Bacterial Growth and Reproduction

Learning objectives
Define bacterial growth including binary fission Compare phases of microbial growth Describe various direct and indirect methods of measuring bacterial growth

Microbial growth

defined not in terms of cell size but as the increase in the number of cells, which occurs by cell division.

BACTERIAL CELL DIVISION

Budding- form a small initial outgrowth (a bud) that enlarges until its size approaches that of the parent cell, and then it separates Binary Fission -a type of Asexual reproduction in single celled organisms -Each cell divides into 2 equal sized offspring

Binary fission
1. The cell elongates and chromosomal DNA is replicated. 2. The cell wall and cell membrane pinch inward and begin to divide. 3. The pinched parts of the cell wall meet, forming a cross wall completely around the divided DNA. 4. The cells separate into two individual cells.

Generation Time
amount of time needed for a cell to divide Varies among organisms and depends upon the environment they are in and the temperature of their environment

Phases of Growth
Lag Log (logarithmic) Stationary Death

Lag Phase
Period of adapting to nutrients Can last for 1 hour or several days little or no cell division DNA and enzyme synthesis

Log/exponential phase
cells begin to divide and enter a period of growth or logarithmic increase. cells are most active metabolically sensitive to adverse conditions

Stationary phase
Phase of equilibrium growth rate slows, the number of dead microorganisms equals the number of new microorganisms, and the population stabilizes

Death Phase
number of dead cells exceeds the number of new cells Continues until the population is diminished or dies out entirely.

Bacterial Growth Curve

Direct Measurement of Microbial Growth

Plate counts - advantage: it measures the number of viable cells -disadvantage: that it takes some time, usually 24 hours or more, for visible colonies to form - reported as colony-forming units (CFU)

Serial dilution

Pour plate and spread plate method

Counting colonies.

Filtration

amount of bacteria is very small 100 milliliters of water are passed through a thin membrane( pores are too small for the bacteria to pass through). The bacteria that are retained from the filter are placed on a Petri dish containing a pad soaked with liquid nutrient

Most Probable Number (MPN) Method

statistical estimating technique based on the fact that the greater the number of bacteria in a sample, the more dilution is needed to reduce the density to the point at which no bacteria are left to grow in the tubes in a dilution series.

Direct Microscopic Counts

known volume of medium is introduced into a specially calibrated, etched glass slide called a Petroff-Hausser counting chamber/ hemocytometer

Estimating Bacterial Numbers by Indirect Methods

Turbidity -practical way of monitoring bacterial growth - measured by spectrophotometer

Metabolic activity
assumed that metabolic waste products, CO2 (carbon dioxide) and acid direct proportion to the number of bacteria present.

Dry Weight
For filamentous organisms, such as molds they must be separated from the medium by some physical means such as filtration or centrifugation. The cells are then dried, and the resulting mass is weighed.