Impact of thermal stress on evolutionary trajectories of pathogen resistance in three-spined stickleback (Gasterosteus aculeatus), supplementary material

Background: Pathogens are a major regulatory force for host populations, especially under stressful conditions. Elevated temperatures may enhance the development of pathogens, increase the number of transmission stages, and can negatively influence host susceptibility depending on host thermal tolerance. As a net result, this can lead to a higher prevalence of epidemics during summer months. These conditions also apply to marine ecosystems, where possible ecological impacts and the population-specific potential for evolutionary responses to changing environments and increasing disease prevalence are, however, less known. Therefore, we investigated the influence of thermal stress on the evolutionary trajectories of disease resistance in three marine populations of three-spined sticklebacks Gasterosteus aculeatus by combining the effects of elevated temperature and infection with a bacterial strain of Vibrio sp. using a common garden experiment. Results: We found that thermal stress had an impact on fish weight and especially on survival after infection after only short periods of thermal acclimation. Environmental stress reduced genetic differentiation (QST) between populations by releasing cryptic within-population variation. While life history traits displayed positive genetic correlations across environments with relatively weak genotype by environment interactions (GxE), environmental stress led to negative genetic correlations across environments in pathogen resistance. This reversal of genetic effects governing resistance is probably attributable to changing environment-dependent virulence mechanisms of the pathogen interacting differently with host genotypes, i.e. GPathogenxGHostxE or (GPathogenxE)x(GHostxE) interactions, rather than to pure host genetic effects, i.e. GHostxE interactions. Conclusion: To cope with climatic changes and the associated increase in pathogen virulence, host species require wide thermal tolerances and pathogen-resistant genotypes. The higher resistance we found for some families at elevated temperatures showed that there is evolutionary potential for resistance to Vibrio sp. in both thermal environments. The negative genetic correlation of pathogen resistance between thermal environments, on the other hand, indicates that adaptation to current conditions can be a weak predictor for performance in changing environments. The observed feedback on selective gradients exerted on life history traits may exacerbate this effect, as it can also modify the response to selection for other vital components of fitness.

Climate sensitivity across marine domains of life: limits to evolutionary adaptation shape species interactions, supplementary material

Organisms in all domains, Archaea, Bacteria, and Eukarya will respond to climate change with differential vulnerabilities resulting in shifts in species distribution, coexistence, and interactions. The identification of unifying principles of organism functioning across all domains would facilitate a cause and effect understanding of such changes and their implications for ecosystem shifts. For example, the functional specialization of all organisms in limited temperature ranges leads us to ask for unifying functional reasons. Organisms also specialize in either anoxic or various oxygen ranges, with animals and plants depending on high oxygen levels. Here, we identify thermal ranges, heat limits of growth, and critically low (hypoxic) oxygen concentrations as proxies of tolerance in a meta-analysis of data available for marine organisms, with special reference to domain-specific limits. For an explanation of the patterns and differences observed, we define and quantify a proxy for organismic complexity across species from all domains. Rising complexity causes heat (and hypoxia) tolerances to decrease from Archaea to Bacteria to uni- and then multicellular Eukarya. Within and across domains, taxon-specific tolerance limits likely reflect ultimate evolutionary limits of its species to acclimatization and adaptation. We hypothesize that rising taxon-specific complexities in structure and function constrain organisms to narrower environmental ranges. Low complexity as in Archaea and some Bacteria provide life options in extreme environments. In the warmest oceans, temperature maxima reach and will surpass the permanent limits to the existence of multicellular animals, plants and unicellular phytoplankter. Smaller, less complex unicellular Eukarya, Bacteria, and Archaea will thus benefit and predominate even more in a future, warmer, and hypoxic ocean.

Using the critical salinity (Scrit) concept to predict invasion potential of the anemone Diadumene lineata in the Baltic Sea

It is widely assumed that the ability of an introduced species to acclimate to local environmental conditions determines its invasion success. The sea anemone Diadumene lineata is a cosmopolitan invader and shows extreme physiological tolerances. It was recently discovered in Kiel Fjord (Western Baltic Sea), although the brackish conditions in this area are physiologically challenging for most marine organisms. This study investigated salinity tolerance in D. lineata specimens from Kiel Fjord in order to assess potential geographical range expansion of the species in the Baltic Sea. In laboratory growth assays, we quantified biomass change and asexual reproduction rates under various salinity regimes (34: North Sea, 24: Kattegat, 14: Kiel Fjord, 7: Baltic Proper). Furthermore, we used 1H-NMR-based metabolomics to analyse intracellular osmolyte dynamics. Within 4 weeks D. lineata exhibited a 5-fold population growth through asexual reproduction at high salinities (34 and 24). Biomass increase under these conditions was significantly higher (69%) than at a salinity of 14. At a salinity of 7, anemones ceased to reproduce asexually, their biomass decreased and metabolic depression was observed. Five main intracellular osmolytes were identified to be regulated in response to salinity change, with osmolyte depletion at a salinity of 7. We postulate that depletion of intracellular osmolytes defines a critical salinity (Scrit) that determines loss of fitness. Our results indicate that D. lineata has the potential to invade the Kattegat and Skagerrak regions with salinity >10. However, salinities of the Baltic Proper (salinity <8) currently seem to constitute a physiological limit for the species.

Late Eocene to Recent deep-sea benthic foraminifera from the Central equatorial Pacific Ocean

Benthic foraminifers were studied in upper Eocene to Recent core-catcher samples from DSDP Sites 573, 574, and 575. The sites are on a north-south transect from the equator to about 05°N at about 133°W, water depth 4300 to 4600 m. At Site 574 additional samples were used to study the Eocene/Oligocene boundary in detail. About 200 specimens were counted per sample. The fauna is highly diverse (about 50 to 70 species per sample) and is of low dominance. The diversity is not related to age or sub-bottom depth. Many species are cosmopolitan and probably have wide environmental tolerances. Fluctuations in frequency of some taxa (e.g., Nuttallides umbonifera, Epistominella exigua, and Uvigerina spp.) cannot be correlated from one site to another. Several common species (e.g. Oridorsalis umbonatus and Globocassidulina subglobosa) range from late Eocene to Recent. First and last appearances are generally difficult to define precisely because many species are rare. For some species these datums differ from one site to another, but several datum levels are within 1 m.y. at all sites. First and last appearances are most numerous in two intervals, the late Eocene to early Oligocene (about 32 to 37 Ma) and the early to middle Miocene (about 13 to 18.5 Ma). Isotopic events occur within each of these periods of benthic faunal change, but the isotopic events have a shorter duration and start after the initiation of the changes in the fauna. Changes in deep-sea benthic faunal composition are not directly related to short-term oceanographic changes as expressed in isotopic records.

Sediment fraction analysis and population density of living larger foraminifera in the Persian Gulf (Table 1A, 1B)

Living Heterostegina depressa were found in the Persian Gulf on shallows and sides of islands in the Central Basin. Preliminary culture experiments furnished information on life span, salinity tolerances and population density of species. Reproduction processes (probably asexual) could be observed several times. A possible carbonate production of ca. 150 g/year/m**2 has been estimated.

Abundance of living coccolithophores in the northern Arabian Sea (Table 2)

The present study was conducted to provide information about living coccolithophores from the northern Arabian Sea as potential proxies in palaeoceanographic studies. In all, 71 plankton samples from 16 stations collected in September 1993 were analysed for their contents of living coccolithophores. Absolute abundances range from less than 400 coccospheres per litre in surface waters to 35 000 spheres per litre at intermediate water depths. From 49 identified taxa, nine species contribute significant cell numbers of more than 2000 coccospheres per litre and comprise more than 10% of the communities in at least one sample. Important species are (in approximate order of cell abundances): Gephyrocapsa oceanica, Florisphaera profunda, Oolithotus antillarum, Calciosolenia murrayi, Umbellosphaera irregularis, Emiliania huxleyi, Umbellosphaera tenuis, Calciopappus rigidus, and Algirosphaera robusta. At most profiles, a vertical succession of coccolithophore species was found. Calciosolenia murrayi and C. rigidus were restricted to surface waters, whereas high numbers of F. profunda and A. robusta occurred at depths below 40 m. The coccolithophore communities reflected the local oceanographic situation and seemed to be more dependent on mixed layer depth and nutrient availability than on temperature and salinity changes. Additionally, synecologic competition with diatoms in part controlled the species composition and generally reduced the abundance of coccolithophores. Synecological and ecological tolerances of species were discussed with the help of cluster analysis.

Population and life-stage specific sensitivities to temperature and salinity stress in barnacles

Temperature and salinity shape the distribution and genetic structure of marine communities. Future warming and freshening will exert an additional stress to coastal marine systems. The extent to which organisms respond to these shifts will, however, be mediated by the tolerances of all life-stages and populations of species and their potential to adapt. We investigated nauplius and cypris larvae of the barnacle Balanus (Amphibalanus) improvisus from the Swedish west coast with respect to temperature (12, 20, and 28 °C) and salinity (5, 15, and 30) tolerances. Warming accelerated larval development and increased overall survival and subsequent settlement success. Nauplii developed and metamorphosed best at intermediate salinity. This was also observed in cypris larvae when the preceding nauplii stages had been reared at a salinity of 30. Direct comparisons of the present findings with those on a population from the more brackish Baltic Sea demonstrate contrasting patterns. We conclude that i) B. improvisus larvae within the Baltic region will be favoured by near-future seawater warming and freshening, that ii) salinity tolerances of larvae from the two different populations reflect salinities in their native habitats, but are nonetheless suboptimal and that iii) this species is generally highly plastic with regard to salinity.

Sedimentology, geochemistry and ostracodes from Lake Ohrid

Ancient Lake Ohrid, located in the southern Balkan Peninsula in Macedonia and Albania is characterized by a high degree of endemism and it is considered to be the oldest lake in Europe. But its exact age (between one and ten million years) and also its origin are so far not known. To unravel these uncertainties an ICDP (International Continental Scientific Drilling Program) drilling project (Scientific Collaboration On Past Speciation Conditions in Ohrid (SCOPSCO)), started in April 2013. In addition to the investigations about the age and origin, other paleolimnological studies, e.g., the reconstruction of past climate and of past lake level changes, should be performed with the drilled cores. Used proxies in such paleolimnological studies are, e.g., ostracodes because they respond sensitively to environmental changes but an accurate knowledge of their preferences and tolerances to specific environmental conditions is necessary for this purpose. So far, this knowledge about the, mostly endemic, Ohrid ostracodes was limited. Thus, within the framework of this thesis, ostracodes and a multiplicity of environmental data were collected in Lake Ohrid and its adjacent waters during four field campaigns. In a total of 47 ostracode species could be detected in the entire study area and 32 of them were found alive in Lake Ohrid. Multivariate statistic identified that water depth, salinity, conductivity, pH, and dissolved oxygen were the main determining factors for ostracode distribution in the entire study area. In Lake Ohrid, the distribution was mainly controlled by water depth, water temperature, and pH. Some ostracodes were identified as strong indicator species for important environmental variables, e.g., water temperature and water depth. A distinctive feature of Lake Ohrid was the finding of the ostracode genus Amnicythere whose species normally inhabit oligo-(meso-)haline waters and this could point to a marine origin of the lake. So far, the specialized endemic ostracodes show the highest abundances and the greatest spatial distribution in Lake Ohrid but during the sampling eight widespread species were found for the first time in the lake. They inhabited mainly the northern part of the lake, where two cities are located and industry and agriculture play a major role, and they were limited to water depths above 50 m and this could be an evidence for an increasing anthropogenic pressure because widespread ostracode species often replace endemic species. To unravel the human impact on Lake Ohrid during the last decades short sediment cores were taken and the multi-proxy study indicated that the lake productivity between the early 1920s and the late 1980s was relatively low. Diatom assemblages indicate a rising productivity in the southern part of Lake Ohrid since the mid 1970s and geochemical proxies and ostracodes point to an increasing productivity since the late 1980s in the southern and in the northern part. A slight increase in the productivity continued until 2009. Noticeable is the fact that since the early 1990s, the increasing productivity and the increasing concentrations of heavy metals correspond to a decreasing number of ostracodes in the northern part of Lake Ohrid. Perhaps, this indicates that living conditions in this lake part became less favorable for the mostly endemic ostracode species. Furthermore, the sediment samples from the cores show relatively high concentrations of arsenic, iron, and nickel. Fluctuations in ostracode assemblages from three longer sediment cores, the longest spans approximately 136 ka, taken in Lake Ohrid, correspond to fluctuations in the productivity, in the carbonate content, of the lake level, and of climate changes. Between the marine isotope stage (MIS) 6 and MIS 2 the number of ostracode valves is very low or the valves were completely absent. This corresponds to a low lake productivity, a low carbonate content, and a low lake level. At the onset of the Holocene, the number of valves increased markedly and this correlates with an increased productivity and carbonate content and a warmer climate. But during the Little Ice Age (LIA), the number of valves dropped again and species which prefer warmer waters disappeared completely. This drop corresponds also to a low productivity. After the LIA, the number of species increased again but since 1895 AD a strong and abrupt decrease is visible. A reason for this could be an increase in the heavy metal concentrations.