Molekulární adaptace neurotropních a viscerálních schistosom během infekce ptačího hostitele

Genus Schistosoma is one the most studied group of helminths due to the importance of several representatives in terms of veterinary and human health. The advent of the modern sequencing technologies, as well as the increasing computational capacities, enabled large-scale screening of nucleic acids and thus deep exploration of complex transcriptome and genome information. To date the main attention of leading molecular parasitological "Schistosoma" research teams was focused on serious human pathogens Schistosoma mansoni, Schistosoma japonicum and Schistosoma haematobium. In the term of molecular/biochemical research, the other schistosomatids were mostly neglected and general knowledge was limited to characterization of particular genes/proteins without further link to biological functional complex. Presented thesis summarises the first large-scale insights into the molecular basis of biological principles of two bird schistosomes Trichobilharzia regenti and Trichobilharzia szidati during their invasion of the definitive avian host. While T. szidati uses the "classical" visceral way of migration - bloodstream and lungs (same as human schistosomes), T. regenti, migrates trough the peripheral nerves and spinal cord. Neurotropic migration is unique among schistosomes and it is also extremely rare...Rod schistosoma je jednou z nejvíce studovaných skupin helmintů vzhledem k významu některých zástupců z hlediska humánní a veterinární medicíny. Rozvoj moderních sekvenačních technologií a se stále se zdokonalující výpočetní technika umožnily globální pohled na molekulární biologii schistosom za použití genomiky a transkriptomiky. Dosud byla pozornost předních vědeckých týmů zabývajících se schistosomózou zaměřena na významné lidské patogeny Schistosoma mansoni, Schistosoma japonicum a Schistosoma haematobium. Ostatní zástupci čeledi Schistosomatidae byli z hlediska molekulární biologie/biochemie většinou opomíjeni a výzkum byl zaměřen především na jednotlivé geny/molekuly, bez celkového přehledu. Předkládaná práce prezentuje komplexní transkriptomickou analýzu ptačích schistosom - neurotropní Trichobilharzia regenti a viscerální Trichobilharzia szidati během nákazy kachny jakožto definitivního hostitele. Zatímco T. szidati využívá běžnou viscerální migraci (stejně jako lidské schistosomy), T. regenti využívá k migraci periferní nervy a centrální nervovou soustavu. Způsob neurotropní migrace je mezi schistosomami unikátní a také velmi vzácný v rámci všech helmintů. Práce se zaměřuje na molekulární mechanismy spojené s viscerální a neurotropní životní strategií obou druhů pomocí transkriptomické...Department of ParasitologyKatedra parazitologiePřírodovědecká fakultaFaculty of Scienc

Dispersal and larval hosts of the zigzag sawfly Aproceros leucopoda (Hymenoptera) in Slovakia, Central Europe

Background and purposeThe zigzag elm sawfly, Aproceros leucopoda (Hymenoptera), originating from Asia is being considered as one of the major pests of elm trees (Ulmus spp.) within Europe where it was first recorded in 2003. Recent investigations document the fast spread of this invasive species in many European countries. In Slovakia, A. leucopoda was first recorded in 2009 but its occurrence here has not been reported since 2009. Herein, we summarize the data from the two consecutive years about the current distribution of A. leucopoda and its hosts in Slovakia, Central Europe, focusing on infestation of elm trees by this successful invader in different altitudes.Material and methodsIn 2014 and 2015 the investigations of A. leucopoda were conducted at 101 different plots across the country. At each plot, characteristic symptoms of leaf damage, occurrence of larvae, cocoons or adults were recorded along with the geographical coordinates and the altitude, and the host trees of A. leucopoda were identified at the species level. The distribution map of A. leucopoda was drawn using the field data. The infestation of elm trees along the altitudinal gradient was analyzed with Kruskal-Wallis non-parametric analysis of variance followed by post-hoc Dunn\u27s test for multiple comparisons.Results and conclusionsThe zigzag elm sawfly A. leucopoda was observed at 31 plots (30.7%, n = 101). In elm trees examined, generally, a low level of infestation by this pest was observed. A. leucopoda was found at plots between 107 and 701 m a.s.l. The result show that the infestation level of elm trees was decreasing with increasing altitude. Although we had only a small sample of data, we assume that the altitude has a significant effect on distribution of this pest. The field elm (Ulmus minor Mill.) was the most frequently utilized host. Data collected document the establishment of this successful invader in Slovakia.</p

Molecular adaptations of neurotropic and visceral bird schistosomes during the infection of the avian definitive host

Genus Schistosoma is one the most studied group of helminths due to the importance of several representatives in terms of veterinary and human health. The advent of the modern sequencing technologies, as well as the increasing computational capacities, enabled large-scale screening of nucleic acids and thus deep exploration of complex transcriptome and genome information. To date the main attention of leading molecular parasitological "Schistosoma" research teams was focused on serious human pathogens Schistosoma mansoni, Schistosoma japonicum and Schistosoma haematobium. In the term of molecular/biochemical research, the other schistosomatids were mostly neglected and general knowledge was limited to characterization of particular genes/proteins without further link to biological functional complex. Presented thesis summarises the first large-scale insights into the molecular basis of biological principles of two bird schistosomes Trichobilharzia regenti and Trichobilharzia szidati during their invasion of the definitive avian host. While T. szidati uses the "classical" visceral way of migration - bloodstream and lungs (same as human schistosomes), T. regenti, migrates trough the peripheral nerves and spinal cord. Neurotropic migration is unique among schistosomes and it is also extremely rare..

Species of the genus Radix in transmission of trematodes in the Czech Republic

Snails of the genus Radix play an important role in the transmission of many species of trematodes which can represent dangerous parasites of animals and humans. Taxonomical situation within the genus Radix is unclear. It is caused by morphological plasticity of the shells of particular species. In the past the shell shape was the most frequently used determination criterion for new species description. Due to new data brought by molecular taxonomy, validity of some species became doubtful. In our contribution, modern methods (phylogenetic analysis based on gene sequences) and classical approaches (determination by conchological features and anatomy of reproductive organs) were used for determination of snails belonging to the genus Radix. For phylogenetic analysis two genes were characterized (mitochondrial 16S rDNA and nuclear ITS2 rDNA); in the samples 5 species of the genus Radix (R. auricularia, R. labiata, R. lagotis, R. peregra a R. ampla) were confirmed. A thorough morphological determination was based on 11 conchological characters which allowed to identify all selected snails; conchological and molecular identifications were in agreement. For determination by means of reproductive organ morphology, shape and position of bursa copulatrix and its duct were used. According to these criteria,..

Additional file 2: Table S2. of Complete mitochondrial genome of the giant liver fluke Fascioloides magna (Digenea: Fasciolidae) and its comparison with selected trematodes

Comparison of A + T content of mitochondrial genomes of Fascioloides magna (Fm), Fasciola hepatica (Fh) and Fasciola gigantica (Fg). (DOCX 20 kb

Preconcentration of hydride forming elements in quartz trap in connection with ICP-MS

Charles Uviversity in Prague Faculty of Science Department of Analytical Chemistry Student: Miroslav Kašpar Supervisor: RNDr. Tomáš Matoušek, Ph.D. Name of Thesis: Preconcentration of hydride forming elements in quartz trap with ICP-MS detection The aim of this thesis are pilot experiments employing quartz trap (QT) for preconcentration of hydride forming elements and their subsequent detection by inductively coupled plasma mass spectrometry (HG-QT-ICP-MS). Arsenic and antimony were chosen as model analytes. Preconcentration efficiency on quartz surface was studied in a trap, which was integrated into an interface between spray chamber and plasma torch. We have identified a critical loss of analyte during transportation, instability of the plasma due to sudden change of gas composition, poor repeatability and very quick release of Sb from the trap resulting in difficult quantification of Sb signal. Despite the efforts to resolve these experimental problems, so far we were unable to achieve improvements in analytical parameters compared to standard analytical methods. Keywords: Preconcentration, quartz trap, mass spectrometry with inductively coupled plasma, hydride generation, arsenic, antimon

Isoforms of Cathepsin B1 in Neurotropic Schistosomula of Trichobilharzia regenti Differ in Substrate Preferences and a Highly Expressed Catalytically Inactive Paralog Binds Cystatin.

Schistosomula (the post-infective stages) of the neurotropic schistosome Trichobilharzia regenti possess multiple isoforms of cathepsin B1 peptidase (TrCB1.1-TrCB1.6) with involvement in nutrient digestion. The comparison of substrate preferences of TrCB1.1 and TrCB1.4 showed that TrCB1.4 had a very narrow substrate specificity and after processing it was less effective toward protein substrates when compared to TrCB1.1. Self-processing of both isoforms could be facilitated by sulfated polysaccharides due to a specific binding motif in the pro-sequence. Trans-activation by heterologous enzymes was also successfully employed. Expression profiling revealed a high level of transcription of genes encoding the enzymatically inactive paralogs TrCB1.5 and TrCB1.6. The transcription level of TrCB1.6 was comparable with that of TrCB1.1 and TrCB1.2, the most abundant active isoforms. Recombinant TrCB1.6wt, a wild type paralog with a Cys29-to-Gly substitution in the active site that renders the enzyme inactive, was processed by the active TrCB1 forms and by an asparaginyl endopeptidase. Although TrCB1.6wt lacked hydrolytic activity, endopeptidase, but not dipeptidase, activity could be restored by mutating Gly29 to Cys29. The lack of exopeptidase activity may be due to other mutations, such as His110-to-Asn in the occluding loop and Asp224-to-Gly in the main body of the mature TrCB1.6, which do not occur in the active isoforms TrCB1.1 and TrCB1.4 with exopeptidase activity. The catalytically active enzymes and the inactive TrCB1.6 paralog formed complexes with chicken cystatin, thus supporting experimentally the hypothesis that inactive paralogs could potentially regulate the activity of the active forms or protect them from being inhibited by host inhibitors. The effect on cell viability and nitric oxide production by selected immune cells observed for TrCB1.1 was not confirmed for TrCB1.6. We show here that the active isoforms of TrCB1 have different affinities for peptide substrates thereby facilitating diversity in protein-derived nutrition for the parasite. The inactive paralogs are unexpectedly highly expressed and one of them retains the ability to bind cystatins, likely due to specific mutations in the occluding loop and the enzyme body. This suggests a role in sequestration of inhibitors and protection of active cysteine peptidases