BACTERIAL PHYSIOLOGY AND GROWTH MICROBIAL GROWTH: Microbial growth is the increase in number of cells, not cell size.

THE REQUIREMENTS FOR GROWTH: PHYSICAL REQUIREMENTS 1. TEMPERATURE 3 Major Classification: Psychrophiles (cold-loving microbes). Mesophiles (moderate temperature-loving microbes); pathogenic bacteria. Thermophiles (heat-loving microbes).

Minimum growth temperature. Optimum growth temperature. Maximum growth temperature.

Psychotrophy Grow between 0C and 20-30C. Cause food spoilage.

2. PH Most bacteria grow between pH 6.5 and 7.5. Molds and yeasts grow between pH 5 and 6. Classification: Acidophiles grow in acidic environment. Neutrophiles grow in neutral environment. Alkalinophiles grow in alkaline environment.

3. OSMOTIC PRESSURE Hypertonic environments, increase salt or sugar, cause plasmolysis. Extreme or obligate halophiles require high osmotic pressure. Facultative halophiles tolerate high osmotic pressure.

THE REQUIREMENTS FOR GROWTH: CHEMICAL REQUIREMENTS 1. ENERGY SOURCES Phototrophs use light as source of energy. Chemotrophs use chemicals as source of energy.

2. CARBONS Structural organic molecules, energy source (Chemotrophs). Heterotrophs use organic carbon sources. Autotrophs use inorganic carbon source such as CO2. Inorganic elements required in small amounts. Usually as enzyme cofactors. 1. Nitrogen In amino acids and proteins. Most bacteria decompose proteins. Some bacteria use NH4+ or NO3. 2. Sulfur 4. Oxygen (O2) A. Facultative anaerobes prefer to grow in the presence of oxygen, but are capable of growing well in the absence of oxygen. B. Obligate aerobes require oxygen in order to grow. This limits the body sites where these organisms can grow. C. Obligate anaerobes - oxygen is extremely toxic. D. Aerotolerant organisms do not utilize oxygen for metabolism but can tolerate the presence of some level of oxygen in the environment. E. Microaerophilic bacteria differ from aerotolerant in that these organisms may utilize oxygen when present at a reduced level; normal levels of oxygen are inhibitory. In amino acids, thiamine and biotin. Most bacteria decompose proteins. Some bacteria use SO42 or H2S. In DNA, RNA, ATP, and membranes. PO43 is a source of phosphorus.

3. TRACE ELEMENTS

A few bacteria use N2 in nitrogen fixation.

3. Phosphorus

5. Carbon Dioxide Capnophilic microbes grow best in an atmosphere containing increased CO2 (5-10%).

4. ORGANIC GROWTH FACTORS Organic compounds obtained from the environment. Vitamins, amino acids, purines, and pyrimidines.

REPRODUCTION IN PROKARYOTES Binary fission. Budding. Conidiospores (actinomycetes). Fragmentation of filaments.

BINARY FISSION Formula:

Generation time = the time for a cell to divide or double. If 1 cell has 20 minutes generation time, how many cells are present after 1 hour? Number of generations (N) = time/ generation time N = 60 minutes/ 20 minutes N=3

Number of bacterial cells

= 2N (number of generations) = 23 = 8 bacterial cells.

BACTERIAL GROWTH CURVE A. Lag phase Initial period of adjustment to a new environment. Cells are actively metabolizing in preparation for cell division. It is characterized by NO increase in cell number.

B. Log phase or Exponential phase It is during this phase that the organism is most active metabolically.The bacteria undergo a period of exponential growth. The period, in which the cells grow most rapidly, doubling at fairly constant rate. It is characterized by INCREASE in cell number.

C. Stationary phase The rate of cell death is equal to the rate of replication. Metabolism and cell division slows down due to depletion of nutrients and accumulation of waste products.

D. Phase of decline or death phase The rate of cell death will exceed the rate of replication. The cells lose the ability to divide and the population of viable cells declines.

MEASURING MICROBIAL GROWTH

PLATE COUNT 1. Plate counts: Perform serial dilutions of a sample. 2. Inoculate Petri plates from serial dilutions. 3. After incubation, count colonies on plates that have 25-250 colonies (CFUs). *A standard plate count reflects the number of viable microbes and assumes that each bacterium grows in a single colony (Reported as CFU). A plate count may be done by either pour plate or spread plate method.

FILTRATION

MPN Multiple tube MPN test. Count positive tubes and compare to statistical MPN table.

DIRECT MICROSCOPIC COUNT

TURBIDITY