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Microbiology is the study of living organisms of microscopic size. The term microbiology was given by French chemist Louis Pasteur (1822-95). The word ‗Microbiology‘ is derived from three Greek wordsMikros (small), Bios (living) & Logos (science). Microbiology is said to have its roots in the great expansion and development of the biological sciences that took place after 1850. The term microbe was first used by Sedillot (1878). Microorganisms like all other living organisms possess the following characters: 1. 2. 3. 4. 5. Ability to reproduce Ability to ingest food materials Ability to excrete waste products Irritability Susceptibility to mutation
The discovery of microbiology as a discipline could be traced along the following historical eras:
In 20th Century: Era of Molecular Biology
This period concerns with the discovery of microbial world that has been dominated by Antony Van Leeuwenhoek.
Concepts of Aristotle: Aristotle (384 – 322 B.C.) believed that the animals might originate
spontaneously from the soil, plants, or other unlike animals. His influence was still strongly felt in the seventeenth century!!
Antony Van Leeuwenhoek (1632-1723):
Of Delft, Holland (Netherland) he was the first person to observe and accurately describe microorganisms (bacteria and protozoa) called 'animalcules' (little animals) in 1676. Actually he was a Dutch linen merchant but spent much of his spare time constructing simple microscopes composed of double convex lenses held between two silver plates. He constructed over 250 small powerful microscopes that could magnify around 50-300 times. Leeuwenhoek was the 1st person to produce precise and correct descriptions of bacteria and protozoa using microscope he made himself. Because of this extraordinary contribution to microbiology he is considered as the "Father of bacteriology and protozoology". He wrote over 200 letters which was transmitted as a series of letters from 1674-1723 to Royal society in London during a 50 years period.
Spontaneous Generation and Biogenesis:
Early belief that some forms of life could arise from ―vital forces‖ present in nonliving or decomposing matter, abiogenesis. In other words, organisms can arise from non-living matter. The alternative hypothesis, that the living organisms arise from preexisting life, is called biogenesis.
Lazzaro Spallanzani (1729-1799): He was an Italian Naturalist who attempted to refute Needham's experiment. Nicolas Appert followed the idea of Spallanzani's work. John Needham (1713-1781): He was probably the greatest supporter of the theory of spontaneous generation. Experiment of Franz Schulze and Theodor Schwann: Two German scholars Schulze (1815-1873) and Theodor Schwann (1810-1882) viewed that air was the source of microbes and sought to prove this by passing air through hot glass tubes or strong chemicals into boiled infusions in flasks. he boiled beef broth for longer period. Followed incubation no growth was observed by him in these flasks. Louis Jablot (1670): In 1670. removed the air from the flask and then sealed the container. Redi was the first who put the theory of spontaneous generation to test by conducting a simple experiment in which he placed meat in three jars. . The infusion in both the cases remained free from the microbes. while experimenting with raw meat exposed to hot ashes. He proposed that tiny organisms the animalcules arose spontaneously on his mutton gravy. But he failed to convince Needham. left uncovered and landed on meat where they laid their egg that later developed into maggots. second was covered with paper and third was left uncovered. an Italian physician. Flies entered the jar that was open to air i. However flies were attracted to the gauze covered jars and laid their eggs on the gauze and maggots subsequently developed without access to the meat. In 1749. John Needham. The other two pieces of meat did not produce maggots spontaneously. They said. indicating that maggots were the offspring of the flies and did not arise from some 'vital source' in the meat as previously believed. Thus Spallanzani disproved the doctrine of spontaneous generation. Francesco Redi (1626-1697): The ancient belief in spontaneous generation was first of all challenged by Redi. there were a number of significant developments in microbiology during Van Leeuwenhoek’s time. But the die-hard advocates of spontaneous generation were still not convinced. One jar was covered with fine gauze.TRANSITION PERIOD Although. Ignaz Semmelweis and John Snow were the two persons who showed a growing awareness of the mode of disease transmission. He was a French wine maker who showed that soups and liquids can be preserved by heating them extensively in thick champagnes bottles. Lazzaro Spallanzani and Nicolas Appert etc and to know the disease transmission which mainly includes the work of Ignaz Semmelweis and John Snow. The main aspects were to solve the controversy over spontaneous generation which includes experimentations mainly of Francesco Redi. John Needham observed the appearance of organisms not present at the start of the experiment and concluded that the bacteria virtually originated from the raw meat itself. Jablot conducted an experiment in which he divided a hay infusion that had been boiled into two containers: a heated container that was closed to the air and a heated container that was freely open to the air. He proposed this observation as an example of spontaneous generation. who carried out a series of experiments on decaying meat and its ability to produce maggots spontaneously. Only the open vessel developed microorganisms. who insisted that air was essential to the spontaneous production of microscopic beings and that it had been excluded from the flasks by sealing them. He showed that the heated nutrients could still grow animalcules when exposed to air by simply making a small crack in the neck.e. In 1749. acid and heat altered the air so that it would not support growth. Peoples were interested to correlate diseases with microbes. This further helped to disprove abiogenesis.
John Tyndall (1820 . and characterized ‗certain microbes‘ exclusively responsible for the ‗good batches‘ predominantly in comparison to the ones found solely in the ‗poor products‘. He conducted experiments in an aseptically designed box to prove that dust indeed carried the germs. typified. France. He demonstrated that if no dust was present. sterile broth remained free of microbial growth for indefinite period even if it was directly exposed to air. France manufactured large variety of wines and beer. later known as endospore. and thus. but just sufficient to cause and afford the legitimate destruction of a relatively very high . Fermentation and Pasteurization: Louis Pasteur began his brilliant career as professor of chemistry at the University of Lille. it ultimately initiated and gave rise to a basic technique of ‗plugging‘ bacterial culture tubes with ‗cotton plugs‘ (stoppers). He filled several round bottomed flasks with nutrient solution and fashioned their openings into elongated. microbial growth commenced immediately. In fact. Prolonged boiling or intermittent heating was necessary to kill these spores. a French microbiologist. Pasteur explored and exploited the unique capabilities of microbes in the fermentation industry exclusively using fruits and grains resulting in alcohol-based table wines. performed a series of experiments to prove that microorganisms were present in the air and were not spontaneously produced. Schroder and T.1893): An English physicist. rather than any magic. He was of the opinion that disease may be a biological phenomenon. He discovered highly resistant bacterial structure. This experiment clearly showed that microorganisms present on or in non-living materials such as dust or water were responsible for the contamination of sterile solutions. At that time as a principal industry. in the infusion of hay. von Dusch: George Schroeder and Theodor Von Dusch (1854) were the first to introduce the idea of using cotton plugs for plugging microbial culture tubes. to make the infusion completely sterilized. deal a final blow to spontaneous generation in 1877. the overall net outcome of such extensive as well as intensive investigations helped in a long way for the assured and successful production of consistently good and uniform ultimate product. Louis Pasteur: Pasteur. Pasteur meticulously isolated. But the concept of spontaneous generation was revived for the last time by Felix-Archemede Pouchet. During this period. Pasteur played a critical and major role in the proper standardization of various processes and techniques intimately associated with the said two ‗alcoholic beverages‘ in order to obtain a consistently good product. Darwin (1859) in his book. who published in 1859 an extensive report ―proving‖ its occurrence. a process known as Tyndallisation. Pasteur vehemently argued and suggested that the unwanted/undesirable types of microbes must be destroyed and removed by heating not enough to alter the original and authentic inherent flavor/aroma of the fruit juice. Pasteur pointed out that no growth took place because dust and germs had been trapped on the walls of the curved necks but if the necks were broken off so that dust fell directly down into the flask. More important there was an acceptance of their work by the scientific community throughout the world and a willingness to continue and expand the work. They carried out a more logical and convincing experimental design by passing air via cotton fibers so as to prevent the bacterial growth . The flask's opening were freely open to the air but curved so that gravity would cause any air borne dust particle to deposit in the lower parts of the neck. we see the real beginning of microbiology as a discipline of biology. etc. Pasteur invented a ―swannecked‖ flask in early 1800‘s. Some of these ingenious little flasks are still on display at the Pasteur Institute in Paris in their original sterile form. which technique being used still as to date. swan neck shaped tubes. 'Origin of the Species' showed that the human body could be conceived as a creature susceptible to the laws of nature. whiskies. No growth occurred even though the contents of the flasks were exposed to the air. Pasteur. The flasks were heated to sterilize the broth and then incubated. champagne. GOLDEN AGE OF MICROBIOLOGY (1857-1914) The Golden age of microbiology began with the work of Louis Pasteur and Robert Koch who had their own research institute and lasted upto the works of Paul Ehrlich. thus in 1861 finally resolved the controversy of spontaneous generation versus biogenesis and proved that microorganisms are not spontaneously generated from inanimate matter but arise from other microorganisms.Experiment of H. dry-wines.
He perfected the technique of isolating bacteria in pure culture. In 1882 he discovered Mycobacterium tuberculosis. (3) Injected bacteria into other animals to observe whether they got also infected. (4) Isolated microbes from experimentally infected animals squarely matched with those obtained originally from sheep that died due to infection of anthrax. He proposed Koch postulate which were published in 1884 and are the corner stone of the germ theory of diseases and are still in use today to prove the etiology (specific cause) of an infectious disease. it was perhaps caused by a germ carried eventually from one mother to another either by midwives or physicians. . Pasteurization is the application of a high heat for a short time to destroy microorganisms. Various scientists supported and proved the aforesaid ‗germ theory‘ in one way or the other as stated under: Girolamo Fracastro (1483–1553): He advocated that certain diseases might be caused by virtue of invisible organisms transmitted from one subject to another. a famous English surgeon is known for his notable contribution to the antiseptic treatment for the prevention and cure of wound infections. he developed a system of antiseptic surgery designed to prevent microorganisms from entering wounds by the application of phenol on surgical dressings and at times it was sprayed over the surgical areas. Robert Koch (1843–1910): Robert Koch gave the first direct demonstration of the role of bacteria in causing disease. Pasteurization was introduced into the United States on a commercial basis in 1892. and subsequently developed clinical symptoms of anthrax. This ‗destructive microbial phenomenon‘ could be accomplished successfully by holding the juices at a temperature of 145°F (≡ 62. Lister concluded that wound infections too were due to microorganisms. Thus Joseph Lister was the first to introduce aseptic techniques for control of microbes by the use of physical and chemical agents which are still in use today. the cause of anthrax) in 1876. Lord Joseph Lister (1827-1912): This man. Plenciz (1762): He stated that the living microbes (or agents) are the ultimate cause of disease but at the same time aired his views that different germs were responsible for different ailments. Oliver Wendell Holmes (1809–1894): He suggested that puerperal fever was highly contagious in nature. besides. Because of these notable contributions. Ignaz Philipp Semmelweis (1818–1865): He pioneered the usage of antiseptics specifically in the obstetrical practices. His work led to the development of the germ theory of disease. (2) Bacteria examined microscopically so as to ascertain only one specific type was present. Germ Theory: ―Microbes (or bacteria) happen to be the root cause of several human dreadful diseases‖ – Germ theory. He also introduced the use of solid culture media in 1881 by using gelatin as a solidifying agent. Joseph Lister is known as the Father of Antiseptic surgery. In 1867. He was a German physician who first of all isolated anthrax bacillus (Bacillus anthracis. Koch adopted the following steps to isolate microbes causing anthrax: (1) First of all these bacteria were duly grown in cultures in the laboratory.percentage of the ‗bad microbial population‘. He also devised a method to destroy microorganisms in the operation theatre by spraying a fine mist of carbolic acid into the air. thus producing an antiseptic environment.8°C) for a duration of 30 minutes (Pasteurization).
Koch smeared bacteria on a sterile glass slide. followed by addition of certain specific dyes so as to observe the individual cells more vividly under a microscope. The suspected microorganisms must be isolated and grown in a pure culture. in 1878. articulated. The container on being subjected to incubation for a definite period gave rise to bacteria of a single type. Example: Lister diluted milk. Pure Culture: Pure culture may be defined as the propagation of microorganisms or of living tissue cells in special media that are conducive to their growth. 5. The same microorganism must be isolated again from the diseased host. comprising of a mixture of bacteria. Classical Laboratory Method and Pure Culture: Laboratory Methods: Well defined.Koch’s postulates 1. and ‗molasses‘. The guinea pigs subsequently died of tuberculosis. such as: gelatin. very much akin to the parent cell. Experiment Koch developed a staining technique to examine human tissue. with a specially designed syringe until a ‗single organism‘ was strategically delivered into a container of sterile milk. Importantly. Colonies: For the specific study of microorganism. & Specific cultures for manufacturing dairy products viz. tuberculosis from the dead guinea pigs and was able to again culture the microbe in pure culture on coagulated blood serum. 3. M. Koch isolated M. 3. 2.. 2. The same disease must result when the isolated microorganism is inoculated into a healthy host. However. tuberculosis into guinea pigs. In other words it may also be explained as the growth of mass of cells belonging to the same species in a laboratory vessel (e. agar into the media in order to obtain characteristic isolated growths of organisms usually called as colonies. a test tube). 4. 4. the development of a liquefiable solid-culture medium proved to be of immense fundamental importance. cheeses.. and explicit laboratory methods have been adequately developed which enable it to isolate a host of microorganisms representing each species. tuberculosis in pure culture on coagulated blood serum. High-yielding alcohol producing strains from ‗malt wart‘. The microorganism must be present in every case of the disease but absent from healthy organisms. each colony is essentially comprised of millions of individual bacterial cells packed tightly together. Microorganisms causing a large number of infections. Lister termed it as Bacterium lactis. who first and foremost could lay hand on pure cultures of bacteria by the aid of serial dilution technique in liquid media. and succeeded in the isolation of Mycobacterium tuberculosis. Certain specific fermentative procedures. from these identified colonies one may transfer pure cultures to other sterile media. 6. Now. Nitrogen-fixation in soil. Koch grew M. . Example: Koch thoroughly examined material obtained from subjects suffering from pulmonary tuberculosis. It was indeed Joseph Lister. Selected good cultures for making top-quality wines. Koch injected cells from the pure culture of M. besides to cultivate each of the species individually. Koch carefully incorporated some specific solidifying agents. Importance of Pure Culture: 1.g. yogurt. tuberculosis cells could be identified in diseased tissue.
inducing them to produce an antitoxin. now known to be antibodies (humoral immunity). white blood cells) were able to ingest (eat up) the disease-producing microorganisms present in the body. pertussis. Elie Metchnikoff: He described for the first time the manner certain specific leukocytes (i.. Jenner in 1798 published his results on 23 successful vaccinators. tetanus. As with Jenner's vaccination for small pox. principle of the preventive treatment of rabies also worked fully which laid the foundation of modern immunization programme against many dreaded diseases like diphtheria. based on the Latin word 'Vacca' meaning cow. Pasteur’s successful rabies vaccine: Jenner's experimental significance was realized by Pasteur who next applied this principle to the prevention of anthrax and it worked. Encouraged by the successful prevention of anthrax by vaccination. Corynebacterium diphtheriae and showed that it produced its toxins (poisons) in a laboratory flask. 1796 he proved that inoculating people with pus from cowpox lesions provided protection against small pox. He was impressed by the observation that countryside milk maid who contacted cowpox (Cowpox is a milder disease caused by a virus closely related to small pox) while milking were subsequently immune to small pox.Immunity: Immunity refers to the state of being immune to or protected from a disease. Edwin Klebs (1883) and Frederick Loeffler (1884): They discovered the diphtheria bacillus. and management of this fatal disease.they cultivated (grown) the microorganism responsible for causing tetanus (lockjaw). On May 14th. a substance in the blood that would inactivate the diphtheria toxin and protect against the disease. and both antitoxins were used in the treatment of people. prevention. The antitoxin work provided evidence that immunity could result from soluble substances in the blood. Von Behring in 1890 reported on immunization against diphtheria by diphtheria antitoxin. The discovery of toxinantitoxin relationship was very important to the development of science of immunology.e. treatment. He . Pasteur marched ahead towards the service of humanity by making a vaccine for hydrophobia or rabies (a disease transmitted to people by bites of dogs and other animals).) De Salmon and Theobald Smith: They proved amply that immunity to a plethora of infectious diseases may be produced quite effectively and efficiently by proper timely inoculation with the killed cultures of the corresponding microorganisms. A tetanus antitoxin was then prepared. Clostridium titani . He called the attenuated cultures vaccines (Vacca = cow) and the process as vaccination.. Interestingly. (Emil von Behring bagged the Nobel Prize in 1901 in physiology or medicine. Shibasaburo Kitasato and Emil von Behring: In 1890. This state is invariably induced by having been exposed to the antigenic marker on an organism that invades the body or by having been immunized with a vaccine capable of stimulating production of specific antibodies.e. antibodies which critically protect against subsequent exposure to the virulent organisms. Eventually this process was known as vaccination. and Behring prepared the corresponding antitoxin for the control. Pasteur‘s practical aspects and Koch‘s theoretical aspects jointly established the fact that the attenuated microorganisms retained their capacity and capability for stimulating the respective host to produce certain highly specific substances i. Thus the use of cow pox virus to protect small pox disease in humans became popular replacing the risky technique of immunizing with actual small pox material. Edward Jenner’s successful cowpox vaccine (1798): An English physician was the first to prevent small pox. Emile Roux (1853-1933) and Alexandre Yersin: The two notable French bacteriologists demonstrated the production of toxin in filtrates of broth cultures of the diphtheria organism. Emil von Behring (1854–1917) and Shibasaburo Kitasato (1852–1931): They injected inactivated toxin into rabbits. especially an infectious disease. polio and measles etc.
Ehrlich's work had laid important foundations for many of the developments to come and the use of Salvarsen marked the beginning of the eni of chemotherapy and the use of chemicals that selectively inhibit or kill pathogens without causing damage to the patient. namely: (a) Antibody & (b) Chemotherapy and Antibiotics Gerhard Domagk: Of Germany in 1935. Domagk was awarded Nobel Prize in 1939 for the discovery of the first sulpha drug. Thus. referred to as cellular immunity. Ehrlich in collaboration with Sakahiro Hata. One day in 1928 upon his return from a week's vacation. Alexander Fleming: Alexander Fleming of England. Fleming noted that the colonies of Staphylococcus bacterium were evidently being destroyed by the nearby Penicillium colonies. Rather than discarding the contaminated plate. a Scottish physician and bacteriologist. molecular biology was born. Subsequently in 1910. he experimented with numerous synthetic dyes and reported that Prontosil. was active against pathogenic. he speculated that the mold was producing a diffusible substance that inhibited the bacterial growth. Jacques and Therese Trefonel: In the same year two French scientists Jacques and Therese Trefonel showed that the compound Prontosil was broken down within the body of the animal to sulfanilamide (Sulfa drug) the true active factor.baptized these highly specific defenders and crusaders against bacterial infections known as phagocytes (‗eating cells‘). and the phenomenon is termed as phagocytosis. The relative simplicity of the microorganism. Thus human blood cells also confer immunity. Streptococci and Staphylococci in mice even though it had no effect against that same infectious agent in a test tube. Paul Ehrlich put forward two altogether newer concepts with regard to the modus operandi whereby the body aptly destroys microorganisms (bacteria). Fleming observed that a plate of Staphylococcus aureus had become contaminated with a green mold Penicillium notatin which had accidentally fallen in plate. Anti = against + bios = life. he found that the dye Trypan Red was active against the trypanosome that causes African sleeping sickness and could be used therapeutically. This dye with antimicrobial activity was referred to as a 'magic bullet'. a red dye used for staining leather. IN 20~ CENTURY: ERA OF MOLECULAR BIOLOGY By the end of 1900. a Japanese physician. introduced the drug Salvarsan (arsenobenzol) as a treatment for syphilis caused by Treponema pallidum. the microbial products that can kill susceptible microorganism and inhibit their growth) penicillin. In the later years the microorganism were picked up as ideal tools to study various life processes and thus an independent discipline of microbiology. Florey and Chain shared the Nobel Prize in 1945 for the discovery and production of penicillin. Observing this plate. The commercial production of penicillin in the USA began in 1941 Fleming. their short life span and the genetic homogeneity provided an authentic simulated model to understand the physiological. Fleming isolated and subcultured the mold for further study. science of microbiology grew up to the adolescence stage and had come to its own as a branch of the more inclusive field of biology. Paul Ehrlich (1854-1915): (Robert Koch‘s brilliant student) In 1904. Metchnikoff’s Theory: Based of the aforesaid explanations Metchnikoff put forward a theory that — ‗the phagocytes were the body‘s first and most important line of defense against a variety of infection‘. biochemical and genetical intricacies of the living organisms. Fleming had been actually interested in searching something that would kill pathogens ever since working on wound infections during the First World War (1914-1918). after the name of the producer organism Penicillium notatium that could destroy several pathogenic bacteria. . Penicillium notatin has been replaced with Penicillium chrysogenum for the commercial production of penicillin. Sir Alexander Fleming in 1929 discovered the first antibiotic (Gr. He extracted from the fungus a compound which he called penicillin.
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