MassIVE MSV000093578

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Interactions between sea snails and trematodes: analysis of metabolomes

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Description

This work is devoted to the analysis of parasite-host interactions at the metabolome level. We analyzed the metabolome of two species of molluscs, Littorina saxatilis and L. obtusata, healthy and infected with trematodes Microphallus pygmaeus. For analysis of the mollusc metabolome, snail soft tissues (without hepatopancreas and operculum) were separated and fixed individually in 100% methanol. Five to eight biological replicates per each group of comparison were collected (53 samples in total). To analyse the parasite metabolome, we used daughter sporocysts with metacercariae of M. pygmaeus purified from host tissues in filtered sea water (0.2 mkm filters, Merck). Parasite tissues were fixed in 100% methanol. Six to eight biological replicates per each group of comparison were collected (28 samples in total). The prepared material was fixed and then frozen initially at -20oC and later at -80oC until use. Metabolomic data were obtained using nontargeted GC-MS (gas-chromatography mass spectrometry)-based profiling of the trimethylsilyl derivatives. Shortly: the frozen tissues were homogenised, centrifuged, afterwards the supernatants were vacuum-dried and dissolved in pyridine (Merck) containing 1 mg/ml of an internal standard (nC23, Sigma-Aldrich). The silylation agent (N,O-bis(trimethylsilyl)-trifluoroacetamide with 1% trimethylsilyl chloride, Sigma-Aldrich) was added to each sample before the analysis. The analysis was carried out on a gas chromatograph with a time-of-flight mass spectrometer Pegasus 4D GCxGC-TOF MS (Leco). Chromatographic separation was performed in Zorbax DB-5 columns ((5%-phenyl)-methylpolysiloxane; length 30 m, inner diameter 0.25 mm, film thickness 0.25 mkm; Agilent Technologies). Analysis method: evaporator temperature 250oC; starting temperature 70oC; gradient 6oC/min; final temperature 320oC; helium carrier gas, flow rate 1 ml/min. Mass spectrometry: detection frequency 10 spectra/sec in the mass range 50-800Da. [doi:10.25345/C5XD0R80Q] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: molecular parasitology, host-parasite interactions, metabolomics, GC-MS

Contact

Principal Investigators:
(in alphabetical order) 		Egor A. Repkin, Saint-Petersburg State University, Russia
Submitting User: egor_repkin

Publications

Egor A. Repkin, Elizaveta R. Gafarova, Marina A. Varfolomeeva, Dmitrii S. Kurjachii, Dmitrii E. Polev, Alexei L. Shavarda, Georgiy P. Maslakov, Roman I. Mullakhmetov, Ekaterina V. Zubova, Timur B. Bariev, Andrei I. Granovitch, Arina L. Maltseva.
Littorina snails and Microphallus trematodes: Diverse consequences of the trematode-induced metabolic shifts.
Repkin, E.A., Gafarova, E.R., Varfolomeeva, M.A. et al. Littorina snails and Microphallus trematodes: Diverse consequences of the trematode-induced metabolic shifts. Parasitol Res 123, 229 (2024). https://doi.org/10.1007/s00436-024-08244-8.

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