypotonic and hypertonic viability trends were fit with 3-parameter logistic models to facilitate identification of osmotic tolerance limits. The best-fit logistic models were used to determine the hypoand hypertonic concentrations that resulted in 90% cell yield relative to untreated cells. We first investigated the effect of the buffering system on sensitivity to osmotic damage. Cells retained greater viability after 15 min exposure to anisotonic solutions that were buffered with HEPES rather than phosphate. Thus, subsequent experiments were performed using HEPES buffered test solutions. We next investigated the effects of the type of non-permeating solute and the temperature on sensitivity to osmotic damage. Hypertonic solutions composed of NaCl were less
Abstracts / Cryobiology 65 (2012) 339–366
a gap between freezing and melting point, as well as ice re-crystallization inhibition activity. Thermal hysteresis activity of the FfIBP was approximately 2.5 °C at 50 lM that is 10 times higher than moderately active LeIBP. Furthermore, the ice re-crystallization inhibition activity represented by the FfIBP has the ability to inhibit ice growing at low concentration 2.5 lM limit. Consequently the FfIBP was classified as a hyperactive ice binding protein. Also, preliminary X-ray crystallography was performed to reveal the ice-binding site of FfIBP at the molecular level. Conflict of interest: None declared. Acknowledgement We thank the staff at the X-ray core facility of Korea Basic Science Institute (KBSI) (Ochang, Korea). This work was supported by the National Agenda Project from the Korea Research Council of Fundamental Science and Technology (KRCF) (Grant No. PG11010 to HJK) and the Korea Polar Research Institute (KOPRI) (PE11100 to HJK and PE11120 to JHL). E-mail address: [email protected] (H.J. Kim) http://dx.doi.org/10.1016/j.cryobiol.2012.07.058
58. Ecological–physiological adaptations of terrestrial vertebrate species to the conditions of sharply continental climate of Yakutia. N.G. Solomonov, E.S. Solomonov, A.I. Anufriev, I.M. Okhlopkov, A.P. Isaev, T.N. Solomonova, V.T. Sedalishchev, N.I. Mordosova, Institute of Biological Problems of Cryolithozone SD RAS, Yakutsk, Russia NorthEastern Federal University, Yakutsk, Russia Middendorf (1869) raised the question of the main ways of adaptation of Siberian animals to the cold climate. He paid attention to the fact that northern mammals have thick and mossy fur, accumulate more fat for winter and have such a lifestyle that allows saving warmth in the body. Subsequent research has shown that the basis of the processes of adaptation of animals to the extreme conditions is made by the preservation of their energy balance in the process of interaction with the environment and preservation of relative stability of internal environment of the organism. Since the 1960s we have conducted research of adaptations of terrestrial vertebrate species to the cold climate conditions in the taiga and tundra zones of Yakutia. The main direction of our research is studying morphological–physiological and ecological parameters of the species securing the preservation of their energy balance in the harsh conditions of North-East Asia. The main objects of research are mass species of mammals, birds and amphibian. We have studied auto ecology, population ecology and ecological physiology of the model species. Special attention was paid to the ecological peculiarities of hibernating and large cold-adapted mammals: reindeer, elk, Yakut-horse, brown bear. It was found that during preparation to winter these organisms produce biologically active substances, which reduces the level of metabolism in winter, similar to those of hibernating species in the condition of hibernation. Economy of energy resources of widespread rodents is provided both by reducing the level of metabolism within the thermo-neutral zone and better development of physical thermoregulation of aboriginal forms, and, especially, by adaptive behavior. For mammals’ and birds’ ability to adapt to the cold, their morphological peculiarities of fur and feathers, composition of their lower limbs, anatomic peculiarities of nasal cavity and its blood supply are of great significance, also their adaptive behavior. The strategy of avoiding cold of two amphibian species of Yakutia – Siberian wood frog Rana amurensis and Asiatic salamander Salamandrella keyserlingii – is implemented differently: frogs winter in underwater pits lapsing into hypobiosis and salamanders find convenient wintering places on the land, sometimes far from reservoirs, i.e. frogs winter at around zero temperatures, whereas salamanders, who winter almost under a bedding, experience winter temperatures of the soil surface level to minus 30– 32 °C. In these conditions the strategy of avoiding cold does not help and there has to be another strategy of cold (frost) tolerance, which can be secured by producing glycerol and other chemical agents in the organism. Crow birds feeding with more high-calorie food in winter sustain a high level of metabolism, whereas gallinaceous, whose food is less high-calorie, have to use a strategy of cold avoiding by keeping warm under snow shelters. Aboriginal forms of mammals (reindeer, Yakut-horse, musk-ox) are typical of lower level metabolism and body temperature in winter months. There are substantial differences in using tundra by the reindeer and musk-ox. The first one makes long migrations to winter pastures to forest tundra and summer tundra habitats, the musk-ox one has more settled lifestyle. Conflict of interest: None declared. Source of funding: None declared. E-mail address: [email protected] (N.G. Solomonov) http://dx.doi.org/10.1016/j.cryobiol.2012.07.059
59. Current status of tundra cranes species populations in Yakutia. N.I. Germogenov, I.P. Bysykatova, A.G. Degtyarev, N.G. Solomonov, S.M. Sleptsov, N.N. Egorov, A.E. Pshennikov, M.V. Vladimirtseva, Institute for Biological Problems of Cryolithozone, Siberian Branch of Russian Academy for Sciences, Lenina Ave. 41, 677980 Yakutsk, Russia Just two from four breeding species of cranes in Yakutia, Siberian Crane Grus leucogeranus and Sandhill crane Grus canadensis canadensis, are found in the most northern areas of the region, inhabiting the subarctic tundra of the lowland between rivers Yana and Kolyma. Siberian crane is represented by the eastern population in the region. This crane winters in China (Yangtze River downstream basin) and appears to be a typical species of the Siberian fauna and endangered bird of the world fauna. Sandhill crane is an expanding species of American fauna with wintering grounds in Texas (USA) and Mexico. This crane is in a process of active distribution in the northeastern Yakutia and related to rare endangered species of the region. Siberian cranes’ increase of the eastern population number has been observed since the late 1990s on the breeding ground, particularly in tundra of the River Indigirka basin (the number of crane pairs increased from 52 to 56 on the model site of 1314 km2) and on the main wintering ground in Poyang Lake Nature Reserve in the southeast of China (it reached 4 thousand individuals), no doubt, was related to the active scientific and conservation work undertaken within the framework of international cooperation, with a reduction of disturbance factors in key areas of habitat. Nevertheless, the current status of the population is still estimated to be alarming. The optimal habitat of Siberian Cranes is lakeside lowlands and wetlands occupied by 21–26% of the breeding range. They are located outside of the regular spring flooding territories and in uneven and mosaic distribution. The birds inhabit mainly areas adjacent to large and medium-sized lakes, occupying no more than 50–60% of the areas suitable for their nesting. The water content increases in the territory because of the destruction and defrosting of tundra soils that leads to a reduction of the most optimum habitat for the population. An average home range size for a reproductive pair, including its breeding and feeding areas, is 7.3–16.5 or 10.6 km2. The rate of population growth (reproduction efficiency varies from 4.3–8.7 to 65–83.3%) allows us to assume that the absence of separate generations of birds which did not live to maturity was due to their very low number and natural losses. The front of the bird migration in Yakutia has a quite narrow corridor. This is especially noticeable in the middle Aldan River at fall where up to 50% of the eastern population is reported from the separate observation points. Siberian crane transit stops are much longer in China than in Russia, and the concentrations of birds are more numerous in China. Sandhill cranes continue to distribute in the western region of tundra and increase their numbers. At present, the range of this species almost completely coincides with that of the Siberian crane (approximately 82 km2). The species number has grown tenfold and reached 56 pairs over the past 20 years on the model site in tundra of the River Indigirka basin. Conflict of interest: None declared. Source of funding: None declared. E-mail addresses: [email protected], [email protected] (N.I. Germogenov) http://dx.doi.org/10.1016/j.cryobiol.2012.07.060
60. Investigation of osmotic tolerance limits for rational design of vitrification procedures. Allyson K. Fry, Adam Z. Higgins, School of Chemical, Biological and Environmental Engineering, Oregon State University, United States Previously, we reported a new strategy for mathematical optimization of CPA addition and removal procedures which involves minimization of a toxicity cost function while constraining cell volumes within osmotic tolerance limits. The purpose of this study was to clearly define cellular osmotic tolerance limits of bovine pulmonary artery endothelial cells for inclusion in our optimization routine. Our general experimental approach consisted of exposure to anisotonic solutions for a prescribed time period, return to isotonic solution, and measurement of cell viability. The variables investigated include the buffering system, the type of non-permeating solute, the duration of exposure to anisotonic solutions, and the temperature. Primary bovine endothelial cells cultured in 96 well plates were exposed to solutions with concentrations ranging from pure water to 4000 mOsm/kg. Viability was assessed directly before and 24 h after treating with test solutions using the resazurin-based PrestoBlue assay. The resulting hypotonic and hypertonic viability trends were fit with 3-parameter logistic models to facilitate identification of osmotic tolerance limits. The best-fit logistic models were used to determine the hypoand hypertonic concentrations that resulted in 90% cell yield relative to untreated cells. We first investigated the effect of the buffering system on sensitivity to osmotic damage. Cells retained greater viability after 15 min exposure to anisotonic solutions that were buffered with HEPES rather than phosphate. Thus, subsequent experiments were performed using HEPES buffered test solutions. We next investigated the effects of the type of non-permeating solute and the temperature on sensitivity to osmotic damage. Hypertonic solutions composed of NaCl were less