Tropical cyclone

So the rumors were true. We do have a massive storm in our path, heading straight for us. It is predicted to reach our location in the middle of next week, with 19 meter waves.
The crew doesn't seem to be overly alarmed, our ship is a big and sturdy one, but if we hit a storm like that things (lab equipment included) would inevitably break and we would not be able to run any experiments on board.
Therefore we will attempt to dodge the worst of it by either heading east, somewhat along our intended route (but then miss out on possible long duration station candidates) or head north to seek shelter at Fiji.
The decision will be made within the two coming days, based on our qPCR data and the physical parameters. I think that if we encounter large abundances of UCYN at our our current location (and the next) we will probably head for Fiji.

Last night I ran a qPCR run on water from the short duration station 7, mostly for testing our new standard dilution series aliquots, since the qPCR data wasn't required at that station. I found vast numbers of UCYNB in the top 10 meters of surface water. I think we might have hit a bloom. Perhaps the coming LD candidates will be similar. We're still fairly low on our beloved DDAs though.

Inconsistency and waiting for the CTD

We have now officially entered Fiji waters and crossed into a new time zone, which means that not a single clock on board is showing the actual local time. It also means that I'm now 11 hours ahead of Sweden, which again, feels a bit weird.

It is our 7th short duration station (SD) today, and we are slowly nearing our mid-point (10th of March). The following 2 days will decide where we will have our next long duration station (LD). So today we once again start running the qPCR looking for the UCYN targets.

I don't think I've actually explained what the targets really are. So for those interested, they are small unicellular diazotrophs (nitrogen fixers). Most of them are so small that they can not be identified by any other means than molecular methodologies (e.g. the qPCR). They are currently being subject of some intense research due to their role as diazotrophs but also in their unique characteristics (some arguing that they are in a transitional state between free living and symbiotic, giving an excellent opportunity to study a process similar to the theory of the modern eukaryotic cell, the endosymbiont theory. The reason for this is that some of them have been shown to lack certain genes required for their life history, then being either photoautotrophs or photoheterotrophs. How they still manage without these genes, e.g. for photosystem II essential for photosynthesis, is largely unknown). Some of the other diazotrophs can be seen by the naked eye while others are easily recognised in a microscope.

When we have decided on a location for the second LD, out of three, there will hopefully, be some more consistency in our sampling. At the moment we are trying to save time by sampling whenever we arrive at an intended SD and every single time since LD1 we have had to wait for hours for our first CTD with our water. Tedious, yes.
Anyway, for more detailed information on the cruise plan and goals you can visit the official OUTPACE 2015 webpage at https://outpace.mio.univ-amu.fr/spip.php?article71.

Between rumors of a massive storm, stupid oceanographer community rituals when crossing the dateline and someone spotting a few whales, it is now finally time for sampling.

A day of many things

5/3
So this past day was an intense one. First off, being the first short duration station (SD) in a week, and then so many different things required my attention. I got to bed at 02.30 in the morning and was so tired that I actually missed breakfast, turning off my alarm in the following morning without even noticing. Now it was for once a good thing that the French have their lunch so early (11.00), because I could go straight out of bed, feeling miserable, and down to the mess for brunch, which effectively made me feel less like a hungover teenager.

Firstly, the start of the previous day had us clean out and prepare the lab and qPCR for the SD sampling, which went smooth enough. We even had the time to hit the sunny front deck for an hour.
Secondly, today we also had to decide upon the shipping of samples and equipment back to Sweden after the cruise. I know it is a bit less than a month left but better be prepared and in good time. I would hate to see our samples (which need to be stored and shipped frozen) go to waste because we postponed our decision. We had a bit of a disagreement and communication mishap with our supervisor concerning this, which put me in a foul mood that evening, until I had the time to reply and elaborate on our decision to ship our samples using a recommended French company in Tahiti.
Thirdly, we also had a scientific meeting with one of the research teams on board (this time the physics oceanographers) presenting their data so far, to the rest of the cruise participants.
This took way more time than  I thought it would and since there aren't enough chairs for everyone in the conference room I had to stand (or sit on a sharp edge, numbing my butt) for hours. I quickly got fed up with it.
However, in the middle of the meeting a couple of people from the crew sneaked in and set up a computer with speakers. No one payed them any heed and all of a sudden a silly song in French sounded loudly in the whole room. Startled, everyone started laughing, and they laughed even more when someone in the crew jumped into the room, dressed in the orange immersion suit (environmental life suit, you look really stupid in it) with colorful paper circles taped on his back. He then started dancing in a silly manner, somewhat reminding me of an old friend's poor imitation of Zoidberg from the TV-show Futurama, and hugging everyone in the room. The silly music was still playing by the way.
Apparently today was an annual celebration of some kids TV-show in France. I've never heard of it, but it was a fun distraction from the internal waves measurements in the presentation from the last long duration station (sorry, but you should really get more chairs).
Fourthly, I was contacted by a local newspaper from my home town who asked me to participate in their next issue of "Xtra" which is published once a month (thanks mom). So I had to take a few additional pictures to send them.
Fifthly, SD sampling which was a pain as late as it turned out to be (21.00).

I'm very happy that I despite all the work this day managed to get an hour of calmness on front deck

today, just enjoying the sun and watching the waves roll by as the ship made haste for the next SD.

I know I've said it before, but I can't help being awed by the endless ocean and horizon stretching out around me.
I find it both humbling and inspiring and front deck is a great spot of contemplation. Gazing up at the thin clouds unfolding across the light blue sky like a soothing blanket of white feathers, it made me feel slightly better as the breeze brushed my face and the sun warmed my back.
I felt small but full of possibility. The vast nothingness that is the open ocean is my possibility and I'm right now just a tiny speck in one such ocean of possibilities.
With that thought I once again looked up and tried to point out directions and wondering how far I would be able to see if  I was on the same altitude as the thin clouds. Could I see the curving of the globe? Probably, but not too far in the distance then, even on this bright day, certainly not as far as Sweden. How I miss my fiancée and my kids.

In a hurry towards Fiji

We have now entered a new phase in the cruise. We have left New Caledonia far behind and somewhat entered a  new water mass (at least from a biological perspective). Work-wise it will be the same story though. A "rinse-and-repeat" lab work that we did every day before the first long duration station (LD). The difference now though is that we have a change of schedule. Apparently we are running short on time and consequently are falling behind in the planning, so these coming four days of work, where we would have been contributing with our important data on the UCYN targets abundance in the water column, will instead mean nothing in terms of our qPCR data. The reason for that is that we a rushing over to Fiji waters anyway and we will not stop long enough to consider another LD along that route even though we might hit a bloom of the UCYN.

Sampling will also be done late at night, which is not optimal, and even useless for some of the scientists on board.

The cruise organisers even told us late yesterday evening after we presented our data, that we don't even have to run the qPCR until we get to Fiji unless we really want to. This made me more than a little bit annoyed. How could we be this much behind schedule that they don't even have the time to consider our data. I get the feeling that they have already decided on the spots anyway. This qPCR data is our only real contribution to the cruise and now it's not necessary. I'm more than a little bit curious as to what will happen if we arrive at this intended area, chosen based on satellite surface water imagery, and there are little to no UCYN present.

At least our qPCR run went well. Our data looked good and we had hits on all UCYN targets, the most abundant ones at a magnitude of 10^3. Our initial microscopy sampling also showed decreased abundance of Trichodesmium and finally an increase in the number of DDAs (specifically Rhizosolenia-Richelia). This is good news for me and my colleague. Fingers crossed it will just keep getting better the closer to Fiji we get.

We will probably keep running the qPCR every night for these four days. In the end the data might prove useful and we will at least keep our skills sharp.

Oceanic scientists in need

4/3
So the first long duration station (LD) is officially over and we are well on our way to the next short duration station (SD). We have just rounded the southern tip of Vanatu waters and are heading in a northeasterly direction to get back on route towards Tahiti.
We are estimated to reach the SD at 18.00 local time and me and my colleague will as usual be the first ones to sample of the CTD and get to work on finding any of the UCYN targets using our qPCR. I have a feeling it will be a late workday since our lab procedure usually takes 3,5 hours.
Before that however, we will have to do a test run on our reagents and standards just to make sure they are fine sitting in the freezer for a week. The freezer has proven to be less than reliable regarding the intended temperature of -20 degrees celsius. A higher storage temperature or frequent thawing and freezing will degrade the reagents and standards making them inefficient at best.
After conferring with our supervisor we moved all of it into the -80 degrees celsius freezer instead, but that was no sooner than yesterday.

Apparently a vital instrument called a Winkler for measuring oxygen concentration in the water broke a couple of days ago, so since we are still relatively close to New Caledonia, responsible personnel decided to bring a new one from Nouméa using a helicopter.
This was a procedure which had never been done before so there was a lot of commotion prior to its arrival. Fire fighters were stationed all over the ship in full gear and everyone else, scientists and crew alike (captain excepted), was gathered at back deck for safety reasons. All this just in case anything would go wrong. I know for a fact that the pilot was asked if he was skilled. The answer he gave was that at least he is experienced, meaning that there are no good pilots, they are just either young or old (he being old), which was good enough for everyone involved. I really liked the humble simplicity of that answer.

Anyway, the helicopter arrived and everyone on board started taking pictures of it while it circulated fairly close around the ship. We also realised that they were actually taking pictures of us too. I suppose there was a mutual feeling of respect and awe. I thought it was really cool. The crew in the helicopter was sitting on the edge of the open side door ready to lower down a large crate on deck.
It was all very professional and the actual deliverance was done in less than 10 seconds (the ship was actually still moving too). This was followed by a few more circles around the ship and a wave of goodbye before they disappeared in the horizon, heading south, back to New Caledonia.

Croissant day

It has been two eventful days with a lot of different things going on. Both research wise and not, both immediately concerning my work on board and not.

Either way I might just start out with the things not concerning my research for a change.

As the title says we have a weekly day referred to as croissant day, which happens every sunday. That is also the only way for me to actually know there will be the start of a new week the following day. Anyway, this day we have croissants to eat at every meal of the day, and the bottled red wine (instead of the regular bag-in-box) is served all day too.

As I've previously mentioned I'm not much of a wine drinker but chocolate croissants, yes please! However, I've been told that apparently these aren't good enough for people from Paris. Spoiled French. Whatever. More for me, yay!

The following day marked the end of sampling for this first long duration station (LD), which feels good. It seems that some people have gotten off work earlier though since something reassembling a crude pool turned up on front deck. Beggars can't be choosers, right? So naturally I found myself just hoping they will finish it before we get to Tahiti. Oh, and I also managed to drop a full tray of food on myself at dinner (including a bowl of soup). Don't ask how, because I don't know. I was just in a big hurry to get back to my sampling, typical, and then I had to clean myself up in addition. Previously when someone dropped something there has been a round of applause throughout the cantina, however, after I yelled out a line of less than appropriate words in Swedish at my misfortune, the cantina went dead silent. I'm not sure what would have been worse, the silence or the applause. At least they should all know by now that I'm not American.

The morning of the final day of this LD I was supposed to sample a depth profile from the CTD for use in another, although minor, experiment of mine again involving spiking with 15N2, 13C and filtering for SIMS analysis. But first my bottles needed to be filled, spiked and incubated for 12 hours of light.

I got up at 03.00 to get all set for this experiment, and had decided before hand with the diazotroph team that we would run this experiment together, so once again they would provide me with the 15N2 and 13C while also measuring the bulk nitrogen fixation.

The CTD went down at 03.10. Only me, my colleague, the crane operators and the flying fish being awake. The CTD came back up at 03.45. Still no diazotroph team around. What the hell?

At least there was one more person at the CTD by now who instinctively ran over to one of the cabins of the missing people only to come back with the unimpressive reply that they have their own CTD at 04.00 since they need so much water. God damn it!

Now there are several things wrong with this unfolding early morning so lets just pause there and take it from the beginning.

On the first day of this LD I had a chat with the leading scientist of the diazotroph team and we planned both our schedules so we would be sampling together on the day of the nutrient amendments experiment as well as the depth profile. Mostly since we are both interesting in collaborating now and in the future as well as for me to avoid wasting time and effort running my own bulk samples. On an earlier agreement with my supervisor they are also always providing the 15N2 and 13C since we had some difficulties getting ours delivered to the ship in time.

Anyway, after deciding on our mutual sampling days I went to the cruise organiser to schedule myself and my colleague on the appropriate CTD's during the five days of LD (specifically sampling from the same CTD's as the diazotroph team at two occasions then. At that time there was no mention whatsoever about an extra CTD at 04.00.

The reason is simple. The 03.00 CTD won't be ready for another go until earliest 04.30. The schedule for my CTD also still said that the diazotroph team would be sampling with me.

With that said, we are back at the unimpressive reply which had me, certainly not being a morning person, play out all of my frustration at one go (sorry once more technical personnel), picturing my whole experiment going to hell and getting up at 03.00 for nothing.

Me and my colleague sampled our water anyway and then stored them away in the blue incubators on front deck, for possible use later. 

Sometime after 04.30 part of the diazotroph team merrily entered the lab while whistling and laughing happily. I seized the opportunity and told them of my morning. At least they stopped whistling. They sampled from their CTD and prepared their bottles for incubation using the mooring. To speed things up I helped out as best I could and finally, two and a half hours late, I got my bottles spiked and ready for incubation. After that, karma struck. While we were working on the mooring before grabbing the rest of their samples off the CTD someone had emptied it. I swear it wasn't me.

Break-down of the nutrient amendment experiment

This day marked the end of my major experiment for the long duration stations (LD). After a 36 hour acclimatization, spiking with 15N2 and 13C -bicarb and then incubating for 24 hours it was finally time to filter the bottles with the different treatments.

For my thesis I'm collecting filters for SIMS (secondary ion mass spectrophotometry) analysis. So I want to see if the activity of the nitrogen fixation and growth for the DDAs (diatom - diazotroph association) changes when exposed to non-limiting conditions of certain nutrients.
In my case these nutrients only included iron, DIP (dissolved inorganic phosphorous) and DOP (dissolved organic phosphorous). Of course I also had a control bottle. My ambition was to also include treatments of silicate and vitamin B12, unfortunately there was a last minute change in the availability of these nutrients, so I had to settle with the ones I ran during this LD and a very limited amount of silicate which I will save for an LD where the DDAs are more abundant.

In addition to these SIMS filters I also filtered water for the diazotroph team responsible for measuring the bulk nitrogen fixation which I will use for later comparison. They wanted to specifically preserve filters for DNA and RNA which I will bring back to Sweden after the cruise. Since filtering water for DNA and RNA samples is somewhat my speciality by now, I was happy to assist.
This proved to be a bigger challenge than I had initially thought. There was a lot of multi-tasking involved since the filtration had to be set up for the different bottles and the water for DNA and RNA had to be split up and measured. The different filtrate purposes also required their own special filters and tubes with either a storage medium or nothing, and finally flash freezing in liquid nitrogen or just put in the -20 degrees celsius freezer.
While managing all this, the filtration time for the bottles varied a bit and I had to constantly keep an eye on the bottles and the time taken for each filtration and then get to work on storing the filters in the correct tubes (pre-numbered).
Rockstar filtration! I just missed some metal music in the lab!

All in all it was, as far as I can tell right now at least, a successful experiment. I just hope that I will actually find some DDAs on my filters when it's time to run the SIMS. My hypothesis is that iron will be the biggest stimulant since nitrogen fixation, specifically the nitrogenase complex, requires a lot of iron for its expression. This only takes the symbionts into consideration however. The diatom might need something in addition to iron (which is mainly used in the RUBISCO pigment for carbon fixation) for boosted growth, silicate perhaps.

After this hectic start of the day I treated myself with some quality time on front deck in the hot tropical sun listening to that metal music that I missed in the lab earlier today. The ocean certainly eminates a contagious calmness. I probably watched the gentle waves roll by towards the endless horizon for hours before hitting the gym.

Finally, I have now passed 500 unique views of my blog since I started some 2-3 weeks ago. I'm thrilled about the relatively large interest in my cruise, which far exceeds any expectations I might have had. Thank you all, it is very flattering. In return I give you this picture proving that nature indeed has a sense of humour.