Introduction of microbiology kbk

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Introduction of microbiology kbk

  1. 1. INTRODUCTION OFINTRODUCTION OF MICROBIOLOGYMICROBIOLOGY Laboratory of MicrobiologyLaboratory of Microbiology Faculty of MedicineFaculty of Medicine Brawijaya UniversityBrawijaya University
  2. 2. microbiologymicrobiology The branch of biology that deals withThe branch of biology that deals with microorganisms and their effects on other livingmicroorganisms and their effects on other living organismsorganisms Many branches of Microbiology :Many branches of Microbiology : - Environmental Microbiology- Environmental Microbiology - Space Microbiology- Space Microbiology - Marine Microbiology- Marine Microbiology - Agricultural Microbiology- Agricultural Microbiology - Food Microbiology ……. etc.- Food Microbiology ……. etc. -- Medical MicrobiologyMedical Microbiology
  3. 3. First SessionFirst Session A BRIEF HISTORY OFA BRIEF HISTORY OF MICROBIOLOGYMICROBIOLOGY MICROSCOPEMICROSCOPE
  4. 4. A BRIEF HISTORY OFA BRIEF HISTORY OF MICROBIOLOGYMICROBIOLOGY
  5. 5. THE FIRST OBSERVATION THE DEBATE OVER SPONTANEOUS GENERATION THE GOLDEN AGE OF MICROBIOLOGY THE BIRTH OF MODERN CHEMOTHERAPY : DREAMS OF A “MAGIC BULLET” MODERN DEVELOPMENTS IN MICROBIOLOGY
  6. 6. THE FIRST OBSERVATION  1665 – An Englishman, Robert Hooke, reported to the world that life’s smallest structural units were “little boxes” or “cells” Using his improving version of a compound microscope  he was able to see individual cells  beginning of the cell theory  that all living things are composed of cells  1673 – 1723, Antony van Leeuwenhoek The Dutch merchant and amateur scientist was the first to actually observe live microorganism through his simple single lens microscope  the “animalcules”  in rainwater, in liquid in which peppercorn had soaked, and in material scraped from his teeth He drew the basic form of bacteria as coccus, rods and spiral
  7. 7. coccus rods spiral
  8. 8. THE DEBATE OVER SPONTANEOUS GENERATION  Until the second half of the 19th century many scientists and philosophers (Aristoteles, Samson, Virgil) believed that some form of life could arise spontaneously from nonliving matter  spontaneous generation  People commonly believed that toads, snakes, and mice could be born of moist soil; that flies could emerge from manure; and that maggots could arise from decaying corpses Evidence Pro and Con  1668 – Fransesco Redi, the Italian physician, a strong opponent of spontaneous generation  demonstrate that maggots do not arise spontaneously from decaying meat Redi filled three jars with decaying meat and sealed them tightly. Then he arranged three other jars similarly but left them open. Maggots appeared in the open jars.Sealed containers showed no signs of maggots  Redi’s antagonists were not convinced; they claimed that fresh air was needed for spontaneous generation
  9. 9. THE DEBATE OVER SPONTANEOUS GENERATION  Redi set up a second experiment  three jars were covered with a fine net instead of being sealed. No larvae appeared in the gauze-covered jars  Many scientists still believed that small organisms (van Leeuwenhoek’s animalcules) were simple enough to be generated from nonliving materials  1745 : spontaneous generation seem to be strengthened, when John Needham, an Englishman, found that even after he heated nutrient fluid before pouring them into covered flasks, the cooled solution were soon teeming with microorganisms. Needham claimed that microbes developed spontaneously from the fluid  Twenty years later : Lazzaro Spallanzani, an Italian scientist, suggested that microorganisms from the air probably had entered Needham’s solutions after they were boiled. Spallanzani showed that nutrient fluids heated after being sealed in a flask did not develop microbial growth  Needham responded by claiming the “vital force” necessary for sponta- neous generation had been destroyed by the heat and was kept out of the flasks by the sealed
  10. 10. THE DEBATE OVER SPONTANEOUS GENERATION  Laurant Lavoisier showed the importance of oxygen to life.  Spallanzani’s observations were criticized on the grounds that there was not enough oxygen in the sealed flasks to support microbial life The Theory of Biogenesis  Rudolf Virchow challenged spontaneous generation with the concept of biogenesis, he claim that living cells can arise only from preexisting living cells  Arguments about spontaneous generation continued until 1861, when the issue was resolved by the French scientist Louis Pasteur  With a series of ingenious experiments, Pasteur demonstrate that microorganisms are present in the air and can contaminate sterile solutions, but air itself does not create microbes
  11. 11. Pasteur’s experiments disproving the theory of spontaneous generation (1) Pasteur first poured beef broth into a long-necked flask. (2) Next he heated the neck of the flask and bent it into an S-shaped curve; then he boiled the broth for several minutes. (3) Microorganisms did not appear in the cooled solution, even after long periods
  12. 12.  Pasteur showed that microorganisms can be present in nonliving matter  on solids, in liquids, and in the air  He demonstrated that microbial life can be destroyed by heat  form the basis of aseptic techniques  The debate of Spontaneous Generation disproved  However still have a problem of “spores”  resistant to heat  John Tyndall (1820-1893)  TYNDALLIZATION  Spontaneous Generation Theory totally finished
  13. 13. The Golden Age Of MicrobiologyThe Golden Age Of Microbiology 1857 - 19141857 - 1914 Pasteur & Robert Koch, led to the establishmentPasteur & Robert Koch, led to the establishment of microbiology asof microbiology as a sciencea science Discoveries during these years included both theDiscoveries during these years included both the agents of many diseases and the role ofagents of many diseases and the role of immunity in the prevention and cure of diseaseimmunity in the prevention and cure of disease Fermentation and PasteurizationFermentation and Pasteurization The Germ Theory of DiseaseThe Germ Theory of Disease VaccinationVaccination
  14. 14. Fermentation & PasteurizationFermentation & Pasteurization A group of French merchants asked Pasteur to find out whyA group of French merchants asked Pasteur to find out why wine and beer souredwine and beer soured  a method that would preventa method that would prevent spoilage ?spoilage ? Many scientist believed that air converted the sugars inMany scientist believed that air converted the sugars in these fluids into alcoholthese fluids into alcohol Pasteur found instead that microorganisms calledPasteur found instead that microorganisms called yeastsyeasts convert the sugars to alcohol in the absence of air. Thisconvert the sugars to alcohol in the absence of air. This process, calledprocess, called fermentationfermentation, is used to make wine and, is used to make wine and beerbeer Souring and spoilage are caused by different micro-Souring and spoilage are caused by different micro- organisms calledorganisms called bacteriabacteria. In the presence of air, bacteria. In the presence of air, bacteria change the alcohol in the beverage into vinegar (acetic acid)change the alcohol in the beverage into vinegar (acetic acid)
  15. 15. The Germ Theory of DiseaseThe Germ Theory of Disease Before the time of Pasteur, effective treatments for manyBefore the time of Pasteur, effective treatments for many diseases were discovered by trial and error, but the causesdiseases were discovered by trial and error, but the causes of the disease were unknownof the disease were unknown The realization that yeasts play a crucial role inThe realization that yeasts play a crucial role in fermentation was the first link between the activity of afermentation was the first link between the activity of a microorganism and physical and chemical changes inmicroorganism and physical and chemical changes in organic materialsorganic materials This discovery alerted scientists to the possibility thatThis discovery alerted scientists to the possibility that microorganisms might have similar relationships with plantsmicroorganisms might have similar relationships with plants animals - specifically, that microorganisms might causeanimals - specifically, that microorganisms might cause disease. This idea was known asdisease. This idea was known as the germ theory ofthe germ theory of diseasedisease
  16. 16. The Germ Theory of DiseaseThe Germ Theory of Disease 1840 –1840 – Ignaz SemmelweisIgnaz Semmelweis a Hungarian physician, hada Hungarian physician, had demonstrated that physician, who at the time did notdemonstrated that physician, who at the time did not disinfect their hands, routinely transmitted infectionsdisinfect their hands, routinely transmitted infections (puerperal, or childbirth fever) from one obstetrical patient to(puerperal, or childbirth fever) from one obstetrical patient to anotheranother 1860 –1860 – Joseph ListerJoseph Lister an English surgeon, applied the german English surgeon, applied the germ theory to medical procedure. Disinfectants were not used attheory to medical procedure. Disinfectants were not used at the time, but Lister knew that phenol (carbolic acid) killsthe time, but Lister knew that phenol (carbolic acid) kills bacteria, so he began treating surgical wounds with abacteria, so he began treating surgical wounds with a phenol solutionphenol solution 1876 –1876 – Robert KochRobert Koch, a German physician, proved that, a German physician, proved that bacteria actually cause diseasebacteria actually cause disease  discovered rod-shapeddiscovered rod-shaped bacteria now known asbacteria now known as Bacillus anthracisBacillus anthracis in the blood ofin the blood of cattle that had died of anthraxcattle that had died of anthrax
  17. 17. The Germ Theory of DiseaseThe Germ Theory of Disease Koch’s research provides a framework for the study of theKoch’s research provides a framework for the study of the etiology of any infectious diseaseetiology of any infectious disease  Koch Postulates :Koch Postulates : 1. The same pathogen must be present in every case of1. The same pathogen must be present in every case of diseasedisease 2. The pathogen must be isolated from the diseased host2. The pathogen must be isolated from the diseased host and grown in pure cultureand grown in pure culture 3. The pathogen from the pure culture must cause the3. The pathogen from the pure culture must cause the disease when it is inoculated into a healthy, susceptibledisease when it is inoculated into a healthy, susceptible laboratory animallaboratory animal 4. The pathogen must be isolated from the inoculated4. The pathogen must be isolated from the inoculated animal and must be shown to be the original organismanimal and must be shown to be the original organism
  18. 18. VACCINATIONVACCINATION 1796 –1796 – Edward JennerEdward Jenner, a young British physician, embarked, a young British physician, embarked on an experiment to find a way to protect people fromon an experiment to find a way to protect people from smallpox :smallpox : - A young milkmaid informed Jenner that she couldn’t get- A young milkmaid informed Jenner that she couldn’t get smallpox because she already had been sick from cowpoxsmallpox because she already had been sick from cowpox - First Jenner collected scrapings from cowpox blisters. Then- First Jenner collected scrapings from cowpox blisters. Then he inoculated a healthy 8-year-old volunteer with thehe inoculated a healthy 8-year-old volunteer with the cowpox material by scratching the person’s arm with acowpox material by scratching the person’s arm with a pox-contaminated needle. In a few days, the volunteerpox-contaminated needle. In a few days, the volunteer became mildly sick but recovered and never againbecame mildly sick but recovered and never again contracted either cowpox or smallpox. The process wascontracted either cowpox or smallpox. The process was calledcalled vaccinationvaccination (vacca=cow)(vacca=cow) - The protection from disease provided by vaccination is- The protection from disease provided by vaccination is calledcalled immunityimmunity
  19. 19. VACCINATIONVACCINATION 1880 – Pasteur discovered why vaccination work. He found1880 – Pasteur discovered why vaccination work. He found that the bacteria that causesthat the bacteria that causes fowl cholerafowl cholera lost its abilitylost its ability to cause disease after it was grown in the laboratory forto cause disease after it was grown in the laboratory for long periods. However it – and other microorganisms withlong periods. However it – and other microorganisms with decreased virulence – was able to induce immunity againstdecreased virulence – was able to induce immunity against subsequent infections by its virulent counterpart.subsequent infections by its virulent counterpart. Some vaccines are still produced from avirulent microbialSome vaccines are still produced from avirulent microbial strains that stimulate immunity to the related virulent strain.strains that stimulate immunity to the related virulent strain. Other vaccines are made from killed virulence microbes,Other vaccines are made from killed virulence microbes, from isolated components of virulent micoorganisms, or byfrom isolated components of virulent micoorganisms, or by genetic engineering techniquesgenetic engineering techniques
  20. 20. The Birth of Modern Chemotherapy :The Birth of Modern Chemotherapy : Dreams of a “ Magic Bullet “Dreams of a “ Magic Bullet “ After the relationship between microorganisms andAfter the relationship between microorganisms and disease was established, medical microbiologists nextdisease was established, medical microbiologists next focused on the search for substances that could destroyfocused on the search for substances that could destroy pathogenic microorganisms without damaging thepathogenic microorganisms without damaging the infected animal or humaninfected animal or human Treatment of disease by using chemical substances isTreatment of disease by using chemical substances is calledcalled chemotherapychemotherapy Chemotherapeutic agents prepared from chemicals inChemotherapeutic agents prepared from chemicals in the laboratory are calledthe laboratory are called synthetic drugssynthetic drugs Chemicals produced naturally by bacteria or fungi to actChemicals produced naturally by bacteria or fungi to act against other microorganisms are calledagainst other microorganisms are called antibioticsantibiotics
  21. 21. The First Synthetic DrugsThe First Synthetic Drugs Paul EhrlichPaul Ehrlich, a German physician, was the imaginative, a German physician, was the imaginative thinker who fired the first shot in the chemotherapythinker who fired the first shot in the chemotherapy revolution. Ehrlich speculated about a “magic bullet” thatrevolution. Ehrlich speculated about a “magic bullet” that could destroy pathogen without harming the infected hostcould destroy pathogen without harming the infected host In 1910, he found a chemotherapeutic agent calledIn 1910, he found a chemotherapeutic agent called salvarsansalvarsan, an arsenic derivative effective against syphylis, an arsenic derivative effective against syphylis By the late 1930s, researchers had developed several otherBy the late 1930s, researchers had developed several other synthetic drugs, mostly were derivatives of dyes that couldsynthetic drugs, mostly were derivatives of dyes that could destroy microorganismsdestroy microorganisms In addition,In addition, Domagk (1935)Domagk (1935) discovered thatdiscovered that prontosilprontosil hadhad dramatically effect against streptococcal infectionsdramatically effect against streptococcal infections  in thein the body was changed intobody was changed into sulfanilamidesulfanilamide that analog withthat analog with PABAPABA
  22. 22. A Fortunate Accident - AntibioticsA Fortunate Accident - Antibiotics The first antibiotic was discovered by accidentThe first antibiotic was discovered by accident 1928 –1928 – Alexander FlemingAlexander Fleming, a Scottish physician and, a Scottish physician and bacteriologist, looked at the curious pattern of growth on thebacteriologist, looked at the curious pattern of growth on the contaminated plates. There was a clear area around thecontaminated plates. There was a clear area around the mold where the bacterial culture had stopped growingmold where the bacterial culture had stopped growing The mold was later identified asThe mold was later identified as Penicillium notatumPenicillium notatum, and, and Fleming named the mold’s active inhibitor asFleming named the mold’s active inhibitor as penicillinpenicillin.. The enormous usefulness of penicillin was not apparentThe enormous usefulness of penicillin was not apparent until the 1940suntil the 1940s  Florey & ChainFlorey & Chain 1939 –1939 – ReneRene´´ Dubos,Dubos, a French microbiologist, discovereda French microbiologist, discovered two antibiotics calledtwo antibiotics called gramicidingramicidin andand tyrocidine.tyrocidine. Both wereBoth were produced by a bacterium,produced by a bacterium, Bacillus brevisBacillus brevis, cultured from soil., cultured from soil.
  23. 23. Modern Development in MicrobiologyModern Development in Microbiology BacteriologyBacteriology MycologyMycology ParasitologyParasitology ImmunologyImmunology VirologyVirology Recombinant DNA technologyRecombinant DNA technology
  24. 24. VIROLOGYVIROLOGY The study of virus, actually originated during the Golden Age ofThe study of virus, actually originated during the Golden Age of MicrobiologyMicrobiology 1892 –1892 – Dmitri IwanowskiDmitri Iwanowski reported that the organism that causereported that the organism that cause mosaic disease of tobacco was so small that it passed through filtermosaic disease of tobacco was so small that it passed through filter fine enough to stop all known bacteriafine enough to stop all known bacteria 1935 –1935 – Wendell StanleyWendell Stanley demonstrated that the organism, calleddemonstrated that the organism, called tobacco mosaic virus (TMV)tobacco mosaic virus (TMV), was fundamentally different from other, was fundamentally different from other microbes and so simple and homogeneous that it could be cristallizedmicrobes and so simple and homogeneous that it could be cristallized like chemical compoundlike chemical compound 1940 – Since the development of the electron microscope,1940 – Since the development of the electron microscope, microbiologists have been able to observe the structure of viruses inmicrobiologists have been able to observe the structure of viruses in detail.detail.
  25. 25. Recombinant DNA TechnologyRecombinant DNA Technology Until the 1930s, all genetic research was based on the study of plant andUntil the 1930s, all genetic research was based on the study of plant and animal cellsanimal cells In the 1940s, the scientists turned to unicellular organisms, primarilyIn the 1940s, the scientists turned to unicellular organisms, primarily bacteriabacteria In 1941 –In 1941 – George W.BeadleGeorge W.Beadle andand Edward L.TatumEdward L.Tatum demonstrated thedemonstrated the relationship between genes and enzymesrelationship between genes and enzymes 1944 –1944 – Oswald Avery, Colin McLeodOswald Avery, Colin McLeod, and, and Maclyn McCartyMaclyn McCarty  DNA wasDNA was established as the hereditary materialestablished as the hereditary material 1946 –1946 – Joshua LederbergJoshua Lederberg andand Edward L.TatumEdward L.Tatum discovered that geneticdiscovered that genetic material could be transferred from one bacterium to another by a processmaterial could be transferred from one bacterium to another by a process called conjugationcalled conjugation 1953 –1953 – James WatsonJames Watson andand Francis CrickFrancis Crick proposed a model for theproposed a model for the structure and replication of DNAstructure and replication of DNA
  26. 26. MICROSCOPEMICROSCOPE
  27. 27. MICROSCOPEMICROSCOPE Compound Light MicroscopyCompound Light Microscopy Darkfield MicroscopyDarkfield Microscopy Phase-contrast MicroscopyPhase-contrast Microscopy Fluorescence MicroscopyFluorescence Microscopy Confocal MicroscopyConfocal Microscopy Electron MicroscopyElectron Microscopy
  28. 28. Compound Light MicroscopeCompound Light Microscope Has a series of lenses and use visible light as its sourceHas a series of lenses and use visible light as its source of illuminationof illumination A series of finely ground lenses forms a clearly focusedA series of finely ground lenses forms a clearly focused image that is many time larger than the specimens itselfimage that is many time larger than the specimens itself This magnification is achieved when light rays from anThis magnification is achieved when light rays from an illuminatorilluminator  condensorcondensor  specimenspecimen  objectiveobjective lenseslenses  ocular lensocular lens Total magnification = objective lens magnification xTotal magnification = objective lens magnification x ocular lens magnificationocular lens magnification Objective lens : 10 x (low power), 40 x (high power), andObjective lens : 10 x (low power), 40 x (high power), and 100 x (oil immersion)100 x (oil immersion) Ocular lens : 10 xOcular lens : 10 x
  29. 29. Compound Light MicroscopeCompound Light Microscope The total magnifications would be 100 x for low power,The total magnifications would be 100 x for low power, 400 x for high power , and 1000 x for oil immersion.400 x for high power , and 1000 x for oil immersion. Some compound light microscopes can achieve aSome compound light microscopes can achieve a magnification of 2000 x with the oil immersion lens.magnification of 2000 x with the oil immersion lens. The oil immersion has the same refractive index asThe oil immersion has the same refractive index as glass, so the oil becomes part of the optics of the glassglass, so the oil becomes part of the optics of the glass of the microscope.of the microscope. Unless immersion oil is used, light rays are refracted asUnless immersion oil is used, light rays are refracted as they enter the air from the slidethey enter the air from the slide
  30. 30. Compound Light MicroscopeCompound Light Microscope
  31. 31. Darkfield MicroscopyDarkfield Microscopy Is used for examining live microorganisms that either areIs used for examining live microorganisms that either are invisible in the ordinary light microscope, cannot beinvisible in the ordinary light microscope, cannot be stained by standard methods, or are so distorted bystained by standard methods, or are so distorted by staining that their characteristics then cannot be identifiedstaining that their characteristics then cannot be identified A darkfield microscope uses a darkfield condensor thatA darkfield microscope uses a darkfield condensor that contain an opaque disccontain an opaque disc  the specimen appears lightthe specimen appears light against a black backgroundagainst a black background One use for darkfield microscopy is the examination ofOne use for darkfield microscopy is the examination of very thin spirochetes, such asvery thin spirochetes, such as Treponema pallidumTreponema pallidum, the, the causative agent of syphiliscausative agent of syphilis
  32. 32. Phase-Contrast MicroscopyPhase-Contrast Microscopy Is especially useful because it permits detailed examinationIs especially useful because it permits detailed examination of internal structures in living microorganismsof internal structures in living microorganisms The principle of phase-contrast microscopy is based on theThe principle of phase-contrast microscopy is based on the wave nature of light rays, and the fact that light rays can bewave nature of light rays, and the fact that light rays can be in phasein phase (their peaks and valleys match) or(their peaks and valleys match) or out of phaseout of phase.. In phase-contrast microscopy, one set of light rays comesIn phase-contrast microscopy, one set of light rays comes directly from the light source. The other set comes from lightdirectly from the light source. The other set comes from light that is reflected or diffracted from particular structure in thethat is reflected or diffracted from particular structure in the specimenspecimen Phase-contrast microscope is provided with diffraction platePhase-contrast microscope is provided with diffraction plate
  33. 33. Brightfield Darkfield Phase-contrastBrightfield Darkfield Phase-contrast
  34. 34. Fluorescence MicroscopyFluorescence Microscopy The object is stained with one of a group of fluorescent dyesThe object is stained with one of a group of fluorescent dyes calledcalled fluorochromesfluorochromes ((primolin, acridine orange R, thiazo yellow-G, auramine O, fluorescein isothiocyanate) The light source  Ultraviolet light The principal use of fluorescence microscopy is a diagnostic technique called fluorescence-antibody (FA) technique, or immunofluorescence
  35. 35. Confocal MicroscopyConfocal Microscopy A fairly recent development in light microscopyA fairly recent development in light microscopy Like fuorescent microscopy, specimens are stained withLike fuorescent microscopy, specimens are stained with fluorochromesfluorochromes The light sourceThe light source  laserlaser Most confocal microscopes are used in conjunction withMost confocal microscopes are used in conjunction with computers to construct three-dimensional imagescomputers to construct three-dimensional images Can be used to evaluate cellular physiology byCan be used to evaluate cellular physiology by monitoring the distributions and concentrations ofmonitoring the distributions and concentrations of substances such as ATP and calcium ions.substances such as ATP and calcium ions.
  36. 36. Electron MicroscopyElectron Microscopy Object smaller than about 0.2Object smaller than about 0.2 µm, such as virus, or theµm, such as virus, or the internal structures of cels must be examined with aninternal structures of cels must be examined with an electron microscopeelectron microscope A beam of electrons is used instead of lightA beam of electrons is used instead of light Objects are generally magnified 10,000 – 100,000 xObjects are generally magnified 10,000 – 100,000 x Instead of using glass lenses, an electron microscopeInstead of using glass lenses, an electron microscope uses electromagnetic lenses to focus a beam ofuses electromagnetic lenses to focus a beam of electrons onto specimenelectrons onto specimen There are two types of electron microscope :There are two types of electron microscope : - The transmission electron microscope- The transmission electron microscope - The scanning electron microscope- The scanning electron microscope

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