Adaptation and mutation ability of Bacillus subtilis and Pseudomonas fluorescens vegetative cells under the effect of a pulsed light stress

Nguyen Bao Loc * and Nicorescu Irina

* Corresponding author (nbloc@ctu.edu.vn)

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Received: 07 Apr 2015
Accepted: 21 Dec 2015
Published: 30 Dec 2015
Title: Adaptation and mutation ability of Bacillus subtilis and Pseudomonas fluorescens vegetative cells under the effect of a pulsed light stress
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How to Cite
Lộc, N. B., & Irina, N. . (2015). Adaptation and mutation ability of Bacillus subtilis and Pseudomonas fluorescens vegetative cells under the effect of a pulsed light stress. CTU Journal of Science, (41), 21-29. Retrieved from https://ctujsvn.ctu.edu.vn/index.php/ctujsvn/article/view/2183
Issue
No. 41 (2015)
Section
Food Technology

Abstract

The main objectives of this work were: (a) to compare the inactivation mechanisms and (b) to discuss similarities and differences in the adaptation and mutation behaviours of two bacterial model systems (Bacillus subtilis and Pseudomonas fluorescens) under a Pulsed Light (PL)-induced stress. When adaptation ability was investigated, firstly cells were exposed to sublethal PL doses (2×10-5 or 0.06 J.cm-2) and then submitted to a lethal PL treatment of 0.3 or 0.5 J.cm-2. Antibiotic resistance assays were carried out on bacterial suspensions in exponential growth phase in order to determine their mutagenic ability. For that, the effect of three PL lethal doses (0.2, 0.3 and 0.4 J.cm-2) was investigated. Experimental results showed that a low-energy PL dose (0.06 J.cm-2) was sufficient to produce B. subtilis adaptation, leading to an enhanced resistance to a subsequent lethal treatment. Conversely, P. fluorescens was not able to adapt to sublethal PL doses and more, 1 log additional microbial reduction was found when applying a lethal treatment. Applying a 0.2 J.cm-2 PL dose strongly increased the number of P. fluorescens resistant mutants compared to non-treated cells, while the mutation frequency was not modified for B. subtilis. As a conclusion, this study showed that these two highly adaptable Gram negative and positive bacteria have developed different behaviour as a response to low-energy PL.
Keywords: Pulsed light, inactivation, B. subtilis, P. fluorescens, adaptation, mutation

Tóm tắt

Mục đích của nghiên cứu này là (a) so sánh cơ chế tiêu diệt vi sinh vật và (b) khảo sát khả năng đáp ứng và đột biến của 2 giống vi sinh vật (Bacillus subtilis và Pseudomonas fluorescens) dưới tác dụng của xung ánh sáng. Để đánh giá khả năng đáp ứng, đầu tiên tế bào được xử lý bằng xung ánh sáng với liều năng lượng thích nghi (2x10-5 hoặc 0,06 J.cm-2), sau đó những tế bào này được xử lý tiếp với liều năng lượng tiêu diệt 0,3 hoặc 0,5 J.cm-2. Thí nghiệm thử khả năng kháng kháng sinh được thực hiện trên huyền phù vi sinh vật ở giai đoạn phát triển logarit để xác định khả năng đột biến của các vi sinh vật này. Thí nghiệm được thực hiện với 3 mức độ năng lượng của xung ánh sáng (0,2; 0,3 và 0,4 J.cm-2). Kết quả thí nghiệm cho thấy ở mức độ năng lượng xử lý thấp (0,06 J.cm-2) làm tăng khả năng đáp ứng của vi khuẩn B. subtilis, giúp cho vi khuẩn này chống chịu tốt hơn với liều năng lượng tiêu diệt xử lý tiếp theo. Ngược lại, P. fluorescens không đáp ứng với liều năng lượng thấp và hơn thế, việc tiền xử lý bằng liều năng lượng thấp còn làm tăng mức độ nhạy cảm của loài vi khuẩn này với liều năng lượng cao tiếp theo. Năng lượng xử lý 0,2 J.cm-2 làm tăng đáng kể số lượng vi khuẩn P. fluorescens đột biến so với mẫu không xử lý, trong khi đó kết quả này không được thể hiện ở vi khuẩn B. subtilis. Tóm lại, kết quả thí nghiệm cho thấy cả hai loại vi khuẩn Gram âm và Gram dương trong thí nghiệm này đều có khả năng đáp ứng với phương pháp xử lý xung ánh sáng, tuy nhiên cơ chế đáp ứng của mỗi loại không giống nhau.
Từ khóa: xung ánh sáng, ức chế, B. subtilis, P. fluorescens, đáp ứng, đột biến

Article Details

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