Biology subjects

Ruhal, R.

Publications and source records attributed to Ruhal, R..

3 recordsLinked to original sources

bioRxiv2026

High-level ciprofloxacin resistance in Pseudomonas aeruginosa is associated with genomic adaptation, collateral antibiotic susceptibility, and physiological trade-offs

Antimicrobial resistance in Pseudomonas aeruginosa has emerged as a major clinical concern. Ciprofloxacin has potent intrinsic activity against P. aeruginosa, but resistance to this antibiotic is increasingly reported in clinical settings. In this study, P. aeruginosa ATCC 27853 strain was exposed to gradually increasing ciprofloxacin concentrations for 50 passages ([~]100 days). The final selected resistant strain (CIP-R) with 256-fold of minimum inhibitory concentration was studied further. Whole-genome sequencing revealed 12 genomic alterations, including known mutations in the quinolone resistance-determining genes gyrA (Thr83Ile), parC (Thr177Asn), parE (Glu459Lys). A duplication mutation in nfxB (Tyr153_Gly154dup) was also observed. In addition to these genes, we observed mutations in pilA, tadB, psdR, NP446_RS32055 (TRAP transporter permease), multiple dppA3 variants, and a prophage-associated hypothetical gene (NP446_RS24255), which have not been reported previously. The gyrA Thr83Ile substitution was conserved in 86.38% of ciprofloxacin-resistant clinical isolates retrieved from the NCBI database. Resistance acquisition was accompanied by slower growth, impaired swimming and swarming motility, diminished surface attachment, reduced biofilm formation. The resistant strain has enhanced {beta}-lactamase activity, and resistance to levofloxacin, cefepime, and meropenem together with sensitivity to piperacillin-tazobactam and aztreonam. This study highlights the gyrA Thr83Ile mutation as a key genomic marker for molecular screening of ciprofloxacin resistance and reveals secondary adaptive trade-offs that can be targeted for clinical diagnostic and therapeutic decision-making. In conclusion, achieving high-level ciprofloxacin resistance in P. aeruginosa involves non-target-site genomic adaptations and physiological trade-offs beyond classical target mutations.

microbiology↗

bioRxiv2025

Acinetobacter spp. with lower susceptibility to quaternary ammonium compounds enriched in microbial communities of frequently used sinks

Sanitary environments that undergo frequent cleaning and disinfection may harbor microbial communities with potential health risks. While biofilms in healthcare settings are well studied, comparatively less is known about sink microbiomes in public and educational buildings, where hundreds of people may interact with shared sink fixtures. This study characterized the spatial and temporal heterogeneity of sink drain biofilm microbiomes in academic buildings. We sampled 16 sinks from two buildings (four floors each, with sinks closest and furthest to the bathroom entrance), which are cleaned daily with quaternary ammonium compound (QAC) disinfectants, during periods of low and high student traffic (during and after academic breaks, respectively) across winter, spring, and summer. We observed significant spatial and temporal variations in microbial assemblages. Individual sinks accounted for 43% (PERMANOVA, p < 0.0001) of the variation in microbial communities. Microbiomes in each building were dominated by two genera, which together accounted for 30% of the community composition: Acinetobacter and Enhydrobacter (also classified as Moraxella) in the newer building, and Sphingomonas and Mycobacterium in the older building. Acinetobacter abundance varied seasonally and showed higher relative abundance during periods of high traffic. Metagenomic analysis of selected sinks revealed a high prevalence of qac genes and metagenome-assembled genomes (MAGs) harboring antimicrobial resistance genes (ARGs) including A. parvus. Notably, 34-53% of qac genes were co-localized on contigs associated with mobile genetic elements. These findings suggest that disinfected sink drains serve as persistent reservoirs of diverse microorganisms and potentially mobile resistance elements. IMPORTANCESink drains are recognized environmental reservoirs for multidrug-resistant bacteria and have been linked to healthcare-associated outbreaks. In public and educational buildings, these microbiomes are shaped by frequent human activity, making them potential sources of exposure and contributors to the environmental dissemination of antibiotic resistance genes. Quaternary ammonium compound (QAC) disinfectants are widely used on surfaces; however, they can select for resistant taxa and co-select for antibiotic resistance. In this study, despite routine cleaning with QACs, public restroom drains remain colonized by resilient biofilms, posing a potential risk to multiple users. Additionally, factors such as human traffic and seasonal variation may influence drain usage and microbial community composition. Elucidating how seasonal dynamics and human activity shape sink biofilms is essential for understanding their role in the environmental transmission of antimicrobial resistance and informing mitigation strategies in nonclinical settings.

microbiology↗

bioRxiv2019

Comparison of biofilm dispersal approaches in Pseudomonas aeruginosa and evaluation of dispersed cells in acute infection mouse model

Biofilm dispersal is biologically significant process to fully understand its consequences during biofilm based infections. The mechanism for biofilm dispersal may involve response to environmental cues and changes in intracellular secondary messengers. Considering range of cues for biofilm dispersal, it is significant to study if dispersed cells generated by different methods have similar phenotype. In the present study, we have compared four type of biofilm dispersal in P. aeruginosa based in response to environment cues (starvation and nutrient rich), c-di-GMP knockdown and signaling molecule NO. We have determined their dispersal efficiency, susceptibility of dispersed cells for antibiotics, their transcriptomic changes compared to planktonic and biofilm. We also determined if dispersed cells can cause acute infections in mouse models. In vitro experiments showed that NO and c-di-GMP based dispersal methods had high biofilm reduction efficiency of 50 and 75% respectively, however, the nutrient induced dispersion showed low efficiency ({approx}30%) and more tolerance to colistin. We also showed that in vitro dispersed cells induced >10-fold transcriptomic expression of genes (at significance level of p < 0.005) related to efflux pumps (mexCD-oprJ), antibiotic resistance (arnDET) in dispersed cells induced by NO and nutrient change, while denitrification pathway and virulence (T3SS, VreI, T2SS) genes in all dispersed cells compared to planktonic and biofilm state. When the ability of the dispersed cells was tested in mouse model of lung infection, c-di-GMP and NO based dispersed cells displayed enhanced infections and haematogenous spread to liver and spleen and higher mortality. Although degree of immune response (cytokines) does not differ based on phenotypes inoculated in our experimental conditions. Based on our data most efficient dispersal methods increase murine mortality. This indicates their capability of making biofilm associated infection more complicated. Our data encourage to be more careful for studies suggesting biofilm dispersal during treatment of biofilm infections.

microbiology↗