Showing posts with label toxicofera hypothesis. Show all posts
Showing posts with label toxicofera hypothesis. Show all posts

Wednesday, April 10, 2013

Sneaky Snakes: Biters, Boobytraps, and Spit

Biology concepts – venom, toxin, poison, fangs, evolution, toxicofera hypothesis

The California Red Sided Garter Snake is one beautiful
reptile.  It has round pupils, so it might be non-venomous,
but it has a highly patterned and bright colored body, so
it could be poisonous. It has a triangular head, but not to
broad, so it could be either. For many years we thought
it was non-venomous, but recent work says it has toxic
saliva. It’s probably not too harmful to humans, but it
goes to show that you can’t make assumptions.
If you know anything about this blog, you know that we look for exceptions in every rule, so I wouldn’t tell you that there are definitive ways to tell venomous from non-venomous snakes. There are just too many exceptions; any mistake in this area could be your last.

However, many people will tell you that the eyes of a snake will give them away –round pupils means non-venomous, while slit pupils (like cats) means venomous. Or that venomous snakes have patterned bodies while non-venomous snakes wear solid colors. Lastly, some people will tell you that venomous snakes have triangular heads, while non-venomous snakes have rounder heads.

Let’s tear down each myth. About 99% of snakes have triangular heads. So this is no help at all; although, if you stay away from all triangular headed snakes you probably won’t get in trouble. Venomous snakes do have a broader base to their triangular head, to account for the venom gland volume and associated muscles. However, are you going to take the time to determine just HOW BROAD is the head of the snake that’s about to bite you?

As for pattern versus solid color; this will also fail you. Ever hear of a black mamba (Dendroaspis polylepis)? Well, it’s the most venomous snake in Africa and it’s solid colored. Strangely enough, the black mamba isn’t black. Its name comes from the color of the inside of its mouth, but its body is silvery. The Inland Taipan viper (Fierce Snake, Oxyuranus microlepidotus) is the most venomous on land, but it has a solid dark tan body.

Maybe pupil shape matters. I am wondering why there would be an evolutionary link between the shape of the pupil and whether a snake has venom. Head shape – maybe, you have to account for venom glands. Body color – maybe, patterns would warn a predator to stay away (aposematism). But pupil shape? How would that be linked to venom or no venom?

The black mamba is one of the most aggressive of venomous
snakes. It’s as if it goes out looking for things to bite. And
they are fast, moving up to 20 ft./sec (that’s 14 mph, or 22.5
kph), although I think Kobe Bryant can still move faster than
that in short spurts. The mamba grows to over 10 ft in length
and will attack a lion, so it backs up the deadly look of its
open mouth – it ain’t just for show.
We can use the black mamba and inland taipan snake examples for debunking the pupil myth as well. The mambas and the fierce snake are round, as are the pupils of the coral snake (very dangerous). More to the point, who wants to get close enough to a snake’s pupils to see if they are slit shaped or round? Let’s err on the side of caution; if you see a snake in the wild – assume it’s venomous. Problem solved.

So let’s find some real exceptions in the realm of venomous snakes. Unfortunately, there are so many different combinations of fang type, venom type, and venom gland type that it is difficult to call any kind of venomous snake an exception – there aren’t many rules. There are true venom glands and false glands, based on whether they can store venom. Then there are rear fangs, front fangs, and front fangs that can fold up. And then there are systems that deliver venom to several upper teeth, and those that deliver venom only through the channels in front fangs or rear fangs.

There may be different ways to deliver and different venoms to deliver, but a 2008 study says it all venomous snakes derive from a common ancestor that lived about 60 million years ago. The study of Dr. Vonk looked at front and rear-fanged venomous snake embryos, and saw that the venom gland ducts ALWAYS start out attached to a rear tooth, but in the front-fanged snakes, the tooth and duct move forward during fetal development!

The rear-fanged snakes have back teeth that deliver venom,
but not by injecting it like a hypodermic. They are not hollow;
some have grooves for the venom to run down, while others
are just sharp, bigger teeth. Snakes with rear fangs have to
take a bigger bite, and hold on longer to grind the venom into
the wound. Doesn’t sound like the perfect system, but
evolution is striving for perfection, it just uses what is there.
What is more, it would seem that the tooth that developed into a fang became linked to the development of the venom gland rather than the other teeth, suggesting that they co-evolved in all types of venomous snakes, linking the type of venom with the type of delivery and type of fang. Perhaps they all started out the same, but they developed their own combinations independently.

It may be that this occurred with all snakes; those that aren’t venomous just lost the ability to produce or deliver venom. This is part of the toxicofera hypothesis of which we have spoken. We don’t even know what percentage of snakes are venomous. Scientists have focused on the highly venomous snakes for so long, that the so-called non-venomous snakes have been ignored.

Many snakes that were once thought to be non-venomous are now known to have venomous bites. Colubridae snakes (garter snakes, hognose snakes and many more) were thought up to the 1950’s to be utterly non-venomous. But Dr. Fry showed that many of these snakes do indeed deliver venom, though most may be harmless to humans.

As recently as ten years ago it was said that only 10% of snakes were venomous; now that percentage is somewhere near 30%. Where might it end – could most snakes be venomous?

Even though scientists now know more about the evolution of venom, there are still mysteries. With millions of years to perfect a venom system, why is it that some snakes have venom that is WAY TOO POTENT for its purpose? With each bite the fierce snake delivers enough venom to kill 2000 mice or 50 humans – why so much? It must be advantageous in some way; or else it doesn’t cost any more energy to make the venom that potent.

This is a juvenile tiger keelback snake. The raised part on the
back explains the name. It is actually the nuchal gland that
stores cane toad toxin as a defense. How could a small
juvenile have toxin before it is big enough to start eating cane
toads? It can be passed on from mother to offspring, if she
ate a cane toad before egg format
Great diversity in venoms and fangs aside, there are exceptions in venomous snakes. Let’s talk about the keelback snakes. They may be venomous, but they are also poisonous.

Debra Hutchinson published on the keelback snakes in 2007. Tiger keelback snakes (Rhabdophis tigranus) live in Asia, and enjoy a diet of cane toads – poisonous cane toads! Cane toads kill most things that try to eat them, but for some reason the keelback snakes don’t seem to be bothered by the toxin.

In fact, they sequester the cane toad toxin to two nuchal glands, located on the back of their necks. Then, when the snake is threatened by a predator, they turn their back to the aggressor and dare them to bite down on the nuchal glands! Most predators have learned not to take the bait.

The nuchal glands are purely for storage. They don’t have ducts or deliver the poison to the skin or a fang. The sequestered toxin is purely defensive. But the keelbacks also have venom glands, of the false gland type, delivered to the base of several upper teeth. The keelback then bites, chews and grinds the venom into the wound.

This is the way it goes for many rear fanged snakes. Delivering venom by the front fangs is 100x more efficient than the rear fangs, so rear fang snakes must hang on longer to their target in order to envenomate them. This means that they are more vulnerable to being bitten when the target fights back. The keelback has made this less likely by storing another toxin in its neck. Pretty smart, huh?

When threatened, the tiger keelback assumes a familiar neck
arch position. This was described in Akira Mori’s 2012 paper.
If disturbed, it will also try to bump the nuchal glands into the
aggressor, They will spray toxic contents if bitten or pinched.
Now for one more venomous snake exception. Your parents always told you not to spit, but a few snakes are expectorating geniuses. The spitting cobras (genus Naja, and a couple others) have an additional modification to their front fangs that gives them the ability to spit their venom, in some cases, over twenty feet.

Injecting venom from front fangs is controlled by specific muscles around the venom gland. Spitting snakes combine this quick delivery under pressure with a targeting system. Instead delivering venom from the tips of their fangs, they have an aperture (hole) in the front face of their fang (see picture). Some cobras aim for the eyes of their targets, while others aim for mouth, nose or skin.

The aim is incredible in all, but it is even better in some species. I will use guns as a model. Most guns and cannons up to the time of the US Civil War were very inaccurate. By gouging curved grooves down the barrel, a spin was placed on the cannon ball, and the spin made it much more accurate. This “rifling” was invented in the 1500’s, but didn’t become common until the 1800’s.

The same is seen in the African (not so often in the Asian) spitting cobras. The fang and aperture have rifling grooves that make them even more accurate. I would say that humans stole the idea from nature (like we so often do), but I don’t think we knew about spitting cobras when guns barrels started being rifled.

But how does spitting (really squirting, no saliva is involved) venom at an aggressor help, other than grossing them out? We know that venom must be injected below the skin in order to be effective, but a spitting cobra’s toxin can be cytotoxic (lots of inflammation and tissue destruction) to the skin, and can blind if it hits the eyes. The black-necked cobra and the red Mozambique cobra have been shown to aim only for eyes.

This is a Mozambique spitting cobra. Notice how the spray
comes straight out from the front of the fangs and is directed in a
narrow, pointed direction. This isn’t strafing fire, it’s sniper work.
In a 2005 study of spitting cobras, the red and black-necked spitters recognized faces and eyes, but would not spit at photographs of faces. That’s mighty evolved; you don’t want to waste the toxin, and you must be accurate to avoid waste as well. The black-necked was able to hit the eyes 80% of the time, and the red spitter never missed.

Most spitting cobras actually have a mix of toxins; some neurotoxic, some hemotoxic, some cardiotoxic, and some cytotoxic. Somewhere along the way, evolutionarily speaking, the spitting cobras concocted a toxin that has both the ability to harm by surface contact, and the ability to harm on contact. Evolution at its best.

Next week, is there a group of animals where every species is venomous? And how about a group where none of the species are venomous.


Vonk, F., Admiraal, J., Jackson, K., Reshef, R., de Bakker, M., Vanderschoot, K., van den Berge, I., van Atten, M., Burgerhout, E., Beck, A., Mirtschin, P., Kochva, E., Witte, F., Fry, B., Woods, A., & Richardson, M. (2008). Evolutionary origin and development of snake fangs Nature, 454 (7204), 630-633 DOI: 10.1038/nature07178

Mori, A., Burghardt, G., Savitzky, A., Roberts, K., Hutchinson, D., & Goris, R. (2011). Nuchal glands: a novel defensive system in snakes Chemoecology, 22 (3), 187-198 DOI: 10.1007/s00049-011-0086-2
 
Hutchinson, D., Mori, A., Savitzky, A., Burghardt, G., Wu, X., Meinwald, J., & Schroeder, F. (2007). From the Cover: Dietary sequestration of defensive steroids in nuchal glands of the Asian snake Rhabdophis tigrinus Proceedings of the National Academy of Sciences, 104 (7), 2265-2270 DOI: 10.1073/pnas.0610785104


 

For more information and classroom activities, see:

Snakes –

keelback snakes and nuchal glands –

spitting cobras –

Wednesday, March 27, 2013

Hang A Question Mark On It

Biology concepts – toxicofera hypothesis, evolution, venom, toxin, reptile

Bertrand Russell lived in England in from 1872 to
1970. He was a pacificist who spent time in jail for
his views during WWI. Amazingly, his Nobel
Prize came in literature, not peace or the sciences.
His philosophy was based in suing mathematics
to ground logic, as well as in metaphysics. None
of this means much to mean, I just like his quote.
The philosopher Bertrand Russell said, “In all affairs it's a healthy thing now and then to hang a question mark on the things you have long taken for granted.” The study of biology is such a good example of this idea. We should always be questioning the ideas that we have come to accept, and science should welcome efforts to overturn longstanding concepts.

This is why I don’t mind efforts to disprove evolution - IF they are carried out in a scientific manner – it’s the only way science can truly work. Unfortunately, many efforts are not carried out scientifically. They're similar to that old cartoon, where one scientist is charting the mechanism of a process on the board, and one of his steps is, “and then a miracle happens.”

On the other hand, when a radical change comes to us in a scientific manner, then we should accept it is another possible explanation; now we have multiple explanations to try and disprove. We're never be done testing a hypothesis.

And that brings us to the reptiles; the testudinata (turtles and tortoises, from Latin = movable armored shed), the squamata (snakes and lizards, from Latin = having scales), and the crocodilians (from Greek = lizard). Of these, the turtles and tortoises seem to be the oldest, appearing first more than 220 million years ago. Lizards were next, and snakes were said to have developed from lizards, about 100-120 million years ago. This is harder to follow in the fossil record because snake skeletons are delicate and do not fossilize well.

The snakes get all the publicity; I think it's the big fangs and the fact that they can swallow things bigger than their heads! Children can list the most venomous snakes, boys especially like to taunt young girls with writhing snakes; they even make movies about snakes on airliners!

But did you know that only about 20-30% of all known snake species are known to be venomous? Of the non-venomous majority, only the boas and the anacondas seem to get any attention. Their immediate ancestors, the lizards seem to get even less.

Gila monsters are black with pink, orange, or yellow
patterns. As such, it is one of the few animals with
pink pigmentation. Their tails are plump, and can store
fat for times when food is rare and over the winter.
There is a type II diabetes drug made from a protein
isolated from gila monster saliva. It is sometimes
called “lizard spit.”
Until recently only two lizards were known to be venomous, both calling North America home. The gila monster (Helderma suspectum) has a bite that, though usually not fatal to humans, can cause paralysis, convulsions, and difficulty breathing. It doesn’t have fangs that inject the venom into the flesh, but its saliva contains the toxin and he will literally chew on you, driving the poison into the wound. Toxic slobber, that’s all we need.

His cousin, the Mexican bearded lizard (Heloderma horridum) lives a little south of the gila, and he is considerably bigger, reaching up to a meter in length. The bearded lizard’s venom delivery is similar to the gila’s, with modified salivary glands in the lower jaw producing venom that flows with the saliva.

I said that recent events have changed our ideas about venomous lizards. Brian Fry at the University of Melbourne has published a series of papers since the early 2000’s that have expanded our understanding of venom evolution. Dr. Fry reported in early 2006 that beyond the heloderma lizards, the monitors and the iguanas also use venom in defense or food acquisition. One of the monitor lizards is the Komodo dragon (Varanus komodoensis), the largest living lizard on Earth.

Well, that’s all anyone remembered - the Komodo is venomous?! This really caught the public’s attention. It had been assumed that it was the vile state of the Komodo’s bacteria-infested mouth that was toxic to its prey, the bite transferred horrible organisms to the wound, which replicated, let loose with some bizarre toxin, and eventually killed the prey. Whole documentaries were made about how patient the Komodo was, following the prey around for days until it succumbed to one infection or another.

This is a computer rendition of the MRI of a Komodo
head. The dragon has six venom glands on each side of
the lower jaw, shown here as alternating red and pink.
The yellow glands produce the mucus that
gives the Komodo its famous drool.
Entire research programs were set up to discern how the Komodo could live with such terrible microorganisms in its saliva. Why didn’t the Komodos die too? Teams searched for new antibiotics in the saliva and blood of the Komodo. Now it turns out that the Komodo had a different strategy all along! Its venom is strong enough on its own to kill the prey animals. Fry’s follow-up 2009 paper showed the venom was the principal killing weapon of the Komodo; it's an anticoagulant and induces shock, so the victim bleeds out and its organs fail.

Two items were lost from Fry’s 2006 paper. One, the iguanas have venomous members as well. Iguanas live in North America as well as in Madagascar, Fiji, and Tonga. They have many representative species, several of which are common household pets. It turns out that many of them have venom of a mild potency, but strong enough to hold some predators at bay. The monitors and the iguanas have something else in common, the way the produce and deliver venom.

In both groups, the venom glands are located in both the upper and lower jaws, and the ducts that deliver the venom to the saliva are located between the teeth and at several locations in the mouth. This is different from the snakes that have venom glands only in the upper jaw and deliver venom through a single duct, and the gila monster and bearded lizard that have venom glands only in the lower jaw.

The differences between gila/bearded lizard venom delivery and that of snakes had led to the idea that these two systems developed independently of one another. But following the trail backwards through evolution, Fry showed that venom production started in the lizards, about 200 million years ago. The snakes that evolved from some lizards already had the mechanisms on board to produce venom – it came from a single common ancestor that was around before even T. rex!

The pink iguana is native to the Galapagos Islands.
It isn’t known whether it is a venomous iguana.
They were only discovered a few years ago and
were thought to be a variant of the common land
iguana, but genetic tests showed it was a
novel species on its own.
The two-jawed venom glands of the monitors and iguanas are ancestors to the single-jaw venom glands of the heloderma and the snakes, some evolved to house it only in the upper (snakes) and some evolved to house the glands only in the lower (gila monster and bearded lizard).

This was a radical change for those who studied the evolution of the reptiles. Like I said, one should never consider a theory to be absolutely proven. The questions then is - if lizards started the reptile venom idea, why are so few venomous now? Well, maybe they are venomous. We only seem to care if humans are threatened by them, so we haven’t even looked for venom or venom glands in most lizards. Fry estimates that perhaps more than 100 lizard species are venomous, and that perhaps all of them still contain the genes to produce venom.

Two of Fry’s co-authors jumped on the idea of all lizards being venomous, and expanded it to the idea that perhaps all snakes might have been venomous at one time. As such, they suggested a new categorization of all these animals into one clade (evolutionary group with a single common ancestor), called the Toxicofera clade. This idea became know as the Toxicofera Hypothesis… makes sense, I guess.

This is a cladogram that shows the hypothetical Toxicofera, or
venom clade. The emergence of venom glands occurred
about 200 million years ago, so all descendents of this animal
have the potential to be venomous. This includes ALL the
snakes (serpentes), the gila and Mexican (heloderms), the
Komodo and monitors (Varanid), and the iguanas. The one
in the middle that looks like a snake is actually the group of
worm lizards, their ancestor diverged before the
emergence of venom glands.
The Toxicofera (“those who bear toxins”) hypothesis is universally accepted, but it is gaining influence and has started discussions about the relative ages of venomous animals and who evolved from whom. It has also sparked investigations into seemingly non-venomous snakes and lizards, looking for remnants of venom glands and venoms. It is completely possible that some snakes and lizards are completely non-venomous, but they may still harbor the genes for venoms or some vestigial (regressed and unused) venom glands.

But don’t misunderstand the hypothesis. It applies only to reptiles, the Toxicofera family would not include venomous mollusks, like the blue-ringed octopus, or biting ants, like the fire ant, or the stinging bees and wasps. There are certainly instances where venom and venom delivery systems evolved independently. The toxicofera hypothesis applies only to the reptiles.

How about the turtles and tortoises? They are reptiles too; could they be venomous? Well…. no, at least not yet. These reptiles diverged from a common ancestor more than 200 million years ago, before the computer programs estimate the first reptile venom appeared. However, there is one sort of exception. The three-toed box turtle of North America isn’t venomous, but it can be poisonous.

In the wild, three-toed box turtles eat just about anything – insects, small dead animals, vegetation, worms, and fungi; it's the fungi that make the difference. They have a taste for the Death Cap Mushroom (Amanita phalloides). This fungus is related to the toadstools that some people use for their hallucinogenic properties, but the Death Angel is aptly named.

On the left is the three-toed box turtle, native to North
America. It is an exceptional in that it is the only turtle
that can completely close itself in it’s shell. You can
see the hard flap under its chest that will close the opening
when it ducks its head inside. On the right is the
Amanita phalloides death cap. It is very common and
very deadly. Its toxin is alpha-amantin, an RNA
polymerase II inhibitor, that causes complete
liver breakdown.
If the box turtle has eaten some of these mushrooms, the toxin will be in its tissues for some time. If you happen to like turtle stew, you could end up meeting the Angel of Death. Grilling up a wild box turtle was supposed to have made several boys very sick in the 1950’s, but I haven’t found another instance in the literature of this occurring any more recently. The question is - why don’t the mushrooms kill the turtle?

This is something we can investigate next week, along with the difference between just having a poison in your tissues because of something you ate or actively storing it for your own use. Along the way, we will meet a salamander with a lethal rib cage. Yeah- you read correctly, ribs that can kill.


Fry, B., Vidal, N., Norman, J., Vonk, F., Scheib, H., Ramjan, S., Kuruppu, S., Fung, K., Blair Hedges, S., Richardson, M., Hodgson, W., Ignjatovic, V., Summerhayes, R., & Kochva, E. (2005). Early evolution of the venom system in lizards and snakes Nature, 439 (7076), 584-588 DOI: 10.1038/nature04328  

Fry, B., Wroe, S., Teeuwisse, W., van Osch, M., Moreno, K., Ingle, J., McHenry, C., Ferrara, T., Clausen, P., Scheib, H., Winter, K., Greisman, L., Roelants, K., van der Weerd, L., Clemente, C., Giannakis, E., Hodgson, W., Luz, S., Martelli, P., Krishnasamy, K., Kochva, E., Kwok, H., Scanlon, D., Karas, J., Citron, D., Goldstein, E., Mcnaughtan, J., & Norman, J. (2009). A central role for venom in predation by Varanus komodoensis (Komodo Dragon) and the extinct giant Varanus (Megalania) priscus Proceedings of the National Academy of Sciences, 106 (22), 8969-8974 DOI: 10.1073/pnas.0810883106  

Fry, B., Winter, K., Norman, J., Roelants, K., Nabuurs, R., van Osch, M., Teeuwisse, W., van der Weerd, L., Mcnaughtan, J., Kwok, H., Scheib, H., Greisman, L., Kochva, E., Miller, L., Gao, F., Karas, J., Scanlon, D., Lin, F., Kuruppu, S., Shaw, C., Wong, L., & Hodgson, W. (2010). Functional and Structural Diversification of the Anguimorpha Lizard Venom System Molecular & Cellular Proteomics, 9 (11), 2369-2390 DOI: 10.1074/mcp.M110.001370
For more information, see:

Venomous lizards –
http://animal-world.com/encyclo/reptiles/lizards_venomous/Venomous.php
http://scienceblogs.com/notrocketscience/2009/05/18/venomous-komodo-dragons-kill-prey-with-wound-and-poison-tact/
http://animals.about.com/b/2009/05/19/komodo-dragons-pack-a-venomous-bite.htm
http://www.abc.net.au/science/articles/2005/11/17/1506321.htm
http://www.mapoflife.org/topics/topic_388_Venom-and-venom-fangs-in-snakes-lizards-and-synapsids/
http://www.pbs.org/wnet/nature/lessons/righteous-reptiles/lesson-activities/4683/
http://www.reptilesalive.com/teachers/teacherlessons.html
www.reptilesalive.com/teachers/teacherlessons.html
http://www.animallaw.info/articles/biusreptile.htm

Toxicofera hypothesis –
http://www.sworch.com/modules.php?name=Reptiles-MM&page=Toxicofera.html
http://www.askabiologist.org.uk/answers/viewtopic.php?id=4981
http://www.pueblozoo.org/articles/Monitorlizard.htm
http://www.hermanaresist.com/category/relationships/
http://lib.bioinfo.pl/paper:22446061
http://www.sworch.com/modules.php?name=Reptiles-MM&page=Toxicofera.html

Three-toed box turtle –
http://exoticpets.about.com/od/boxturtles/p/threetoeboxt.htm
http://www.tortoisetrust.org/care/ctriungis.html
http://mdc.mo.gov/discover-nature/field-guide/three-toed-box-turtle
http://users.ccewb.net/lonerock/turtles/NaturalHistory.htm
http://www.bio.davidson.edu/people/midorcas/research/Contribute/box%20turtle/boxinfo.htm
http://nationalzoo.si.edu/animals/reptilesamphibians/facts/factsheets/easternboxturtle.cfm