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Exposure Effects of Aspirin on Growt...

Obanla, Temitayo Olumakinwa.

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Exposure Effects of Aspirin on Growth and Functionality of Lactobacillus rhamnosus and the Development of a Mutant Strain Using Chemical Mutagenesis.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Exposure Effects of Aspirin on Growth and Functionality of Lactobacillus rhamnosus and the Development of a Mutant Strain Using Chemical Mutagenesis./
Author:	Obanla, Temitayo Olumakinwa.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2017,
Description:	143 p.
Notes:	Source: Dissertation Abstracts International, Volume: 78-12(E), Section: B.
Contained By:	Dissertation Abstracts International78-12B(E).
Subject:	Food science. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=10264290
ISBN:	9780355055009

Exposure Effects of Aspirin on Growth and Functionality of Lactobacillus rhamnosus and the Development of a Mutant Strain Using Chemical Mutagenesis.
Obanla, Temitayo Olumakinwa.

Exposure Effects of Aspirin on Growth and Functionality of Lactobacillus rhamnosus and the Development of a Mutant Strain Using Chemical Mutagenesis. - Ann Arbor : ProQuest Dissertations & Theses, 2017 - 143 p.

Source: Dissertation Abstracts International, Volume: 78-12(E), Section: B.

Thesis (Ph.D.)--North Carolina Agricultural and Technical State University, 2017.

The objective of this study was to determine the effects of short and long term exposure to aspirin on the growth and functionality of Lactobacillus rhamnosus and to develop L. rhamnosus mutant that could improve functionality of L. rhamnosus on exposure to aspirin using chemical mutagenesis. One isolated colony of L. rhamnosus was activated in de Man, Rogosa and Sharpe (MRS) broth and incubated at 37°C until optical density of 0.9 was reached. The active culture (O.D. = 0.9) was then harvested and washed with 0.1% peptone water. The harvested cells were then exposed, short term (0-120min), to different concentrations of aspirin. Cell growth was assessed by measuring turbidity, and the population was determined by pour plate method. The minimal inhibitory concentrations (MICs) of aspirin were determined using the standard agar dilution method. The effects of aspirin on beta-gal activity were measured in miller units. The effects of long term exposure to aspirin on the growth of L. rhamnosus was determined by transferring 1mL of appropriate dilution (10-3) of active cells into 9mL of MRS broth containing approximately 6 mg/mL of aspirin. This mixture was vigorously mixed and incubated for 4h. The cells were again harvested, transferred into MRS broth (without aspirin), and incubated at 37°C until optical density of 0.9 was reached. The exposure protocol was repeated for five sequential transfers within a week and the exposed strain cell was then surface plated onto MRS agar containing an equal amount of aspirin. One isolated colony of this plate was then activated in MRS broth. This procedure was repeated sequentially for 12 consecutive weeks. A classical chemical mutagenesis protocol was used to evaluate the possibility of evolving resistance to aspirin and for increasing the production of beta-galatoctosidase. In order to carry out the mutagenesis, L. rhamnosus was exposed to ethyl methanesulfonate (EMS), a mutagen. Cells were screened to identify mutants that had high beta-galatoctosidase activity despite being exposed to aspirin. Cells were plated on MRS agar containing 5-bromo-4-chloro-3-indoly- beta-Dgalactopyranoside (X-gal). Colonies that displayed a blue color were selected for further analysis. Strains that had not been exposed to either aspirin or a chemical mutagen were used as controls for this study.

ISBN: 9780355055009Subjects--Topical Terms:

3173303
Food science.

Exposure Effects of Aspirin on Growth and Functionality of Lactobacillus rhamnosus and the Development of a Mutant Strain Using Chemical Mutagenesis.
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100 1 $a Obanla, Temitayo Olumakinwa. $3 3286247
245 1 0 $a Exposure Effects of Aspirin on Growth and Functionality of Lactobacillus rhamnosus and the Development of a Mutant Strain Using Chemical Mutagenesis.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2017
300 $a 143 p.
500 $a Source: Dissertation Abstracts International, Volume: 78-12(E), Section: B.
500 $a Adviser: SALAM A. Ibrahim.
502 $a Thesis (Ph.D.)--North Carolina Agricultural and Technical State University, 2017.
520 $a The objective of this study was to determine the effects of short and long term exposure to aspirin on the growth and functionality of Lactobacillus rhamnosus and to develop L. rhamnosus mutant that could improve functionality of L. rhamnosus on exposure to aspirin using chemical mutagenesis. One isolated colony of L. rhamnosus was activated in de Man, Rogosa and Sharpe (MRS) broth and incubated at 37°C until optical density of 0.9 was reached. The active culture (O.D. = 0.9) was then harvested and washed with 0.1% peptone water. The harvested cells were then exposed, short term (0-120min), to different concentrations of aspirin. Cell growth was assessed by measuring turbidity, and the population was determined by pour plate method. The minimal inhibitory concentrations (MICs) of aspirin were determined using the standard agar dilution method. The effects of aspirin on beta-gal activity were measured in miller units. The effects of long term exposure to aspirin on the growth of L. rhamnosus was determined by transferring 1mL of appropriate dilution (10-3) of active cells into 9mL of MRS broth containing approximately 6 mg/mL of aspirin. This mixture was vigorously mixed and incubated for 4h. The cells were again harvested, transferred into MRS broth (without aspirin), and incubated at 37°C until optical density of 0.9 was reached. The exposure protocol was repeated for five sequential transfers within a week and the exposed strain cell was then surface plated onto MRS agar containing an equal amount of aspirin. One isolated colony of this plate was then activated in MRS broth. This procedure was repeated sequentially for 12 consecutive weeks. A classical chemical mutagenesis protocol was used to evaluate the possibility of evolving resistance to aspirin and for increasing the production of beta-galatoctosidase. In order to carry out the mutagenesis, L. rhamnosus was exposed to ethyl methanesulfonate (EMS), a mutagen. Cells were screened to identify mutants that had high beta-galatoctosidase activity despite being exposed to aspirin. Cells were plated on MRS agar containing 5-bromo-4-chloro-3-indoly- beta-Dgalactopyranoside (X-gal). Colonies that displayed a blue color were selected for further analysis. Strains that had not been exposed to either aspirin or a chemical mutagen were used as controls for this study.
520 $a The result for short-term exposure showed significant dose-dependent reductions in the population of L. rhamnosus. The minimum inhibitory concentration (MIC) of aspirin on L. rhamnosus was determined to be 2mg/mL with an inhibitory zone diameter of 10 +/- 0.33mm. The average beta-gal activity of the control L. rhamnosus culture was 150 +/- 2.5Gal U. The beta-gal activity as the concentration of aspirin increased. L. rhamnosus could survive long term exposure to a sub inhibitory concentration of aspirin. The average amount beta-gal activity of unexposed cells was 153 +/- 2.5 Gal U.; however, the beta-gal activity was completely inhibited in exposed cells throughout the exposure period. There was approximately 54% more protein expressed as a result of long term exposure compared to short-term exposure (4h). In addition, the SDS-PAGE analysis confirmed this result. Chemical mutagenesis led to an overall improvement in L. rhamnosus growth and survival at a high concentration of aspirin (8mg/mL). The average beta-galactivity of unexposed cells was 153 +/- 2.5 Gal U. However, the mutant had a higher amount of beta-gal activity (169 +/- 1.5 Gal U). The mutant produced beta-gal when exposed to 4mg/ml of aspirin.The regular intake of medical drugs such as aspirin inhibits the Beta-Galactosidase activity and reduces protein expression a beneficial strain of bacteria exposed to aspirin, suggesting that this could occur in the gut microflora. The effect aspirin could have on gastrointestinal health should be taken into account when recommending it to users.
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773 0 $t Dissertation Abstracts International $g 78-12B(E).
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