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Visulalizing 2 nanograms....i.e. 0.000000002 grams

19K views 81 replies 17 participants last post by  JRG13 
#1 ·
2 nanograms is how much neonicotinoid poison it takes to kill a honey bee. Hard to visualize isn't it. So... maybe the follwing would be a better perspective.

Assume a healthy hive has 60,000 bees. In Texas, there are about 80,000 hives, so thats 4.8 billion bees (60,000*80,000 = 4,800,000,000).

4.8 billion x 2 nanograms = 9.6 grams (4,800,000,000 * 0.000000002 = 9.6)....and that just about fills 2 tablespoons...

Get it....2 tablespoons of a neonicotinoid could kill every bee in the state of Texas...
 
#2 ·
You could also see it from this prospective:
it kills bees at molecular level.
1 mole contains 6*10^23 molecules, Imidacloprid, MW=256, 256g/liter==> 1M imidacloprid; 2 ng of imidacloprid ==> 1.28*10^-12M === > 7*10^(23-12) ==> 7*10^11 -->> 70000000000 molecules per bee.

Honey bee has 960000 neurons, so EACH neuron in bee's tiny brain would be "poisoned" by 70000 molecules of Imidacloprid.

Note - I am horrible at arithmetic, please excuse me if I made any mistake in simple multiplication/division. I would gladly correct my calculation if somebody will find an error. BUT scale of "disaster" is right!
 
#3 ·
2 nanograms is how much neonicotinoid poison it takes to kill a honey bee. Hard to visualize isn't it. So... maybe the follwing would be a better perspective.
...
You do realize that neonicotinoids are a class of chemical compounds and that there is no leathal dose for a class of chemicals?

Strike 1

If you are talking about the most used neonicotinoid, you do realize that you are incorrect in your figures?
From Wiki:
Like most insecticides, imidacloprid is highly toxic to bees, with a contact acute LD50 = 0.078 μg a.i./bee and an acute oral LD50 = 0.0039 μg a.i./bee.
Strike 2

You do realize that ther are 1,000 nanograms in a µg? Strike 3... You're outta here! Already.

Oh and you didnt take specific gravity into account for your tablespoon calculation.

Strike 4

Get it? (obviously not)
 
#4 ·
From Wiki:
Like most insecticides, imidacloprid is highly toxic to bees, with a contact acute LD50 = 0.078 μg a.i./bee and an acute oral LD50 = 0.0039 μg a.i./bee.
Well, 0.078 μg x 1000=78 ng and 0.0039 μg x 1000 = 3.9 ng, which is 2x more than in original post, but it could not change the picture - the bottom line is that it is VERY small amount! I do not like your aggressive tone - you sounded rude and uneducated... :( If you so knowledgeable - just do all right calculations, determine the specific gravity of imidacloprid powder (I want to see how you could do it) and give us YOUR estimate and we'll see how it is different from original post. Be friendly...
 
#7 ·
The calculations are not hard, and it is interesting (and maybe useful?) to be able to visualize these things.

There are many estimates of LD50 for imidacloprid on honey bees, but let's take a published value of 0.008 µg (8 ng) per bee. (http://www.cdpr.ca.gov/docs/emon/pubs/fatememo/Imidclprdfate2.pdf)

A worker bee weighs about 100 mg. Thus the toxicity of imidacloprid to honey bees is 8 ng/100 mg = 0.08 mg/kg, if we use the same units as LD50 for mammals. This is about 600 times more toxic than nicotine, 80 times more toxic than cyanide, 20 times less toxic than ricin, and 8000 times less toxic than botulinum, the most human-toxic compound known. Not quite snake venom, but at the same time no compound with this level of toxicity to humans would be allowed to be freely dispersed into the environment.

The LD50 of imidacloprid to rats (the best proxy for humans) has been estimated at 450 mg/kg. Thus the chemical is over 5000 times more toxic to bees than it is to us. Of course that level of specificity is desirable in an insecticide, but when your livestock happen to be insects that is a problem.

It's not easy for me to visualize 80,000 hives, so I'll stick with one. One hive with 60,000 bees. Killing half the bees in a hive would thus require 480,000 ng, or 0.48 mg of imidacloprid. Seed corn is treated at rates of 0.25 to 1.25 mg per kernel. (http://www.extension.iastate.edu/CropNews/2012/0406hodgson.htm) At the higher rate, one kernel contains enough imidacloprid to kill half the bees in two full strength hives. At the lower rate it would take two kernels to kill half the bees in one hive.

This analysis of course ignores the fact that bees are affected at dosages much lower than the LD50, and it makes the assumption that the chemical would somehow be distributed equally to all bees in the hive, which is false. But still, corn is planted at the rate of 35,000 seeds per acre, and one or two seeds contain enough imidacloprid to kill a whole hive. Corn planting is a dusty endeavor, and not all of that imidacloprid stays on the seed. That would be enough to scare me if I kept bees in corn country, or at least convince me to move my hives well away from fields during planting time.

I don't know that we need to ban imidacloprid and its related neonic cousins, but we need to realize that from a bee's perspective we are dispensing a chemical 80 times more toxic than cyanide across wide expanses of ag lands adjacent to hives. We definitely need rules (isolation distances and times) to improve coexistence.

Mark
 
#8 · (Edited)
A worker bee weighs about 100 mg. Thus the toxicity of imidacloprid to honey bees is 8 ng/100 mg = 0.08 mg/kg, if we use the same units as LD50 for mammals. This is about 600 times more toxic than nicotine, 80 times more toxic than cyanide, 20 times less toxic than ricin, and 8000 times less toxic than botulinum, the most human-toxic compound known. Not quite snake venom, but at the same time no compound with this level of toxicity to humans would be allowed to be freely dispersed into the environment.
Using rats as a proxy for humans is an acceptable practice. Comparing mammals to insects in toxicology analysis is unacceptable and downright ludicrous. (In case you didnt know, they use rat proxy for humans because they can't test humans. They regularly do tests any bees, rats, and fat head minnows). Why not save the contorted argument and look up the LD50 for nicotine, cyanide, ricin, bot toxin for honeybees directly?

but we need to realize that from a bee's perspective we are dispensing a chemical 80 times more toxic than cyanide across wide expanses of ag lands adjacent to hives.
Uh, you might want to check and see if cyanide is actually toxic to honeybees before you throw statements like that out there.


But anyway you missed the whole question…

How many tablespoons?
:lpf:
 
#9 ·
I have to agree with Nabber - you say it only would take 2 tablespoons full of really toxic stuff to kill all of the bees in Texas and he proves you you are so so wrong - it would take at least 4 tablespoons - probably heaping tablespoons at that - to kill all of the bees in Texas. You completely lose all credibility and Nabber proves it. Good Job Nabber.

Clearly if it takes ANY MORE than 2 tablespoons of this stuff then it just isn't a problem. Unless they use more than 4 heaping tablespoons in the entire state of Texas - which they don't. Right?

Or was it teaspoons? My head hurts.
 
#10 ·
My head hurts.
As it should because you have no concept of exposure pathway(s). One simply does not round up the entire bee population of Texas and make them eat 2 tablespoons of imidacloprid. How exactly would that be accomplished?

Just as if I collected all the cyanide in the world, rounded up the entire world population, and made them eat it (party in Jonestown anybody?), it would kill all of the people in the world. Big deal.
 
#12 · (Edited)
Comparing mammals to insects in toxicology analysis is unacceptable and downright ludicrous.
All I did was to convert the LD50 for bees (8 ng/bee) into per weight units (0.08 mg per kilogram of bee weight) using the known weight of an average worker bee. Thus IF imidacloprid were as toxic to mammals as it is to bees, it would have an LD50 of 0.08 mg/kg, which can be compared to other mammal-specific LD50's. Clearly it isn't that toxic to mammals (as I clearly pointed out), but I'm using the comparison to make the valid point that from a bee's perspective imidacloprid is a very toxic chemical, right up there with ricin and cyanide.

And if you must know, imidacloprid has a density of 1.543 g/cm3
(http://pmep.cce.cornell.edu/profiles/extoxnet/haloxyfop-methylparathion/imidacloprid-ext.html)

Thus the 0.48 mg required to kill 50% of the bees in one strong 60,000-bee hive would occupy a volume of 0.00031 cm3. One tablespoon is 14.79 cm3, so one full tablespoon of imidacloprid could deliver an LD50 dose to 47,700 of our 60,000-bee hives.

If you want to be really picky that is for crystalline imidacloprid. Powder has a lower packing density, so the volume would be slightly different.

I'm not trying to fear-monger and say that neonics are responsible for all of our bee problems. Just pointing out that there is no chemical with anywhere near this level of toxicity to humans that is in widespread use.

One kilogram of cyanide (rat LD50: 6.4 mg/kg) could theoretically kill 1040 75-kilogram humans.
One kilogram of imidacloprid (bee LD50: 8 ng/bee = 0.08 mg/bee kg) could theoretically kill 83,000 75-kilogram bee-masses. That's about 63 billion bees, or one million strong hives.

I realize that neonics are a hot political topic these days, with lots of misinformation on both sides. But we can't lose sight of the fact that these chemicals are exceptionally toxic to bees, and that any guidelines that minimize bee exposure can only help beekeepers.
 
#14 ·
Thus IF imidacloprid were as toxic to mammals as it is to bees, it would have an LD50 of 0.08 mg/kg, which can be compared to other mammal-specific LD50's.
Then why on earth are you making that comparison? You are not even going across species of mammals (rats to mammals), you are crossing from arthropods to mammals.
 
#20 ·
As a side note, the more complicated and tortuous the pathway the less valid.
I wasn't aware exactly how incredibly toxic neonics are to bees until I did these calculations today. With this level of toxicity, complicated/tortuous pathways must be considered.

If 1/35,000th of the amount of neonics in an acre planted with treated corn seed is enough to kill 60,000 bees, then we need to consider not only direct routes of exposure, but also more complicated ones (e.g. bees picking up imidacloprid dust while in the air, planting dust settling on adjacent blooms, bees collecting pollen from corn plants grown from treated seed, etc.). If neonics were 10- or 100-fold less toxic, these pathways wouldn't be of concern.

It's hard to accidentally ingest enough cyanide to die, but if you had a bottle of ricin all it would take is one breath of wind to send a few particles into the air. And that's unfortunately the level of toxicity we have with neonics and bees.
 
#24 ·
If 1/35,000th of the amount of neonics in an acre planted with treated corn seed is enough to kill 60,000 bees, then we need to consider not only direct routes of exposure, but also more complicated ones (e.g. bees picking up imidacloprid dust while in the air, planting dust settling on adjacent blooms, bees collecting pollen from corn plants grown from treated seed, etc.). If neonics were 10- or 100-fold less toxic, these pathways wouldn't be of concern. .
I suppose that I could go on to explain how to quantify the probability of completing a pathway and how to assess the risk of any particular pathway being completed, but I am growing weary of this tedious exchange. Yaaaawn. Good night and sweet dreams.
 
#21 ·
I don't know nabber. The op said that it would take a tiny amount to kill a whole bunch of bees, then you twisted it up by saying that if it was the particular chemical that YOU chose then it would take a lot more of it, and you would have to make them eat it first anyway, so strike three. Nabber wins. I would ask if I got any of that wrong, but I already re read it, and I didn't.

And yes I understand that toxicity was measured by mixing the poison with syrup and feeding it to the bees, and farmers don't do that, so it must be harmless if you follow the directions.

I also understand that these chemicals are insecticides, and bees are insects. Some of them act systemically, by making the entire plant more or less an insecticide - thus killing pests that try to eat those plants. Whether it is true or not it makes sense that bees which gather pollen or nectar from such a plant could be carrying tiny doses of poison back to the colony. And true or not it also makes sense that when that hive ingests that poison while also under seasonal stress, that it could cause winter colony collapse.

Neonicitinoids may not have a thing to do with the high percent of colony loss we have seen this year. I certainly can not prove that they do, but you can't prove that they don't. And no one who is in any position to do anything about it cares one bit about what either one of us thinks.
 
#22 ·
Nabber wins.
You got that right holmes.

I would ask if I got any of that wrong, but I already re read it, and I didn't.
Not sure how to respond to that because it isnt a coherent sentence, but I think that my response is that you you did indeed, get it wrong.

You asked and I tried to help. I can't do any more than that.
 
#32 ·
And here is where the paralysis of analysis becomes a tactic to avoid accountability.

We cannot afford to cloud over for a problem that needs to be addressed. Nabber86; your desire to raise awareness of inappropriate calculations, although cloaked is cynicism and contempt, is quite impressive. I also admire your desire to preserve the fidelity of the scientific method and the purity of the “Hard Science.” We all could stand to embrace more precision so the facts are truly facts, and not just sensationalized speculations.

Fact – These chemicals exist, are used where bees frequent, and are highly toxic when ingested by bees.
Fact – Bees eat, and bees feed other bees.
Question – Do the toxins exist on food sources that bees eat . . . most likely.
Question – Do bees eat items on which the toxins are present . . . most likely.
Question – Are bees ingesting the toxins . . . most likely.
Fact – A very, very, very small measure of these toxins can kill a bee when ingested.
Fact – The toxins are made to kill insects
Fact – Bees are insects.
Question – How much toxin is UNHEALTHY for bees . . . not certain
Question – How much toxin is actually ingested by bees . . . not certain

However, no matter what variables exist in the study, measure, and calculation of this enormous problem, at least people are thinking deeply and willing to search out the etiology. The set of causes, or manner of causation, of this condition deserves some sensationalism in order to awaken the minds of humanity. Just like you are raising the awareness of others to calculation errors, raising the awareness of people to this very serious pandemic is also worthy; more than worthy of theater. Drama draws attention. Yes, this drama must be based on accurate fact. However, instead of plowing down others in what appears to be apathetic arrogance, how about utilizing your prowess to assist the goal at hand; to stop CCD and preserve the honey bee? Isn’t that what this is really about? In short, we can either be part of the problem, or part of the solution. Choose wisely.
 
#33 ·
BW: While I agree with much of what you are saying, it's important to note that aside from some confirmed cases of bee losses from planter dust (a problem that has supposedly been solved) bees have no direct exposure to these seed coatings. Yet all these threads seem to start with the underlying assumption that neonics are the cause of massive bee deaths despite the fact that the evidence proving this is virtually nonexistent. These are all still theories. The fact that these chemicals are so wide spread in their use yet many beekeepers still have great success in building strong bees says to me that bee health is a very, very complex issue.
 
#34 ·
You are right Jim - it is clearly a complex issue. Mites, 2 kinds of nosema, all manner of chemicals, pollution, SHB, viruses that we haven't even heard of, monoculture ag, heat, drought, and dearth - most, if not all - of these things are probably involved in high colony loss numbers. It's too complex to say that any one thing is the cause, or that any one thing is not a factor. But we will probably quibble about the details from now on - cause that's what people do.

If the internet has taught us anything it is that when someone says "my thing isn't what's at fault in this broad issue it's everything else that is the problem" to take that with a grain of salt at least. The same goes for those who blame big problems on one person, or one group of people.

Most likely all of these things are going to continue to affect bees and bee keepers for the foreseeable future. Our options are very limited in what we can do to adapt - but most likely adapt we will.

I hope so anyway.
 
#35 ·
Wow this was an interesting thread, hats off to nabber for picking on the math and not rolling over and ignoring it!!!
AND the others who are pointing out errors...

We also want to point out neonics being leathal, but that is not even close to the only "poisen" spread on a huge basis daily. from spraying mosquitoes for west nile to anhydrous for corn.....we as humans do spread chemicals. BUT we look closely at the impacts.. spraying mosquitoes doesn't effect bees because we take due care. same for corn seeds... due care and tons of math have been done to study what really matters The UPTAKE of the chem in question..... Doing silly math to point out it takes less than a lb (being generous) to kill all the bees is a moot point...... getting it to them is another matter. As mention... how much Rican would it take to kill all the humans?? small amount also.. but worthless math as its not going to happen.....
 
#36 ·
Well....looks like I accomplished exactly what I started out to do....get some serious discussion going. So I remembered the toxicity quantity wrong by a factor of 2 or 4...or even 10.....

What if I'm really off by a factor of 1000 and it doesn't take 2 tablespoons of a neonic to kill all the bees in Texas...but 20 POUNDS.

Boils down to a really simple question....Do I really want that around my bees? Uh......NO.

Lets see if I can really fire up Nabber and mention the soil latency issue...which I have seen stated as 1,300 plus days depending on the soil type.

I've recently seen a figure quoted for degradation in the soil of 60% per year.....i.e....of what ever percentage makes it to the soil, 60% is still there a year later....36% two years later...22% three years later...etc.

Assuming the same field and dose rate per year, the second year you get another 100% dose...so now you have 160% soil level (100% year 2 +60% of year 1)

Year three would be 100% (year 3) +60% (year 2) + 36% (year 3) = 196%

Year four would be 100% (year 4)+ 60% (year 3) +36% (year 2) +22% (year 1) = 218%.

Looks like bioaccumlulation to me. OK so lets say its not 60% degradation per year, but 30%...still doesn't matter....the only way it wouldn't bioaccumulate is if it degrades completely before the year's next application....

Ok Nabber...you're on...
 
#38 ·
Boils down to a really simple question....Do I really want that around my bees? Uh......NO.

Lets see if I can really fire up Nabber and mention the soil latency issue...which I have seen stated as 1,300 plus days depending on the soil type.

I've recently seen a figure quoted for degradation in the soil of 60% per year.....i.e....of what ever percentage makes it to the soil, 60% is still there a year later....36% two years later...22% three years later...etc.

Assuming the same field and dose rate per year, the second year you get another 100% dose...so now you have 160% soil level (100% year 2 +60% of year 1)

Year three would be 100% (year 3) +60% (year 2) + 36% (year 3) = 196%

Year four would be 100% (year 4)+ 60% (year 3) +36% (year 2) +22% (year 1) = 218%.

Looks like bioaccumlulation to me. OK so lets say its not 60% degradation per year, but 30%...still doesn't matter....the only way it wouldn't bioaccumulate is if it degrades completely before the year's next application....
So, keep your bees off other peoples property. Do you think that just because you have bees you can tell other people how to use their land?

Most farmers tend to rotate the crops grown in a field. So, a field may not have the same pesticides applied to it year after year.

If you are successful in getting neonics banned what is going to replace them? Do you think the carbamates, organophosphates, pyrethrins, etc. of the past are a better option?

Exaggeration to get a discussion started is planting false premisses. The uninformed will take the exaggerations as fact and multiply them. It also detracts from the bigger picture. While neonics have the potential for killing bees no one has linked them to supposed CCD colony deaths. There are other threats to bee colony that them get diminished because people believe the neonics HAS to be the cause.

Tom
 
#39 ·
soil latency is funny issue. In buried soil that has no biotic effects it takes up to 1,300 days to degrade but in soil exposed to sun and water neonic based chemicals photo-degrade and hydro-degrade with a half life measured in hours (not years). That's one of the little factoids that always seems to get forgotten in the standard sensational articles on neonic pesticides.

So the exposed pesticide degrades rapidly and the entombed pesticide stays resident for long periods of time. Of course I fail to understand how the neonic pesticide buried in the soil will get to the bees in ways where photo-degrading and hydro-degrading wil not occur. all this and the observation observed in the Randy Oliver article linked previously that bees don't much like neonic flavored nectar or pollen and will avoid eating it (when given a choice).
 
#40 ·
soil latency is funny issue.....
Yes. Sun would work only if it is direct sun - top of the sand's grain would be exposed to sun, the bottom - no. Also, water. I would imagine that water will be where there is no direct sun due to evaporation. So ether sun or water... it means that decomposition on the sun, will be much less that you think. For photochemical decomposition usually water is required. Also, grown plants will block sun on the soil - again, less decomposition. Farmers I guess, turn or mix soil at the end of the season in preparation for spring - it will bury some chemicals... Bees will bring some chemicals inside the hive - it is dark inside and water, perhaps, is limited... beehive itself is a perfect storage for pesticides etc.

All these arguments that nasty chemical(s) regularly spread in environment may be harmless for numerous reasons is just showing how little many people at beesource do care about environment they live... :(

I really like Bee Whispers sentence:
"In short, we can either be part of the problem, or part of the solution. Choose wisely. "
 
#41 ·
cerezha,
we had a whole thread on this a while back that you also partook in that I don't want to rehash but to summarize:

sun + water = half life of hours
sun and no water = half live of days

most soil surface has water in it every day except deserts (rain, dew, ambient moisture in the humus, etc).
again the soil gets buried wont photodegrade, but then again it's not directly accessable to the bees (not that any neonic on the surface is that accessible anyway)
Farmers tilling the soil? Depends on if they are going with a 'till' or 'no till' approach. I'm about 20 years removed from my parents farm but I thought most farmers were going with 'no till' these days to minimize erosion (hence the need to eradicate all the existing weeds with herbacide)

top of the grain exposed / bottom of the grain not exposed - not completely accurate, but ground isn't completely flat, nor is all of its components completely opaque. (and most farmers don't farm in sand).

Sergey,
the details you are pointing out seem awfully minor and I don't see how they really add up to a significant issue.
 
#43 ·
...Sergey,
the details you are pointing out seem awfully minor and I don't see how they really add up to a significant issue.
Exactly! But this approach (distraction from the main point by criticizing minor details or imperfections) is widely used here to diminish the important substance - I used this approach just to show how primitive it is and how it just distracts from important issue and does not create any useful conclusion. Look at the title of the thread: "Visulalizing 2 nanograms....i.e. 0.000000002 grams." Visualizing! Nothing else! To me the essence of this particular thread was how little is needed to kill so many bees. It is educational for me because I never did such estimates and I am thankful to jeb532 for such interesting look on known thing.
 
#42 ·
Beekeeping is unique among agricultural pursuits in that beekeepers "farm" the surrounding 15,000 acres for honey, but almost never do they own that land. Since bees are generally beneficial this practice is accepted. Unfortunately then, the beekeeper must also accept the loss incurred due to bee-harmful activities within their foraging area, and there isn't much legal recourse. Nor should there be, I would argue, particularly if the landowners have no real need of the bees (as is true with corn, pasture, and all crops outside of their pollination window).

This sort of wild-farming, unrestricted by property boundaries, has no modern parallels. Think of it as if deer were owned by keepers, who became angry whenever one was killed on a road or inadvertently poisoned.

What cannot be solved with rules, then, can sometimes be solved with incentives. Certainly beekeepers have incentive to keep bees away from pesticide-heavy areas, and that is something I would always consider when choosing a home or an outyard. The next step would be to create financial incentives (bee easements?) for landowners in particular areas to limit use of bee-toxic products.

Think of it as a land trust for bees. The goal would be to create foraging circles of roughly three-mile radius in which all properties carried a bee-safe certification, rather like the salmon-safe and other labels currently in use. Just an idea...
 
#47 ·
Sergey, honestly the subject title is a sensational topic name. It's a great technique to propagandize ...

Here we go again, another distraction from you on the science of all this
From other posts, I gather that serg has a scientific background. I am completely baffled as to why anyone that claims to be a scientist continues to discuss the toxicity of neonics based on non-science (nonsense).
 
#51 ·
Sergey,
the best way to get students to understand a concept is to use an example that exaggerate the concept's effects to make it stand out visually (ie. 'using liquid nitrogen and a balloon to demonstrate the thermal dynamics of air' , 'using a solid glass barrier to demonstrate the principle of greenhouse gases', etc). In that capacity I think you do well.

That said, what is your technique that you would personally use to visualize a measurement or proportion or or an equation where exaggeration will not be an effective tool.
Examples would be:

'the poison is in the dosage'
'how much neonic can a bee consume safely and at what rate can metabolize it'?
 
#52 ·
Sergey,
the best way to get students to understand a concept is to use an example that exaggerate the concept's effects to make it stand out visually...
It is American way. It did not work in real science... Normally, a math or chemical formula/equation is to visualize the concept in many cases. For instance, how you could visualize 5th dimension? Just math formula. Or how you could imagine the principle of electron's uncertainly (electron may be in two points at the same time)?

'the poison is in the dosage'
'how much neonic can a bee consume safely and at what rate can metabolize it'?
It is interesting topic and we could discuss it off-line on better scientific level. But, one need to keep in mind that evil of neonics is their mechanism of action - they irreversibly binds to receptor - thus, they do not metabolize and just accumulate. In such situation, it is very difficult to determine the "dosage" because even sublethal amount may cause a problem.

The visualization for that would be - neurons could not talk to each-other it means that muscles are not coordinated, memory lost ... is it enough?
 
#54 ·
The original post was aimed at developing a visualization, and I think that this has been accomplished: neonicitinoids have the potential to be very toxic to bees in what seem to be very small amounts.

What's happening here is that three separate discussions are taking place. One conversation is aimed at honing the visualization through application of better math, a second attempts to illustrate how the visualization is based on a variety of assumptions and the last attempts to demonstrate how the visualization would not be accepted as peer-reviewed scientific theory. Everyone is correct.
 
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