Roberta Score was riding a snowmobile over the snowy landscape on December 27, 1984 when she spotted a dark spot on the bluish ice. The spot was a rock, the first of many she would pick up in the Allen Hills of Antarctica that year, all of them meteorites.
Roberta Score was paid to search for rocks and noticed nothing special about this one, named 84001, 84 denoting the year, 001 showing that it was the first rock. At the end of the season, 84001 and the year’s other rocks were packed up and sent to storage. Once there, like most of the rocks collected, like most specimens of any kind, it was ignored.
The rock was in no hurry. It had been traveling a long time, roughly 4.5 billion years since the day it formed on Mars. Its biggest trip occurred when a meteorite knocked the rock free of Mars 16 million years ago. It then took the back roads of space until thirteen thousand years ago when it landed in Antarctica, where Robbie Score found it when she came over the hill.
Things changed for the rock in 1994, ten years after it was first discovered, four billion years after it formed. A geochemist, David (“Duck”) Mittlefehldt, analyzed a tiny chip of the rock in a study of asteroid fragments. The piece was identical to rocks known to be from Mars. That the rock was from Mars was quickly uncontested.1 Also uncontested was that the rock was older than any individual stone on Earth.* It was oldest of the few known Martian meteorites. Yet it was an unremarkable kind of thing, a medium-sized, green-black rock. If it landed on your lawn, you might not notice. Other meteorites, possibly even older meteorites, have probably landed in gardens, then been cursed and tossed into the weeds.
Scientists split the Martian rock into pieces and then studied those pieces for evidence of life. What happened next happened quickly and with increasing secrecy. Chris Romanek, a NASA scientist, discovered structures in the rock that resembled, in form, bacteria. Another NASA scientist, Kathie Thomas-Keprta, discovered magnetic crystals in the rock, similar to the crystals formed by some bacteria. The physicist Richard Zare from Stanford University found polycyclic aromatic hydrocarbons (PAH), molecules in the rock that can also be found in living cells. Other evidence of life in the rock accumulated. Then David McKay, the NASA geologist in charge of the project, found forms that were segmented and that looked to him even more like fossil bacteria.2
McKay, the most senior of the secret collaborators, and colleagues wrote a paper and published it in the journal Science.3 Even before the publication, there was an announcement. David Goldin, NASA’s administrator, arranged the kind of press conference that Woese could only dream of. President Clinton looked into the cameras and announced:
Today, rock 84001 speaks to us across all those billions of years and millions of miles. It speaks of the possibility of life. If this discovery is confirmed, it will surely be one of the most stunning insights into our universe that science has ever uncovered. Its implications are as far-reaching and awe-inspiring as can be imagined. Even as it promises answers to some of our oldest questions it poses still others even more fundamental. We will continue to listen closely to what it has to say, as we continue the search for answers and for knowledge that is as old as humanity itself but essential to our people’s future.
A few months after the press conference, Sagan, by then already very sick with the blood cancer that would kill him, appeared on the television program Nightline. He had waited a lifetime for evidence of living creatures on Mars. On Nightline, Ted Koppel asked him about McKay’s find. Sagan responded simply that, “If the results are verified, it is a turning point in human history.” Verification would take time, in the end more time than Sagan had. Koppel, perhaps seeing the signs of decline in Sagan, asked if he did not have some personal thoughts for the audience, thoughts, Koppel may have implied, about life or death. Sagan, without stopping to brood or ponder, offered simply, “These are our true circumstances. We live on a tiny ball of rock and dust, in a cosmos vast beyond our imagining.” Not long after that interview, at two minutes after midnight on December 20, 1996, Carl Sagan died. Life had not yet been conclusively found on Mars, but Sagan’s great light had gone dark.
Daniel Goldin, the NASA administrator who organized the announcement of the Mars meteorite, would write upon Sagan’s death, “Everything we learn about Mars will carry some of the seed of Carl Sagan’s dreams.” Sagan’s shadow looms over all research about Mars, the story of 84001 included. If the Martian rock held life, it would be vindication of his life’s work, the germination of that seed.
The Martian rock had withstood billions of years of wear. Now it had to withstand the scientific method. Each of the pieces of evidence would be finely dissected. The machinery of skepticism went to work, looking for holes and fundamental weaknesses. Scientists like to find weaknesses in each others’ discoveries almost as much, maybe more, than they like to make their own discoveries. Within two years, hundreds of responses to the paper by McKay and colleagues were published, and those responses had raised doubts. In 1997, one hundred and twenty planetary scientists were asked what the probability was that the microbes in rock 84001 were real. The median response (half of the answers higher, half lower) was 20 percent, which is to say an eighty percent chance of being nothing at all.4
There were several problems. The rock looked as though it had withstood temperatures too high for life. The magnetite minerals, first interpreted as shaped as though tempered by life, proved more variable than initially expected.* The focus, though, was on one uncontestable reality of the rock: the putative Martian microbes were very, very, small. The forms were not just smaller than any known microbial life, they were many times smaller. The forms were, depending on exactly how one measured, about fifty nanometers across, fifty billionths of a meter, one twentieth the diameter and just one one-hundredth of one percent of the volume of, for example, the bacterium E. coli.5 Two thousand of the Martian bacteria would have fit into a human blood cell. They were just too small to be uncritically accepted as evidence of life.
The polycyclic aromatic hydrocarbons—so similar to those found in life on Earth—were suggestive but not conclusive on their own (they are also found independent of life), and the same was true for the particular minerals (carbonates) that had seemed worthy of interest. Many wondered whether the putative fossil forms, too small to be any known microbe, were big enough to be life at all.
There were at least two historical models for what results when one scientist, but not others, sees life where it is not expected. Leeuwenhoek discovered microbes. Lowell discovered an imaginary world. In either case, history predicted that the scientists who discovered the microbes in the rock and became their advocates would stick to their guns for the rest of their careers, looking more and more deeply into that original rock and then also at other Martian rocks. McKay, in particular, obeyed the prediction, to the scorn of those critical of his find, and the praise of those who supported him. Whether he was a genius or something less flattering would take time to decide.
When one scales down to the size of the fossil microbes in the Martian rock, interpretation becomes art. One is up against the limits of microscopy. Forms that are obvious at bigger scales become open to interpretation, vulnerable to preconceived ideas about what is and is not possible. Even scanning electron microscopy is not high enough in magnification to see such small shapes well. More elaborate methods of microscopic visualization, methods few were capable of doing, needed to be used.
There was, however, a precedent for tiny life. In 1985, the Finnish biologist Olavi Kajander was studying mammal cells in culture in California.6 Kajander was interested in the sterility of blood and cell cultures. He was working as a postdoc in a laboratory where sterility testing was a big part of the day-to-day work. He found that despite using sterile techniques, his cultured cells would very often die, and he had no idea why. He became curious about the possibility that human and animal blood could contain something that would pass through the existing tests, something, perhaps, too small to be detected. He approached the question simply at first.
As Kajander finished his postdoctoral fellowship and went back to his native Finland, he continued to wonder what else might live in blood, what might have been killing the cells he was studying. The problem seemed amenable to resolution, and chance was about to reward his prepared mind. The opportunity awarded to Kajander was a total lack of funding for research. As Kajander tells the story, had he had a big grant, he would have studied one of the well-behaved and standard questions. He would have added detail to some well-known story. He would have worked fast, because funding and publication demand speed, and he wouldn’t have had time to dally on strange and obscure questions. Kajander wouldn’t have time to look into what he calls “artefacts and exceptions.” Instead, he was fortunate. No one had funded him. He had time, so he did the simplest thing he could think of; he looked at the cells.7
Kajander began with the free serum samples from cows, serum that companies had provided him as a marketing ploy. In those samples, he began to see something—both the haze of discovery and, he would soon believe, an entirely new kind of life. He did nothing more than look at the samples, samples from serum grown in a variety of different mediums. It was, initially, just that simple, but what he found was surprising. Inside the pure cultures there was a kind of fog in the liquid and a white spot at the bottom of the beaker, on one side. It seemed to Kajander, almost immediately, to be something new. He wondered whether what he had found was life, life too small to be called life by the definitions of the day, life small enough to pass through the filters meant to exclude life, but life all the same.
He spent the next four years repeating his experiments, publishing nothing, and then finally writing a small grant. Inside the dying cells were tiny, apparently living, forms. As he continued to work, and to verify his initial results, he imagined the significance of what he had found. Here was a new kind of life, a kind of life that might be common, ubiquitous even. Here was, at the very least, the turning point in Olavi Kajander’s life. He had, like his predecessors in discovery, begun with an observation, something he literally saw. That observation did not fit with what scientists “knew” to be true, but as Kajander would himself later say, “it is the artefacts that matter, the artefacts that are important, the artefacts that we need to explain.”8
In 1990, Kajander had reached a point of no return. He had applied for a grant, so others now knew of his discovery, but he did not get the grant and therefore had relatively few choices. To stake his claim on this intellectual territory, as a kind of stopgap until funding arrived, he applied for a patent on nanobacteria on May 8, a patent for the “culture and detection method for sterile-filterable autonomously replicating biological particles.” In doing so, he named them, giving them the Linnaean genus name, Nanobacterium. It took two years, but the patent made clear that, at least according to the patent office, Kajander had isolated life, life that was distinct from viruses because it reproduced on its own and distinct from known bacteria because it was, frankly, so incredibly small. That patent would hold off the hounds, he imagined, until he could do more, but he had underestimated the hounds.
They came quickly, gnashing teeth. The controversy would follow immediately and would revolve around one of the same problems faced by McKay and his team when they announced their findings on the Martian rock. The forms were too small, by many scientists’ measures, to be alive. At fifty nanometers, they were nearly identical in size to the Martian microbes.
No one knows how small the smallest life is. A few years ago, most microbiologists who had thought to wonder about the question might have suggested something around eight hundred nanometers. New finds have pushed that limit. In 2002, a new microbe was discovered in a submarine hydrothermal system off the coast of Iceland. The form, named Nanoarchaeum equitans by its discoverer, Karl Stetter, was an archaean of an entirely new phylum. It measured just four hundred nanometers in width, half the size of the smallest microbe known at that time. It had been undetectable prior to the study in which it was found, invisible to the common methods being used.9
For microbiologists, this phylum was a big discovery.10 As University of Halifax biologist Ford Doolittle put it, it was “as worthy of our notice as any coelacanth or other macroscopic living fossil.” The Nanoarchaeum appear to live parasitically on a larger archaean called Ignicoccus. Nanoarchaeum are small in part because they take advantage of the chemical machinery of other cells. Still, the smallest form of life was getting even smaller.* In addition, as two microbiologists commentating on the discovery noted, “there is a lingering suspicion—or hope, depending on your disposition—that there might be even weirder organisms hiding out there.” A recent study suggests that most of archaea species might be essentially invisible to existing methods for finding microbes.11 Some microbiologists still believe we might yet find something as new as a fourth domain of life, to go along with the eukaryotes, archaea, and bacteria. It is a romantic notion, to be sure, but one that is not at its surface ludicrous.
Yet, even as the smallest life shrinks and the possibility of another domain is talked about by serious scientists, most microbiologists still believe that there are lower physiological, or simply physical, limits to the possible size of a living cell. If you take an E. coli bacteria cell and pare it down to the bare minimum of necessary genes and parts, the DNA, RNA, and ribosomes alone are big enough to require a cell at least two hundred nanometers.12 At a conference to determine the minimum viable size for living cells, the palaeontologist Andrew Knoll of Harvard University offered that he thought “everyone pretty much agreed that…nothing much smaller than 200nm is likely to be viable.” Life smaller than two hundred nanometers was “not compatible with life as we know it.”13† One hundred nanometers, the biggest dimension of both Kajander’s nanobacteria and McKay’s Martian microbes‡ remained, to most, too small. In the same article in which the find of Nanoarchaeum was lauded, the authors felt compelled to note, “Unfortunately, the name invites comparison with nanobacteria, a much more problematic group. It is unclear that nanobacteria, which are often much smaller entities…could house the minimal machinery necessary for life….”14
Despite these criticisms, Kajander’s initial discovery seemed to open up new avenues of research. Kajander and his collaborator, Neva Ciftcioglu, found more nanobacteria wherever they looked. The nanobacteria were widespread, found throughout the human body, and throughout nearly every body, particularly where things have gone wrong: in ovarian cancer, in kidney stones, in cases of heart disease. It had long been known that one could find calcifying particles associated with many diseases.15 Kajander, and soon others, began to think it was plausible that those particles were nanobacteria. Nanobacteria have now been associated with many diseases, a sort of who’s who of bodily bad news: rheumatoid arthritis, cholecystolithiasis, coinfections with HIV, various cancers, Alzheimer’s disease, prostatitis, and even lowly periodontal disease. The work on links between nanobacteria and kidney stones in particular has been the subject of research by scientists at the Mayo Clinic and elsewhere. Frederic Coe, a kidney stone specialist at the University of Chicago, called the links between nanobacteria and kidney disease “one of the most intriguing and fascinating additions [to the study of kidney stones] I can imagine.”16
Kajander and Ciftcioglu were becoming, through their nanobacterial research, a new pair of Leeuwenhoeks, or maybe Leeuwenhoek and Pasteur. They had, through observation, broadened dramatically the scope for life. No one had ever imagined life to be so small. Like their predecessors, Kajander, Ciftcioglu, and their team found something that had been invisible.
As before, with the discovery of the past lives of mitochondria and chloroplasts, strong support for Kajander and Ciftcioglu’s discovery would require them to find RNA (or DNA, for that matter) and to use Woese’s method to understand the position of whatever genes they might find on the tree of life. It was not enough to see the nanobacteria: the nanobacteria had to have parts—nucleotides, enzymes, and proteins—the levers and pulleys of life that could be compared to those of other life-forms. Then it happened. In 1998, Kajander and Ciftcioglu isolated RNA from one of their samples. The RNA indicated the nanobacteria were a kind of proteobacteria and so suggested the possibility of billions of proteobacteria, all smaller than it had been predicted bacteria could be, living in our bodies, passing through the filters used to clean blood, moving around us invisibly. Their team had discovered the smallest life on Earth, and it was everywhere.
The isolation of RNA from nanobacteria was, for Kajander and Ciftcioglu, a moment of wonder. They had made one of the most important discoveries in a hundred years. It was too bad, then, that few other scientists agreed with their results. Shortly after the publication showing RNA in the nanobacteria, another team showed that the RNA Kajander had found most closely resembled that of a bacterium common on laboratory equipment.17 The results appeared contaminated and so Kajander and Ciftcioglu went back to the drawing board. So far, that is the end of the story of nanobacterial RNA, and we wait. Even Kajander now suspects that nanobacteria do not reproduce using nucleic acids. For him, the big story of what the nanobacteria are is being obscured by the academic debate about whether or not they ought officially to be counted as life.
Kajander, his colleague Ciftcioglu, and their now separate (but still collaborating) groups are among just a handful of scientists who continue to work on nanobacteria.* Almost all publications on nanobacteria (many, but not all, in obscure journals) have been published by Kajander, Ciftcioglu, or their colleagues. Like discoverers before them, like Margulis, Woese, or even Leeuwenhoek, they are their own strongest advocates. The papers are rejected, but the scientists persist.
One of the reasons there has not been more study of the forms Kajander and Ciftcioglu have isolated is that until they are better documented, it is risky to study them. No granting agency wants to fund work on an organism that may or may not exist (though this is precisely what Lynn Margulis did).* In fact, any funding that did exist has since dried up. At least to Kajander’s knowledge, no grants have been awarded for work on nanobacteria in eight years.18
The other problem is more complex. Kajander and Ciftcioglu are not employed by universities. Kajander was a professor at Kiobe University in his native Finland, but another Finnish professor accused Kajander of academic fraud, arguing, in essence, that since nanobacteria do not exist, the evidence for them can only be fraudulent. The case was eventually dismissed, but the damage was done. Kajander lost both his job and his grants. He was again without money for his research, but this time he resorted to more unusual means: he started a business with the plan of developing treatments for nanobacteria, which he had begun to believe were not only small, but problematic.
Among academics, economic success—or even the possibility of economic success—is always suspect, particularly so when that success hinges on a particular, hard-to-verify scientific result. Kajander and Ciftcioglu were until very recently scientific advisors for a company now called Nanobac Life Sciences, dedicated to developing techniques for ridding people of their nanobacteria. Together they started the ancestor of Nanobac, which was later bought out and converted into a publicly traded company on the New York Stock Exchange. The more common and problematic nanobacteria are, the more money Nanobac stands to make. Therefore, Kajander and Ciftcioglu had a financial incentive to find nanobacteria in more and more places. The more they found, the more evidence that people will need treatment to counter their effects. Kajander and Ciftcioglu were perceived to have the temptation toward bias, in the same way that Lowell looking to Mars saw cities where he had always hoped they would be.
Everyone, including Kajander and Ciftcioglu’s staunchest critics, agree with a few things. They have discovered something. That thing can reproduce and it can be transferred in liquid, for example, blood samples, from one individual to another. Most also agree that the nanobacteria do not respond to antibiotics, and survive boiling temperatures. Some scientists suspect that nanobacteria are real, but are a kind of crystal with lifelike attributes (including the ability to cause disease), but without meeting the criteria of life.* Others are less generous, but everyone agrees that Kajander and Ciftcioglu’s nanobacteria theory is bold. Pasquale Urbano and Francesco Urbano of the Italian Ministry of University and Research said of the nanobacteria theory that it “is no less revolutionary than the famous germ theory of disease, which was put on solid ground by the efforts of Pasteur and Koch, or the one on contagium vivum fluidum, which heralded the birth of virology.” What the Urbanos disagreed with was whether or not the theory was right.19 A great deal is at stake in these seemingly arcane debates in obscure journals and meetings. Kajander now claims that one in two people on Earth die from diseases caused by nanobacteria. If he is right, he is very, very right. But if he is wrong, he is very wrong.
Leeuwenhoek, Erwin, Woese, and others were at the fringes of their respective fields, the frontiers, to be generous. The same might be said to be true, as the Urbanos point out in their article, of Galileo. Those discoverers were vindicated, but their ideas started out at the very margins of believability. If we are to look for the next big discoveries, discoveries of entire biological realms, the place to look may not be the big, well-funded labs of well-respected scientists. The place to look may be to the very fringes of science. That is exactly where one would find Olavi Kajander and Neva Ciftcioglu, but are these two too marginal and wild? Another recent quote by Urbano and Urbano seems to sum up the sentiment among some or even many biologists: “Galileo Galilei was vindicated by history, Kajander and Ciftcioglu so far only by the Web: in a Google search for nanobacteria [we] get 90,500 hits with Google, and 198,000 with Yahoo.”
Kajander and Ciftcioglu seem to meet the first requirement of big discoverers by being outside the mainstream of science. Time will tell if they meet the second requirement: being right.
Meanwhile, there has been a new study of nanobacteria by a group of French scientists led by Didier Raoult at the Unité des Rickettsies, Centre National de la Recherche Scientifique. The study, which involves scientists from seven different institutions, was released in February 2008 with little fanfare.20 The group thinks they have discovered, once and for all, what nanobacteria are and they have accordingly given them a new name: nanons.
The group began by repeating some of Kajander’s experiments and they, for the most part, found that the nanons did what Kajander said they would. The nanons grew in fetal calf serum. They formed a cloudy film at the bottom of the culture flask. The nanons could also be transmitted from serum with nanons to serum without nanons through fine-grain filters. When the nanons were exposed to enzymes that destroy RNA and DNA, they were not altered. Nor were they affected by antibiotics. All of this was known from Kajander’s work, but to have it verified independently was significant.
Next, Raoult’s group began new tests. They showed that the nanons could be destroyed by UV or gamma radiation or by contact with acidic conditions or compounds that bind calcium. They showed that nanons were, when magnified with electron microscopy, simple, dense, concentric rings of circles, like a bull’s eye, surrounded by a “loose halo.” They were larger than those Kajander had first detected (431 nanometers on average). Slowly, some details were emerging. Raoult’s group then repeated tests to see if the nanons had pathogenic effects. They did. When amoebae were exposed to the nanons, they died, as did fetal calf cells. When the nanons were introduced into mice, the mice had an immune response.
Putting everything together, Raoult and colleagues had begun to develop a theory that might explain these disparate observations. Perhaps the particles contained a kind of protein, a fetuin, in a mineral complex. Fetuin is a blood serum protein that is particularly common in fetal serum. Its role is not well understood, but it is thought to inhibit calcification. If the nanons contained fetuin, they should stain when antibodies to fetuin were applied to them. They did this and so they were, it appeared, either pure fetuin or some other substance that was dense with fetuin. If the nanons contain or are fetuin, much remains to be resolved. It is not clear how the fetuin replicates itself. It is not clear how the fetuin complexes kill cells and cause pathogenic effects. Suddenly, though, Raoult and colleagues have done two things. They have, in a body of very rigorous work, substantiated a suite of Kajander’s claims about his basic observations of the nanobacteria (or nanons, if you prefer). They have found, like other studies, that the nanobacteria can have negative effects on cells. The paper is careful to say that the nanobacteria are not living, but the question of whether they are life depends more on definitions of life than it does on the findings at hand. Whatever they are, they are, as Kajander first thought, new.
Among the mysteries that remain, and there seem to be a growing number, is the question of why Kajander initially found RNA with, on, or in the nanobacteria. As for Kajander, he has a theory that he thinks explains everything. He said in a phone interview that “[he] had now solved the problem of exactly what nanobacteria are,” and that he “now knew conclusively,” but would wait until he had funding to publish his results. So we must wait, too.
The Martian microbes had been approximately the size of nanobacteria, and so a quiet idea inevitably began to float around that perhaps the nanobacteria and the Martian microbes were one and the same. Together, perhaps they were another origin of life, of Martian life that had been around us for years but just missed by standard techniques. The idea persists. It has even been suggested that nanobacteria represent an early, Martian life from which we are all descended. Another study, this one by two British scientists, J. T. Wickramasinghe and N. C. Wickramasinghe, suggests that the optical refractive index of our galaxy (how much light is bounced around by particles in space) is what might be expected if there were lots of particles in the galaxy about the size of nanobacteria. In other words, nanobacteria are so dense they alter how we see our galaxy.21 The pair has also gone on to argue that nanobacteria in clouds spread disease around the world. When the going gets tough, the tough get, well, a little crazy.
As for Kajander and Ciftcioglu, the discoverers of nanobacteria on Earth, and McKay’s team, the discoverers of nanobacteria in a Martian rock, their careers are now inseparable. Ciftcioglu and McKay worked together for a while before Ciftcioglu returned to Turkey. Whatever the fates of nanobacteria and life in the Martian rock, they will be forever linked. David McKay has now found what he contends is more evidence of life on Mars, this time from the Nakhla meteorite. McKay now has pictures of the meteorite that show tiny pits, pits McKay has interpreted as similar to those produced by some microbes as they bore into rocks. Again, the criticism will begin and with time we will know what the consensus is among scientists. With more time, we may even know whether that consensus is right.