Microbiological analysis of Mammoth Mountain: the experience of investigative journalism

About mysterious microbes unearthed in the permafrost of Yakutia, they have been writing all sorts of nonsense for a long time. But either someone wrote something that even the authors of the idea of prolonging life with the help of frostbitten antediluvian bacteria ran out of patience, or vice versa - British Siberian scientists drove another wave of self-promotion. Is it both, and their left hand doesn't know what their right hand is doing? Or maybe the group split into opposing factions… I started the investigation with a note in RIA-Novosti, to which a lady teacher came and told an interview.

With a lot of oddities and inconsistencies. But, as it turned out in the course of the investigation, the beginning is a bad thing, then more.

Scientists urge the media not to rush with the "discovery" of a cure for aging MOSCOW, June 28 – RIA Novosti.

Research work with paleobacteria found in permafrost on the territory of Yakutia has just begun and it is impossible to say that they are supposedly able to slow down aging and increase life expectancy, Marina Zenkova, head of the laboratory of Nucleic Acid Biochemistry at the Institute of Chemical Biology and Fundamental Medicine of the Siberian Branch of the Russian Academy of Sciences, where experiments are underway, told RIA Novosti.

"The experiments were done only on mice, preliminary results were obtained. We are seeing a slight increase in life expectancy and an overall improvement in the condition of the animal. But these are not yet published data, these are still preliminary experiments, but what journalists turn it into – the hair stands on end," the agency interlocutor said. On Monday, some media reported that this research group allegedly found microorganisms that prolong human life by 30-40 years.

That they get up – I agree, but, first of all, this black Monday happened for the first time a year and a half ago: Google issues microorganisms of Mammoth Mountain in the first line: January 20, 2009... Life-prolonging bacteria found on Mammoth Mountain in Yakutia… And then – a bunch of articles, including interviews with the head of the research group, Doctor of gerontology... no, for some reason, geological and mineralogical sciences Prof. Brushkov's run-out in the weekly Yakutsk Evening. With a bright name "Yakut mammoths will give a cure for old age". However, nothing gets up from the text, the professor mostly gets rid of the annoying correspondent with hints, although promising, but a specific figure is given in the subtitle – who will answer for this bazaar, a professor or a journalist?

And at the same time, another lady, her colleague and co-author, ran around the editorial offices and promised immortality to everyone and for nothing, and let no one leave offended. But we'll get to it a little later.According to Zenkova, scientists are really studying an ancient bacterium of the genus Bacillus cereus, found in permafrost near Mammoth Mountain in Yakutia in layers dating back to about two million years ago.

However, so far, with the help of experiments on mice, scientists have only managed to make sure that this bacterium is not dangerous, as well as to obtain some data on its immunostimulating properties.

"This bacterium is a kind of immune system stimulator. Maybe someday we will figure out the mechanism, and it really will be a kind of dietary supplement that will allow to slow down aging in people who lead a healthy lifestyle," the agency interlocutor said.

The deeper into the permafrost, the thicker the misunderstandings. In the reference article (but about it later), the age of the layer from which this microbe was extracted is not specified, only they write about permafrost that it is 3.5 million years old (not 2!), and cereus is not called a cure for old age, but they write like this: "... apparently, it is a new species. The greatest species similarity of the isolated bacillus was noted with Bacillus simplex, B. Macroides...".
And about cereus in the article on the Rusmedserver it is written, in particular, this:
"B.cereus produces several toxins, including necrotizing enterotoxin, emetic toxin, phospholipase C, proteases, hemolysins and enterotoxins... infections caused by B.cereus are divided into 6 broad groups:
• local infections, especially in the area of burns, traumatic and postoperative wounds and eye infections;
• bacteremia and septicemia
• CNS infections, including meningitis, abscesses and bypass-related infections
• respiratory infections
• endocarditis and pericarditis
• food toxicoinfections characterized by toxin-induced vomiting and diarrhea. Are there any volunteers here for clinical trials of a potential geroprotector?

Or do you think cereus is a typo? No! A couple of days before, on June 26, in another, no less scientific source – "KP" – the above-promised other lady from the same company said even cooler:

Novosibirsk scientists are testing the "longevity bacterium"...The bacterium was discovered several years ago and belongs to the well-known genus Bacillus.
Its closest relative is Bacillus cereus, which many of us take as a probiotic "Bactisubtil," says Nadezhda Mironova, a senior researcher at the Institute of Chemical Biology and Fundamental Medicine SB RAS, Candidate of Biological Sciences.

As it is not difficult to guess from the name or, for the unintelligent, to Google in 2 clicks, Bacti subtil is made from B.subtilis, a harmless representative of the intestinal microflora. But let's return to the analysis of the first interview.She (again, M. Zenkova, at RIAN) stressed that before studying the effect of bacteria on life expectancy, it is necessary to determine in what concentration they should be administered to experimental animals.

It took a year to determine the toxicity, to find ways to work safely with this bacterium.

Found, found concentration! And the mice were injected intraperitoneally with doses from chutosh to equine, and no one had peritonitis, but on the contrary, they lived about 20% longer than the control group. After the expansion of a single small dose at the age, in terms of human, pre-retirement.
Hey, hey, Aubrey de Grey!
Send the prize of Methuselah to Tyu-men!

Be patient a little more – we'll soon get to the sensations....

Zenkova explained... "The mouse lives for about a year. Such an experiment should be carried out for two years on the same mouse, and it should live this time under careful supervision. We will only be preparing for this experiment, these are very expensive and difficult experiments," the agency interlocutor concluded.

I won't even explain what exactly is wrong in this paragraph: a solid delirium.An ancient bacterium, which presumably can slow down aging, was discovered in permafrost on Mammoth Mountain (Verkhoyano-Chukotka region, Yakutia) by a group of permafrost researchers from Tyumen State Oil and Gas University with the help of microbiologists from the Institute of Chemical Biology and Fundamental Medicine SB RAS.

This bacterium has a unique ability to self-preservation, since DNA molecules have been preserved for only hundreds of thousands of years.

Hundreds of thousands and the discrepancy of this "only" with the above two million will be attributed to noise in the microphone. But it is very, very strange that the head of the laboratory, who will only be preparing for difficult experiments, is not in the courses that they have been held for a long time. Only if their sensational results were at least a little bit like science more convincing, the article would have been published not on the back of the website of the Tyumen Scientific Center SB RAS, but
In kind, I give a tooth!
Another representative of the same teachers is in the courses. True, the figures in the secret article and in her interview do not converge a little, and the method of introduction, too, and many other things, but these are trifles. A finger from the ceiling and not such mistakes can be obtained. Here's what she told a few days later, and not to our special correspondents, but to the foreign ones (Found in nature's freezer, the secret of living to 140):

The bacterium Bacillus F may allow a person to live up to 140 years NEWSru-medicine, July 1, 2010

A bacterium trapped in permafrost conditions may allow a person to live up to 140 years, scientists say. They have already managed to double the lifespan of mice and flies by isolating Bacillus F from ice, writes The Daily Mail… Nadezhda Mironova, a representative of the Institute of Chemical Biology in Novosibirsk, where the research was conducted, states: "We had a control group of mice with an average life expectancy of 589 days. After intramuscular administration of the bacteria to another group of rodents, their life expectancy increased by about 308 days. On average, they lived 906 days"... The next stage of research will be conducting clinical trials, the essence of which will be reduced to an attempt to create a real medicine from bacteria.

589 + 308 = somewhere 897, and 589 X 2 = somehow 1178, but let's not be petty. And do not miss the underlined one: it will come in handy if you have the patience to finish reading to the appropriate place in the summary of the article. But to start research on living people on the basis of such excretory experiments… I don't believe it! (C) The full text of the academic article (ahtung!

a lot of science-like letters!) with all the typos, including the last sentence of the conclusions: "... allows us to consider the research of relic microorganisms in gerontology promising.about the prevalence of grooming activity." lies here. And below is quoted and commented on the most delicious.

RELICT MICROORGANISMS OF CRYOLITHOZONES AS POSSIBLE OBJECTS OF GERONTOLOGY At a time when the study of the mechanisms of aging is still an urgent fundamental problem, the study of cells capable of surviving for many millennia may well be of interest to gerontology…

...Evidence of the viability of microorganisms in permafrost appeared in the nineteenth century. In 1979, bacteria, fungi, diatoms and other microorganisms were discovered at the Vostok Antarctic Station... their age is about 500 thousand years… ...Calculations show that even small fragments of DNA (100-500 nucleotides) can last no more than 10,000 years in normal climates and up to a maximum of 100,000 years in cold areas… Thus, it seems unclear how bacteria survive in the thousand-year permafrost. The ability of relict microorganisms to remain viable for a long time implies the existence of mechanisms that prevent the accumulation of damage. It seemed to us expedient to study the influence of these microorganisms on higher organisms, especially since the immune reactions of the latter may be of interest… Did you notice the powerful logic in the fragment I underlined?

Did you understand from which oak tree the idea fell on the heads of Siberian scientists not to study the survival mechanisms of frostbitten bacteria (and first make sure again that this is not contamination), but to see how they will affect higher organisms? I didn't get it either. Unlike the author of the above-mentioned note in Komsomolsko Pravdo: "... This bacterium was discovered several years ago in permafrost in Yakutia, on Mammoth Mountain.

Have you realized? Because it ends with a "Y"! Marx's teaching is omnipotent because it is true! (C)MATERIALS AND METHODS OF ISOLATION OF MICROORGANISMS RESULTS OF ISOLATION, STUDY OF GROWTH AND IDENTIFICATION OF MICROORGANISMS

...the sediments from which the samples were taken were obviously in a permafrost state... the age of the permafrost on Mammoth Mountain can reach 3.5 million years. In addition to the described outcrop, samples ... were taken from younger re-vein ice of the Yakutia ice complex, from the walls of the dungeon of the P.I.Melnikov Permafrost Institute in Yakutsk, as well as from underground ice in the Fox tunnel and at the gold mine near Fairbanks in Alaska. Samples of frozen rocks were taken with the greatest possible precautions for field conditions…

Nothing significant has been removed here. The age of the samples is unknown; which of them sprouted in Petri dishes is also a mystery; there is no certainty that the bacteria on the mountain, and even more so in the basement, tunnel and mine, are antediluvian, and not modern. But the enthusiastic teachers are sure: these are very, very ancient microbes!Ribosomal DNA of microbial culture was extracted using Fast DNA kit for soil…

Fragments of 16S rRNA were amplified by polymerase chain reaction... amplicons were cloned using pCR2.1 vector... plasmid DNA was sequenced on ABI PRISM 3100 Genetic Analyzer... Amplified products 27F-1492R were sequenced in both directions with primers 27F, 357F (5'-CTACGGGAGGCAGCAG -3'), 520R... The greatest species similarity of the isolated bacillus It was noted with Bacillus simplex, B. macroides, homology with 16S rRNA of which is 96-97%.

Well done, boys! I just remember the old movie "Surly River", the faithful kunak of Proshka Ibrahim-ogly and his phrase: "Harashe is bursting, you bastard"! Then, however, he writes much worse. THE NATURE OF LONG-TERM VIABILITY OF MICROORGANISMS

Fig. 3. Results of nucleotide stability studies. The half-life of cytosine at temperatures of 0 - -10 degrees Celsius for about 300 years is about 350 yearsSam, vosem – what's the difference: and the schedule is very tricky, however, and it ends at 20 degrees…

But what is the point of this chapter at all? They wrote at the very beginning "science is unknown"! And that's right, with such a half-life time (this is how they usually write about biomolecules, so as not to be confused with physics), no millions or even tens of thousands of years can last. And this is for individual nucleotides – what can we say about the whole bacterial chromosome! And indeed, all this blah-blah-blah led to the conclusion "this mystery is great":Thus, the long-term existence of living microorganisms is difficult to explain by the slowing down of vital activity during suspended animation.

Apparently, it is due to the presence of unique repair mechanisms that support cellular structures.

TESTING OF BACILLUS CULTURE ON HIGHER ORGANISMS

Testing on Drosophila melanogaster fliesFor the experiment were selected…

They were placed ... an experimental variant, when the first 5 days of flies were kept on a medium with the addition of yeast, then 1 day on a medium with a bacillus ... there was a decrease in fertility in relation to the control by 5 times ... there was an excess of the proportion of surviving flies from the 24th to the 42nd day of the experiment compared with the control (fig. 4).

 
Rice. 4. The effect of Bacillus sp. culture, strain 3M, on the lifespan of Drosophila melanogaster

In middle and old age, the mortality of flies in the experiment was slightly lower than in the control, in senile – slightly higher. The maximum life expectancy from the microbe has decreased, which, according to the authors, the piano does not play. The terrible inflammatory disease "does not stand"The authors considered the fivefold decrease in fertility a trifle. They did not consider the average life expectancy for experience and control, the confidence interval, the median fashion. And we made two logical conclusions:
– Dietary supplement with microbe prolongs the life of flies;
– and now we will prick it in the abdomen of the mice.

Testing on laboratory mice...In the first series of experiments, the effect of culture doses on the parameters of the immune system of young animals (age 3-4 months) was studied.

Two groups of animals were used in the control, one of which was intact, and the second group was injected with a saline solution. Bacterial culture was administered to animals once intraperitoneally 5,000 (1)... and 50,000,000 (5) microbial bodies (mt) per animal.

Culture of bacilli in a small dose (5000 mt.) stimulate, and in medium doses (500,000 and 5000000 mt.) – suppress the phagocytic activity of splenic macrophages ... in almost all doses increases the activity of humoral immunity, and a dose of 5000 mt. promotes an increase in the functional activity of both cellular and humoral immunity…

In this regard, a dose of 5,000 m.t. was chosen to study the effect of culture on life expectancy.... In the second series of experiments, the effect of bacterial culture on the physiological and behavioral reactions of "elderly" mice (age 17 months) was evaluated, while the culture was administered once intraperitoneally at a dose of 5,000 m.t. The control group was represented by animals of the same age... The minimum life span of mice from the control group was 589 days, and the maximum was 833 days. The minimum life span of mice from the experimental group was 836 days, and the maximum – 897 (Fig. 6).

Rice. 6. The effect of Bacillus sp. culture, strain 3M with parenteral administration of 5000 cells on the life expectancy of laboratory mice aged 17 months

I looked at the chart for a long time, like a goat on a poster, until I realized that it should start not from 0, but from 17 months. By the way, it still needs to erase the last two round marks on it, otherwise it turns out that the difference in maximum life expectancy is 1 month, not 2. The authors calculated the average duration later, preparing for an interview with the Daily Mail, and during the journey she was able to grow by exactly forty days, I remind you, 906 days. Or 1178? Although a simple estimate (836+897)/2 gives 866.5.
The rest of the statistics are missing, and what is there to count, on two groups of 15 individuals, of which five in the control (!) for some reason died much earlier than the deadline (see graph)? Even with such an impressive difference in maximum life expectancy (either 7.7%, if according to the text, or 3-4%, if according to the schedule – there are still about 30 days to 833), the planks of confidence intervals will climb far, far behind each other, and the error of the average will be the size of the average itself. And tricky methods for small samples are from the evil one. Finally, I will analyze one more oddity, chronological.

The expedition in which this bacillus was unearthed took place in August 2008.

The date of publication of this very scientific article is not specified, but right under it in the heading “science-idiots ideas” there is a transcript of the lecture delivered on 31.01.2008, and a little higher – the report "And there will be a Tyumen schastye" from October 2009.
It turns out that the comrades of the teachers managed to isolate their microbe of dubious longevity, multiply, identify, feed them flies, beat some mice, then others, make a bunch more gestures, wait a year until the last of the mice of the experimental group dies – and put it all in no more than the same year?
Something doesn't add up here. Either the bacillus was dug up 3-4 years before, or the article was written in advance, for future results. But whatever it was, you can call this article scientific only by lying very much to your soul. Or without understanding anything in any science, even geological and mineralogical.
Wherever you poke – solid absurdities. It is enough that in the first experiment, on young mice, there was no third control group that was injected not with saline, but with a culture of some harmless modern bacteria. By the way, a priori it seems to me that this could affect the immune system in the same way as frostbitten miracle bacilli. And not for the rest of my life, but for a while. Are there immunologists in the hall? Help me figure it out! And in the second experiment (let's forget about the strange schedule), the control group was not even injected with saline. Ftopku! I.e., the design of the experiments does not comply with the rules known to any excellent student over 15 years old. Against the background of such blunders, statements like this do not cause any confidence:

The body weight of the animals of the experimental group was higher than that of the animals of the control group 2 months after the introduction of the culture. Muscle strength in experimental animals increased (about 60%) relative to peers from the control group.

And from the conclusions made by the authors, I agree only with the latter, and then – with the exception of what I crossed out:
3. The reduction in mortality of Drosophila melanogaster flies in individual experiments, as well as laboratory mice in combination with stimulation of the immune system and improvement of the physical condition of the latter allows us to consider the research of relic microorganisms in gerontology promising.about the prevalence of grooming activity.A Healthy Skeptic

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05.07.2010