CHAPTER  1      THE MICROBIAL WORLD AND YOU

 

Microbes (microorganisms) are living things that are usually too small to be seen without magnification.

Microbes include:

·         Bacteria

·         Protozoa

·         Fungi

·         Microscopic algae

·         Viruses (these are different from the rest—question whether they can e said to be alive)

 

Out first thought is probably that these cause disease. We call them  germs.” A more scientific term is pathogenic (disease-causing) microbes. These do get a lot of attention, but many more microbes are harmless or even beneficial.

 

     Food chains in various bodies of water begin with microbes

     Nitrogen-fixing bacteria in soil

     Algae--carry out photosynthesis and produce O2

     Digestive tract—aid in digestion and produce vitamins, which we absorb (B vitamins & K)

     Bacteria are used in industrial processes to produce chemicals such as:

          Acetone

          Organic acids

          Enzymes

          Alcohols

          Drugs such as antibiotics

     Bacteria and other microbes are used in production of foods such as:

          Sauerkraut                                  Buttermilk

          Pickles                                         Cheese

          Vinegar                                       Yogurt

          Alcoholic beverages                   Bread

          Soy Sauce

Bacteria are used in modern genetic engineering to produce such products as:

          Insulin

          hGH

          Interferon

 

It is not easy for us to even imagine a world where microbes were completely unknown. Things that everyone today takes for granted were complete mysteries.

     Epidemics occurred and no one could imagine the cause

     Food spoilage

     No vaccines

     No antibiotics--in 1937, 80% of pneumonia cases were fatal

 

 

NAMING AND CLASSIFYING MICROORGANISMS

 

Our system of nomenclature (naming) of living organisms was established in 1735 by Carolus Linnaeus, a Swedish botanist.  Living organisms except bacteria are classified into the following groups:

  • Kingdom
  • Phylum
  • Class
  • Order
  • Family
  • Genus
  • Species (specific epithet)

The last 2 groups, genus and species, are known as the scientific name. Both parts must be underlined or in italics.  Genus name is a Latin noun and is capitalized, species name is a Latin adjective and is not capitalized. Examples: Staphylococcus aureus, Escherichia coli or Staphylococcus aureus and Escherichia coli.

 

In classifying bacteria, some of the groups may not be in common use, but the genus & species always will be. For minor differences among members of the same bacterial species, the designation of strain is used.

 

Linneaus’ original classifications were based on observation and “common sense.” Modern classifications are mostly based on study of DNA and RNA. These are more scientifically accurate, but the great majority of the original classifications stand.

 

GROUPS OF MICROBES

 

1. Bacteria:  tiny, one-celled microbes whose cellular characteristics differ from the cells of all other living things.  Bacterial cells are prokaryotic cells, while the cells of all other organisms are eukaryotic.  Prokaryotic cells are simpler cells.  A major difference is that they have no membrane separating the genetic material from the rest of the cell contents.  This group is also sometimes called eubacteria--our common everyday bacteria. Their cell walls are made of peptidoglycan, a huge molecule made of carbohydrate and protein. Bacterial cells come in 3 basic shapes: bacillus (rod), coccus (round), and spiral (corkscrew or curved).

 

2. Archaea: these are strange bacteria which share some characteristics with the eubacteria, but also have some major differences. Their appearance is very much the same, and their cells are also prokaryotic. However, this group does not contain peptidoglycan in their cell walls, and they thrive in extreme environments. Their metabolic pathways often differ greatly from those of eubacteria. As far as we know, this group does not cause disease in humans.

 

3. Fungi:   these organisms have eukaryotic cells, with a nuclear membrane separating the genetic material from the other cell contents.  Unicellular fungi are known as yeasts and are all microscopic.  Other fungi are multicellular.  Mushrooms are an example of large multicellular fungi.  Smaller multicellular fungi are known as molds.  Molds are made of long filaments called hyphae, which branch and intertwine and form a visible mass called a mycelium.  Fungi are plant-like but do not have chlorophyll and cannot carry on photosynthesis.

 

4. Protozoa:  unicellular, eukaryotic more animal-like organisms.  Protozoa are placed into 4 groups based on means of locomotion (movement).  They may be free-living or parasitic.

 

5. Algae:   photosynthetic eukaryocytes.  Some are multicellular and macroscopic.  In microbiology, the unicellular algae are studied.  Some of these are motile by flagella. Algae produce large amounts of oxygen as they carry on photosynthesis.

 

6. Viruses:  tiny entities which are acellular---no cellular structure.  A virus consists of a core of nucleic acid covered by a protein coat. Viruses cannot reproduce or carry out metabolic reactions on their own, but they invade living cells and reproduce using the mechanisms of the host cell.

 

7. Parasitic worms  (helminths):  these organisms are microscopic during at least some stages of their life cycles, but many also have macroscopic stages.  The two main groups are the flatworms and the roundworms

 

Linneaus placed all organisms into 1 or 2 kingdoms---plant or animal. This was the accepted standard for about 100 years. In the middle 1800’s, a German named Haeckel proposed 3 kingdoms---animal, plant and one called Protista to give a place for microorganisms. This stood for around another 100 years, until advancing knowledge made a new system the better choice. This was Whittaker’s 5 kingdom scheme. It  included:

     *1. Kingdom Monera---all the bacteria (prokaryocytes)

     *2. Kingdom Protista---unicellular eukaryocytes

     *3. Kingdom Fungi---the fungi---yeasts and molds

      4. Kingdom Plantae---plants and higher algae

      5. Kingdom Animalia---animals

 

Another scheme has been proposed, based on the idea that all bacteria are not similar enough to be in the same kingdom. The main person behind this plan is Dr. Carl Woese, who claims that there should be 3 main groups called domains, which are taxa higher than kingdom:

     1. Bacteria---regular everyday bacteria with peptidoglycan cell walls

     2. Archaea---unusual bacteria lacking peptidoglycan cell walls

     3. Eukarya---all others

 

 

 

HISTORY OF MICROBIOLOGY

 

The science of microbiology began approximately 300 years ago, although most developments have occurred in about the last 200 years. We will look at some important steps in the history of microbiology.

 

I.  DEVELOPMENT OF THE MICROSCOPE

Before we could study them, we had to be able to see them!

 

Our first record of magnifying lenses dates back to the 13th century. The first microscopes were developed in the early 1600’s, by two scientists, Jansen and Galileo.

 

In 1665,  Robert Hooke observed empty dead cells in a piece of cork. He named them cells because they reminded him of the little rooms in a monastery, which were also called cells. He never seemed to understand the importance of his discovery, but this was a beginning.

 

In the years 1673-1723, a Dutch merchant,  Antoni van Leeuwenhoek, first recorded observations of microbes. He built his own microscopes as a hobby and constantly worked to improve them, but his best instrument probably had a magnification of 200 or 300 X. He had no scientific education, but he constantly reported his findings to the British Royal Society,  so his remarkable work was recorded this way. He called the tiny creatures he observed “animalcules,” and today we can recognize bacteria and protozoa from his drawings. He is known as the Father of Microbiology.

 

 

 

II.  DEBATE OVER SPONTANEOUS GENERATION

 

Although it seems silly today, until the mid 1800’s many scientists believed some forms of life arose spontaneously. In fact, this was a hotly debated issue for many years.  The opposing theory was known as biogenesis—life from life, or all cells come from previously existing cells.

 

Spontaneous generation:

·  Toads and snakes-moist soil

·  Mice-rags and grain

·  Flies-from manure

·  Maggots-from meat

 

An Italian, Fransisco Redi, in 1668 proved maggots appeared only if flies could get to meat---beginning of the end of the theory for macroscopic creatures but now believers turned to microbes.

 

1745-Needham-boiled meat to kill microbes and left uncovered---soon microbes appeared---claimed this proved spontaneous  generation

 

1765-Spallanzani (Italian) claimed microbes come from air. He boiled meat and kept sealed-no microbes. Needham replied keeping air out removed some “vital force” microbes needed. Lavoisier soon proved that  O2 is  required for many life forms and this seemed to back up Needham’s argument.

 

The arguments dragged on:

 

1858-Rudolf Virchow (German) officially proposed the theory of biogenesis-cells arise only from pre-existing living cells and this included microbes. Many scientists agreed with him, but there were still those opposed.

 

In 1861 a French scientist, Louis Pasteur, carried out the final proof of biogenesis. He devised a special goose-neck flask which he used to overcome the last arguments.

 

 

 

 

 

 

 

 

 

 

 

 

 

III.  GOLDEN AGE OF MICROBIOLOGY  1857 – 1914

 

·         Microbiology was established as a science

·         Many discoveries by Pasteur, Koch, and others

·         Causative agents of many diseases were identified

·         Role of immunity began to be understood

·         Improved microscopes

·         Improved lab culturing techniques

·         Vaccines were developed

·         Gram stain and other staining techniques were developed

 

 

IV.  FERMENTATION

 

First, more about Louis Pasteur. He was a French chemist who lived from 1822 - 1895. Although his education was in chemistry and he set out to be a chemistry teacher, he made great contributions in microbiology, medicine, the wine industry, agriculture, etc. He is recognized as the man most responsible for making the connection between microbes and disease, which may be his most important contribution.

 

In the 1850’s, Pasteur was asked to determine why wine sometimes turned out badly and why good wine sometimes spoiled.  He proved fermentation was due to the action of microbes . Good results were due to “good” microbes, and if other  bad” microbes were present, they caused spoilage.  His solution was to carefully heat the grape juice just enough to kill all microbes without changing the character of the juice, and then add  good” microbes. This technique came to be called pasteurization.

 

 

V.  GERM THEORY OF DISEASE

 

As far back as recorded history, great epidemics of diseases such as bubonic plague, smallpox, and cholera have occurred, and for most of that time man had no idea of the cause.  In the 1300’s an outbreak of bubonic plague killed about 25 million people in Europe, including about half the population of France. In more recent time, the influenza pandemic of 1918 - 1919 killed about 20 million people worldwide.

 

Before infectious diseases were understood, illness was blamed on such things as divine punishment, “bad air,” “evil humors,” “bad blood,” demons, etc.  Treatments included bleeding, physics, shutting out air, burning feathers in the room, etc.

 

Over the years, some early scientists had proposed that tiny living organisms were the cause of disease. This included an Italian, Fracastoro, in 1547, and an Austrian, von Plenciz, in 1762. Even though these men were right, their ideas were not widely accepted at the time.

 

In 1865, Pasteur proved that the cause of a disease of silkworms (pebrine) was a protozoan organism. This was the beginning of the proof that all infectious disease was caused by microorganisms. Pasteur went on to find the causative agents for a number of animal and human diseases.

 

A German physician, Robert Koch, become very interested in Pasteur’s work and continued to identify causative microbes. In 1876, Koch proved that anthrax, a disease of cattle, was caused by a bacterium, Bacillus anthracis.  Koch realized that to work with microbes, a way to grow them in lab for study was needed. Koch and his associates did a lot of the work on culture techniques. By trial and error they found ‘recipes’ for solutions many microbes would grow well in. At first, all of these were liquids, but Koch wanted a solid medium. He first used slices of potato, but then discovered that adding a seaweed product called agar to his liquid media would make them solidify.

 

An associate of Koch, Richard Petri, invented the Petri dish.

 

Koch determined the criteria for proof of causative agents. This set of rules is known as Koch’s Postulates. Remember, a pathogen is a disease-causing microbe.

·         Pathogen must be present in all cases of disease

·         Pathogen must be isolated and grown in lab in pure culture

·         Pathogen from pure cultures must cause disease when inoculated into healthy, susceptible lab animal

·         Same pathogen must be isolated from the diseased lab animal

 

Using his postulates, Koch identified causative agents for tuberculosis, diphtheria, human cholera, typhoid, tetanus, and other diseases. The postulates are still applied today, although there are some limitations.

 

Even before day of proof of causative agents, some doctors suspected that “germs” could be transferred from one patient to another.

·         1840’s---Oliver Wendell Holmes in the US and Ignaz Semmelweis in Hungary both realized that puerperal fever (childbed fever) which we now know is a streptococcal infection, was being transferred from patient to patient by doctors, and that simple hand washing would greatly decrease the incidence

·         1860’s---Lister, an English surgeon, pioneered the use of phenol (carbolic acid) for surgical instruments, cleaning the operating room, and dressing for incisions.

 

 

VI.  VACCINATION

 

·         1796---Edward Jenner, an English physician, performed the first safe vaccinations in modern medicine. He discovered that if people were deliberately exposed to a mild disease, cowpox, they became immune to smallpox. He did not call it vaccination-Pasteur later gave this name, in honor of Jenner, using the Latin word for cow. Jenner’s process was a lucky accident, since cross-immunization—becoming immune to one disease also provides immunity to another—is rare.

·         1880-Pasteur discovered a vaccine for fowl cholera. This was an accident, but he figured out what was happening and set out to develop vaccines for human and animal diseases. His first successful vaccine was for anthrax. He also discovered a preventative treatment for rabies, which was used until recent years.

 

Since instances of cross immunization are rare, vaccines are mostly produced by weakening but not killing virulent microbes, using only selected parts of virulent microbes, or even genetic engineering.

 

 

VII.  BIRTH OF CHEMOTHERAPY

 

Chemotherapy---chemical treatment of disease. Types of chemicals used against microbes:

     Antibiotics---produced naturally by bacteria and fungi

     Synthetic drugs---produced totally in lab

     Semi-synthetic---we take the natural antibiotic and modify it

For all of these, the chemical must be more poisonous to microbe than host.

 

First chemotherapy—used substances from nature such as plant products (folk medicine)

     Cinchona bark—quinine for malaria

     Willow bark—aspirin

     Foxglove--digitalis

 

Paul Ehrlich-deliberately looked for a treatment for syphilis. In 1910 he discovered an arsenic compound, Salvarsan, and this is regarded as the beginning of modern chemotherapy.

 

1930’s--sulfa drugs

 

Antibiotics---Fleming discovered penicillin by accident in 1928. He did not realize importance, but about 10 years later 2 other scientists did. It began to be available in 1940’s.

 

 

MODERN DEVELOPMENTS IN MICROBIOLOGY

 

As we learned more, various branches of micro developed:

     Bacteriology---study of bacteria

     Mycology---study of fungi

     Parasitology--- study of protozoa and parasitic worms

 

We have not yet discovered all pathogens. Later in this chapter we will have a whole list of newly discovered pathogens and diseases.

 

     Immunology- began with Jenner 1796. Another big development was the discovery of phagocytosis by white blood cells, for which Elie Metchnikoff received a Nobel Prize. Study in this area has advanced rapidly in recent years, mainly due to AIDS virus.

 

     Virology—as study of bacteria continued, it began to be recognized that some pathogens could pass through a filter small enough to trap all known bacteria. The name given to this unknown agent was virus. Scientists were only able to actually see them after the electron microscope was developed in the 1940’s.

 

 

 

RECOMBINANT DNA—GENETIC ENGINEERING

 

Little progress in molecular genetics and molecular biology took place until we began studying these things in one-celled organisms in the 1940’s. Then rapid progress was made.

·         1941---1 gene 1 enzyme

·         1944---DNA was identified as hereditary material

·         1953--- Watson and Crick discovered structure of DNA

·         1960’s--- more about the  way DNA controls protein synthesis, discovery of mRNA, regulation of gene function, “cracked” the genetic code

·         1970’s---beginning of genetic engineering

 

 

Recombinant DNA- genes can be isolated and inserted into other cells, even from one species to another, and proteins still be made. This technique is currently used to produce hormones, vaccines, etc. The hope is that eventually we may be able to insert the needed gene into the cells of a person with a genetic defect---gene therapy.

 

 

MORE BENEFICIAL ACTIVITIES OF MICROBES:

·         Recycling vital elements-decomposition of wastes and dead plants and animals by microbes as well as O2   production by photosynthetic microbes

·         Sewage treatment-bacteria are used in the breakdown of sewage to recycle the water

·         Cleanup of the environment-bacteria can aid in cleanup of pollutants and toxic wastes by producing enzymes that break down these substances.  Some of these bacteria are even genetically engineered to clean up specific chemicals.  This is called bioremediation.

·         Enzymes produced by bacteria (Bacillus) are used in detergents are used to remove spots

·         Insect control-due to concern about toxic insecticides in the environment, there is great interest in developing microbes that affect only certain insect pests

 

BIOTECHNOLOGY AND GENETIC ENGINEERING

·         Biotechnology---commercial use of microorganisms to produce food, chemicals, etc.

·         The most recent form involves insertion of specific genes into cells to allow productions of new proteins- for example, Escherichia  coli  bacteria are now making human proteins such as insulin.

·         Diseases caused by genetic defects may one day be cured by insertion of the normal gene into the cells of the patient (gene therapy)

·         Many crops have been improved by genetic engineering—better yield, insect resistance, l;onger shelf life, etc.

 

MICROBES AND DISEASE

 

Although most microbes are harmless and some are even beneficial, a small number do cause disease.  These are pathogens (disease-causing) microbes. Some of these pathogens can be traced as far back as human history.  With the development of antibiotics, vaccines and other modern treatments, there was a time that we believed infectious disease could be eradicated.  Development of resistant bacteria and appearance of new diseases has ended that belief. Some emerging infectious diseases newly recognized or found in locations where previously unknown in recent years include:

·         Avian influenza A (H5N1)—also called bird flu

·         SARS

·         West Nile encephalitis

·         Bovine spongiform encephalopathy (mad cow disease)

·         Escherichia coli O157:H7

·         Invasive Group A strep (flesh-eating bacteria)

·         Ebola fever

·         Marburg virus

·         Hantavirus pulmonary syndrome

·         Cryptosporidium

·         AIDS