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    MCB 309

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    MCB 309

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    MCB 309

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    Emmanuel ogenyi

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    WA pe _ watnon é o | COURSE TOPICS. ee edi Jeroblal Growth | MICROBIAL PHYSIOLOGY, + Effoct of physical and chornical factors un | AND METABOLISM imlcrobial growth + Dynamies of growth and 7 + Growth measurements DR. |. W NYINOH Microbial Enzymos COURSE LECTURER + Classification, Proportios, Mode of action + Mode of Production and regulation i LEARNING OUTCOMES | + Upon successful completion ofthis topic, + Predict the growth behavior of |__ you should be able to microbes based on their growth \ | + Describe how very high (or low) ‘conditions, e.g., temperature, f a |. _ttuparatures; oH sal Conc Toe oF available nutrient, aeration level, etc ther factors inhibit microbial-grow . |v ae + Explain two strategies that are used |, + Describe how oxygen affects the growth of ff numnan thod reciaratioriio \ aerobes, obligate anaerobes, and. facultative anaerobes minimize microbial growth during storage Lecture 1: Microbial Growth Microbial Growth + While growth common refers to increase in size + but with microorganisms particularly Introduction + Living organisms (plants, animals and s microbes) grow and reproduce - bacteria, + The growth indicates that an + Microblal growth refers to the organism is in active metabolism growth of a population (or an increase + In plants & animals, we can see the in the number of cells) increase in length, height or size * notto an increase in the size of the - l individual cell ? * The change in population in-bacteria chiefly involves binaty fission * Therefore, a single bacteri Continuously divides © =" * 1 cell divides and providing 2 cells and 2 cells divide Providing 4 cells | and so on + Therefore, the Population increases by geometric Progression ion Control of growths important to: * prevent thé transmission of diseases:and infection (in infection diseases:e.g, TB) + stop decomposition and industry) * and prevent unwanted microbial contamination (in'infection ‘control in hospitals, cosmetics and pharmaceutical) Microorganisms are controlled by means of Physical agents and chemical agents = Cells will divide untilthey are subjected to stress: +, nutrient depletion 7 oF environmental stress (e.g. change in temperature) Environmental Factors affecting growth + Broadly sub-divided into physteal and chemical requirements Physical requirements: + Temperature > pH + Moisture + Osinotie Pressure + Others: radiation, atmospheric pressure + Chemical requirements encompass nutritional requirements | Temperature Température is an important factor that determines: + the rate of growth * multiplication + survival, and = + death of all living organisms 4. -High temperatures damage microbes by: + denaturing enzymes- Microbial membrane are disrupted by temperature extremes + transport carriers, and + other proteins 2. Atvery low temperatures membranes also solidity and enzymes also do not function properi 3/22/2021 eS i ‘ Classification of microbes based A. Psychrophiles:"Cold-loving” on temperature ate : + Microbes are loosely classified into several ss Esychiol fils iE SEOpUIIC: Beas groups based on their preferred temperature ranges + The two major groups are: 4. Psychrophiles and 2. Psychrotroph (Psychrotelerant psychrophiles) rganisms that are capable}. — reproduction:in cold] + Temperature range: On. | . + They grow at refrigeration ter peratures [ = | Most are bacteria-or-archae, but also 5. | fungi and species of yeast Psychrotrophs: | + Most members belong'to the genus + Psychrotrophs are cold-tolerant bacteria | Psychyobacter spp:, Aeromonas, ‘or archaea that have'the ability to grow at |. Flavobacterium, Serratia, Arthrobacter low temperatures, | and Methanogenium, Halomonas, + but have optimal and maximal growth | Sphingomonas temperatures above 15 and 20°C, respectively : +Optimum growth at 20 to 30°C + Responsible for most low temperature food spollage e.g. milk spoilage | + Pseudomonas fluorescens L | + Gram+ve: Bacilus, Clostridium, | Micrococcus | . |B. Mesophiles: “Middle loving” C. Thermophiles: “Heat loving” | * + Most bacteria belong to this category + Derived from Greek word thermotita + includes most human pathogens meaning heat and philia meaning love | and common spoilage organisms + Thermophiles are a group of heat + Best growth between 25 to 40°C loving microbes thriving at high = Optiqum temperature:cpmmanie temperature usually more than 45°C 37°C + Optimum growth between 50 to 60°C + Marly have adapted to live in the + Some thermophiles form extremely bodies of animals.£.g. Escherichia heat resistant endospores coli, Streptococcus pneumoniae ; + They are inhabitants of various ecological niches like deep sea hydrothermal vents, terresttial Hot springs, and other extreme * geographicaligeological sites. including volcanic sites, tectonically active faults as well z matters such-as the com and deep organic | «Extreme Thermophiles - : “Thermophiles ‘contd.: + Eg. green-and purple-sulfurbacteria ee shromatium and Chlorofiexus, “Thermus aquaticus, Geogerma:barossii, ate 5 : (Hyperthermophiles): = Optimum growth at 80°C-or higher. Eg. Archaebacteria. . e in volcanic and ocean vents. Tigao pH + pHrefers to negative logarithm of hydrogen ion concentration Microbial growth is strongly affected by the pH of thie medium 1. “Drastic variations in cytoplasmic PH disrupt the plasma membrane 2. or inhibit the activity of enzymes and 3._membrane. transport protein! + Most bacteria prefer neutral pH (6 57 5) * Molds and yeast orow in wider pH:range: but orefer pH between 5 and 6 + Acidity * Acidic produets of bacterial metabolism Interfere with growth. Buffers can be used to stabilize pH * Acidity inhibits most microbial growth and 's used frequently for food preservation (€.9: picking of oives) * Alkalinity intibts mierobia growth, but {__not commonly used for food preservation Based on pH bacteria can ‘classified as: 1. Acidophiles (acid-loving) ~ grow bestata pH of 1 to 5.4; Eg. Lacfobacilllus (ferments milk) + preservative for pickles, sauerkraut, cheeses + Many bacteria and viruses survive low pH of stomach to infect intestines + Helicobacter pylori lives in stomach under mucus layer 2. Neutrophiles — exist from pH of 6.4 to 8.5; most pathogens that cause human disease are in this category + E.g &, coli, Pseudomonas aerunginosa 3. Alkaliphiles (base loving) — Grow at alkaline or high pH (7 to. 12 or higher), + Eg. Vibrio cholerae (causes cholera) and Alkaligenes faecalis optimal-pH 9. + Soil bacterium Agrobacterium grows at pH12 2 . + All microorganisms req and reproduce. growth varies be ‘The amount of water that is avallable in food | -. igtexpressed in terms of water activity (aw) \ Water activity vs Water Content | |. Water content describes how much | _ water is physically presentin a \ product whereas i water activity describes how much if that waters ‘free’ ea \ + Only ‘free’ water can participate in chemical reactions, migrate throughout the product, or be used by microorganisms! + ‘The water activity is expressed in’a value “ranging from 0 to + where the aw of pure water is 1.0 + Each microorganism has a maximum, ‘optimum, anc inimum aw for growth and survival = + Generally bacteria dominate in'foods with high aw (minimum approximately ogvaw ‘ 7+ while yeasts and moulds, which: regula less moisture,dominate in.low aw foods (minimum 0,70 aw) és + The water activity of fluid milk is approximately. 0.98 aw | + Pathogenic bacteria do not grow well ol or produce toxin below 0.85 and most. require 0.92 or above = * + Freezing, drying, or salting are ways to reduce available water to bacteria, and slow down their growth + This is why they grow on foods with high moisture content such as chicken ‘The aw at which no microorganisms grow (<0,6) ‘At an aw-Value'smaller than 0.6, virtually ‘no microorganism will be able to grow. ‘There just isn’t enough water. Crackers, driod pasta, dried spices, noodles, and flour are examples of foods that can have such a low water activity Because of their low water activity, spoilage by microorganisms isn't a concern as long as they are stored properly How to lower the water activity to prevent microbial growth + Before the availability of fridges or widespread cooling, which greatly slows down growth of microorganisms, people needed to. find other ways to preserve foods * This was mostly done by lowering the water activity of foods t&:for instance: greatly their water activity and inhibits “the growth: of harmful. mictooryanisms. helping:to extend its shelflife «Salt. binds" the: water, making it less accessible: » Another commonly: used-method is to + Sugar also ‘binds’ water ana makesitless. available + Eg making jam out of fruits + Alternatively, you can dry. foods, By simply decreasing the overall amount of water in the food, you:lower the water activity of thatfood. + Drying fruits (dates, aya), vegetables, and meats (dry fish, bush meat) are all Very effective ways to lower the water activity and'extend the: shelf life of the food by preventing growth of Osthotic pressure versus Tonicity + Cells are 80 to 90% water + Diffusion of water actoss a membrane generates a pressure called osmotic pressure * Osmotic. pressure is the pressure of a solution against a semipermeable membrane to prevent water from flowing inward across the membrane + Tonicity is the measure of this pressure. : + The tonicity of solutions with respect to each other may’be expressed as hypertonic, isotonic or hypotonic + The effect of different external solute ‘concentrations on’ted blood cells serves as a good.example fora hypertonic, isotonic and hypotonic solution. ‘A. Hypertonic solutions or Hypertonicity + High osmotic pressure removes water from cell, causing shrinkage of call membrane (plasmolysis) + When the osmotic pressure of the solution outside the blood cells higher than the osmotic pressure inside the red blood cells; the solution is hypertonic 3/22/2021 ‘The water inside the blood cells exits the cells in an attempt to equalize the osmotic pressure, causing the cells to shrink + When the osmotic pressure outside the red blood cells is the same as the pressure inside the cells, the solution is isotonic with respect to the cytoplasm = This is the usual condition of ted { blood cells in plasma >. Hypote feaotutionsor 3 Hyot + When the solut blood cells has a lows pressure than the cytoplasm of the ted blood cells, the solution is hypotonic with respect to the cells + The Cells take in water in-an attempt to equalize the osmotic pressure, causing them to swell and potentially burst . + In most cases cell wall prevents ‘excessive entry of water + Microbe may lyse or burst if cell wall is weak Effects of osmosis on bacteria cells 4, Osmotolerant Waco cms engin ot ies Nec, Osmotolerance-of halotolerance |. « Osmotolerant are those ticroorganisms which can survive | and.grow at high:osmotic ncentrations (relatively high salt or ’ “con ‘sugar Concentration) that reduces aw “Examples: Aeromonas spp., Staphiylacocous spp. eto. ‘ [exons or Obligate Halophiles: Require very high-salt concentrations “(20:0 30%). : z + Bacteria in Dead Sea, brine vats Jarge:salt _+ Require moderat E jegan water contains. concentrations. + Eig. Most bacteria'in-oceans a c a . + Examplés: Halobacterium halobium | . bs sto Facultative Halophile: Other physical factors * Do not require high salt » Radiation - UV rays and gamma rays concentrations for grawth, but tolerate ‘can cause mutations in DNA and Se even kill microorganisms 2% salt or more se + Some bacteria have enzyme systems that can repair some mutations. 3/22/2021 Chemical factors affecting microbial growth * The chemicals and elements of thie V2 environment that are utilized for bacterial - growth are referred to as nultients.or nutritional requirements + Inthe laboratory, bacteria are grown'in culture media which are désigned'to provide all the essential nutrients in solution for bacterial growth inthe form 1s; small. lecules, and macromolecules ¢ serve eithera structural or. A Mute: Hc therats and eltnents of tee Our] Nien thet ae ube droit. _functional:rolein the cell 1. Oxygen: + Organisms that use molecular oxygen: (02), produce more'energy from nutrients than anaerobes + Can classify microorganism based on their oxygen requirements: : A ‘Obligate \erobe: + Require oxygen todive. {ot qos +-oni{ aerobic growth) + oxygen Lequired:-growth-eeeurs-with high-soacentration.of oxygen | + Example: Pseudomonas, common nosocomial pathogen > B. Facultative Anaerobes: + Can use oxygen, but can grow in its absence (both aerobic and anaerobic. growth) + can shift their metabolism (anaerobic if ‘oxygen is absent or aerobic if oxygen is present) + Have complex set of enzymess + Examples: E. col, Staphylococcus, yeasts, and many intestinal bacteria C. Obligate Anaerobes: + Cannot use oxygen andiare’harmed by the presence of toxic forms of oxygen + only anaerobic:growth + i.e. growth ceases in the presence of oxygen; growth occurs only when there is no oxygen + Examples: Clostridium bacteria that cause tetanus and botulism, Rouel VOZuWemsfs' ave Bred for bacterial 7 D. Aerotolerant Anaerobes * only anaerobic growth, but cont but contin in pres ygen he 0 Geresence of oxygen, xygen has no * Le. Can't use oxygen: but tol tus : lerate Presence. Can break down: ce 2 3 3/22 concentrations reset y 2 ie mS ot Oxygen : * Sensitive to toxic forms'of oxygen. mple: Lactobacillus carries out e ‘Campylobacter’ »xyGEN presence. t ‘ane © owls Stasbes - snkroasoaes 2. Carbon * Carbon: Makes up 80% of dry weight of cell + Structural backbone of all organic ‘Sompounds Carbon sources « + C02. . = autotroph | + organic, = heterotroph + Chemoheterotrophs : Derive both’carvon ‘and energy from organic compounds: lipids, proteins, and carbo + Chemoautotrophs ‘and Photoautotrophs : ‘Obtain carbon from carbon dioxide. Chemoorganic autotroph Derives energy from organic compounds and carbon source from inorganic compounds Arelated ancient group. Lithoautotroph Neither sunlight nor organics used, rather it relies totally on fs inorganics 3/22/2021 * Nutritional Categories + Saprobe — lives on organic matter of dead organisms + Parasite — lives on organic matter of living host = pathogens 2: Nitrogen; Sulfur,and Phosphorus: + -ALNitrogési- Makes up 14% of dry cellweight. Usedto form amino acids, ONA: and RNA ater © realy om’ ee eae “+ Important nitrogen fixing bacteria, ive free in soll ne ee wi gu = ‘beans, alata, See ee emcees eles te i 2. Nitrogen; Sulfur, and Phosphorus: + B Sulfur: Used to form proteins and some vitamins (thiamin and biotin) + Sources of sulfur: Protein: Most bacteria Hydrogen sulfide Sulfates: Salts that dissociate to give so C:Phosphorus: Used to form DNA, RNA, ATP, and phospholipids: ‘Sources: Mainlyinorganic phosphate salts and buffers r | 3. Other Elements : + Potdssium, magnesium, and calcium are often required as enzyme cofactors. + Calcium is required for. cell wall synthesis in Gram positive bacteria “q.Trace Elements: : ‘ Many-are-used as enzyme cofactors. ‘Commonly found in tap water Iron Copper Molybdenum Zine rc Classification, Properties, Mode of action of Enzymes types of cells, tissues and other complex| organs. + For efficient functioning, our body releases| some chemicals to accelerate biological digestion, excretion and few ~ other| metabolic activities to. sustain a-healthy| fe > The human body is composed of different] ° processes such as in. respiration.) — * 5/31/2021 —] Enzyme Classification + According to. the International: Union of Biochemists (| U B), enzymes are divided! into six functional. classes and . are| classified based on the type of reaction in| which they catalyze. 4, Oxidoreductases: These »-catalyze| oxidation and reduction reactions, \ 5. | pyruvate dehydrogenase, catalysing the| | oxidation of pyruvate tg acety! coenzyme - TR 2.Transferases |. These: catalyze the, transfer.of a chemical group from one to-another compound. - An example is a transaminase, which) transfers ‘an amino” group. from one molecule to-another, dees .3. > Hydrolases: They’ catalyze _ the| hydrolysis of'2 bong For

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