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| Status |
Public on Feb 22, 2016 |
| Title |
Pollutants bioavailability and toxicological risk from NSAIDs to marine mussels |
| Organism |
Mytilus galloprovincialis |
| Experiment type |
Expression profiling by array
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| Summary |
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.
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| Overall design |
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.
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| Contributor(s) |
Milan M, Bargelloni L, Mezzelani M, Regoli F |
| Citation(s) |
25637744 |
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| Submission date |
Mar 17, 2015 |
| Last update date |
May 24, 2016 |
| Contact name |
Massimo Milan |
| E-mail(s) |
[email protected]
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| Phone |
+39 0498272941
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| Organization name |
University of Padova
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| Department |
Dept. of Department of Public Health, Comparative Pathology,and Veterinary Hygiene
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| Street address |
Viale dell'Università 16
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| City |
Legnaro |
| State/province |
PD |
| ZIP/Postal code |
35020 |
| Country |
Italy |
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| Platforms (1) |
| GPL18667 |
Agilent-044345 UniversityPadova_Mytilus galloprovincialis_8x60K |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA278647 |
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