New Publication – IV oxytocin causes pro-social behaviour in seals

Grey seals on the Isle of May, Scotland. Staying together is important for mother-infant pairs, especially on a dangerous seal colony.

Link to article: http://rspb.royalsocietypublishing.org/content/284/1855/20170554

Or read the summary here on this site.

This week has turned out to be a hectic but good one, I’ve returned from the University of Liege just in time for my next paper to be published in the Proceedings of the Royal Society B. The paper comes from the research in my NERC funded PhD with the Sea Mammal Research Unit, University of St Andrews on the hormone oxytocin and its impacts on social and maternal behaviour, rather than the pollutant research I’m currently doing with the PHATS team. Like much of my work, the study was done with weaned grey seal pups on the Isle of May, and involved giving the seals specially designed doses of oxytocin to see what (if any) social behaviours the hormone affected.

There have been lots of studies that show certain behaviours are linked to oxytocin concentrations (including some of my own grey seal work!), but the problem with correlations is that you have no idea which side of the relationship is driving things. For example, it would be impossible to tell using only correlations whether increased social behaviours are causing high oxytocin levels, or high oxytocin levels are triggering more social behaviours. Understanding causality in such hormone-behaviour relationships is important so you can identify the ‘cause’ and the ‘effect’ within the correlation. It can be difficult to do outside of laboratory settings however, as the only way to test for causality is to either increase the hormone’s concentration in an individual via manipulations or knock out the functionality of the hormone using antagonists. Due to these difficulties, there is only one study (apart from the one I published today) that has ever given oxytocin to wild individuals, and while they did find changes in pro-social behaviours they had no prior knowledge of the natural oxytocin-behaviour systems in their study animals.

We know high oxytocin grey seal mothers stay closer to their pups, but does the hormone cause the behaviour or does being near to their pup for more time cause greater oxytocin release?

In our study we were testing whether oxytocin triggers individuals to stay close to each other, as we know from grey seal mothers that the higher their oxytocin concentrations, the more time they spend close to their pups. We gave oxytocin and saline (control) treatments to weaned grey seals that had never previously met, and recorded their behaviours after the treatments. We found that oxytocin not only triggered individuals that had never met before to spend more time together, but also reduced aggression between the two and the amount the seals investigated each other, an indication of familiarity. This makes our study the first to verify a naturally existing oxytocin-behaviour relationship in wild individuals, which is very exciting. Studies like this have been done in captivity using domestic, laboratory or zoo animals but it’s crucial to study behaviour and physiology in natural settings with wild individuals, as no matter how hard you try you will never truly re-create all the complex aspects of wild environments in a captive setting.

Weaned grey seal pups associating on the Isle of May, Scotland

The treatments were all given intravenously (IV), as the more common, intranasal route of oxytocin manipulation was not possible with the weaned seal pups; they can close their nostrils and hold their breath for a long time! The success of this route of administering the manipulation means that other animal species, that may not be suitable for intranasal manipulations, could potentially have similar studies done on them in the future to help us understand more about oxytocin’s important role in bonding and behaviour. We also spent considerable effort designing the oxytocin dose given to the seals to be as low as possible while still having a behavioural affect. Many doses used in the scientific literature are much higher than natural concentrations, and there are concerns that generating high levels in study individuals could trigger behaviours that would never naturally happen, or have unexpected, and unwanted, side effects.

Weaned grey seal pups having a disagreement on the Isle of May. Reduction of aggression between familiar individuals happens naturally without oxytocin release in seals, but manipulations also trigger this behavioural change with seals that are complete strangers.

Despite the effort we went to in replicating natural oxytocin concentrations as much as possible for our study, the treatments still triggered some behaviours that are not naturally correlated to oxytocin release in seals. Low aggression and reduction of investigative behaviours are indications that seals recognise each other, and naturally occur after several days of living together, independently of oxytocin release. The behavioural changes in our study seals also unexpectedly persisted for several days, long after the dose would have been metabolised and broken down in the bloodstream. These unexpected effects show that we still have a lot to learn about oxytocin’s role in the formation and maintenance of social and parental bonds. If the hormone is going to be used to safely and successfully treat human psychological conditions such as schizophrenia, autism spectrum disorders and post traumatic stress disorder (and there have already been clinical oxytocin trials for such conditions in human subjects), then more research is needed into the powerful effects oxytocin can have on our behaviour and neurobiology.

Weaned grey seal pup on the Isle of May.

Liege 2017 – A brief introduction to blubber tissue

Minced blubber biopsies in cells ready to be capped and put through accelerated solvent extraction, to obtain all the lipids from a samples for further analysis.
Weighing a ASE vial after ASE is finished and the solvent has been evapourated off to calculate lipid mass in the sample. This is from one of Foxtrot’s biopsies from last year, and it had 0.42g of lipid in it, meaning 80% of the original biopsy was fat.

Well my first week at the University of Liege working with CART has flown by, and I’ve been working on the blubber biopsies we collected from the grey seals last year on the Isle of May. All the lipids (fats) need to be extracted from the blubber tissue before we can move forward with the pollutant analysis, so all the samples must be carefully prepared and put through Accelerated Solvent Extraction (ASE). This process uses high pressure and temperature conditions plus chemicals called solvents (like hexane and acetone) to remove all the lipids from the sample in the cells. This process gives us a completely liquid solution of lipids and solvents at the end of it, and we can then evaporate the solvent to leave just the lipids from our sample. This step is important as it gives the lipid mass of our sample, and allows us to work out how many nanograms of pollutant per gram of lipid in our sample there is (ng/g) . While ASE of our samples is an important step in the lab work, there isn’t really much more to say about it so I’m going to use the rest of the blog this week to give a brief introduction to blubber tissue, a crucial part of the anatomy of all marine mammal species worldwide.

Blubber tissue enables marine mammals to endure cold aquatic environments, like this Orca family living off the coast of Iceland.

All marine mammals, from the largest whale to the smallest seal, have a layer of fat underneath their skin called blubber. This layer of fat is extremely important for the survival of marine mammals for two reasons:

  1. It enables them to keep warm (thermoregulate) in freezing oceans.
  2. It provides a store of energy for individuals to utilise when they are not feeding, which happens in many marine mammal species at various points throughout their lives due to breeding or moulting.
A blubber biopsy from one of our seals. Blubber samples for pollutant analysis must be stored in glass or wrapped in foil to prevent loss of pollutants from the sample to any plastic they come into contact with.

Fat tissue deposits in all animal species perform these same two functions, however other species frequently have additional ways to thermoregulate (such as fur in land mammals) or do not endure long periods of fasting repeatedly while migrating or breeding as many marine mammals do. The importance of this tissue has lead to substantial blubber thickness evolving in marine mammals, and a stratified structure throughout the depth of the tissue is present so that it can perform both functions at the same time. Typically, blubber tissue can be roughly divided into three sections as you go from the part closest to the skin (the outer blubber) to the part closest to the inside of the seal (the inner blubber). The inner blubber is the most metabolically active, and this is where lipids are mobilised to provide energy for an individual when it either cannot find food or is purposefully fasting. The mid blubber is the most variable in thickness across individual marine mammals, and in thin individuals can be completely absent. It is thought it acts as a more long term storage tissue, and that its thickness is influenced by seasonal food availability. The outer blubber is typically of stable thickness within a species regardless of the nutritional state of an individual, and is thought to be primarily for thermoregulation. Hence even starving individuals will always have some blubber tissue to keep them from freezing, as the outer blubber is not mobilised as an energetic resource.

Blubber is a fascinating tissue to study and several different approaches can be used to analyse it in many contexts, like this recent study by one of my friends, Joanna Kershaw, who measured the hormone cortisol in blubber from porpoises to validate it’s use as a biomarker of body condition. The PHATS project I work on uses both established techniques (investigating pollutant concentrations) and novel protocols (the explant approach for tissue culture experiments that our team leader pioneered in seals) to make the most of the blubber samples we collect from our study animals to explore the prevalence of persistent organic pollutants in the marine environment and it’s impact on energy balance in seals.

A particulary tubby looking Charlie from last year’s study group on the Isle of May. Her fat reserves in her blubber layer will help her survive the tough first year at sea she faces when leaving the breeding colony.

MEANWHILE I am settling back into Liege life quite happily outside of the lab. I am not staying in the university accomodation this year, and have a lovely little flat not far from the campus to retreat to. In my time away from the lab I’m trying to keep up with the usual paper and grant writting that all resarchers need to keep on top of, plus greatly enjoying bebing reunited with the amazing macaroons they make here! Seriously, why can’t they be this good in Scotland…

Happiness is macaroons =D

PHATS project progress, ELISA validations and my return to Liege

Moulting grey seals hauled out on the Isle of May
Remember please send all ringed shag sightings to shags@ceh.ac.uk

Welcome back to my corner of the internet and the PHATS blog! The first four months of 2017 have flown by as the team has headed back into their labs to analyse all the samples we collected over the winter on the Isle of May grey seal breeding colony (to read about our fieldwork adventures, see these blogs here). I was lucky enough to escape to perform a survey of the Isle of May in January, to see if there were any grey seals that were moulting early in the year. There were plenty of them as it turned out, which bodes well for the fieldwork we are planning next year to try and look at moulting seal physiology. The island already looks so different to how it was when the seals breed there in the winter, much greener and all the seabirds are starting to come back. The cliffs were lined with guillemots, razorbills and shags; some were even getting started on gathering nesting material. The puffins had not returned yet, they arrive later in the year closer to summer, but we did see a few on the water during the boat crossing to the island.

Large grey seal haul out on the North East coast of the Isle of May
Is there anything better than well organised samples?

Inside the lab, I’ve been working on biochemical analysis of the cell culture media from all the blubber sample experiments last year (see here for more info) and am now two thirds through the samples we generated. By measuring the metabolic profiles of the various blubber culture experiments, we can see if the pollutant or hormone treatments had any impact on the blubber cells we collected from the seals. I’m also working on validating ELISAs (Enzyme-linked immunosorbent assays) to detect a variety of hormones in the blood samples we collected from the seals last year, so we can see if an individual’s hormone profiles are linked to their pollutant burden.

Using ELISAs on wild animal species like grey seals can be tricky, as they use antibodies as part of a binding process to detect the hormone you are interested in. These antibodies will have come from a specific species of mammal, usually rodent or domestic animal species that lots of scientists study, and the company making the ELISA kit will provide a list of species they know the kit works with. As a hormone’s protein structure is not always the same in different species of animals, the antibodies used in a widely available ELISA kit may not react properly with samples from unusual species that has never been tested with that kit before. Unfortunately seals often fall into the ‘unusual’ category, so we need to test the kits (validate them) before we use them to run lots of our samples, to make sure the results we are getting from the kits are accurate. There are several things to check when validating an ELISA, some of the most important are:

  1. Test for linearity. By diluting some of your samples (e.g. to half the concentration, then a quarter, then an eighth etc) you can make a serial dilution series to run on the kit. You can then see whether the curve the dilution series produces is parallel to the standard curve, which is what the kit uses to determine hormone concentrations. If the curve is not parallel to the standard curve, then the hormone in your samples is not binding correctly to the kit components.
  2. Test for recovery. By spiking a sample with a known quantity of the hormone you are interested in studying, you can tell how much the kit is detecting and how much is ‘lost’ during the analysis process.
  3. Test for consistency across kits and within kits. Many studies have lots of samples and need to use more than one kit to analyse them all. You must make sure the results of one kit are comparable to the others (inter-assay coefficient of variance), and the easiest way to do this is to run the same sample on each kit you use. You can then calculate the coefficient of variance across all the kits you have run. It’s also important to check the kit’s internal consistency (intra-assay coefficient of variance) by running one sample multiple times on a plate and seeing how similar the results are, and by running all samples in duplicate on the kit.

Our seal blood samples are proving to be rather tricky currently, and we’re still working on which are the best kits to use to measure the hormones we are interested in. The biochemistry analysis is going very well though, and we’re all looking forward to having some data to play with in the coming months.

Two ELISA plates at different stages of incubation. The different intensities of colour indicate different concentrations of hormone.

The biochemistry and ELISA work I’m doing is currently on hold however, as I have returned to the University of Liege in Belgium to work with the Centre de Recherche Analytique et Technologique (CART) to detect the amounts of persistent organic pollutants (POPs) in the blubber of our study seals from the Isle of May last year. We will be using Gas Chromatography – Mass Spectrometry (GC-MS) again, which requires a lengthy extraction and clean up process before the blubber tissue can be analysed (see these blogs from last year for more details) so I will be here for a month to work on our samples (and I will update the blog every week while I am visiting CART). It is always interesting, and more than a little sad, to find out how many pollutants all the seals have inside them after we got to know so well during the field season…

Unfortunately Alpha and Kilo, like all young marine mammals, will have large concentrations of POPs in their tissue from the high fat milk they drink from their mothers.

Isle of May 2016 – Goodbye to the Island!

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A curious weaner watches us load up the boat and leave the island

The time has come for the PHATS team to pack up and go home as almost all the grey seals have left the Isle of May and returned to sea. We have all got back to mainland safe and sound, and everyone is looking forward to some well earned rest after almost two months of fieldwork. All of our laboratory equipment and samples were packed up and shipped off the island without incident, and everything is now at our labs at Abertay University and the Sea Mammal Research Unit, ready for analysis in the new year.

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Walking down to meet the boat at sunrise, ready to leave the island yesterday morning

There were only nine of our study weaners still on the island during our last survey, and the majority of them are on the edges of the colony ready to go to sea. Oscar is still hanging out in his pool off the south end of the island, and Papa and X-ray have both made it to the bottom of the southern cliffs and are playing and dozing with a bunch of other weaners there. Hopefully they will all find their way out to sea and learn to forage and catch fish in the coming months. Even if they do survive their risky first year, it’s sadly unlikely that we’ll see them anytime soon on the island again. Grey seals usually don’t come back to breed for at least five years if they are female and even longer if they are male. If they do return to the colony however, we’ll be able to identify them from the number on their orange flipper tags, and if you see any seals with tags please do let us know so we can work out which of our study animals you saw.

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Papa (right foreground) with a gang of other weaners, ready to go to sea at the base of the south cliffs on the Isle of May

These kind of mark-recapture methods are used in all sorts of studies on many different animal species, which I was reminded of during the last weaner survey before we left. While searching the southern cliffs I came across a shag with a ring on its leg, which marks it as one of the study individuals the Centre for Ecology and Hydrology has captured, ringed and released back into the wild to help collect data on seabird population dynamics and behaviour. If you see a ringed shag (the leg rings can be several different colours, not just yellow with black writing like the one I saw below) please email the sighting information to shags@ceh.ac.uk and if you are interested in the project’s work then you can keep track of what is happening through their twitter feed @CEHseabirds. This particular bird was ringed as a chick in 2000 on the Isle of May, which makes her 16 years old and she has raised 26 chicks so far in her life (information courtesy of the Centre for Ecology and Hydrology).

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The shag I saw on the southern cliffs of the Isle of May, with ‘DDJ’ on her coloured leg band

The PHATS team will be back out in the field in January 2017, when we will return to the Isle of May to look for moulting grey seals. The majority of individuals will enter the moult later in the year at around March/April time, but there are always some seals that start early. We will resume blog updates then, so in the meantime we all wish you Merry Christmas, a Happy New Year and hopefully see you in 2017!

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Goodbye for now… grey seals hauled out on the north eastern shore of the island, with the Low Light, the Beacon and the Main Light above them.

Isle of May 2016 – A lover or a fighter? Male mating tactics and the end of the field season is nigh…

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Male grey seals fighting for access to females on the Isle of May

Thus far in this blog I’ve mostly focused on talking about the grey seal mothers and pups which live on the breeding colony here on the Isle of May. The PHATS project only involves collecting data from mothers and pups at various ages, hence I’ve somewhat ignored the other occupants of the colony up to now. However, the other individuals on the colony show plenty of interesting behvaiours too, so this particular blog is going to be about the male grey seals that live on the Isle of May at this time of year, waiting to try to compete for opertunities to mate with the females.

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A male grey seal (right) with two females with pups on the Isle of May. Male grey seals are not significantly larger than the females like in some other pinniped species.

Male mating tactics in species of pinnipeds (seals and sea lions) tend to depend on how sexually dimorphic (differences in size or external features like horns between males and females) the species is and the type of habitat they breed in. Some pinniped species, such as elephant seals, have males that are much larger than the females of their species. This facilitates establishing a territory on a breeding colony that contains as many females within it as possible, defending it from rival males and keeping lots of females inside. These huge males will fight any other males that intrude in the territory to prevent them from entering and mating with the females within. Other pinniped species that breed on ice in the north and south poles of the world use physical features like breathing holes to define where males establish territories, like Weddell seals in Antarctica. Male Weddell seals are a similar size to their female counterparts and use complex vocal displays underwater to defend their territories around breathing holes rather than physically fighting with other males.

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Some pinniped species are highly sexually dimorphic in terms of body size, this is a male New Zeland fur seal which can be three to five times larger than the females.

Grey seal males show a variety of mating strategies on and around breeding colonies. Males in this species are not much bigger than the females, so creating and guarding large groups of females to mate with (a harem) is not possible. Some males do establish loosely defined territories across various areas of the breeding colonies, but these do not seem to be anchored to physcial features or resources in the colony. The males that compete to stay within the colony amongst the females are called as tenured males and they typically the largest males on the colony, fasting to hold their position in the colony for as long as they physically can before they must return to sea to feed. However, there are alternative mating strategies that males can use to gain opportunities to access receptive females, including intercepting females that are returning to sea when leaving the breeding colony and mating with females at sea. Some male grey seals do return to sea and engage in  shallow and deep diving behaviour during the breeding season, but whether these can be attributed to feeding, displaying to attract mates or aquatic mating with females is not currently know. Therefore there seems to be a variety of mating strategies that individual male grey seals can employ on breeding colonies to gain access to females, and future research will hopefully help us understand which are the most successful and how selection pressures have encouraged the development of the different mating tactics.

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Being big and winning fights is not the only way for grey seals males to win access to breeding females, but it’s frequently the most conspicuous!
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Delta playing in a tidal pool, the next time the tide comes in he will go out to sea with the receding tide.

The PHATS research project is almost complete as the end of the field season is quickly approaching. We only have two samples left to collect, then we will pack up and head off the island just in time for Christmas! Over half our study weaners have now gone to sea, there are only 13 left on the island. The oldest remaining seal is India at 49 days old, but we still have some fairly young weaners like Eel (not in the phonetic alphabet I know but we always run out of alphabet and have to start giving study weaners name codes like AA, BB, CC… and Eel is EE!) who is only 26 days old and still over 50kg. She could hang around the island for quite a long time with all that blubber mass, we have known weaners to remain on the colony for over a month after weaning.

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Eel checking me out. She is massive at over 50kg!
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Oscar in a tidal pool, meeting other weaners and learning how to swim before the tide comes in and he goes to sea.

Sadly, while doing the daily colony survey to find our study weaners I also found a weaner that had plastic wrapped around his neck. I managed to deprive him of his  ‘necklace’ and he swam off into a pool without serious harm, but it’s a sad reminder of how abundant marine litter is, and how it can cause deadly problems for marine life around our shores. Please be aware of your litter, whether you are on the coast or anywhere outdoors as many speices of creatures can end up trapped or eating the things we throw away, causing severe health problems that they frequently cannot alleviate by themselves.

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Marine litter causes significant health problems, and death, for many kinds of marine life from birds, to turtles to marine mammals. Please pick up your trash and dispose of it properly when you are by the sea!

MEANWHILE Away from the colony,we had some visitors for the day this week as some people from Scottish Natural Heritage came out to the island to prepare for the field station closing down for the winter. There are people living on the island almost every month of the year as bird researchers come out in the spring and stay studying the birds until early october, then the seal researchers move in until mid December. However, between December and spring there are a few weeks when no one is living and working at the field station so everything had to be cleaned, packed away and locked up against the winter storms. It was nice to see some new faces after 2 months of seclusion and everyone had a great day despite the choppy sea conditions getting to and from the island!

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The SNH guys leaving the island after their visit, hopefully the trip back was drier than the trip out!

The island has also become noticeably more noisy in the last week as the cliffs have become covered in sea birds returning from offshore early to roost. We currently have lots of guillemots

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Noisy fulmars on the Isle of May cliffs

and fulmars on the cliffs and it is lovely to see some more life on the island, especially now there are so few seals around. With the seals departing, and our project coming to an end this year, we will likely be leaving the island in the next week to head home. Going back to mainland after a few months away from society is always a little strange, but everyone is looking forward to seeing their loved ones (and getting regular showers again!).

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Fulmar in flight around the cliffs, the strong winds around the island provide excellent opertunities to watch the fulmars soaring around, looking for roosting sites.

Isle of May 2016 – Late season births and Charlie goes to sea!

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Grey seal mother with her newborn pup on the Isle of May, taken this week.

Even though it’s december and the vast majority of the seals on the Isle of May breeding colony have already raised their pups and returned to the sea, we are still seeing a few newborn pups from mothers that are late season breeders. Most of the colony is quiet now as the majority of adult females have gone, leaving their weaned pups (weaners) in their 2-3 week post-weaning fast (see this blog for more details about the post-weaning fast in seal pups).

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Things are all quiet now it’s late season, just lots and lots of weaners playing and dozing everywhere!

We don’t really know why some females give birth so late in the season, but doing so tends to have a big impact on how likely they are to raise their pups to weaning, with late season mothers typically having lower pupping success than early season mothers. While there are very few adult females on the colony, and therefore less problems with aggressive neighboring females, there are still plenty of adult males around looking to mate. The ratio of females to males on the colony changes through out the breeding season, and goes from lots of females to every male in the early season to at least one male per female late in the season, if not more. This means that a female trying to raise a pup in the late season has to deal with a lot more male harassment than the females in the early part of the season, and as a result she spends less time nursing and caring for her pup. There is also evidence that females breeding in the late part of the season are younger and therefore are more inexperienced mothers, but what age females start to transition to become early season breeders, if they do at all, is still a mystery, especially as most of our study mothers seem to give birth exactly the same time each year, to the day in some cases.

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Male harassment can be a big problem for grey seal mothers trying to raise a pup

Back at the PHATS lab, things are starting to wind down. We had to ship all the samples we had generated so far from the cell culture experiments off the island this week as we had filled the minus 80 freezer on the island! The tissue, media and blood samples we’ve collected over the last 6 weeks will give us plenty to study back on the mainland in the new year. Our daily surveys of the study weaners have shown that two of our weaners, Charlie and Alpha, have left the island and gone to sea! Charlie was 35 days old when she left the colony, and Alpha was 43 days old. Some of our study weaners don’t seem very keen to go, both Bravo and Delta are 43 days old and still happily playing around on the island! Some of our seals are making their way through the colony and the rocky tidal region of the shore towards sea, like Foxtrot who is 40 days old and is on the southern edge of the island.

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Delta playing around in a pool
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Foxtrot in amongst the rocks of the southern edge of the island
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Bye everyone!

MEANWHILE the research team on the island has dropped to just 4 people as half the team left the island this week to return home, the remaining two researchers from the Durham University seal behaviour team and two research assistants from the Sea Mammal Research Unit of St Andrews. We also were lucky enought to be visited by one of the bird researchers from the Center for Ecology and Hydrology again, as they keep track of the ringed sea birds (shags) on the island during the winter. During this visit they caught and ringed a wild short-eared owl and were kind enought to show us before they released her, beautiful!

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Short eared owl that was ringed on the Isle of May, just before release

 

Isle of May 2016 – Go to sea or stay and play? Weaner dilemmas

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PHATS team photo! From left to right, Holly Armstrong (research assistant), Dr Kimberley Bennett (team leader) and myself (research fellow) all geared up for an afternoon of studying seals on the Isle of May.

It’s currently my favourite time of year on grey seal breeding colonies, when the  weaned pups (weaners) start to out number the adults on the island. I do enjoy watching the huge variety of behaviour the adults display, from the caring maternal behaviour mothers exhibit to the impressive battles males fight for access to females, but the weaners are without a doubt my favourite. This is because they typically spend several weeks on the island after weaning in a post-weaning fast, where they stay on the breeding colony despite there being no food for them, and they usually spend this time playing and exploring their surroundings. They can be very curious, and some will even come right up to you when you’re hiding in the colony to see what you are, play with youre belongings or even fall asleep on you!

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While hiding amongst the stone walled parts of the Isle of May to survey the seals, you can get unexpected visitors at this time of year!

Post weaning fasts are common in Phocid (true seal) seal species around the world, from the huge elephant seals to the two species we get in the UK, grey and harbour seals. No one really knows conclusively why weaners stay on the breeding colony for several weeks after their mothers wean them, although there are lots of theories. Grey seal pups can lose half a kilogram of mass every day they remain on the colony after weaning, and as having large amounts of blubber increases the chances that the pup will survive it’s first year of life, it seems very strange that pups stay on the island rather than go to sea to learn to forage and catch fish. However, there is evidence that during the post weaning fast, pups undergo physiological changes which prepare them for going to sea for the first time, so they can hold their breath and perform deep dives so they can catch fish successfully. In grey seal pups, the amount of oxygen they can store in their blood increases during the post weaning fast due to changes in the volume of blood they have and its components, with increases in substances such as hemoglobin that bind oxygen. The more oxygen a pup can store in it’s blood before it dives, the longer it can hold it’s breath and forage underwater, and in theory the more successful at catching fish it will be.

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Going to sea and learning to survive there is a crucial step in a weaner’s life, but one with many difficulties and dangers.

There are still many unanswered questions about the post weaning fast phocid seal pups go through and what happens once weaners leave the colony for the first time. To try and answer some of these questions, one of the senior scientists (Dr Sean Twiss of Durham University) here on the Isle of May is currently looking for a PhD student to investigate “Patterns of individual variation in post-weaning behvaiour in wild grey seals“, and the project is being co-supervised by my boss, PHATS team leader Dr Kimberley Bennett of Abertay University! The deadline for applicants is the 20th January 2017, and if you are interested, the details of the project and how to apply are found here. Whoever gets the PhD will get to come out to the Isle of May next year and will meet me and the rest of the PHATS team! Matt Carter of Plymouth University, another PhD student that Dr Bennett co-supervises, is also currently investigating the development of foraging behaviour in pups leaving breeding colonies, so hopefully in the coming years we will gain a much better understanding of the factors driving weaner behaviour before and after they go to sea for the first time.

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Charlie is getting leaner as she loses blubber during her post weaning fast, which has been 17 days long so far, and at 35 days old she may go to sea soon and leave the island.
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Our CO2 incubator filled with plates of blubber tissue explants for our pollutant work.

In the lab here on the island the PHATS project is just over halfway to completion this year. As it gets later in the breeding season we are getting more and more study weaners to keep an eye on, although some of the first weaners that we included in our study are approaching the time when they may go to sea for the first time, at about a month of age. Charlie is now 35 days old and much leaner than when she first weaned from her mother 17 days ago, having lost 9.6kg across those 17 days. This seems like a lot, but considering she put on 22.2kg of blubber in the 18 days she was with her mother, she is still large enough to successfully go to sea. Kilo has now fully moulted and has a very distinct pattern of spots that we can identify him from. We also have a new study weaner (Oscar) that has moulted and turned out to be completely black!

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Oscar, one of our latest study weaners.
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Kilo, now fully moulted with spotty fur, playing with the plant life on the island

MEANWHILE away from the colony, it briefly got a little bit harder to escape from the seals as a female came right up outside our doorstep with her pup for the first time! She has thankfully moved on a little further from the house so we don’t disturb her, but it was a suprise to open the door one morning and find the pair of them staring at us!

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Suprise! A grey seal mother has never had a pup right by the house before on the Isle of May

Sadly two members of the research team here have had to leave the island in the last week, Dr Twiss and Dr Bennett both had to return to the mainland to resume their teaching duties at their universities. They will be missed, and it marks the beginning of the end of the field season as the majority of the researchers here will also leave in the coming week. The PHATS team will stick it out until mid December, but it’s always sad to see people go!

Finally if you’d like to see Dr Bennett talking about seals or the PHATS project, she will be giving some public presentations in the coming weeks in Scotland:

6pm, Tuesday the 13th December at the D’Arcy Thompson Zoology museum at Dundee University: Public talk on seal biology

10.00-16.30, Wednesday the 14th December in the Ninth Scottish Symposium on Environmental Analytical Chemistry at the Dalhousie Building, Dundee University: presentation of persistent organic pollutant findings from the PHATS project

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Dr Bennett and Dr Twiss setting off from Altarstanes to return home.

Isle of May 2016 – Pup survival 101: get fat fast, lose your fluff, find some fish

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PHATS study pups Foxtrot (in the background) and Golf (in the foreground), both at different stages of moulting their white baby fur.

As it gets later into the breeding season here on the Isle of May, more and more of the grey seal mothers are weaning their pups and returning to sea to fish. Grey seals, like most species of true seals (called ‘phocids‘, including all earless seals), have one of the shortest dependant periods (the time the infant has to stay with the mother to survive) of all mammals, just 18 days. After that, mothers leave the colony and it’s up to the pups to look after themselves. Left to their own devices, newly weaned pups (called ‘weaners‘) soon find their way out of the colony or into nooks and crannies out of the way of the adult seals where they can rest and play, moulting their white baby fur (called lanugo) so they are ready to go to sea for the first time.

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A fully moulted grey seal ‘weaner’ on the Donna Nook breeding colony in Lincolnshire.

Moulting pups are usually much larger than when they were born despite only being 18 days old. Grey seal mothers produce milk with such a high fat content that pups can put on over 1.5kg a day in blubber tissue. Pups that are born at just over 10kg can therefore easily reach over 40kg in mass at weaning, and for seal pups the fatter they are at weaning the better as it means they are more likely to survive their first year of life. During their first year, a weaner’s body composition will dramatically change and they will go from the fat, round shape like in the above photo to a leaner look, like the yearling in the picture below. As a juvenile, a grey seal’s body composition is only about 12% fat of its total mass, a huge difference to the 45% fat individuals can have as weaners. These big changes in body composition are completely natural for the seals, but it means that it is vital for their fat tissue to function correctly both when they are rapidly generating blubber or mobilising it when they don’t have any food. That’s why the PHATS team are investigating whether the high persistent organic pollutant (POPs) burden that all marine mammals have in their blubber tissue is disrupting the seal’s ability to generate and utilise their fat reserves.

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A lean yearling grey seal on the coast of North Rona, Scotland

We now have 12 study weaners this year, all named after the phonetic alphabet as their identifying code in our project is a single letter. The largest of our pups is Lima, a male weighing 54kg! I introduced you to Charlie in the last blog when she had just weaned from her mother and still had lots of lanugo covering her body, she’s changed a lot in the last week! She’s moulted completely now and has really pale fur with subtle spots.

Charlie now she has fully moulted, these spotty patterns will stay the same on her fur for her entire life.

The other weaners in our study vary from pups that are almost completely covered in lanugo to fully moulted, and its interesting seeing the different patterns the weaners have hiding under their white baby fur. Here are some more introductions to our new study weaners, you can see Foxtrot (female) and Golf (male) in the photo at the top of the blog, and below are pictures of Alpha (male), the oldest pup in our study, and Kilo (male) snuggled up together and Delta (male) playing around with his flippers.

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Alpha (left) and Kilo (right) interacting
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Delta playing around
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The super moon the night after it was at its largest

MEANWHILE away from the seals, we were all very keen to see the super moon, unfortunately the weather on the island did not co-operate and we didn’t get to see it in it’s full glory. We did all manage to see it before the sun rose the following morning and the next night, which was still very impressive!

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The high tides from the super moon submerging the pier at Kirkhaven harbour on the island.

Even though we didn’t get to see the super moon properly, we still felt it’s impact as we had huge tides on the island, and all the seals near the coast found the ocean encroaching where it doesn’t normally reach! The resulted in lots of seal pups getting swimming lessons with worried mothers in tow, and even some underwater nursing attempts.

Sadly it’s getting to the time in the season where some of the team have to think about leaving the island, including our team leader Dr Kimberley Bennett who has to go back to the Abertay University to continue teaching. It’s likely Holly and I will stay on the island until mid December to complete our work on the weaners, so we’ll be getting to know them very well in the coming weeks!

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A grey seal mother trying to feed her pup during the high tides generated by the super moon

Isle of May 2016 – Beginners guide to pups and meet Charlie!

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A healthy pup over a week old playing with it’s flippers on the Isle of May

It’s now a very busy time in the breeding season and the island is coverd in grey seals of all ages, from newborns to females over 30 years old. The first weaners (pups who’s mothers have weaned them by going back to sea to eat fish, typically at about 18 days after birth) are starting to appear at the edges of the colony, without their mothers to protect them from aggressive adult seals the weaners quickly retreat to the peripheries of the colony. Even with so many seal pups of different ages around, it is still possible to tell approximately how old a seal pup is just by looking at it. It’s not always easy, but knowing the ages of the seal pups on the island is really important information for the various scientific projects studying the seals. For behaviour and physiology projects, it is crucial to know the developmental stage of a pup you are studying. For population dynamics research (which keeps track of the abundance of speices and detects changes in the numbers of individuals around our coast) knowing how long seal pups are present on a colony helps scientists accurately run models estimating pup production, an important variable in the model representing seal population changes around Scotland.

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The two pups on the right of this picture are clearly very different ages, but by how much? It’s important to know for all sorts of scientific research.

So how do you tell how old a pup is just by looking at it? Here is a crash course! Pups are divided into stages based on their appearence and the stages correspond to approximate ages (described here and drawn here). There are five stages in all, covering newborns to pups that have weaned and are independant from their mothers.

Stage 1.

These pups are the youngest on the colony, and this stage covers pups that are newborn up until about day 5 of life. Things to look out for include a pink, squishy umbilical cord from the pup’s stomach and pronounced shoulders protruding from the pup’s back as they are quite skinny when they are born. The back slopes distinctly away from the shoulders down towards the tail, and sometimes they have dried blood from birth in their fur.

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Comparison of stage 1 and 2 pups, the stage 1 pup on the left has a fresh pink, fleshy umbilical chord from it’s stomach and it’s shoulder blades are clearly visable. The stage 2 pup on the right has a dried up, black umbilical chord and has put on enough fat to stop the bones protruding from its shoulders.

Stage 2.

These pups are between 6 and 9 days old. Their umbilical chord has dried up and if it is still present, it looks withered and black. The pup will have increased it’s blubber layer just under the skin from drinking fat rich milk from it’s mother, so you won’t be able to see the shoulder blades easily any more. The back of the pup will still have a slight slope from the shoulders towards the tail, and there will still be a dip at the neck of the pup.

Stage 3.

Pups within this stage are between 10 and 16 days old and are substantially fatter than the previous two stages. The back of these pups is almost flat because of how much blubber is covering the lower abdomen, and these pups start to look a little like barrels. The dip at the neck will have gone, but the pup will still be entirely covered in white pup fur called lanugo.

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A stage 3 pup nursing from it’s mother on the Isle of May. The back only starts to slope at the pelvis, not the shoulders as previously in stage 1 and 2.

Stage 4.

These pups can be very distinctive because it is defined by the pup losing (moulting) it’s white fur. As soon as a pup starts to moult the white lanugo, it becomes as stage 4 pup. The pups usually start to moult around their faces and flippers first, then the rest of their body follows over a few days. If the pup rubs against rocks or other seals then the white fur can come out in large patches at one time, revealing the adult fur underneath which is often patterned with spots. These pups are between 17 to 24 days old. At day 18, mothers typically wean their pups so you can see ‘weaners’ with half moulted white coats around the edges of the colony.

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A stage 4 pup that has weaned from it’s mother and is resting at the edges of the colony. You can tell by the white fur on the ground around the pup that it has been in this spot a while! The black and grey fur underneath is the adult fur and the pattern will stay the same for it’s entire life.
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Three different examples of stage 5 pups, or ‘weaners’. Weaners can be all sorts of variations of grey, black and white including very pale (top), spotty (middle) or very dark (bottom).

Stage 5.

This is the last stage, and includes any pup that is more than 25 days old. These ‘weaners’ have no lanugo left on their body and can vary in colour from very pale, to spotty, to pure black. Weaners often stay on the breeding colony for several weeks after their mothers leave, which is called the post-weaning fast because they don’t have anything to eat while they remain on the colony without their mothers. To get food, they have to go to sea for the first time and learn to catch fish. This means that the amount of fat a pup puts on while it’s with its mother is very important for it’s survival; the fatter a pup is, the more blubber it has to burn for energy and the more time it has at sea to learn to catch fish before it starves. We still don’t really know why pups stay on the colony after weaning for so long as they often lose several kg of blubber during the post-weaning fast.

Being able to tell the age of pups is very important for the PHATS team as we are only interested in pups that are very close to weaning or have weaned from their mothers already (read our blog here for more details about why we sample the seals at these particular times). We have just identified our first study weaner, who is a stage 4 pup that has been dubbed ‘Charlie’ from the phonetic alphabet as her study code is ‘C’. We will be keeping an eye on her for the next two weeks during her post-weaning fast, so stay tuned for updates about her in the coming weeks!

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Introducing ‘Charlie’, our first study weaner! She weaned from her mother several days ago and you can see where she has moulted her white baby fur on her chest and flippers, while her back is still covered in lanugo (making her a stage 4 pup).

In the tissue culture lab on the Isle of May we have received several samples from pups that are about to wean from their mothers and have been successful in keeping the cells alive for the PCB and hormone exposure experiments we are carrying out. To make sure that the cells are alive during the experiments, we sent some of our tissue back to the PHATS laboratory at Abertay University back on mainland Scotland. To do this, Dr James Turton extracted RNA (ribonucleic acid) from the fat tissue in our experiments and determined it’s integrity by putting it on a gel and applying an electrical current to separate out the different sized molecules in the RNA (know as gel electrophoresis). RNA is a vital component of all living cells, it is how DNA (Deoxyribonucleic acid) in the nucleus of cells transfers information to other parts of the cell so it can function normally. If the fat cells died during the experiment, the RNA would degrade and would not show up on the gel. Thankfully our cells were all alive and showed clear white bands of intact RNA on the gel after the test, confirming the RNA has good integrity and the cell in our experiment were alive. Hooray!

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The RNA gel results from Dr James Turton showing clear bands of RNA molecules, which means our cells are alive!

The other research on the island is also progressing well, Dr Sean Twiss from Durham University has been working in collaboration with Dr Patrick Pomeroy from the Sea Mammal Reseach Unit at the University of St Andrews to trial UAVs (unmanned aerial vehicles) for use in seal research. As they fly high over the colony they have the potential to be able to see areas of the island that we otherwise could not study, however the high winds that often blow over the Isle of May can make flying the UAVs difficult! Read more about Dr Twiss’ research on his blog about seal behaviour here.

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The UAV flying over the Isle of May between the main light and the beacon.
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Fresh bread can be tough to come by on an island…
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Owl doodles, guess the species!

MEANWHILE away from the seals, the epic cooking and baking continues! I baked batches of fresh bread rolls for a monster hand-made burger night when it was my turn to cook for the masses. Dr Twiss engineered an incredible sushi evening for dinner over the weekend, almost too pretty to eat! Last night we had a stunning roast leg of lamb, which PHATS team member Holly was in charge of, so good.

Ouside of the kitchen I’ve been amusing myself by working on my sketching skills, doodling various birds that I’ve seen on the island in my sketchpad between all the lab work. Hopefully by next weekend I’ll have a whole menagerie in my book!

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Amazing sushi!

Isle of May 2016 – The PHATS team swings into action

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High winds and huge waves are no problem for seals hauled out far from the shore line like these two, but the seals that have pups within the tidal zone can quickly find themselves in trouble.
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The once grassy area south of Kirkhaven is now covered in breeding seals
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Crowded conditions often leads to more conflict between adjacent seals, this newborn is surrounded by other females and will have to rely on its mother to defend it.
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Mothers close to the sea may have easy access to water, but if storms hit the island they may lose their pup with the combination of high tides and waves.

Here on the Isle of May the grey seal breeding colony is getting crowded as we approach peak pupping time, which is usually in early November. As the colony fills up with adults and pups, there are more disagreements between neighbours or seals trying to move through the colony, travelling either to their pup or back to sea. So far this year it has been a dry season for the seals, we’ve not had much rain and the pools that are usually scattered throughout the colony have dried up. This unfortunately means that there is much more movement of females around the colony than usual as they try to commute to water sources and then back to their pups. Grey seals typically need access to pools of water, or the sea, regularly to help them cool down. The thick layer of blubber that keeps the seals warm when swimming in the cold north sea for days at a time means they can overheat when constantly out on dry land. In seasons when the weather is dry and pools are scarce, grey seals mothers spend much more time moving about seeking water and less time close to their pups, feeding and protecting them. For the seal’s sakes, I hope that we get a bit of rain soon even if it makes our fieldwork more unpleasent!

For the PHATS team, our project work within the field season is officially starting today. One of the study mothers has now produced a large fat pup who is only one day away from weaning, so last night it was time for research assistant Holly, team leader Dr Kimberley Bennett and myself to get our tissue culture lab ready for the arrival of our first sample. When a blubber sample is taken from a seal for our work we use it in as many ways as possible to get the most information out of the tissue. We run some of the blubber to detect persistent organic pollutants (POPs) contamination in the pup, which is what I was doing over the summer in Belgium with our samples from 2015 (read about that here). The rest of the blubber will be kept warm and well fed with media solutions containing sugars, salts, proteins and fatty acids so that the cells within the tissue stay alive. We can then expose these blubber cells to different experimental treatments of POPs (like polychlorinated biphenyls, PCBs) or hormones that are meant to regulate fat tissue function to explore how hormones control fat mobilisation and how POPs interfere with this process. Tissue culture works involves a lot of lab equpiment that is tricky to ship out to remote research sites, but we have managed to find a small CO2 incubator and UV hood which we can transport out by boat to the island. With a lot of ethanol and lab diligence, we can create sterile conditions for culturing cells out here for the PHATS project, and we’ll spend the next 6 weeks keeping the tissue culture lab running here.

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Our little UV hood sterilising after a deep ethanol clean, the UV light destroys bacteria or fungal spores that would love to get into the tissue culture plates and grow where our blubber cells are meant to be.
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Callan Duck’s aerial survey plane flying over the grey seal colony

Around the island all the other scietific research projects on both the seals and the birds continue as the season progresses. There are many other projects that are happening at the same time as ours on both the seals and the birds here on the island. Over the last week, Callan Duck from the Sea Mammal Research Unit has conducted an aerial survey of the entire Isle of May colony by plane to perform the annual count of grey seals on the island. Callan will fly over grey seal haul outs all across Scotland throughout the breeding season to monitor the size of seal populations around the country. The bird researchers from the Centre for Ecology and Hydrology are also doing the last mist netting and trapping to catch and ring birds on the island before their research station at the Low Light closes for the winter. They have been kind enough to show us many of the interesting birds they have ringed over the last week, inlcuding waxwings, fieldfares and a long eared owl!

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Long Eared Owl captured on the Isle of May by the Centre for Ecology and Hydrology for ringing just before release.
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Happy Birthday time!

MEANWHILE  We’ve had two birthdays in the last week so the PHATS team has been baking! I’ve made birthday cake, Holly helped make 60 profiteroles by hand and Kimberley made an epic beef and ale pie for dinner last night. The team leader here on the island also treats us to pancakes every sunday, so we had a lovely morning yesterday enjoying the biggest stack of pancakes I have ever seen.

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Pancake time!