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Chapter 6: Microbial Growth * Refer to pictures The Requirements for Growth y Physical requirements o Temperature    o o y pH osmotic pressure Minimum growth temperature Optimum growth temperature Maximum growth temperatue

Chemical Requirements o C,N, S, P, trace elements, oxygen, organic growth factor


Plasmolysis (Fig. 6.4)

Psychropiles/psychotrophes y y pH y y y Most bacteria grow bw pH 6.5-7.5 Molds and yeasts grow bw pH 5-6 Acidophiles grow in acidic environment Grow between 0-15C and 10-25C Cause food spoilage

Osmotic Pressure -Hypertonic environments or an increase in slat or sugar cause plasmolysis -Extreme or obligate halophiles require high osmotic pressure (ie 30% salt ± Great Sea, Red Sea) -Facultative halophiles can tolerate high osmotic pressure (2%-15% salt) What would happen if cell put into distilled water? Plasmolysis*

Chemical Requirements for Bacterial Growth Carbon y y y Nitrogen y y y Amino acids and proteins Most bacteria decompose proteins (recycle) Some bacteria use ammonium ions NH4 or nitrate ions Structural organic molecules, energy source Chemoheterotrophs use organic carbon sources Autotrophs use CO2

y Sulfur y y y Phosporus y y

A few bacteria(cyanobacteria) use N2 in nitrogen fixation

In amino acids, thiamine, and biotin Most bacteria decompose proteins to get sulfur Some bacteria use SO4-2 or H2S

In DNA, RNA, ATP and membranes PO43- is a source of phosporus

Trace Elements y y Inorganic elements required in small amounts Usually as enzyme cofactors

The Effect of Oxygen on Microbial Growth (Table 6.1) A. obligate aerobe- needs oxygen to survive B. facultative anaerobe- retain the ability to ferment to undergo anaerobic metabolism. Organisms not on top are facultative anaerobe. eg. E.Coli C. obligate anaerobe- cannot use molecule oxygen, but they can use oxygen from water eg. Plastrinium- causes tetanus and botulism D. Aerotolerant anaerobe-they don¶t use oxygen but can still grow in the presence of it. E. Microaerophile-can only grow in a very SPECIFIC level of oxygen

Toxic Oxygen Singlet oxygen: O2- boosted to a higher energy state Superoxide radicals O2y y y When you use oxygen as an electron acceptor Try to grab electrons from nearest molecule SOD (enzyme)* o O2-2+ O2-2 +2H+ Peroxide anion O2 y y y

* H2O2 +O2

catalase or peroxidase ± neutralize H2O2 catalase- produce H2O and O2 peroxidase- H2O

Hydroxyl radical (OH-) y y y Happens in cytoplasm Through ionizing radiation All of these could be toxic to humans or bacteria

Organic Growth Factors

-Organic compounds obtained from the environment. Varies from organism to organism -Vitamins, amino acids, purines, and pyramidines

Biofilms (Fig 6.5) -Microbial communities -form slime or hydrogels y bacteria attracted by chemicals via quorum sensing: communication between one cell and another. Drugs to block the quorum sensing receptors. y y y ³place gets too crowded, people have to leave.´ Important in digestive systems of cows. Need to digest cellulose. Grow upwards to increase surface area. Causes bacterial infections.

-share nutrients -sheltered from harmful factors - patients with indwelling catheters received contaminated heparin. -bacterial numbers in contaminated heparin were too low to cause infection -84-421 after exposure, patients developed pseudomonas infection -Pseudomonas fluorescens were cultured from catheters. -What happened? y y y They grew and multiplied by themselves. They are antibiotic resistance. Heparin might signal biofilm growth

Table 6.2: A chemically defined medium for growing a typical chemoheterotroph, such as E.Coli Table 6.4: Composition of Nutrient Agar, a Complex medium for the growth of heterotrophic bacteria Culture Media -Culture Media: nutrients prepared for microbial growth. -Sterile: no living microbes -Inoculum: introductions of microbes intro mediums -Culture: microbes growing in/on culture medium Growing a Culture -nutrients -pH -moisture -oxygen -initially sterile Agar -Complex polysaccharide

-Used as solidifying agent for culture media in Petri plates, slants, and deeps -Generally not metabolized by microbes -Liquefies at 100C -Solidifies at ~40C Culture Media (Table 6.2, 6.4) -Chemically defined media: exact chemical composition is known -complex media: you don¶t know the exact compositions. Include extracts and digests of yeasts, meats or plants y y Nutrient Broth Nutrient Agar

Anaerobic Culture Methods y y Anaerobic jar (fig. 6.6) Anaerobic chamber (fig. 6.8)

Reducing media -Contains chemicals (thioglycolate or oxyrase) that combine O2. -Heated to drive off O2. y Capnophiles -microbes that require high CO2 conditions -CO2 packet -Candle jar -multiple methods present Biosafety Level 1:No special precautions 2: Lab coats, gloves, eye protection 3: Biosafety cabinets to prevent airborne transmission 4: Sealed, negative pressure Selective Media -suppress unwanted microbes and encourage desired microbes. -chemically defined: salt media is preventing some bacterial species while allowing others to grow -Staphylococcus aureus- metabolizes manatose to make acidic environment (yellow) - Staphylococcus epidermidis- growing but basic Differential Media -make it easy to distinguish colonies of different microbes Enrichment Culture -encourages growth of desired microbe -assume a soil sample contains a few phenol-degrading bacteria and thousands of other bacteria Figure 6.6, 6.7

y y y y

inoculate phenol-containing culture medium with soil and incubate transfer 1 mL to another flask of the phenol medium, and incubate transfer 1 mL to another flask of the phenol medium and incubate only phenol-metabolizing bacteria will grow after much dilutions later.

Binary Fission (Fig 6.12A) Cell Division Bacterial Growth Curve

Obtaining Pure Cultures -A pure culture contains only one species or strain -A colony is a population of cells arising from a single cell or spore or from a group of attached cells. -A colony is often called a colony-forming unit (CFU) -The streak plate method is used to isolate pure cultures Preserving Bacterial Cultures -deep freezing: -50C to -95C -Lyophilization (freeze-drying): frozen (-54 to 72C) and dehydrated in a vacuum. The Growth of Bacterial Culture Reproduction in Prokaryotes y y y y y Binary fission Budding Conidiospores (actinomycetes)-chains when they reproduce Fragmentation of filaments Bacterial growth happens exponentially- based on generation number and binary fission can figure out how many cells are being made. Generation Time y If 100 cells growing for 5 hours produced 1,720,320 cells: o o Number of generation= [log #of cells (end) ±log #cells (beg)]/(0.301) log of 2 Generation time= (60min x 5hours)/ number of generations= 21 min/generation.

Bacterial Growth Curve Phases of Growth (log graph, not linear) 1. Lag phase -not much bacterial growth -no exponential growth 2. Log phase -exponential growth -assuming optimal conditions

-reaches point of capacity 3. Stationary phase -# cells growing = # cells dying -no exponential growth 4. Death phase -nutrients have been used up -exponential decrease Direct Measurement of Microbial Growth 1. Serial Dilutions 1:10 ± 9 mL broth in each tube 1:100 1:1000 1:10,000 1:100,000 2. Plate Counts a) pour plate method b) the spread plate method 3. Plate Counts y 4. After incubation, count colonies on plates that have 25-250 colonies (CFUs)

Counting Bacteria by Membrane Filtration Fig 6.18


Direct Microscopic Count -Serial dilution must be performed -number of bacteria/mL= #of cells counted/ volume of area counted


Turbidity Indirect Methods -turbidity -metabolic activity -dry weight

Direct Methods -plate counts -filtration -MPN -direct microscopic count

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The terminology of microbial control -Sepsis-microbial contamination -Asepsis- the absence of significant contamination -Aseptic surgery-prevent microbial contamination of wounds Sterilization- removing all microbial life Commercial sterilization- killing C. botulinum endospores Disinfection- removing pathogens Antisepsis- removing pathogens from living tissue Degerming- removing microbes from a limited area Sanitization- lowering microbial counts on eating utensils Biocide/germicide- kills microbes Bacteriostasis- inhibiting growth, NOT killing Population Death Rate is Constant A Microbial Death Curve Actions of Microbial Control Agents y y y Heat y y y y y y y y y Thermal death point (TDP)-lowest temperature at which all cells in a culture are killed in 10 min. Thermal death time (TDT)- time during which all cells in a culture are killed. Minutes to kill 90% of a population at a given temperature (fig 7.2) Moist heat denatures proteins Autoclave: steam under pressure Steam must contact item¶s surface Reduces spoilage organisms and pathogens Equivalent treatments 63 C for 30 min Decimal Reduction Time (DRT) Moist Heat Sterilization alternation of membrane permeability damage to proteins damage to nucleic acids (fig 7.1 A)

Steam Sterilization Pasteurization

y y y y

High temperature short time- 72C for 15 sec Ultra high temp- 140 C for <1 sec Thermoduric organisms survive Kills by oxidation o o o o Dry heat Flaming Incineration Hot-air sterilization Hot Air Equivalent treatments 170 C, 2 hr Autoclave 121 C, 15 min

Dry Heat Sterilization

Filtration y y y HEPA removes microbes > 0.3 uM Membrane filtration removes microbes > 0.22 uM Low temp inhibit microbial growth o o o y y y Radiation y Ionizing radiation (X rays, gamma rays, electron beams) o o y y Ionizes water to release OH Damages DNA Damages DNA Kill by heat; not especially antimicrobial Refrigeration Deep freezing Lyophilization

Physical Methods of Microbial Control

High pressure denatures proteins Dessication prevents metabolism Osmotic Pressure causes plasmolysis

Nonionizing radiation (UV, 260 nm) o Microwaves o

Principles of Effective Disinfection y y y y Concentration of disinfectant Organic matter Time pH

Use Dilution Test y y y Metal rings dipped in test bacteria are dried Dried cultures are placed in disinfectant for 10 minutes @ 20C. Rings are transferred to culture media to determine whether bacteria survived treatment.

Disk Diffusion Method Phenols and Phenolics y Bisphenols y Biguanides y Halogens y Iodine o o o y Tinctures: in aq. Solution Iodophors: in organic molecules Alter protein synthesis and membranes Bleach: HOCl (hypochlorous acid) Chloramine: chlorine + ammonia Chlorhexidine o Disrupt plasma membranes Hexacholorphene triclosan o Disrupt plasma membranes Disrupt plasma membranes

Chlorine o o

o Oxidizing agents

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TRANSCRIPTION-NUCLEUS Fig. 8.10 Eukaryotes: nucleus Prokaryotes: cytoplasm Fig 8.11 : RNA Processing in Eukaryotes DNA RNA transcript Exons- are expressed Introns- intervening sequences y y y Has to be removed In order to be translated for protein synthesis snRNPs mRNA

TRANSLATION- CYTOPLASM Fig 8.2 -mRNA is translated in codons (three nucleotides) -translation of mRNA begins at start codon: AUG (meth) -Translation ends at nonsense codons: UAA, UAG, UGA -64 sense codons on mRNA encode the 20 amino acids the genetic code is degenerate tRNA carries the complementary anticodon The Genetic Code (fig 8.8) y y y Note the degeneracy Cannot happen in eukaryotes DNA/RNA stuck in nucleus Simultaneous Transcription and Translation (fig 8.10)

The process of Translation (fig 8.9) AUG codon EPA sites New protein The Regulation of Gene Expression Regulation -Constitutive genes are expressed at fixed rate tRNA anticodon ribosomal units attaches

~60-80% of genes -Other genes are expressed only as needed y y y Operon Fig 8.12 DNA Regulatory gene Promoter* Operator* y y *Not genes: causes polymerase to bind Operator: acts as a stop light Repressible genes Inducible genes Catabolite repression

Structural Genes: Z Y A enzymes that ferments lactose (E. Coli) Regulatory gene: also a repressible protein (turns on AND off) Induction -Fig 8.12 -Lac operon when there is no lactose. -RNA polymerase binds at promoter, repressor binds the operator - Lac operon when there is lactose PRESENT y y Repression -Fig 8.13 -Structural genes: E D C BA y y encoding for different genes: tryptophan synthesis Repressor protein is ALWAYS on o Cannot bind at O, operator site Allolactose binds the repressor protein shape- no longer binds the operator. RNA polymerase can go ahead and make structural genes Z Y A

Once tryptophan is made y y y y Binds inactive repressor protein Blocking RNA polymerase Tryptophan: co repressor Allolactose: co- inducer

Catabolite Repression Fig 8.14a & b - Constitutively active genes -Preference energy source is GLUCOSE y growth rate is much faster than LACTOSE -When glucose runs out, cAMP builds up

y y y cAMP

cAMP= alarm mode Lag time Then, metabolizes LACTOSE

Lactose PRESENT, no GLUCOSE Inactive CAP Active CAP Promoter (extra red flag encouraging RNA polymerase to bind Fig. 8.15 Inactive CAP RNA polymerase cannot bind Inducer- allolactose there: promoting lactose metabolism) y y y y Mutation -A change in the genetic material -Mutations can be neutral, beneficial, or harmful y Neutral: Triplet code codes for the same amino acid o Non-gene encoding regions: neither beneficial or harmful LACTOSE +GLUCOSE Present

-Mutagen: agent that causes mutations -Spontaneous mutations: occurs in the absence of mutagen. Fig 8.17a, b,c,d Base substitution (point mutation)- change in one base y Sickle cell disease Missense mutation- Constitutively active genes result in change in amino acid Nonsense mutation- results in a nonsense codon. Peptide sequence cuts short, protein isn¶t being made. Frameshift mutation- insertion or deletion of one or more nucleotide pairs y y y y y Radiation Ionizing radiation (X rays and gamma rays) causes the formation of ions that can react with nucleotides and the deoxyribose-phospate backbone Huntington¶s Disease: accumulates in nerve cells Spontaneous mutation rate = 1 in 10^9 replicated base pairs or 1 in 10^6 replicated genes. Mutagens increase to 10^-5 or 10^-3 per replicated gene Nitrous Acid Nucleotide analog: looks like correct DNA base but they are NOT The Frequency of Mutation

Chemical Mutagen (fig 8.19a)

y y y y Selection*

Wave that breaks covalent bonds Photolyases separate thymine dimmers Nucleotide excision repair DNA polymerase makes the repair

UV Radiation and Repair (fig 8.20)

Positive (direct) selection detects mutant cells because they grow or appear different. Negative (indirect) selection detects mutant cells because they do not grow y y Fig 8.22 Amount of mutation that forms- increased incidents of cancer Media lacking histidine Incubation into Colonies of revertant bacteria y Salmonella + Rat liver extract Genetic Recombination -Vertical gene transfer: occurs during reproduction between generations of cells -horizontal gene transfer: Gene transfer between organisms without reproduction -Exchange of genes between two DNA molecules y y Fig 8.25 -Circular chromosome -DNA gets into the bacterial cell -One piece of DNA will replace the other -Result: Genetically transformed cell Fig 8.24 Griffith Experiment -Bacteria was encapsulated -Injected encapsulated bacteria into mouse dies. -injected nonencapsulated bacteria into mouse sick. -dead bacteria into mouse lives -Mixed bacteria mouse dies due to genetic transformation Fig 8.26 Bacterial Conjugation -One bacteria on one species can contact another species and can change genetic material Fig 8.27a.b,c Conjugation of E. Coli y F+ (has f factor), F-, Hfr (f+ recombined with chromosome) Crossing over occurs when two chromosomes break and rejoin Fig 8.23 Replica plating- (recorded) Fig 8.21- Looking for a auxotrophic mutation- being able to metabolize something

Ames Test for Chemical Carcinogens


Hfr + F-

Hfr cell + Recombinant F- cell

Fig 8.28 Transduction by a Bacteriophage -Usually talks about viruses -Transfers DNA from one host to another Plasmids Conjugative plasmid-carries genes for sex pilli and transfer of the plasmid Dissimilation plasmidsR factorsR factors, a Type of Plasmid (fig 8.29) y y y y y Origin of replication Similar map to bacterial chromosome- like a clock Segments of DNA that can move from one region of DNA to another Contain insertion sequences for cutting and resealing DNA (transposase) Complex transposons carry other genes

Transposons (Fig 8.30a b c)

Chapter 9
Biotechnology and Recombinant DNA y

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Biotechnology- use of microorganisms, cells, or cell components, to make product o Foods, antibiotics, vitamins, enzymes   Vector- self replicating DNA used to carry the desired gene to a new cell Clone- population of cells arising from one cell, each carries the new gene


Recombinant DNA o Insertion or modification of genes to produce desired proteins.

Fig 9.1 : A Typical Genetic Modification Procedure 1. Vector, such as plasmid is isolated (human insulin gene) 2. GO back into the bacteria 3. Make many copies y y OR Protein products of gene GMOs, or foods, hormones Interesting Facts Restriction Enzymes y PCR Inserting Foreign DNA into cell y DNA can be inserted into a cell by o o o o o o Obtaining DNA y y y Complementary DNA (cDNA) is made from mRNA by reverse transcriptase (fig 9.9) Synthetic DNA is made by a DNA synthesis machine Selecting a Clone (fig 9.11) o o o o o Putting obtained DNA into a special vector Vector inserted into bacteria Grown in a special plate Grows in certain color/ certain environment Obtain DNA of sequence Transformation Electroporation Protoplast fusion fig 9.5b Gene Gun Microinjection fig 9.6, 9.7 Cut specific

Table 9.2 : Used in pharmaceutical industry

Making a Product (vector for gene expression or the gene itself)


E.Coli o o Used because it is easily grown and its genomics are known Cells must be lysed to get product Used because it is easily grown and its genomics are known. May express eukaryotic genes easily May express eukaryotic genes easily Plants are easily grown May express eukaryotic genes easily Harder to grow


Sacchromyces cerevisiae o o


Plant cells and whole plants o o


Mammalian cells o o

Therapeutic Applications Human enzymes and other proteins Subunit vaccines Non pathogenic viruses carrying genes for pathogen¶s antigens as DNA vaccines Gene therapy to replace defective or missing genes RNA Interference (RNAi) y y y y Interference with RNA, cannot be used for translation Transcript control: cleaved. A way to gene silencing vs. gene knock out Nucleotides have been sequenced Human Proteome Project may provide diagnostics and treatments o Reverse genetics: block a gene to determine its function

The Human Genome Project

Scientific Applications y y y y y y Understanding DNA Sequencing organisms¶ genomes DNA fingerprinting for identification (fig 9.17) PCR Primers for a specific organism will cause amplification if that organism is present Reverse-trancription (RT-PCR): Reverse transcriptase makes DNA from viral RNA or mRNA. Real time PCR is different

Forensic Microbiology

Chapter 10: Classification
Taxonomy y y y y y The science of classifying organisms Provides universal names for organism Provides a reference for identifying organisms The study of evolutionary history of organisms All Species inventory (2001-2005) o o Placing Bacteria 1735 Kingdoms Plantae and Animalia 1857 Bacteria and fungi put in Kingdom, Plantae ± ³Flora´ 1866 Kingdom Protista proposed for bacteria, protozoa, algae, and fungi 1959 Kingdom Fungi To identify all species of life on Earth.

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Systematics, or Phylogeny

Identify which of those species might have diagnostic use and evolutionary history.

1961 Prokaryote defined as cell in which nucleoplasm is not surrounded by a nuclear membrane 1968 Kingdom Prokaryotae proposed 1978 Two types of prokaryotic cells (Bacteria and Archea) found- 3 Domain classification recommended y Fig 10.1/ Table 10.1 The 3-domain System (fig 10.1) Endosymbiotic Theory y y y y Pre-eukarya looked like prokaryote Plasma membrane invaginated DNA Determine common ancestors thru imprints of bones and shells Microorganisms are hard to find in fossilized form o o Phylogenetics y y Each species retains some characteristics of its ancestors Grouping organisms according to common properties implies that a group of organisms evolved from a common ancestor o o o Anatomy Fossils rRNA Black rock: Filamentous bacteria Upper right: cyanobacteria ~3 bil years ago

Fossilized Prokaryotes

Scientific Nomenclature y Common Names

o y

Vary with language and geography Used worldwide Escheria coli Homo sapiens- same all-knowing

Binomial Nomenclature (genus + specific epithet) o o o

Taxonomic Hierarchy Domain (bacteria is a domain not a Kingdom) y Kingdom* no kingdoms for bacteria o Phylum  Class Order y Family o Genus  The Taxonomic Hierarchy y y Fig 10.5 Appreciate Taxonomic Hierarchy- not inclusive Prokaryotic species: a population of cells with similar characteristics o o o Culture: grown in laboratory media Clone: Population of cells derived from a single cell Strain: genetically different cells within a clone. Same species, but specific subtype Classification of Prokaryotes Species

Phylogenetic Relationships of Prokaryotes y y Fig 10.6 Eukaryotic species: a group of closely related prganisms that breed among themselves o Animalia  o Plantae  o Fungi  o Protista  Classification of Viruses A catchall kingdom for eukaryotic organisms that do not fit other kingdoms Grouped into clades based on rRNA Chemoheterotrophic; unicellular OR multicellular, cell walls of chitin, develop from spores or hyphal fragments. Multicellular, cellulose cell walls, usually photoautotrophic Multicellular, no cell walls, chemoheterotrophic Classification of Eukaryotes


Viral species: population of viruses with similar characteristics that occupies a particular ecological niche. They do not fit in Domain, not living organism. Has its own classification system.

Classification and Identification y y Classification- placing organisms in groups of related species. Lists of characteristics of known organisms. Identification- Matching characteristics of ³unknown´ organisms to lists of known organisms. o o o y y y y Clinical lab ID Seen in hospitals Helps with classification ( p. 283 )

Dichotomous Keys Morphological characteristics- useful for identifying eukaryotes differential staining- gram staining, acid-fast staining biochemical tests- determines presence of bacterial enzymes

Identification Methods

Identifying a Gram- Negative, Oxidase ± Negative Rod (fig 10.8 ) Numerical Identification (fig 10.9) More Methods of Identification Serology- use of antibiotics for testing purposes y Aggluntination, ELISA, Western Blotting o Phage Typing y y Plate with lawn of bacteria. Each square of grid, add a different phage. Observe virus clearing of bacterial lawn- what type of viruses can infect this particular cell Based on how they are detected

Fatty Acid Profiles- FAME (fatty acid methyl ester) Flow Cytometry y y y Machine with microscopic probe. Solution sucked into a tube, passed into a laser picture. Take cell then use electrical conductivity- size, shape, density (more or less organelles, inclusions), cell surface Fluorescence- pick out one cell at a time DNA Base Composition- GC, AT ratio can be figured out. Indicative of relation. DNA Fingerprinting- RFLP¶s (chapter 9) restriction length polymorphisms are regions of DNA that has repeats in them. takes advantage of knowing DNA sequence. PCR- increasing frequency for amplification. Nucleic Acid Hybridization


Southern Blotting, DNA Chip, FISH

Building a Cladogram The Prokaryotes (check pic) Domain Bacteria y y Phylum Proteobacteria: o Class: o o o o o Alphaproteobacteria Betaproteobacteria Gammaproteobacteria Deltaproteobacteria Epsilonproteobacteria gram-negative, chemoheterotrophic

The Gammaproteobacteria y Pseudomonadales o Pseudomonas    y Vibrionales o Found in coastal water   y o Vibrio cholerae causes cholera V. parahaemolyticus causes gastroenteritis Opportunistic pathogens Metabolically diverse Polar flagella

Enterobacteriales (enterics) Petrichous flagella; facultatively anaerobic          Enterobacter Erwinia Escherichia Klebsiella Proteus Salmonella Serratia Shigella Yersinia

The Epsilonproteobacteria y y Slender gram-negative rods Helicobacter

o o o y

Multiple flagella Peptic ulcer Stomach cancer One polar flagella Gastroenteritis

Campylobacter o o

Nonproteobacteria Gram- Negative Bacteria y y Photosynthesizing bacteria Oxygenic Photosynthetic Bacteria: Phylum- Cyanobacteria o o y Gliding motility Fix nitrogen Purple sulfur, Purple nonsulfur: proteobacteria Green sulfur, Green nonsulfur

Anoxygenic Photosynthetic Bacteria o o

Phototropic y y y Oxygenic photosynthesis Anoxygenic photosynthesis Phylum: Firmiculates o o y Low G+ C Gram + High G+C Gram + Pleomorphic (many shapes) Actinomyces, Corynebacterium, Frankia, Gardnerella, Mycobacterium, Nocardia, Propionibacterium, Streptomyces o They look like protista but they are BACTERIA Nonproteobacteria Gram- Negatives y Planctomycetes o Gemmata obscuriglobus  y y Double internal membrane around DNA ( fig 11.23)

The Gram-Positive Bacteria

Phylum: Acitnobacteria o o o o

Life Cycle of the Chlamydias (fig 11.24a) Sprochetes- note axial filaments o o Borrelia Leptospira

o y


Fusobacteria o Fusobacterium    Are found in the mouth May be involved in dental disease Looks like toothpick

Domain Archaea y Extremophiles o Hyperthermophiles   o  o  Microbial Diversity y Bacteria size range o o y y Thiomargarita (750 uM) Nanobacteria (0.02uM) in rocks Pyrodictium Sulfolobus Methanobacterium Halobacterium

Methanogens Extreme halophiles

PCR indicates up to 10,000 bacteria per gram of soil Many bacteria have not been identified because they o o o o Haven¶t been cultured Need special nutrients Are a part of complex food chains requiring the products of other bacteria Need to be cultured to understand their metabolism and ecological role.

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Focus on Phyla- What distinguishes one phyla from another.

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