Microbiology Exam II Review Chapter 4: Bacterial Culture, Growth and Development 4.

1 Microbial Nutrition Nutrient Supplies Limit Microbial Growth Essential Nutrients-are those compounds a microbe cannot make itself but must gather from its environment if the cell is to grow and divide Macronutrients-nutrients needed in large quantities. CHONSP-make up the carbs, nucleic acids and proteins of the cell. Mg2+, K, and Fe2+ act as regualtory cofactors to signal molecules Ca2+ Micronutrients-are ubiquitus contaminants-required because these elements are frequentlyessential components of enzymes or can themselves be components of cofactors Microbes Evolved to Grow in Different Environments growth factors-specific nutrients that are not required by all cells defined minimal medium contains only those compounds needed for an organism to grow. 99.9% of all the microorganisms in earth cannot be grown in the laboratory at all! A DNa amplifying procedure called Polymerase chain reaction PCR can be usedto screen for the presence of these genes in the soil and water samples. Microbes Build Biomass through Autotrophy or Heterotrophy The carbon cycle, a critical part of maintainig life and recycling key nutrients involves two counterbalancing metabolic groups of organisms: heterotrophs and autotrophs. Heterotrophs (like E. Coli) rely on other organisms to form organic compounds, such as glucose, that they use as carbon sources. During heterotrophic metabolism, organic carbon sources are disassembled to generate energy and then reassembled to make cell constituents. This process converts a large amount of the organic carbon source into CO2, which is them released to the atmosphere. photoheterotrophy-produces energy through photolysis of organic compounds. Organic compounds are broken down and used to build biomass Chemoheterotrophy (organotrophy or heterotrophy)-yields energy and carbon for biomass solely from organic compounds. Autotrophs fix CO2 as a carbon source, reducing ot (adding Hatoms) to male complex cell constituents made up of CHO. These organic compounds can later be used as carbon sources by heterotrophs photoautotrophs-use light for photosynthesis photoautotrophy-generates energy through light absorption by the photosynthesis (light-activated breakdown) of H2O or H2S. This energy is used to fix CO2 into biomass

chemoautotrophy- (lithotrophy) produces energy from oxydizing inorganic molecules such as Fe, S or N. This energy is also used to fix CO2 into biomass note that many species can utilize more than one of these strategies, depending on environmental conditions and whose products carry out different functions. note Eukayotes carry out only a limited range of heterotrophic and photosynthetic reactions

"-trophy" the energy yielding metabolism: Carbon Source for Biomass Auto- CO2 is fixed and assembled into organic molecules Hetero-preformed organic molecules are aquired from outside and asembled Energy Source Photo- light absorption exites electron to high-energy state Chemo-chemical electron donors are oxidized Electron source Litho- inorganic molecules donate electrons Organo-organic molecules donate electrons 4.3 Culturing Bacteria Bacteria Are Grown in Culture Media Pure Culture-single strain of a single speciesLiquid media is convenient for examining growth kinetics and microbial biochemistry at different phases of growth dilution streaking- technique when a drop of liquid is collected using an inoculating loop and streaked across the agar plate surface in a pattern spread plate technique-starting from liquid culture, a series of tenfold dilutions placed directly on the surface of separate agar plates and then spread out using a sterilized, bent glass-rod. Growth Complex in Complex Media or in Synthetic Media synthetic media-nutrient rich but porly defined complex complex media-precisely defined media enriched medium-fastidious, e.g. requiring that components of blood be added to basic complex medium differential media- expose biochemical differences between two species that grow equally well 4.4 Counting Bacteria Coulter counter- a microbial culture is forced through a small orifice, through which flows an electrical current. Electrodes placed on both sides measure resistance. Every time a cell passes through the orifice, electrical resistance increases, and the cell is counted. Works best with larger

eukaryotic cells, such as red blood cells, the instrument is not generally sensitive to individual bacteria Viable Cells Estimate the Number of Cells That Can Form Colonies Pour plate-agar subsequently poured into an empty petri plate, where the agar cools further and solidifies viable bacterial organism-defined as being capable of replicating and forming a colony on a solid medium surface 4.5 The Growth Cycle binary fission-one parent cell splits into two equal daughter cells, note that some species divide assymetrically Unlimited Population Growth is Exponential growth rate- rate of increase in population numbers or biomass, is proportional to the population size at a given time. Such a growth rate is called exponential because it generates an exponential curve, a curve whose slope increases continually. 2^n, where the exponent n represents the number of generations that have taken place between two time points A constant Generation Time Results in Logarithmic Growth generation time-length of time from one generation to the next--the constant interval at unlimited resources. Varies with bacterial species, type of medium, temperature and pH. Also known as the doubling time because the population doubles over one generation. number of cells present: N0 x 2^n example: 1 cell x 2^3 cells= 8 cells Exponential growth occurs only for a short period when all nutrients are in full supply and the concentration of waste products has not become a limiting factor. The steeper the slope, the faster the organisms are dividing The mathematics of exponential growth apply at least until the community reaches a density at which different species begin to compete Stages of Growth in Batch Culture The simplest way to model the effects of changing conditions is to culture bacteria in a liquid medium within a closed system, such as a flask or tube. In this batch culture, no fresh medium is added during incubation'; thus nutrient levels decline and waste products accumulate during growth As medium conditions deteriorate, alterations occur in membrane composition, cell size and metabolic pathway, all of which impact generation time. Lag phase-lag period, where cells do not divide.

Several reasons- cells may have taken from an aged culture may be damaged and require time for repair. Also dependent on pH, temperature, type of medium. Early log or exponential phase- it's balanced growth, where all cell components are synthesized at constant rates relative to each other.

Growing and dividing at the maximum rate possible based on medium and growth conditions. Cells are largest at this stage. Failure to adjust to this stage will lead to increased mistakes in RNA, protein and DNA synthesis Late Log Phase- the rate of doubling eventually slows, and a new set of growth-phase-dependent genes are expressed

at this point, some species can also begin to detect the presence of others by sending and receiving chemical signals in a process known as quorum sensing Stationary Phase- when the growth curve levels off

caused by lack of key nutrients or buildup of waste products happens at about 10^9 cells per milliliter Eukaryotes enter at lower numbers because they are bigger and run out of nutrients sooner Death Phase: death rate-the rate at which cells die (logarithmic)

difficult to define because mutations arise that promote survival. Consequently, the death phase is extremely prolonged Continuous Culture Maintains Constant Cell Mass during Exponential Growth In a continuous culture, fresh medium is continually added to a culture and an equal amount of culture is constantly siphoned off

bacterial populations can be maintained at a constant cell mass for an extended periods of time cells achieve a steady rate, which permits detailed analysis of microbial physiology at different growth rates cell mass eventually decreases in the vessel , this is known as washout

4.6 Biofilms Biofilms-specialized, surface, attached communities Biofilms-the specific environmental signal induces a genetic in planktonic cells. The planktonic cells then start to attach to nearby innanimate surfaces by means of flagella, pilli, lipopolysaccharides or other appendatges and begin to coat the surface with an organic monolayer of polysaccharides and glycoporoteins to which more planktonic cells can attach

quorum sensing, cells begin to communicate with each other by sending and receiving signals. These chemical signal molecules are continually made by individual cells once the population reaches a certain number 'quorum' the chemical signal reaches a specific concentration and the cells can sense. This triggers genetically regulated changes that cause cells to bind tenaciously to the substrate and to each other

Chapter 5 Environmental Influences and Control of Microbial Growth 5.1 Environmental Limits on Microbial Growth