Isolation and Identification of Waterborne Antibiotic-Resistant Bacteria and Molecular Characterization of their Antibiotic Resistance Genes

Devika Ghadigaonkar; Archana Rath

doi:10.3791/63934

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Isolation and Identification of Waterborne Antibiotic-Resistant Bacteria and Molecular Characterization of their Antibiotic Resistance Genes

水性抗生素耐药细菌的分离和鉴定及其抗生素耐药基因的分子表征

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08:58 min

March 3, 2023

DOI: 10.3791/63934-v

Devika Ghadigaonkar¹, Archana Rath¹

¹Antimicrobial Research (AMR) Lab, Department of Biotechnology,University of Mumbai

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Overview

This article presents a protocol for isolating and identifying antibiotic-resistant bacteria from water, along with the molecular characterization of their antibiotic resistance genes (ARGs). The combination of culture-based and non-culture-based techniques allows for comprehensive analysis of bacterial diversity and ARGs in freshwater samples from Mumbai, India.

Key Study Components

Area of Science

Microbiology
Environmental Science
Antibiotic Resistance

Background

Antibiotic-resistant bacteria pose a significant public health risk.
Water sources can harbor diverse bacterial populations, including resistant strains.
Understanding the prevalence and types of ARGs is crucial for managing antibiotic resistance.
Current methods may overlook non-culturable bacteria, which are significant in environmental samples.

Purpose of Study

To determine the overall load of antibiotic-resistant bacteria in water samples.
To identify the bacterial species present and their associated resistance traits.
To develop a replicable protocol applicable to various water sources.

Methods Used

Culture-based techniques for isolating bacteria.
Non-culture-based metagenomic analysis for comprehensive profiling.
Sample collection from diverse sources including sewage and clinical settings.
Characterization of antibiotic resistance genes through molecular techniques.

Main Results

The protocol successfully detects both culturable and non-culturable bacteria.
Identified a wide range of antibiotic resistance genes in the samples.
Demonstrated the effectiveness of combining different analytical techniques.
Provided insights into the diversity of antibiotic-resistant bacteria in freshwater.

Conclusions

The study highlights the importance of comprehensive methods in identifying antibiotic resistance.
Results can inform public health strategies for managing water quality.
The protocol can be adapted for various environmental and clinical applications.

Frequently Asked Questions

What types of samples can this protocol be applied to?

The protocol can be applied to samples from sewage, pharmaceutical effluents, hospital waste, and other clinical and nonclinical settings.

How does the method ensure no bacteria are left out?

By combining culture-based and non-culture-based techniques, the method can detect bacteria that are not easily grown in laboratory conditions.

What is the significance of identifying antibiotic resistance genes?

Identifying ARGs helps in understanding the mechanisms of resistance and can guide treatment and prevention strategies.

Who are the contributors to this study?

The study is conducted by researchers including Devika, Harshali Shinde, and Maitri Mishra from the laboratory.

What are the advantages of using metagenomic analysis?

Metagenomic analysis allows for the detection of a broader range of bacterial diversity and resistance genes, including those from non-culturable bacteria.

在这里，我们提出了一个详细的方案，用于从水中分离和鉴定抗生素耐药细菌及其抗生素抗性基因（ARG）的分子特征。使用基于培养物和非基于培养物（宏基因组分析）技术提供了有关印度孟买淡水中存在的细菌多样性和不同ARG的总库的完整信息。

我们的方法回答了关键问题，例如水样中抗生素耐药细菌的总体负荷是多少？这些细菌的身份是什么，它们携带的抗生素耐药性状和基因是什么？我们方案的优点是，它甚至可以检测出无法在实验室中生长的细菌，这些细菌占水样中细菌总数的很大一部分。

因此，从理论上讲，没有遗漏任何细菌或其耐药性状。这种涉及基于培养和独立于培养的技术的组合技术可以复制和定制从污水、制药或医院废水、临床和非临床环境等获得的任何样品。与Devika一起展示实验程序的是Harshali Shinde和Maitri Mishra，他们都是我实验室的高级研究员。

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