Development of an in vitro biofilm model for the study of the impact of fluoroquinolones on sewer biofilm microbiota
Sewer biofilms are likely to constitute hotspots for selecting and accumulating antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs). This study aimed to optimize culture conditions to obtain in vitro biofilms, mimicking the biofilm collected in sewers, to study the impact of f...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2024-03-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1377047/full |
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| author | Sarah A. Naudin Aude A. Ferran Pedro Henrique Imazaki Nathalie Arpaillange Camille Marcuzzo Maïna Vienne Maïna Vienne Sofia Demmou Alain Bousquet-Mélou Felipe Ramon-Portugal Marlene Z. Lacroix Claire Hoede Claire Hoede Maialen Barret Véronique Dupouy Delphine Bibbal |
| author_facet | Sarah A. Naudin Aude A. Ferran Pedro Henrique Imazaki Nathalie Arpaillange Camille Marcuzzo Maïna Vienne Maïna Vienne Sofia Demmou Alain Bousquet-Mélou Felipe Ramon-Portugal Marlene Z. Lacroix Claire Hoede Claire Hoede Maialen Barret Véronique Dupouy Delphine Bibbal |
| author_sort | Sarah A. Naudin |
| collection | DOAJ |
| description | Sewer biofilms are likely to constitute hotspots for selecting and accumulating antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs). This study aimed to optimize culture conditions to obtain in vitro biofilms, mimicking the biofilm collected in sewers, to study the impact of fluoroquinolones (FQs) on sewer biofilm microbiota. Biofilms were grown on coupons in CDC Biofilm Reactors®, continuously fed with nutrients and inoculum (1/100 diluted wastewater). Different culture conditions were tested: (i) initial inoculum: diluted wastewater with or without sewer biofilm, (ii) coupon material: concrete vs. polycarbonate, and (iii) time of culture: 7 versus 14 days. This study found that the biomass was highest when in vitro biofilms were formed on concrete coupons. The biofilm taxonomic diversity was not affected by adding sewer biofilm to the initial inoculum nor by the coupon material. Pseudomonadales, Burkholderiales and Enterobacterales dominated in the sewer biofilm composition, whereas in vitro biofilms were mainly composed of Enterobacterales. The relative abundance of qnrA, B, D and S genes was higher in in vitro biofilms than sewer biofilm. The resistome of sewer biofilm showed the highest Shannon diversity index compared to wastewater and in vitro biofilms. A PCoA analysis showed differentiation of samples according to the nature of the sample, and a Procrustes analysis showed that the ARG changes observed were linked to changes in the microbial community. The following growing conditions were selected for in vitro biofilms: concrete coupons, initial inoculation with sewer biofilm, and a culture duration of 14 days. Then, biofilms were established under high and low concentrations of FQs to validate our in vitro biofilm model. Fluoroquinolone exposure had no significant impact on the abundance of qnr genes, but high concentration exposure increased the proportion of mutations in gyrA (codons S83L and D87N) and parC (codon S80I). In conclusion, this study allowed the determination of the culture conditions to develop an in vitro model of sewer biofilm; and was successfully used to investigate the impact of FQs on sewer microbiota. In the future, this setup could be used to clarify the role of sewer biofilms in disseminating resistance to FQs in the environment. |
| first_indexed | 2024-04-24T18:46:29Z |
| format | Article |
| id | doaj.art-99049cf001c643c98bbc3d34df739db3 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-24T18:46:29Z |
| publishDate | 2024-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-99049cf001c643c98bbc3d34df739db32024-03-27T05:11:45ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2024-03-011510.3389/fmicb.2024.13770471377047Development of an in vitro biofilm model for the study of the impact of fluoroquinolones on sewer biofilm microbiotaSarah A. Naudin0Aude A. Ferran1Pedro Henrique Imazaki2Nathalie Arpaillange3Camille Marcuzzo4Maïna Vienne5Maïna Vienne6Sofia Demmou7Alain Bousquet-Mélou8Felipe Ramon-Portugal9Marlene Z. Lacroix10Claire Hoede11Claire Hoede12Maialen Barret13Véronique Dupouy14Delphine Bibbal15INTHERES, Université de Toulouse, INRAE, ENVT, Toulouse, FranceINTHERES, Université de Toulouse, INRAE, ENVT, Toulouse, FranceINTHERES, Université de Toulouse, INRAE, ENVT, Toulouse, FranceINTHERES, Université de Toulouse, INRAE, ENVT, Toulouse, FranceINTHERES, Université de Toulouse, INRAE, ENVT, Toulouse, FranceUniversité de Toulouse, INRAE, BioinfOmics, GenoToul Bioinformatics Facility, Castanet-Tolosan, FranceUniversité de Toulouse, INRAE, UR 875 MIAT, Castanet-Tolosan, FranceCentre de Recherche sur la Biodiversité et l’Environnement, Université de Toulouse, CNRS, IRD, Toulouse INP, Université de Toulouse, Toulouse, FranceINTHERES, Université de Toulouse, INRAE, ENVT, Toulouse, FranceINTHERES, Université de Toulouse, INRAE, ENVT, Toulouse, FranceINTHERES, Université de Toulouse, INRAE, ENVT, Toulouse, FranceUniversité de Toulouse, INRAE, BioinfOmics, GenoToul Bioinformatics Facility, Castanet-Tolosan, FranceUniversité de Toulouse, INRAE, UR 875 MIAT, Castanet-Tolosan, FranceCentre de Recherche sur la Biodiversité et l’Environnement, Université de Toulouse, CNRS, IRD, Toulouse INP, Université de Toulouse, Toulouse, FranceINTHERES, Université de Toulouse, INRAE, ENVT, Toulouse, FranceINTHERES, Université de Toulouse, INRAE, ENVT, Toulouse, FranceSewer biofilms are likely to constitute hotspots for selecting and accumulating antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs). This study aimed to optimize culture conditions to obtain in vitro biofilms, mimicking the biofilm collected in sewers, to study the impact of fluoroquinolones (FQs) on sewer biofilm microbiota. Biofilms were grown on coupons in CDC Biofilm Reactors®, continuously fed with nutrients and inoculum (1/100 diluted wastewater). Different culture conditions were tested: (i) initial inoculum: diluted wastewater with or without sewer biofilm, (ii) coupon material: concrete vs. polycarbonate, and (iii) time of culture: 7 versus 14 days. This study found that the biomass was highest when in vitro biofilms were formed on concrete coupons. The biofilm taxonomic diversity was not affected by adding sewer biofilm to the initial inoculum nor by the coupon material. Pseudomonadales, Burkholderiales and Enterobacterales dominated in the sewer biofilm composition, whereas in vitro biofilms were mainly composed of Enterobacterales. The relative abundance of qnrA, B, D and S genes was higher in in vitro biofilms than sewer biofilm. The resistome of sewer biofilm showed the highest Shannon diversity index compared to wastewater and in vitro biofilms. A PCoA analysis showed differentiation of samples according to the nature of the sample, and a Procrustes analysis showed that the ARG changes observed were linked to changes in the microbial community. The following growing conditions were selected for in vitro biofilms: concrete coupons, initial inoculation with sewer biofilm, and a culture duration of 14 days. Then, biofilms were established under high and low concentrations of FQs to validate our in vitro biofilm model. Fluoroquinolone exposure had no significant impact on the abundance of qnr genes, but high concentration exposure increased the proportion of mutations in gyrA (codons S83L and D87N) and parC (codon S80I). In conclusion, this study allowed the determination of the culture conditions to develop an in vitro model of sewer biofilm; and was successfully used to investigate the impact of FQs on sewer microbiota. In the future, this setup could be used to clarify the role of sewer biofilms in disseminating resistance to FQs in the environment.https://www.frontiersin.org/articles/10.3389/fmicb.2024.1377047/fullwastewatersewerbiofilmfluoroquinolonebioreactorantibiotic resistance |
| spellingShingle | Sarah A. Naudin Aude A. Ferran Pedro Henrique Imazaki Nathalie Arpaillange Camille Marcuzzo Maïna Vienne Maïna Vienne Sofia Demmou Alain Bousquet-Mélou Felipe Ramon-Portugal Marlene Z. Lacroix Claire Hoede Claire Hoede Maialen Barret Véronique Dupouy Delphine Bibbal Development of an in vitro biofilm model for the study of the impact of fluoroquinolones on sewer biofilm microbiota Frontiers in Microbiology wastewater sewer biofilm fluoroquinolone bioreactor antibiotic resistance |
| title | Development of an in vitro biofilm model for the study of the impact of fluoroquinolones on sewer biofilm microbiota |
| title_full | Development of an in vitro biofilm model for the study of the impact of fluoroquinolones on sewer biofilm microbiota |
| title_fullStr | Development of an in vitro biofilm model for the study of the impact of fluoroquinolones on sewer biofilm microbiota |
| title_full_unstemmed | Development of an in vitro biofilm model for the study of the impact of fluoroquinolones on sewer biofilm microbiota |
| title_short | Development of an in vitro biofilm model for the study of the impact of fluoroquinolones on sewer biofilm microbiota |
| title_sort | development of an in vitro biofilm model for the study of the impact of fluoroquinolones on sewer biofilm microbiota |
| topic | wastewater sewer biofilm fluoroquinolone bioreactor antibiotic resistance |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1377047/full |
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