Unit 1 Notes The Science of Microbiology “The microbe that felled one child in a distant continent yesterday can reach yours today and seed a global pandemic tomorrow.” Joshua Lederberg, Nobel Prize Winner, 1958 What Is Microbiology? Microbiology is defined as the study of microorganisms, or living things that are too small to be seen by the naked eye. Statistics Did you know that 25% of all deaths worldwide are due to infectious diseases (diseases caused by microbes)? The majority are respiratory infections, then AIDS, then diarrheal diseases. Why Do Microbes Matter? In spite of their tiny size, they have an enormous impact on our lives. Before the development of microbiology as a science, pathogens (microbes that cause human disease) often controlled human events. Microbes perform many functions in our lives. Here are some: o o o o o o o o Some cause diseases; some decompose dead organisms Some control pathogens in soil and maintain nature’s balance Some capture energy from the sun Some are used as insecticides and pesticides to control insects Some are used in the field of genetic engineering Some are used to make many of our food products, such as yogurt and beer Some are used to produce products we need such as insulin, medicines, etc. Some can be used to clean up oil spills and other toxic waste in the environment, called bioremediation The First Observations Bacterial ancestors were the first living cells to appear on Earth. Following are some of the first observations to be made in the discovery of microorganisms: In 1665, an Englishman named Robert Hooke was the first to notice that life's smallest living units were made up of "cells", which was the beginning of the cell theory (all living things are composed of cells). In the late 1674, Dutch scientist Antoni van Leeuwenhoek invented the first true microscope. He is the first to describe living microbes (he called them "animalcules"). The Debate Over Spontaneous Generation Until the late 1800's, people believed that life could arise spontaneously from nonliving matter (spontaneous generation). 1 In 1668, Francisco Redi did an experiment that demonstrated that maggots did not arise spontaneously from meat, and this was the beginning of the disproving of the spontaneous generation theory. In 1858, German scientist Rudolf Virchow challenged spontaneous generation with the concept of biogenesis, which says that living cells can only arise from other living cells. In 1859, however, Louis Pasteur settled the issue once and for all. In his experiments, he demonstrated conclusively that microbial life can be destroyed by heat. This was the beginning of aseptic technique, which is the use of techniques that prevent contamination by unwanted microorganisms, which is now standard practice in laboratories and medical facilities. He is credited with development of the "germ theory of disease", which states that germs, or microbes, are the causative agents of disease. The Golden Age of Microbiology The science of microbiology blossomed during a period of about 60 years, and this began in 1857 with Pasteur’s proposal of the germ theory of disease and continued into the 20th century, up until about World War I. This period was called the Golden Age of Microbiology. During these years, many branches of microbiology were established, and these findings led the way to modern microbiology, and many new disease and disease agents were discovered. In the 1930’s and 1940’s, the electron microscope was invented, and because a microbe can be multiplied about 100,000 times with these microscopes, viruses (the smallest microorganisms) were then discovered. At about the same time, antibiotics were discovered. After World War II, more money was available for research, and microbial genes were beginning to be studied. Here are some important discoveries during this time period: In 1796, Edward Jenner was the first to "vaccinate" people against smallpox by inoculating people with the pus from smallpox victims, thus immunizing them against the virus. This was before people even knew that microorganisms caused disease! The first real vaccines were made in the early 1880’s by Pasteur. In the 1860's, Joseph Lister, an English surgeon, was the first to apply the asceptic technique to medical procedures. He began treating surgical wounds with a phenol solution, which killed microorganisms. In 1876, Robert Koch, a German physician, proved that bacteria actually caused disease. He came up with a procedure to determine if a specific germ causes a specific disease, called Koch's postulates. In the late 1800's, Pasteur discovered fermentation (the process where yeast converts sugars to alcohol in the absence of air). He also discovered pasteurization (the process of heating food at low temperatures for a long time to kill bacteria). Microbiology Today After the Golden Age of Microbiology, advances in the field of microbiology continued to be made in 4 key areas: chemotherapy, immunology, virology, and genetic engineering. Chemotherapy – chemotherapy is the treatment of disease with chemicals called drugs. Now that the relationship between microbes and disease had been established, scientists now focused on finding substances that could cure disease. o In 1910, Paul Ehrlich, a German physician, created the first drug for the treatment of syphilis. He is called the father of modern chemotherapy because he was the first to 2 come up with the principle of selective toxicity (a successful drug must only kill the microbe, not the host). o In 1928, Alexander Fleming, a Scottish physician, discovered the first antibiotic called "penicillin" from mold. It wasn't until the 1940's that penicillin was finally massproduced. Immunology – this is the study of the immune system and how to develop vaccines to protect it. Virology – this is the study of viruses. The first virus was discovered in 1892 by Dmitri Iwanowski, and it was the tobacco mosaic virus. Genetic Engineering – this is the process where researchers take the genes from one organism, manipulate them in the laboratory, and insert them into another organism. The Microbial World 1. Archaea – discovered in the 1970’s, these are the “ancient” bacteria. They are also called “extremists” because they survive in extreme conditions. No archaea are human pathogens. They are so different from normal bacteria that they have their own group. They thrive in conditions that mimic those found more than 3.5 billion years ago. Lack a true nucleus. Cell walls contain different kinds of amino acids and sugars than do bacteria, as well as different lipid structures. They are living fossils. They have been found in acid, in ice, and in salt where normal bacteria cannot grow. Here are the basic types: a. Thermophiles – love hot temperatures. Can survive over 110 degrees Celsius. b. Psychrophiles – love cold temperatures. Have been found at -10 degrees Celsius. c. Halophiles – love salty habitats. Can survive in up to 9% salt water (ocean water is 0.9%). d. Acidophiles – love acidic conditions. 2. Bacteria – unicellular organisms that are classified according to their shapes; they are found everywhere. They are found in air, soil, water, on plants and animals, etc. Can be heterotrophic or autotrophic. Most utilize oxygen, but some operate anaerobically. Can be beneficial or harmful. The most numerous organisms on the Earth! 3. Viruses – made of a core of nucleic acid with a protein coat, viruses cannot survive on their own. They need host to metabolize nutrients, produce and excrete wastes, move around, and reproduce. Can infect all types of cells, including bacterial, fungal, protozoa, plants, animals, and human. They are the simplest and tiniest of microbes – up to 10,000 times smaller than bacteria. Includes two subgroups: a. Viroids – like viruses, but contain only RNA with no protein coat. b. Prions – like viruses, but do not contain nucleic acids. Just made of protein, but can transmit disease (Mad Cow Disease). 4. Protists – eukaryotic cells that are a very diverse group. The word “protist” comes from the Greek word meaning “very first”, and this is so because the protists are considered to be the first eukaryotes to appear on the earth. Classified by whether they are most like plants, animals or fungi. Include amoebas and algae. 5. Fungi – range in size from single-celled organisms to multi-celled organisms. Major decomposers of organic matter on Earth. Distinguished from other microbes by their physical appearance and by how they obtain their food. They secrete enzymes to break down organic matter. Cannot undergo photosynthesis. Many cause human disease. 6. Helminths – multicellular eukaryotic animals that possess digestive, circulatory, nervous, excretory, and reproductive systems. Also known as worms, they are parasitic: they must live inside a host in order to reproduce. 3 Characteristic Bacteria Protists Fungi Helminths Cell type Size Cell Wall Reproduction Energy process Prokaryotic Microscopic Present Binary fission Heterotrophic Eukaryotic Microscopic Absent Sexual and asexual Heterotrophic Eukaryotic Macroscopic Present Sexual and asexual Heterotrophic Eukaryotic Macroscopic Absent Sexual and asexual Heterotrophic Timeline of Important Milestones in Microbiology 1875 Ferdinand J. Cohn contributes to the founding of the science of bacteriology. He publishes an early classification of bacteria using the genus name Bacillus for the first time. 1876 Robert Koch publishes a paper on his work with anthrax, pointing explicitly to a bacterium as the cause of this disease. This validates the germ theory of disease. His work on anthrax was presented and his papers on the subject were published under the auspices of Ferdinand Cohn. 1884 tubercle proved by cultivation tuberculosis Medicine or Robert Koch puts forth a set of postulates, or standards of proof, involving the bacillus. Koch's postulates are published in The Etiology of Tuberculosis, in which he demonstrated three major facts: 1) the presence of the tubercule bacillus (as staining) in tubercular lesions of various organs of humans and animals, 2) the of the organisms in pure culture on blood serum, and 3) the production of at will by its inoculation into guinea pigs. Koch was awarded the Nobel Prize in Physiology in 1905. 1890 Sergei Winogradsky succeeds in isolating 1890-1891, Winogradsky performs the the process of nitrification in nature. nitrifying bacteria from soil. During the period definitive work on the organisms responsible for 1891 Paul Ehrlich proposes that antibodies are responsible for immunity. He shows that antibodies form against the plant toxins ricin and abrin. With Metchnikoff, Ehrlich awarded the Nobel Prize in Medicine or Physiology in 1908. is jointly 4 1892 Dmitri Ivanowski publishes the first evidence of the filterability of a pathogenic agent, the virus of tobacco mosaic disease, launching the field of virology. He passes the agent through candle filters that retain bacteria, but he isn't sure that the agent is a unique organism. 1912 Paul Ehrlich announces the first specific chemotherapeutic derivative and finally the 606th London, where Alexander discovery of an effective cure (Salvarsan) for syphilis, the agent for a bacterial disease. Ehrlich was seeking an arsenic compound worked. He brought news of the treatment to Fleming became one of the few physicians to administer it. 1915 Frederick Twort announces the first discovery of bacteriophages, or bacteria-infecting viruses. Twort’s discovery was something of an accident. He had spent several years growing viruses and noticed that the bacteria infecting his plates became transparent, indicating that they had been lysed or broken open and destroyed. Felix d’Herrelle independently describes bacterial viruses and coins the term “bacteriophage.” 1928 Frederick Griffith discovers transformation in bacteria and establishes the foundation of molecular genetics. He shows that injecting mice with a mixture of live, avirulent, rough Streptococcus pneumoniae Type I and heat-killed, virulent smooth S. pneumoniae Type II, leads to the death of the mice. Live, virulent, smooth S. pneumoniae Type II are isolated from the dead mice. 1929 Alexander Fleming publishes the first paper describing penicillin and its effect on grampositive microorganisms. This finding is unique since it is a rare example of bacterial lysis and not just microbial antagonism brought on by the mold Penicillium. Fleming kept his cultures 23 weeks before discarding them. When he looked at one set he noticed that the bacteria seemed to be dissolving and the mold was contaminating the culture. When penicillin is finally produced in major quantities in the 1940s, its power and availability effectively launch the “Antibiotics Era,” a major revolution in public health and medicine. With Florey and Chain, Fleming is awarded the Nobel Prize in Medicine or Physiology in 1945. 1935 Gerhard J. Domagk uses a chemically synthesized anti-metabolite, Prontosil, to kill Streptococcus in mice. One of the first patients to be treated with Protonsil is Domagk’s daughter who has a streptococcal infection that is unresponsive to other treatments. Near death, she is injected with large quantities of Protonsil and makes a dramatic recovery. Domagk is awarded the Nobel Prize in Medicine or Physiology in 1939. 5 1944 Oswald Avery, Colin MacLeod, and Maclyn McCarty show that DNA is the transforming material in cells. They demonstrate that the transformation of Streptococcus pneumoniae from an avirulent type to a virulent type is the result of the transfer of DNA from dead smooth organisms to live rough ones. They also show that the transforming principle is destroyed by pancreatic deoxyribonuclease —an enzyme that hydrolyzes DNA —but is not affected by pancreatic ribonuclease or enzymes that destroy proteins. 1944 Albert Schatz, E. Bugie and Selman Waksman used against tuberculosis. Streptomycin has the same negative microorganisms as penicillin does on gramthe Nobel Prize in Medicine or Physiology in 1952. discover streptomycin, soon to be specific antibiotic effect against grampositive ones. Waksman is awarded 1946 Joshua Lederberg and Edward L. Tatum publish the first paper on a type of bacterial mating called conjugation. The proof is based on the generation of daughter cells able to grow in media that cannot support growth of either of the parent cells. Their experiments showed that this type of gene exchange requires direct contact between bacteria. At the time Lederberg began studying with Tatum, scientists believed that bacteria reproduced asexually, but from the work of Beadle and Tatum, Lederberg knew that fungi reproduced sexually and he suspected that bacteria did as well. 1949 Microbiologist John Franklin Enders, virologist Thomas H. Weller and physician Frederick Chapman Robbins together develop a technique to grow poliovirus in test cultures of human tissues. This approach gave virologists a practical tool for the isolation and study of viruses. Enders, Weller and Robbins were awarded the Nobel Prize in Medicine or Physiology in 1954. tube 1952 Joshua Lederberg and Norton Zinder report on transduction, or transfer of genetic information to cells by viruses. They show that a phage of Salmonella typhimurium can carry DNA from one bacterium to another. 1952 Alfred Hershey and Martha Chase suggest that only DNA is needed for viral replication. Using radioactive isotopes 35S to track protein and 32P to track DNA, they show that progeny T2 bacteriophage isolated from lysed bacterial cells have the labeled nucleic acid. Further, most of the labeled protein doesn’t enter the cells but remains attached to the bacterial cell membrane. 1960 Francois Jacob, David Perrin, Carmen Sanchez and Jacques Monod propose the operon concept for control of bacteria gene action. Jacob and Monod later propose that a protein repressor blocks RNA synthesis of a specific set of genes, the lac operon, unless an inducer, 6 lactose, binds to the repressor. With Lwoff, Jacob and Monod are awarded the Nobel Prize in Medicine or Physiology in 1965. 1973 Stanley Cohen, Annie Chang, Robert Helling and Herbert Boyer show that extrachromosomal bits of DNA called plasmids act as vectors for maintaining cloned genes in bacteria. They show that if DNA is broken into fragments and combined with plasmid DNA, such recombinant DNA molecules will reproduce if inserted into bacterial cells. The discovery is a major breakthrough for genetic engineering, allowing for such advances as gene cloning and the modification of genes. 1977 Carl Woese uses ribosomal RNA analysis to recognize a third form of life, the Archaea, whose genetic makeup is totally different from the regular bacteria. He is the creator of the three branch evolutionary tree. 1979 Smallpox (variola) is declared officially eliminated, the last naturally occurring case having been seen in 1977 in Somalia. Small quantities remain held under tightly controlled conditions in the U.S. and former U.S.S.R. Smallpox is the only microbial disease to ever have been deliberately eradicated. 1982 Stanley Prusiner finds evidence that disease can be caused by a class of infectious proteins he call prions. These abnormal proteins cause scrapie, a fatal neurodegenerative disease of sheep. Prusiner is awarded the Nobel Prize in Medicine or Physiology in 1997. 1983 Luc Montagnier and Robert Gallo announce their discovery of the immunodeficiency virus (HIV) believed to cause AIDS. 1986 Kary Mullis uses a heat stable enzyme from Thermus aquaticus to establish polymerase chain reaction technology. PCR is used to amplify target DNA many-fold. Mullis is awarded the Nobel Prize in Chemistry in 1993. 7 1995 Craig Venter, Hamilton Smith, Claire Fraser and colleagues at TIGR elucidate the first complete genome sequence of a microorganism: Haemophilus influenza. 8