Prokaryotic Diversity

Prokaryotic Diversity

The advent of DNA sequencing provided immense insight into the relationships and origins of prokaryotes that were not possible using traditional methods of classification. A major insight identified two groups of prokaryotes that were found to be as different from each other as they were from eukaryotes (bacteria and archaea). This recognition of prokaryotic diversity forced a new understanding of the classification of all life and brought us closer to understanding the fundamental relationships of all living things, including ourselves.

Early Life on Earth

When and where did life begin? What were the conditions on Earth when life began? Prokaryotes were the first forms of life on Earth, and they existed for billions of years before plants and animals even appeared. Earth is about 4.54 billion years old. This estimate is based on evidence from the dating of meteorite material, since surface rocks on Earth are not as old as Earth itself. Most rocks available on Earth have undergone geological changes that make them younger than Earth itself. Some meteorites are made of the original material in the solar disk that formed the objects of the solar system, and they have not been altered by the processes that altered rocks on Earth. Thus, the age of meteorites is a good indicator of the age of the formation of Earth. The original estimate of 4.54 billion years was obtained by Clair Patterson in 1956. His meticulous work has since been corroborated by ages determined from other sources, all of which point to an Earth age of about 4.54 billion years.

Early Earth had a very different atmosphere than it does today. Evidence indicates that during the first 2 billion years of Earth’s existence, the atmosphere was anoxic, meaning that there was no oxygen. Therefore, only those organisms that can grow without oxygen—anaerobic organisms—were able to live. Organisms that convert solar energy into chemical energy are called phototrophs. Phototrophic organisms that required an organic source of carbon appeared within one billion years of the formation of Earth. Then, cyanobacteria, also known as blue-green algae, evolved from these simple phototrophs one billion years later. Cyanobacteria are able to use carbon dioxide as a source of carbon. Cyanobacteria (Figure 10.2) began the oxygenation of the atmosphere. The increase in oxygen concentration allowed the evolution of other life forms.

Figure 10.2 This hot spring in Yellowstone National Park flows toward the foreground. Cyanobacteria in the spring are green, and as water flows down the heat gradient, the intensity of the color increases because cell density increases. The water is cooler at the edges of the stream than in the center, causing the edges to appear greener. (credit: Graciela Brelles-Mariño)

Before the atmosphere became oxygenated, the planet was subjected to strong radiation; thus, the first organisms would have flourished where they were more protected, such as in ocean depths or beneath the surface of Earth. At this time, too, strong volcanic activity was common on Earth, so it is likely that these first organisms—the first prokaryotes—were adapted to very high temperatures. These are not the typical temperate environments in which most life flourishes today; thus, we can conclude that the first organisms that appeared on Earth likely were able to withstand harsh conditions.

Microbial mats may represent the earliest forms of life on Earth, and there is fossil evidence of their presence, starting about 3.5 billion years ago. A microbial mat is a large multi-layered sheet of prokaryotes (Figure 10.3a), including mostly bacteria, but also archaea. Microbial mats are a few centimeters thick, and they typically grow on moist surfaces. Their various types of prokaryotes carry out different metabolic pathways, and for this reason, they reflect various colors. Prokaryotes in a microbial mat are held together by a gummy-like substance that they secrete.

The first microbial mats likely obtained their energy from hydrothermal vents. A hydrothermal vent is a fissure in Earth’s surface that releases geothermally heated water. With the evolution of photosynthesis about 3 billion years ago, some prokaryotes in microbial mats came to use a more widely available energy source—sunlight—whereas others were still dependent on chemicals from hydrothermal vents for food.

Figure 10.3 (a) This microbial mat grows over a hydrothermal vent in the Pacific Ocean. Chimneys such as the one indicated by the arrow allow gases to escape. (b) This photo shows stromatolites that are nearly 1.5 billion years old, found in Glacier National Park, Montana. (credit a: modification of work by Dr. Bob Embley, NOAA PMEL; credit b: modification of work by P. Carrara, NPS)

Some prokaryotes are able to thrive and grow under conditions that would kill a plant or animal. Bacteria and archaea that grow under extreme conditions are called extremophiles, meaning “lovers of extremes.” Extremophiles have been found in extreme environments of all kinds, including the depths of the oceans, hot springs, the Arctic and the Antarctic, very dry places, deep inside Earth, harsh chemical environments, and high radiation environments. Extremophiles give us a better understanding of prokaryotic diversity and open up the possibility of the discovery of new therapeutic drugs or industrial applications. They have also opened up the possibility of finding life in other places in the solar system, which have harsher environments than those typically found on Earth. Many of these extremophiles cannot survive in moderate environments.

Biofilms

Biofilms are tightly-knit microbial communities anchored to surfaces by a sticky matrix mainly composed of polysaccharides, along with some proteins and nucleic acids. Previously thought to exist largely in isolation, we now understand that most prokaryotes actually prefer these communal settings for better interaction and protection. These biofilms are incredibly pervasive, showing up everywhere from industrial pipes to kitchen counters, and are often implicated in large-scale bacterial contamination events, including food outbreaks. What makes biofilms especially tricky is their resilience. The communal structure and protective matrix render them far more robust and resistant to sterilization methods compared to their free-living, or planktonic, counterparts.

(Difference between microbial mats and biofilms: Microbial mats are multi-layered, more complex communities found in specific environments, featuring a diverse range of microorganisms with specialized roles. In contrast, biofilms are usually simpler microbial communities that form on surfaces and often consist of a single or few types of microorganisms).

Characteristics of Prokaryotes

There are many differences between prokaryotic and eukaryotic cells. However, all cells have four common structures: a plasma membrane that functions as a barrier for the cell and separates the cell from its environment; cytoplasm, a jelly-like substance inside the cell; genetic material (DNA and RNA); and ribosomes, where protein synthesis takes place (from RNA instructions). Prokaryotes come in various shapes, but many fall into three categories: cocci (spherical), bacilli (rod-shaped), and spirilla (spiral-shaped) (Figure 10.4).

Figure 10.4 Many prokaryotes fall into three basic categories based on their shape: (a) cocci, or spherical; (b) bacilli, or rod-shaped; and (c) spirilla, or spiral-shaped. (credit a: modification of work by Janice Haney Carr, Dr. Richard Facklam, CDC; credit c: modification of work by Dr. David Cox, CDC; scale-bar data from Matt Russell)

The Prokaryotic Cell

Recall that prokaryotes (Figure 10.5) are unicellular organisms that lack organelles surrounded by membranes. Therefore, they do not have a nucleus but instead have a single chromosome—a piece of circular DNA (imagine a closed loop, like a rubber band or a hair tie) located in an area of the cell called the nucleoid. Most prokaryotes have a cell wall lying outside the plasma membrane. The composition of the cell wall differs significantly between the domains Bacteria and Archaea (and their cell walls also differ from the eukaryotic cell walls found in plants and fungi.) The cell wall functions as a protective layer and is responsible for the organism’s shape. Some other structures are present in some prokaryotic species, but not in others. For example, the capsule found in some species enables the organism to attach to surfaces and protects it from dehydration. Some species may also have flagella (singular, flagellum) used for locomotion, and pili (singular, pilus) used for attachment to surfaces and to other bacteria for conjugation. Plasmids, which consist of small, circular pieces of DNA outside of the main chromosome, are also present in many species of bacteria.

Figure 10.5 The features of a typical bacterium cell are shown.

Both Bacteria and Archaea are types of prokaryotic cells. They differ in the lipid composition of their cell membranes and in the characteristics of their cell walls. Both types of prokaryotes have the same basic structures, but these are built from different chemical components that are evidence of an ancient separation of their lineages. The archaeal plasma membrane is chemically different from the bacterial membrane.

The cell wall is a protective layer that surrounds some prokaryotic cells and gives them shape and rigidity. It is located outside the cell membrane and prevents osmotic lysis (bursting caused by increasing volume). The chemical compositions of the cell walls vary between Archaea and Bacteria, as well as between bacterial species.

Reproduction

Prokaryotes primarily reproduce asexually through binary fission, a process that creates two identical cells without the involvement of mitosis. In binary fission, the single, circular chromosome is replicated, and the two copies move apart as the cell enlarges. Eventually, the cell pinches inwards, splitting into two clone cells. While binary fission itself doesn’t facilitate genetic recombination, prokaryotes can introduce genetic variability through three mechanisms: transformation, transduction, and conjugation. Transformation involves the uptake of external DNA from the environment, potentially altering characteristics like pathogenicity. Transduction is mediated by bacteriophages (viruses that infect bacteria) that transfer DNA between bacteria. In conjugation, a pilus (hair-like structure on the cell surface) facilitates the direct transfer of DNA, usually in the form of plasmids, from one prokaryote to another.

These mechanisms for genetic variability, combined with the rapid life cycle of prokaryotes, enable swift evolutionary responses. Some prokaryotes can reproduce in a matter of minutes, allowing for speedy adaptation to environmental changes. This fast turnover and potential for genetic recombination make prokaryotes particularly adaptable, for instance, in developing resistance to antibiotics.

How Prokaryotes Obtain Energy and Carbon

Prokaryotes are metabolically diverse organisms. Prokaryotes fill many niches on Earth, including being involved in nutrient cycles such as the nitrogen and carbon cycles, decomposing dead organisms, and growing and multiplying inside living organisms, including humans. Different prokaryotes can use different sources of energy to assemble macromolecules from smaller molecules. Phototrophs obtain their energy from sunlight. Chemotrophs obtain their energy from chemical compounds.

Bacterial Diseases in Humans

Devastating pathogen-borne diseases and plagues, both viral and bacterial in nature, have affected and continue to affect humans. It is worth noting that all pathogenic prokaryotes are Bacteria; there are no known pathogenic Archaea in humans or any other organism. Pathogenic organisms evolved alongside humans. In the past, the true cause of these diseases was not understood, and some cultures thought that diseases were a spiritual punishment or were mistaken about material causes. Over time, people came to realize that staying apart from afflicted persons, improving sanitation, and properly disposing of the corpses and personal belongings of victims of illness reduced their own chances of getting sick.

Historical Perspective

There are records of infectious diseases as far back as 3,000 B.C. A number of significant pandemics caused by Bacteria have been documented over several hundred years. Some of the largest pandemics led to the decline of cities and cultures. Many were zoonoses that appeared with the domestication of animals, as in the case of tuberculosis. A zoonosis is a disease that infects animals but can be transmitted from animals to humans.

Infectious diseases remain among the leading causes of death worldwide. Their impact is less significant in many developed countries, but they are important determiners of mortality in developing countries. The development of antibiotics did much to lessen the mortality rates from bacterial infections, but access to antibiotics is not universal, and the overuse of antibiotics has led to the development of resistant strains of bacteria. Public sanitation efforts that dispose of sewage and provide clean drinking water have done as much or more than medical advances to prevent deaths caused by bacterial infections.

In 430 B.C., the plague of Athens killed one-quarter of the Athenian troops that were fighting in the Great Peloponnesian War. The disease killed a quarter of the population of Athens in over 4 years and weakened Athens’ dominance and power. The source of the plague may have been identified recently when researchers from the University of Athens were able to analyze DNA from teeth recovered from a mass grave. The scientists identified nucleotide sequences from a pathogenic bacterium that causes typhoid fever.¹

From 541 to 750 A.D., an outbreak called the plague of Justinian (likely a bubonic plague) eliminated, by some estimates, one-quarter to one-half of the human population. The population in Europe declined by 50 percent during this outbreak. Bubonic plague would decimate Europe more than once.

One of the most devastating pandemics was the Black Death (1346 to 1361), which is believed to have been another outbreak of bubonic plague caused by the bacterium Yersinia pestis. This bacterium is carried by fleas living on black rats. The Black Death reduced the world’s population from an estimated 450 million to about 350 to 375 million. Bubonic plague struck London hard again in the mid-1600s. There are still approximately 1,000 to 3,000 cases of plague globally each year. Although contracting bubonic plague before antibiotics meant almost certain death, the bacterium responds to several types of modern antibiotics, and mortality rates from plague are now very low.

Over the centuries, Europeans developed resistance to many infectious diseases. However, European conquerors brought disease-causing bacteria and viruses with them when they reached the Western hemisphere, triggering epidemics that completely devastated populations of Native Americans (who had no natural resistance to many European diseases).

Fast-forward to the 20th and 21st centuries, and we see the devastating impact of viral pandemics like the Spanish flu of 1918 and COVID-19 in 2020. The Spanish flu, caused by the H1N1 influenza virus, infected an estimated one-third of the world’s population and resulted in the deaths of at least 50 million people worldwide. Similarly, the COVID-19 pandemic, caused by the novel coronavirus SARS-CoV-2, has had a sweeping global impact, infecting millions and causing significant loss of life and economic upheaval. Both pandemics highlight the unpreparedness of healthcare systems and the profound societal changes that can result from such outbreaks. While advances in medicine, such as vaccines and antiviral drugs, have improved our ability to combat these diseases, the challenges posed by rapidly spreading pandemics continue to be a focal point for public health initiatives.

The Antibiotic Crisis

The word antibiotic comes from the Greek anti, meaning “against,” and bios, meaning “life.” An antibiotic is an organism-produced chemical that is hostile to the growth of other organisms. Today’s news and media often address concerns about an antibiotic crisis. Are antibiotics that were used to treat bacterial infections easily treatable in the past becoming obsolete? Are there new “superbugs”—bacteria that have evolved to become more resistant to our arsenal of antibiotics? Is this the beginning of the end of antibiotics? All of these questions challenge the healthcare community.

One of the main reasons for resistant bacteria is the overuse and incorrect use of antibiotics, such as not completing a full course of prescribed antibiotics. The incorrect use of an antibiotic results in the natural selection of resistant forms of bacteria. The antibiotic kills most of the infecting bacteria, and therefore only the resistant forms remain. These resistant forms reproduce, resulting in an increase in the proportion of resistant forms over non-resistant ones.

Another problem is the excessive use of antibiotics in livestock. The routine use of antibiotics in animal feed promotes bacterial resistance as well. In the United States, 70 percent of the antibiotics produced are fed to animals. The antibiotics are not used to prevent disease, but to enhance production of their products.

Staphylococcus aureus, often called “staph,” is a common bacterium that can live in and on the human body, which usually is easily treatable with antibiotics. A very dangerous strain, however, has made the news over the past few years (Figure 10.7). This strain, methicillin-resistant Staphylococcus aureus (MRSA), is resistant to many commonly used antibiotics, including methicillin, amoxicillin, penicillin, and oxacillin. While MRSA infections have been common among people in healthcare facilities, it is appearing more commonly in healthy people who live or work in dense groups (like military personnel and prisoners). The Journal of the American Medical Association reported that, among MRSA-afflicted persons in healthcare facilities, the average age is 68 years, while people with “community-associated MRSA” (CA-MRSA) have an average age of 23 years.²

Figure 10.7 This scanning electron micrograph shows methicillin-resistant Staphylococcus aureus bacteria, commonly known as MRSA. (credit: modification of work by Janice Haney Carr, CDC; scale-bar data from Matt Russell)

In summary, society is facing an antibiotic crisis. Some scientists believe that after years of being protected from bacterial infections by antibiotics, we may be returning to a time in which a simple bacterial infection could again devastate the human population. Researchers are working on developing new antibiotics, but few are in the drug development pipeline, and it takes many years to generate an effective and approved drug.

Foodborne Diseases

Prokaryotes are everywhere: They readily colonize the surface of any type of material, and food is not an exception. Outbreaks of bacterial infection related to food consumption are common. A foodborne disease (colloquially called “food poisoning”) is an illness resulting from the consumption of food contaminated with pathogenic bacteria, viruses, or other parasites. Although the United States has one of the safest food supplies in the world, the Center for Disease Control and Prevention (CDC) has reported that “76 million people get sick, more than 300,000 are hospitalized, and 5,000 Americans die each year from foodborne illness.”³

The characteristics of foodborne illnesses have changed over time. In the past, it was relatively common to hear about sporadic cases of botulism, the potentially fatal disease produced by a toxin from the anaerobic bacterium Clostridium botulinum. A can, jar, or package created a suitable anaerobic environment where Clostridium could grow. Proper sterilization and canning procedures have reduced the incidence of this disease.

Most cases of foodborne illnesses are now linked to produce contaminated by animal waste. For example, there have been serious, produce-related outbreaks associated with raw spinach in the United States and with vegetable sprouts in Germany (Figure 10.8). The raw spinach outbreak in 2006 was produced by the bacterium E. coli strain O157:H7. Most E. coli strains are not particularly dangerous to humans, (indeed, they live in our large intestine), but O157:H7 is potentially fatal.

Figure 10.8 (a) Locally grown vegetable sprouts were the cause of a European E. coli outbreak that killed 31 people and sickened about 3,000 in 2010. (b) Escherichia coli are shown here in a scanning electron micrograph. The strain of E. coli that caused a deadly outbreak in Germany is a new one not involved in any previous E. coli outbreaks. It has acquired several antibiotic resistance genes and specific genetic sequences involved in aggregation ability and virulence. It has recently been sequenced. (credit b: Rocky Mountain Laboratories, NIAID, NIH; scale-bar data from Matt Russell)

All types of food can potentially be contaminated with harmful bacteria of different species. Recent outbreaks of Salmonella reported by the CDC occurred in foods as diverse as peanut butter, alfalfa sprouts, and eggs.

Beneficial Prokaryotes

Not all prokaryotes are pathogenic. On the contrary, pathogens represent only a very small percentage of the diversity of the microbial world. In fact, our life and all life on this planet would not be possible without prokaryotes.

Prokaryotes, and Food and Beverages

According to the United Nations Convention on Biological Diversity, biotechnology is “any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use.”⁴ The concept of “specific use” involves some sort of commercial application. Genetic engineering, artificial selection, antibiotic production, and cell culture are current topics of study in biotechnology. However, humans have used prokaryotes to create products before the term biotechnology was even coined. And some of the goods and services are as simple as cheese, yogurt, sour cream, vinegar, cured sausage, sauerkraut, and fermented seafood that contains both bacteria and archaea (Figure 10.9).

Figure 10.9 Some of the products derived from the use of prokaryotes in early biotechnology include (a) cheese, (b) salami, (c) yogurt, and (d) fish sauce. (credit b: modification of work by Alisdair McDiarmid; credit c: modification of work by Kris Miller; credit d: modification of work by Jane Whitney)

Cheese production began around 4,000 years ago when humans started to breed animals and process their milk. Evidence suggests that cultured milk products, like yogurt, have existed for at least 4,000 years.

Using Prokaryotes to Clean up Our Planet: Bioremediation

Microbial bioremediation leverages prokaryotes to eliminate various pollutants, from agricultural chemicals and toxic metals like selenium and arsenic to the hazardous mercury often found in pesticides and industrial byproducts. Specialized bacteria, such as Pseudomonas aeruginosa, can transform toxic forms of these substances into nontoxic variants. The technique is notably employed in cleaning up oil spills (Figure 10.10), where inorganic nutrients are added to encourage the growth of indigenous, hydrocarbon-degrading bacteria like Alcanivorax borkumensis. These bacteria can break down up to 80% of nonvolatile oil components within a year under optimal conditions. However, some oil fractions are more resistant to degradation and can linger in the environment. The first U.S. patent application for bioremediation even centered on a genetically engineered bacterium designed to consume petroleum products.

Figure 10.10 (a) Cleaning up oil after the Valdez spill in Alaska, the workers hosed oil from beaches and then used a floating boom to corral the oil, which was finally skimmed from the water surface. Some species of bacteria are able to solubilize and degrade the oil. (b) One of the most catastrophic consequences of oil spills is the damage to fauna. (credit a: modification of work by NOAA; credit b: modification of work by GOLUBENKOV, NGO: Saving Taman)