10 (10, 1024) N=2 10 (1, 2) (5, 32) 0 1 2 3 4 5 67 8 9 10 11 Time (hours) (a) A. Use the graph above to describe the bacterial growth and scale seen. B. Describe the growth curve with a stationary phase that begins at hour 14 and a death phase 10 hours later. C. What are two means of measurement you would use to verify the prediction OLmade inart B
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- 8. A. Use Excel (or another graphing program) to draw the growth curve, In (X/X.) vs time, for bacteria grown in a 20 L suspension cell culture, given the following data: - initial concentration: 0.120 gdw cells/L Also report: - lag time: 1.5 hours - mass doubling time during exponential growth: 250 minutes - duration of exponential growth phase: 1 day (24 hours) - negligible time in the deceleration phase - 13 hours in the endogenous metabolism phase with no change in cell concentration - cell death rate with k = 0.0178 min -¹. B. What is the specific growth rate, µ? C. What is the maximum concentration of cells in the reactor? (gdw cells/L) and when does this occur? D. Other than time zero or the end of lag phase, at what time is the concentration of living cells in the reactor equal to the initial concentration of 0.120 gdw/L?use these OD numbers to plot growth curve for E. coli K12- and estimate generation time for this culture. Table 1. Absorbance (O.D.600) measured for growing culture of E. coli K12 (time course) Incubation time O.D.600 Incubation time O.D.600 O min 3 hrs 3 hrs 30 min 4 hrs 4 hrs 30 min 5 hrs 5 hrs 30 min 0.11 1.75 30 min 0.20 1.94 1 hr 1 hr 30 min 2 hrs 2 hrs 30 min 0.27 2.24 0.51 2.48 2.31 2.19 0.65 1.30A urine sample is diluted 1:1,000. If 0.1 ml were put into a Petri dish with melted agar, how many CFU/ml are in the urine if 158 colonies grow?
- You are cultivating Escherichia coli in a chemostat culture. The maximum specific growth rate (µmax) of E. coli that you can reach is known as 1.0 h-1 at your culture conditions. Describe what you would observe for each condition if you have the following settings: F (flow rate of the fresh medium into the bioreactor vessel) (L/h) V (Volume of the liquid culture in the bioreactor vessel) (L) a) 1 1 b) 5 4 c) 1 2(b) A Food material containing Bacillus stearothermophilus PS1518 as an indicator organism ts subjected to heat sterilization at 121 C. Calculate the time required to reduce the organism to one tenth of the original number. Fo value for the organism is 4 minutes and the decimal reduction time , D, at 116°C is 40 minutes. Assume operation is at constant temperature of 121°C. HINT Fo = -To 10 dt Where Fo = equivalent exposure time at 121°C of the actual exposure time at a variable temperature To = the reference temperature =121 C, z=10 = number ofC necessary for10fold increase in FA culture of S. cerevisea has an overnight OD of 2.3 (1.0 OD is approx 1.0x107 cells/ml) You will be plating 100µl onto agar and want the final count of colonies on the plate to be around 300 colonies. How much of the 2.3 OD culture must you use to get a 500µl subdilution (with sterile water), so that you have diluted enough to get approx 300 colonies per 100ul
- The data below were obtained for the growth of a pure culture of Escherichia coli in nutrient broth at a temperature of 37°C. Determine (1) the specific growth rate and (2) generation time of E. coli and the duration of the (3) lag and (4) log (or exponential) phases. (ln 2 = 0.693) Time (h) 0 1 2 3 4 8 16 32 Bacterial No./mL 104.1 103.9 104.4 105.5 106.5 107.7 108.0 107.6a. Let’s say for example that a milk sample has 10,000 bacteria per milliliters. If 1 mL of this sample were plated out, these would theoretically be 10,000 colonies in the Petri plate. Discuss and explain the serial dilutions of this example. b. The disk diffusion method was used to evaluate 3 disinfectants. The results were as follows: Solution Zone of inhibition X 0 mm Y 5 mm Z 10 mm How would you interpret? Provide inference.E. coli has a doubling time of 0.345 hours at 37 0C. Starting with 1 milligram of cells, assuming a lag phase of 30 minutes and a stationary phase of 30 minutes, what number of cells would you expect at the end of 13h, assuming that besides the lag and stationary phases the culture is continuously in exponential phase. Assume that ln (2) = 0.69 and that the mass of a single cell is 10^(-12) g.
- A bacterial culture is initially composed of 100 cells. After 1 hour the number of bacteria is 1.5 times the initial population. a. If the rate of growth is proportional to the number of bacteria present, determine the time necessary for the number of bacteria to triple. b. What is the time required for a culture with 1x106 of the same bacteria to triple? Explain your results. c. Under what conditions would the answers obtained in (b) be invalid?A pure bacterial culture of unknown concentration was diluted to determine the concentration of viable bacteria in the original culture. Serial dilutions were performed as 1. diagrammed below. Each dilution tube contained 400 ul of diluent and 100 ul was transferred into each tube. TSA plates were inoculated with 100 µul from the last three dilution tubes. a. What is the dilution between each tube shown in the diagram below? Express your answer as a ratio. b. What is the total dilution of tube number 5? Express your answer as a ratio. c. What is the concentration of viable bacteria in the original culture? Express your answer using scientific notation and the units CFU/ml. d. What is the concentration of viable bacteria in tube number 2? Express your answer using the units CFU/ml. e. If you inoculated a TSA plate with 250 µl from tube number 5, how many colonies would you expect to see after the plate was incubated? 1 2 3 5 423 80 13 Number of coloniesSerial dilution = previous dilution x dilution 11 mL [2] 3 mL Colony count: 100 0.1 mL sample [1] 9.9 mL 5 mL 0.1 ml. Cokony count Dilution: 10-1 10-3 [3] 150 Dilution factor (DF): [4] [5] [6] Compute for the CFU/mL of the sample. Show your solution and express your final answer to 2 significant figures.