I. Microbial growth II. Environmental effects on microbial growth

I. Microbial growth A. Exponential growth and how to calculate it B. Growth curve of bacteria in culture C. Measuring cell growth D. Continuous culture

I. Microbial growth A. Exponential growth and how to calculate it

Exponential growth

Text Fig. 7.16

Generation time Generation time (g) (or Doubling time) -- the time required for a population of cells to ___________. Or: the time required for one cell to become ______. For this example, g = ___________

Estimating generation times ___ hr

Estimating generation times (cont.) ___ hr

Calculating generation times (g) = 2 0 = 2 1 = 2 2 = 2 3 = 2 4 = 2 5 = 2 6 = 2 7 = 2 8 = 2 10 = 2 9 = 2 11 = 2 12 Notice the simple “geometric” progression of the number 2 = 2 6

Calculating generation times (cont.) Because of this geometric progression, there is a direct, predictable relationship between the number of cells in a culture ________ and the number there ___________ a period of exponential growth: Nf =Nf =Nf =Nf = Where: N f is the ________ cell number N i is the_________ number of cells n is the _______________________

Calculating generation times (cont.) N f = (N i )2 n So, if we start with 20 Salmonella cells in a potato salad sitting on your warm kitchen counter, with a 30 minute generation time, how many cells will there be after 8 hours? To get the # of generations (n), divide 8 hours by 30 minutes to get 16: N f = (20)2 ___ N f = (20)2 ___ N f = (20)(_______) = 1,310,720 cells

Why haven’t bacteria, then, grown to fill the entire earth? Why aren’t we knee deep in them? Answer: eventually something _________ their __________. Such as? --lack of __________, build up of ___________, environmental conditions, predation, etc. A single bacterium with a generation time of 20 minutes would, growing exponentially for 48 hours, produce a population weighing approx times the weight of the earth! (And a single bacterium weighs approx. one-trillionth of a gram)

The Bacterial Growth Curve

Text Fig _____ phase -- period after inoculation of a culture before growth begins. Exponential phase -- period of exponential growth _______________ phase -- period in which growth ceases Death phase -- period in which cells die off

Growth curve animations

How do we measure growth? Total cells count (or “direct microscopic counts” or just “_________ ___________”) Total cells count (or “direct microscopic counts” or just “_________ ___________”) Viable counts (also called “___________ ____________” or “colony counts”) Viable counts (also called “___________ ____________” or “colony counts”) Measurements of cell mass and _____________ Measurements of cell mass and _____________

a.k.a “direct counts”, “total cells counts” Text Fig. 7.16

_________ plate method _______ plate method Sample pipetted onto agar surface Sample spread evenly over surface Incubation Typical results Sample pipetted into sterile plate Sterile medium added and mixed well with inoculum Incubation Typical results Viable count

Viable counts using ___________ ___________

Measurements of cell mass and turbidity 1. Measuring ________ __________ : centrifuge organisms and _________ them (usually dried). Cell mass is proportional to cell number 2. Measuring ________________ : measure cloudiness of a cell suspension by light scattering. Turbidity is proportional to cell number, also Turbidity is proportional to cell number, also

Measurement of turbidity using a ____________________

1 2 Typical growth curve data (obtained in Klett units) for two organisms growing at different growth rates

Cell number or dry weight and turbidity are ___________________ In other words, once you know one, you can figure out the other if you know their relationship

Chemostat -- a device for continuous culture

Effect of environmental factors on microorganisms 1) Important to distinguish between effects on _____________ (whether the cells survive) and effects on ____________ (including reproduction). 2) Important factors:  _____________  _________ availability  oxygen

Effect of temperature on growth rate Each microorganism has a minimum, maximum, and ___________ Each microorganism has a minimum, maximum, and ___________ Organisms can be classified by temperature Organisms can be classified by temperature Text Fig. 7.16

Effect of temperature on growth rate

Boulder Spring, a boiling spring in Yellowstone National Park

Growth of thermophilic cyanobacteria in Grand Prismatic Spring, Yellowstone

Photomicrograph of a bacterial _________________ which developed on a microscope slide immersed in a boiling spring

RESEARCHERS DISCOVER EVIDENCE OF MICROSCOPIC LIFE AT THE SOUTH POLE July 6, 2000 In a finding that may extend the known limits of life on Earth, researchers have discovered evidence that microbes may be able to survive the heavy doses of ultraviolet radiation and the extreme cold and darkness of the South Pole. The team's findings, published in Applied and Environmental Microbiology, the journal of the American Society for Microbiology, indicate that a population of active bacteria, some of which have DNA sequences that align closely with species in the genus Deinococcus, exists at the South Pole in the austral summer. A similar species lives elsewhere in Antarctica, but the discovery of microbes at the Pole may mean that the bacteria have become uniquely adapted to the extreme conditions there, including a scarcity of liquid water. A species in the genus Deinococcus was first discovered in cans of irradiated meat in the 1950's, and is able to withstand extreme dryness and large doses of radiation. It is possible that the related bacteria from the South Pole may also possess these characteristics. "While we expected to find some bacteria in the South Pole snow, we were surprised that they were metabolically active andsynthesizing DNA and protein at local ambient temperatures of -12to -17 Celsius (10.4 to 1.4 degrees Fahrenheit)," said Edward J. Carpenter, of the State University of New York at Stony Brook.

Cytoplasmic pH ________________ _______________ Effects of pH

Effects of salt (sodium ion concentration) on _________ ___________ of various microorganisms

Text Fig. 7.16

Toxic forms of oxygen

Enzymes acting on toxic oxygen species

Method for testing a microbial culture for the presence of ____________