MICRO LECTURE NOTES EXAM #3

MICROBIAL GROWTH
TEMPERATURE: -Temperature plate an important roles in microbial like though its effects on the three dimensional configurations of biological molecules. -Different temperatures have different effects on the survival of microbes. -Mininmum temperature= no growth. -Optimum temperature= temperature that supports the fastest growth. -Maximum temperature=no growth. *FIG. 6.4

-Different temperatures have different effects on the growth of microbes. -Min. growth temperature= minimum temperature at which an organism is still able to conduct metabolism and grow. -Optimum growth temperature= temperature at which an organisms metabolic activities produce the most growth. -Maximum growth temperature= highest temp. that a microbe will grow. -at temperatures below the minimum growth temperatue of a microbe, proteins are rigid and growth is inhibited. -At low temperatures, lipids are rigid and hard. - At temperatures above a microbe maximum growth temperature, proteins are flexible and cannot maintain their 3-dimentional shape and so they denature. -at high temperatures, lipids melt. *FIG.6.5

-PSYCHROPHILES= grow within a range or -5 to 20C -Live in ice, snowfields, and cold water. -Some can cause food spoilage in refrigerated foods. -PSYCHOTOLERANT= minimum growth=0, optimum growth=25, maximum growth=35 -grow best at room temperature. -MESOPHILES= have a temperature range between 20-45C -grow at human body temp. - Mesophiles that grow between 20-30C are plants -Mesophiles that grow between 35-40 are organisms that prefer t grow in the body of warm-blooded hosts. -MESOPHILIC= min growth=15, optimum growth=37 (body temp.), maximum growth=45 -THERMOPHILES= grow at temperatures above 42C -THERMOPHILIC=min=42, optimum=65, max=80 -require higher temperatures. -No pathogens -have more saturated lipids in there membranes. -grow in habitats such as compost piles and hot springs. -HYPERTHERMOPHILES= have a min growh=65, optimum growth=90, maximum growth=105. -grow in water -EXTREME HYPERTHERMOPHILES= min growth=80, optimum growth=105 maximum growth=121 -only archaiae PH -organisms are sensitive to changes in pH because hydrogen ions and hydroxyl ions interfere with hydrogen bonding within protein and nucleic acids. -ACIDOPHILES= optimum growth= below 6 pH -mild growth= 6-4 pH (fungi) -extreme growth= below 4 pH (bacteria and archaea) -NEUTROPHILES= optimum growth at 6-8 pH -ALKALIPHILES= optimum growth= above 8pH -mild growth=8-10pH -extreme growth= above 10pH (archaea- also love salt.)

SALT

-most microbes like 0.7% -HALOTOLERANT= tolerant up to 7% -produce high concentrations of compatible solutes -HALOPHILIC=mild growth= 1.0-7%, moderate growth= 7-15%, extreme growth=15-30% OXGEN CONCENTRATION -Aerobes - STRICT AEROBES= grow best at 21% oxygen -MICROAEROPHILES= grow best at 12% oxygen - FACULTATIVE AEROBES= can grow with or without oxygen, but grow best with oxygen. -Anaerobes - STRICT ANAEROBES= cant grow with oxygen - AEROTOLERANT ANAEROBES= can grow in the presence of oxygen, but do not use the oxygen - OBLIGATE ANAEROBES= cannot grow in the presence of oxygen- oxygen is toxic to these. OXYGEN TOXICITY -superoxide and peroxide are the two main types of toxic chemicals -superoxide is the most toxic -Both are side products of aerobic respiration - they are produced in the environment by things such as metals. -Some organisms have enzymes that deal with toxic forms of oxygen. -CATALASE= converts hydrogen peroxide to water and molecular oxygen. H2O2 O2 + H2O -PEROXIDASE= breaks down hydrogen peroxide without forming oxygen H2O2 H2O -SUPEROXIDE DISMUTASE (SOD)= converts superoxide to peroxide -O2 radical -SUPEROXIDE REDUCTASE (SOR)= superoxide H2O

-Strict or facultative aerobes have catalase and SOD which allows them to deal with the toxins of their own metabolism. -Microaerophiles maintain lower concentrations of enzymes -Obligate and strict anaerobes have NO enzymes - Aerotolerant anaerobes have SOD *FIG. 6.3:

a. b. c. d.

have catalase and SOD only grow a bottom because oxygen is toxic to them grow best with oxygen but can grow with out it grows evenly because they grow without oxygen but they can tolerate it.

MICROBIAL DEATH RATE -Microbes don t die instantaneously; instead, they die at a constant rate over a period of time. *FIG. 9.1

CONTROLLING GROWTH IN THE ENVIRONMENT

-suffixes: -STASIS/STATIC= inhibition of growth, but not complete destruction of microbe

-CIDE/CIDAL= destruction of microbe -ANTISEPSIS= reduction of the number of microbes on living tissue -DISINFECTION= destruction of most microbes on non living tissues -ASEPTIC= refers to an environment or a procedure that is free of microbes -DEGERMING= removing of microbes by mechanical means (handwashing, alcohol swabbing at site of injection) -PASTEURIZATION= use of heat to destroy pathogens and to reduce number of spoilage microbes -SANITIZATION= removal of pathogens from objects to meet public health standards. -STERILIZATION=destruction of all microbes and viruses in or on an abject. -Both chemical and physical methods are used to control microbial growth in the environment. CHEMICAL METHODS OF CONTROL -DISINFECTANTS- controls growth on non-living things -ANTICEPTIC- controls growth on the surface of living things -both ways are non specific, meaning it effects lots of things-not just one. -They have non specific modes of action. -If a chemical effects a microbes proteins, it effects all the proteins the same way. -chemicals react with organic molecules. -Most chemicals result in protein denaturation. -different chemicals denature proteins in different ways. *FIG 9.15 -PHENOL- used both as a disinfectant and an antiseptic. - in early years it was used to clean surgery sites. -denatures proteins and dissolves lipids. -not used anymore -TRICLOSAN- found in lots of things such as soaps, and detergents. -dissolves lipids and denatures proteins. -ALCOHOL- used as a disinfectant, antiseptic and a degermer. -denatures proteins and disrupts cell membranes. -organic matter such a dirt interfere wit the alcohols ability to come into contact with the microbe (which is why using hand sanitizer on dirty hands doesn t get the hands as clean) -OXIDIZING AGENTS= used as a disinfectant and a antiseptic for deep wounds. -denatures proteins by oxidation. -used in deep wounds because the microbes in deep wounds are anaerobes. -SUFACTANTS (soaps and detergents)= saops=degermers, detergents=antiseptics. -surfactants are amphipatric, meaning it has both hydrophilic and hydrophobic components. -Hydrophilic components of the surfactants react with the hydrophobic components in the cells and the hydrophobic components of the sufactant work to dissolves the cells into water so that can be washed away.

-ALDEHYDES AND GASOUS AGENTS- chemicals that if used long enough (long contact) can achieve sterilization. PHYSICAL METHODS OF CONTROL -There are many ways to physically control growth in the environment: 1.Temerpature (hot and cold) 2.Filteration 3.Desication (removal of water) 4.Salt (osmotic effecs) 5.radiation TEMPERATURE: -cold=biostatic -heat=biocidal -The higher the temperature, the more quickly you can kill microbes -By increasing pressure in the environment, you can heat water to higher temperatures without boiling it. -There are two types of heat: dry and moist. MOIST HEAT -kills cells by denaturing proteins and destroying cytoplasmic membranes. -more effective then dry heat. -ca use lower temperatures for shorter periods of time to achieve the same results as dry heat. -boiling, pasteurization and heat under pressure are all examples of moist heat methods. -BOILING= disinfection/ sanitization to kill vegetative cells. -NOT a sterilizer -PASTEURIZATION= brief heat treatment designed to destroy pathogens and food spoilage microbes. -also used to increase shelf life of food. -HEAT UNDER PRESSURE= higher pressure allows temperatures to rise above 100C -AUTOCLAVE= sterilize equipment. -PRESSURE COOKER -DRY HEAT -absence of moisture. -requires higher temperatures for longer periods of time -used on things that are sensitive to moisture. -HOT AIR= sterilizes materials such as powders and oils by denaturing proteins, destroying membranes, and oxidizing metabolic compounds. -INCENERATION= oxidizes everything completely by applying the things to be incinerated at temperatures above 1000C -COLD TEMPERATURES (FREEZING/REFRIGERATING= inhibits metabolism -DESICATION/DRYING= biostatic process that involves the removal of water. -FREEZ DRYING (LYOPHILIZATION)= used to preserve cultures -OSMOTIC EFFECTS= use of high concentrations of salt in foods to draw water out of cells in order to inhibit microbial growth. (biostatic process)

-FILTRATION= using a filter to pass air or liquid through in order to physically separate microbes from the air or liquid. -RADIATION= two types: ionizing and nonionizing. -IONIZING RADIATION=denature DNA, causing fatal mutation of cells. -electron beams, x-rays, gamma rays -NONIONIZING RADIATION= damages DNA in cells. -UV light -longer wavelengths. -involves the formation of thymine dimers which inhibit DNA transcription and replication -The shorter the wavelength, the more damaging.

CONTROLLING GROWTH IN THE BODY

-Chemical use to control microbial growth in the body is SELECTIVE (targets the pathogen and not the host) -ANTIMICROBIAL AGENTS=target microbes that cause infections. -include antibacterials, antivirals, antifungals, and antibiotics. ANTIBIOTICS: -means anti-living -Drugs that inhibit or kill infectious agents. -They don t kill viruses. -antibiotics are chemicals produces by fungi or bacteria that target other bacteria. -SELECTIVE TOXICITY= ability to target specific bacteria and not effect the host. -antibiotics have 5 modes of action that can explain how they target bacterial cells. -MODE OF ACTION=how it works 5 MODES OF ACTION OF ANTIBIOTICS: 1.Inhibit cell wall synthesis -these drugs are selectively toxic to certain fungal or bacterial cells, which have cell walls, but not to animals, which lack cell walls. -target peptidoglycan layer in cell wall. -inhibit peptifoglycan formation by binding to enzymes that cross-link the NAM subunits. -With incorrectly formed peptidogycan layers, bacterial cells have weakened cell walls and are less resistant to the effects of osmotic pressure. -effective only on bacterial cells that are growing or reproducing because these drugs only prevent the formation of pept. and have no effect on preexisting peptidoglycan layers. This dormant cells are unaffected. 2.Inhibit protein synthesis -drugs that inhibit protein synthesis (translation) do so by targeting the differences between prokaryotic and eukaryotic ribosomes. -they selectively target the 70S ribosomes of prokaryotic cells which are composed of 30S and 50S subunits.

-the 30S and 50S subunits play a role in the initiation of protein synthesis, in codon recognition, and in the docking of tRNA-amino acid complexes. -Some drugs can change the shape of the 30S subunit, making it impossible for the ribosome to read the codons of mRNA correctly. -TETRACYLINES block the tRNA docking sites , which then prevents the incorporation of additional amino acids into a growing polypeptide.(this inhibits the growth of the protein) 3.Inhibit nucleic acid synthesis -drugs function to target the nucleic acids DNA and RNA -Block either the replication of DNA or the transcription into RNA. -interfere with the ability og nucleic acids to bind together to form chains. -some bind to enzyme active sites in order to block enzymes from binding, thus, inhibiting nucleic acid synthesis. 4.Interfere with cell membranes -some drugs target lipids in the membranes -prevent lipid synthesis or attach to specific lipids that are found in the bacterial membranes. -disrupts membranes causing lysis of the ell. -disrupts transport across membranes. -block lipid synthesis when then drug binds to a specific enzyme used for lipid synthesis. 5.Interfere with a metabolism -many bacteria make their own folic acid which is a man type of metabolic pathway. -some drugs interfere with folic acid synthesis -PABA is crucial for the synthesis of nucleotides (folic acid) required for DNA and RNA synthesis. -PABA converts to hihydrofolic acid then to tetrahydrofoloic acid which is used for DNA synthesis. -SPECTRUM OF ACTIVITY= characteristic of antibiotic that refers to what the antibiotic acts on -BROAD SPECTRUM=effect many types of bacteria -NARROW SPECTRUM= effects only a few types.

RESISTANCE TO ANTIBIOTICS
-specifically to antibacterial drugs -bacteria is resistant to drugs not to hosts -bacteria have their own modes of action they use to resist that modes of action of antibiotics 5 MODES OF ACTION OF RESISTANCE 1.Some resistance cells may produce their own enzymes that destroy or deactivate a drug. -ex: some produce an enzyme called B-lactamase which breaks the drug. 2.Modify the target of the drug so that the drug no longer works properly

-by modifying the target of the drug, the drug can no longer attach to that target, or it binds to it less effectively. 3. Slow or block the entry of a drug -typicaly envolves changes in structure or electrical charge of their cytoplasmic membrane proteins that constitute channel pores. 4. Alter their metabolic chemistry, or abandon the metabolic step altogether. -this makes it so the drug can no longer effect the bacteria s metabolism. 5. Pump the antimicrobial out of the cell quickly before the drug can act. -EFFLUX PUMP (resistance pump)= pump powered by ATP that pumps out drugs. -pumps are important because they can pump more then one type of drug out and most of them are on plasmids that can be shared between bacteria. *FIG 10.2 and FIG 10.6

-TRANSFORMATION= When chromosomal DNA that is lost from a dead lysed cell is taken into a nighbor cell and broken down and incorperated into that live cell s DNA -When this happends the live cell is not transformed because it now has the resistance that the lysed cell had. -TRANSDUCTION= involves the transfer of DNA from one cell to another via replicating virus called a PHAGE -the phage injects its genome into the host cell and directs it to synthesize new phages -CONJUGATION=The physical contact between a donor cell and a recipient cell through PILI that takes place in order to transfer genetics in bacteria. -unlike in transformation and transduction methods, the donor cell in conjugation remains alive. -all chromosomal information is picked up via vertical of horizontal gene transformation. -VERTICAL GENE TRANSFORMATION= the passing of genes to the next generation. -HORIZONTAL GENE TRANSFORMATION= the passing of a microbes genes to another microbe of the SAME generation. -transduction, translation, and conjugation are types of horizontal gene transformation. -involves a donor cell passing on part of its genome to a recipient cell.

MICROBIAL GENETICS