Siti Norbaizura bt Md Zain Applied Science Faculty, UiTM Lecture 8 02/09/09

Bacterial Growth Definition

Vegetative growth

Cells actively growing and dividing

Microbial growth

Population growth as opposed to cell growth Measured by total number of cells Exponential due to binary fission reproduction The increase in number of cells, not cell size

Generation time (or doubling time)

The time for an individual or a population to double

 Binary

fission Budding

The bacterial chromosome is duplicated shortly before binary fission begins. The duplication usually occurs by the rolling circle mechanism taking place at the cell membrane. The duplicate chromosomes attach to the cell membrane, which grows and separates the chromosomes. At this point, the bacterium contains two chromosomes in regions known as nucleoids. The chromosomes separate, and the cell wall and cell membrane begin to form a septum. The septum becomes complete and the two new daughter cells separate.

 In

yeast, DNA duplication occurs and a new cell develops at the surface of the existing cells This small cells gathers cytoplasm and organelles (if a eukaryotic cell) and develops and breaks free from the parent cell The reproductive process is called budding

Binary Fission Generation Time (Doubling Time)

time required for a cell to divide most about 1 Hr. To 3 Hrs.
E. coli - 20 minutes Mycobacterium tuberculosis - 24 Hrs.

E. coli - generation time of 20 min. 20 generations (about 7 hrs.)

1 million cells

30 generations ( about 10 hrs.)

1 billion cells

72 generations ( about 24 hrs.)

1 x 1021
1,000,000,000,000,000,000,000 cells

If 100 cells growing for 5 hours produced 1,720,320 cells:

Direct methods Plate counts Filtration MPN Direct microscopic count Dry weight

Indirect methods Turbidity Metabolic activity Dry weight

Plate counts: Perform serial dilutions of a sample

Figure 6.15, step 1

Inoculate Petri plates from serial dilutions

Figure 6.16

After incubation, count colonies on plates that have 25-250 colonies (CFUs)

Filtration

Direct microscopic count

Turbidity

Bacterial Growth

Four phases of growth in a culture Lag phase Log phase (or exponential phase) Stationary phase Death phase (or decline phase)

 Will

encompass several hours During this time the organisms grow in size, accumulate organic matter and store large quantities of chemical energy such as ATP for biosynthesis

Microorganisms undergo rapid cell division and fulfill their generation time. The population doubles during each generation time The population increases in size at a logarithmic or exponential rate Rapid cell growth (exponential growth) population doubles every generation microbes are sensitive to adverse conditions

antibiotics anti-microbial agents

During the third phase of a populations history, the rate of cell division decreases and the older cells begin to die. During this phase, the number of living cells in the population remains constant. Death rate = rate of reproduction cells begin to encounter environmental stress lack of nutrients lack of water not enough space metabolic wastes oxygen pH Endospores would form now

During the fourth phase, the death/decline phase, the environment has become difficult for living and the rate of cell death exceeds the rate of cell division. The population declines in numbers and the population may completely die out if the environmental stresses are not reversed. Death rate > rate of reproduction Due to limiting factors in the environment

Generation time = Doubling time

Step 1: Get your growth curve (straight line with 3 points). Step 2: Pick any point above your time zero reading. Follow across to intersection of growth curve then down to time. Record this time. Step 3: Double the number you picked in step two and repeat step two. Step 4: subtract time 1 from time 2 = Doubling time.