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.
Answer 88 - (a) Dilution = (final volume of mixture) / (initial volume of mixture). First dilution = ( 0.2 + 1.8 ) L / (0.2 L) = 10. Second dilution = first dilution x ( 0.2 + 1.8 ) L / (0.2 L) = 10 x 10 = 100. (b) 100 microlitres = 100 microlitres x 1 mL / (1000 microlitres) = 0.1 mL. CFU / mL = (number of colonies x dilution) / (amount plated, in unit mL). In the first plate, CFU / mL = 250 x 10 / 0.1 = 25000. In the second plate, CFU / mL = 23 x 100 / 0.1 = 23000. Average CFU / mL = (first plated value + second plated value) / 2 = (25000 + 23000) / 2 = 24000. The answer is given by Kang Chuen Tat; PO Box 6263, Dandenong, Victoria VIC 3175, Australia; SMS +61405421706; chuentat@hotmail.com; http://kangchuentat.wordpress.com.
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A distillation column separates 10000 kg / hr of a mixture containing equal mass of benzene and toluene. The product D recovered from the condenser at the top of the column contains 95 % benzene, and the bottom W from the column contains 96 % toluene. The vapor V entering the condenser from the top of the column is 8000 kg / hr. A portion of the product from the condenser is returned to the column as reflux R, and the rest is withdrawn as the final product D. Assume that V, R, and D are identical in composition since V is condensed completely. Find the ratio of the amount refluxed R to the product withdrawn D. Hint : Solve the simultaneous equations as follow in order to find the answer (R / D) : 10000 = D + W; 10000 (0.5) = D (0.95) + W (0.04); 8000 = R + D.