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Tskhay, F.

Publications and source records attributed to Tskhay, F..

4 recordsLinked to original sources

Cycles of contamination and recovery: Combined sewer overflows drive acute but transient antimicrobial resistance exposure in an urban stream

Combined sewer overflows (CSOs) are a major pathway for untreated wastewater into urban streams, yet their role in shaping antimicrobial resistance (AMR) dynamics remains poorly understood. Here, we used high-frequency, time-resolved sampling during two storm-triggered CSO events across two monitoring locations and one stormwater-only control site in an urban stream to quantify how these disturbances affect microbial communities, antibiotic resistance genes (ARGs), and mobile genetic elements (MGEs) in an urban stream. CSO events caused rapid, up to two orders of magnitude, increases in bacterial, pathogen, and ARG abundance, with multiple transient peaks occurring within single overflow episodes. However, these increases were largely proportional to the total bacterial load, and most ARGs and MGEs did not change in relative abundance, indicating that CSOs primarily act as mass-transfer events rather than drivers of in situ selection. Downstream attenuation was governed by hydrological dilution despite additional CSO inputs: Both microbial and resistance signals largely returned to baseline within short time frames. This demonstrates that CSOs function as hydrologically driven pulse disturbances that generate acute but transient AMR exposure. Because CSO events lack the sustained pressure associated with continuous wastewater discharges, rapid washout prevents the long-term establishment of sewage-derived resistance. These findings highlight that AMR risk in CSO-impacted systems is driven primarily by short-term exposure rather than by persistent ecological transformation, with important implications for urban water management under increasingly extreme rainfall conditions.

microbiology↗

Plasmid-plasmid interactions reshape extracellular vesicle cargo and gene transfer potential

Plasmids are key drivers of horizontal gene transfer, yet their dissemination is not limited to conjugation. Extracellular vesicles (EVs) can transport plasmid DNA, but the factors governing plasmid incorporation into EVs remain poorly understood. Here, we tested whether principles of conjugative plasmid transfer, including plasmid mobility type and plasmid-plasmid interactions, extend to EV-mediated export. Using a conjugative plasmid (pKJK5) and a mobilizable plasmid (RSF1010) in two Gram-negative hosts, we quantified plasmid incorporation into EVs under single- and dual-plasmid conditions. When present individually, the conjugative plasmid was preferentially incorporated into EVs, exceeding RSF1010 by 10-23-fold despite its lower intracellular abundance. Under co-residence, this pattern reversed: RSF1010 became enriched by 13-39-fold, while pKJK5 was reduced by 2-7-fold. Consequently, EV-associated plasmid cargo shifted to RSF1010 dominance, deviating strongly from the expected 10-fold higher pKJK5 cargo if a stochastic model based on intracellular abundance and single-plasmid conditions were applicable. We propose that mobilizable plasmids under coexistence exploit conjugative plasmid transfer machinery to access membrane-associated sites, increasing their likelihood of incorporation into EVs. Our findings demonstrate that plasmid-plasmid interactions reshape EV cargo and identify a previously unrecognized mechanism that may influence extracellular gene transfer potential in microbial communities.

microbiology↗

Environmental reservoir of resistance genes for the last resort antibiotic Cefiderocol.

Antibiotic resistance poses a global public health threat. Cefiderocol, a recently introduced siderophore cephalosporin, employs a "Trojan Horse" mechanism by exploiting bacterial iron uptake systems for cell entry. Yet, resistant clinical isolates are already observed in clinics and resistance mechanisms are difficult to characterize. Here, we applied functional metagenomics to identify cefiderocol resistance genes. Functional metagenomic DNA libraries from diverse environmental samples collected across several countries were expressed in a cefiderocol-sensitive Escherichia coli host. This yielded four resistant clones with DNA originating from wastewater or freshwater DNA libraries. The identified antibiotic resistance genes (ARGs) causing an increase in cefiderocol minimum inhibitory concentrations encoded for beta-lactamases (VEB-3, OXA-372 homolog and YbxI homolog) and a partial penicillin binding protein homolog. Three of four shared closest homologs in pathogenic bacteria. One ARG was associated with a mobile genetic element and was broadly distributed across all wastewater samples from every country surveyed. This study underscores the critical importance of environmental surveillance for ARGs, particularly for novel agents like cefiderocol with limited understanding of resistance mechanisms.

microbiology↗

Fish are poor sentinels for surveillance of riverine AMR

Effective surveillance of antimicrobial resistance (AMR) in the environment is crucial for assessing the human and animal health risk of AMR pollution. Wastewater treatment plants (WWTPs) are one of the main sources of AMR pollutants discharged into water bodies. One important factor for assessing the risks associated with such pollution is the colonization potential of the resistant bacteria (ARB) and resistance genes (ARGs) from the environment into human or animal microbiomes upon exposure. This study explores whether fish can act as sentinels for surveillance of AMR pollution in general and specifically the human colonization potential of ARB in rivers impacted by WWTP effluents. Two riverine fish species, Brown trout, and European bullhead, were sampled up- and downstream a German WWTP. The two fish species were chosen due to their different lifestyles: Trout are mainly actively swimming in the water phase, while bullheads are sedentary and river sediment-associated. The bacterial microbiomes and resistomes of fish gills, skin, and feces were compared with those of the respective river water and sediment up- and downstream of the WWTP. Microbiomes of both fish mirrored the changes in river water and sediment downstream of the WWTP, with significant shifts in bacterial community composition, particularly an increase in Proteobacteria and Verrucomicrobia. However, increases in ARG abundances observed in water and sediment downstream of the WWTP were not reflected in any of the fish-associated resistomes. This indicates that while the fish microbiome is sensitive to environmental changes, resistomes of poikilothermic animals such as fish are less responsive to colonization by ARB originating from WWTPs and may not serve as effective sentinels for assessing AMR pollution and colonization risks in freshwater environments. This study highlights the complexity of using wildlife as indicators for environmental AMR pollution and suggests that other species are better suited for surveillance efforts. HighlightsO_LIWWTP effluent affected downstream river microbiome composition of water and sediment C_LIO_LIWWTP effluent significantly increased the level of six tested ARGs downstream C_LIO_LIChanges in environmental microbiome composition were mirrored in trout and bullhead C_LIO_LINo effect of the WWTP effluent on resistomes and individual ARG abundance in fishes C_LI

microbiology↗