S6 E7: It's Not Easy Being Green: The Story of Frog Fungal Disease

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I'm Dawn and I'm Cole and Scottish
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Because of Crime is a true crime
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focus on them murderers, so we
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past the true crime and scary science. Hello, and welcome to Cause of

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Death. One hundred seconds to midnight. I'm your host, Jackie Morante.

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Before we get started, I want
to welcome my latest patron to the family.

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Welcome, Elizabeth. I'm so happy
to have you here. I want

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to thank all of you for the
get well messages that you sent while I

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was sick. For those of you
who missed my messages, I had a

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little health mishap. What started as
a sinus infection moved into bronchitis, and

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then quickly ended up as pneumonia.
I'm feeling much better now and thank you

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for sticking with me. I apologize
that this episode is late, but I

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had to sleep it off for a
few days, so research wasn't happening.

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We're almost finished with season six Who's
Who Outside of the Zoo. After this

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there will be two more episodes.
Then I'm going to take a short hiatus

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and be back with season seven.
That season is going to be called Countdown.

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I have five or six one hundred
seconds to Midnight episodes in mind for

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that one. For those of you
who really like the one hundred seconds to

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midnight episodes, I hope you enjoy
that. I know I will. I

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really like doing research on those episodes. It's interesting to explore the topics that

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could change the world as we know
it. Today, we're talking about frog

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fungal disease. It's also known as
amphibian citridiomycosis. There is nothing like going

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to the river in spring, the
frogs are sending their mating calls back and

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forth. The males are trying to
sing the best songs to attract females,

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and they're measuring up the other males
to see if they really want to battle

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for the female they've chosen. When
you think about it, frogs aren't only

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important to ecosystems. They're important to
the natural rhythm of life. I thought

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that since we talked about white nose
disease and bats last time, we could

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just hop on over to frog fungal
disease this time. Let's begin with ideology

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and pathology. Batochochytrium dendrobades or B
D is a chitridiomcata. BD has two

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life stages, a motile zoospore and
a spherical, reproductive sessile zoosporangium. The

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motile zoospore directs itself and attaches to
the keratonized outer layers of its host.

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It will then absorb its flagella and
bury itself below the surface of the skin.

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In about four days, it matures
into a zoosporangia with rhizoids and produces

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and releases up to three hundred zoospores
into the environment via a discharge tube.

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Then the cycle starts a lower again. The fungus may retain the same host,

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or it may choose a different one. Frog larvae is thought to be

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a reservoir for the fungus, since
the fungus has been found in characterized mouth

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parts of frog tadpoles, but it
doesn't kill the tadpoles. Many amphibian larvae

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can survive for as long as three
years before metamorphosing. There's a lot we

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don't know about b D. It's
surmised that there may also be a symbiotic

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relationship between the fungus and some species
of salamanders and frogs, allowing them to

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be reservoirs of the disease. It
has been suggested that the fungus may be

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able to survive in non amphibian hosts
or feeding off of dead tissue until it

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can find a viable host. B
D is a diploid and primarily reproduces asexually

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and clonally by producing aquatic, uniflagellated
zoospores in a zoosporangium. Sexual reproduction of

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chytrid fungus has been associated with the
production of resistant, thick walled resting spores,

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and this leads the hypothesis that the
production of airborne spores could explain the

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widespread distribution of the fungus in relatively
pristine areas. Other research shows that the

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population structure of the fungus is largely
clonal, which would support the idea that

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the fungus lacks a sexual stage.
This gives rise to the thought that humans

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or birds could disperse the fungus over
long distances and bring that fungus into pristine

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settings. Citriteomycosis has been reported in
thirty eight amphibian species in twelve families,

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including ranid and highlid frogs, bufanid
toads, and plithontid salamanders. However,

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most of the population declines have been
confined to the montane rainforests. This fungus

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prefers cooler temperatures and grows best at
temperatures around twenty three degrees celsius, with

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slower growth occurring at around twenty eight
degrees celsius and no growth at all at

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twenty nine degrees celsius. Amphibian keratin
is used as a substrate for growth,

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and the fungus can grow for at
least one generation on cleaned epidermal keratin or

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on amphibians that have died of the
infection. A recent study by Voils and

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his team found that BD causes such
severe electrolyte imbalances in amphibians that it leads

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to an irregular heartbeat. Eventually,
their hearts just stop. The skin of

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amphibians encourages proper osmotic balance inside the
animal and regulates respiration. As the fungus

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takes over, the skin of infected
animals is less adept at transporting sodium and

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chloride ions, thus causing an electrolyte
imbalance. Sodium and potassium concentrations in the

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blood of infected frogs will drop and
more so as the infect and intensifies.

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Clinical signs of chitridiomycosis include acute death, anorexia, general malaise, and ventral

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edema. The skin may shed slaw
become arithmetis, display epidermal hyperplasia and discoloration,

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or become depigmented. Hyperkeratosis is associated
with diasdisis, and usually there will

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be evidence of secondary bacterial or fungal
infections. Primary lesions are generally limited to

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keratonized epithelial cells. Acute infections are
limited to the stratum corneum. In subacute

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and advanced stages, massive numbers of
Thallie will be found in the retain layers

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of the epidermis. Acanthosis may also
be present. This disease has very high

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mortality rate and has driven many amphibious
species to near extinction. Research suggests that

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the disease has spread globally due to
the importation of Xenopus species as pets.

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This species was imported from Africa and
many of these frogs were released in California.

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This introduced b D to naive California's
species and has consequentially wrought havoc.

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This is the only chichred fungus known
to infect vertebrates, and the host list

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is growing by the minute. Currently, it's considered an amphibian only disease,

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infecting both captive and wild animals.
This is a global threat, so if

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you're thinking of adding a new frog
tier, Amphibian habitat quarantine is important,

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as is disinfection of habitats and enclosures. Fungi as a rule love moist environments,

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so isolation of potentially contaminated animals,
enclosures, and equipment is vital help

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is on the way, though researchers
are working on cures, and of course,

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as we've learned in this season,
logistics are a large part of that.

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Wild animals are hard to treat,
and when you have a severe zoonotic

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event, it's a big deal to
get the cures to the animals, but

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it's a huge challenge to get the
cures to the right animals, and this

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case is no different. Several antifungals
work against b D, including voriconazole,

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but delivery and dosing are key.
The use of amphoteraesin b and vericonozole was

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published in Medical Mycology. At the
end of the study, it was found

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that amphoteraesin was toxic at the acute
stage in tadpoles, but vericonozole was non

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toxic even at high dose levels.
Vericonozole also remained stable in water, and

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after a seven day exposure period,
domesticated elites sister nazii frogs that had been

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sprayed with the treatment for seven days
were infection free. The experiment also included

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treatment of an infected colony of naturally
infected poisoned dart frogs, and that experiment

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was also successful. Several other treatments
have been suggested, but many of them

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would cause adverse reactions in the animals. Some of these treatments included soaking the

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animals in formaldehyde and malachite green,
raising the temperature in the enclosure to thirty

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seven degrees celsius, and treatment with
fluconazole and benzoiconium chloride. Any of these

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treatments would work, but they would
have long term adverse effects on the frogs.

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Itraconazole treatment would be successful, but
the animals would have to be soaked

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in the solution for at least five
minutes over a period of several days.

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Itraconazole isn't something that's readily available,
and this process would be very labor intensive

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and not easily delivered in the wild. For any of these treatment measures,

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the biggest problem is delivering these treatments
in the wild without affecting other species in

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the environment. That's the biggest problem
with any of the wildlife diseases that we've

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talked about this season. Treatment of
one species doesn't mean that you don't end

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up dosing the whole stream and the
fish don't need for a conozole. We

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all have bacteria that grows on our
skin and in our gut, and this

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bacteria serves as a barrier between us
and some of the pathogens that can invade

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our bodies. This is called a
microbiome. Frogs have it too, and

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while BD is resistant to most of
the bacteria that grows on frogs to protect

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them, there are a few that
have been studied that could confer resistance to

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the fungus. I'm planning a whole
season on the microbiome, but for now,

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I want to talk about two bacteria
that exhibit antifungal properties. Genetho Bacterium

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levidium in the family oxalo Bacteriasia and
Ceratia marcises in the family your Sinia asia.

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Researchers studied these two bacteria and found
that they produce antifungal metabolites biasin and

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protegeacin. Both comprimise the integrity of
the fungal cell membrane, and protegiacin can

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also inhibit RNA and protein synthesis in
bacteria. Several antifungal bacteria have been found

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in cold spots across the globe,
and it seems that the frogs who live

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in regions where BD is imprevalent to
have a higher degree of antifungal bacteria living

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on their bodies. This helps the
animals in three ways. First of all,

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they produce antibed metabolites. Secondly,
they produce a protective biofilm on the

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frog's skin surface, and lastly,
there is a host mediated selection for the

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antibed metabolite producing bacteria on the animal's
skin. Many of these bacteria live in

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the soil, so that can also
be a protective measure by not allowing the

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fungus to proliferate in the environment.
While research is ongoing, these studies offer

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some hope of a possibility of transferring
some of these antifungal bacteria to frogs living

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in high spots. Okay, we're
going to take a break to hear from

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my sponsors. Then we'll be back. This some history. It's the dog

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days of summer and most of us
are gathering the last of our corn and

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tomatoes from the garden. Soon we'll
be gathering pumpkins and squash for fall.

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Since I live in an apartment,
I have indoor garden beds and I grow

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00:16:22.960 --> 00:16:27.960
greens and herbs all year round.
This is why True Leaf Market is my

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go to when it comes to seeds, soil, and gardening supplies. I

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started twenty four herb plants this spring
indoors, and now some of my plants

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00:16:38.840 --> 00:16:44.720
are ready to harvest. I also
planted some greens in my garden beds so

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I can have those all winter long. Trueleaf Market carries everything you need to

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www dot true leafmarket dot com for
all your gardening needs. Don't forget to

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check out their flowers and cover plants. Frogs, salamanders and bats are attracted

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to many plants, so help our
keystone species. Visit Trueleaf Market at www

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dot trueleafmarket dot com today. Are
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with, True Consequences. Come by
and meet Eric Carter, Landein and I.

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We would love to meet you.
This disease existed for centuries in the

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Korean Peninsula, where it grew and
prospered on its own. Then sometime in

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the nineteen fifties during the Korean War, humans brought it to the rest of

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the world. Mass die offs were
noted in the nineteen seventies in the French

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Pyrenees, where the midwife frogs would
climb out of their birth legs and barely

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make it to shore before dying.
Biologist Matt Fisher described these events as quote,

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they'll do one last hop, then
they'll expire in your hands. You

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can walk the lakes. It's just
carpets of dead frogs end quote. Researchers

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didn't realize these dieoffs were related to
BD until nineteen ninety seven, when the

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connection was made. From two thousand
and four to two thousand and eight,

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a site in Panama lost forty one
percent of their amphibian species to b D.

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In two and seventeen, Jennifer Shelton
began exploring the mountains of Taiwan.

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She was looking for infected salamanders.
When anyone on her team found a sick

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amphibian, they snipped off one of
the animal's toes brought it back to the

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lab. Okay, before anyone gets
upset about the toe snipping. Snipping off

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a toe is a non lethal way
to get a skin sample, so after

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the toe was removed, the animal
could be released back into the wild.

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Researchers would then bring the toe back
into the lab and put it in a

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petrie dish to see what grew.
As the fungus grew, the researchers were

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able to sequence its DNA one hundred
and seventy seven b D genomes from around

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the world were combined with fifty seven
previously published ones. Two hundred and thirty

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four genomes revealed that there were four
distinct lineages in the family tree. The

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samples from the Korean Peninsula showed the
greatest genetic diversity of any of the other

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samples taken, and this is what
told them exactly what they needed to know.

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The Korean Peninsula is where it all
began. That was ground zero for

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b D. They then calculated the
mutation rate for the fungus and this told

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them that the ancestor of today's BD
line emerged out of Asia in the early

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twentieth century. This lineage is referred
to as the global panzootic lineage. A

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lethal strain called BD GPL. Human
activity is largely the blame for the global

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spread of BD. Shipping, live
frog pregnancy tests, the amphibian meat,

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and pet industries. They all gave
rise to a global zoonotic event. During

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the Korean War, frogs would hitch
rides on equipment being moved from one place

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to another, even if the fungus
couldn't survive without a host. A sister

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species of BD, known as B. Salamandra four ends, killed ninety nine

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percent of the Dutch fire salamanders from
two thousand and nine to two thousand and

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twelve. This spurred US Wildlife officials
to ban the importation of two hundred and

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one salamander species into the US in
an effort to keep B. Sal out

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of the country, but during a
US Court of Appeals ruling, it was

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decided that if the animals were already
in the country, their sales couldn't be

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banned, so they could and still
can, be transported anywhere in the US.

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Now, researchers are studying the inevitability
that less virulent strains of BD could

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come together and make a super virulent
strain of BD. This has already begun

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to happen a Brazilian strain of BD
mixed with the current virulent BDGPL, and

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the results were disastrous. It's been
considered that all amphibian trades should be banned

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globally, and wildlife conservationists are particularly
arguing this in the US. Currently,

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US Fish and Wildlife are monitoring the
spread of B D across the States,

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but they argue that since the disease
is already here. There is little that

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a ban would do to stop the
spread. Matt Fisher disagrees he would love

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to see a global ban on the
amphibian pet trade. Quote. Do we

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really need to be mining our environments
for organisms and selling them around the world

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for financial profit, just so we
can keep them in terrariums in our living

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rooms and say, hey, that's
cool. It seems like a fairly innocent

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pastime. But we are putting entire
ecosystems at risk. End quote. We

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are indeed putting entire ecosystems at Frogs
are a very important part of the world

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around us. Like bats, several
habitats depend on frogs for survival. Frogs

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are both a keystone species and an
indicator species in their environments. If we

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lose a keystone species, the entire
ecosystem will fail. Frogs provide food for

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several species, including birds, fish, monkeys, and snakes. As tadpoles,

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they eat algae, which helps regulate
blooms and reduce the risk of algal

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contamination. As they metamorphose from tadpoles
into frogs, they become more carnivorous,

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and this means that they eat a
lot of bugs. They can eat up

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to one hundred insects in a day. Okay, so this isn't quite as

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impressive as the bat, but still
that's a lot of bugs. The larger

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the frog, the larger the prey. Some species can eat animals as big

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as snakes and birds. Okay,
so maybe you didn't want to hear that

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part, but it does show that
frogs are an important part of the food

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chain as predator and prey. Frogs
are also an indicator species. If you

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listen to the Greed and Avarice episode
on chemical spills and the Environment, I

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talked a bit about researchers walking the
stream where the train derailment occurred in Ohio.

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They were looking for dead fish and
frogs. Frogs need clean habitats to

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survive. They need clean water and
clean soil. Anything that affects their environment

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will kill them. If the pH
goes too high or too low, frogs

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can't survive. This is what happened
immediately after the train derailment. There were

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many many frogs and fish found dead
after the event, and it told researchers

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and clean up crews that the water
was contaminated. People can't survive without clean

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water either. Go back to the
episode on the global water crisis, I

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talk a lot about how pollutants can
affect a water supply. If the frogs

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are gone, the water isn't safe
for anyone. That's what indicator species do.

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They tell us if our environment is
safe. Okay, we're going to

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go on to some trivia. Now. Back in the nineteen twenties, African

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clod frogs were used to see if
a woman was pregnant. The frog would

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be injected with urine from a woman, and if the frog produced eggs within

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twenty four hours, the woman was
pregnant. Today, the African cloud frog

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is still used in vertebrate embryology research. The females are prolific egg layers,

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and their eggs are transparent, so
it's easy to observe the growth patterns of

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the larvae. Epibatadine is a pain
killer that's two hundred times more potent than

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morphine. It's also the venom of
some poisoned dart frogs. People can't take

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ebibedodine for pain right now because it's
toxic to humans even in small doses,

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but research is ongoing to see if
there is a way to use it as

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a compound for pain killers. It's
a possibility that ebabatadine could be used topically

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when combined with another substance to relieve
pain. I'm going to take you back

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to middle school for just a minute. Remember when you were stuck in a

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biology class and the first thing they
made you do is dissect a frog.

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I know many of you got pretty
grossed out, but my biology teacher was

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pretty good. I remember looking at
the parts and pieces and trying to figure

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out how the body worked, especially
the muscles of the legs. That's what

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frog dissection was really about for me, learning how muscles worked, learning the

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attachments and what propelled the frog through
life. I learned a lot about muscle

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structure from frogs, and it was
my first look at a complex organ system.

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The dissection of frogs also gives us
insight into evolution. How does an

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organism go from living in water to
living on land? Frogs teach us that.

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Now. I'm not going to tell
you to go out and find a

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frog to dissect. Most of you
don't want to know how complex organ systems

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work. You just want to know
that they do work, and that's just

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fine. Leave the dissection to me. So I can figure out how to

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keep your organ systems working the way
they should. Thank you again for listening

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00:28:56.599 --> 00:29:02.720
to Cause of Death one hundred seconds
to Midnight. We have two more episodes

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00:29:02.920 --> 00:29:07.240
in season six, Who's Who Outside
of the Zoo. When this season ends,

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00:29:07.359 --> 00:29:11.640
I'm going to take a month off
to get ahead. That means I'm

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00:29:11.680 --> 00:29:15.960
going to be putting up more bonus
content. Also, I really didn't want

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00:29:15.960 --> 00:29:19.119
to take a break, but that's
the only way I can get a few

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00:29:19.119 --> 00:29:26.440
episodes ahead after having so much weird
life stuff that got me way behind.

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If you'd like to support the show, please take a look at the show

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00:29:30.960 --> 00:29:36.640
notes for links to my Patreon page
and my cofi account. You can also

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00:29:36.799 --> 00:29:41.640
sign up on Apple subscriptions for bonus
content and add free episodes. The show

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00:29:41.680 --> 00:29:45.960
notes are a great way to look
at some of the research I do for

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00:29:45.039 --> 00:29:49.880
each episode. If you're really into
white papers, or if you just want

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00:29:49.920 --> 00:29:53.880
to get your sleep schedule on track, go check out some of the links.

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00:29:56.079 --> 00:30:00.599
There's so much information on frog fungal
disease, medical search involving frogs,

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00:30:00.599 --> 00:30:07.000
and ways that you can help frogs
survive. Check out my sponsor for this

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00:30:07.079 --> 00:30:11.279
month, True Leaf Market. There's
a link to their website where you can

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00:30:11.319 --> 00:30:18.240
find gardening supplies and every manner of
seed. Summer is quickly coming to an

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00:30:18.319 --> 00:30:22.279
end, so now is the time
to think about growing indoors. I have

305
00:30:22.440 --> 00:30:26.960
my indoor garden beds all set up
and I'm looking forward to a winter of

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00:30:27.079 --> 00:30:33.400
fresh greens and amazing herbs. True
Leaf Market is my go to for all

307
00:30:33.440 --> 00:30:37.519
of my gardening needs. Go check
them out. The link is in the

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00:30:37.599 --> 00:30:41.680
show notes. I also want to
let you know that my daughter Jessica and

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00:30:41.720 --> 00:30:47.200
I are going to be launching a
new show in early October. It's called

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00:30:47.359 --> 00:30:52.279
Diving into Unsolved History. We're going
to look at some mysteries from the past

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00:30:52.400 --> 00:31:00.400
that are still unsolved, the Alien
More incident, the Ditluff Pass incident,

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00:31:00.079 --> 00:31:06.759
the Marie Celeste. We're going to
talk about famous treasures that could still be

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00:31:06.880 --> 00:31:12.039
out there, like Alaric's Treasure,
the Dresden Green Vault, Burglary, and

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00:31:12.240 --> 00:31:18.039
Thomas Beale's ciphers. We're going to
tell you the story behind the mystery,

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00:31:18.240 --> 00:31:23.119
look into the facts, and look
at some of the crazy theories that really

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00:31:23.200 --> 00:31:27.480
just don't make a lot of sense. It'll be a lot of fun and

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00:31:27.559 --> 00:31:33.440
I hope you join us, join
me next time when we look at human

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00:31:33.559 --> 00:31:38.519
encroachment on wildlife and the negative effects
it has on species. Thanks again for

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00:31:38.599 --> 00:31:41.279
listening, and have a great week.