1) Define the Human Microbiota - The Human Microbiota consists of all of the bacteria that are associated with

the human body - Relationship between Microbiota and Host is Mutualistic and/or Symbiotic in that both the host and microbiota benefit from the partnership. - Consists of microbial cells from all three kingdoms: Bacteria, Archaea and Eukarya - The number of microbial cells to human cells is ~ 10:1 - Microbial populations are distinct for each niche within the body (ie. Intestines houses a different population than on the skin) - Every individual may be unique in their Microbiota - Microbiota communicates with and can trigger responses in host - Microbiota is required for prostnatal development of systems (immune system and GI) - Microbiota communicate among themselves to promote a cooperative response - Supplies Host with Vitamins and other Cofactors - Destroys toxic agents - Digests substrates (polysaccharides) which the host cannot digest - Protects host by excluding certain disease causing (pathogenic) virulent microbes 2) Describe the Morphology and Anatomy of Bacteria - Bacteria are Prokaryotes and therefore do not have organelles (No mitochondria, No Nuclear Membrane, No Chorloplasts and No ER), which are found in Eukaryotes - Some bacteria are capable of Photosynthesis, Autotrophy (eat non-carbon materials), Sexual Mating and Communicating with each other. - Mitochondria and Chloroplasts developed from bacteria; Likely that Mitochondria developed from Rickettsia prowazekii. - Usual size is 0.2 to 80 um in length in Rods, and 0.5 um diameter for cocci - Cytoplasm is usually packed, causing an increased intracellular pressure, equivalent to ~4 atm. - 70% of cytoplasm is water - Cytoplasm also contains storage areas for lipids, carbohydrates and other essential nutrients Bacterial DNA - A Bacterial Genome consists of all DNA within the cell, this includes extrachromasomal plasmids, bacteriophage DNA or RNA and Episomes. - Most bacteria have a single, circular chromosome composed of a DNA duplex, which is folded and coiled and about 2mm in length (~1000x cell length). - Haploid - No Histone proteins. DNA structure is maintained by Polyamines such as Spermine and Spermidine. - Some bacteria have multiple linear chromosomes: Borrelia burgdorferi, Brucella abortus - Plasmids are Extrachromosomal that are small (smaller than chromosome) and usually circular, but can be linear. Plasmids are self-replicating. Morphology - Coccus = Spherical - Bacillus = Rod - Spirillum = Spiral shaped - Coccobacillus = Oval (mix of Coccus and Bacillus) - Fusiform Bacillus = stretched out oval - Virbrio = Boomarang shaped

Attached to Peptidoglycan and anchored into cell membrane by fatty acids .Gram – Only ..Lysozyme is a natural enzyme in mammals that breaks the Beta-1.Acts as a Permeability barrier and confers innate antibiotic resistance.may be used to attach some bacteria to specific hosts surfaces Outer Membrane . Improper formation of Peptidoglycan layer is lethal to bacteria.The multiple layers of Peptidoglycan are cross-linked with peptide bridges . Glutamic Acid and Lysine or Diaminopimelic Acid.Antigenic – activates complement .Found in Gram + cells only . This Tail is used to link with Nacetylmuramic Acid sugars of other layers in the Peptidoglycan .Consist of Phosphodiester polymers of Ribitol or Glycerol with linked D-amino acids and sugars . . the Cell Membrane Gram – cells stain Pink in Gram Stain (due to Gram Saffranin) Gram – Cells have a Thin Peptidoglycan layer Gram – Cells have two Lipid membrane.Peptidoglycan is made up of a Dissaccharide: N-acetylglucosamin and Nacetylmuramic Acid . This space contains the thin peptidoglycan layer and also contains specialized enzymes not found in Gram + Cells Note – Mycoplasma do not contain a Peptidoglycan layer Peptidoglycan (Cell Wall) .4 link in the Peptidoglycan Dissaccharide Teichoic Acids .N-acetylmuramic Acid contains a Cross-Linking Tail.Spirochete = Corkscrew shaped Gram Positive vs Gram Negative Gram + cells stain purple in Gram Stain (due to Crystal Violet) Gram + cells have a Thick Peptidoglycan layer Gram + cells have only one lipid bilayer. consisting of Alanine. . Therefore. the Cell Membrane which is a lipid bilayer and an Outer Membrane which contains a single Lipid Layer facing inside the cell and a Lipopolysaccharide layer facing towards the exterior of the cell Gram + cells contain Teichoic Acid on their Gram – Cells contain Lipopolysaccharide on outer layer of Peptidoglycan the exterior portion of their Outer Membrane Gram + cells have No Periplasmic space Gram – Cells have a Periplasmic space between the Inner Cell Membrane and the Outer Membrane. the Peptidoglycan synthesis system is the target of many Antibiotics.Peptidoglycan protects the cell and prevents osmotic rupture of the cell and also provides the cell with its characteristic shape. and protects the cell from Lysozyme and Complement attack.

Phospholipid Bilayer that does not contain any Cholesterol or Sterols (Bacteria do not make Sterols) . Also used to Identify Gram Negative Organisms (ex.Used to Deliver Toxins 2) Sex Pili (conjugal) – Male cell has one or two conjugal pili that locates receptors on female cells . such as the lungs. Detergents. .When a Male Sex Pili binds to a Female cell. Ribosomes and Protein Synthesis.Allows for Surface Motility .LPS consists of 3 domains: 1) O-antigen – Saccharide repeats. These normal floral biofilms prevent entrence of pathogens. . However flagella are not found on all bacteria.Allow for immune system escape by altering the pilin antigenic type . consisting of 1 to 40 repeats. DNA Replication. Appendages . which form a hollow channel. . intestines and vagine are covered with normal flora biofilm embedded in their capsules. Bacillus anthracis) .Flagellum function in a cork-screw like manner propelling the bacterium . Chemotaxis.Porins are Trimeric Proteins that can open and close to permit passage of solutes . oropharynx.Contains the enzymes for Electron Transport.Usually composed of a Polysaccharide material.Poorly antigenic.Some may be composed of a Polypeptide (ex. Antibiotics and Antibodies . Consists of Protein Subunits called Pilin. the Sex Pili will undergo Depolymerization.May help a bacterium stick to surfaces due to Adhesins at the ends of the pili.Inhibits Complement attachment . Two types of Pili: 1) Somatic Pili – a single cell may consist of hundreds of these. The outer most part of the LPS which extends out from the cell.Flagellum – Used by many bacteria for motility.Contains Lipopolysaccharide (LPS) on the outer surface. also called a Glycocalyx . .Contains Porin Channels and Transport Proteins that take up nutrients . reeling in the female cell and allowing for DNA transfer to the Female.Allows the bacterium to stick to surfaces as well as helps in the formation of Biofilms . Lipid A attaches the Core-protein and O-antigen to the Outer Membrane Cytoplasmic Membrane . Destruction of this barrier can lead to disease. Lipid Synthesis. and shorten. and protects the cell from Phagocytosis.Pili (Fimbriae) – found on some bacteria. etc… Capsules .. E. .coli 0157:H7 serotype has a unique O-antigen) 2) Core protein – consists of an inner and outer core protein 3) Lipid A – acts as an Endotoxin when released from the cell during lysis.Most exposed host surfaces.

Mycobacterium contain a Arabinogalactan layer between the Mycolic Acid layer and Peptidoglycan Archea Cell Walls . has been found to have an Outer Membrane. These are called Axial Filaments. Anaerobic and Aerotolerant (Facultative anaerobes) . Mo.Makes the Mycobacterium more resistant to disinfectants and drying out than other bacteria (except spores are more resistant).Strongly resembles a Type III secretion System. Structure and function is almost identical to the ATPase. .Nitrogen fixation from the atmosphere is a process that is exclusive to bacteria. K. which is used to inject Toxins into the host cells. . only a single Archea.Membrane-associated transport systems concentrate solutes. Co. but rather spiral around the bacterium. the Flagellum produces new Flagellar Protein Subunits .Stained with Acid-Fast staining.Found on specific bacteria – Mycobacterium (which includes M. tuberculosis) . Sulfate. it spins the flagellum. both organic and inorganic (metals and ions). and impart a Screw-Like motility to the bacterium which allows it to penetrate viscous materials. .Flagellum is propelled by a flow of ions – either protons or sodium. Rather. . Mycolic Acids – A lipid material containing 60-90 carbons bound to the cell surface. Mg) and trace elements (Fe. .Oxygen requirements can differ between Aerobic. . changing rotation direction allows the bacterium to turn and is the basis for Chemotaxis . meaning they oxidize organic compounds for energy and obtain carbon from organic compounds.All bacteria associated with human and animal disease are Heterotrophic. they release enzymes into the surrounding area that digest food particles. or Peritricous (all around the cell) .All heterotrophic organisms require water and some inorganic compounds (Phosphate. others can fix nitrogen from the atmosphere (N2  NH3) . 3) Describe the Growth of Bacteria in a Host .Currently. such as mucus.The Archea have a surface layer of proteins . which can then be transported into the cell. Zn. without it life on earth would end. . Bipolar (on both ends).Rotates clockwise or counterclockwise. Instead of injecting toxins. in the cytoplasm.Some obtain Nitrogen from inorganic salts. .Spirochetes contain specialized Flagellum which do not extend out of the cell.Archea do not contain Peptidoglycan . except it doesn’t produce ATP. Mn.May be arranged on the cell in Polar (at end of cell). ..Mobility / Vigor of motility of bacteria can depend on the number of flagellum present . barrel shaped components with external receptors that are highly specific for the solutes they transport.Contain L-Glycerol instead of D-Glycerol which is found in bacteria . Ignicoccus. Cu) .Bacteria do not engulf/phagocytose food particles.Transport systems include membrane-spanning.

. Hyperthermophiles (>100 d. .The Electron Transport Chain is located in the Cytoplasmic Membrane. cells can do Fermentation to regenerate NAD by converting Pyruvate into Lactic Acid. Gyrase (Topoisomerase). Patients with Gaseous Gangrene can have bubbles of H2 under their skin that can be palpated.Some marine bacteria use a Sodium (Na) flow to turn the flagellum. . C). .5 mm and is therefore folded to fit in a 1 mm diameter cell . . They cytochromes and have no electron transport system.Flagella use the same mechanism to turn. . Thermophiles (45-80 d.Lactic Acid fermentation is the sole energy providing process in many Streptococci and is the likely cause of Disease by Streptococcus mutans. therefore.Obligate Anaerobes – Fermentation only or Anaerobic Respiration . tetani. .Anaerobic species that are capable of doing Oxidative Phosphorylation use something other than Oxygen as the terminal electron acceptor of the Electron Transport Chain. do Substrate Level Phosphorylation (Fermentation) and/or Oxidative Phosphorylation.Temperatures which bacteria can withstand range from Mesophiles (35-44 degrees C). the Pyruvate produced in the Glycolitic cycle can be processed into FADH2.Clostridium sp. . .Aerotolerant Forms – Fermentation.Substrate Level Phosphorylation – occurs with the conversion of Hexose to Pyruvate by taking a Phosphate from ATP and placing it on to the Hexose. therefore Protons are pumped out of the cell.Those that grow well in humans are Mesophiles. where Protons are pumped out of the cell creating an electric gradient. C).Most members of Genus Streptococcus fit the Aerotolerant category. The concentration of NAD is limited in a cell. this process uses a NAD and converts it to NADH. which are then excreted.Obligate Aerobes – Aerobic Respiration .The circumference of DNA is about 1. DNA Replication .No visible condensation of DNA . they turn the flagellum. creating an electric potential. Alcohols and other producs. Specifically. Protons flow back into the cell through an ATPase (ATP motor) which generates ATP for the bacterium. . . C). but often tolerate low atmospheric oxygen levels do not produce any . which causes dental caries.Those species that are aerobic and are in the presence of Oxygen can use the NADH in the Electron Transport Chain. Commonly NO3. Psychrophiles (15-20 d.Replication of the DNA begins at the Ori-C (Origin of Replication). GTP and CO2 in the TCA cycle. CO2. . FADH2 can be used in the Electron Transport Chain also. those that get energy from organic compounds. Vibrio cholera uses a sodium driven flagellum in the environment. and involves Helicase. (botulinum.is oxidized to NO2-. however when its infected a human it uses a different set of flagella that requires Protons. As Protons flow back in.Facultative Forms – Fermentation or Respiration . It takes 256 Protons to turn the flagellum 360 degrees. and produce various products. including H2 which is the gas in Gaseous Gangrene. Primase and DNA-dependent DNA Polymerases. and perfringens) also use Fermentation to produce energy.Heterotrophs. . Furthermore.

to transport Na.Topoisomerases are the target for Quinolone Antibiotics. producing a Minicell which contains no DNA. Biofilms are formed by attachment to biotic or abiotic surfaces by Pili. LPS and other receptors.Joint prostheses . . A biofilm is a microcosm of surface attached cells enclosed by a capsular matrix usually composed of polysaccharide.The Stationary phase occurs when nutrients and space become limited and competition ensues. Ca++ out of the cell.The most preferred method of growth is in a biofilm. .Replication is Semi-conservative and Bidirectional and rapidly growing cells may have multiple replication forks . Transport (Uptake) Systems .Leading Replication Strand will be in the 5’3’ direction .Energy sources for transport come from the Proton Motive Force. Mutations in the Min proteins result in the off-centering of the Z-ring and uneven separation of the cell during division.Cell division occurs soon after replication of DNA is complete Cell Division . in a manner of seconds. Min proteins form a coiled structure which cycles back and forth from both ends of the cell. .Glue that binds struvite kidney stones . Death rate of cells equals growth rate .Viscous clumps of Mycobacterium tuberculosis . glycocalyx. Gene Expression is different with cells in biofilms compared to .Cells contain numerous uptake processes.Bacterial Iron Transport systems are key Virulence Factors in many bacteria Bacterial Growth Curve (Planktonic Growth) . Proline into the cell… .Following the Lag phase is the Log phase. where bacterial growth does not occur and bacteria are in the process of upregulating the appropriate enzymes for growth. The Z-ring pinches off the cell into two equal cells. ATP hydrolysis and Phosphate in Phosphoenolpyruvate -Ex.Transport systems may be very specific for the solute which they transport . and in doing so locates the center of the cell.Many human diseases may involve Biofilms: . A protein called the Min Protein is responsible for locating the center of the cell. . where growth is exponential .Death phase occurs when nutrients become scarce and cannot support the bacteria.Cell division occurs by the formation of a Z-ring at the center of the cell by the FtsZ protein (a tubulin homolog)..Cells of biofilm are physiologically different from the planktonic (freefloating) cells.Dental plaque and other oral biofilms .Pseudomonas aeruginosa – Cystic Fibrosis of lungs .Lagging Strand is in the 3’ 5’ direction . Antiporters and Symporters can use Protons that are pumped out (part of the proton motive force).Haemophilus influenza – ear canal infections .Legionella pneumophila clumps wafted from air conditioners .Consists of a Lag phase. .

.Spores can survive for an unknown length of time and have essentially no metabolic activity. The Coat(s) layer is impervious to many chemicals.Spores can be killed by Formaldehyde. but will begin germination almost immediately in favorable conditions. but NO mRNA). with no similarity to the peptidoglycan layer of a cell. ionizing radiation. cereus) . Ethylene oxide or Steam under pressure (autoclave) .Senescent / non-dividing cells are able to escape antibiotic treatments because most antibiotics require cells to be actively dividing for them to have an effect. Below the Coat(s) layer is a Peptidoglycan layer and then a Inner membrane that resembles the Cytoplasmic membrane. . protein synthesis apparatus (tRNA and ribosomes. enzymes for amino acid synthesis.planktonic. B. Coat(s) layer which consists of Keratin-like protein with disulfide bonds. The cells release a Acyl-HSL signal which begins the formation of the biofilm .Biofilms have specialized architecture that allows for water channels .Polysaccharide matrix of a biofilm provides protection from UV and chemicals (antibodies.90% of biofilms on prostetic devices consist of Staphylococcus aureus and Staphylococcus epidermidis 4) Relevance to Disease . Survivors of the antibiotic treatment were in a non-dividing state during the treatment and are called Persisters.Endospores allows bacteria to persist in a dormant state in a very protective capsule . UV.Biofilms form from Planktonic cells that adhesins and are brought together by some environmental signal. Endospore structure consists of a Exosporum which is the remaining membrane of the cell after formation of the endospore. .Spores are resistant to commonly used disinfectants and antiseptics. . Calcium. energy producing enzymes. antibiotics).Biofilms may be beneficial to host by excluding incoming pathogens. Persisters that survive antibiotic treatment ARE NOT resistant to the antibiotic. . prevents dessication and osmotic shock. Different types of bacteria may work together in a biofilm and may exchange genetic material with each other. anthracis.A fraction of a bacterial population will survive a stress like an antibiotic. DPA and some small pore proteins cross-link to provide protection from heat. desiccation.Infection with a virulent bacterium that overcomes exclusion by the microbiota and host-related defenses.Immunosuppression.The core of the endospore contains DNA. . either drug induced or disease induced . .Biofilms are organized communities of bacteria .Biofilm growth provides protection from antibiotics and antibodies. .Antibiotic chemotherapy which damages select portions of the microbiota . 5) How do pathogenic bacteria overcome host defenses? . Dipicolinic Acid (DPA). Bacterial Spores – produced by two gram-positive genera: Bacillus (pathogens are B. chemicals and radiation. essential for development of GI system and Immune system.Disruption or imbalance of the host-microbiota can be caused by several conditions: .

. wastes and deleterious agents like antibiotics .ex. toxins. usually sterile.Siderophore mediated iron transport – Siderophores are iron chelating (binding) molecules that are excreted by the bacterium to capture iron.Secretion systems can be virulence factors . How do microbes cause disease? . There are atleast six (numbered I through VI) distinct secretion processes.Type III systems are found only in pathogens and are used to pump toxins and virulence factors.Secretion systems release digestive enzymes. . Ferrous Iron Transport . . Secretion Systems .Bacterial Iron Transport Systems are key virulence factors in many bacteria .Type I systems rapidly excrete Antibiotics.Genes that confer the Virulence phenotype on a bacterium occur on both chromosomes and plasmids. anatomic site. some plasmids harbor genes required for virulence.Overgrowth of one member of the microbiota .In Gram Negative bacteria. sometimes rapidly enough to confer resistance .Persistance of mutants – some cells may acquire spontaneous mutations that allow them to be more resistant to the bodies defenses or antibiotics 6) What are the virulence mechanisms of pathogenic bacteria? . secreted substances must pass through two membranes.Transfer of microbiotal cells to the “Wrong”. In short. Heme Transport. Iron Transport .Disruption or imbalance of the host-microbiota relationship may cause infectious disease by: . DNA.Vulnerability to incoming pathogens . sometimes directly into the host.

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