Parazitičtí jednorodí (Monogenea) - stále neznámé organismy
VOREL, Jiří, Marie JANKŮJOVÁ, Jan OPPELT, David POTĚŠIL, Lucie JEDLIČKOVÁ, Pavel ROUDNICKÝ, Jana ILGOVÁ, Libor MIKEŠ, Zbyněk ZDRÁHAL, Milan GELNAR and Martin KAŠNÝ. Parazitičtí jednorodí (Monogenea) - stále neznámé organismy (Parasitic monogeneans - still unknown organisms). In Zoologické dny 2018. 2018. ISBN 978-80-87189-24-5. |
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Basic information | |
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Original name | Parazitičtí jednorodí (Monogenea) - stále neznámé organismy |
Name (in English) | Parasitic monogeneans - still unknown organisms |
Authors | VOREL, Jiří (203 Czech Republic, belonging to the institution), Marie JANKŮJOVÁ (203 Czech Republic), Jan OPPELT (203 Czech Republic), David POTĚŠIL (203 Czech Republic), Lucie JEDLIČKOVÁ (203 Czech Republic), Pavel ROUDNICKÝ (203 Czech Republic), Jana ILGOVÁ (703 Slovakia), Libor MIKEŠ (203 Czech Republic), Zbyněk ZDRÁHAL (203 Czech Republic), Milan GELNAR (203 Czech Republic) and Martin KAŠNÝ (203 Czech Republic). |
Edition | Zoologické dny 2018, 2018. |
Other information | |
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Original language | Czech |
Type of outcome | Presentations at conferences |
Field of Study | 10600 1.6 Biological sciences |
Country of publisher | Czech Republic |
Confidentiality degree | is not subject to a state or trade secret |
RIV identification code | RIV/00216224:14310/18:00100816 |
Organization unit | Faculty of Science |
ISBN | 978-80-87189-24-5 |
Keywords (in Czech) | Parazité, Monogenea, Genomika, Transkriptomika, Proteomika |
Keywords in English | Monogenea; parasites; transcriptomics |
Changed by | Changed by: Mgr. Jiří Vorel, Ph.D., učo 376321. Changed: 17/9/2018 15:06. |
Abstract |
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Parazité skupiny Monogenea představují významný soubor organismů, čítající až 25 000 druhů. Jejich prevalence bývá v hospodářských chovech ryb často vysoká a může tak vést k významným ztrátám v populaci hostitele. Ačkoliv se zástupci monogeneí vyskytují kosmopolitně a někteří z nich jsou významnými patogeny, tak znalost biochemické či molekulární podstaty interakce parazit-hostitel je značně omezená. Např. téměř chybí informace o funkčních proteinových molekulách. Doposud byla pouze pro dva tyto organismy (Gyrodactylus salaris a Protopolystoma xenopodis) vygenerována a zpracována celogenomová data a bylo charakterizováno několik málo funkčních proteinů produkovaných zástupci monogeneí. Za účelem prohloubení poznání v této oblasti jsme u druhu Eudiplozoon nipponicum (Monogenea: Diplozoidae) realizovali první komplexní genomickou, transkriptomickou a proteomickou analýzu, která nám umožnila vygenerovat robustní datasety, na jejichž základě bylo možné identifikovat jednotlivé proteinové molekuly potenciálně klíčové pro život tohoto parazita. Pro potřeby genomické analýzy bylo pomocí čtyř sekvenačních platforem (Roche/454 Junior, Illumina HiSeq a MiSeq a Oxford Nanopore MinION) vygenerováno více než 165 mil. čtení a aktuálně probíhá fáze sestavování (assembly) celogenomové informace. V případě analýzy transkriptomu bylo vyprodukováno téměř 150 mil. čtení (Roche/454 FLX Titanium a Illumina MiSeq), 37 062 finálních transkriptů bylo bioinformaticky uspořádáno a z toho 19 539 blíže anotováno. Proteomická data byla získána z exkrečně-sekrečních (ES) produktů z ~100 dospělců pomocí HPLC-MS analýzy a byla porovnána se sestavenými transkripty. Takto bylo identifikováno celkem 1 033 ES proteinů. |
Abstract (in English) |
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Ectoparasitic flatworms from the class Monogenea represent serious fish pathogens and their presence in stocks can lead to significant losses in fish host populations. Despite this fact, the running research has been focused mainly on morphological and phylogenetical characteristics of these worms and the information related to the biochemical and molecular nature of the physiological processes is rather sporadic, as well as the knowledge of the molecules produced by monogeneans and their role in host-parasite interaction. From a large group of Monogenea only a few monogenean protein molecules have been deeply molecularly/biochemically characterized, e.g. peptidases (cathepsins B, D and L) and their inhibitors (cystatins and serpins), thioredoxin and annexin. Up to this date only two monogenean species were deeply sequenced on whole-genomic level. Genomes of Gyrodactylus salaris (Monopisthocotylea: Gyrodactylidae) and Protopolystoma xenopodis (Polyopisthocotylea: Polystomatidae) are currently available in public sequence databases. We would like to change this situation and therefore we adopted Eudiplozoon nipponicum from second large group of Monogenea Polyopisthocotylea (Diplozoidae) as an experimental organism for purpose recognize the important physiological pathways and crucial protein molecules involved in interactions with the fish host. E. nipponicum is a blood-feeding parasite inhabiting gills predominantly of common carp Cyprinus carpio, which is economically very important fish species in the Czech Republic. We started with analyses leaded to generation of E. nipponicum genome, transcriptome and secretome database. For the first E. nipponicum genome draft we used three sequencing techniques. 454/Roche sequencing (Junior platform), Illumina sequencing (MiSeq and HiSeq platforms) and Oxford Nanopore sequencing (MinION platform). 165,871,596 genomic raw reads were generated in total and after quality processing (trimming, removing contaminants, errors correction, etc.), 130,741,241 reads were used for the draft of genome assembly. Currently, the mitochondrial genome was already reconstructed and further bioinformatic evaluation of obtained assembled data is in progress. Generating of transcriptomic/secretomic dataset was based on usage of two sequencing methods (454/Roche and Illumina MiSeq platform) and mass spectrometry analysis (HPLC MS/MS, Orbitrap Elite Hybrid spectrometer). Total RNA, in the form of cDNA, was sequenced and raw reads were processed and assembled using specific bioinformatic tools. Based on the statistical evaluation and post-filtering steps, final transcriptome database contains 37,062 contaminant-free transcripts with corresponding translated NUMBER proteins. During annotation step, 19,539 (52.7 %) transcripts were homologous to some record in selected public sequence databases and 18,556 (50.1 %) of them were linked with sequences related to organisms only in phylum Platyhelminthes deposited in UniProtKB/TrEMBL database. Excretory-secretory products (ESP), protein compounds present in parasite secretome, were obtained from ~100 living adults and analysis of secretome peptide mixture revealed 1,033 proteins. |
Links | |
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GBP505/12/G112, research and development project | Name: ECIP - Evropské centrum ichtyoparazitologie |
Investor: Czech Science Foundation | |
MUNI/A/0816/2017, interní kód MU | Name: Výzkum ekologicko-evolučních vztahů bezobratlých živočichů (Acronym: EKOLINKS) |
Investor: Masaryk University, Category A |
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