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Question 88 - In the calculation of the growth of bacteria, colony forming unit (CFU) in serial dilution is used. In a laboratory, viable count assay is used to estimate CFU. Formula applied is CFU / mL = (number of colonies x dilution) / (amount plated, in unit mL). Acceptable plate count is either between 20 and 200 or between 30 and 300 according to 2 different references. A wastewater sample of 200 ml is added to and mixed with 1.8 L of sterile water. Another 200 ml of the mixture is added to and mixed with 1.8 L of sterile water. (a) Calculate the dilution of first mixture and the dilution of the second mixture. (b) 100 microlitres of wastewater samples from the first mixture and the second mixture are placed separately on 2 different alga plates. The first plate has 250 colonies and the second plate has 23 colonies. Calculate the average CFU / mL.

Question 90 - In the calculation of the growth of bacteria, absorbance, A in spectrophotometry is used. According to Beer-Lambert Law, A = e x l x c where A is the absorbance of the solution (no unit), l is the distance of light travels through the solution (in cm), e is the molar absorptivity or the molar extinction coefficient [ in L / (mol.cm) ]. For a particular solute and fixed path length : As / Ao = Cs / Co where Ao is the observed signal for a known concentration Co, and As is the observed signal for a sample concentration Cs. (a) For a cell concentration of 560 cells / mL, a spectrophotometre gives an absorbance reading of 1.0. A mixture of concentration 3600000 cells / mL can be diluted in several operations, with each operation having a dilution of 1:20. How many dilutions should be made so that the concentration of this mixture can be calculated within a range of A = 0.0 to 1.0. (b) In another experiment, a sample tube of 1 cm in width is used. Let A = 0.06 and e = 0.0012 ml / (cell.cm). Find the cell concentration of the sample.

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MICROBIOLOGICAL ENGINEERING - QUESTION 28.1 : In the calculation of the growth of bacteria, colony forming unit (CFU) in serial dilution is used. In a laboratory, viable count assay is used to estimate CFU. Formula applied is CFU / mL = (number of colonies x dilution) / (amount plated, in unit mL). Acceptable plate count is either between 20 and 200 or between 30 and 300 according to 2 different references. A wastewater sample of 200 ml is added to and mixed with 1.8 L of sterile water. Another 200 ml of the mixture is added to and mixed with 1.8 L of sterile water. (a) Calculate the dilution of first mixture and the dilution of the second mixture. (b) 100 microlitres of wastewater samples from the first mixture and the second mixture are placed separately on 2 different alga plates. The first plate has 250 colonies and the second plate has 23 colonies. Calculate the average CFU / mL.

MICROBIOLOGICAL ENGINEERING - QUESTION 28.3 : In the calculation of the growth of bacteria, absorbance, A in spectrophotometry is used. According to Beer-Lambert Law, A = e x l x c where A is the absorbance of the solution (no unit), l is the distance of light travels through the solution (in cm), e is the molar absorptivity or the molar extinction coefficient [ in L / (mol.cm) ]. For a particular solute and fixed path length : As / Ao = Cs / Co where Ao is the observed signal for a known concentration Co, and As is the observed signal for a sample concentration Cs. (a) For a cell concentration of 560 cells / mL, a spectrophotometre gives an absorbance reading of 1.0. A mixture of concentration 3600000 cells / mL can be diluted in several operations, with each operation having a dilution of 1:20. How many dilutions should be made so that the concentration of this mixture can be calculated within a range of A = 0.0 to 1.0. (b) In another experiment, a sample tube of 1 cm in width is used. Let A = 0.06 and e = 0.0012 ml / (cell.cm). Find the cell concentration of the sample.

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