Friday 13 July 2012

Ocean Sampling Day‐ Summer solstice 2012‐06‐20 at Faxaflói, Iceland – Matis

Viggó Thór Marteinsson, Eyjólfur Reynisson and Kristinn Guðmundsson

Sampling was performed in Faxaflói, a bay in Southwest-Iceland between the peninsulas of Reykjanes and Snæfellsnes. A small boat was used to go several miles out (Lat: 64° 04,95’ N, Long: 22° 07,50’ V) to the bay at 10:45 o´clock during the summer solstice 2012‐06‐20. Four replicate samples from one sampling station were collected from a depth of 0.2 meter using acid cleaned bucket. Three water samples (1 L) were filtrated instantly on site through four 0.22 μm Sterivex filters (Millipore). Filtering time was around 60 min. The fourth sample was filtered after the other three with total filtration time of 90 min. Filters were sealed and immediately frozen at ‐196 °C in individual zip‐lock bags.

Additional site information:
Air temperature: 12°C
Water temperature surface: 9,6 ̊C
Salinity 3,5%


The skipper (Kristinn) steering the boat to the sampling site - phytoplankton sample – The view to the shore from the sampling sites - Sample filtrated instantly on site through four 0.22 μm Sterivex filters - Filters were sealed and immediately frozen at ‐196 °C – Happy sailors (Eyjó and Viggó)
All the best from Viggó and Eyjó

Wednesday 11 July 2012

Institute of Marine Science (ICM-CSIC). Ocean Sampling Day, Summer Solstice 2012 (20th June).

Sampling was carried out at Blanes Bay Microbial Observatory (BBMO; BBMO is located at the NorthWestern Mediterranean and it is a good example of an oligotrophic (relatively nutrient-poor) coastal ecosystem which is relatively unaffected by human and terrestrial influences. Furthermore, it is one of the sites for which more information exists on the ecology of the Mediterranean planktonic environment, with papers dating back to the 1950's. Finally, it happens to be very close to one marine research laboratory (the CEAB-CSIC in Blanes itself) and not far away from the Institut de Ciències del Mar (ICM-CSIC; ) in Barcelona.

We left the ICM in Barcelona at 9 a.m. on June 20 2012. We arrived at BBMO at 10.30 a.m. The first thing we did was to deploy the CTD to record temperature and salinity profiles and the radiometer to measure light in the water column. We also put the Secchi disc in the water to check the water turbidity. The Secchi disc is a circular and white disc of 30 cm that is mounted on a line, and lowered slowly down in the water. The depth at which the disc is no longer visible is taken as a measure of the transparency of the water. The day was sunny and warm, the sea was flat calm and the temperature of the water was 20 ºC. While we were sampling a couple of friendly jellyfish (Rhizostoma sp.) came to see us.

Regarding to the water samples we collected them from the surface. The seawater was pre-filtered through a 200 µm mesh into 20 liters carboys. These big bottles were transported in the dark, and cold in the case of the samples for RNA, to the ICM in Barcelona. Back in the lab we took samples for chlorophyll a measurements; pigments analysis by HPLC; and inorganic nutrients, organic matter and organic carbon analysis. We also took samples for phytoplankton, picoeukaryotes, bacteria and virus recounts. We measured primary production and bacterial production to study activity of the microorganisms. We extracted DNA and RNA from the picoeukaryote and the bacteria fractions to study the diversity of the microbial population.
Sunrise at BBMO

Friday 6 July 2012

Ocean Sampling Day‐ Summer solstice 2012‐06‐20 - Villefrance

Maria Asplund (PhD Stud), Hans Ohlsson (Tech staff) & Maria Granberg (Assist Prof)

Sampling was performed at the ocean sampling site ”the yellow bouy” (Lat: 58°15’41’’N, Long: 11°27’11’’E) at 13:00 o´clock during the summer solstice 2012‐06‐20. Water was collected from a depth of 1 meter using a water sampler. Water samples were transported to the lab in 4 x 1.3 liter acid cleaned plastic bottles on ice in a thermos within 10 minutes and then filtered instantly. 
(1) 1120, (2)1195 ml , (3) 1180 and (4) 1090 ml water , respectively was run through each of four 0.22 μm Sterivex filters (Millipore). Filtering times were (1) 60 , (2) 65, (3) 68 and (4) 80 min, respectively. Filters were sealed and immediately frozen at ‐80 °C in individual zip‐lock bags. The filters was beige to light brown after the filtering process.

Additional site information
Windspeed: 5 m s‐1
Wind direction : 27
Air temperature: 18 °C
Water temperature 1 m depth: 15 ̊C
Secci depth: 6.5 m
Average Chlorophyll A (0‐2 m): 1.13 μg l‐1
POC: 6.7 ±0.9 (mean ±SE, n=3) mg l‐1

All the best from Maria Asplund and Maria Granberg

VLIZ Ocean Sampling Day 20th June (solstice)

Leaving from Ostend at 8am with our brand new Research vessel Simon Stevin for a visit to a well studied station in the middle of the Belgian Continental Shelf.

We arrived at stat 330 at 11:15 am UTC.

We used niskin bottles to collect our water samples at 3m below surface.

Niskin and CTD are mounted together so we were able to measure a profile for temperature, salinity, turbidity and oxygen saturation during the same deployment.

From the water samples we took 8 replicates:
  • 3 for nutrient analysis and spm
  • 1 for pigment analysis
  • 4 for metagenomic analysis
We filtered the 4 replicates for metagenomic analysis using the Sterivex 0.22µm filters and put 1000mL through each of the filters. This took about half an hour per replicate. (There is definitely room for improving the installation we used. I hope there will not be too much human DNA in the samples☺)

We stored the filters at -20°C in board of the vessel and transferred them to liquid nitrogen on arrival in the harbor.

So far so good, we can start preparing the shipping of the filters!

Thursday 5 July 2012

San Pedro Ocean Time-series Ocean Sampling Day

What better way to appreciate the longest day of the year than waking up at 05:30AM? So, that’s what we did on this year’s summer solstice on June 20. We awoke to an overcast commute down the I405S freeway (one of the busiest in the country) in Los Angeles, CA to San Pedro, CA, but at these hours, of course, traffic is very light. We arrived in time to load the ship with plenty of time to spare for the 7AM departure. Our ship departed from the LA Harbor which is about 25 miles south of downtown LA and the University of Southern California (USC). The LA Harbor is the busiest port in the US and one of the busiest in the world.. Our destination was the San Pedro Ocean Time-series (SPOT) which is about halfway between the mainland in San Pedro, CA and Santa Catalina Island. SPOT is an on-going 12+ year monthly marine microbial time-series. We examine the dynamics of the microbial communities including protistan, bacterial, archaeal, and viral communities. The results show very interesting annually repeating cycles and co-occurrence patterns which may suggest ecological interactions (similar to those easily observed in larger organisms).

With flat seas, it took about an hour to get out to our sampling location. Once on station, we successfully launched our brand new, hopefully very reliable, Rosette/CTD package (THANKS, NSF!!!) down to just above the bottom at 890 meters. This cruise was different than the typical SPOT cruise because we aimed to filter the special samples for Ocean Sampling Day on-board the ship rather than collect and filter back at the lab like we do for the monthly samples. Once the rosette package returned back on board, we collected 2 4L replicates from the 150m, 500m, 890m depths and 4 1L quadruplicates from the chlorophyll maximum (30.2m) and 5m depths. We used peristaltic pumps (7 channels total) to power through all the samples in about an hour. We pre-filtered the samples through an 80um mesh to remove larger organisms (like copepods). The filters were immediately placed on dry-ice. In addition to the OSD samples, we carried out our monthly sampling that included collection of more microbial DNA, bacterial production, and bacterial and viral enumeration.

Upon return to the LA Harbor, we had a minor glitch when we found out that due to homeland security our port was closed, which obviously left us thinking, “Oh no, how long are we going to stuck out here?” However, the captain had a plan and we steamed over to another nearby dock which took an extra 20 minutes. We planed to take cabs over to the vans to return to campus. However, when we arrived at the new dock, the captain found out that our regular dock was open to boats so we returned and went about our business – never a dull moment!

Extra fun stuff: When we looked at the samples from SYBR green epi-fluorescence microscopy we observed that the virus-to-bacteria ratio and Synechococcus abundance were quite high—about 30 and >150,000/mL, respectively.

Diane Kim, Troy Gunderson, and David Needham (L-R) pose with the brand new, NSF-funded CTD/rosette before its first official SPOT deployment.Credit: Victoria Campbell

View of Santa Catalina Island from the San Pedro Ocean time-series through the morning marine layer from R/V Yellowfin. Credit Diane Kim

4 1L replicates from the 5 m depth at the San Pedro Ocean Time-series on ocean sampling day. Note the lighter color on these Sterivex compared with the 5 m samples. Credit: David Needham

4 1L replicates from the chlorophyll maximum at the San Pedro Ocean Time-series on ocean sampling day. Note the darker color on these compared with the 5 m samples. Credit: David Needham

Epifluorescent microscopy images from the 5m depth at the San Pedro Ocean Time-series. Both images were from the same field on a 0.02µm Anodisc filter stained filter stained with SYBR green I. The upper image shows the bacteria and viruses with concentrations greater than 1 and 10 million per mL, respectively. The lower filter shows orange auto-fluorescence of Synechococcus from the same field of view. Credit: David Needham

Cathy Roney happily prepares to collect a sample for Ocean Sampling Day Credit: Diane Kim

Victoria Campbell happily attaches tubing to the Niskin to gently sample the protistan community within. Credit: Diane Kim

Victoria Campbell, Troy Gunderson, and Michael Morando (L-R) sample from the new SPOT CTD/Rosette on Ocean Sampling day Credit:Diane Kim

Jenni Cardell happily takes instruction on how to fill a cubitainers on her first SPOT cruise. Credit Diane Kim

Brian Seegers and Victor Hernando-Morales (L-R) happily sample from Niskin bottles on ocean sampling day and the San Pedro Ocean time-series.
Credit: Diane Kim
David Needham (Fuhrman Lab- USC)

USC Ocean Sampling Day Participants:

Victoria Campbell, Jenni Cardel, Jacob Cram, Troy Gunderson, Victor Hernando-Morales, Diane Kim, Michael Morando, David Needham, Cathy Roney, Brian Seegers

OSD Pilot 2012 - Crete

Monitoring Station: M3A

Report: OSD – HCMR contribution

HCMR Sampling organizers:
Dr. Paraskevi N. Polymenakou (microbiology)
Dr. Giorgos Kotoulas (genetics)
Dr. C. Fragoulis (zooplankton, organizer of the sampling at monitoring station M3A)

People on board:
A. Krystallas, technician
C. Christakis, MSc student

Date of sampling: 20 June 2012
Time of sampling: 12:00 – 13:00 local time
Sampling depths:
2 m, 10 m, 20 m, 50 m, 75 m, 100 m, 120 m.

CTD was deployed on site before sampling to record the vertical profiles of temp, salinity, oxygen, conductivity, chl.

A total of 7 depths were sampled for nutrients, chlorophyll and phytoplankton measurements.

The following five sampling depths were chosen for DNA analysis:
The surface at 2 m to use it as control sample since it is characterized by extremely low concentrations of nutrients and the depths of 50 m, 75 m, 100 m and 120 m where a deep chlorophyll maximum is observed this period (usually this period DCM is observed at a depth of 100 m).

800 ml – 1000 ml of water was filtered through 0.22 um, 47 mm, polycarbonate filters (OSMONICS Inc.), and a total of four replicate samples for each sampling depth. In total we have collected 20 samples (5 sampling depths x 4 replicates). (Unfortunately Millipore Sherivex filters did not arrive on time in order to use them for this sampling.)

On Wednesday 13 of June 2012 (1 week before the OSD-June) a more detailed sampling of the water column was performed. Samples for flow cytometry, chlorophylla, POC, phyto- and zooplankton, and chlorophyll size fraction were collected. In addition sediment samples from the same station using a multi-corer system (enables the collection of samples with undisturbed surface) were also collected.