Microbiology PDF

Microbiology Microbiology is the scientific study of microorganisms, those being of unicellular, multicellular, or acellular. Microbiology encompasses numerous sub-disciplines including virology, bacteriology, protistology, mycology, immunology, and parasitology. Study of microorganisms, or microb...

Microbiology Microbiology is the scientific study of microorganisms, those being of unicellular, multicellular, or acellular. Microbiology encompasses numerous sub-disciplines including virology, bacteriology, protistology, mycology, immunology, and parasitology. Study of microorganisms, or microbes, a diverse group of generally minute simple life-forms that include bacteria, archaea, algae, fungi, protozoa, and viruses. The field is concerned with the structure, function, and classification of such organisms and with ways of both exploiting and controlling their activities. The 17th-century discovery of living forms existing invisible to the naked eye was a significant milestone in the history of science, for from the 13th century onward it had been postulated that "invisible" entities were responsible for decay and disease. The word microbe was coined in the last quarter of the 19th century to describe these organisms, all of which were thought to be related. As microbiology eventually developed into a specialized science, it was found that microbes are a very large group of extremely diverse organisms. Daily life is interwoven inextricably with microorganisms. In addition to populating both the inner and outer surfaces of the human body, microbes abound in the soil, in the seas, and in the air. Abundant, although usually unnoticed, microorganisms provide ample evidence of their presence sometimes unfavourably, as when they cause decay of materials or spread diseases, and sometimes favourably, as when they ferment sugar to wine and beer, cause bread to rise, flavour cheeses, and produce valued products such as antibiotics and insulin. Microorganisms are of incalculable value to Earth's ecology, disintegrating animal and plant remains and converting them to simpler substances that can be recycled in other organisms. Importance of Microbiology in Everyday Life 1. Importance of Microbiology in Food Industry: Microorganisms involved in food microbiology include bacteria molds and yeasts. Bacteria mainly cause food intoxication and food spoilage thereby causing various human gut health diseases. Several bacterial strains are used to produce a wide range of food and dairy products. These bacterial strains include Streptococcus thermophiles, Bifidobacterium sp, and Lactobacillus bulgaricus. Lactic acid bacteria help in producing yogurt cheese, hot sauce, pickles, fermented sausages, and kimchi. Molds are used to manufacture various food and food products. They help in ripening several types of food. Molds also help in producing enzymes and citric acid used in making bread and soft drinks respectively. Yeasts are used in the food industry as they ferment sugar into ethanol and carbon dioxide. A yeast, Saccharomyces cerevisiae is used to make bread, brew beer, and make wine. Nisin is an antimicrobial agent derived from bacteria. It is used in cheese, meats, and beverages to preserve them by inhibiting the growth of detrimental microorganisms. 2. Importance of Microbiology in Medical Science. Microorganisms can cause both benefit and harm to our human and animal cells. These microorganisms include virus, bacteria, fungus, and parasites. Medical Microbiology is important for several reasons. Through the knowledge of medical microbiology, microbiologists can identify, isolate, diagnose, prevent pathogenic microorganisms. They also can engineer beneficial microorganisms to produce antimicrobial drugs. The knowledge of microbiology helps microbiologists to innovate new methods to combat diseases. Fluorescent fusion is a good example of medical microbiology, that helps in the rapid detection of pathogens in the tissue sample. 3. Importance of Microbiology in the pharmaceutical industry. The discovery of antibiotics is one of the most important contributions of microbiology in the pharmaceutical industry. Antibiotics are the metabolic byproduct of microorganisms. The vaccine is also another important discovery of microbiology. The vaccine helps thousands of people by producing antibodies against the virus. Thus it prevents viral infection. For example, the Polio vaccine helps eradicate polio in many countries. Phage therapy has also attracted great attention due to its killing potential against bacteria. Steroids are also another pharmaceutical product produced by microorganisms. Prevention of microbial contamination of drugs, injectable, eye drops, nasal solutions, and inhalation products are also another important discoveries of microbiology. One of the great examples of medical microbiology is the discovery of Insulin. Insulin is used for the treatment of diabetic patients. Insulin was derived from the ß cell of pancreases. But it was not sufficient to satisfy the massive- demand for insulin. Recombinant DNA technology uses E. coli to produce a higher amount of insulin to control sugar concentration in diabetes patients. 4. Importance of Microbiology in nursing The knowledge of microbiology in nursing is very important to control and prevent infection in the hospital. The knowledge of microbiology for the nurses and other health professionals in healthcare is very important because it gives them much information about health and hygiene. Through the knowledge of microbiology, the nurses and other health professionals in healthcare can learn, how infections spread and how to carefully cure or surgery an open wound without infecting it. The knowledge of microbiology teaches them to keep the instruments aseptic and contaminant-free. It helps them find and identify the symptoms of an infection and type of infection at its early stage. It also teaches them the nature of the organism and the factors affecting its growth, the most susceptible means of disease transmission and the composition of chemicals, drugs, aseptic solutions, etc. 5. Importance of Microbiology in Biotechnology Microorganisms are used in various fields of biotechnology. In the fermentation industry, microbes are used to produce ethanol, organic acids, vinegar, and fermented foods by degrading complex organic matters. Microbes (e. g. virus) are used as a source of molecular vectors such as a plasmid, phagemid, and cosmid in molecular biology and recombinant DNA technology. Microbes are used in bioremediation that removes organic compounds and hydrocarbon from sewage water by degrading these organic wastes. By using microorganisms, microbiologists extract metals or heavy metals from their ore through a process called bioleaching and biomining. Microorganisms also produce enzymes, vitamins, organic acids, antibiotics, amino acids, and polysaccharides as their metabolic products for industrial purposes. 6. Importance of Microbiology in Chemical products Several industrial chemical products are produced through the use of microorganisms. These products include acetaldehyde, acetic acid, acetoacetic acid, butanol, ethanol, fructose, galactose, glycerol, lactic acid, mannitol, mannose, pyruvic acid, sorbose, succinic acid. The microorganism that produces these products include Acetic acid bacteria, lactic acid bacteria, propionic acid bacteria, butyric acid bacteria, E. coli, Aerobacter aerogenes. 7. Importance of Microbiology in fuel: Biofuel can be obtained from extracted microalgae oil, biomethane by anaerobic digestion of Microalgae biomass, ethanol by fermentation of carbohydrate. Biodiesel Microalgae are a potential source of oil content for biodiesel production. They also contain a higher amount of lipids that serve as raw material for biodiesel production. Butanol and ethanol Microalgae contain a large amount of cellulose, starch, mannitol, agar and laminarin that are fermented to alcohol (ethanol and butanol). This microalga includes Chlorella, Chlamydomonas, Dunaliella, Scenedesmus, and Spirulina. 8. Importance of Microbiology in aquaculture/fisheries: Heterotrophic microorganisms play an important role in decomposing organic matter and cycling of nutrients in aquatic systems. In the aquatic system, microbes are placed at the bottom of the food chain. Microorganisms also play various important roles in aquaculture as they grow naturally in the aquatic environment. Microorganisms may 1. Protect fish and larvae from various microbial infections. 2. Improve the overall quality of water.  3. Control the development of microbial and insect-infected diseases. 4. Suppress the growth of harmful bacteria and enhance larval survival, minimizing the need to apply antibiotics. 9. Importance of Microbiology in Agriculture: Microorganisms help in decomposing toxic compounds in agricultural soil preventing toxic accumulation in the soil. Thus it helps in increasing the fertility of the soil. Microorganisms (e.g. Blue-green algae) play an important role in nitrogen fixation. Fungi like Mycorrhiza help the plant absorb minerals and water from the soil and protect its roots from other fungi and nematodes. Actinomycetes (Nocardia and Monospora) and Molds (Mucor and Aspergillus) increase the fertility of the soil by decomposing its organic matter. Other soil bacteria help in nitrification, nitrification, mineralization and sulfur oxidation, etc. Soil bacteria produce Humus that helps in retaining soil moisture and enhances the formation of soil structure. Soil bacteria support plant growth by producing several substances like auxins, gibberellins, and antibiotics. Soil bacteria also help in controlling pest and microbial disease. A best-known soil bacterium Bacillus thuringienis have the ability to control caterpillar pests of plants. 10. Importance of Microbiology in Environmental Science/ Sewage System: Environmental Microbiology is the study of microorganisms that lives in air, soil, water. Microorganisms degrade toxic pollutants from the environment. For example, Acinetobacter can degrade a wide range of aromatic compounds. Bacteria help us clean water through sewage treatment. Bacteria degrade the organic matter in sewage removing the pollution from water. Nitrification and phosphorous removal are occurred by bacteria in the sewage system. In this system, Nitrosomonas spp oxidize ammonium to nitrite and Nitrobacter spp oxidize nitrite to nitrate. Denitrifying bacteria (Pseudomonas) reduce nitrate into nitrogen gas using the chemical energy in organic matter to reduce. Aerobic bacteria also ferment solid components of the sewage. Historical background Microbiology essentially began with the development of the microscope. Although others may have seen microbes before him, it was Antonie van Leeuwenhoek, a Dutch draper whose hobby was lens grinding and making microscopes, who was the first to provide proper documentation of his observations. His descriptions and drawings included protozoans from the guts of animals and bacteria from teeth scrapings. His records were excellent because he produced magnifying lenses of exceptional quality. Leeuwenhoek conveyed his findings in a series of letters to the British Royal Society during the mid-1670s. Although his observations stimulated much interest, no one made a serious attempt either to repeat or to extend them. Leeuwenhoek's "animalcules," as he called them, thus remained mere oddities of nature to the scientists of his day, and enthusiasm for the study of microbes grew slowly. It was only later, during the 18th-century revival of a long-standing controversy about whether life could develop out of nonliving material, that the significance of microorganisms in the scheme of nature and in the health and welfare of humans became evident. Spontaneous generation versus biotic generation of life The early Greeks believed that living things could originate from nonliving matter (abiogenesis) and that the goddess Gea could create life from stones. Aristotle discarded this notion, but he still held that animals could arise spontaneously from dissimilar organisms or from soil. His influence regarding this concept of spontaneous generation was still felt as late as the 17th century, but toward the end of that century a chain of observations, experiments, and arguments began that eventually refuted the idea. This advance in understanding was hard fought, involving series of events, with forces of personality and individual will often obscuring the facts. Although Francesco Redi, an Italian physician, disproved in 1668 that higher forms of life could originate spontaneously, proponents of the concept claimed that microbes were different and did indeed arise in this way. Such illustrious names as John Needham and Lazzaro Spallanzani were adversaries in this debate during the mid-1700s. In the early half of the 1800s, Franz Schulze and Theodor Schwann were major figures in the attempt to disprove theories of abiogenesis until Louis Pasteur finally announced the results of his conclusive experiments in 1864. In a series of masterful experiments, Pasteur proved that only preexisting microbes could give rise to other microbes (biogenesis). Modern and accurate knowledge of the forms of bacteria can be attributed to German botanist Ferdinand Cohn, whose chief results were published between 1853 and 1892. Cohn's classification of bacteria, published in 1872 and extended in 1875, dominated the study of these organisms thereafter. Microbes and disease Girolamo Fracastoro, an Italian scholar, advanced the notion as early as the mid-1500s that contagion is an infection that passes from one thing to another. A description of precisely what is passed along eluded discovery until the late 1800s, when the work of many scientists, Pasteur foremost among them, determined the role of bacteria in fermentation and disease. Robert Koch, a German physician, defined the procedure (Koch's postulates) for proving that a specific organism causes a specific disease. The foundation of microbiology was securely laid during the period from about 1880 to 1900. Students of Pasteur, Koch, and others discovered in rapid succession a host of bacteria capable of causing specific diseases (pathogens). They also elaborated an extensive arsenal of techniques and laboratory procedures for revealing the ubiquity, diversity, and abilities of microbes. Progress in the 20th Century All of these developments occurred in Europe. Not until the early 1900s did microbiology become established in America. Many microbiologists who worked in America at this time had studied either under Koch or at the Pasteur Institute in Paris. Once established in America, microbiology flourished, especially with regard to such related disciplines as biochemistry and genetics. In 1923 American bacteriologist David onercan Bacerbgs Davd Bergey established that science's primary reference, updated editions of which continue to be used today. Since the 1940s microbiology has experienced an extremely productive period during which many disease-causing microbes have been identified and methods to control them developed. Microorganisms have also been effectively utilized in industry; their activities have been channeled to the extent that valuable products are now both vital and commonplace. The study of microorganisms has also advanced the knowledge of all living things. Microbes are easy to work with and thus provide a simple vehicle for studying the complex processes of life; as such they have become a powerful tool for studies in genetics and metabolism at the molecular level. This intensive probing into the functions of microbes has resulted in numerous and often unexpected dividends. Knowledge of the basic metabolism and nutritional requirements of a pathogen, for example, often leads to a means of controlling disease or infection. Types of microorganisms The major groups of microorganisms-namely bacteria, archaea, fungi (yeasts and molds), algae, protozoa, and viruses-are summarized below. Bacteria (eubacteria and archaea) Microbiology came into being largely through studies of bacteria. The experiments of Louis Pasteur in France, Robert Koch in Germany, and others in the late 1800s established the importance of microbes to humans. As stated in the Historical background section, the research of these scientists provided proof for the germ theory of disease and the germ theory of fermentation. It was in their laboratories that techniques were devised for the microscopic examination of specimens, culturing (growing) microbes in the laboratory, isolating pure cultures from mixed-culture populations, and many other laboratory manipulations. These techniques, originally used for studying bacteria, have been modified for the study of all microorganisms—hence the transition from bacteriology to microbiology. The organisms that constitute the microbial world are characterized as either prokaryotes or eukaryotes; all bacteria are prokaryotic-that is, single-celled organisms without a membrane-bound nucleus. Their DNA (the genetic material of the cell), instead of being contained in the nucleus, exists as a long, folded thread with no specific location within the cell. Until the late 1970s it was generally accepted that all bacteria are closely related in evolutionary development. This concept was challenged in 1977 by Carl R. Woese and coinvestigators at the University of Illinois, whose research on ribosomal RNA from a broad spectrum of living organisms established that two groups of bacteria evolved by separate pathways from a common and ancient ancestral form. This discovery resulted in the establishment of a new terminology to identify the major distinct groups of microbes-namely, the eubacteria (the traditional or "true" bacteria), the archaea (bacteria that diverged from other bacteria at an early stage of evolution and are distinct from the eubacteria), and the eukarya (the eukaryotes). Today the eubacteria are known simply as the true bacteria (or the bacteria) and form the domain Bacteria. The evolutionary relationships between various members of these three groups, however, have become uncertain, as comparisons between the DNA sequences of various microbes have revealed many puzzling similarities. As a result, the precise ancestry of today's microbes is very difficult to resolve. Even traits thought to be characteristic of distinct taxonomic groups have unexpectedly been observed in other microbes. For example, an anaerobic ammonia-oxidizer—the "missing link" in the global nitrogen cycle was isolated for the first time in 1999. This bacterium (an aberrant member of the order Planctomycetales) was found to have internal structures similar to eukaryotes, a cell wall with archaean traits, and a form of reproduction (budding) similar to that of yeast cells. Bacterial Cell Bacteria have a variety of shapes, including spheres, rods, and spirals. Individual cells generally range in width from 0.5 to 5 micrometres (um; millionths of a metre). Although unicellular, bacteria often appear in pairs, chains, tetrads (groups of four), or clusters. Some have flagella, external whiplike structures that propel the organism through liquid media; some have capsule, an exteral coating of the cell; some produce spores— reproductive bodies that function much as seeds do among plants. One of the major characteristics of bacteria is their reaction to the Gram stain. Depending upon the chemical and structural composition of the cell wall, some bacteria are gram-positive, taking on the stain's purple colour, whereas others are gram-negative. Through a microscope the archaea look much like bacteria, but there are important differences in their chemical composition, biochemical activities, and environments. The cell walls of all true bacteria contain the chemical substance peptidoglycan, whereas the cell walls of archaeans lack this substance. Many archaeans are noted for their ability to survive unusually harsh surroundings, such as high levels of salt or acid or high temperatures. These microbes, called extremophiles, live in such places as salt flats, thermal pools, and deep-sea vents. Some are capable of a unique chemical activity— the production of methane gas from carbon dioxide and hydrogen. Methane- producing archaea live only in environments with no oxygen, such as swamp mud or the intestines of ruminants such as cattle and sheep. Collectively, this group of microorganisms exhibits tremendous diversity in the chemical changes that it brings to its environments. Algae The cells of eukaryotic microbes are similar to plant and animal cells in that their DNA is enclosed within a nuclear membrane, forming the nucleus. Eukaryotic microorganisms include algae, protozoa, and fungi. Collectively algae, protozoa, and some lower fungi are frequently referred to as protists (kingdom Protista, also called Protoctista); some are unicellular and others are multicellular. representative algae Unlike bacteria, algae are eukaryotes and, like plants, contain the green pigment chlorophyll, carry out photosynthesis, and have rigid cell walls. They normally occur in moist soil and aquatic environments. These eukaryotes may be unicellular and microscopic in size or multicellular and up to 120 metres (nearly 400 feet) in length. Algae as a group also exhibit a variety of shapes. Single-celled species may be spherical, rod-shaped, club-shaped, or spindle-shaped. Some are motile. Algae that are multicellular appear in a variety of forms and degrees of complexity. Some are organized as filaments of cells attached end to end; in some species these filaments intertwine into macroscopic, plantlike bodies. Algae also occur in colonies, some of which are simple aggregations of single cells, while others contain different cell types with special functions. Fungi Fungi are eukaryotic organisms that, like algae, have rigid cell walls and may be either unicellular or multicellular. Some may be microscopic in size, while others form much larger structures, such as mushrooms and bracket fungi that grow in soil or on damp logs. Unlike algae, fungi do not contain chlorophyll and thus cannot carry out photosynthesis. Fungi do not ingest food but must absorb dissolved nutrients from the environment. Of the fungi classified as microorganisms, those that are multicellular and produce filamentous, microscopic structures are frequently called molds, whereas yeasts are unicellular fungi. In molds cells are cylindrical in shape and are attached end to end to form threadlike filaments (hyphae) that may bear spores. Individually, hyphae are microscopic in size. However, when large numbers of hyphae accumulate for example, on a slice of bread or fruit jelly-they form a fuzzy mass called a mycelium that is visible to the naked eye. The unicellular yeasts have many forms, from spherical to egg-shaped to filamentous. Yeasts are noted for their ability to ferment carbohydrates, producing alcohol and carbon dioxide in products such as wine and bread. Protozoa representative protozoans Protozoa, or protozoans, are single-celled, eukaryotic microorganisms. Some protozoa are oval or spherical, others elongated. Still others have different shapes at different stages of the life cycle. Cells can be as small as 1 um in diameter and as large as 2,000 um, or 2 mm (visible without magnification). Like animal cells, protozoa lack cell walls, are able to move at some stage of their life cycle, and ingest particles of food; however, some phytoflagellate protozoa are plantlike, obtaining their energy via photosynthesis. Protozoan cells contain the typical internal structures of an animal cell. Some can swim through water by the beating action of short, hairlike appendages (cilia) or flagella. Their rapid, darting movement in a drop of pond water is evident when viewed through a microscope. amoeba The amoebas (also amoebae) do not swim, but they can creep along surfaces by extending a portion of themselves as a pseudopod and then allowing the rest of the cell to flow into this extension. This form of locomotion is called amoeboid movement. The sporozoans (phylum Apicomplexa) are so named because they form dormant bodies called spores during one phase of their life cycle. Protozoa occur widely in nature, particularly in aquatic environments. Viruses tobacco mosaic virus Viruses, agents considered on the borderline of living organisms, are also included in the science of microbiology, come in several shapes, and are widely distributed in nature, infecting animal cells, plant cells, and microorganisms. The field of study in which they are investigated is called virology. All viruses are obligate parasites; that is, they lack metabolic machinery of their own to generate energy or to synthesize proteins, so they depend on host cells to carry out these vital functions. Once inside a cell, viruses have genes for usurping the cell's energy-generating and protein- synthesizing systems. In addition to their intracellular form, viruses have an extracellular form that carries the viral nucleic acid from one host cell to another. In this infectious form, viruses are simply a central core of nucleic acid surrounded by a protein coat called a capsid. The capsid protects the genes outside the host cell; it also serves as a vehicle for entry into another host cell because it binds to receptors on cell surfaces. The structurally mature, infectious viral particle is called a virion. With the electron microscope it is possible to determine the morphological characteristics of viruses. Virions generally range in size from 20 to 300 nanometres (nm; billionths of a metre). Since most viruses measure less than 150 mm, they are beyond the limit of resolution of the light microscope and are visible only by electron microscopy. By using materials of known size for comparison, microscopists can determine the size and structure of individual virions. Prions Even smaller than viruses, prions (pronounced "pree-ons") are the simplest infectious agents. Like viruses they are obligate parasites, but they possess no genetic material. Although prions are merely self-perpetuating proteins, they have been implicated as the cause of various diseases, including bovine spongiform encephalopathy (mad cow disease), and are suspected of playing a role in a number of other disorders. Lichens crustose lichens Lichens represent a form of symbiosis, namely, an association of two different organisms wherein each benefits. A lichen consists of a photosynthetic microbe (an alga or a cyanobacterium) growing in an intimate association with a fungus. A simple lichen is made up of a top layer consisting of a tightly woven fungal mycelium, a middle layer where the photosynthetic microbe lives, and a bottom layer of mycelium. In this mutualistic association, the photosynthetic microbes synthesize nutrients for the fungus, and in return the fungus provides protective cover for the algae or cyanobacteria. Lichens play an important role ecologically; among other activities they are capable of transforming rock to soil. Slime molds The slime molds are a biological and taxonomic enigma because they are neither typical fungi nor typical protozoa. During one of their growth stages, they are protozoa-like because they lack cell walls, have amoeboid movement, and ingest particulate nutrients. During their propagative stage they form fruiting bodies and sporangia, which bear walled spores like typical fungi. Traditionally, the slime molds have been classified with the fungi. There are two groups of slime molds: the cellular slime molds and the acellular slime molds. The study of microorganisms As is the case in many sciences, the study of microorganisms can be divided into two generalized and sometimes overlapping categories. Whereas basic microbiology addresses questions regarding the biology of microorganisms, applied microbiology refers to the use of microorganisms to accomplish specific objectives. Basic microbiology The study of the biology of microorganisms requires the use of many different procedures as well as special equipment. The biological characteristics of microorganisms can be summarized under the following categories: morphology, nutrition, physiology, reproduction and growth, metabolism, pathogenesis, antigenicity, and genetic properties. Morphology Morphology refers to the size, shape, and arrangement of cells. The observation of microbial cells requires not only the use of microscopes but also the preparation of the cells in a manner appropriate for the particular kind of microscopy. During the first decades of the 20th century, the compound light microscope was the instrument commonly used in microbiology. Light microscopes have a usual magnification factor of 1000 x and a maximum useful magnification of approximately 2000 x. Specimens can be observed either after they have been stained by one of several techniques to highlight some morphological characteristics or in living, unstained preparations as a "wet mount." Light microscopy Several modifications of light microscopy are available, such as: bright field The specimen is usually stained and observed while illuminated; useful for observation of the gross morphological features of bacteria, fungi, algae, and protozoa. dark field The specimen is suspended in a liquid on a special slide and can be observed in a living condition; useful for determining motility of microorganisms or some special morphological characteristic such as spiral or coiled shapes. fluorescence The specimen is stained with a fluorescent dye and then illuminated; objects that take up the fluorescent dye will "glow." phase contrast Special condenser lenses allow observation of living cells and differentiation of cellular structures of varying density. Electron microscopy The development of the electron microscope and complimentary techniques vastly increased the resolving power beyond that attainable with light microscopy. This increase is possible because the wavelengths of the electron beams are so much shorter than the wavelengths of light. Objects as small as 0.02 m are resolvable by electron microscopy, compared with 0.25 um- allowing, for instance, the observation of virions and viral structures. Specimens are observed by either transmission electron microscopy or scanning electron microscopy. In TEM the electron beam passes through the specimen and registers on a screen forming the image; in SEM the electron beam moves back and forth over the surface of microorganisms coated with a thin film of metal and registers a three-dimensional picture on the screen. Advances in microscopes and microscopic techniques continue to be introduced to study cells, molecules, and even atoms. Among these are confocal microscopy, the atomic force microscope, the scanning tunneling microscope, and immunoelectron microscopy. These are particularly significant for studies of microorganisms at the molecular level. Nutritional and physiological characteristics Microorganisms as a group exhibit great diversity in their nutritional requirements and in the environmental conditions that will support their growth. No other group of living organisms comes close to matching the versatility and diversity of microbes in this respect. Some species will grow in a solution composed only of inorganic salts (one of the salts must be a compound of nitrogen) and a source of carbon dioxide (COz); these are called autotrophs. Many, but not all, of these microbes are autotrophic via photosynthesis. Organisms requiring any other carbon source are called heterotrophs. These microbes commonly make use of carbohydrates, lipids, and proteins, although many microbes can metabolize other organic compounds such as hydrocarbons. Others, particularly the fungi, are decomposers. Many species of bacteria also require specific additional nutrients such as minerals, amino acids, and vitamins. Various protozoans, fungi, and bacteria are parasites, either exclusively (obligate parasites) or with the ability to live independently (facultative parasites). If the nutritional requirements of a microorganism are known, a chemically defined medium containing only those chemicals can be prepared. More complex media are also routinely used; these generally consist of peptone (a partially digested protein), meat extract, and sometimes yeast extract. When a solid medium is desired, agar is added to the above ingredients. Agar is a complex polysaccharide extracted from marine algae. It has several properties that make it an ideal solidifying substance for microbiological media, particularly its resistance to microbial degradation. Microorganisms vary widely in terms of the physical conditions required for growth. For example, some are aerobes (require oxygen), some are anaerobes (grow only in the absence of oxygen), and some are facultative (they grow in either condition). Eukaryotic microbes are generally aerobic. Microorganisms that grow at temperatures below 20 °C (68 °F) are called psychrophiles; those that grow best at 20-40 °C (68-104 °F) are called mesophiles; a third group, the thermophiles, require temperatures above 40 °C. Those organisms which grow under optimally under one or more physical or chemical extremes, such as temperature, pressure, pH, or salinity, are referred to as extremophiles. Bacteria exhibit the widest range of temperature requirements. Whereas bacterial (and fungal) growth is commonly observed in food that has been refrigerated for a long period, some isolated archaea (e.g., Pyrodictium occultum and Pyrococcus woesel) grow at temperatures above 100 °C (212 °F). Other physical conditions that affect the growth of microorganisms are acidity or basicity (pH), osmotic pressure, and hydrostatic pressure. The optimal pH for most bacteria associated with the human environment is in the neutral range near pH 7, though other species grow under extremely basic or acidic conditions. Most fungi are favoured by a slightly lower pH (5-6); protozoa require a range of pH 6.7-7.7; algae are similar to bacteria in their requirements except for the fact that they are photosynthetic. Reproduction and growth Bacteria reproduce primarily by binary fission, an asexual process whereby a single cell divides into two. Under ideal conditions some bacterial species may divide every 10-15 minutes-a doubling of the population at these time intervals. Eukaryotic microorganisms reproduce by a variety of processes, both asexual and sexual. Some require multiple hosts or carriers (vectors) to complete their life cycles. Viruses, on the other hand, are produced by the host cell that they infect but are not capable of self-reproduction. The study of the growth and reproduction of microorganisms requires techniques for cultivating them in pure culture in the laboratory. Data collected on the microbial population over a period of time, under controlled laboratory conditions, allow a characteristic growth curve to be constructed for a species. Metabolism Collectively, microorganisms show remarkable diversity in their ability to produce complex substances from simple chemicals and to decompose complex materials to simple chemicals. An example of their synthetic ability is nitrogen fixation-the production of amino acids, proteins, and other organic nitrogen compounds from atmospheric nitrogen (N2). Certain bacteria and blue-green algae (cyanobacteria) are the only organisms capable of this ecologically vital process. An example of microbes' ability to decompose complex materials is shown by the white and brown rot fungi that decompose wood to simple compounds, including COz. Laboratory procedures are available that make it possible to determine the biochemical capability of a species qualitatively and quantitatively. Routine techniques can identify which compounds or substances are degraded by a specific microbe and which products are synthesized. Through more elaborate experimentation it is possible to determine step-by-step how the microbe performs these biochemical changes. Studies can be performed in a number of ways using growing cultures, "resting cells" (suspensions of cells), cell-free extracts, or enzyme preparations from cells. Certain biochemical tests are routinely used to identify microbes-though more in the case of bacteria than algae, fungi, or protozoa. The adoption of routine sets of laboratory tests has allowed automated instrumentation to perform the tests. For instance, technicians often simply inoculate individual units of a "chamber" that is preloaded with a specific chemical substance (the substrate) and then place the chamber into an apparatus that serves as an incubator and analyzer. The apparatus automatically records the results and is frequently capable of calculating the degree of accuracy of the identification. Pathogenesis Some microorganisms cause diseases of humans, other animals, and plants. Such microbes are called pathogens. Pathogens are identified by the hosts they infect and the symptoms they cause; it is also important to identify the specific properties of the pathogen that contribute to its infectious capacity— a characteristic known as virulence. The more virulent a pathogen, the fewer the number needed to establish an infection. Antigenic characteristics An antigen is a substance that, when introduced into an animal body, stimulates the production of specific substances (antibodies) that react or unite with the antigen. Microbial cells and viruses contain a variety of antigenic substances. A significant feature of antigen-antibody reactions is specificity; the antibodies formed as a result of inoculating an animal with one microbe will not react with the antibodies formed by inoculation with a different microbe. Antibodies appear in the blood serum of animals, and laboratory tests of antigen-antibody reactions are performed by using sera-hence the term serological reactions. Thus, it is possible to characterize a microorganism by its antigenic makeup as well as to identify microorganisms by using one of many different serological tests. Antigens and antibodies are important aspects of immunity, and immunology is included in the science of microbiology. Genetic characterization Since the last quarter of the 20th century, researchers have accumulated a vast amount of information elucidating in precise detail the chemical composition, synthesis, and replication of the genetic material of cells. Much of this research has been done by using microorganisms, and techniques have been developed that permit experimentation at the molecular level. For instance, experiments determining the degree of similarity between different organisms' DNA and RNA have provided new insights for the classification of microorganisms. Test kits are available for the identification of microorganisms, particularly bacteria, by DNA probes. Since the invention of recombinant DNA technology in 1973, techniques have been developed whereby genes from one cell can be transferred to an entirely different cell, as when a gene is transferred from an animal cell to a bacterium or from a bacterium to a plant cell. Recombinant DNA technology opened the door to many new medical and industrial applications of microbiology, and it plays a central role in genetic engineering. Applied microbiology Genetic engineering is an example of how the fields of basic and applied microbiology can overlap. Genetic engineering is primarily considered a field of applied microbiology (that is, the exploitation of microorganisms for a specific product or use). The methods used in genetic engineering were developed in basic research of microbial genetics. Conversely, methods used and perfected for applied microbiology can become tools for basic microbiology. Applied microbiology can, however, be divided under the following headings. Soil microbiology However "dead" soil may appear, it is in fact teeming with millions or billions of microbial cells per gram, depending upon soil fertility and the environment. Dead vegetation, human and animal wastes, and dead animals are deposited in or on soil. In time they all decompose into substances that contribute to soil, and microbes are largely responsible for these transformations. Two great pioneer soil microbiologists were Martinus W. Beijerinck (1851- 1931), a Dutchman, and Sergey N. Winogradsky (1856-1953), a Russian. These researchers isolated and identified new types of bacteria from soil, particularly autotrophic bacteria, that use inorganic chemicals as nutrients and as a source of energy. The relationship between legumes and bacteria in the nodules of legume roots was discovered by other scientists in 1888. The nodules contain large numbers of bacteria (Rhizobium) that are capable of fixing atmospheric nitrogen into compounds that can be used by plants. The ecology of fertile soil consists of plant roots, animals such as rodents, insects, and worms, and a menagerie of microorganisms-viruses, bacteria, algae, fungi, and protozoa. The role of this microbial flora can be conveniently expressed in Earth's natural cycles. In the nitrogen cycle, for example, microorganisms capture nitrogen gas from the atmosphere and convert it into a combined form of nitrogen that plants can use as a nutrient; the plant synthesizes organic nitrogen compounds that are consumed by humans and animals; the consumed nitrogen compounds eventually reach the soil; microorganisms complete the cycle by decomposing these compounds back to atmospheric nitrogen and simple inorganic molecules that can be used by plants. In similar cycles for other elements such as carbon, sulfur, and phosphorus, microbes play a role; this makes them essential to maintaining life on Earth. Microbiology of water supplies, wastewater, and other aquatic environments Long before the establishment of microbiology as a science, water was suspected of being a carrier of disease-producing organisms. But it was not until 1854, when an epidemic of cholera was proved to have had its origin in polluted water, that contaminated water was considered more seriously as a source of disease. Since that time there has been continuous research on the microbiology of public water supplies, including the development of laboratory procedures to determine whether the water is potable, or safe for human consumption. At the same time, purification procedures for these supplies have emerged. A highly standardized and routine laboratory procedure to determine the potability of water is based upon detecting the presence or absence of the bacterium Escherichia coli. E. coli is a normal inhabitant of the intestinal tract of humans; its presence in water indicates that the water is polluted with intestinal wastes and may contain disease-producing organisms. The principal operations employed in a municipal water-purification plant are sedimentation, filtration, and chlorination. Each of these operations removes or kills microorganisms, and the microbiological quality of the treated water is monitored at frequent intervals. The used water supply of a community, commonly referred to as sewage, is microbiologically significant in two ways. First, sewage is a potential carrier of pathogenic microorganisms, so measures such as chlorination must be implemented to prevent these microbes from contaminating drinking-water supplies. Second, sewage-treatment plants purify water by exploiting the biochemical abilities of microbes to metabolize contaminants. Raw sewage is processed through large tanks, first for anaerobic degradation of complex substrates and later for aerobic oxidation of soluble products. This "activated sludge" treatment is dependent upon incubation conditions that favour the growth and metabolic activity of appropriate microorganisms. Another aspect of the microbiology of water pertains to natural bodies of water such as ponds, lakes, rivers, and oceans. Aquatic microbes perform a host of biochemical transformations and are an essential component of the food chain in these environments. For example, the microbial flora of the sea comprises bacteria, algae, fungi, and protozoa. The microorganisms inhabiting aquatic environments are collectively referred to as plankton; phytoplankton refers to the photosynthetic microbes (primarily algae), whereas protozoa, and other small animals, are zooplankton. Phytoplankton is responsible for converting solar energy into chemical energy-the components of plankton cells that serve as food for higher aquatic life. The magnitude of this process can be appreciated by calculations indicating that it takes 1,000 tons of phytoplankton to support the growth of one ton of fish. Large populations of archaea live in volcanic ridges 2,600 metres (8,500 feet) below the ocean surface in areas immediately surrounding hydrothermal vents (deep-sea hot springs). The vents spew superheated water (350 °C [662 °F]) that contains hydrogen sulfide (HS); the water surrounding the vents has a temperature range of 10-20 °C (50-68 °F). Many bacteria concentrate in this region because of the availability of H2S, which they can use for energy. The abundance of animal life that also inhabits this region is completely dependent on the microbes for food. There is a growing interest in other ecological aspects of aquatic microbiology, such as the role of microbes in global warming and oxygen production. Experimental approaches are being developed to study the complex biology and ecology of biofilms and microbial mats. These assemblages of microbes and their products, while potentially useful in several ways, are complex. In many instances the microbial flora involved must sometimes be studied in its natural environment because the environment cannot be reproduced in the laboratory. Food microbiology Microorganisms are of great significance to foods for the following reasons: (1) microorganisms can cause spoilage of foods, (2) microorganisms are used to manufacture a wide variety of food products, and (3) microbial diseases can be transmitted by foods. Food spoilage Foods can be considered as a medium for microbial growth. Considering the vast array of sources, substances, and methods with which food is produced, practically every kind of microbe is a potential contaminant. Given a chance to grow, microbes will produce changes in appearance, flavour, odour, and other qualities of the food. The changes vary according to the type of food degraded but can be summarized by examining the fates of the major nutrients found in food: proteins, carbohydrates, and fats. Protein-containing foods, particularly meats, are putrefied by organisms (e.g., Proteus, Pseudomonas, and Clostridium bacteria) that break down the long peptide chains of proteins into amino acids and foul-smelling compounds such as amines, ammonia, and hydrogen sulfide (HS). Carbohydrates (sugars and starches) are fermented into acids (e.g., the acetic acid in vinegar), alcohols, and gases, especially carbon dioxide. This process is responsible for the bursting of spoiled chocolate cream candies by yeasts. Fat-containing foods such as dairy products are spoiled by microbes that break down lipids into fatty acids and glycerol. Rancid milk, which can be caused by bacteria, yeast, or mold, is an example of this process. Improperly canned foods are also subject to spoilage by bacteria, yeasts, and molds. Bacteria such as Bacillus and Clostridium are of particular significance in the canning industry because of the high level of resistance that their spores possess. One example of microbial spoilage of canned foods is "sulfide spoilage" caused by C. nigrificans, in which contents are blackened and have the odour of rotten eggs. Another example is called "flat sour," in which the spoiled product has an abnormal odour, a cloudy appearance, and a sour taste owing to its lowered pH. Putrefaction caused by C. sporogenes may cause a can to swell and burst, releasing its partially digested contents and a putrid odour. Food preservation All methods of food preservation are based upon one or more of the following principles: (1) prevention of contamination and removal of microorganisms, (2) inhibition of microbial growth and metabolism, and (3) killing of microorganisms. Prevention-or, more accurately, minimization-of contamination is achieved by the sanitary handling of raw food products, inhibition of growth by low temperatures (refrigeration or freezing), dehydration by evaporation or by high concentrations of salt or sugar, and killing of microbes by the application of high temperatures and, in some instances, radiation. Food products from microorganisms Important food items produced in whole or in part by the biochemical activities of microorganisms include pickles, sauerkraut, olives, soy sauce, certain types of sausage, all unprocessed cheeses except cream cheese, and many fermented milk products such as yogurt and acidophilus milk. In each instance a raw food item, such as cucumbers in the case of pickles or milk protein in the case of cheeses, is inoculated with microorganisms known to produce the changes required for a desirable product. The initial food item thus serves as a substrate that is acted upon by microorganisms during the period of incubation. Frequently the manufacturer uses a "starter culture"—a commercial population of microorganisms already known to produce a good product. Industrial microbiology and genetic engineering Many substances of considerable economic value are products of microbial metabolism. From an industrial viewpoint the substrate may be regarded as a raw material and the microorganism as the "chemical factory" for converting the raw material into new products. If an organism can be shown to convert inexpensive raw material into a useful product, it may be feasible to perform this reaction on a large industrial scale if the following conditions can be met. The organism. The organism to be employed (a virus, bacterium, yeast, or mold) must have the capacity to produce appreciable amounts of the product. It should have relatively stable characteristics and the. ability to grow rapidly and vigorously, and it should be nonpathogenic. The medium. The medium, including the substrate from which the organism produces the new product, must be cheap and readily available in large quantities. The product. A feasible method of recovering and purifying the desired end product must be developed. Industrial fermentations are performed in large  tanks, some with capacities of 190,000 litres (50,000 gallons) or more. The product formed by the metabolism of the microorganism must be removed from a heterogeneous mixture that also includes a tremendous crop of microbial cells and unused constituents of the medium, as well as products of metabolism other than those being sought. Traditional products of industrial microbiology are antibiotics, alcoholic beverages, vaccines, vinegar, and miscellaneous chemicals such as acetone and butyl alcohol. The development of recombinant DNA technology, however, has made it possible to conceive of virtually unlimited new products made by genetically engineered microorganisms. One example of what can be achieved via recombinant DNA technology is the production of human insulin by a genetically altered strain of E. coli. By inserting the human gene coding for insulin into the E. coli cell, biotechnologists give this bacterium the ability to synthesize the hormone on an industrial scale. The scientific advances that have made genetic engineering a reality have broad implications for the future. By introducing foreign genes into microorganisms, it may be possible to develop strains of microbes that offer new solutions to such diverse problems as pollution, food and energy shortages, and the treatment and control of disease.