Tom Pinceel

Toward a Global Phylogeny of the “Living Fossil" Crustacean Order of the Notostraca

Tadpole shrimp (Crustacea, Notostraca) are iconic inhabitants of temporary aquatic habitats worldwide. Often cited as prime examples of evolutionary stasis, surviving representatives closely resemble fossils older than 200 mya, suggestive of an ancient origin. Despite significant interest in the group as ‘living fossils’ the taxonomy of surviving taxa is still under debate and both the phylogenetic relationships among different lineages and the timing of diversification remain unclear. We constructed a molecular phylogeny of the Notostraca using model based phylogenetic methods. Our analyses supported the monophyly of the two genera Triops and Lepidurus, although for Triops support was weak. Results also revealed high levels of cryptic diversity as well as a peculiar biogeographic link between Australia and North America presumably mediated by historic long distance dispersal. We concluded that, although some present day tadpole shrimp species closely resemble fossil specimens as old as 250 mya, no molecular support was found for an ancient (pre) Mesozoic radiation. Instead, living tadpole shrimp are most likely the result of a relatively recent radiation in the Cenozoic era and close resemblances between recent and fossil taxa are probably the result of the highly conserved general morphology in this group and of homoplasy.

Long Distance Dispersal of Zooplankton Endemic to Isolated Mountaintops - an Example of an Ecological Process Operating on an Evolutionary Time Scale

Recent findings suggest a convergence of time scales between ecological and evolutionary processes which is usually explained in terms of rapid micro evolution resulting in evolution on ecological time scales. A similar convergence, however, can also emerge when slow ecological processes take place on evolutionary time scales. A good example of such a slow ecological process is the colonization of remote aquatic habitats by passively dispersed zooplankton. Using variation at the protein coding mitochondrial COI gene, we investigated the balance between mutation and migration as drivers of genetic diversity in two Branchipodopsis fairy shrimp species (Crustacea, Anostraca) endemic to remote temporary rock pool clusters at the summit of isolated mountaintops in central South Africa. We showed that both species colonized the region almost simultaneously c. 0.8 My ago, but exhibit contrasting patterns of regional genetic diversity and demographic history. The haplotype network of the common B. cf. wolfi showed clear evidence of 11 long distance dispersal events (up to 140 km) with five haplotypes that are shared among distant inselbergs, as well as some more spatially isolated derivates. Similar patterns were not observed for B. drakensbergensis presumably since this rarer species experienced a genetic bottleneck. We conclude that the observed genetic patterns reflect rare historic colonization events rather than frequent ongoing gene flow. Moreover, the high regional haplotype diversity combined with a high degree of haplotype endemicity indicates that evolutionary- (mutation) and ecological (migration) processes in this system operate on similar time scales.

Mechanistic and evolutionary aspects of light-induced dormancy termination in a temporary pond crustacean

Abstract. Most freshwater zooplankton species produce dormant eggs or other long-lived resistant life stages to bridge periods that are unsuitable for growth and reproduction. One of the principal cues that determine activation of these dormant stages is light exposure. Contrary to the analogous process of seed germination in plants, relatively little is known about the mechanism of light activation and its potential adaptive value in freshwater zooplankton. We made use of a temporary pool model to investigate the hatching response of resting eggs of the fairy shrimp Branchipodopsis wolfi to a specific set of selected light regimes and to evaluate the importance of egg pigmentation for mediating susceptibility to light cues. Our results suggest the presence of a critical light threshold for activation leading to maximal hatching. Moreover, the process of light activation does not appear to be purely cumulative because light administered in a single continuous pulse resulted in much higher hatching than the same amount of light administered in several 2-h pulses. Last, darkly pigmented eggs were less sensitive than lightly pigmented eggs to light exposure, resulting in delayed hatching of darker eggs during an experimental inundation. This result indicates that natural variation in egg pigmentation could be a risk-spreading mechanism that trades off the risks of mortality caused by kin competition during the nutrient-limited conditions that typically prevail early during inundations on one hand and the risk of mortality from premature pond drying on the other hand. Overall, our results indicate that light-energy activation thresholds and variation in pigmentation of the eggshell can be important traits that may contribute to variation in hatching phenology among zooplankton lineages.

Fairy shrimps in distress: a molecular taxonomic review of the diverse fairy shrimp genus Branchinella (Anostraca: Thamnocephalidae) in Australia in the light of ongoing environmental change

Australia, and especially South-Western Australia, is a diversity hotspot for large branchiopod crustaceans. A significant proportion of this diversity is found in the anostracans (Crustacea, Anostraca) and particularly in the diverse genus Branchinella with at least 34 species. Members of this genus are found exclusively in temporary aquatic habitats which are increasingly threatened by secondary salinization and other anthropogenic pressures. The development of adequate conservation strategies is therefore considered a priority. To define conservation units, however, thorough knowledge of the taxonomy and phylogenetic position of extant lineages is essential. We reconstructed a large scale phylogeny of the Australian Branchinella by analyzing the 16S mitochondrial gene of 31 presumed species, complemented with analysis of morphological structures holding taxonomic information. Results revealed the presence of at least three new cryptic species. On the other hand, some Branchinella lineages, surviving in environments subjected to contrasting selection regimes, appeared to be conspecific. This suggests substantial physiological plasticity or important adaptive variation present in some species, potentially enabling them to better cope with environmental change, such as secondary salinization. Overall, these results further illustrate the benefits of combining molecular markers and classic morphological taxonomy and phylogeny to assess biodiversity and define conservation units in cryptic groups.

Modelling the sensitivity of life history traits to climate change in a temporary pool crustacean

Temporary pool inhabitants face altered inundation regimes under climate change. While their exposure to these changes has received considerable attention, few studies have investigated their sensitivity or adaptability. Here, we use zooplankton as a model to explore how decreasing hydroperiods affect extinction risks and assess whether changes in life history traits could promote persistence. For this, we construct a three-stage matrix population model parameterised with realistic life-history values for the fairy shrimp Branchipodopsis wolfi from pools with varying hydroperiods. Our results suggest that extinction risks increase drastically once the median hydroperiod drops below a critical threshold. Although changes in life-history parameters could potentially compensate for this risk, the relative importance of each trait for population growth depends on the median hydroperiod. For example, survival of dormant eggs seemed to be most important when hydroperiods were short while the survival of freshly laid eggs and adult individuals were more important in longer-lived pools. Overall, this study demonstrates that zooplankton species are sensitive to climate change and that the adaptive capacity of organisms from temporary pools with dissimilar hydrology hinges on selection of different life history traits.

Dormancy and dispersal as mediators of zooplankton population and community dynamics along a hydrological disturbance gradient in inland temporary pools

At some stage in their life cycle, most zooplankton in temporary waters produce dormant eggs that assemble in a persistent egg bank to cope with unfavourable conditions. As part of a risk-spreading strategy, only a fraction of the egg bank hatches during a single inundation. Besides this dispersal in time, resistant dormant eggs also disperse in space via vectors including wind, water and animals. The structure and functioning of the dormant egg bank has important consequences for (meta) population and (meta) community structure and dynamics. Here, we merge empirical and theoretical data into a conceptual framework for the study of population and community responses in temporary ponds along a gradient in hydrological disturbance. Overall, we conclude that changes in hydrological disturbance may compromise both the abiotic (i.e. water quality) and biotic (i.e. population and community processes) integrity of temporary pools which is especially relevant in light of ongoing anthropogenic alterations in the hydrology of inland waters.

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri

The killifish Nothobranchius furzeri is an emerging model organism in the field of ecotoxicology and its applicability in acute and chronic ecotoxicity testing has been demonstrated. Overall, the sensitivity of the species to toxic compounds is in the range with, or higher than, that of other model species. This work describes protocols for acute, chronic, and multigenerational bioassays of single and combined stressor effects on N. furzeri. Due to its short maturation time and life-cycle, this vertebrate model allows the study of endpoints such as maturation time and fecundity within four months. Transgenerational full life-cycle exposure trials can be performed in as little as 8 months. Since this species produces eggs that are drought-resistant and remain viable for years, the on-site culture of the species is not needed but individuals can be recruited when required. The protocols are designed to measure life-history traits (mortality, growth, fecundity, weight) and critical thermal maximum.

Improving the reliability and ecological validity of pharmaceutical risk assessment – Turquoise killifish (Nothobranchius furzeri) as a model in behavioural ecotoxicology

Pharmaceuticals are essential for human well-being, but their increasing and continuous use pollutes the environment. Although behavioral ecotoxicology is increasingly advocated to assess the effects of pharmaceutical pollution on wildlife and ecosystems, a consensus on the actual environmental risks is lacking for most compounds. The main limitation is the lack of standardized reproducible tests that are based on sensitive behavioral endpoints and that accommodate a high ecological relevance. In the present study, we assessed the impact of a 3-wk exposure to the antidepressant fluoxetine on multiple behavioral traits in the promising new model organism Nothobranchius furzeri (turquoise killifish). Overall, our study shows that fluoxetine can impact feeding behavior, habitat choice in a novel environment, and antipredator response of N. furzeri individuals; effects on spontaneous activity and exploration tendency were less pronounced. However, effects became only apparent when individuals were exposed to fluoxetine concentrations that were 10 times higher than typical concentrations in natural aquatic environments. Ecotoxicologists are challenged to maximize both the reliability and ecological validity of risk assessments of pollutants. Our study contributes to the development of a time- and cost-efficient, standardized ecotoxicological test based on sensitive, ecologically relevant behavioral endpoints in N. furzeri. Environ Toxicol Chem 2019;38:262-270.

Combined effects of cadmium exposure and temperature on the annual killifish (Nothobranchius furzeri)

Freshwater organisms are increasingly exposed to combinations of stressors. However, because it is time-consuming and costly, research on the interaction of stressors, such as compound toxicity and global warming on vertebrates, is scarce. Studies on multigenerational effects of these combined stressors are almost nonexistent. In the present study, we tested the combined effects of 4 °C warming and cadmium (Cd) exposure on life-history traits, biomarkers, bioaccumulation, and multigenerational tolerance in the turquoise killifish, Nothobranchius furzeri. The extremely short life cycle of this vertebrate model allows for assessment of sublethal and multigenerational effects within 4 mo. The applied Cd concentrations had only limited effects on the measured endpoints, which suggests that N. furzeri is more resistant to Cd than fathead minnow and rainbow trout. In contrast, the temperature increase of 4 °C was stressful: it delayed female maturation and lowered adult mass and fecundity. Finally, indications of synergistic effects were found on peak fecundity and embryonic survival. Overall, these results indicate the importance of studying chronic and multigenerational effects of combined stressors. Environ Toxicol Chem 2018;37:2361-2371.