This study applies molecular and morphological approaches addressing the identification of morphologically similar larval stages (cercariae) of Plagiorchis spp. (Digenea: Plagiorchiidae) parasitising lymnaeid snail populations in the freshwater ecosystems of central Europe. Five morphologically homogeneous and genetically distinct lineages of Plagiorchis spp. were identified via matching molecular data for the mitochondrial cox1 gene with detailed morphometric data. Phylogenetic and comparative sequence analyses using partial 28S rDNA and ITS1-5.8S-ITS2 sequences allowed molecular identification of three species (P. elegans, P. maculosus and P. koreanus) via matching sequences from larval and adult digenean stages. A key for the identification of the cercariae of Plagiorchis spp. parasitising lymnaeid populations in central Europe is provided.

No description available.

Fascioliasis is a worldwide emerging snail-borne zoonotic trematodiasis with a great spreading capacity linked to animal and human movements, climate change, and anthropogenic modifications of freshwater environments. South America is the continent with more human endemic areas caused by Fasciola hepatica, mainly in high altitude areas of Andean regions. The Peruvian Cajamarca area presents the highest human prevalences reported, only lower than those in the Bolivian Altiplano. Sequencing of the complete rDNA ITS-2 allowed for the specific and haplotype classification of lymnaeid snails collected in seasonal field surveys along a transect including 2007–3473 m altitudes. The species Galba truncatula (one haplotype preferentially in higher altitudes) and Pseudosuccinea columella (one haplotype in an isolated population), and the non-transmitting species Lymnaea schirazensis (two haplotypes mainly in lower altitudes) were found. Climatic seasonality proved to influence G. truncatula populations in temporarily dried habitats, whereas L. schirazensis appeared to be more climatologically independent due to its extreme amphibious ecology. Along the southeastern transect from Cajamarca city, G. truncatula and L. schirazensis shared the same site in 7 localities (46.7% of the water collections studied). The detection of G. truncatula in 11 new foci (73.3%), predominantly in northern localities closer to the city, demonstrate that the Cajamarca transmission risk area is markedly wider than previously considered. Lymnaea schirazensis progressively increases its presence when moving away from the city. Results highlight the usefulness of lymnaeid surveys to assess borders of the endemic area and inner distribution of transmission foci. Similar lymnaeid surveys are still in need to be performed in the wide northern and western zones of the Cajamarca city. The coexistence of more than one lymnaeid transmitting species, together with a morphologically indistinguishable non-transmitting species and livestock movements inside the area, conform a complex scenario which poses difficulties for the needed One Health control intervention. / Ministerio de Economía y Competitividad / Revisión por pares

No description available.

Calmodulin (CaM) is a ubiquitous calcium-binding protein responsible for the binding and activation of a vast number of enzymes and signaling pathways. It contains two lobes that bind two calcium ions each, separated by a flexible central linker. This structural flexibility allows CaM to bind and regulate a large number of diverse protein targets within the cell in response to Ca2+ gradients. Voltage gated calcium channels (CaVs), as main sources of extracellular Ca2+, are crucial for a number of physiological processes, from muscle contraction to neurotransmission and endocrine function. These large transmembrane proteins open in response to membrane depolarization and allow gated entry of Ca2+ ions into the cytoplasm. Their regulation is currently the subject of intense investigation due to its pharmacological and scientific importance. CaM has been previously shown to pre-associate and act as a potent inhibitor of one class of high-voltage activated (HVA) channels called L-type channels via its interaction with their C-terminal cytoplasmic region. This interaction is primarily mediated by a conserved CaM-binding motif called the ‘IQ’ motif (for conserved isoleucine and glutamine residues), although the exact molecular details of its involvement in inactivation are currently unclear. Elucidation of these details was the primary objective of this dissertation. Recently, a novel sequence motif within this channel called ‘NSCaTE’ (N-terminal spatial calcium transforming element) has been described as an important contributor to calcium-dependent inactivation (CDI) of L-type channels. It was presumed to be unique to vertebrates, but we also show its conservation in a distantly related L-type channel homolog of Lymnaea stagnalis (pond snail). The interaction of CaM with a number of peptides representing the different regulatory motifs (IQ and NSCaTE) for both mammalian and snail isoforms was characterized in an attempt to better understand their role in CDI. Biophysical work with peptides as well as electrophysiology recordings with an N-terminal truncation mutant of Lymnaea CaV1 homolog were performed to expand our understanding of how the interplay between these channel elements might occur. In brief, the most striking feature of the interaction concerns the strong evidence for a CaM-mediated bridge between the N- and C-terminal elements of L-type channels. Further investigation of the CaM interaction with both IQ and NSCaTE peptides using Ca2+-deficient CaM mutants reveals a preference of both peptides for the Ca2+-C-lobe of CaM, and a much higher affinity of CaM for the IQ peptide, suggesting that the N-lobe of CaM is the main interaction responsible for the physiological effects of NSCaTE. These results are consistent with our electrophysiology findings that reveal a distinct buffer-sensitive CDI in wild type LCaV1 that can be abolished by the N-terminal truncation spanning the NSCaTE region. In addition to L-type channels, CaM has also been shown to have an indirect role in the regulation of low-voltage activated (LVA) or T-type channels (CaV3.x), via their phosphorylation by CaM-dependent protein kinase II (CaMKII). Using a primary sequence scanning algorithm, a CaM-binding site was predicted in a cytoplasmic region of these channels that was also previously shown to be important in channel gating. Biophysical experiments with synthetic peptides spanning this gating brake region from the three human and the single Lymnaea isoform strongly suggest that there is a novel, bona fide CaM interaction in this channel region, and also hint that this interaction may be a Ca2+-dependent switch of some sort. The results confirm a possible new role for CaM in the direct regulation of these channels, although the exact mechanism remains to be elucidated.

Molluscs as well as all other invertebrates rely on innate immune response only. Their internal defense system is capable of destroying most pathogens. However, there are some exceptions, e.g. some snails serve as intermediate hosts for some trematodes. Trematodes are able to develop inside these snails due to intervention in the snail internal defense system. The submitted thesis describes hemocyte activities of two lymnaeid snails, Lymnaea stagnalis a Radix lagotis, and the influence of Trichobilharzia regenti infection on R. lagotis hemocytes. Hemocytes of both species exposed to various chemicals produced different amounts of H2O2 and NO. The response varied between both lymnaeid species. The amount of circulating hemocytes was elevated in R. lagotis snails due to T. regenti infection. However, the infenction attenuated hemocyte activities monitored by us. Hemocyte basal NO production was decreased as well as phagocytosis of bacteria, cell adherence and pseudopodia formation. Toxicity of L. stagnalis plasma against T. regenti miracidia was also described. Mechanisms used by trematodes to interact with the snail internal defense system will help us to understand why one species is suitable for the develepment of the trematode whereas another closely related species kills it. Powered by TCPDF (www.tcpdf.org)

Pour valider une suite de biomarqueurs cellulaires en milieu dulçaquicole, nous avons étudié la structure des systèmes lysosomal et peroxysomal et les variations de contenus en lipofuscines et lipides neutres insaturés dans les tissus digestifs de plusieurs invertébrés, à travers une approche couplant histochimie et analyse d'images. Les expérimentations in situ et bioessais à plus ou moins long terme ont montré la pertinence des réponses cellulaires étudiées pour la mise en évidence de gradients de contamination. Les profils de réponse les plus communs chez les mollusques incluent un gonflement lysosomal, des déplétions de lipides neutres insaturés, une accumulation de lipofuscines et/ou un système peroxysomal plus volumineux. Ces réponses cellulaires précoces peuvent être associées, au niveau individuel, à une désynchronisation de la reproduction entre individus ou à une diminution de l'indice de condition. L'étude mensuelle à long terme a souligné l'absence de niveaux de base saisonniers de ces biomarqueurs et l'influence importante de la disponibilité trophique sur les contenus tissulaires en lipides neutres. Une mise en dépuration de 15 jours du bivalve, Dreissena polymorpha, sur son site d'origine, n'a pas montré de réversibilité du profil des réponses cellulaires à un gradient de contamination in situ, mais elle a tout de même de limité la déplétion des lipides neutres. Les réponses cellulaires les plus précoces et discriminatives sont le gonflement lysosomal et la déplétion en lipides neutres. Une comparaison d'outils d'analyse intégrative a montré le pouvoir discriminatif accru de la suite de biomarqueur entière, par rapport aux réponses individuelles.

This work applies advanced sampling (mark-release-recapture) and comparative approaches addressing the patterns in composition, structure and variability of larval trematode communities in three species of gastropod molluscs (Lymnaea stagnalis, Planorbarius corneus and Radix auricularia) at two nested scales of community organisation in typical Central European eutrophic environments. Hypothesis-testing with the application of null-model analyses, logistic regression modelling and multivariate randomisation techniques, revealed determinants of transmission rates, levels of infection and community structure in freshwater snail hosts in Central Europe and elucidated the mechanisms linking the spatial and temporal environmental variability with the action of complex community assembly rules in freshwater pulmonate snails.

No description available.