Cities are accelerating evolutionary processes, with species adapting to urban environments within just a few decades rather than millennia; examples include moths developing reduced attraction to artificial light, mice evolving metabolic changes to digest human food waste, and plants producing heavier seeds to survive on asphalt surfaces, demonstrating that urban habitats create strong selection pressures that drive rapid genetic changes across diverse species worldwide.
Urban Evolution: How Wildlife Adapts to Cities | DW Documentary
Added:In a lot of ways you can think of cities as one of the largest unplanned experiments of all time.
Cities are places, we call them extreme habitats really, They are places, where there is a lot of opportunity and at the same time, there is also challenges.
As our cities spread, how will nature respond?
Will plants and animals dwindle, or will they adapt to urban life?
And what kind of new interactions will we see in the city?
In the historic French town of Albi, biologist Frederic Santoul keeps an eye on his catfish.
In 1983, fishermen released these Eastern European fish into the River Tarn.
Today, they’re at the top of the river‘s food chain.
This is a fascinating species because we know so little about them.
There are many myths that people believe, even that they eat dogs there are many stories.
The biologist is interested in the behavior of the large fish that circle the reservoir’s basins.
We work with fishermen to tag the fish.
They contacted us after observing very strange behavior in the fish here.
The manmade landscape of the city fosters new encounters between species.
The pigeons have never had to face predators from the water.
Instead they scan the sky for birds of prey.
The pigeons approach the water to bathe and drink.
Sometimes a bird misses the narrow strip of safe ground and touches down in open water.
The catfish don’t really see the pigeons.
But once they sense the birds’ movements in the water with their barbels - then they strike.
Scientists observed the catfish’s new hunting tactic for the first time in 2010.
Here in Albi, pigeons are no longer safe on the water.
The city is bringing together new predators and prey.
For some catfish here, pigeons now account for up to 40% of their prey.
There’s suddenly this ecological interaction, that allows for evolution to start.
To improve the bird-catching ability of the catfish, and also to improve the escape-ability of the pigeon, so you can expect that all these new interactions are also causing new evolutionary dynamics.
Dutch evolutionary biologist Menno Schilthuizen researches the adaptation of wildlife to the city.
Darwin’s theory, he believes, has gone urban.
Urban evolution is evolutionary change, so really genetic change in wild animals and plants in cities.
It's all about understanding how species will be able to survive in this very human dominated context.
Cities are Homo Sapiens' most extreme intervention in nature with concrete and steel we create new landscapes and alter the face of the earth.
Already most people live in cities, rather than in the countryside.
How does this influence evolution, the development of new species?
What selection pressures does the city create?
A summer evening in the Dutch capital Amsterdam.
In the Vondelpark in the center of the city, biologist Menno Schilthuizen uses a light trap to catch insects.
He is leading a 'Citizen Science' project to explore urban nature.
For insects and some smaller plants, the diversity today in cities seems to be higher, than in intensively managed agricultural areas.
Today, agricultural land is so intensively managed, and every last bit of production is squeezed out of every square meter of surface area, that there is no space for nature anymore in the countryside.
And at the same time, cities get more, they get greener, people pay more attention to nature and to urban nature, so it's actually becoming a very rich environment with a higher biodiversity, than outside of the city.
But overall, we’re rapidly losing biodiversity both within and outside our cities.
For insects, the declines are particularly severe.
In the Swiss Alps near Zurich, scientist Florian Altermatt has set up his light traps.
Ever since humans began to light up the night, billions of nocturnal insects have been dying off every year.
For a species like this, being attracted by light is problematic, because then it can’t use the few short days it has as a moth to lay eggs.
Light pollution is one of the major threats to moths.
Scientists are even going as far as to describe it as an “insect apocalypse”.
I think the declines we are seeing now are already quite worrying.
Studies show a 60 to 80% decline in biomass, sometimes even in nature reserves.
These are incredibly large numbers.
In my childhood I used to observe moths like these.
I would set up this trap next to my parents' house and attract moths, actually in quite large numbers.
But today I would probably not find many of them.
But might insects be capable of adapting to life in the perpetual light of our cities?
Florian Altermatt wanted to find out.
His test subject: the Spindle Ermine moth, whose caterpillars develop on the European spindle tree.
Actually, it was a coincidence.
While I was working on my PhD thesis, every day I walked through a park that had those European spindle bushes.
And I noticed there were these caterpillars, these moths, which must have lived there for years, in a city park with permanent light pollution.
I thought, I could just collect them, raise them and test how much the adult moths are attracted by light.
With his experiments in 2006, Altermatt pioneered research into urban evolution.
He released the moths in a darkened room.
The next morning, he counted how many had flown into the light trap.
The results showed a difference.
About 20% fewer urban moths had flown into the trap.
I was very surprised.
It was widely known that moths are attracted by light, some more than others.
But these differences have always been observed between different species.
Seeing variations within a SINGLE species that we’d never seen before.
The experiment clearly demonstrated a hereditary adaptation to life in the city.
Direct proof of urban evolution.
For Dutch biologist Menno Schilthuizen, the findings confirm a larger picture.
In Amsterdam, he and his group of citizen scientists debate whether we might soon observe even more and greater adaptations of animals and plants to the city.
We see that evolutionary processes are starting, which will eventually or could eventually produce new species that are specialized to living in the city.
For Menno Schilthuizen, it’s not if, but when.
Every organism that lives in the city will show this urban evolution, these rapid changes in their behavior, in their physiology, in their appearances to optimize their life in an urban environment.
But what elements of urban landscapes prompt wildlife to adapt?
Evolutionary biologist Jason Munshi- South is an expert on animals found in the parks of New York.
For years he has been studying how rodents adapt to the city.
Along with human immigrants from Europe, rats also voyaged to the New World.
Today they roam the city in subway tunnels.
Most native rodent species, however, don’t dare try their luck crossing town.
This distinction sparked the scientist’s interest.
I used to be a tropical biologist.
But then I moved to New York City for my first academic job after graduate school, and I decided I wanted to do some local work, that would be interesting to the people of New York City, and to myself.
And I found out that there were these small mammals, living in essentially islands of forests in the city.
And I thought: that is interesting nobody has ever really looked at these.
Are they becoming genetically different from mice outside the city, or are they adapting, and that’s how it all started.
Central Park opened in 1873.
It still hosts animal species that lived here long before the city was built.
Right now, we’re in the middle of Central Park.
We’re gonna be travelling to the north end of the park, where there is a very nice forest called “the North Woods”, and there we’ll be setting up traps, hopefully to capture white footed mice.
One of the things that inspired me, when I first started this work, is if you look at a New York City subway map, you see the subway lines, but then there are these large green shapes rectangles and ovals and so forth, that are the parklands.
And they put those on the map, so that you know where they are.
But you also see that they are almost like a chain of islands, that are scattered in a sea of concrete and roads and buildings, you know, 8.5 million people.
So in a sense, if it's a species like a mouse, that can’t leave the forest, cross you know, neighborhoods, and buildings, and roads and make it to the other patch, it is essentially the same, biologically, as if they were on an island.
In terms of them not being able to move and spread their genes between the other patches.
And these urban patches, once they become sufficiently isolated, operate like a mini Galapagos, and maybe driving the evolution of many species, that are stuck there now.
The evolutionary biologist is investigating whether the white-footed mice actually develop in distinct ways in each of the various parks.
So this would be a really nice spot for white footed mice.
They like to move next to logs, so they’re not completely out in the open, they might actually be even living inside this log, or in holes underneath the log, so this is pretty much the ideal spot.
This forest is encircled by the Big Apple.
Have the mice already adapted to this unique environment?
What traits do they need to survive here?
No shortage of goog trapping spots Later I’ll be going to one of our more suburban, almost rural sites, with a larger, more intact forest, less urbanization, and I’ll be setting out you know, an equal number of traps, you know, with the hope that we’ll catch mice there as well.
Jason Munshi-South will search within the animals’ genetic codes for the markers of life in the big city.
I think what's been most interesting to me, is thinking about how the things that we are all doing in our daily lives where we put our garbage what we're choosing to eat and what we generate as waste, where we choose to live, how we choose to go to work or out to a restaurant or something.
All of these things we are doing are now influencing other species in a way that we're just starting to understand.
But it’s not only animals that adapt to human intervention in the natural world.
Plants do the same.
In southern France, the yellow-flowered "Crepis Sancta" is being studied by biologist Pierre-Olivier Cheptou.
Crepis Sancta is a very common species in the Mediterranean region, a kind of Mediterranean dandelion.
From the same family.
And its essential advantage as a model is that it produces two types of seeds: large ones and small ones.
The small wildflower produces both lighter seeds with parachutes allowing them to glide, and heavier seeds that simply fall to the ground.
I'm interested in the process of adaptation to an urban environment and in particular what happens when a species first arrives in the city.
It has recently colonized certain areas of Montpellier.
In my comparison between rural and urban populations I focus on the traits related to seeds.
The idea of studying the adaptation of the modest plant to the city came to Cheptou almost by chance.
When he came back from abroad, he noticed the inconspicuous flowers growing in the city.
I had left Montreal in the middle of a blizzard.
I took the plane for Paris.
Then I took the bus to downtown Montpellier, where it was sunny with a clear blue sky.
And then I noticed there were crepis sancta flowers everywhere in those tiny urban patches.
And suddenly it stuck me that there was something to figure out here, but I didn't yet know exactly what.
Because actually, the crepis sancta thrives in rural areas, and not in the asphalt deserts of the city.
The predominant component in cities, especially in European cities, is concrete.
Concrete exerts a powerful, fragmenting force on the habitats of plants they have to survive in mini-biotopes Sometimes the city's constraints on a plant's habitat can be extreme.
How will evolution respond?
I‘m looking at how urban fragmentation will modify the dispersal traits of this species.
I expect plants that produce more of the larger seeds will be more successful at reproduction in urban areas than they would be in the open country.
The heavier seeds are less likely to be swept onto the asphalt.
And indeed, the biologist discovered that far more plants in the city produce the heavier seeds and are thus better able to survive a difference of 15%.
But what stands out most is the speed of this adaptation.
The evolution we have seen has taken about 15 years.
This is extremely brief.
It was the first demonstration of such a rapid evolution of seed traits for plants.
And this is due to the highly fragmented composition of the urban environment.
Genetic changes occurring at such a rate have long been considered unlikely — even impossible — by science.
I think Darwin would have been amazed by the fastness by which these changes take place.
He was sort of underestimating the power of natural selection himself.
He said you can never see any of these changes in progress.
You cannot actually observe them, you can only deduce them from the fossils from the patterns that you see in nature.
He said evolution is too slow to see it happening in real time and the fact is, that now especially in cities we see this changes taking place under our eyes, in the streets where we live, right around us.
I think Darwin would have been thrilled by that.
But what if manmade pollutants substantially distort the biochemistry of organisms?
In the 1970s, the water at New Bedford Harbor, near Boston was severely polluted with PCBs.
The US Environmental Protection Agency wanted to know just how bad the pollution really was.
The original focus was on what must be wrong with all the fish, that live in that harbor, because of the toxic chemicals.
Instead we came here looking trying to understand, what must be right about those fish, that can survive here.
So they’ve become a natural experiment for us to study, how animals can adapt, to toxic human made pollutants.
Terrific, just what we’re looking for.
Let’s get them into a net and bring them back to the lab.
Diane Nacci heads the Environmental Protection Agency lab in Narrangansett, Rhode Island.
In their breeding facility, the scientists want to unravel the mechanism that allows this population of a killifish species to survive in the PCB-polluted water of New Bedford Harbor.
Let’s see if they left any eggs for us.
They plan to compare eggs from the New Bedford Harbor fish with those of a fish population from a cleaner site.
Ok, let’s start a test and see what they do when we expose them to chemicals.
This killifish species occurs all along the North American Atlantic coast.
The Killifish has been a favorite of biology, literally for centuries.
They are quite common, they are non-migratory, so they reflect their local environment, and each population is unique in that is genetically different, it is adapted to its local environment.
So it gives us opportunity for lots of studies.
The researchers need to observe the development of the fish embryos in the egg in order to understand at which stages the environmental toxin disrupts the animals' biochemistry - or not.
So we’ll look at the rate at which the embryo is developed, and certain features, that we know that PCBs can disturb, like a proper development of the heart, evidence of proper development of the circulatory system, and proper body-size.
Why are these particular fish able to resist deadly environmental toxins?
What are the factors that allow individual species to adapt to the city?
Are parallel developments taking place in cities worldwide?
At the University of Toronto Mississauga, evolutionary biologist Mark Johnson pursues these questions.
In a lot of ways you can think of cities as one of the largest unplanned experiments of all time.
The problem is that there are very few organisms where you could study adaptation to urban environments on a global scale.
And white clover is one of those very few organisms where you can actually do that.
So now, this becomes then the model, to understand whether organisms in general, can adapt to the convergent environmental changes associated with cities throughout the world.
Researchers across the globe are working together in this study.
Evolutionary biologist Stephan Greiner and his team are collecting the white clover in Berlin.
In cities, the plant faces a different habitat.
Temperatures are higher than in the suburbs and the countryside.
What you can expect is that as humans create new environmental conditions, life will adapt.
And to be able to show that on a global scale, that's a real scientific benefit.
And that is why we’re dedicating our free time to this project.
As they proceed from the countryside to the city center of Berlin, Greiner and his team collect specimens at 35 locations.
This gives them a sufficiently broad range of data to compare with that of other global cities.
They find their final samples at the foot of the Television Tower.
That's it. All done!
In all we have 168 cities right now and over 250 collaborators all working on the same project together.
There’s never been a collaborative project on evolutionary biology of this scale.
So this is the largest collaborative project in evolutionary biology ever.
So is clover developing in the same way all over the world into a kind of "global city clover"?
From the vast set of data, the researchers hope to find an answer.
In the grounds of a research institute north of New York, geneticist Jason Munshi-South wants to catch white footed mice to compare their DNA with that of those in the city.
But it’s not easy.
So this is a trap that was opened that did not catch anything obviously.
Oh, that’s a toad.
I thought that was a mouse.
It’s nice, a day for other wildlife, I guess.
We were really surprised to see that almost every park was different from every other park.
It's almost to the point where you can take a mouse from one park, give it to our lab, and we could just look at a small segment of its genome, and tell you, where it came from.
That's how much they had changed just randomly over time from being isolated.
And that’s when we started our current studies, looking at, you know, over 20,000 genes, to see what genes and potentially what functions change when they adapt, to living inside of New York city.
Oh, there’s one.
First white footed mouse of the day.
For the new study, Jason Munshi-South and his team have already caught more than 100 mice and analyzed their genetic compositions.
They try to take their samples as gently as possible so as not to hurt the animals.
We take a genetic sample, in this case, we’ll be using this small tool, it's like a paper punch but for tissue.
And we store that for genetic analysis.
We want to be able to tie that tissue sample to a location, because that is important for understanding how they vary, when they’re in a more urban or less urban population.
So, now they’re pretty immobile.
This is a male, this is a young male.
Why don’t we take the ear punch, and start on the other one.
Hair isn’t suitable for comprehensive genetic analysis so the researchers take a tissue sample from the ear.
After collecting the samples and some measurements, the scientists release the mice.
Genetic analysis can reveal the evolutionary trajectories of the mice.
They point to a variety of physical and behavioral changes spreading among the animals.
Each of them unique to the challenges of each city park environment.
So we’re starting to fill in our gradient really nicely.
So here are the mice we have today from the Calder Center.
You can see its right in between highly urbanized New York City and then all these sites we have up here, and one out here.
So central park seems to be our most distinct population today It makes sense!
It makes sense, yeah, most urban, probably the most isolated.
So if you took a mouse from Central Park, some of its genes will be different from a mouse from outside in the countryside in a big park somewhere.
The food supply in Central Park — much of it human food waste might have triggered a genetic response.
So what we’ve learned so far is that one set of genes that are changing in the city have to do with metabolism.
So these white footed mice are eating things, and then they have to digest them and assimilate the nutrients.
And we know its evolution, because inheritable change in DNA-sequences is evolution.
Central Park mice seem to have genetically altered their metabolism to better digest fast food.
It raises several like broader questions, about what we’re doing as a species, as we modify the earth’s habitat for our needs.
How are we changing the future of other species?
Not only are we affecting them but we’re changing what they will become in the future.
In Narrangansett, Diane Nacci and her team are investigating how the different fish embryos exposed to the toxic PCBs have developed.
Hi, how is it going?
Really well so, this is the study that’s comparing Scorton Creek and New Bedford harbor, exposed to PCB 126.
So, this one is the group, that was treated with PCBs, also, from the clean site, at Scorton Creek.
And, as you can see, the PCBs have had a pretty dramatic effect on the development.
Which is what we expect with these very toxic chemicals.
Yeah, and in my experience, when I see this, this constellation of anomalies, it's absolutely lethal.
There is no way, that an animal would even hatch, never mind survive after hatching.
If the heart is not functional and the blood is essentially not circulating around the body.
So let's take a look at that biochemical endpoint, to see if they’re also responsive, at the biochemical level.
Using a special contrast agent, the scientists can trace enzyme activity in the unhatched fish.
So you can see the substrate is fluorescing in the bladders, showing that this enzyme system is working and we’re getting the expected metabolites in the bladder.
That’s a very dramatic demonstration of enzyme activity in a living organism.
The active enzymes in the fish embryo reveal how the organism tries to break down the toxin but perishes in the process.
Then the team observes how the offspring of the fish from New Bedford Harbor have developed.
OK, so these are fish from new Bedford Harbor, that were exposed to the same level of PCB that we were just looking at.
As you can see with this embryo, it doesn’t seem to have any effect.
The heart is still beating normally, and healthy and its developed really well.
That looks like an embryo that’s about ready to hatch.
Some of them actually already hatched.
These fish should be dead, poisoned by one of the most lethal environmental toxins.
But life, it seems, has found a way.
One thing we know about this class of chemicals is that in all vertebrates, including people, it turns on a certain enzyme pathway, so a normally responsive person, or in this case a fish, should have that enzyme system turned on, if they were exposed to PCBs.
The contrast-enhanced image shows how the enzymes that normally respond to the toxin remain silent.
So in this case I see very little that’s glowing brightly, it is a dramatic visual difference, that suggests that that enzyme system is broken, in the New Bedford Fish.
The killifish from New Bedford Harbor have changed their metabolism the poison can no longer harm them.
But which genetic modifications lead to the fishes’ toxin resistance?
That's what geneticist Mark Hahn of the Woods Hole Oceanographic Institution wants to find out.
Could this be a key to understanding how nature might resist human interference?
In the laboratory he uses the CRISPR-CAS method.
It is an extremely powerful way to modify the genetics of an experimental fish like this.
To ask questions about the roles of certain genes, and in fact the roles of even single amino acids in the protein can be investigated with this CRISPR-CAS method.
To test their assumptions about the resistant Killifish, Mark Hahn and his team experiment with Zebrafish.
I wanna find out what exactly are the changes in those genes.
And to be able to actually zero in on the specific molecular changes, that are responsible for the resistance, and to be able to recreate that in the laboratory, to actually prove, that that’s the mechanism of resistance.
They’re inserting portions of DNA taken from the resistant Killifish into embryos of Zebrafish.
Hey Mark, how are you?
Good!
Which ones are these?
I am injecting AIP Exon 2.
Mark Hahn and his colleague Neel Aluru are using the most up-to-date genetic engineering techniques.
Here we are interested to study a function of a gene known as AIP.
So we’re trying to delete this gene in this particular species, and try to study, what’s the function of this gene and whether that will alter the resistance to PCBs.
With these experiments, science is venturing deep into the source code of creation.
The scientists believe this research could yield the secret of life‘s ability to adapt to the most extreme conditions.
And this knowledge could also help other creatures to adapt and survive in a rapidly changing world.
I think we will understand, the extent to which we can extrapolate our knowledge from the killifish-system out beyond to other fish and even other vertebrates.
So, a broader understanding of the toxicology of pollutants and how that will impact the natural world.
How we can understand what will be the most vulnerable species.
At the Max Planck Institute in Potsdam, the research team processes the clover samples from Berlin.
Their goal? To find cyanide.
Clover plants that produce cyanide are better protected against predators but are less able to tolerate cold.
It’s warmer in city centers, so this clover might be more common there.
This is a qualitative test.
We use it to indirectly detect a specific gene that generates this cyanide.
They're both rural now.
Yeah, they're both still rural.
Greiner and his team send their results and other clover specimens to Marc Johnson in his Toronto lab.
Hi Gang, How is it going?
Good.
How did this extraction go yesterday?
Good!
So remind me, this is Berlin and Buenos Aires.
The team prepares the clover for gene sequencing.
But the cyanide values taken by the team in Germany already show weather the clover has adapted to an urban existence already.
Ok, did we get the data from Berlin.
Yes, we did.
On this screen here it is, So I think it is really good.
James. Did you have the chance to look at the data from Berlin so we understand how the environment is changing from downtown Berlin through the suburbs and the rural areas.
You got it.
So, Berlin is one of the cities where we see white clover adapting to urban and rural gradients.
Yes!
So now we are at 33% of the cities where white clover adapts?
Maybe at 33.2% right now But, yeah, it is at about 33% So then next I think is what we have to do, is to figure out, what are the drivers, the environmental drivers of this adaptation.
So that's really cool!
In Berlin, plants from the city centre are more likely to produce cyanide as is the case in a third of the cities surveyed so far.
An indication of parallel evolution?
Some of the preliminary insights are fascinating.
It really looks like, regardless of where you are in the world, whether you are in Europe, North America, Japan, China, Australia, New Zealand, we see the ability for this humble white clover, to adapt to these cities.
In the warmth of the city, cyanide-producing clover stands a better chance of survival.
But to survive in the city, all organisms must adapt to higher temperatures what scientists call “heat islands”.
In cities, humans and their machinery create a lot of heat, and we have a bubble of hot air in large cities.
and a city of more than a million people can be 7 or 8°c hotter in the center of the city than outside of the city.
This - Menno Schilthuizen believes also influences the evolution of the White Lipped Snail.
Their shells come in many shades, from brown to pale yellow.
A single gene determines the color.
So they basically carry their genes on their back.
The shell color determines the internal temperature of the snail, to some extent.
The difference in temperature inside can be 2° under the same conditions and that could be just the difference between life or death.
On a hot summer day, and you know it was 40° in Amsterdam a few weeks ago, it could be that some of these yellow snails survived, but many of the brown ones died, because they got too hot, they overheated, and they died.
But will the statistics support this hypothesis?
So the plan is now to just add some data to the dataset.
So let's go.
In order to collect and evaluate as many snails as possible throughout Europe, Menno Schilthuizen Schilthuizen is helped by volunteers.
You know, you don’t have to go to the Galapagos to study evolution, or become a paleontologist.
It’s happening everywhere all the time, it's a continuous, very normal biological process.
The group only finds a few snails, but even empty snail shells can also provide data.
They’re photographed and added to the database with an app that anyone can install on their mobile phone.
So we’re looking at the adaptations of urban animals and plants to the urban heat island, which of course has been happening more rapidly, than global climate change, so we can probably predict, what’s going to happen globally, in response to climate change.
Urbanization and climate change pose a threat to all plant and animal species.
Including the Monarch butterfly, which gather in their millions in the forests of Mexico every October.
They’ve completed a 5000 km journey to their winter quarters.
An increasingly perilous odyssey for the insects.
Lindsay Miles studies the butterflies in Toronto.
Monarch Butterflies are these really great insects.
Unfortunately right now they are in decline.
In the United States they’ve experienced 80% population declines.
In this industrial area of Toronto, Monarch butterflies take a rest stop before flying on.
They also take the opportunity to mate and reproduce.
There is a really big caterpillar... So this is a baby monarch this one is probably a day or two away from going into its chrysalis and then becoming a monarch butterfly.
While other species have the ability to switch to other food sources, Monarch butterflies remain dependent on a single plant.
Situative student I got one Let’s check it out Wow, you found a Monarch.
The butterflies lay their eggs on the Milkweed their caterpillars feed exclusively on this plant.
And in many cities, the land on which Milkweed can grow is disappearing.
Unfortunately a lot of these cities are providing these barriers and just don't have the resources that they need.
So it would basically be: if you are driving along a road and you don't have any fuel stations and you run out of gas.
You’re stuck and that’s what’s happening with these butterflies.
Not all species can adapt.
As our cities continue to expand, accommodating wildlife might be crucial.
How we shape our cities in the future may prove decisive for the course of life on earth.
Biodiversity helps us with the food that we eat, it helps us with the air that we breathe so if we continue along the path that we have many populations including human populations will start to crash.
Urban evolution can help us to design green cities in a Darwinian way.
As humans become more urban, we have the potential, to, you know, allow some species to live in the city, and adapt to our cities, but then, put less pressure on the other habitats.
Which will allow the species that can‘t survive in the city, to continue to thrive.
We are going to see more and more the realization that we are part of nature.
And that is actually probably going to help us survive.
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