From wastewater to resistance: characterization of multidrug-resistant bacteria and assessment of natural antimicrobial compounds

The development and spread of antibiotic resistance in wastewater pose significant threats to both the environment and public health. Bacteria harboring multiple antibiotic resistance genes (ARGs), including those associated with horizontal gene transfer (HGT), can serve as persistent reservoirs and...

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Main Authors: Mingyue Li, Angela Zhan, Tahira Tasneem Rahman, Tao Jiang, Liyuan Hou
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1612534/full
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author Mingyue Li
Mingyue Li
Mingyue Li
Angela Zhan
Angela Zhan
Angela Zhan
Tahira Tasneem Rahman
Tahira Tasneem Rahman
Tao Jiang
Liyuan Hou
Liyuan Hou
author_facet Mingyue Li
Mingyue Li
Mingyue Li
Angela Zhan
Angela Zhan
Angela Zhan
Tahira Tasneem Rahman
Tahira Tasneem Rahman
Tao Jiang
Liyuan Hou
Liyuan Hou
author_sort Mingyue Li
collection DOAJ
description The development and spread of antibiotic resistance in wastewater pose significant threats to both the environment and public health. Bacteria harboring multiple antibiotic resistance genes (ARGs), including those associated with horizontal gene transfer (HGT), can serve as persistent reservoirs and vectors for antimicrobial resistance in natural ecosystems. In this study, nine antibiotic-resistant bacterial strains (U1–U9) were isolated from a wastewater treatment plant (WWTP) effluent. The isolates were identified using 16S rRNA gene sequencing and whole-genome sequencing (WGS), and their antibiotic susceptibility profiles were evaluated. All isolates exhibited resistance to multiple antibiotics, and WGS revealed that U1, U2, U4, and U7 harbored diverse ARGs, including β-lactamase genes, efflux pumps, and resistance determinants for sulfonamides, tetracyclines, and, quinolones, confirming the presence of multidrug-resistant bacteria in WWTP effluent. Phylogenetic analysis classified them into Microbacterium spp. (Actinobacteria), Chryseobacterium spp. (Bacteroidetes), Lactococcus lactis spp. (Firmicutes), and Psychrobacter spp. (Proteobacteria). To explore mitigation strategies, eleven natural compounds were screened for their effects on cell growth, biofilm formation, and motility in selected multi-drug-resistant bacteria. Among the tested compounds, curcumin and emodin showed the most consistent inhibitory activity, particularly against Microbacterium spp. strains U1 and U2, and Lactococcus lactis sp. U4. In contrast, Chryseobacterium sp. U7, a Gram-negative strain, exhibited strong resistance to all tested natural compounds, highlighting the challenge of controlling Gram-negative ARBs in wastewater settings. These findings underscore the environmental risks posed by multidrug-resistant and HGT-associated ARG-harboring bacteria in WWTP effluent. They also demonstrate the potential of natural products, such as curcumin and emodin, as alternative or complementary agents for mitigating antibiotic resistance in water systems.
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spelling doaj-art-bfa60a0cd146458e87e5d60698fae63f2025-08-20T02:36:20ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-07-011610.3389/fmicb.2025.16125341612534From wastewater to resistance: characterization of multidrug-resistant bacteria and assessment of natural antimicrobial compoundsMingyue Li0Mingyue Li1Mingyue Li2Angela Zhan3Angela Zhan4Angela Zhan5Tahira Tasneem Rahman6Tahira Tasneem Rahman7Tao Jiang8Liyuan Hou9Liyuan Hou10Utah Water Research Laboratory, Logan, UT, United StatesDepartment of Civil and Environmental Engineering, Utah State University, Logan, UT, United StatesSchool of Resources and Environmental Engineering, Shandong University of Technology, Zibo, ChinaUtah Water Research Laboratory, Logan, UT, United StatesDepartment of Civil and Environmental Engineering, Utah State University, Logan, UT, United StatesLogan High School, Logan, UT, United StatesUtah Water Research Laboratory, Logan, UT, United StatesDepartment of Civil and Environmental Engineering, Utah State University, Logan, UT, United StatesDepartment of Environmental and Sustainable Engineering, University at Albany, State University of New York, Albany, NY, United StatesUtah Water Research Laboratory, Logan, UT, United StatesDepartment of Civil and Environmental Engineering, Utah State University, Logan, UT, United StatesThe development and spread of antibiotic resistance in wastewater pose significant threats to both the environment and public health. Bacteria harboring multiple antibiotic resistance genes (ARGs), including those associated with horizontal gene transfer (HGT), can serve as persistent reservoirs and vectors for antimicrobial resistance in natural ecosystems. In this study, nine antibiotic-resistant bacterial strains (U1–U9) were isolated from a wastewater treatment plant (WWTP) effluent. The isolates were identified using 16S rRNA gene sequencing and whole-genome sequencing (WGS), and their antibiotic susceptibility profiles were evaluated. All isolates exhibited resistance to multiple antibiotics, and WGS revealed that U1, U2, U4, and U7 harbored diverse ARGs, including β-lactamase genes, efflux pumps, and resistance determinants for sulfonamides, tetracyclines, and, quinolones, confirming the presence of multidrug-resistant bacteria in WWTP effluent. Phylogenetic analysis classified them into Microbacterium spp. (Actinobacteria), Chryseobacterium spp. (Bacteroidetes), Lactococcus lactis spp. (Firmicutes), and Psychrobacter spp. (Proteobacteria). To explore mitigation strategies, eleven natural compounds were screened for their effects on cell growth, biofilm formation, and motility in selected multi-drug-resistant bacteria. Among the tested compounds, curcumin and emodin showed the most consistent inhibitory activity, particularly against Microbacterium spp. strains U1 and U2, and Lactococcus lactis sp. U4. In contrast, Chryseobacterium sp. U7, a Gram-negative strain, exhibited strong resistance to all tested natural compounds, highlighting the challenge of controlling Gram-negative ARBs in wastewater settings. These findings underscore the environmental risks posed by multidrug-resistant and HGT-associated ARG-harboring bacteria in WWTP effluent. They also demonstrate the potential of natural products, such as curcumin and emodin, as alternative or complementary agents for mitigating antibiotic resistance in water systems.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1612534/fullnatural compoundscurcuminemodinantibiotic-resistant bacteriawastewater treatment plantwhole-genome sequencing
spellingShingle Mingyue Li
Mingyue Li
Mingyue Li
Angela Zhan
Angela Zhan
Angela Zhan
Tahira Tasneem Rahman
Tahira Tasneem Rahman
Tao Jiang
Liyuan Hou
Liyuan Hou
From wastewater to resistance: characterization of multidrug-resistant bacteria and assessment of natural antimicrobial compounds
Frontiers in Microbiology
natural compounds
curcumin
emodin
antibiotic-resistant bacteria
wastewater treatment plant
whole-genome sequencing
title From wastewater to resistance: characterization of multidrug-resistant bacteria and assessment of natural antimicrobial compounds
title_full From wastewater to resistance: characterization of multidrug-resistant bacteria and assessment of natural antimicrobial compounds
title_fullStr From wastewater to resistance: characterization of multidrug-resistant bacteria and assessment of natural antimicrobial compounds
title_full_unstemmed From wastewater to resistance: characterization of multidrug-resistant bacteria and assessment of natural antimicrobial compounds
title_short From wastewater to resistance: characterization of multidrug-resistant bacteria and assessment of natural antimicrobial compounds
title_sort from wastewater to resistance characterization of multidrug resistant bacteria and assessment of natural antimicrobial compounds
topic natural compounds
curcumin
emodin
antibiotic-resistant bacteria
wastewater treatment plant
whole-genome sequencing
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1612534/full
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