Metal-Eating Bacteria Could Have Left their “Fingerprints” on Mars, Proving it Once Hosted Life

Today, there are multiple lines of evidence that indicate that during the Noachian period (ca. 4.1 to 3.7 billion years ago), microorganisms could have existed on the surface of Mars. These include evidence of past water flows, rivers and lakebeds, as well as atmospheric models that indicate that Mars once had a denser atmosphere. All of this adds up to Mars having once been a warmer and wetter place than it is today.

However, to date, no evidence has been found that life ever existed on Mars. As a result, scientists have been trying to determine how and where they should look for signs of past life. According to a new study by a team of European researchers, extreme lifeforms that are capable of metabolizing metals could have existed on Mars in the past. The “fingerprints” of their existence could be found by looking at samples of Mars’ red sands.

For the sake of their study, which recently appeared in the scientific journal Frontiers of Microbiology, the team created a “Mars Farm” to see how a form of extreme bacteria might fare in an ancient Martian environment. This environment was characterized by a comparatively thin atmosphere composed of mainly of carbon dioxide, as well as simulated samples of Martian regolith.

Metallosphaera sedula grown on synthetic Martian Regolith. The microbes are specifically stained by Fluorescence-In-Situ-Hybridization (FISH). Credit: Tetyana Milojevic

They then introduced a strain of bacteria known as Metallosphaera sedula, which thrives in hot, acidic environments. In fact, the bacteria’s optimal conditions are those where temperatures reach 347.1 K (74 °C; 165 °F)  and pH levels are 2.0 (between lemon juice and vinegar). Such bacteria are classified as chemolithotrophs, which means that they are able to metabolize inogranic metals – like iron, sulfur and even uranium.

These stains of bacteria were then added to the samples of regolith that were designed to mimic conditions in different locations and historical periods on Mars. First, there was sample MRS07/22, which consisted of a highly-porous type of rock that is rich in silicates and iron compounds. This sample simulated the kinds of sediments found on the surface of Mars.

Then there was P-MRS, a sample that was rich in hydrated minerals, and the sulfate-rich S-MRS sample, which mimic Martian regolith that was created under acidic conditions. Lastly, there was the sample of JSC 1A, which was largely composed of the volcanic rock known as palagonite. With these samples, the team was able to see exactly how the presence of extreme bacteria would leave biosignatures that could be found today.

As Tetyana Milojevic – an Elise Richter Fellow with the Extremophiles Group at the University of Vienna and a co-author on the paper – explained in a University of Vienna press release:

“We were able to show that due to its metal oxidizing metabolic activity, when given an access to these Martian regolith simulants, M. sedula actively colonizes them, releases soluble metal ions into the leachate solution and alters their mineral surface leaving behind specific signatures of life, a ‘fingerprint’, so to say.”

Microspheroids containing mostly aluminium and chlorine overgrow the mineral surface of synthetic Mars regolith. These microspheroids can only be observed after cultivation of Metallosphaera sedula Credit: Tetyana Milojevic

The team then examined the samples of regolith to see if they had undergone any bioprocessing, which was possible thanks to the assistance of Veronika Somoza – a chemist from the University of Vienna’s Department of Physiological Chemistry and a co-author on the study. Using an electron microscope, combined with analytical spectroscopy technique, the team sought to determine if metals with the samples had been consumed.

In the end, the sets of microbiological and mineralogical data they obtained showed signs of free soluble metals, which indicated that the bacteria had effectively colonized the regolith samples and metabolized some of the metallic minerals within. As Milojevic indicated:

“The obtained results expand our knowledge of biogeochemical processes of possible life beyond Earth, and provide specific indications for detection of biosignatures on extraterrestrial material – a step further to prove potential extra-terrestrial life.”

In effect, this means that extreme bacteria could have existed on Mars billions of years ago. And thanks to the state of Mars today – with its thin atmosphere and lack of precipitation – the biosignatures they left behind (i.e. traces of free soluble metals) could be preserved within Martian regolith. These biosignatures could therefore be detected by upcoming sample-return missions, such as the Mars 2020 rover.

Biotransformed synthetic Martian Regolith after Metallosphaera sedula cultivation. Credit: Tetyana Milojevic

In addition to pointing the way towards possible indications of past life on Mars, this study is also significant as far as the hunt for life on other planets and star systems is concerned. In the future, when we are able to study extra-solar planets directly, scientists will likely be looking for signs of biominerals. Among other things, these “fingerprints” would be a powerful indicator of the existence of extra-terrestrial life (past or present).

Studies of extreme lifeforms and the role they play in the geological history of Mars and other planets is also helpful in advancing our understanding of how life emerged in the early Solar System. On Earth too, extreme bacteria played an important role in turning the primordial Earth into a habitable environment, and play an important role in geological processes today.

Last, but not least, studies of this nature could also pave the way for biomining, a technique where strains of bacteria extract metals from ores. Such a process could be used for the sake of space exploration and resource exploitation, where colonies of bacteria are sent out to mine asteroids, meteors and other celestial bodies.

Further Reading: University of Vienna, Frontiers in Microbiology

Europa’s Acidic Oceans May Prohibit Life

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The more we explore our solar system, the more we find things in common. Jupiter’s frigid moon – Europa – is about the size of our satellite and – like Earth – home to some very hostile environments. Underneath what is surmised to be an icy crust a few miles deep, Europa may possess an acidic ocean that could extend down as much as 100 miles (160 km) below the surface. We know from exploring our home planet that life happens under some very extreme conditions here… But what about Europa? What are the chances that life could exist there, too?

Check out liquid water on Earth and you’ll find some form of life. As a given, scientists hypothesize other worlds which contain water should also support life. According to recent studies, Europa’s ocean might even be saturated with oxygen – further supporting these theories. However, there’s a catch. Like Earth, surface chemicals are continually drawn downward. According to researcher Matthew Pasek, an astrobiologist at the University of South Florida, this could constitute a highly acidic ocean which “is probably not friendly to life — it ends up messing with things like membrane development, and it could be hard building the large-scale organic polymers.”

According to Charles Choi of Astrobiology Magazine, “The compounds in question are oxidants, which are capable of receiving electrons from other compounds. These are usually rare in the solar system because of the abundance of chemicals known as reductants such as hydrogen and carbon, which react quickly with oxidants to form oxides such as water and carbon dioxide. Europa happens to be rich in strong oxidants such as oxygen and hydrogen peroxide which are created by the irradiation of its icy crust by high-energy particles from Jupiter.”

Although it’s speculation, if Europa produces oxidants, they may also be drawn toward its core from ocean motion. However, it might be infused with sulfides and other compounds creating sulfuric and other acids before supporting life. According to the researchers, if this has happened for just half of Europa’s lifetime, the result would be corrosive, with a pH of about 2.6, “about the same as your average soft drink,” Pasek said. While this wouldn’t prohibit life from forming, it wouldn’t make it easy. Emerging life forms would have to be quick to consume oxidants and build an acid tolerance – a process which could take as much as 50 million years.

Are there similar acid-lovin’ lifeforms on Earth? You bet. They exist in acid mine drainage found in Spain’s Rio Tinto river and they feed on iron and sulfide for their metabolic energy. “The microbes there have figured out ways of fighting their acidic environment,” Pasek said. “If life did that on Europa, Ganymede, and maybe even Mars, that might have been quite advantageous.” It is also possible that sediments at the bottom of Europa’s ocean may neutralize the acids, even though Pasek speculates this isn’t likely. One thing we do know about an acidic ocean is that it dissolves calcium-based materials such as bones and shells.

It’s a lesson repeated on Earth…

Right now our oceans are absorbing excess carbon dioxide from the air which – when combined with seawater – forms carbonic acid. While it is mostly neutralized by fossil carbonate shells at the ocean’s bed, if it’s absorbed too quickly it can have some major ramifications on sea life such as coral reefs, plankton and mollusks. According to a recent study, this acidification is happening faster (thanks to human carbon emissions) than it has during four major extinction events on Earth in the last 300 million years.

“What we’re doing today really stands out,” said lead author Bärbel Hönisch, a paleoceanographer at Columbia University’s Lamont-Doherty Earth Observatory. “We know that life during past ocean acidification events was not wiped out—new species evolved to replace those that died off. But if industrial carbon emissions continue at the current pace, we may lose organisms we care about—coral reefs, oysters, salmon.”

According to this new research, our carbon dioxide levels have escalated by 30% in the last century. This means we’ve jumped to to 393 parts per million, and ocean pH has fallen by 0.1 unit, to 8.1–an acidification rate at least 10 times faster than 56 million years ago, says Hönisch. If this continues, the Intergovernmental Panel on Climate Change predicts the pH may drop as much as another 0.3 units… a drop that will constitute major biologic changes. While you might scoff at the extinction of a few forms of plankton or the annihilation of a small coral or shellfish, there is a ripple effect that cannot be denied.

“It’s not a problem that can be quickly reversed,” said Christopher Langdon, a biological oceanographer at the University of Miami who co-authored the study on Papua New Guinea reefs. “Once a species goes extinct it’s gone forever. We’re playing a very dangerous game.”

It may take decades before ocean acidification’s effect on marine life shows itself. Until then, the past is a good way to foresee the future, says Richard Feely, an oceanographer at the National Oceanic and Atmospheric Administration who was not involved in the study. “These studies give you a sense of the timing involved in past ocean acidification events—they did not happen quickly,” he said. “The decisions we make over the next few decades could have significant implications on a geologic timescale.”

For now, we’ll look to Europa and wonder at what may exist below its frozen waves. Is there an acid-loving form of life just waiting to bubble to the surface for us to find? Right now researchers are developing a drill which could assist in looking for extreme forms of life. The “penetrator” could eventually be part of a Europa exploration mission which could begin as early as 2020.

“Penetrators are the most feasible, cheapest and safest option for a landing on Europa today, and the knowledge to build those is there,” said Peter Weiss, a post-doc now at the National Center for Scientific Research (CNRS) in France. “Otherwise, we won’t have any confirmation on astrobiology on Europa — or maybe even in the solar system — during our lifetime.”

Original Story Source: Astrobiology Magazine. For Further Reading: Physorg.com.

Mars Likely Not Ever Warm and Wet Enough for Life – At Least on Surface

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Mars’ surface was probably not ever warm and wet long enough to support life, a new study published today in Nature concludes. But underground on the Red Planet might be a different story. By taking a look at several years of data from orbiting spacecraft and examining more than 350 sites on Mars, a team of researchers determined that Martian environments with abundant liquid water on the surface existed only in short episodes. But liquid and likely warm water more likely lasted for longer periods of time below the surface, and this would have been occurring at about the same time that life was developing on Earth.

“If surface habitats were short-term, that doesn’t mean we should be glum about prospects for life on Mars, but it says something about what type of environment we might want to look in,” said Bethany Ehlmann from Caltech and JPL, who is the lead author of the study. “The most stable Mars habitats over long durations appear to have been in the subsurface. On Earth, underground geothermal environments have active ecosystems.”

And so, the best place to look for signs of past life on Mars may be underground.

The researchers’ findings seem to indicate that Mars’ surface was almost always cold and dry, and any appearances of water – and the salts they left behind – occurred during geologically brief periods. This is certainly not the first time research has suggested brief periods of water flowing on Mars, or that underground water may have persisted, but the news study does help to provide a better picture of the history of water on Mars and even if it could possibly be there today.

Clays are crucial to understanding past water on Mars, as they form only when water is around long enough to change the chemical structure of rocks into clay, and different types of clay minerals result from different types of wet conditions.

Signs of deep water, deep life? Erosion has exposed clays (light blue) that subterranean waters favorable to life may have formed eons ago in the Nili Fossae region of Mars. Credit: NASA/JPL/JHUAPL/University of Arizona/Brown University

In 2005, clay minerals were discovered in many regions of Mars by the OMEGA spectrometer on the ESA’s Mars Express. This finding seemed to indicate the planet was once warm and wet. But there’s a problem with Mars’ atmosphere – it is not thick enough now for water to be retained on Mars’ surface, and there is not scientific consensus that it was ever thick enough in the past to have allowed water to remain on the surface.

But this new study supports an alternative hypothesis that warm water persisted under Mars surface and many erosional features seen by the orbiting spacecraft were carved during brief periods when liquid water was stable at the surface.

“The types of clay minerals that formed in the shallow subsurface are all over Mars,” said John Mustard, professor at Brown University in Providence, R.I. Mustard a co-author of the study. “The types that formed on the surface are found at very limited locations and are quite rare.”

During the past five years, researchers used OMEGA and NASA’s Compact Reconnaissance Imaging Spectrometer, or CRISM, instrument on the Mars Reconnaissance Orbiter to identify clay minerals at thousands of locations on Mars. Clay minerals that form with small amounts of water usually retain the same chemical elements as those found in the original volcanic rocks later altered by the water.

The study interprets this to be the case for most terrains on Mars with iron and magnesium clays. In contrast, surface environments with higher ratios of water to rock can alter rocks further. Soluble elements are carried off by water, and different aluminum-rich clays form.

Another clue is detection of a mineral called prehnite. It forms at temperatures above about 400 degrees Fahrenheit (about 200 degrees Celsius). These temperatures are typical of underground hydrothermal environments rather than surface waters.

Two upcoming missions will help decipher the water clues left behind on Mars. The Curiosity rover, or the Mars Science Laboratory will be heading towards Gale Crater, to investigate a large, layered hill that contain clay and sulfate minerals. Curiosity is scheduled to launch later this month.

These new findings also have implications for how Mars’ atmosphere may have evolved over time, and the Mars Atmosphere and Volatile Evolution Mission, or MAVEN, in development for a 2013 launch, may provide evidence for or against this new interpretation of the Red Planet’s environmental history. This new study predicts MAVEN findings will beconsistent with the atmosphere not having been thick enough to provide warm, wet surface conditions for a prolonged period.

Source: JPL

Even the Early Universe Had the Ingredients for Life

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For us carbon-based life forms, carbon is a fairly important part of the chemical makeup of the Universe. However, carbon and oxygen were not created in the Big Bang, but rather much later in stars. How much later? In a surprising find, scientists have detected carbon much earlier in the Universe’s history than previously thought.

Researchers from Ehime University and Kyoto University have reported the detection of carbon emission lines in the most distant radio galaxy known. The research team used the Faint Object Camera and Spectrograph (FOCAS) on the Subaru Telescope to observe the radio galaxy TN J0924-2201. When the research team investigated the detected carbon line, they determined that significant amounts of carbon existed less than a billion years after the Big Bang.

How does this finding contribute to our understanding of the chemical evolution of the universe and the possibilities for life?

To understand the chemical evolution of our universe, we can start with the Big Bang. According to the Big Bang theory, our universe sprang into existence about 13.7 billion years ago. For the most part, only Hydrogen and Helium ( and a sprinkle of Lithium) existed.

So how do we end up with everything past the first three elements on the periodic table?

Simply put, we can thank previous generations of stars. Two methods of nucleosythesis (element creation) in the universe are via nuclear fusion inside stellar cores, and the supernovae that marked the end of many stars in our universe.

Over time, through the birth and death of several generations of stars, our universe became less “metal-poor” (Note: many astronomers refer to anything past Hydrogen and Helium as metals”). As previous generations of stars died out, they “enriched” other areas of space, allowing future star-forming regions to have conditions necessary to form non-star objects such as planets, asteroids, and comets. It is believed that by understanding how the universe created heavier elements, researchers will have a better understanding of how the universe evolved, as well as the sources of our carbon-based chemistry.

So how do astronomers study the chemical evolution of our universe?

By measuring the metallicity (abundance of elements past Hydrogen on the periodic table) of astronomical objects at various redshifts, researchers can essentially peer back into the history of our universe. When studied, redshifted galaxies show wavelengths that have been stretched (and reddened, hence the term redshift) due to the expansion of our universe. Galaxies with a higher redshift value (known as “z”) are more distant in time and space and provide researchers information about the metallicity of the early universe. Many early galaxies are studied in the radio portion of the electromagnetic spectrum, as well as infra-red and visual.

The research team from Kyoto University set out to study the metallicity of a radio galaxy at higher redshift than previous studies. In their previous studies, their findings suggested that the main era of increased metallicity occurred at higher redshifts, thus indicating the universe was “enriched” much earlier than previous believed. Based on the previous findings, the team then decided to focus their studies on galaxy TN J0924-2201 – the most distant radio galaxy known with a redshift of z = 5.19.

The deep optical spectrum of TN J0924-2201 obtained with FOCAS on the Subaru Telescope. The red arrows point to the carbon emission line.

The research team used the FOCAS instrument on the Subaru Telescope to obtain an optical spectrum of galaxy TN J0924-2201. While studying TN J0924-2201, the team detected, for the first time, a carbon emission line (See above). Based on the detection of the carbon emission line, the team discovered that TN J0924-2201 had already experienced significant chemical evolution at z > 5, thus an abundance of metals was already present in the ancient universe as far back as 12.5 billion years ago.

If you’d like to read the team’s findings you can access the paper Chemical properties in the most distant radio galaxy – Matsuoka, et al at: http://arxiv.org/abs/1107.5116

Source: NAOJ Press Release

Citizen Science: Help Find Life on Mars

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Interested in helping NASA scientists pinpoint where to look for signs of life on Mars?

If so, you can join a new citizen science website called MAPPER, launched in conjunction with the Pavilion Lake Research Project’s 2011 field season.

How can the MAPPER and Pavilion Lake Research projects help scientists look for off-Earth life?

Since 2008, the Pavilion Lake Research Project (PLRP) has used DeepWorker submersible vehicles to investigate the underwater environment of two lakes in Canada (Pavilion and Kelly). With the MAPPER project, citizen scientists can work with NASA scientists and explore the lake bottoms from the view of a DeepWorker pilot.

The PLRP team’s main area of focus are freshwater carbonate formations known as microbialites. By studying microbialites that thrive in Pavilion and Kelly Lake, the scientists believe a better understanding of how the formations develop. Through a greater understanding of the carbonate formations, the team believes they will gain deeper insights into where signs of life may be found on Mars and beyond.

To investigate the formations in detail, video footage and photos of the lake bottom are recorded by DeepWorker sub pilots. The data requires analysis in order to determine what types of features can be found in different parts of the lake. Analyzing the data allows the team to answer questions such as; “how does microbialite texture and size vary with depth?” and “why do microbialites grow in certain parts of the lake but not in others?”.

The amount of data to analyze is staggering – if each image taken were to be printed, the stack would be taller than the depth of Pavilion Lake (over 60 meters). If each image were reviewed one-by-one, the PLRP’s team would never be able to complete their work. Distributing the work to the general public solves the problem, due in part by spreading the massive work out over many volunteers across the Internet.

Since the PLRP 2011 field season Morphology Analysis Project for Participatory Exploration and Research (MAPPER) MAPPER has been open to the general public. By opening MAPPER to the public, anyone can explore Pavilion and Kelly Lake as full-fledged members of PLRP’s Remote Science Team.

So how do volunteers use MAPPER to help the PLRP team?

Once volunteers create an account at: getmapper.com, the volunteers complete a brief tutorial, which provides the necessary training to tag photos in the PLRP dataset. MAPPER has ease-of-use in mind, providing users with a simple interface, which makes tagging features like sediment, microbialites, rocks, and algae easy. In case a user is unsure of how to tag a photo, examples and descriptions of each feature are available.

Screenshot of Mapper in action. Image Credit: NASA

In a manner similar to online games, each photo tagged earns the volunteer points which can be used to unlock new activities. Volunteers can also compete with other Remote Science Team members on the MAPPER leaderboard. Volunteers can also check to see how close each dataset is to being completely reviewed and see how much they have contributed to said dataset, as well as seeing what features have been tagged the most. Volunteers who tag a photo as ‘cool’ save said image to their Cool Photos album, allowing them to easily find the image at a later date.

PLRP Remote Science Team members from across North America, Europe and Asia have already been making discoveries in Pavilion and Kelly Lake. If you’d like to become a PLRP Remote Science Team member, visit: www.getmapper.com
You can also learn more by visiting the MAPPER Facebook page

All-Student Crew Lands at Mars Research Station

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Headline from the future? Actually, it’s happening now, although not quite on Mars, but about as close as humans can currently get. Six college students are the latest crew to embark on a two-week stint at the Mars Desert Research Station, a simulated Mars habitat set up by the Mars Society located in the San Rafael Swell of Utah. Looking across the very Mars-like red, rocky, panoramic vistas outside the habitat, participants might think they are on the Red Planet. And this latest crew, the 99th for MDRS, will be testing a microbial detection system and an EVA optimization method using an iPad.

The MDRS Campus in Utah, with the Habitat, observatory and greenhab.

The students — all graduate students or about to be – are from different colleges but came together in the summer of 2010 at the NASA Academy at the Ames Research Center in California, a 10-week immersive research internship.

“At the NASA Academy, we worked on a group project called LAMBDA – the Life and Microbial Detection Apparatus,” participant Max Fagin, from Dartmouth University, told Universe Today. “We wanted to do some follow-up work, in looking at microbial fuel cells, which run off the metabolic activity of bacteria — technology that could be applied to sewage reclamation plants in order to generate power.”

Fagin said the technology has been around a while, but they are trying to adapt it to detect microbes in soil samples, similar to what the Viking mission did in the 1970’s.

“We put a sample into the device and based on the power that is generated you can determine whether that power is coming from microbial activity or organic activity,” Fagin said.

They finished the summer internship with a good theoretical analysis and a non-working prototype, but wanted to field test their research, as well as continue work on other individual projects.

Crew patch for Crew LAMBDA.

Donna Viola, a senior undergraduate at the University of Maryland, Baltimore County, had been on two crew rotations on the MDRS previously and suggested to her fellow NASA Academy team that they apply as a group to the MDRS where they could test LAMBDA in actual conditions, with actual soil samples in the field where there may be potentially extremophile forms of life to find.

The team was accepted and began their crew rotation at MDRS on January 29. They will be there until February 12, all the while in complete Mars simulation. Crew members must wear a space suit when going outside the Habitat; they eat only space-travel type food (along with vegetables grown on-site in a greenhouse); power is provided by batteries or a power generation system; and there is also a water recycling system.

Viola is the Commander, Heidi Beemer is the team geologist and Executive Commander, Kevin Newman is the Engineer, Andie Gilkey is the team scientist and Health and Safety Officer, Chief Biologist, Sukrit Ranjan is the team astronomer and Fagin is the EVA Engineer.

See the crew biographies.

14 students total were part of the NASA Ames Academy, and even though only 6 are at the MDRS, the rest are serving as ground and mission support.

The last six weeks the team has been updating the LAMBDA device and making it field worthy, integrating it with the control system, and testing it.

While at MDRS, the crew has a few other projects, such as working on a proposed combination EVA planner and EVA monitor that runs on an iPad. “It monitors the astronauts’ health, vital signs, how much energy they are consuming, whether they should speed up or slow down – it’s basically an EVA optimizer,” Fagin said.

The Musk Observatory located at the Mars Desert Research Station

They will also fly a payload on a high altitude balloon that tests the feasibility of using balloon borne payloads on Mars. “There are no FAA regulations on Mars, so on Mars you could build a weather station on a balloon – such as on a 10 km tether and reel it in and out to get very nice vertical cuts of the atmospheric profiles of wind velocity and direction and dust profiles,” Fagin explained. “And also you could do astronomy by launching a small telescope. But we can’t do the tether part because they are here on Earth so we’ll be using a balloon and have to retrieve it.” They will also be flying a generic meteorological payloads and doing astronomical projects at the observatory on site, the Musk Observatory, which has a 14-inch telescope.

During their stay, the crew is required to send daily reports and dispatches from the commander, engineers, crew scientists, and journalists through the MDRS website which provides updates on the status of science experiments, updates on crew health and morale, and on the habitat and how it is faring. There is also a live webcam of different parts of the station.

MDRS is the second research station to be built by the Mars Society. The first was the Arctic station (FMARS) on Devon Island, built in 2000. Stations to be built in Europe (European Mars Analog Research Station / Euro MARS) and Australia (Australia Mars Analog Research Station / MARS Oz) are currently in the planning stages.

The goal of these analog research stations is to develop key knowledge, field tactics and equipment needed to prepare for the human exploration of Mars, testing habitat design features and tools, and to assess crew selection protocols. Utah is much warmer than Mars, the desert location is optimal because of its Mars-like terrain and appearance.

Find out more information on participating in the MDRS.

The first dispatches from the LAMBDA crew report how they are getting acclimated to the habitat and the equipment, as well as preparing for doing their actual science research.

Fagin said without the NASA Academy at Ames, this group of students wouldn’t be together at the MDRS today.

“This grew out of everything we did at the NASA Academy,” he said. “Without those experiences we would have no idea how to approach the situation, wouldn’t understand the science or engineering that needs to go into such a project, and certainly wouldn’t have the team-working abilities to do this if we hadn’t developed them while we were at the NASA Academy.”

Learn more about the NASA Ames Academy.

Universe Today hopes to provide an update on the LAMBDA crew’s activities.