Phenotypic plasticity in the monoclonal marbled crayfish is associated with very low genetic diversity but pronounced epigenetic diversity.

Clonal organisms are particularly useful to investigate the contribution of epigenetics to phenotypic plasticity, because confounding effects of genetic variation are negligible. In the last decade, the apomictic parthenogenetic marbled crayfish, Procambarus virginalis, has been developed as a model to investigate the relationships between phenotypic plasticity and genetic and epigenetic diversity in detail. This crayfish originated about 30 years ago by autotriploidy from a single slough crayfish Procambarus fallax. As the result of human releases and active spreading, marbled crayfish has established numerous populations in very diverse habitats in 22 countries from the tropics to cold temperate regions. Studies in the laboratory and field revealed considerable plasticity in coloration, spination, morphometric parameters, growth, food preference, population structure, trophic position, and niche width. Illumina and PacBio whole-genome sequencing of marbled crayfish from representatives of 19 populations in Europe and Madagascar demonstrated extremely low genetic diversity within and among populations, indicating that the observed phenotypic diversity and ability to live in strikingly different environments are not due to adaptation by selection on genetic variation. In contrast, considerable differences were found between populations in the DNA methylation patterns of hundreds of genes, suggesting that the environmentally induced phenotypic plasticity is mediated by epigenetic mechanisms and corresponding changes in gene expression. Specific DNA methylation fingerprints persisted in local populations over successive years indicating the existence of epigenetic ecotypes, but there is presently no information as to whether these epigenetic signatures are transgenerationally inherited or established anew in each generation and whether the recorded phenotypic plasticity is adaptive or nonadaptive.

Paradigm shifts in animal epigenetics: Research on non-model species leads to new insights into dependencies, functions and inheritance of DNA methylation.

Recent investigations with non-model species and whole-genome approaches have challenged several paradigms in animal epigenetics. They revealed that epigenetic variation in populations is not the mere consequence of genetic variation, but is a semi-independent or independent source of phenotypic variation, depending on mode of reproduction. DNA methylation is not positively correlated with genome size and phylogenetic position as earlier believed, but has evolved differently between and within higher taxa. Epigenetic marks are usually not completely erased in the zygote and germ cells as generalized from mouse, but often persist and can be transgenerationally inherited, making them evolutionarily relevant. Gene body methylation and promoter methylation are similar in vertebrates and invertebrates with well methylated genomes but transposon silencing through methylation is variable. The new data also suggest that animals use epigenetic mechanisms to cope with rapid environmental changes and to adapt to new environments. The main benefiters are asexual populations, invaders, sessile taxa and long-lived species.

Studying phenotypic variation and DNA methylation across development, ecology and evolution in the clonal marbled crayfish: a paradigm for investigating epigenotype-phenotype relationships in macro-invertebrates.

Animals can produce different phenotypes from the same genome during development, environmental adaptation and evolution, which is mediated by epigenetic mechanisms including DNA methylation. The obligatory parthenogenetic marbled crayfish, Procambarus virginalis, whose genome and methylome are fully established, proved very suitable to study this issue in detail. Comparison between developmental stages and DNA methylation revealed low expression of Dnmt methylation and Tet demethylation enzymes from the spawned oocyte to the 256 cell embryo and considerably increased expression thereafter. The global 5-methylcytosine level was 2.78% at mid-embryonic development and decreased slightly to 2.41% in 2-year-old adults. Genetically identical clutch-mates raised in the same uniform laboratory setting showed broad variation in morphological, behavioural and life history traits and differences in DNA methylation. The invasion of diverse habitats in tropical to cold-temperate biomes in the last 20 years by the marbled crayfish was associated with the expression of significantly different phenotypic traits and DNA methylation patterns, despite extremely low genetic variation on the whole genome scale, suggesting the establishment of epigenetic ecotypes. The evolution of marbled crayfish from its parent species Procambarus fallax by autotriploidy a few decades ago was accompanied by a significant increase in body size, fertility and life span, a 20% reduction of global DNA methylation and alteration of methylation in hundreds of genes, suggesting that epigenetic mechanisms were involved in speciation and fitness enhancement. The combined analysis of phenotypic traits and DNA methylation across multiple biological contexts in the laboratory and field in marbled crayfish may serve as a blueprint for uncovering the role of epigenetic mechanisms in shaping of phenotypes in macro-invertebrates.

Cytology, function and dynamics of stem and progenitor cells in decapod crustaceans.

Stem cells play key roles in development, tissue homeostasis, regeneration, ageing and diseases. Comprehensive reviews on stem cells are available for the determinately growing mammals and insects and some lower invertebrates like hydra but are rare for larger, indeterminately growing invertebrates that can live for many decades. This paper reviews the cytology, function and dynamics of stem and progenitor cells in the decapod crustaceans, a species-rich and ecologically and economically important animal group that includes mainly indeterminate growers but also some determinate growers. Further advantages of decapods for stem cell research are almost 1000-fold differences in body size and longevity, the regeneration of damaged appendages and the virtual absence of age-related diseases and tumours in the indeterminately growing species. The available data demonstrate that the Decapoda possess a remarkable variety of structurally and functionally different stem cells in embryos and larvae, and in the epidermis, musculature, haematopoietic tissue, heart, brain, hepatopancreas, olfactory sense organs and gonads of adults. Some of these seem to be rather continuously active over a lifetime but others are cyclically activated and silenced in periods of days, weeks and years, depending on the specific organ and function. Stem cell proliferation is triggered by signals related to development, moulting, feeding, reproduction, injury, infection, environmental enrichment and social status. Some regulatory pathways have already been identified, including the evolutionarily conserved GATA-binding and runt-domain transcription factors, the widespread neurotransmitter serotonin, the arthropod-specific hormone 20-hydroxyecdysone and the novel astakine growth factors. Knowledge of stem cells in decapods primarily refines our picture on the development, growth and maintenance of tissues and organs in this animal group. Cultured decapod stem cells have good potential for toxicity testing and virus research with practical relevance for aquaculture. Knowledge of stem cells in decapods also broadens our understanding of the evolution of stem cells and regeneration in the animal kingdom. The stem cells of long-lived, indeterminately growing decapods may hold the key to understanding how stem and progenitor cells function into old age without adverse side effects, possibly evoking new ideas for the development of anti-ageing and anti-cancer treatments in humans.

Environmental Adaptation of Genetically Uniform Organisms with the Help of Epigenetic Mechanisms-An Insightful Perspective on Ecoepigenetics.

Organisms adapt to different environments by selection of the most suitable phenotypes from the standing genetic variation or by phenotypic plasticity, the ability of single genotypes to produce different phenotypes in different environments. Because of near genetic identity, asexually reproducing populations are particularly suitable for the investigation of the potential and molecular underpinning of the latter alternative in depth. Recent analyses on the whole-genome scale of differently adapted clonal animals and plants demonstrated that epigenetic mechanisms such as DNA methylation, histone modifications and non-coding RNAs are among the molecular pathways supporting phenotypic plasticity and that epigenetic variation is used to stably adapt to different environments. Case studies revealed habitat-specific epigenetic fingerprints that were maintained over subsequent years pointing at the existence of epigenetic ecotypes. Environmentally induced epimutations and corresponding gene expression changes provide an ideal means for fast and directional adaptation to changing or new conditions, because they can synchronously alter phenotypes in many population members. Because microorganisms inclusive of human pathogens also exploit epigenetically mediated phenotypic variation for environmental adaptation, this phenomenon is considered a universal biological principle. The production of different phenotypes from the same DNA sequence in response to environmental cues by epigenetic mechanisms also provides a mechanistic explanation for the "general-purpose genotype hypothesis" and the "genetic paradox of invasions".

Synthesis of digestive enzymes, food processing, and nutrient absorption in decapod crustaceans: a comparison to the mammalian model of digestion.

The ∼15.000 decapod crustaceans that are mostly omnivorous have evolved a structurally and functionally complex digestive system. They have highly effective cuticular chewing and filtering structures in the stomach, which are regularly renewed by moulting. Decapods produce a broad range of digestive enzymes including chitinases, cellulases, and collagenases with unique properties. These enzymes are synthesized in the F-cells of the hepatopancreas and are encoded in the genome as pre-pro-proteins. In contrast to mammals, they are stored in a mature form in the lumen of the stomach to await the next meal, and therefore, the enzymes are particularly stable. The fat emulsifiers are fatty acyl-dipeptides rather than bile salts. After mechanical and chemical processing of the food in the cardiac stomach, the chyme is filtered by two unique filter systems of different mesh-size. The filtrate is then transferred to the hepatopancreas where the nutrients are absorbed by the R-cells, mostly via carriers, resembling nutrient absorption in the small intestine of mammals. The absorbed nutrients are used to fuel the metabolism of the hepatopancreas, are supplied to other organs, and are stored in the R-cells as glycogen and lipid reserves. Export lipids are secreted from the R-cells into the haemolymph as high density lipoproteins that mainly consist of phospholipids. In contrast to mammals, the midgut tube and hindgut contribute only little to food processing and nutrient absorption. The oesophagus, stomach and hindgut are well innervated but the hepatopancreas lacks nerves. Hormone cells are abundant in the midgut and hepatopancreas epithelia. Microorganisms are often present in the intestine of decapods, but they are apparently not essential for digestion and nutrition.

Epigenetic variation in animal populations: Sources, extent, phenotypic implications, and ecological and evolutionary relevance.

Laboratory experiments and fieldwork with asexually reproducing invertebrates and vertebrates clearly revealed that animal populations can produce substantial phenotypic variation despite genetic identity. This epigenetically caused phenotypic variation comes from two different sources, namely directional environmental induction and bed-hedging developmental stochasticity. Both occur together and are mediated by molecular epigenetic mechanisms like DNA methylation, histone modifications and microRNAs. These epigenetic mechanisms are also involved in insect polyphenism, phenotypic changes in early domestication, and gene expression change and chromatin rearrangement during speciation. Epigenetic variation is particularly important for asexual populations helping them to stay in the game of life when the environmental conditions change. However, it is also relevant for sexually reproducing populations, as shown for genetically impoverished invasive groups, cave animals and sessile taxa that cannot evade unfavourable environmental conditions. Experiments revealed that epigenetic marks can be transgenerationally inherited and persist for several generations. First evidence suggests that inherited epimutations with phenotypic effects may end-up in phenotype-fixing genetic mutations by accelerated mutation of methylated nucleotides. Refined concepts, suitable animal models, fast and affordable new omics techniques that require only small tissue samples, and appropriate data interpretation tools are now available enabling future investigations in ecological and evolutionary epigenetics with high accuracy.

Cytopathology and immune response in the hepatopancreas of decapod crustaceans.

The hepatopancreas of decapod crustaceans is used as an example to illustrate the range of cytopathologies, detoxification mechanisms, and immune responses that environmental toxicants and pathogens can induce in a single organ. The hepatopancreas is the central metabolic organ of decapods and consists of hundreds of blindly-ending tubules and intertubular spaces. The tubular epithelium contains 5 structurally and functionally different cell types, and the interstitium contains haemolymph, haemocytes, connective tissue, and fixed phagocytes. Some physiological conditions such as moulting and starvation cause marked but reversible ultrastructural alterations of the epithelial cells. Environmental toxicants induce either detoxification mechanisms or structural damage in cells, depending on toxicant and concentration. The hepatopancreas is also a main target organ for pathogens, mainly viruses, bacteria, and protists that enter the body via the digestive tract and gills and replicate in the hepatopancreatocytes. The cytopathologies caused by toxicants and pathogens affect single cell types specifically or, more often, several cell types simultaneously. Pathogenesis often begins in a certain cell organelle such as the nucleus, mitochondrion, or endoplasmic reticulum, spreads to other organelles, and ends with death of the infected cell. Fixed phagocytes in the interstitium capture and degrade pathogens that move from the infected tubules into the intertubular spaces or enter the hepatopancreas via circulation. Relatively few disease agents elicit the melanisation and encapsulation reaction that encloses infected tubules by a rigid melanised capsule and kills the entrapped pathogens.

How should we treat autopolyploid and parthenogenetic animals taxonomically?

Asexually reproducing and polyploid animals are unresolved problems of taxonomy and nomenclature. The present paper discusses this issue from a theoretical perspective and outlines how the problem was treated in the past. The autotriploid, obligately parthenogenetic marbled crayfish is used as an example to elaborate the conditions under which autopolyploids and parthenogens should be described as separate uniparental species.

Functional cytology of the hepatopancreas of decapod crustaceans.

This article reviews the morphogenesis, morphology, histology, ultrastructure, and structural-functional relationships of the hepatopancreas, the main metabolic organ of the Decapoda. The hepatopancreas develops in early larval stages from a pair of lateral lobes of the midgut anlage. In adults, it consists of hundreds of blindly ending tubules that are enveloped by a muscle net consisting of longitudinal and circular fibers. Stem cells at the distal ends of the tubules give rise to three ultrastructurally different epithelial cell types, the R-, F-, and B-cells. Histochemistry, immunohistochemistry, in situ hybridization, and monitoring of ultrastructural changes under different experimental conditions allowed the attribution of functions to these cell types. R-cells serve for the absorption and metabolization of nutrients, storage of energy reserves and minerals, synthesis of lipoproteins for export to other organs, detoxification of heavy metals, and excretion of uric acid. F-cells synthesize digestive enzymes and blood proteins involved in oxygen transport and immune defense. They also detoxify some heavy metals and probably organic xenobiotics. B-cells are assumed to produce and recycle fat emulsifiers. The hepatopancreas tubules lack nerves. The presence of scattered M-cells with putative endocrine function in the epithelium suggests that the hepatopancreas is mainly hormonally controlled. M-cells probably represent a self-perpetuating cell lineage independent from E-cells. The interstitium between the tubules contains connective tissue, arterioles, hemolymph with circulating hemocytes, and fixed phagocytes that eliminate pathogens. The hepatopancreas is histologically and ultrastructurally uniform throughout the Decapoda, despite their broad variety in body size, morphology, life style, and ecology. However, in a few cavernicolous and deep-sea shrimps parts of the hepatopancreas are transformed into large oil storing and bioluminescent compartments. Within the malacostracan crustaceans, the hepatopancreas of the Decapoda is most similar to the digestive gland of the Euphausiacea, supporting close taxonomic relationship of these two taxa.

Structure, function and development of the digestive system in malacostracan crustaceans and adaptation to different lifestyles.

The digestive system of the malacostracan crustaceans, namely the decapods, isopods, amphipods and mysids, is among the most complex organ systems of the animal kingdom serving multiple functions such as food processing, absorption and storage of nutrients, synthesis of digestive enzymes and blood proteins, detoxification of xenobiotics and osmoregulation. It is rather well investigated compared to other invertebrates because the Malacostraca include many ecological keystone species and food items for humans. The Decapoda and Peracarida share food processing with chewing and filtering structures of the stomach but differ with respect to morphology and ultrastructure of the digestive glands. In the Peracarida, the digestive glands are composed of few, relatively large lateral caeca, whereas in the Decapoda, hundreds to thousands of blindly ending tubules form a voluminous hepatopancreas. Morphogenesis and onset of functionality of the digestive system strongly depend on the mode of development. The digestive system is early developed in species with feeding planktonic larvae and appears late in species with direct lecithotrophic development. Some structures of the digestive system like the stomach ossicles are rather constant in higher taxa and are of taxonomic value, whereas others like the chewing structures are to some degree adapted to the feeding strategy. The nutrient absorbing and storing cells of the digestive glands show considerable ultrastructural variation during moult cycle, vitellogenesis and starvation. Some of the various functions of the digestive system are already assigned to specific sections of the digestive tract and cell types, but others still await precise localization.

Morphological characterization and genotyping of the marbled crayfish and new evidence on its origin.

The obligately parthenogenetic marbled crayfish, Procambarus virginalis, is the first formally described asexual species of the Crustacea Decapoda. It is a triploid descendant of the sexually reproducing slough crayfish, Procambarus fallax. Here we describe the morphology of cultured and wild marbled crayfish of wide size ranges in detail and photodocument all taxonomically relevant characters. Some morphological traits and coloration showed considerable variation within populations despite the monoclonal nature of marbled crayfish. There were also significant differences between wild and laboratory populations with respect to body proportions, coloration and spination. Comparison with Procambarus fallax revealed no qualitative morphological characters that unambiguously identify the marbled crayfish. Analysis of the mitochondrial cytochrome c oxidase subunit I gene (COI) and nuclear microsatellites of marbled crayfish and Procambarus fallax from different sources indicated that the tri-allelic microsatellite PclG-02 is better suitable than COI to identify the marbled crayfish. A respective identification key is provided. The COI and microsatellites of Procambarus fallax from different areas of Florida and southern Georgia suggest that the parents of the first marbled crayfish may have come from northern Union County, northern Florida.

Annotated bibliography of the parthenogenetic marbled crayfish Procambarus virginalis, a new research model, potent invader and popular pet.

The marbled crayfish Procambarus virginalis is a new obligately parthenogenetic species that was detected in the mid-1990s in the German aquarium trade. Since then it has become a popular pet in many countries throughout the world and a valuable laboratory model for a broad range of biological disciplines. Releases have led to the establishment of wild populations in several European countries, Madagascar and probably Japan, making marbled crayfish an interesting paradigm of evolutionarily young and ongoing bioinvasions. This article provides an annotated bibliography of the scientific and popular scientific literature on marbled crayfish from its detection until today. Each reference is assigned to a publication format and one or more biological categories. The content is shortly described and its significance for marbled crayfish research and general biology is assessed. Of the 239 references listed 140 (58.6%) deal primarily with laboratory experiments on the biology of marbled crayfish and the establishment and use of marbled crayfish as a research model, 74 (31.0%) with its biogeography, invasions and ecology and 25 (10.4%) with hobby aquarist issues and the pet trade.

Investigating the genetic and epigenetic basis of big biological questions with the parthenogenetic marbled crayfish: A review and perspectives.

In the last 15 years, considerable attempts have been undertaken to develop the obligately parthenogenetic marbled crayfish Procambarus virginalis as a new model in biology. Its main advantage is the production of large numbers of offspring that are genetically identical to the mother, making this crustacean particularly suitable for research in epigenetics. Now, a draft genome, transcriptome and genome-wide methylome are available opening new windows for research. In this article, I summarize the biological advantages and genomic and epigenetic features of marbled crayfish and, based on first promising data, discuss what this new model could contribute to answering of ''big'' biological questions. Genome mining is expected to reveal new insights into the genetic specificities of decapod crustaceans, the genetic basis of arthropod reproduction, moulting and immunity, and more general topics such as the genetic underpinning of adaptation to fresh water, omnivory, biomineralization, sexual system change, behavioural variation, clonal genome evolution, and resistance to cancer. Epigenetic investigations with the marbled crayfish can help clarifying the role of epigenetic mechanisms in gene regulation, tissue specification, adult stem cell regulation, cell ageing, organ regeneration and disease susceptibility. Marbled crayfish is further suitable to elucidate the relationship between genetic and epigenetic variation, the transgenerational inheritance of epigenetic signatures and the contribution of epigenetic phenotype variation to the establishment of social hierarchies, environmental adaptation and speciation. These issues can be tackled by experiments with highly standardized laboratory lineages, comparison of differently adapted wild populations and the generation of genetically and epigenetically edited strains.

Facilitation of environmental adaptation and evolution by epigenetic phenotype variation: insights from clonal, invasive, polyploid, and domesticated animals.

There is increasing evidence, particularly from plants, that epigenetic mechanisms can contribute to environmental adaptation and evolution. The present article provides an overview on this topic for animals and highlights the special suitability of clonal, invasive, hybrid, polyploid, and domesticated species for environmental and evolutionary epigenetics. Laboratory and field studies with asexually reproducing animals have shown that epigenetically diverse phenotypes can be produced from the same genome either by developmental stochasticity or environmental induction. The analysis of invasions revealed that epigenetic phenotype variation may help to overcome genetic barriers typically associated with invasions such as bottlenecks and inbreeding. Research with hybrids and polyploids established that epigenetic mechanisms are involved in consolidation of speciation by contributing to reproductive isolation and restructuring of the genome in the neo-species. Epigenetic mechanisms may even have the potential to trigger speciation but evidence is still meager. The comparison of domesticated animals and their wild ancestors demonstrated heritability and selectability of phenotype modulating DNA methylation patterns. Hypotheses, model predictions, and empirical results are presented to explain how epigenetic phenotype variation could facilitate adaptation and speciation. Clonal laboratory lineages, monoclonal invaders, and adaptive radiations of different evolutionary age seem particularly suitable to empirically test the proposed ideas. A respective research agenda is presented.

Structural specialties, curiosities, and record-breaking features of crustacean reproduction.

Crustaceans are a morphologically, physiologically, and ecologically highly diverse animal group and correspondingly diverse are their reproductive characteristics. They have evolved structural specialties with respect to penis construction, sperm form, sperm storage, fertilization, and brood care. Unique in the animal kingdom are safety lines that safeguard hatching and first molting. Further curiosities are dwarf males in parasitic and sessile crustaceans and bacteria-induced feminization and gigantism of crustacean hosts. Record-breaking features are relative penis length, sperm size, clutch size, chromosome number, viability of dormant eggs, and fossil ages of penis, sperm, and brooded embryos. These examples from a single invertebrate subphylum and a single life history aspect illustrate that morphological solutions to functional requirements can be as spectacular as behavioral adaptations. They may provide valuable sources for comparative morphologists, ecologists, evolutionary biologists, and applied biologists to advance topical issues such as sperm competition, posthumous paternity, evolution of brood care, adaptation to freshwater, infectious feminization, sustainable male-based fishery, maintenance of genetic diversity under conditions of limited mating opportunity, and long-term impact of pollution on genotype and phenotype. J. Morphol. 277:1399-1422, 2016. © 2016 Wiley Periodicals, Inc.

The marbled crayfish as a paradigm for saltational speciation by autopolyploidy and parthenogenesis in animals.

The parthenogenetic all-female marbled crayfish is a novel research model and potent invader of freshwater ecosystems. It is a triploid descendant of the sexually reproducing slough crayfish, Procambarus fallax, but its taxonomic status has remained unsettled. By cross-breeding experiments and parentage analysis we show here that marbled crayfish and P. fallax are reproductively separated. Both crayfish copulate readily, suggesting that the reproductive barrier is set at the cytogenetic rather than the behavioural level. Analysis of complete mitochondrial genomes of marbled crayfish from laboratory lineages and wild populations demonstrates genetic identity and indicates a single origin. Flow cytometric comparison of DNA contents of haemocytes and analysis of nuclear microsatellite loci confirm triploidy and suggest autopolyploidisation as its cause. Global DNA methylation is significantly reduced in marbled crayfish implying the involvement of molecular epigenetic mechanisms in its origination. Morphologically, both crayfish are very similar but growth and fecundity are considerably larger in marbled crayfish, making it a different animal with superior fitness. These data and the high probability of a divergent future evolution of the marbled crayfish and P. fallax clusters suggest that marbled crayfish should be considered as an independent asexual species. Our findings also establish the P. fallax-marbled crayfish pair as a novel paradigm for rare chromosomal speciation by autopolyploidy and parthenogenesis in animals and for saltational evolution in general.

Stochastic developmental variation, an epigenetic source of phenotypic diversity with far-reaching biological consequences.

This article reviews the production of different phenotypes from the same genotype in the same environment by stochastic cellular events, nonlinear mechanisms during patterning and morphogenesis, and probabilistic self-reinforcing circuitries in the adult life. These aspects of phenotypic variation are summarized under the term 'stochastic developmental variation' (SDV) in the following. In the past, SDV has been viewed primarily as a nuisance, impairing laboratory experiments, pharmaceutical testing, and true-to-type breeding. This article also emphasizes the positive biological effects of SDV and discusses implications for genotype-to-phenotype mapping, biological individuation, ecology, evolution, and applied biology. There is strong evidence from experiments with genetically identical organisms performed in narrowly standardized laboratory set-ups that SDV is a source of phenotypic variation in its own right aside from genetic variation and environmental variation. It is obviously mediated by molecular and higher-order epigenetic mechanisms. Comparison of SDV in animals, plants, fungi, protists, bacteria, archaeans, and viruses suggests that it is a ubiquitous and phylogenetically old phenomenon. In animals, it is usually smallest for morphometric traits and highest for life history traits and behaviour. SDV is thought to contribute to phenotypic diversity in all populations but is particularly relevant for asexually reproducing and genetically impoverished populations, where it generates individuality despite genetic uniformity. In each generation, SDV produces a range of phenotypes around a well-adapted target phenotype, which is interpreted as a bet-hedging strategy to cope with the unpredictability of dynamic environments. At least some manifestations of SDV are heritable, adaptable, selectable, and evolvable, and therefore, SDV may be seen as a hitherto overlooked evolution factor. SDV is also relevant for husbandry, agriculture, and medicine because most pathogens are asexuals that exploit this third source of phenotypic variation to modify infectivity and resistance to antibiotics. Since SDV affects all types of organisms and almost all aspects of life, it urgently requires more intense research and a better integration into biological thinking.

Abbreviation of larval development and extension of brood care as key features of the evolution of freshwater Decapoda.

The transition from marine to freshwater habitats is one of the major steps in the evolution of life. In the decapod crustaceans, four groups have colonized fresh water at different geological times since the Triassic, the freshwater shrimps, freshwater crayfish, freshwater crabs and freshwater anomurans. Some families have even colonized terrestrial habitats via the freshwater route or directly via the sea shore. Since none of these taxa has ever reinvaded its environment of origin the Decapoda appear particularly suitable to investigate life-history adaptations to fresh water. Evolutionary comparison of marine, freshwater and terrestrial decapods suggests that the reduction of egg number, abbreviation of larval development, extension of brood care and lecithotrophy of the first posthatching life stages are key adaptations to fresh water. Marine decapods usually have high numbers of small eggs and develop through a prolonged planktonic larval cycle, whereas the production of small numbers of large eggs, direct development and extended brood care until the juvenile stage is the rule in freshwater crayfish, primary freshwater crabs and aeglid anomurans. The amphidromous freshwater shrimp and freshwater crab species and all terrestrial decapods that invaded land via the sea shore have retained ocean-type planktonic development. Abbreviation of larval development and extension of brood care are interpreted as adaptations to the particularly strong variations of hydrodynamic parameters, physico-chemical factors and phytoplankton availability in freshwater habitats. These life-history changes increase fitness of the offspring and are obviously favoured by natural selection, explaining their multiple origins in fresh water. There is no evidence for their early evolution in the marine ancestors of the extant freshwater groups and a preadaptive role for the conquest of fresh water. The costs of the shift from relative r- to K-strategy in freshwater decapods are traded-off against fecundity, future reproduction and growth of females and perhaps against size of species but not against longevity of species. Direct development and extension of brood care is associated with the reduction of dispersal and gene flow among populations, which may explain the high degree of speciation and endemism in directly developing freshwater decapods. Direct development and extended brood care also favour the evolution of social systems, which in freshwater decapods range from simple subsocial organization to eusociality. Hermaphroditism and parthenogenesis, which have evolved in some terrestrial crayfish burrowers and invasive open water crayfish, respectively, may enable populations to adapt to restrictive or new environments by spatio-temporal alteration of their socio-ecological characteristics. Under conditions of rapid habitat loss, environmental pollution and global warming, the reduced dispersal ability of direct developers may turn into a severe disadvantage, posing a higher threat of extinction to freshwater crayfish, primary freshwater crabs, aeglids and landlocked freshwater shrimps as compared to amphidromous freshwater shrimps and secondary freshwater crabs.

Hidden treasures in stem cells of indeterminately growing bilaterian invertebrates.

Indeterminate growth, the life-long growth without fixed limits, is typical of some evolutionarily very successful aquatic invertebrate groups such as the decapod crustaceans, bivalve molluscs and echinoderms. These animals enlarge their organs also in the adult life period and can regenerate lost appendages and organs, which is in sharp contrast to mammals and most insects. Interestingly, decapods, bivalves and echinoderms develop only rarely neoplastic and age-related diseases, although some species reach ages exceeding 100 years. Their stem cell systems must have co-evolved with these successful life histories suggesting possession of unknown and beneficial features that might open up new vistas in stem cell biology. Research of the last decade has identified several adult stem cell systems in these groups and also some mature cell types that are capable to dedifferentiate into multipotent progenitor cells. Investigation of stem and progenitor cells in indeterminately growing bilaterian invertebrates is assumed beneficial for basic stem cell biology, aquaculture, biotechnology and perhaps medicine. The biggest treasure that could be recovered in these animal taxa concerns maintenance of stem cell niches and fidelity of stem cell division for decades without undesirable side effects such as tumour formation. Uncovering of the underlying molecular and regulatory mechanisms might evoke new ideas for the development of anti-ageing and anti-cancer interventions in humans.