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Background: For over a century, scientists have studied host-pathogen interactions between the crayfish plague disease agent Aphanomyces astaci and freshwater crayfish. It has been hypothesised that North American crayfish hosts are disease-resistant due to the long-lasting coevolution with the pathogen. Similarly, the increasing number of latent infections reported in the historically sensitive European crayfish hosts seems to indicate that similar coevolutionary processes are occurring between European crayfish and A. astaci. Our current understanding of these host-pathogen interactions is largely focused on the innate immunity processes in the crayfish haemolymph and cuticle, but the molecular basis of the observed disease-resistance and susceptibility remain unclear. To understand how coevolution is shaping the host’s molecular response to the pathogen, susceptible native European noble crayfish and invasive disease-resistant marbled crayfish were challenged with two A. astaci strains of different origin: a haplogroup A strain (introduced to Europe at least 50 years ago, low virulence) and a haplogroup B strain (signal crayfish in lake Tahoe, USA, high virulence). Here, we compare the gene expression profiles of the hepatopancreas, an integrated organ of crayfish immunity and metabolism.
Results: We characterised several novel innate immune-related gene groups in both crayfish spe cies. Across allchallenge groups, we detected 412 differentially expressed genes (DEGs) in the noble crayfish, and 257 DEGs in the marbled crayfish. In the noble crayfish, a clear immune response was detected to the haplogroup B strain, but not to the haplogroup A strain. In contrast, in the marbled crayfish we detected an immune response to the haplogroup A strain, but not to the haplogroup B strain. Conclusions: We highlight the hepatopancreas as an important hub for the synthesis of immune molecules in the response to A. astaci. A clear distinction between the innate immune response in the marbled crayfish and the noble crayfish is the capability of the marbled crayfish to mobilise a higher variety of innate immune response effectors.
Objectives: Crayfish plague disease, caused by the oomycete pathogen Aphanomyces astaci represents one of the greatest risks for the biodiversity of the freshwater crayfish. This data article covers the de novo transcriptome assembly and annotation data of the noble crayfish and the marbled crayfish challenged with Ap. astaci. Following the controlled infection experiment (Francesconi et al. in Front Ecol Evol, 2021, https://doi.org/10.3389/fevo.2021.647037), we conducted a differential gene expression analysis described in (Boštjančić et al. in BMC Genom, 2022, https://doi.org/10.1186/s12864-022-08571-z) Data description: In total, 25 noble crayfish and 30 marbled crayfish were selected. Hepatopancreas tissue was isolated, followed by RNA sequencing using the Illumina NovaSeq 6000 platform. Raw data was checked for quality with FastQC, adapter and quality trimming were conducted using Trimmomatic followed by de novo assembly with Trinity. Assembly quality was assessed with BUSCO, at 93.30% and 93.98% completeness for the noble crayfish and the marbled crayfish, respectively. Transcripts were annotated using the Dammit! pipeline and assigned to KEGG pathways. Respective transcriptome and raw datasets may be reused as the reference transcriptome assemblies for future expression studies.
Worldwide one third to one half of the freshwater crayfish species are threatened with population decline or extinction. Besides habitat deterioration, pollution, and other man-made environmental changes, invasive species and pathogens are major threats to the survival of European crayfish species. Freshwater crayfish are the largest freshwater invertebrates and strongly influence the structure of food webs. The disappearance of crayfish from a water body may change the food web and could have dramatic consequences for an ecosystem.rnOne goal in modern species conservation strategies is the conservation of genetic diversity, since genetic diversity is an advantage for the long-term survival of a species. The main aim of my thesis was to reveal the genetic structure and to identify genetic hotspots of the endangered noble crayfish (Astacus astacus) throughout Europe (part 1 of my thesis). Since the most significant threat to biodiversity of European crayfish species is the crayfish plague pathogen Aphanomyces astaci I studied new aspects in the distribution of A. astaci (part two of my thesis). The results serve as a basis for future conservation programs for freshwater crayfish. In the first part of my thesis I conducted a phylogeographic analysis of noble crayfish using mitochondrial DNA and nuclear microsatellite data. With these methods I aimed to identify its genetic hotspots and to reconstruct the recolonization history of central Europe by this species. I detected high genetic diversities in southestern Europe indicating that noble crayfish outlasted the cold climate phases during the Pleistocene in this region (Appendix 1). Because of the high genetic diversity found there, southeastern Europe is of particular importance for the conservation of noble crayfish. The mitochondrial DNA analysis points to a bifurcated colonization process from the eastern Black Sea basin to a) the North Sea and to b) the Baltic Sea basin (Appendix 2). A second independent refugium that was localized on the Western Balkans did not contribute to the colonization of central Europe. Furthermore, I found that the natural genetic structure is dissolved, probably due to the high human impact on the distribution of noble crayfish (e.g. artificial translocation). In the second part of this thesis using real-time PCR I identified calico crayfish (Orconectes immunis) as the fourth North American crayfish species to be carrier of the agent of the crayfish plague (Appendix 3). Furthermore I detected the crayfish plague pathogen in American spiny-cheek crayfish (Orconectes limosus) and native narrow-clawed crayfish (Astacus leptodactylus) in the lower Danube in Romania (Appendix 4). The distribution of infected spiny-cheek crayfish poses a threat to the native biodiversity in southeastern Europe and shows the high invasion potential of this crayfish species. Moreover, I found that even the native narrow-clawed crayfish in the Danube Delta, about 970 km downstream of the current invasion front of American crayfish, is a carrier of A. astaci (Appendix 5). This finding is of high importance, as the native species do not seem to suffer from the infection. In Appendix 6 I elucidate demonstrate that the absence of the crayfish plague agent is the most likely explanation for the coexistence of populations of European and American crayfish in central Europe. In my thesis I show that the common assumption that all North American crayfish are carrier of A. astaci and that all native crayfish species die when infected with A. astaci does not hold true. The studies presented in my thesis reveal new aspects that are crucial for native crayfish conservation: 1) The genetic diversity of noble crayfish is highest in southeastern Europe where noble crayfish outlasted the last glacial maximum in at least two different refugia. 2) Not all American crayfish populations are carrier of A. astaci and 3) not all Europen crayish populations die shortly after being infected with the crayfish plague pathogen.rnTo conserve native crayfish species and their (genetic) diversity in the long term, further introductions of American crayfish into European waters must be avoided. However, the introduction will only decrease if the commercial trade with non-indigenous crayfish species is prohibited.