Viability of Escherichia coli ATCC 8739 in Nutrient Broth, Luria-Bertani Broth and Brain Heart Infusion over 11 Weeks
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Samuel Xin Zher Low, Zhen Qin Aw, Bryan Zong Lin Loo, Kun Cheng Lee, Jack Si Hao Oon, Chin How Lee, Maurice Han Tong Ling
2013 Volume 5, Issue 1, p576-81
Abstract
Escherichia coli is a widely studied prokaryotic system. A recent study had demonstrated that reduced growth of E. coli after extended culture in Luria-Bertani broth is a result of depletion of fermentable sugars but able to sustain extended cell culture due to the presence of amino acids, which can be utilized as a carbon source. However, this had not been demonstrated in other media. The study aimed to determine the growth and viability of E. coli ATCC 8739 in 3 different media, Nutrient Broth (NB), Brain Heart Infusion (BHI) and Luria-Bertani Broth (LB) over 11 weeks.
Growth of E. coli ATCC 8739 was determined by optical density. Viability was determined by serial dilution/spread-plate enumeration. After 11 weeks, the media were exhausted by repeated culture. Glucose was added to the exhausted media to determine whether glucose is the growth-limiting factor.
Our results showed that cell density in all 3 media increased to about 1 × 10(9) cells/ml by the end of week 1, from the inoculation density of 2.67 × 10(5) cells/ml, peaked at about 1 × 10(13) cells/ml at week 4, before declining to about 5 × 10(7) cells/ml at week 7. Cell density is highly correlated to genomic DNA content (r(2) = 0.93) but poorly correlated to optical density (r(2)< 0.2). Our results also showed that the spent media were able to support further growth after glucose-supplementation.
NB, LB and BHI are able to support extended periods of culture and glucose depletion is the likely reason for declining cell growth.
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