Chapter 6: Microbial Growth How do bacteria grow? Not in size Increase in population size One cell divides into 2 new cells – binary fission Binary fission Binary fission: Attachment of chromosome to p.m.; replication of DNA; new p.m. and cell wall laid down between the 2 chromosomes This is the way that each new daughter cell gets one chromosome Number of generations Number of cells Log of # of cells 10X = # 0 1 0 1 2 0.3 2 4 0.6 3 8 0.9 4 16 1.2 5 32 1.5 6 64 1.8 7 128 2.1 8 256 2.4 9 512 2.7 10 1024 3.0 11 2048 3.3 12 4096 3.6 How can we describe growth? 2n = no. of cells in n generation Generation time Nt = N0 x 2n Growth problems If Staphylococcus aureus has a doubling time (generation time) of 30 minutes and 5 hours have passed, how many generations have been produced? a. How many 30 minute time chunks are in 5 hours? = 10 ANSWER: 10 generations Growth problems If Staphylococcus aureus has a generation time of 30 minutes and 5 hours have passed, how many bacteria will be present at the end of the time period? a. b. c. We have already determined that 10 generations will occur. 2n = # cells at n generation 210 = # cells at the 10th generation ANSWER = 1024 cells Growth problems If Staphylococcus aureus has a generation time of 30 minutes and 5 hours have passed, how many bacteria will be present at the end of the time period if we start with 3,000 cells? Nt = N0 x 2n Nt = 3,000 x 210 = 3,072,000 cells at the end of 5 hours Growth curve: lag, log, stationary, and death phases What occurs in each? How can population size be counted? (Advantages and disadvantages of each method) 1. Direct methods A. Microscopic count with hemacytometer/ Petroff Hauser counting chamber Petroff Hauser counting chamber B. Plate counts – dilution series and plates C. Filtration D. Coulter counter/flow cytometer/Fluorescence activated cell sorter (FACS) 2. Indirect methods A. Dry weight B. Metabolic activity C. Turbidity Turbidity Growth requirements of microbes A. Temperature: Thermophiles Mesophiles Psychrophiles B. pH: acidophiles C. Osmotic pressure (# of solutes in solution) Halophiles D. Oxygen: Types: Obligate aerobe Facultative anaerobe Obligate anaerobe Aerotolerant anaerobe Microaerophilic Enzymes needed to survive in presence of oxygen: Catalase Peroxidase Super oxide dismutase (SOD) E. Nutrients C, N, P, S elements needed Mg, Fe, etc. trace elements needed Media: Defined or complex Selective vs. differential Special Mannitol salt agar – selective medium Blood agar – differential medium The End