2018 Day 13. Back home

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25 May 2018

A bit late this update of our last day in the #ChernobylFrogs18 campaign, but that’s what happens when you arrive home from Ukraine to find a general failure of internet in your neighborhood that will last for a week…

This was the final day of our 2018 campaign in Ukraine investigating the effects of the chronic exposure to the radiation released by the Chernobyl accident on the amphibians.

It was time to go for the last time to our lab at Slavutych, check the dry shipper levels once more, and put everything in the van that will transport us to Kiev airport. This time, we travelled with our expert in bureaucracy from the Chornobyl Centre, to help us with the export and flight permits. This time the trip was not as shaky as the previous one, a much better road that in about two hours leave us to the airport.

Once there, the first stop was with the airport veterinary, to check that all our samples were in order, and we had all the permits needed to fly with them. It took about one hour of waiting but there was no problem at the end. Next stop was at the oversized luggage for Jean-Marc, the first to fly and the one carrying the dry shipper. Again, all fine, it as last year… However, this lack of problems did not last until the end. Soon by realize that Jean-Marc flight to Frankfurt was delayed. So much delayed that we star fearing about his possibilities to make the connection to Marseille (only 1:05h between the two flights). At the end, the flight took off with about one hour of delay. To me, it was clear that it was not possible for him (even less for the oversized shipper) to make the connection. And, even worse, that flight was the last one to Marseille that day, with a strike of traffic controllers organized for the following day. Things did not look good. If something happens to the shipper, all the work of these days could be at risk, most of our samples lost.

With this on mind, it was time for Pablo and me to get our flight back to Sweden. We arrived even before scheduled, just to know that somehow (I still don’t understand how…) Jean-Marc and the shipper were in Marseille. No problems, all good. All happy!!!

Now, it is time for us to organize this material, all the data and samples collected during this intensive campaign. Soon, Jean-Marc will fly here with part of the samples that we will analize here. Others will stay in France for other different analyses. Time also to rest a bit.

Next, and final post of the campaign will be a summary of all our work during these weeks in Ukraine, a great time spent with our amazing Ukrainian colleagues, and with a very successful result.

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2018 Day 12. Packing time

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24 May 2018

Our last day of work in Ukraine for this season. After almost two weeks working here, it was time for us to finish sampling the last frogs and pack all our equipment. We are ready now for going back to Kiev and to our research centers in Sweden and France. All the samples collected here. Together with the ones collected in 2016 and 2017, they will keep us busy for a very long time.

During the morning, we worked in the lab as usual, sampling the 9 frogs collected the previous night in a clean locality close to Slavutych. We used one of these frogs for a full sampling of tissues with the idea of joining the two from last day as references for our genomic and transcriptomic studies. We also run our last blood samples on our analyzer, for a total of about 100 samples during the campaign, including some in duplicate to test the reliability of the machine. This has been something new for this year, and something that has worked incredibly well!!

Everything was finished for lunchtime. All frogs sampled, all material clean, all vials stored in our dry shipper full of liquid nitrogen for now. Back from lunch, it was time to carefully pack all our sampling material, the camera, iSTAT, boxes of non freeze samples (sperm, bone for skeletochronology) and blood smears for immunological studies. Some of these things will go with Pablo and me back to Uppsala, and some with Jean-Marc to France, together with the shipper. In a few days, Jean-Marc will travel with quite a few of these frozen samples to Uppsala, so we can start to work with them quickly.

Everything was ready pretty soon. It was also time to announce the winner of our “Chernobyl Frogs 2018: The Game”. It has been funny, especially during the last days, to see how close or far people was for the real number of frogs. Many, including ourselves, were too optimistic suggesting numbers well over 150 frogs… At the end, the total number of frogs collected in the trip was 109. And the winner, Claire Keeley, predicting 107, really, really close!! Also very close were Martina Ferraguti (104) and Rafael Gutierrez (103), with just a bit too few, and Jesús Orizaola (117) and Katja Guschanski (117), with just a bit too many. Well done everyone!! And thanks for participating!! The photo will be going to Claire very soon!!

Time to leave the Slavutych lab for the last time. Tomorrow we will pass by here to put all this material in our van, and drive to Kiev-Boryspol airport. Jean-Marc will be the first one departing at 5:30 PM, then it will be time for Pablo and me to fly to Sweden at 8:30 PM. The idea is to leave Slavutych with plenty of time, we have ahead bumpy roads, possible traffic jams, airport bureaucracy for exporting the samples, and we need to be ready for the usual complications with our dry shipper (hopefully not as many as last year). Luckily, we will have people from the Chornobyl Center helping us with the paperwork. Fingers crossed for good trips for us, and our samples!! One more day here, the last of our work in Ukraine. Time to get back home with the family, and back to the working routines. Tomorrow, last daily dispatch from the field!!

2018 Day 11. Last night in the field

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23 May 2018

That’s all. Last night was our last time in the ponds collecting frogs for our #ChernobylFrogs18 campaign. And was, again, a successful night. Every single night, except the first one, we have managed to collect frogs. That’s quite remarkable. And although we have caught not as many frogs as we wanted (as always), we can say now that the trip has been extremely successful!!

We started the day in the lab, sampling our frogs from last night, the first outside the Chernobyl Exclusion Zone. These ten frogs are really important as they will represent our reference for individuals living in areas not affected by radiation. They are our baseline to which to compare the individuals from high, medium and low contamination areas that we collected the previous days, and the previous years since the start of the project in 2016.

It took us quite some time to process these frogs, in particular the last two ones for which we took additional samples to be used as references in our genomic and transcriptomic studies. With these frogs we have sampled already 100 individuals, about 1400 criovials, and 88 successful blood profiles with the iSTAT analyzer. This will keep us busy from quite some time, no doubt…

On the afternoon, we moved to an area of flooded meadows near Slavutych trying to catch the last frog from the campaign. This time we did not use our trusty Chinese car (“the tank”) but a legend of Chernobyl research, a car that has accumulated more than 800.000 kilometers (“the crocodile”, as Kat Raines named). That’s more than going to the moon and back!!!

Once again, we arrived with plenty of time to the field, a really nice place full of flowers and birds, and with plenty of fire-bellied toads calling in the distance. It was also time to be surrounded one more night by masses and masses of mosquitoes. But it allows us to see the area and plenty of beautiful iris!!

We walked and waited, and walked and waited… Until, with the last light of the day disappearing, we heard the first treefrog call. Not too may it seemed, not too active, but probably enough to use it as our final locality and final frogs of the year. We moved into the ponds, and indeed it was clear that we will need to move up and down, here and there, if we wanted to have some frogs… And that we did. It took us almost two hours to catch some frogs, until all calling activity stopped. That’s it, these are the last frogs of the trip.

Same as in the previous night, these frogs live in a clean area, a place that was not affected by the radioactive fallout in 1986, and will be part of our baseline, our reference to compare them with the Chernobyl frogs. Really precious frogs indeed!!!

By the way, for the people participating in our “Chernobyl Frogs Game”, how many frogs did we caught last night? What is it the grand total? Well, we will just keep a bit the suspense, until the next post, ok?

The plan for today is to sample these frogs during the day here in Slavutych, pack all our field and lab stuff, and get ready to travel back to Kiev on Friday morning for our flights back home. People here start to be really tired of the hard work and the constant fiel-lab routine every day. Time to get back and get some rest.

Have a nice day!!

2018 Day 10. Clean frogs

22 May 2018

Our first day outside the Chernobyl Exclusion Zone was our chance for collecting frogs from areas never contaminated by the nuclear accident in 1986. And sure we did. During the night, we managed to collect 10 more frogs in a clean area East of Slavutych. This puts our total, with one last night remaining, in 100 frogs collected from 7 different localities. Really, really, good results!!

During the day, with no frogs to work with at the lab, we set up all our things in our new lab for this part of the trip, the Radioecology Laboratory of the Chornobyl Centre. We organized all our tubes, dry shipper, cameras, and sampling material in our new space, ready and waiting for new frogs. This lab is really great, an amazing facility with plenty of space for doing our work.

We also used our time to visit several locations that look promising for having treefrogs South of Slavutych, in clean areas, with no radioactive contamination. We had a look at two different places, both really nice meadows with no human activity either, full of grassland birds and good conditions for having frogs, even one with a small beaver dam. We will return here today on our last day of frogging in Ukraine!!

During the night, together again with Sergey and Zenja, and the trusty Chinese 4×4 car, we moved to the East-Southeast of Slavutych trying to find treefrogs in areas never affected by the radioactive fallout. After driving for about 1h30, we arrived a two promising places, with plenty of water. That much water that we had to abandon one since we were not able to continue with our car.

We moved to a second location just to realize that I had forgot our head lamps in the lab, a disaster. Thanks to my spare headlamp and one from Jean-Marc we finally managed to have some light for looking for frogs, although not that much. Next, I saw that one of the waders we had had a hole in one leg, 100% guaranteed to get wet. Following the good recommendation of Sergey, and since the night was not cold, it was time for me to remove socks and trousers, saving them dry for later. It was my first experience of frogging with waders and underwear… Oh, well.

Fire-bellied toads were calling in big numbers, always a positive sign for finding treefrogs. After a long wait, surrounded by mosquitoes, the first treefrogs started to call all across a big flooded area, with some bushes and reeds. It was time to get into work. At the end, weather turned a bit colder than expected and frogs called for a short time. Anyway, we managed to catch 10 new frogs. I even caught two with my horrible, low-power, headlamp and leaking waders, pretty happy with that.

These frogs are really special for us. They will help us to compare them with the ones collected inside the Exclusion Zone, to use them as a reference of frogs living in areas never contaminated with radiation. Even more, a few of them will be used as the references for our genomic and transcriptomic studies. Really, really important, these clean frogs!!

This was also the last night that we had Sergey with us in the field for this time. He has to move back to the Exclusion Zone for more studies there, this time with birds. As always, it is a unique experience to share fieldwork with someone with the experience and knowledge of Sergey. Always and honor.

Our plan for today is to sample these frogs during the morning, and go to the field with Zenja. We will work again in totally clean areas, trying to increase our sampling size (and localities) of this kind of environment. This will be our last night of frog catching for the trip in Ukraine. So far, we have collected 100 frogs. How many we will catch today? What would be the grand total and the winner(s) of our game? Just a few hours to know!! Wish us luck!!

2018 Day 9. Moving day

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21 May 2018

Today it was a transition day for us here in the #ChernobylFrogs18 research project. After expending a full week inside the Chernobyl Exclusion Zone collecting treefrogs for our studies, it was time to move out and try catching frogs in areas never affected by the radioactive fallout after de 1986 accident in the Chernobyl Power Plant. So, we move to the east, to the city of Slavutych.

As usual, we started the day working in the lab obtaining samples from the 11 frogs collected the previous night. These frogs put our total for the trip, so far, in 90 frogs collected in six different localities inside the Exclusion Zone, two from high radiation area, one from medium, and three from low radiation levels. This is a great number, especially considering that the season this year is well advanced due to the intense warm in early spring, and that the weather was a bit colder than ideal. At least weather wasn’t like last year with temperatures barely over zero!!

Our job in the lab was very quick, the more days they pass the faster we go. We tried something new to communicate what we do here and used Twitter combining the hashtags #ChernobylFrogs18 and #liveScience to transmit live our work in the lab, resulting in 17 tweets. It was a fun experience, let’s see if it is possible to repeat something similar today from the field…

As usual, Sergey was in charge of the radiometer, measuring the radioactive content of our frogs, whereas Pablo, Jean-Marc and I, got the physiological and genetic samples from all the frogs.

We finished for lunch time, and quickly packed all our equipment again, from the tubes and scissors, to the cameras and the portable lamps. Everything needed for our work. This included also our dry shipper, refilled with more liquid nitrogen and very, very, full of samples. After waiting a while for our van, we moved to the main checkpoint of the Exclusion Zone. Our car was checked for radioactive contamination, our luggage was checked for radioactive contamination, and we were also checked for radioactive contamination. Our paperwork was in order, all permits stamped and approved. All clear, all fine. We passed the detection archs for radioactive contamination and out!!

Our drive from Chernobyl to Slavutych was a long, jumpy, drive. This drive usually takes 45 minutes by the direct road crossing Belarus, but due to bureaucratic issues it was impossible for us and our equipment and samples. We had to go all the way around to avoid crossing Belarus. At that means a 4 hour drive, first South to the outskirts of Kiev, and back North towards Slavutych. This was the bumpiest drive we have ever had, for kilometers and kilometers we were jumping like monkeys inside the van. It was even difficult to hear each other in the middle of the noise. Not the most confortable drive, really not. On Friday, we will do part of this drive again in our way to the airport… Anyway, on arrival it was Sergey waiting for us to show us our apartment for these days in Slavutych, and we finally drove through the city to a local restaurant in which we had an amazing dinner!!

Even better, after a full week of movement restrictions inside the Chernobyl Exclusion Zone, we were able to move freely through the city in our way back to the apartment!! Back to the free world!!

Our plan for today is going to explore areas near Slavutych and Chernihiv, looking for potential good places for frog sampling during the night. Weather forecast, again, is not super good, although it should improve during the day and the rest of our time here. It will be challenging trying to find a good place for treefrogs without much previous knowledge, but the area looks good and the search will be exciting, no doubt. Let’s see…

2018 Day 8. Easy frogs

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20 May 2018

An unusual night, very unusual night.. Weather forecast was bad, cold and probably rainy night, with no big hopes for catching more frogs in our last night inside Chernobyl Exclusion Zone. But… we ended up catching 11 new frogs in just twenty inmutes, in a locality visited last year, with plenty of time for photos, enjoy the landscape and taking it easy…

During the day, we worked pretty fast at the lab, finishing the last six frogs that remained from the previous day, and collecting all the samples from the seven individuals collected in clean area the previous night. Still, this take us the entire morning, with behavioral and coloration tests and the fourteen vials of different kind collected for each individual.

All blood analyses worked well, and we even had the possibility of running a few in duplicate to test the repetibility of the measures obtained with the iSTAT. Anyway, we finished just in time for lunch, once again prepared by Zenja, the best food (by far) we have had in Chernobyl. No more sampling to day, good food and a (finally) free afternoon ahead were the perfect excuse for opening the bottle of Alsatian white wine brought here by Jean-Marc. Excellent!!

We enjoyed a relaxed afternoon taking advantage of our WiFi connection at the hotel for keeping up with the family, the work outside here, and the world. Also for talking about our #ChernobylFrogs18 work in our Twitter accounts @pabloburraco, @bioecologie and @GOrizaola. Check there for more photos and comments!!

We also had time to properly store our data in our web repository, something essential when doing fieldwork, for a proper accountability of the work, and to avoid loosing data for unwanted reasons. So, we uploaded all the photos we have taken to our data during these days. Safety is an important part of Science, and not just the personal one.

After the usual early dinner, it was time for more fieldwork. Our last night inside Chernobyl Exclusion Zone, our last chance to add more frogs from the Zone for our study. Weather was supposed to be windy and cold with some possibilities of rain, not the best combination for catching frogs. Soon we realized that, although cold, the afternoon was not that bad, actually pretty nice.

We visited first a marsh area in the south side of the Exclusion Zone, a new locality for us, just to see that it was completely dry after the unusual warm weather of the spring. Anyway, light was perfect and we had plenty of time for taking photos, even Sergey took the camera out!!!

With that light and time before sunset, we had also the chance of taking a group photo (thanks again to Jean-Marc, our “official photographer “). Below, left to right, Jean-Marc, Zenja, me, Sergey and Pablo, the #ChernobylFrogs18 dream team!!

After this, and the removal of another tree from the middle of the road with a chainsaw, and the car stacked for a minute into the remains of trees, we arrived at a new location, the same location Sergey, Zenja, Pablo and I visited the first night last year. It was cold already, but as soon as we opened the car doors treefrog calls can be heard everywhere. No doubt that we were going to catch frogs, even more since we just needed a few. The idea was catching 8, but I quickly suggested 10 🙂 At the end, we managed to catch 11 new individuals in just twenty minutes, easy. Our total for the trip is now on 90, pretty, pretty good. Jean-Marc even took his camera to the field, so we have now these photos we never have, photos of fieldwork action. Pretty cool!!! In summary, a great, easy, really enjoyable night!!!

The plan for today is going quick to the lab, try to work as fast as possible sampling these 11 frogs, pack all our field and lab stuff, and try to be ready around lunchtime to leave the Exclusion Zone. Our goal now is to move outside the Zone, to Slavutych, a city created after the accident of the nuclear power plant to serve as home to people working in different jobs in the Zone. It’s a 45 minutes drive through Belarus (if no incidences at the border pass), but we can’t go that way due to the bureaucratic complexities of being three foreigners with a lot of weird scientific stuff (dry shipper, blood analyzer, samples, some chemicals…). Too complicated. So, it will take us four hours (!!!), driving back to Kiev and up again. Once in Slavutych, we will have three more nights for collecting frogs in areas never affected by radioactive contamination, essential to be used as references in many of our studies, particularly the genomic ones. If the weather is not too bad (cold forecasted) and we don’t arrive too late, we may try to go to the field tonight near Slavutych, if not we will have Tuesday and Wednesday ights for this. Let’s see how it goes in this new area!!

Working in clean areas only add the usual 3 microSv to our accumulated dose, for a total of 46 for the entire trip.

2018 Day 7. Frogs in the labyrinth

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19 May 2017

Today was a completely non-stop day. We spent 10 full hours working in the lab, sampling the frogs collected the previous night, just to jump into the car and go to the field for more. At the end, we collected seven more frogs from a completely new locality in the middle of the Exclusion Zone, a total labyrinth of trees and shrubs in which catching frogs was a bit of an odyssey.

We started the day still with the happy face of the 30 frogs collected the night before. This number, collected in two different localities (one new) is really important for having good sample size for some of the new traits that we are evaluating this year. Anyway, we knew that doing a full sampling of all these frogs will take a lot, a lot, of time…

As in previous days, we collected information and samples for about 30 different traits for each individual, from photos to evaluate coloration and swabs of the skin microbiome, to blood biochemical profile, and all kind of tissues for physiological and genomic analyses. A lot of work that takes more than 20 minutes per individual to Pablo, Jean-Marc and me. On top of that Sergey and Zenja are measuring individual dose rate with the radiometer. An amazingly complex and complete sampling!!

Even with all the quick work, we were unable to process the last six individuals before dinner time. Weather didn’t look too good during the day, big clouds, rain and a substantial drop in temperatures, far from the preferred conditions for catching treefrogs. We discussed if going to the field or not, and at the end we decided that we must go, to no surprise for our Ukrainian colleagues 🙂 This was probably the last night we had to catch more frogs inside the Exclusion Zone before moving out for more frog collection. So, there were no alternatives really.

We decided to move to an area around the center of the Exclusion Zone that we had not visited before. This area has really low levels of radioactive contamination, a good option for us after sampling already in three localities with medium and high levels. First places that we checked during the night look really good and with good activity of fire-bellied toads, but not a sign of treefrogs. After some more driving we arrived at a locality a bit in the middle of the forest, with treefrogs calling on the distance. There were not too many, but it was our best option.

Soon after entering into the pond it was clear that the place was difficult for catching frogs. A bog area full of shrubs and dead trees in which it was difficult to move due to all the branches closing the way. It was also a little nightmare of mosquitoes… Treefrogs were calling from the middle of the trees just to make things a bit more difficult. After a good while, we managed to catch seven frogs (5 Sergey!!, 2 me) at that was all. Not much, but this is a locality in which we had never worked before, so these samples are particularly important for our study. This is especially true for our population genetics studies, since this locality sits in the middle of the Exclusion Zone, covering a gap in our sampling map. Few, but important.

The night was cold and with no more frog activity, so we moved back to the cars and into Chernobyl city.

The plan for today is finish with the sampling of the six remaining from the previous night and fully process these seven individuals. The right amount of work to be busy but not that crazy busy as last day. On Monday, we will move out of the Exclusion Zone, towards Slavutych (the administrative center of the Radioecology Laboratory) for trying to catch frogs in never-contaminated areas. So, it’s not clear if we are going to be in the field during the night or not, since we wouldn’t have much time to process any frogs tomorrow morning. Weather forecast doesn’t look very good either. So, probably these frogs will be the last from the Zone for now, a total of 79 so far for the trip.

Being in clean areas means that we accumulate the standard 3 microSv today, for a total of 43 microSv for the trip. During the night we saw red deer, roe deer, white-breasted hedgehog and even a tortoise.

More tomorrow. Thanks for following our work in the Chernobyl Exclusion Zone, our attempt to tell scientific activities “live”.

2018 Day 6. Treefrog radioactive paradise

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18 May 2018

30. We collected 30 treefrogs on a single night!!

Once again the visit to the long ditch that runs parallel to the east side of Prypiat river on the nort if the Exclusion Zone give us the best results. This area, in which we have sampled in five different localities over the three years of our “Chernobyl frogs” project is definitely a paradise for amphibians, despite being an area with medium and high radiation levels.

We started our daily routines in the lab, sampling the thirteen frogs collected the previous night near Chernobyl city in an area of low radiation levels. Work on the lab was more or less as usual. There are never two equal days on a field lab, and there are always small incidences that need to be solved on the spot. Anyway, work on the lab was good, and there were not too many frogs to deal with.

Sergey even managed to pass all the frogs through the radiometer, so we have now good measurements of radioactive dose rate from all our frogs, including these from clean areas. Actually, it tourned out that not all these frogs were completely clean from radiation, and some showed low levels of contamination. Even more interesting.

Thanks to our acquired abilities (mostly, the amazing Jean-Marc’s technique of extracting blood!!) we were also able to run successful blood analyses for 12 out of the 13 frogs. These are amazing results that will tell us a lot of details about the physiological state of all these frogs. Results so far, after a quick view, are pretty consistent, showing variability on the diverse parameters we measure, but always between reasonable limits. We need to keep on adding more data here!!

By mid afternoon all the samples were collected. With the work, cryovials have started to accumulate in our dry shipper, and we even needed to add more liquid nitrogen there. This is one of the “luxuries ” of working here, with the International Radioecology Laboratory people, we have access (although not easy access) to good amounts of liquid nitrogen in the field, essential for storing all our samples for conducting physiological and genetic analyses.

Weather forecast was not too good for the night, with chance of rain, and temperatures going down. Anyway, after our early dinner we were ready to go to the field, with the idea of visiting different locations along the east bank of the Prypiat river, in the north of the Exclusion Zone. We have worked in this area since 2016, always with success. We started our journey to this area with good vibes, although the way was not as easy as expected, and we even need to take out a rather large tree from the middle of the path!!!

Once on location, we split our group in two teams, as in some previous days. We arrived to our first locality after a drive across the middle of nowhere, going around trees and up and down bumpy fields. This route, by the way, it would be impossible to do without the navigation skills of our Ukrainian colleagues that identified and marked this track for us many months ago. Thanks!!

After stopping, Evgeny (Žénja), Pablo and I moved to the first pond, and Sergey and Jean-Marc went back to sample on different localities. Our locality was located in rather high levels of radioactive contamination, with peaks around 20-30 microSv/hour (normal, background levels are around 0.1-0.3 microSv/hour). Soon after arriving we heard our first treefrog!! After twenty minutes more treefrogs were calling in the distance and we moved in. The area was easy for orientation, with few water channels going in circle. Water level was a bit higher than ideal, although enough (just barely enough) for working and crossing from side to side of the channels. The first frogs I caught were totally on the limit, captured with one hand while the other was putting the chest waders up in order to avoid water entry. Five centimeters deeper and they would have been out of reach…

Some frogs were loyal to their name and often calling from tree branches, a meter high from the water. Not always was easy to spot them there, and twice I had to use my left hand to catch them from a crisscross of branches (both times successfully!!). Through the phone we knew that our colleagues were not successful on their first locality, but managed to collect 16 frogs on a second place in which we have worked since 2016, on mid radiation levels.

At the end, we were also able to catch 14 frogs in our place, for an amazing total of 30 for the night, including a completely new locality. That’s fantastic!!! It was a particularly good night for Sergey and me, both with 11 frogs captured 🙂 These 30 frogs will definitely give us lot of things to do in the lab…

So, the plan for today is an intensive, super intensive, session of work in the lab with these frogs (let’s see if we can finish them before the night). If weather is good (forecast of rain for the entire day), we will try to capture more frogs in mid-high contamination areas close to Belarus. Anyway, long day ahead f work, sweet!!

Working on high contamination area resulted in a daily dose rate of 15 microSv, for a total of 40 for our trip. On the way today, we saw mousse, red deer, wild boar, white-tailed eagle…

2018 Day 5. Lucky thirteen

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17 May 2018

Frog activity is still rather low in the Zone, but last night we managed to catch 13 frogs in a completely new locality for us, our locality 13 for the project!! Activity in the lab run smoothly too. We only need now some good weather and a night of good frog activity to move forward.

The day started as usual with lab work, sampling the last frogs from our first high contamination locality that we didn’t had the time to finish the previous day, and the additional six collected in two localities the previous night. Our work in the lab is pretty well organized by now, and by lunchtime we were able to finish with all the frogs. We managed to run samples in the iSTAT blood analyzer for all the frogs but one. The technique by now is fully under control!! This will give us a really good indication of the physiological situation of our study frogs in the Zone.

We even had the time for taking some good photos of our frogs (wait for Jean-Marc great photos!!). These are the kind of photos we will keep on using again and again in our work, when given talks, publishing papers or having any communication activity. So, in a way, they are also important for the project!!

Talking about photos, Pablo continues with our colorimetric studies trying to assess the variation of frog coloration and if this changes with contamination level. So far, no many dark frogs this year, just a few.

Due to some bureaucratic limitations we didn’t had the chance of going too far during the night, so we had to work in low contamination areas during the night. Sergey drove us to a marsh area quite close to Chernobyl city we had not visited before.

We arrived to this place with plenty of time before dusk, just to listen to many water frogs and fire-bellied toads in the distance. No sign of our treefrogs, but we now that at this time we need to way until it gets dark for these frogs to start calling. So, we wait and wait. During this time, Sergey was able to imitate the call of a tawny owl (Strix aluco) good enough as to attract it at a very close range. That was fun.. At the same time, far away, we heard the first treefrog calling. Time to put the chest waders on and move through the reeds to find the place where they were calling.

Calling activity was low. Once there it was possible to listen to 4-5 males, good but not super good. At the end, after quite a lot of patience, the five of us managed to catch thirteen frogs in about an hour of moving back and forth through the pond. This is great. A completely new locality for the project, just in the middle of all our previous localities and with enough frogs as to do many good tests. Being a low contamination site is also quite good, since most of the frogs we had until now this season were from a highly contaminated place. Sweet!!

After this, we move to the same location in which we capture four frogs the previous night, but there was no additional activity there. Not even in another place in which we also collected frogs in 2016. The night started to be cold, so we decided to get back and try more luck in the next night.

So, the plan for today is lab work during the morning with these thirteen frogs, and try to go to some mid contaminated localities on the east bank of Prypiat river during the night. We have worked there the last two years and we know that this is good treefrog area. So, let’s see how it goes, and if the weather stays good (some rain forecasted and a bit colder than we wanted…).

Working in low contamination areas, we only accumulated 3 more microSv to our total dose, for a total of 25 microSv during the trip. During the night, we also saw Przewalski horses, a nice group on our last location, rumbling in the middle of the foggy night.

2018 Day 4 Busy day, poor night

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16 May 2018

Today was our first busy day at the lab, with 23 frogs to sample from the previous night, collected from a high contamination area. As always on the first day of a campaign it took us wuite some time to get everything organized, but finally it was a great (and exhausting lab day). The night was not so good… we visited three different localities but only managed to collect 6 more frogs.

During the day, it was time for us to sampling all the treefrogs. This time we were four people working together in the lab, which at the beginning is always something chaotic. Sergey was in charge of the radiometer, getting the radioactive dose rate for each individual. This is a process quite quick for highly contaminated animals, that will save us a lot of time (and money) on later analyses.

Jean-Marc was mostly in charge of the tricky extraction of blood for our blood physiology analyses with the iSTAT, and the sampling of frog saliva for hormonal analyses.

Pablo, same as last year, was responsible for the colorimetric evaluation, basic measurement of the frogs, and the obtention of samples for genomic analyses, telomeres length estimates, bone for skelotochronological estimation of the age of the frogs, and (new this year) some behavioral tests.

Finally, I took samples for analyzing skin and gut microbiome, oxidative stress, DNA damage, sperm and charging the cartridges for running the iSTAT blood analyzer.

At the beginning this was way too complicated to organize. Too many people doing too many things with the same frog. But, after a few frogs things started to run, although a bit slowly, quite ok for the amount of samples per individual (more than 30 different parameters!!!). Working with the iSTAT was a nightmare at the beginning. Cartridge after cartridge went to the bin because of lack of enough blood, bubbles when we tried to fill the cartridge and more. At the cost of 13€ per cartridge… Then, Jean-Marc modified a bit the blood extraction technique, getting more blood, I modified a bit the insertion technique… and with our frog 12-6 (locality 12, frog number 6), everything worked!! The song of the iSTAT printer when a sample is successful is now by far our favorite song in the house!! Finally, we managed to get results from 7 out of the 12 last frogs, after six consecutive failures!!

At the end, they were too many frogs and too little time. So, it was not possible for us to sample the last five frogs. We left them for the next morning. It was time for us to go again to the field, looking for more frogs in areas of low/medium contamination. We divided us in two teams, so we can check more localities at once. Anyway, the locality that Pablo and I visited with Evgeny, was full of water frogs, but nothing more. Not a single treefrog, not even fire-bellied toads… The area, located in the center of the Exclusion Zone, was part of a former youth camp during the soviet times, all abandoned now. Fate for Sergey and Jean-Marc was pretty similar, two treefrogs in a medium contamination place.

We decided to move for a clean locality that we visited during our 2016 trip, with the idea of collecting frogs there to complete some of the different parameters that we have added on the last years. But neither there we had much success. We were able to locate and capture four male treefrogs, in the middle of big noisy calls from water frogs and fire-bellied toads. It was midnight and no more frogs were calling, so we decided to go back and hope for better luck on next nights. So, a total of six new frogs to add for the trip, for a total of 29.

The plan for today is to finish sampling the last frogs of the first locality and these six collected last night. Hopefully the iSTAT will keep on working well and can complete all the sampling. During the night we plan to go to some localities near Chernobyl city and other areas in low/no contamination. Weather forecast looks semi-OK, warm but with chances of rain in the afternoon. So, let’s see if frogs are active, and how many!!

Working in areas with no contamination means that we only accumulated 3 microSv extra, for a total of 22 for the trip. During the night we managed to see one Przewalski horse, red dear, red fox and many hares.

Here some extra photos of the lab work, our radiometer working with a frog, and one of our Eastern treefrogs (Hyla orientalis).