Vitrification of one-cell mouse embryos in cryotubes

Preventing intracellular ice formation is essential to cryopreserve cells. Prevention can be achieved by converting cell water into a non-crystalline glass, that is, to vitrify. The prevailing belief is that to achieve vitrification, cells must be suspended in a solution containing a high concentration of glass-inducing solutes and cooled rapidly. In this study, we vitrified 1-cell mouse embryos and examined the effect of the cooling rate, the warming rate, and the concentration of cryoprotectant on cell survival. Embryos were vitrified in cryotubes. The vitrification solutions used were EFS20, EFS30, and EFS40, which contained ethylene glycol (20, 30 and 40% v/v, respectively), Ficoll (24%, 21%, and 18% w/v, respectively) and sucrose (0.4 0.35, and 0.3 M, respectively). A 5-μl EFS solution suspended with 1-cell embryos was placed in a cryotube. After 2 min in an EFS solution at 23 °C, embryos were vitrified by direct immersion into liquid nitrogen. The sample was warmed at 34 °C/min, 4,600 °C/min and 6,600 °C/min. With EFS40, the survival was low regardless of the warming rate. With EFS30 and EFS20, survival was also low when the warming rate was low, but increased with higher warming rates, likely due to prevention of intracellular ice formation. When 1-cell embryos were vitrified with EFS20 and warmed rapidly, almost all of the embryos developed to blastocysts in vitro. Moreover, when vitrified 1-cell embryos were transferred to recipients at the 2-cell stage, 43% of them developed to term. In conclusion, we developed a vitrification method for 1-cell mouse embryos by rapid warming using cryotubes.     

To aggregate or not? Capturing the spatio-temporal complexity of the thermal regime

Freshwater stream systems are under immense pressure from various anthropogenic impacts, including climate change. Stream systems are increasingly being altered by changes to the magnitude, timing, frequency, and duration of their thermal regimes, which will have profound impacts on the life-history dynamics of resident biota within their home range. Although temperature regimes have a significant influence on the biology of instream fauna, large spatio-temporal temperature datasets are often reduced to a single metric at discrete locations and used to describe the thermal regime of a system; potentially leading to a significant loss of information crucial to stream management. Models are often used to extrapolate these metrics to unsampled locations, but it is unclear whether predicting actual daily temperatures or an aggregated metric of the temperature regime best describes the complexity of the thermal regime. We fit spatial statistical stream-network models (SSNMs), random forest and non-spatial linear models to stream temperature data from the Upper Condamine River in QLD, Australia and used them to semi-continuously predict metrics describing the magnitude, duration, and frequency of the thermal regime through space and time. We compared both daily and aggregated temperature metrics and found that SSNMs always had more predictive ability than the random forest models, but both models outperformed the non-spatial linear model. For metrics describing thermal magnitude and duration, aggregated predictions were most accurate, while metrics describing the frequency of heating events were better represented by metrics based on daily predictions generated using a SSNM. A more comprehensive representation of the spatio-temporal thermal regime allows researchers to explore new spatio-temporally explicit questions about the thermal regime. It also provides the information needed to generate a suite of ecologically meaningful metrics capturing multiple aspects of the thermal regime, which will increase our scientific understanding of how organisms respond to thermal cues and provide much-needed information for more effective management actions.     

Contrasting population changes in sympatric penguin species in association with climate warming

Climate warming and associated sea ice reductions in Antarctica have modified habitat conditions for some species. These include the congeneric Adélie, chinstrap and gentoo penguins, which now demonstrate remarkable population responses to regional warming. However, inconsistencies in the direction of population changes between species at different study sites complicate the understanding of causal processes. Here, we show that at the South Orkney Islands where the three species breed sympatrically, the less ice‐adapted gentoo penguins increased significantly in numbers over the last 26 years, whereas chinstrap and Adélie penguins both declined. These trends occurred in parallel with regional long‐term warming and significant reduction in sea ice extent. Periodical warm events, with teleconnections to the tropical Pacific, caused cycles in sea ice leading to reduced prey biomass, and simultaneous interannual population decreases in the three penguin species. With the loss of sea ice, Adélie penguins were less buffered against the environment, their numbers fluctuated greatly and their population response was strong and linear. Chinstrap penguins, considered to be better adapted to ice‐free conditions, were affected by discrete events of locally increased ice cover, but showed less variable, nonlinear responses to sea ice loss. Gentoo penguins were temporarily affected by negative anomalies in regional sea ice, but persistent sea ice reductions were likely to increase their available niche, which is likely to be substantially segregated from that of their more abundant congeners. Thus, the regional consequences of global climate perturbations on the sea ice phenology affect the marine ecosystem, with repercussions for penguin food supply and competition for resources. Ultimately, variability in penguin populations with warming reflects the local balance between penguin adaptation to ice conditions and trophic‐mediated changes cascading from global climate forcing.     

The sea-skater Halobates (Heteroptera: Gerridae) – probable cause for extinction in the Mediterranean and potential for re-colonisation following climate change

Sea-skaters in the genus Halobates Eschscholtz 1822 include some of the most specialised water striders and are found in tropical and subtropical seas around the world. Even though species of Halobates occur in both the Atlantic Ocean and the Red Sea, no extant sea-skater has been reported from the Mediterranean Sea. A fossil, Halobates ruffoi Andersen et al., 1994, described from Middle–Upper Eocene (45 Ma) Italy indicates that sea skaters were present in this part of the world in the past. Other geological evidence points to dramatic changes in the Mediterranean Sea during the Tertiary and Quaternary that may have led to their later extinction. In this paper we review briefly the distribution, systematics, evolution and ecology of Halobates, and discuss the potential for the Mediterranean to be recolonised following expected environmental changes due to global warming.     

Community and ecosystem responses to recent climate change

There is ample evidence for ecological responses to recent climate change. Most studies to date have concentrated on the effects of climate change on individuals and species, with particular emphasis on the effects on phenology and physiology of organisms as well as changes in the distribution and range shifts of species. However, responses by individual species to climate change are not isolated; they are connected through interactions with others at the same or adjacent trophic levels. Also from this more complex perspective, recent case studies have emphasized evidence on the effects of climate change on biotic interactions and ecosystem services. This review highlights the ‘knowns’ but also ‘unknowns’ resulting from recent climate impact studies and reveals limitations of (linear) extrapolations from recent climate-induced responses of species to expected trends and magnitudes of future climate change. Hence, there is need not only to continue to focus on the impacts of climate change on the actors in ecological networks but also and more intensively to focus on the linkages between them, and to acknowledge that biotic interactions and feedback processes lead to highly complex, nonlinear and sometimes abrupt responses.     

Differing effects of thermal stress on coral fertilization and early embryogenesis in four Indo Pacific species

Coral reefs are expected to be severely impacted by rising seawater temperatures associated with climate change. The fertilization and early embryogenesis of four reef-building coral species representing three Indo-Pacific families were examined in a series of laboratory experiments where temperatures were increased up to 5–6°C at ambient. High levels of fertilization and normal embryogenesis were observed for Favites abdita, Favites chinensis and Mycedium elephantotus at temperatures to 32°C (+5°C) and embryos developed normally until the 5th cell cleavage. Acropora millepora was the only species to be affected by higher temperatures, exhibiting significantly reduced fertilization and a higher frequency of embryonic abnormalities at 32°C (+4°C), and fertilization ceased altogether at 34°C (+6°C). Early cell cleavage rates increased with temperature up to 32°C for all species.     

Recent increase in body size of the American marten Martes americana in Alaska

Using museum specimens, we studied recent changes in skull size of the American marten Martes americana , in continental Alaska. In Alaska, global warming has resulted in milder winters that may contribute to an improved food supply in the wild. In the present study, we tested the hypothesis that body size of the marten had increased during the second half of the 20th century, in response to global warming. We found that skull size, and by implication body size, increased significantly during the second half of the 20th century, possibly due to an improved food supply and/or lower metabolic demands in winter. Improved food availability in winter may result from the improved nutritional conditions for prey, and/or from increased access to prey resulting from a longer snow-free season. Longitude had a significant positive effect on skull size and a significant negative effect on teeth size. In Alaska, the climate is milder along the western coast and becomes harsher inland. Hence, the milder climate was associated with larger body size providing further support for our prediction that body size of the American marten was influenced by food availability and reduced energy expenditure. The negative relationship between longitude and teeth size may indicate a trend towards a larger prey in inland marten populations, but we have no data to support or refute this hypothesis.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 93 , 701–707.     

Increasing ocean temperatures allow tropical fishes to survive overwinter in temperate waters

The southeast coast of Australia is a global hotspot for increasing ocean temperatures due to climate change. The temperate incursion of the East Australian Current (EAC) is increasing, affording increased connectivity with the Great Barrier Reef. The survival of tropically sourced juveniles over the winter is a significant stumbling block to poleward range shifts of marine organisms in this region. Here we examine the dependence of overwintering on winter severity and prewinter recruitment for eight species of juvenile coral reef fishes which are carried into temperate SE Australia (30–37 °S) by the EAC during the austral summer. The probability of persistence was most strongly influenced by average winter temperature and there was no effect of recruitment strength. Long‐term (138 years) data indicate that winter water temperatures throughout this region are increasing at a rate above the global average and predictions indicate a further warming of >2 °C by the end of the century. Rising ocean temperatures are resulting in a higher frequency of winter temperatures above survival thresholds. Current warming trajectories predict 100% of winters will be survivable by at least five of the study species as far south as Sydney (34 °S) by 2080. The implications for range expansions of these and other species of coral reef fish are discussed.