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The Science of Microbiology. Teaching Objectives. To define Microbiology To be familiar with history of Microbiology To introduce the following: The extent of the microbial world Microbial phylogenetic and metabolic diversity Interaction of microbes and hosts
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The Science of Microbiology . H.Gh.Safaei
Teaching Objectives • To define Microbiology • To be familiar with history of Microbiology • To introduce the following: • The extent of the microbial world • Microbial phylogenetic and metabolic diversity • Interaction of microbes and hosts • Prokaryotic characteristic • Application of microbiology
History of Microbiology • Antoni van Leeuwenhoek, DE (1673) • First observed live microorganisms (animalcules) • Schleiden and Schwann, DE • Formulated Cell Theory: cells are the fundamental units of life and carry out all the basic functions of living things • Pasteur, FR and Tyndall, UK (1861) • Finally disproved S.G.
DEFINITIONS of Microbiology Microbiology is the study of microorganisms H.Gh.Safaei
5 x 1030 microbial cells exist on earth; cells constitute about 90% of the biomass of the entire biosphere. more than 90% of the cells in our bodies are microbes H.Gh.Safaei
biologic diversity • Biochemistry, molecular biology, and genetics • The number of genes within a prokaryote may vary 468 in Mycoplasma genitalium 7825 in Streptomyces coelicolor, many of these genes must be dedicated to essential functions such as energy generation, macromolecular synthesis, and cellular replication H.Gh.Safaei
Microorganisms of each cell type : 1. Eukaryotic - fungi, protozoa, algae, worms 2. Prokaryotic - bacteria 3. Viruses - neither – a cellular 4. number of transmissible plant diseases are caused by viroids 5.Prion, scrapie, a degenerative central nervous system disease of sheep H.Gh.Safaei
ECOLOGICAL RELATIONSHIPS • SYMBIOSIS: neutral, antagonistic or synergistic relationship between two dissimilar organisms (SYMBIOTES, SYMBIONTS) living in close association with each other; • MUTUALISM (+/+): mutually beneficial relationship between two species • COMMENSALISM (+/0): relationship between two species in which one is benefited and the other is not affected, neither negatively nor positively • PARASITISM (+/-): relationship between two species in which one benefits (parasite) from the other (host); usually involves detriment to the host ,e.g. Mycoplasmas, chlamydiae and rickettsiae H.Gh.Safaei
DIVISIONS OF MICROBIOLOGY • Organisms studied • 1. Bacteriology - study of bacteria • 2. Mycology - study of fungi • 3. Phycology - study of algae • 4. Protozoology - study of protozoa • 5. Virology - study of viruses • 6. Parasitology - study of parasites H.Gh.Safaei
Prokaryote • small size, usually on the order of 1 micron in diameter, and the absence of a nuclear membrane • nucleoid and can be visualized by electron microscopy • The DNA of almost all bacteria is a circle with a length of about 1 mm H.Gh.Safaei
Prokaryotic Communities • A useful survival strategy for specialists is to enter into consortia • Arrangements contribute to survival of the group as a whole • clone that may contain up to 108 cells • Genetic variability • Extracellular polysaccharides H.Gh.Safaei
Health related 1. Etiology - identification of causative agent of disease 2. Epidemiology - study of spread , of disease 3. Immunology - study of immune system 4. Chemotherapy - treatment of disease with chemical compounds 5. Infection control - control of spread of infectious disease H.Gh.Safaei
Processes, Functions 1. Microbial metabolism 2. Microbial genetics 3. Microbial ecology H.Gh.Safaei
BENEFICIAL APPLICATIONS (WHY STUDY MICROBIOLOGY?) A. Causative agents of infectious diseases B. Normal flora 1. Beneficial metabolic functions 2. Antagonistic effect - prevents invasion pathogens, over growth of potential pathogens3. Normal flora vs. identification pathogen C. Environmental importance 1. Decomposers 2. Produce oxygen 3. Food chain 4. Sewage treatment H.Gh.Safaei
Industrial importance • 1. Food industry • 2. Brewing industry • 3. Pharmaceutical industry • 4. Genetic engineering H.Gh.Safaei
Research - genetics, metabolism • Simple cell structure • Rapid rate of growth • Inexpensive to culture H.Gh.Safaei
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BACTERIAL Taxonomy , Classification Cell Structure , Genetics, Metabolism H.Gh.Safaei
Objective • To define the Taxonomy , Classification , • To be familiar with Classification of micro organism • To explain the cell structure of bacteria ( cell wall , cytoplasmic membrane, spore, flagella, capsule , gentics and metabolism ) H.Gh.Safaei
DEFINITIONS • A. Taxonomy - study of classification. • B. Classification - arranging organisms into related groups H.Gh.Safaei
Criteria used for bacterial classification & identification : 1. Morphology • a. Colony • b. Microscopic (cell) 2. Staining reactions (Gram stain - cell wall chemistry) 3. Physiological characteristics • a. Growth requirements - temperature, O2, pH, etc. • b. Nutritional requirements - biochemical activity (enzymes) • c. Structure of DNA • d. Antigenic structure - serological tests H.Gh.Safaei
Bacterial cell structure • Cell wall • Cell membrane • Cytoplasm (Ribosome, Vacuoles, Mesosome ) • Nucleoid • Flagella • Pilli • Capsule • Spore H.Gh.Safaei
Microorganisms of each cell type 1. Eukaryotic - fungi, protozoa, algae, Slime mold 2. Prokaryotic - bacteria 3. Viruses - neither – acellular H.Gh.Safaei
Archaea (archaebacteria), • The Bacteria (eubacteria), H.Gh.Safaei
PROKARYOTIC CELLS 1. No "true" nucleus - nucleoid. • a. No nuclear membrane. • b. No paired chromosomes. • c. No histones. • d. No nucleolus. 2. No organelles. 3. No cytoplasmic streaming. 4. Cell membrane contains no sterols. 5. Cell walls • a. All typical prokaryotic cells possess cell walls. • b. Composed of peptidoglycan (murein). 6. Division - binary fission. H.Gh.Safaei
System of Classification 1. Based on following properties: • a. Type of cell - eukaryotic, prokaryotic • b. Structure - multicellular or unicellular • c. Mode of nutrition / metabolism - ingestion, absorption, photosynthesis • d. Structure DNA ( Sequence , % G+C) H.Gh.Safaei
Characteristics of the organism : • a. Size - Micrococcus • b. Shape - Staphylococcus, Bacillus • c. Arrangement - Staphylococcus, Streptococcus • d. Color of colony - Micrococcus roseus • e. Habitat - Escherichia coli, Staphylococcus epidermidis • f. Nutritional requirements - Haemoplilusaegypticus • g. Disease - Neisseriameningitidis, N. gonorrhoeae, C. diphtheriae, V. cholerae • h. Researcher - Escherichia, Neisseria H.Gh.Safaei
Cell Wall H.Gh.Safaei
Cell division H.Gh.Safaei
Binary vision H.Gh.Safaei
Binary Division • 1 to 2 to 4 to 8 to ? H.Gh.Safaei
Bacterial growth • Bacterial growth is increase in cell numbers • Growth depends on temperature, pH, osmotic pressure, oxygen, and nutrients • Log phase cultures are most sensitive to growth inhibition H.Gh.Safaei
The growth curve • Lag phase • exponential phase: balanced growth • Stationery phase • Death • Bacterial cell cycle and cell division H.Gh.Safaei
Growth measurements: direct • Plate counts • detect viable organisms • Direct microscopic counts • detect total numbers H.Gh.Safaei
Measuring Growth Turbidity Spectrophotometer Scale %Transmittance Optical Density or Absorbance Filtration 0.45 - 0.2 um sizes Grid Pattern on Filter Standards for Public Health 0 E.coli / 100 ml of water Also used for sterilization H.Gh.Safaei
Requirements for bacterial growth • Definition of growth • New “state”: VBNC- viable but not culturable • Nutritional requirements: water, carbon, nitrogen, phosphorous, sulfur H.Gh.Safaei
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Bacterial growth • Physical requirements • pH • Temperature: Psychrophiles, Mesophiles- pathogens, Thermophiles • Osmotic pressure • Oxygen • Nutrients H.Gh.Safaei
Temperature Problems • Thermophiles • Hot tubs-public • Heat Resistant Enzymes in PCR • Psychrophiles • Cold food (growth in transit) • Listeria grows in hot dogs H.Gh.Safaei
Factors that Influence Growth • pH • Acidophiles: • Grow optimally between ~pH 0 and 5.5 • Neutrophiles • Growoptimally between pH 5.5 and 8 • Alkalophiles • Grow optimally between pH 8 – 11.5 H.Gh.Safaei
Physical Factors: • Moisture • Oxygen • Aerobes and Anaerobes • Osmotic Pressure • Pseudomonas grows in distilled water • Jelly- Jams (high sugar) • Salt (high salt) H.Gh.Safaei
Nutritional Factors • Carbon Source • Auto- or Hetero- • CO2 or Organic compounds • Nitrogen Source • NO3- ==> NO2 ==> NH3 to Amino Acids ==> Proteins • Sulfur • Phosphorus • Trace Elements • Fe, some required • Host Sideophores bind Fe H.Gh.Safaei