Bacterivorous Ciliate <i>Tetrahymena pyriformis</i> Facilitates <i>vanA</i> Antibiotic Resistance Gene Transfer in <i>Enterococcus faecalis</i>

<b>Background:</b> Wastewater treatment plants (WWTPs) are hotspots for the emergence and spread of antibiotic resistance genes (ARGs). In activated sludge treatment systems, bacterivorous protozoa play a crucial role in biological processes, yet their impact on the horizontal gene trans...

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Main Authors: Temilola O. Olanrewaju, James S. G. Dooley, Heather M. Coleman, Chris McGonigle, Joerg Arnscheidt
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Antibiotics
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Online Access:https://www.mdpi.com/2079-6382/14/5/448
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author Temilola O. Olanrewaju
James S. G. Dooley
Heather M. Coleman
Chris McGonigle
Joerg Arnscheidt
author_facet Temilola O. Olanrewaju
James S. G. Dooley
Heather M. Coleman
Chris McGonigle
Joerg Arnscheidt
author_sort Temilola O. Olanrewaju
collection DOAJ
description <b>Background:</b> Wastewater treatment plants (WWTPs) are hotspots for the emergence and spread of antibiotic resistance genes (ARGs). In activated sludge treatment systems, bacterivorous protozoa play a crucial role in biological processes, yet their impact on the horizontal gene transfer in Gram-positive enteric bacteria remains largely unexplored. This study investigated whether the ciliate <i>Tetrahymena pyriformis</i> facilitates the transfer of antibiotic resistance genes between <i>Enterococcus faecalis</i> strains. <b>Methods</b>: Conjugation assays were conducted under laboratory conditions using a <i>vanA</i>-carrying donor and a rifampicin-resistant recipient at an initial bacterial concentration of 10<sup>9</sup> CFU/mL and ciliate density of 10<sup>5</sup> N/mL. <b>Results</b>: Transconjugant numbers peaked at 2 h when experiments started with recipient bacteria harvested in the exponential growth phase, and at 24 h when bacteria were in the stationary phase. In both cases, <i>vanA</i> gene transfer frequency was highest at 24 h (10<sup>−4</sup>–10<sup>−5</sup> CFU/mL), and the presence of energy sources increased gene transfer frequency by one order of magnitude. <b>Conclusions</b>: These findings suggest that ciliate grazing may contribute to <i>vanA</i> gene transfer in WWTP effluents, potentially facilitating its dissemination among permissive bacteria. Given the ecological and public health risks associated with <i>vanA</i> gene persistence in wastewater systems, understanding protozoan-mediated gene transfer is crucial for mitigating the spread of antibiotic resistance in aquatic environments.
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spelling doaj-art-706ce6d3cb374ffeba4f55d295ee91932025-08-20T03:14:32ZengMDPI AGAntibiotics2079-63822025-04-0114544810.3390/antibiotics14050448Bacterivorous Ciliate <i>Tetrahymena pyriformis</i> Facilitates <i>vanA</i> Antibiotic Resistance Gene Transfer in <i>Enterococcus faecalis</i>Temilola O. Olanrewaju0James S. G. Dooley1Heather M. Coleman2Chris McGonigle3Joerg Arnscheidt4School of Geography and Environmental Sciences, Ulster University, Cromore Road, Coleraine BT52 1SA, UKNutrition Innovation Centre for Food and Health (NICHE), School of Biomedical Sciences, Ulster University, Cromore Road, Coleraine BT52 1SA, UKSchool of Pharmacy and Pharmaceutical Sciences, Ulster University, Cromore Road, Coleraine BT52 1SA, UKSchool of Geography and Environmental Sciences, Ulster University, Cromore Road, Coleraine BT52 1SA, UKSchool of Geography and Environmental Sciences, Ulster University, Cromore Road, Coleraine BT52 1SA, UK<b>Background:</b> Wastewater treatment plants (WWTPs) are hotspots for the emergence and spread of antibiotic resistance genes (ARGs). In activated sludge treatment systems, bacterivorous protozoa play a crucial role in biological processes, yet their impact on the horizontal gene transfer in Gram-positive enteric bacteria remains largely unexplored. This study investigated whether the ciliate <i>Tetrahymena pyriformis</i> facilitates the transfer of antibiotic resistance genes between <i>Enterococcus faecalis</i> strains. <b>Methods</b>: Conjugation assays were conducted under laboratory conditions using a <i>vanA</i>-carrying donor and a rifampicin-resistant recipient at an initial bacterial concentration of 10<sup>9</sup> CFU/mL and ciliate density of 10<sup>5</sup> N/mL. <b>Results</b>: Transconjugant numbers peaked at 2 h when experiments started with recipient bacteria harvested in the exponential growth phase, and at 24 h when bacteria were in the stationary phase. In both cases, <i>vanA</i> gene transfer frequency was highest at 24 h (10<sup>−4</sup>–10<sup>−5</sup> CFU/mL), and the presence of energy sources increased gene transfer frequency by one order of magnitude. <b>Conclusions</b>: These findings suggest that ciliate grazing may contribute to <i>vanA</i> gene transfer in WWTP effluents, potentially facilitating its dissemination among permissive bacteria. Given the ecological and public health risks associated with <i>vanA</i> gene persistence in wastewater systems, understanding protozoan-mediated gene transfer is crucial for mitigating the spread of antibiotic resistance in aquatic environments.https://www.mdpi.com/2079-6382/14/5/448antimicrobial resistanceantibiotic resistant bacteriahorizontal gene transferconjugationprotistsprotozoa
spellingShingle Temilola O. Olanrewaju
James S. G. Dooley
Heather M. Coleman
Chris McGonigle
Joerg Arnscheidt
Bacterivorous Ciliate <i>Tetrahymena pyriformis</i> Facilitates <i>vanA</i> Antibiotic Resistance Gene Transfer in <i>Enterococcus faecalis</i>
Antibiotics
antimicrobial resistance
antibiotic resistant bacteria
horizontal gene transfer
conjugation
protists
protozoa
title Bacterivorous Ciliate <i>Tetrahymena pyriformis</i> Facilitates <i>vanA</i> Antibiotic Resistance Gene Transfer in <i>Enterococcus faecalis</i>
title_full Bacterivorous Ciliate <i>Tetrahymena pyriformis</i> Facilitates <i>vanA</i> Antibiotic Resistance Gene Transfer in <i>Enterococcus faecalis</i>
title_fullStr Bacterivorous Ciliate <i>Tetrahymena pyriformis</i> Facilitates <i>vanA</i> Antibiotic Resistance Gene Transfer in <i>Enterococcus faecalis</i>
title_full_unstemmed Bacterivorous Ciliate <i>Tetrahymena pyriformis</i> Facilitates <i>vanA</i> Antibiotic Resistance Gene Transfer in <i>Enterococcus faecalis</i>
title_short Bacterivorous Ciliate <i>Tetrahymena pyriformis</i> Facilitates <i>vanA</i> Antibiotic Resistance Gene Transfer in <i>Enterococcus faecalis</i>
title_sort bacterivorous ciliate i tetrahymena pyriformis i facilitates i vana i antibiotic resistance gene transfer in i enterococcus faecalis i
topic antimicrobial resistance
antibiotic resistant bacteria
horizontal gene transfer
conjugation
protists
protozoa
url https://www.mdpi.com/2079-6382/14/5/448
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AT joergarnscheidt bacterivorousciliateitetrahymenapyriformisifacilitatesivanaiantibioticresistancegenetransferinienterococcusfaecalisi