Project description:Monitoring microbial communities can aid in understanding the state of these habitats. Environmental DNA (eDNA) techniques provide efficient and comprehensive monitoring by capturing broader diversity. Besides structural profiling, eDNA methods allow the study of functional profiles, encompassing the genes within the microbial community. In this study, three methodologies were compared for functional profiling of microbial communities in estuarine and coastal sites in the Bay of Biscay. The methodologies included inference from 16S metabarcoding data using Tax4Fun, GeoChip microarrays, and shotgun metagenomics.
Project description:Microplastics represent a growing environmental concern for the oceans due to their potential capability to adsorb different classes of pollutants, thus representing a still unexplored source of exposure for aquatic organisms. In this study polystyrene (PS) microplastics were characterized for their capability to adsorb pyrene (PYR) as model compound for polycyclic aromatic hydrocarbons, and transfer this chemical to filter feeding mussels Mytilus galloprovincialis. Gene expression analyses of Mytilus galloprovincialis exposed to polystyrene (PS) microplastics and to polystyrene contaminated with pyrene (PS-PYR) have been performed trough a DNA microarray platform.
Project description:Pharmaceutical compounds are emerging contaminants in aquatic environment due to their massive use (human and veterinary medicines, agriculture and aquaculture) and a limited removal by waste water treatment plants (WWTPs). In this work, a representative determination of ecotoxicological potential of two different NSAIDs compounds was studied in the sensitive bioindicator marine organism M. Galloprovincialis. Mussels were exposed, under regulated laboratory conditions, to Ketoprofen (KET) and Nimesulide (NIM), dosed alone at the realistic environmental concentration of 0.5µg/L for 14 days. Gene expression analyses of Mytilus galloprovincialis exposed to KET and NIM have been performed through a DNA microarray platform.
Project description:Pharmaceutical compounds are emerging contaminants in aquatic environment due to their massive use (human and veterinary medicines, agriculture and aquaculture) and a limited removal by waste water treatment plants (WWTPs). In this work, a representative determination of ecotoxicological potential of two different NSAIDs compounds was studied in the sensitive bioindicator marine organism M. Galloprovincialis. Mussels were exposed, under regulated laboratory conditions, to Ketoprofen (KET) and Nimesulide (NIM), dosed alone at the realistic environmental concentration of 0.5µg/L for 14 days. Gene expression analyses of Mytilus galloprovincialis exposed to KET and NIM have been performed through a DNA microarray platform. Mussels Mytilus galloprovincialis (5 ± 1 cm shell length) were obtained from a local farm (Numana, Ancona) and acclimatized for 10 days to laboratory conditions with aerated seawater, at 18 ± 1 °C, 37 � salinity, pH 7.5 ± 0.5 and oxygen saturation >94%. Mussels were distributed into three 17 L aquarium and exposed at 0.5µg/L to ketoprofen (KET) and nimesulide (NIM) dosed alone for 14 days. All treatments were compared to control (CTRL) containing 0.00001% of methanol. Water was changed every other day and concentration of molecules were restored. Gene transcription analyses of 12 digestive glands pools (four pools for each treatment composed by 3 digestive glands; CNTR, NIM and KET) were performed using a 8X60K Agilent oligo-DNA microarray platform GPL18667. Microarrays were synthesized in situ using the Agilent non-contact ink-jet technology including default positive and negative controls. Total RNA was isolated using Extract-all (Eurobio) procedure. RNA quality and integrity was controlled on the Agilent bioanalyzer using RNA nanochips and Agilent RNA 6000 nanoreagents (Agilent Technologies, Waldbronn, Germany). RNA concentrations were measured at 260 nm using a ND-1000 spectrophotometer (Nanodrop Technologies) using the conversion factor 1 OD = 40 mg/mL total RNA. Samples were stored at -80°C until further use. Gene expression profiling was performed using an Mytilus galloprovincialis oligo-DNA microarray of 59,971 probes based on single-colour detection (Cyanine-3 only). Microarrays were scanned with Agilent scanner G2565BA at a resolution of 2 microns; all slides were scanned twice at two different sensitivity settings (XDRHi 100% and XDRLo 10%); the scanner software created a unique ID for each pair of XDR scans and saved it to both scan image files. FeatureExtraction v10.7.3.1 used XDR ID to link the pairs of scans together automatically when extracting data. The signal left after all the FE processing steps have been completed is ProcessedSignal that contains the Multiplicatively Detrended, Background-Subtracted Signal.
Project description:We examined gene expression profiling of mussels treated with two chemical mixtures composed by heavy metals or organic compounds, both including genotoxic agents (cadmium, benzo[a]pyrene, 1-nitro/amino-pyrene) and other contaminants (cupper, mercury, fluoranthene, TCDD). For digestive gland one RNA sample from control mussels and one from each treatment group (7 mussels) were competitively hybridized on the MytArray 1.0. Total RNA was reverse-transcribed and labeled using a Cy3-dCTP and Cy5-dCTP direct incorporation. We carried out two separate hybridizations for each treatment conditions. Keywords = mussels Keywords = heavy metals Keywords = organic compaunds Keywords = gene expression profiling Keywords: ordered
2006-10-10 | GSE2183 | GEO
Project description:Aquatic invertebrate metabarcoding with Oxford Nanopore sequencing