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Edit Comment Close Premium member Presentation Transcript Medical Microbiology : Medical Microbiology 醫學微生物這門課包含的內容繁多, 千頭萬緒. 在 Murray PR et al. 編寫的 Medical Microbiology (第四版) 所寫的序言裡說: 學生最好能 " Thinking like a physician", 要常常問七個問題: Who, Where, When, Why, Which, What and How. 醫學微生物的內容不只是應用醫學的一項, 它也是一門自然科學. 它也有科學思辨的趣味, 知識不只是工具而已. 學生還可以 “ Thinking like a scientist”. 要常常問古早人是怎麼想的? 問題的困難在哪裡? 有沒有比目前的解釋更好的答案? 基本知識架構可依自己所長加以斟酌. 上課內容在網頁上:http://myweb.ncku.edu.tw/~y1357/course/micro.htmlScale of life: Scale of life BC Yang Paramecium HIV virion Animal Cell Bacterial Cell In mm3 Prion 27-30 kDa 20-50,000 0.01-50 0.00001-0.01 Pox group Photosynthetic bacteria Unicellular algae Viroid ~400bp For lecture onlyDiscovery of microbes: Discovery of microbes BC Yang Antony van Leeuwenhoek My work, which I've done for a long time, was not pursued in order to gain the praise I now enjoy, but chiefly from a craving after knowledge, which I notice resides in me more than in most other men. And therewithal, whenever I found out anything remarkable, I have thought it my duty to put down my discovery on paper, so that all ingenious people might be informed thereof. Antony van Leeuwenhoek. (1632-1723) Letter of June 12, 1716 Robert Hooke 1635-?, (cell theory) For lecture onlyA short history of microbiology: A short history of microbiology On spontaneous generation: 1668, Francesco Redi (with cover, against); 1749, John Needham (simple boiling, for); 1765, Lazzaro Spallanzani (extensive boiling, against) 1673: Anton van Leeuwenhoek (microbes) 1678: Robert Hooke (cell structure) 1861: Louies Pasteur (gooseneck bottle; against) 1870: Joseph Lister (1% carbolic acid, phenol in surgical procedure) 1881: Robert Koch (Koch’s postulate) 1892: Dmitrii Iwanowski (Tobacco Mosaic Virus) 1909: Paul Ehrlich (chemotherapy, arsenicals against spirochetes) 1915: Frederick W. Twort (bacteriophage) 1940: O.T. Avery (DNA as transforming factor) 1953: James D. Watson & Francis H.C. Crick (DNA structure) BC Yang For lecture onlyBig person in microbiology: Big person in microbiology Louis Pasteur, 1822-1895, French BC Yang For lecture onlyL. Pasteur at "scientific evening" of the Sorbonne University in Paris: L. Pasteur at "scientific evening" of the Sorbonne University in Paris " And I wait, I watch, I question it!-begging it to recommence for me the beautiful spectacle of the first creation. But it is dumb, dumb since these experiments were begun several years ago; it is dumb because I have kept it from the only thing the human mind does not know how to produce: from the germs which float in the air, from Life, for life is a germ and a germ is life.“ " The germ theory is not a philosophical theory of life, but merely a body of factual observations from a series of practical operations." Pasteur's own drawings of the swan-necked flasks BC Yang For lecture onlyBig person in microbiology: Big person in microbiology Rober Koch,1843-1910, Germany BC Yang For lecture onlyCommunication and classification: Communication and classification By making photomicrobiographs, I can reveal the bacteria true to nature and free of subjective misinterpretation ---Rober Koch BC Yang For lecture onlySlide9: Privy Health Officer Berlin W, 15, June 30, 1904 Dear Sir, I still remember very well the first time I took an oil system into my hands and myself witnessed the dramatic progress achieved by the optical workshop of Carl Zeiss under the direction of Professor Abbe's ingenious advisory board. Every time I subsequently used oil systems, I thought with admiration and gratitude of the magnificent gift made by the Zeiss optical workshop to everyone working with a microscope. After all, I owe a large percentage of the success I have had the privilege of achieving for science to your excellent microscopes which were of enormous benefit to me in the expedition to South Africa which I have just completed. Respectfully yours, R. Koch http://www.zeiss.com/C12567A100537AB9/allBySubject/DAF3D66784516F2CC1256B45003DA810 For lecture only , BC YangAn obvious true is never easy to be recognized and accepted: An obvious true is never easy to be recognized and accepted Koch’s demonstrations in the Institute of plant pathology; University of Breslau (15 Nov. 1876) Now leave everything as it is, and go to Koch, This man has made a magnificent discovery ----Julius Cohnheim 1817-1891 By making photomicrographs, I can reveal the bacteria true to nature and free of subjective misinterpretation ---- Robert Koch 1843-1910 Whole business seemed quite improbable; Anything I couldn’t see with a dry lens wasn’t worth looking at. ----Rudolf Virchow 1821-1902 BC Yang For lecture onlyTwo key factors: Two key factors Improvement of optical instruments Establishment of pure culture system For lecture only BC YangMicrobial classification and structure (1): Microbial classification and structure (1) BC Yang Yersinia pestis Description of bacteria (G+/G-) Bacillus sp. For lecture only叫不出名字就無法溝通: 叫不出名字就無法溝通 Phenotypic classification Analytic classification (composition) Genotypic classification (gene constitute) For lecture only BC YangMorphological study of bacteria: Morphological study of bacteria Culture techniques Instruments: Light microscope: 0.2 mm Phase-contrast; dark field Electron microscope: 0.0001 mm TEM; SEM Near-field microscope: atom-level Determining factors: Species Growth condition/Culture technique Staining: simple or differential BC Yang For lecture onlyGrowth/Culture condition: Growth/Culture condition Plating technique of Robert Koch for single colony isolation The majority of microbes persist attached to surfaces within a structured biofilm ecosystem and not as free floating organisms. Microbiol. Mol. Biol. Rev. 64:847-867 (2000) BC Yang For lecture onlyDifferences in culture: when cells grow as biofilm: Differences in culture: when cells grow as biofilm 1. On metabolism 2. Drug resistance BC Yang For lecture onlyTwo types of biofilms: Two types of biofilms Symbioses Termite, ruminant digestion Sewage treatment bioreactors Water pipes Dental units Contact lens cases Dental plaque Endocarditis Cystic Fibrosis Otitis media Urinary catheter Implants Disease-associated Environmental BC Yang For lecture onlyClassification: Classification Phenotypic classification Morphology; Biotyping; Serotyping; phage typing; Antibiogram…. Analytic classification Cell wall fatty acid; lipid; protein; enzyme isomers….. Genotypic classification G/C ratio; DNA sequence compare; DNA/RNA hybridization, plasmid…. BC Yang For lecture onlyMorphological study: Morphological study Unstained General shape, motility, spore, intracellular granules Fix and stained Simple stain: Positive: methylene blue, methyl violet, basic fuchsin, silver… Negative: acidic dye for background… Gram’s stain: (by Hans C. Gram, 1884) base on the resistance to decoloration with acetone, alcohol or aniline oil after stained with a triphenyl methane dye. (+: purple; -:red) Acidfast stain reaction: (Ziehl-Neelsen method) cell wall consists of lipids and forms a special anatomical disposition of the lipids. Immunostaining: labeled antibodies BC Yang For lecture onlySlide20: Gram stain (cell wall structure) Mole percent G+C in the genome Growth temperature Ability to form heat stable spores Electron acceptors for respiration Photosynthetic ability Motility Cell shape Ability to use various carbon and nitrogen sources Special nutritional requirements The prokaryotes and eukaryotic protists were more difficult to classify. They are much simpler, and there are few obvious features. Classifications based upon features such as these were not natural systems (Bergey's Manual of Determinative Bacteriology ) BC Yang For lecture onlyGeneral description of bacteria: General description of bacteria Found in most habitats Most numerous organisms on earth Earliest life forms (fossils date 2.5 billion years old) Most are unicellular Most bacteria grow best at a pH of 6.5 to 7.0 Main decomposers of dead organisms so recycle nutrients Some bacteria breakdown chemical & oil spills Some cause disease Classified by their structure, motility (ability to move), molecular composition, & reaction to stains (Gram stain) Grouped into 2 kingdoms --- Eubacteria (true bacteria) & Archaebacteria (ancient bacteria); that once grouped together in the kingdom Monera BC Yang For lecture onlyClassification: Tree of evolution: Classification: Tree of evolution BC Yang For lecture onlyClassification: can you remember all the names?: Classification: can you remember all the names? 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