Dunstaffange

Achievements for each user project

Covadonga Orejas Saco del Valle

Field work
A research cruise was carried out (3- 6.05.2010) on the RV Calanus in the Mingulay Reef complex. Sampling was very successful and up to 12 grabs were collected, each of them with colonies of Lophelia pertusa to develop the experiments.

Feeding short term experiments
Three different treatments (T) were applied:

T1. Addition of an algal culture of Skeletonema costatum. Due to the shallow depths of the Mingulay Lophelia reefs, it is likely that Skeletonema could be a potential food source for the corals.
T2. Addition of a mixed algal culture (4 algae strains). This mixture gave different sizes and shapes in order to look for possible preferences (if any) in relation to different sizes and shapes.
Both treatments were tested at 2 different current speeds (5 and 10 cm/s). Samples from each experiment were preserved in formaldehyde 4% and frozen by -80ºC and in lugol.
T3. Natural fresh zooplankton collected in the vicinity of the SAMS. This was a good opportunity to look for capture rates of Lophelia pertusa in freshly collected zooplankton. This was tested under 3 peeds: 2, 5 and 10 cm/s. Samples from each experiment were preserved in formaldehyde 4%.

Results from the T3 have been processed and are very promising. These results will be presented in December in the ISRS congress in Holland (http://www.isrs2010.org)

Polyp behaviour experiments. Flume
The flume was used to test the polyp behaviour of L. pertusa under a current of 2cm/s and different temperatures (8ºC ->
14ºC). 3 trials were run at 8ºC (field temperature at Mingulay), and 3 were conducted with 8ºC as a start temperature gradually increasing over several hours until reaching 14ºC (extreme temperature for Lophelia).

All experiments have been successfully developed, the SAMS facilities as well as the support personal (researchers and techinicians make our work very pleasant and succesful).

Lipid Biomarker characterisation
To characterise the lipid biomarkers a long term experiment (6 months) will be set up at SAMS. Two different diets (algae: S. costatum; zooplankton: Artemia) and an unfed control will be established. Start of the experiment is expected for September 2010. Samples will be taken over the 6 months, respiration measurements will be carried out with each nubbin and afterwards nubbins will be buoyant weighed and then preserved by -80ºC for further analyses.
As preparation for the long-term experiments, respiration trials were conducted. The aim was to ascertain the duration of respiration experiments (during which pO2 fell from 100% to 80%), and the shortest duration of acclimation. Lophelia nubbins were placed in specially built chambers overnight andrespiration rates measured the following day using electrodes. Further trials are being conducted with optodes to enable a protocol to be developed.

RU-JIN Huang

A 70 L galss/Teflon chamber was set up and demostrated to be feasible for the proposed seaweed related studies during the execution of the project. Some preliminary studies of seaweeds (i.e., Laminaria digitata, Ascophyllum nodosum and Fucus vesiculosus) showed new aerosol particle formation derived from iodine release from the investigated seaweeds. Unfortunatelly, however, we encountered two major obstacles during this visit to SAMS: (1) we placed the order of 2 cylinders of synthetic air that are necessary for our seaweed chamber before my visit, however, when I arrived at Oban we found this purchase was cancelled by BOC since Oban is a remote site and the transport is not convenient as stated by BOC. We therefore spent one more week to get the synthetic air; (2) the ozone generator/monitor failed to work during the visit although we tried to repair it. These problems resulted in an unsatisfactory output. However, we intend to revisit Oban to continue this project with the support from German Research Foundation (Deutsche Forschungsgemeinschaft, DFG).

Gabriele M. König

Achievement: cultured 22 strains in the scale of 1 L culture and obtained good cell mass in 20 strains with no growth in 2 out of 22 strains. The cells from 20 strains were subjected to methanol extraction. The natural product profiles of these extracts were assessed by LC-high resolution MS. 11 out of 20 strains produced new natural products which haven´t been reported before. We will pursue to isolate and characterize these natural products in the near future.

Impact: if new natural products can be identified, the results will hold potential for future medicinal application.
Difficulties: It is difficult for us to continue the project without any funds to culture the strain again for natural product isolation and characterization if we have positive
hit.

Antonios Zambounis

The elicitors were chosen to induce or mimick presumptive defense transduction pathways. We selected candidate stress markers among genes known to be induced in similar or other stress conditions, designed and validated specific PCR primers to monitor their expression profile with qPCR. We conducted elicitor treatment on Ectocarpus cultures, along with inoculations with the parasite Eurychasma dicksonii.
Biological material required for the two following follow-up experiments was successfully generated and harvested:
1)a preliminary qPCR transcriptional analysis of stress gene marker
2) a transcriptome-wide SOLEXA-based study of Ectocarpus defenses against biotic and abiotic stress.

 

Hanna Laakkonen

The immediate purpose of the visit was to collect material of several marine organisms for a comparative phylogeographic study of amphi-boreal marine fauna, distributed both in the Atlantic and Pacific oceans. The general objective of the project Is to use molecular data to get an overall view of trans-Arctic connections of marine biota, on different time scales.
Study material was successfully collected of 12 invertebrate species relevant for the project.

Alasdair M. COOK

We screened for six sulfonates in 38 samples of red, brown or green algae from 3 dives. The sulfonates were isethionate, sulfoacetate, taurine, cysteate, homotaurine and sulfopropanediol.
Neither cysteate nor sulfoacetate was found and traces only of isethionate (1) and homotaurine (5) were suspected. The presence of taurine (11) and especially sulfopropanediol (14) was indicated: these latter samples are being subject to analysis by MALDI-TOF-MS in order to evaluate the tentative identifications by HPLC (taurine) or enzymic analyses (sulfopropanediol). Two other amino acids were detected, glutamate and alanine. These two compounds were found in high amounts in Palmaria palmata, which, alone, contained significant amounts of both taurine and sulfopropanediol, and possibly homotaurine. This seaweed has been used for millennia as a foodstuff.
The samples from CCAP cultures, which were collected and stored but not processed, will be examined in July 2010.
Initial experiments showed that the sensitivity of the methods was low, so an improved extraction technique was developed. The enzymatic assay of isethionate did not work in practice, but our IC methodology was improved to enable satisfactory analyses. The enzymatic assay of sulfopropanediol was subject to interference, but the assay allowed potential positives to be identified for mass-spectrometric analyses.
The data confirm the presence of sulfonates in native algae, so we feel it worthwhile to culture genome-sequenced algae, with the aim of identifying a suitable organism in which to elucidate the pathway(s) of sulfoglycolysis. The function of these sulfonates, e.g. sulfopropanediol, has also to be established.