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埃塞俄比亚发酵饮料和食用鱼中分离出的乳酸菌对致病性和食源性致病微生物的体外拮抗作用

In Vitro Antagonistic Effect of Lactic Acid Bacteria Isolated from Fermented Beverage and Finfish on Pathogenic and Foodborne Pathogenic Microorganism in Ethiopia.

作者信息

Dejene Fitsum, Regasa Dadi Belayneh, Tadesse Dagimawie

机构信息

Department of Medical Microbiology, Arba Minch University, Arba Minch, Ethiopia.

出版信息

Int J Microbiol. 2021 Oct 15;2021:5370556. doi: 10.1155/2021/5370556. eCollection 2021.

Abstract

BACKGROUND

Lactic acid bacteria from fermented foods and fish can antagonistically inhibit the growth of foodborne pathogenic organism in fermented food and they stimulate the immune response to protect the fish from certain kinds of infections. The aim of this study was to evaluate the in vitro antagonistic activities of lactic acid bacteria isolated from fermented beverage (Borde) and finfish on foodborne pathogenic microorganisms.

METHODS

Laboratory-based experimental study was conducted from May 1 to Sep 1, 2020. Total sample numbers were 60 samples of fermented beverage (Borde) and 20 of finfish which were collected from different households and Chamo Lake (Arba Minch, Ethiopia). Each sample was firstly homogenized and serial dilution was prepared and spread on MRS agar plates in order to isolate pure culture. Different biochemical tests were performed to identify isolated bacteria. Then, cell-free supernatant (CFS) was prepared from MRS culture and used in an antimicrobial assay that was performed by agar diffusion method. The effects of pH, temperature, and enzymes on antimicrobial activity were evaluated in the same test. Simultaneously, the effects of lactic acid bacteria on aflatoxin production and on the permeability of the membrane were also evaluated. Data were analyzed using one-way ANOVA and Tukey post hoc analysis was performed by SPSS 25 statistical software.

RESULT

A total of 40 lactic acid bacteria were isolated; among them, 4 lactic acid bacteria, belonging to the genera , , and from fermented beverage and from fish, were screened for antimicrobial activity. The cell-free supernatant of those four isolates exhibited a significant ( < 0.05) antibacterial effect against tested pathogens and foodborne pathogenic bacteria. In addition, CFS showed antifungal and antiaflatoxigenic activities. The antimicrobial compounds synthesized by these isolates were sensitive to some proteolytic enzymes, and they were proved to be stable at high temperatures. It maintained/retained antimicrobial activity in a wide range of pH 2.0-10. Enterococcal CFS exhibited antibacterial activity against on membrane permeability, as confirmed by the increase in absorbance value between 0.075 and 0.24 at OD and between 0.68 and 1.2 at OD.

CONCLUSION

Cell-free supernatant produced by isolated lactic acid bacteria showed antimicrobial activity against a wide range of Gram-positive and Gram-negative foodborne bacteria, suggesting its potential application as a natural antimicrobial agent in tackling the rising drug resistance against foodborne pathogens.

摘要

背景

来自发酵食品和鱼类的乳酸菌可拮抗抑制发酵食品中食源性病原体的生长,并刺激免疫反应以保护鱼类免受某些感染。本研究的目的是评估从发酵饮料(博德)和有鳍鱼类中分离出的乳酸菌对食源性病原体微生物的体外拮抗活性。

方法

于2020年5月1日至9月1日进行了基于实验室的实验研究。总共采集了60份发酵饮料(博德)样本和20份有鳍鱼类样本,分别来自不同家庭和查莫湖(埃塞俄比亚阿尔巴明奇)。每个样本首先进行均质化处理,制备系列稀释液并涂布于MRS琼脂平板上以分离纯培养物。进行不同的生化试验以鉴定分离出的细菌。然后,从MRS培养物中制备无细胞上清液(CFS),并用于通过琼脂扩散法进行的抗菌测定。在同一试验中评估了pH、温度和酶对抗菌活性的影响。同时,还评估了乳酸菌对黄曲霉毒素产生和膜通透性的影响。数据使用单因素方差分析进行分析,并通过SPSS 25统计软件进行Tukey事后分析。

结果

共分离出40株乳酸菌;其中,对来自发酵饮料的4株乳酸菌(分别属于、和属)以及来自鱼类的1株乳酸菌进行了抗菌活性筛选。这四株分离株的无细胞上清液对受试病原体和食源性病原体细菌表现出显著(<0.05)的抗菌作用。此外,CFS显示出抗真菌和抗黄曲霉毒素活性。这些分离株合成的抗菌化合物对一些蛋白水解酶敏感,并且被证明在高温下稳定。它在pH 2.0 - 10的广泛范围内保持抗菌活性。肠球菌CFS对膜通透性表现出抗菌活性,通过OD处吸光度值在0.075至0.24之间以及OD处吸光度值在0.68至1.2之间的增加得到证实。

结论

分离出的乳酸菌产生的无细胞上清液对多种革兰氏阳性和革兰氏阴性食源细菌表现出抗菌活性,表明其作为天然抗菌剂在应对食源性病原体耐药性上升方面具有潜在应用价值。

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