Project description:We aimed to exemplify early and late transcriptional response of M. galloprovincialis to live Vibrio cells. Hemolymph was collected at 3 and 48 hours after the injection of 10 to 7 cells Vibrio splendidus LGP32 into the posterior adductor muscle. Hemolymph samples were similarly collected from paired control mussels injected with PBS-NaCl. The purified RNAs were successfully amplified, labelled and competitively hybridized to the new mussel oligoarray Immunochip 1.0.
Project description:We aimed to exemplify early and late transcriptional response of M. galloprovincialis to live Vibrio cells. Hemolymph was collected at 3 and 48 hours after the injection of 10 to 7 cells Vibrio splendidus LGP32 into the posterior adductor muscle. Hemolymph samples were similarly collected from paired control mussels injected with PBS-NaCl. The purified RNAs were successfully amplified, labelled and competitively hybridized to the new mussel oligoarray Immunochip 1.0. After acclimatization, groups of 40 farmed mussels from the Venice lagoon (Italy) were injected either with 10 to 7 cells of Vibrio splendidus LGP32 or with NaCl-enriched PBS. One ml of hemolymph was withdrawn at 3 h and 48 h post-injection from each control or treated mussel. Two RNA pools (N=10) per time point were composed from the treated mussels, processed and competitively hybridized in dye-swap combination (Cy3 / Cy5 aRNAs) on the same Immunochip slide against time-paired control RNA pools (N=40). Since each Immunochip array contains 4 replicates per probe, the dye-swap testing yielded a total of 16 expression values per probe per time-point.
Project description:Mussels (Mytilus galloprovincialis) were exposed during 24 hours to a waterborne infection with 10E8 CFU/ml Vibrio splendidus (reference strain LGP32) in the tank water. Five biological replicates were used for each infected and control conditions.
Project description:Mussels (Mytilus galloprovincialis) were exposed during 7 and 28 days in seawater (control), seawater + acetone (solvent control, SC), and 10 micrograms per Litre of tris(1,3-dichloro-2-propyl) phosphate (TDCPP). TDCPP was added from a stock solution prepared in acetone, the volume added being 100 µL per 30 L aquaria. The same volume of acetone was added to SC aquaria. Mussel density was 20 mussels per each 30 L aquaria at the beginning of the experiment and varied between 15 ‒ 20 mussels/30 L aquaria during the 28 days exposure period. Exposure conditions were the following: T = 15.6 ± 0.7 ºC, S = 35.5 ± 0.5 ppt, pH = 7.9 ± 0.1, O2 = 7.9 ± 0.6 mg L-1 and 10:14h light:dark photoperiod. Water was renewed twice per week and mussels were fed before every water renewal with a mixture of phytoplankton representing 1% of mussel tissue dry weight.
