Uncovering antibiotic resistance: extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa from dipteran flies in residential dumping and livestock environments

IntroductionPseudomonas aeruginosa poses challenges in clinical and environmental contexts due to its capacity to colonize natural ecosystems and antibiotic resistance. This study characterized P. aeruginosa harboured by Diptera flies collected from illegal residential dumping sites and livestock (c...

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Main Authors: Lara de Wet, Itumeleng Matle, Oriel Thekisoe, Kgaugelo E. Lekota, Tsepo Ramatla
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-06-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1586811/full
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author Lara de Wet
Itumeleng Matle
Oriel Thekisoe
Kgaugelo E. Lekota
Tsepo Ramatla
author_facet Lara de Wet
Itumeleng Matle
Oriel Thekisoe
Kgaugelo E. Lekota
Tsepo Ramatla
author_sort Lara de Wet
collection DOAJ
description IntroductionPseudomonas aeruginosa poses challenges in clinical and environmental contexts due to its capacity to colonize natural ecosystems and antibiotic resistance. This study characterized P. aeruginosa harboured by Diptera flies collected from illegal residential dumping sites and livestock (cattle, sheep, and goats) kraals in Potchefstroom, South Africa.MethodsThe P. aeruginosa isolates were characterized using classical microbiological tests and species-specific gyrase B gene PCR assay. Antibiotic resistance (AR) was assessed on the isolates using disc diffusion assay (DDA). Additionally, PCR screened six virulence genes (exoS, plcN, plcH, toxA, lasB, and algD) among the isolates. Whole genome sequencing (WGS) was employed to confirm the identity and determine antibiotic resistance genes (ARGs) on selected isolates.ResultsCulture-based and molecular assays showed that P. aeruginosa is prevalent in Diptera flies (Hemipyrellia spp., Synthesiomya spp., Chrysomya spp., Sarchophagidae spp., and Tabanus spp.) from livestock kraals (75%; n = 36/48) and dumping sites (48%; n = 23/48). The most detected virulent gene among the isolates was exoS (96.6%), followed by plcN and algD genes (83.1%), lasB (81.4%), toxA (76.3%), and plcH (47.5%). All P. aeruginosa isolates were resistant to metronidazole, sulphamethoxazole, cefazolin and amoxicillin based on DDA. The sulfonamide resistance sulI gene (88.1%) was the most detected ARG from the P. aeruginosa isolates, followed by acc(3)-IV (80.6%) coding for aminoglycoside. WGS revealed that P. aeruginosa isolates belong to the sequence type (ST3808), which is multidrug-resistant and contains ARGs for fosfomycin (fosA), ampicillin (blaOXA-50), chloramphenicol (catB7), beta-lactamase (blaPAO), and aminoglycoside (aph(3’)-IIb).DiscussionThis study isolated ESBL-producing P. aeruginosa from various Diptera fly species collected from livestock kraals and residential dumping sites. This bacterium is important to “One Health” due to its multidrug resistance character and zoonotic nature. As a result, it requires consolidated control and management policies from the environmental, veterinary, and human health sectors.
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spelling doaj-art-81a363adbdd347dd89393546afbb9a672025-08-20T02:01:54ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-06-011610.3389/fmicb.2025.15868111586811Uncovering antibiotic resistance: extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa from dipteran flies in residential dumping and livestock environmentsLara de Wet0Itumeleng Matle1Oriel Thekisoe2Kgaugelo E. Lekota3Tsepo Ramatla4Unit for Environmental Sciences and Management, North-West University, Potchefstroom, South AfricaBacteriology Division, Onderstepoort Veterinary Research, Agricultural Research Council, Onderstepoort, South AfricaUnit for Environmental Sciences and Management, North-West University, Potchefstroom, South AfricaUnit for Environmental Sciences and Management, North-West University, Potchefstroom, South AfricaDepartment of Life Sciences, Centre for Applied Food Safety and Biotechnology, Central University of Technology, Bloemfontein, South AfricaIntroductionPseudomonas aeruginosa poses challenges in clinical and environmental contexts due to its capacity to colonize natural ecosystems and antibiotic resistance. This study characterized P. aeruginosa harboured by Diptera flies collected from illegal residential dumping sites and livestock (cattle, sheep, and goats) kraals in Potchefstroom, South Africa.MethodsThe P. aeruginosa isolates were characterized using classical microbiological tests and species-specific gyrase B gene PCR assay. Antibiotic resistance (AR) was assessed on the isolates using disc diffusion assay (DDA). Additionally, PCR screened six virulence genes (exoS, plcN, plcH, toxA, lasB, and algD) among the isolates. Whole genome sequencing (WGS) was employed to confirm the identity and determine antibiotic resistance genes (ARGs) on selected isolates.ResultsCulture-based and molecular assays showed that P. aeruginosa is prevalent in Diptera flies (Hemipyrellia spp., Synthesiomya spp., Chrysomya spp., Sarchophagidae spp., and Tabanus spp.) from livestock kraals (75%; n = 36/48) and dumping sites (48%; n = 23/48). The most detected virulent gene among the isolates was exoS (96.6%), followed by plcN and algD genes (83.1%), lasB (81.4%), toxA (76.3%), and plcH (47.5%). All P. aeruginosa isolates were resistant to metronidazole, sulphamethoxazole, cefazolin and amoxicillin based on DDA. The sulfonamide resistance sulI gene (88.1%) was the most detected ARG from the P. aeruginosa isolates, followed by acc(3)-IV (80.6%) coding for aminoglycoside. WGS revealed that P. aeruginosa isolates belong to the sequence type (ST3808), which is multidrug-resistant and contains ARGs for fosfomycin (fosA), ampicillin (blaOXA-50), chloramphenicol (catB7), beta-lactamase (blaPAO), and aminoglycoside (aph(3’)-IIb).DiscussionThis study isolated ESBL-producing P. aeruginosa from various Diptera fly species collected from livestock kraals and residential dumping sites. This bacterium is important to “One Health” due to its multidrug resistance character and zoonotic nature. As a result, it requires consolidated control and management policies from the environmental, veterinary, and human health sectors.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1586811/fullDiptera fliesPseudomonas aeruginosaantibiotic resistancevirulence genesWGS
spellingShingle Lara de Wet
Itumeleng Matle
Oriel Thekisoe
Kgaugelo E. Lekota
Tsepo Ramatla
Uncovering antibiotic resistance: extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa from dipteran flies in residential dumping and livestock environments
Frontiers in Microbiology
Diptera flies
Pseudomonas aeruginosa
antibiotic resistance
virulence genes
WGS
title Uncovering antibiotic resistance: extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa from dipteran flies in residential dumping and livestock environments
title_full Uncovering antibiotic resistance: extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa from dipteran flies in residential dumping and livestock environments
title_fullStr Uncovering antibiotic resistance: extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa from dipteran flies in residential dumping and livestock environments
title_full_unstemmed Uncovering antibiotic resistance: extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa from dipteran flies in residential dumping and livestock environments
title_short Uncovering antibiotic resistance: extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa from dipteran flies in residential dumping and livestock environments
title_sort uncovering antibiotic resistance extended spectrum beta lactamase producing pseudomonas aeruginosa from dipteran flies in residential dumping and livestock environments
topic Diptera flies
Pseudomonas aeruginosa
antibiotic resistance
virulence genes
WGS
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1586811/full
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