Ciprofloxacin Exposure Research Articles

Consumption of a Western-Style Diet Modulates the Response of the Murine Gut Microbiome to Ciprofloxacin.

Dietary composition and antibiotic use have major impacts on the structure and function of the gut microbiome, often resulting in dysbiosis. Despite this, little research has been done to explore the role of host diet as a determinant of antibiotic-induced microbiome disruption. Here, we utilize a multi-omic approach to characterize the impact of Western-style diet consumption on ciprofloxacin-induced changes to gut microbiome structure and transcriptional activity. We found that Western diet consumption dramatically increased Bacteroides abundances and shifted the community toward the metabolism of simple sugars and mucus glycoproteins. Mice consuming a Western-style diet experienced a greater expansion of Firmicutes following ciprofloxacin treatment than those eating a control diet. Transcriptionally, we found that ciprofloxacin reduced the abundance of tricarboxylic acid (TCA) cycle transcripts on both diets, suggesting that carbon metabolism plays a key role in the response of the gut microbiome to this antibiotic. Despite this, we observed extensive diet-dependent differences in the impact of ciprofloxacin on microbiota function. In particular, at the whole-community level we detected an increase in starch degradation, glycolysis, and pyruvate fermentation following antibiotic treatment in mice on the Western diet, which we did not observe in mice on the control diet. Similarly, we observed diet-specific changes in the transcriptional activity of two important commensal bacteria, Akkermansia muciniphila and Bacteroides thetaiotaomicron, involving diverse cellular processes such as nutrient acquisition, stress responses, and capsular polysaccharide (CPS) biosynthesis. These findings demonstrate that host diet plays a role in determining the impacts of ciprofloxacin on microbiome composition and microbiome function.IMPORTANCE Due to the growing incidence of disorders related to antibiotic-induced dysbiosis, it is essential to determine how our "Western"-style diet impacts the response of the microbiome to antibiotics. While diet and antibiotics have profound impacts on gut microbiome composition, little work has been done to examine their combined effects. Previous work has shown that nutrient availability, influenced by diet, plays an important role in determining the extent of antibiotic-induced disruption to the gut microbiome. Thus, we hypothesize that the Western diet will shift microbiota metabolism toward simple sugar and mucus degradation and away from polysaccharide utilization. Because of bacterial metabolism's critical role in antibiotic susceptibility, this change in baseline metabolism will impact how the structure and function of the microbiome are impacted by ciprofloxacin exposure. Understanding how diet modulates antibiotic-induced microbiome disruption will allow for the development of dietary interventions that can alleviate many of the microbiome-dependent complications of antibiotic treatment.

Ciprofloxacin-induced persister-cells in Campylobacter jejuni.

Campylobacter jejuni is a major bacterial foodborne-pathogen. Ciprofloxacin is an important antibiotic for the treatment of C. jejuni, albeit high rates of fluoroquinolone resistance have limited its usefulness. Persister-cells are transiently antibiotic-tolerant fractions of bacterial populations and their occurrence has been associated with recalcitrant and persistent bacterial infections. Here, time-kill assays with ciprofloxacin (200×MIC, 25 µg ml-1) were performed in C. jejuni strains 81-176 and RM1221 and persister-cells were found. The frequency of survivors after 8 h of ciprofloxacin exposure was approx. 10-3 for both strains, while after 22 h the frequency was between 10-5-10-7, depending on the strain and growth-phase. Interestingly, the stationary-phase cultures did not display more persister-cells compared to exponential-phase cultures, in contrast to what has been observed in other bacterial species. Persister-cells after ampicillin exposure (100×MIC, 200 µg ml-1) were not detected, implying that persister-cell formation in C. jejuni is antibiotic-specific. In attempts to identify the mechanism of ciprofloxacin persister-cell formation, stringent or SOS responses were not found to play major roles. Overall, this study reports ciprofloxacin persister-cells in C. jejuni and challenges the notion of persister-cells as plainly dormant non-growing cells.

Oxytetracycline and Ciprofloxacin Exposure Altered the Composition of Protistan Consumers in an Agricultural Soil.

Protists, an integral component of soil microbiome, are one of the main predators of bacteria. Bacteria can produce toxic secondary metabolites, e.g., antibiotics to fight stress under the predation pressure of protists; however, impacts of antibiotics on the profile of protists in soils remain unclear. Here, we constructed a microcosm incubation to investigate the effects of two common antibiotics, oxytetracycline and ciprofloxacin, on the protistan and bacterial communities in an arable soil. Rhizaria were the most abundant protist supergroup, followed by Amoebozoa, Stramenopiles, and Aveolata. Among trophic functional groups, consumers were predominant within the protistan community. The protistan alpha-diversity was not significantly changed, while the bacterial alpha-diversity was decreased under the pressure of antibiotics. Nevertheless, the antibiotic exposure considerably reduced the relative abundance of protistan lineages in Rhizaria and Amoebozoa, which were the dominant supergroups of protistan consumers, while increased the relative abundance of other consumer and phototrophic protists. Altogether, we provide novel experimental evidence that the bacterivorous consumers, an important functional group of protists, were more sensitive to antibiotics than other functional groups. Our findings have potential implications for the induced alterations of protistan community and their ecological functions under the scenarios of projected increasing global antibiotic usage.

Overexpression of RamA, Which Regulates Production of the Multidrug Resistance Efflux Pump AcrAB-TolC, Increases Mutation Rate and Influences Drug Resistance Phenotype.

In Enterobacteriales, the AcrAB-TolC efflux pump exports substrates, including antimicrobials, from the cell. Overexpression of AcrAB-TolC can occur after exposure to fluoroquinolones, leading to multidrug resistance. The expression of AcrAB-TolC in Salmonella is primarily regulated by the transcriptional activator RamA. However, other transcriptional activators, such as MarA, SoxRS, and Rob, can influence AcrAB-TolC expression. This study determined whether the overproduction or absence of RamA influences the mutation rate or the phenotype of mutants selected in Salmonella enterica serovar Typhimurium SL1344 after ciprofloxacin exposure. The absence of RamA (SL1344 ramA::aph) resulted in mutation frequencies/rates similar to those of wild-type Salmonella Typhimurium SL1344. However, the overproduction of RamA (SL1344 ramR::aph) and, consequently, AcrB resulted in a significantly higher mutation frequency and rate than for wild-type Salmonella Typhimurium SL1344. Whole-genome sequencing revealed that in addition to selecting gyrA mutants resistant to quinolones, SL1344 and SL1344 ramA::aph also produced multidrug-resistant (MDR) mutants, associated with mutations in soxR Conversely, mutations in SL1344 ramR::aph occurred in gyrA only. Although transcriptional regulators such as SoxRS are believed to play a minor role in AcrAB-TolC regulation under antibiotic selective pressure, we show that soxR mutants can be selected after exposure to ciprofloxacin, including when RamA is absent. This demonstrates that under selective pressure, Salmonella can respond to increased efflux pump expression by mutating other AcrAB-TolC regulatory genes, allowing for the evolution of MDR. Understanding how Salmonella responds to antibiotic pressure in the absence/overproduction of RamA is important if targeting transcriptional regulators to alter efflux is to be considered an avenue for future drug discovery.

Effects of ciprofloxacin exposure on the earthworm Eisenia fetida

The widespread use of the antibiotic ciprofloxacin (CIP) poses a serious risk to soil organisms. Here, earthworms (Eisenia fetida) were used to explore the effect of CIP exposure on growth, reproduction, mortality, antioxidant enzyme activity, DNA damage, and mRNA levels. The results showed that mortality, weight, and reproduction did not change in response to CIP exposure. The antioxidant enzymes superoxide dismutase (SOD), peroxidase (POD), and glutathione (GSH) were inhibited after 10 mg/kg CIP exposure on day 21, and catalase (CAT) activity increased significantly on day 14. High concentrations (1–2 g/kg) of CIP pollution caused DNA damage in E. fetida on days 7 and 14. At a dose of 10 mg/kg, CIP altered antioxidant enzymes and gene expression, but was not harmful to the growth and reproduction of E. fetida. Moreover, mRNA expression of annetocin, metallothionein, heat shock protein 70, and translationally controlled tumor protein were significantly upregulated on day 28. These findings indicate that antioxidant enzymes, DNA damage, and mRNA levels of biomarkers are more sensitive than mortality, growth, and reproduction for detecting CIP pollution in the soil environment.

The Effect of Ciprofloxacin Exposure on Ctx-M Gene Pattern of Escherichia Coli

Background: An exposure of ciprofloxacin on Escherichia coli (E.coli) may cause cross-resistance to beta-lactam groups. The researchers obtained that ciprofloxacin, aminoglycoside, cotrimoxazole and second generation of cephalosporins were the risk factors for infection caused by Extended-Spectrum ?-Lactamases (ESBL)-producing bacteria. Exposure of ciprofloxacin to E.coli causes resistance of cefotaxime and produces ESBL which is genotypically evidenced by the presence of CTX-M.Objective: To know the effect of ciprofloxacin exposure on CTX-M gene pattern of E. coli.Method: A total of 30 E.coli isolates were exposed to ciprofloxacin for 14 days with the Kirby-Bauer antibiotic disc diffusion method. ESBL confirmatory test and Modified Double Disk Susceptibility Test (MDDST) method was used the antibiotic disc and continued electrophoresis using Polymerase Chain Reaction (PCR). The data analysis used the chi-square statistic test with ? = 0.05.Results: In the PCR examination, 10.00% of ESBL isolates were obtained to have CTX-M-15 gene, and 33.00% of non-ESBL E.coli isolates having CTX-M-15 gene. While 10.00% of E.coli ESBL isolates and 53.30% non-ESBL did not have CTX-M-15 genes. There was no significant difference between ciprofloxacin exposure and the presence of CTX-M-15 gene (p = 0.290).Conclusion: Most of the E.coli isolates had CTX-M-15 gene after the exposure. There was no difference in ciprofloxacin exposure to the presence of CTX-M-15 genes of E. coli.

Chronic ecotoxicity of ciprofloxacin exposure on taxonomic diversity of a meiobenthic nematode community in microcosm experiments

A laboratory bioassay was performed to assess the ecotoxicity of ciprofloxacin on a meiobenthic nematodes community from Bizerte lagoon (NE Tunisia). Four ciprofloxacin doses [D1 (50 µg/g), D2 (100 µg/g), D3 (200 µg/g), and D4 (500 µg/g)] were applied to the substrate, and responses were studied after one month. Discernible differences were observed between control assemblages and those populating ciprofloxacin treated substrates. All univariate indices were affected significantly compared to those in the control microcosm with increasing antibiotic concentration. The non-parametric Multi-Dimensional Scaling based on species abundances (MDS) showed significant separation of the control microcosm from the antibiotic-treated populations. The nematode species responses to the fluoroquinolone treatments varied: Odontophora villoti was reduced at all concentrations of ciprofloxacin and was considered “sensitive,” whereas Metoncholaimus pristiurus was affected by moderate concentrations; its abundance increased with the highest dose D4 and was described as “opportunistic.” Paramonohystera pilosa, whose abundance increased with antibiotic doses appeared “resistant.”

Migration of antibiotic ciprofloxacin during phytoremediation of contaminated water and identification of transformation products

Phytoremediation is an effective and environmentally friendly approach to treat antibiotic contaminated water, however, the mechanisms of migration and transformation of antibiotics in plant tissues are still far from clear. In this study, the floating macrophyte Eichhornia crassipes was exposed to a series of antibiotic ciprofloxacin (CIP) concentrations. The results showed that the CIP was taken up and accumulated in the roots, which were the major accumulative tissue. CIP content increased with lipid content. During cultivation, the root bioconcentration factor (RCF) gradually increased. The average CIP content detected in aerial parts was 12.80 μg g−1, an order of magnitude lower than in the roots. At low CIP concentrations, the highest leaf bioconcentration factor (LCF) and transfer factor (TF) indicated highly efficient translocation from roots to aerial parts. The soluble protein growth rate of leaves, which is associated with metabolic activity, increased following CIP exposure. Overall, eight major transformation products in E. crassipes tissues were identified, and three possible transformation pathways were proposed involving the processes of desethylation, dehydroxylation, oxidation, hydroxylation and cleavage of the piperazine and quinoline rings. These findings could prove beneficial for improving the management or amelioration methods used for treating water contaminated with antibiotics.

Population Pharmacokinetics and Pharmacodynamics of Ciprofloxacin Prophylaxis in Pediatric Acute Lymphoblastic Leukemia Patients.

Ciprofloxacin is used as antimicrobial prophylaxis in pediatric acute lymphoblastic leukemia (ALL) to decrease infections with gram-negative bacteria. However, there are no clear guidelines concerning prophylactic dose. To determine the pharmacokinetics and pharmacodynamics (PKPD) of ciprofloxacin prophylaxis in a pediatric ALL population. The effect of patient characteristics and antileukemic treatment on ciprofloxacin exposure, the area under the concentration time curve over minimal inhibitory concentration (AUC24/MIC) ratios, and emergence of resistance were studied. A total of 615 samples from 129 children (0-18 years) with ALL were collected in a multicenter prospective study. A population pharmacokinetic model was developed. Microbiological cultures were collected prior to and during prophylaxis. An AUC24/MIC of ≥125 was defined as target ratio. A 1-compartment model with zero-order absorption and allometric scaling best described the data. No significant (P < .01) covariates remained after backward elimination and no effect of asparaginase or azoles were found. Ciprofloxacin AUC24 was 16.9 mg*h/L in the prednisone prophase versus 29.3 mg*h/L with concomitant chemotherapy. Overall, 100%, 81%, and 18% of patients at, respectively, MIC of 0.063, 0.125, and 0.25 mg/L achieved AUC24/MIC ≥ 125. In 13% of the patients, resistant bacteria were found during prophylactic treatment. Ciprofloxacin exposure shows an almost 2-fold change throughout the treatment of pediatric ALL. Depending on the appropriateness of 125 as target ratio, therapeutic drug monitoring or dose adjustments might be indicated for less susceptible bacteria starting from ≥ 0.125 mg/L to prevent the emergence of resistance and reach required targets for efficacy.

Effects of ciprofloxacin on metabolic activity and algal biomass of urban stream biofilms

Pharmaceuticals and personal care products (PPCPs), such as the commonly prescribed antibiotic ciprofloxacin, are present and persistent in freshwaters, yet their effects on aquatic ecosystem functions at environmentally-relevant concentrations are rarely explored. Stream biofilms provide multiple functions in stream ecosystems, but their functional response to PPCP contaminants such as ciprofloxacin is unclear. To establish the effect of ciprofloxacin on aquatic biofilms, we colonized biofilms in situ on tiles (n = 80) at four sites along an urban stream in Gainesville, Florida, including two sites above and two sites below a wastewater treatment plant (WWTP). We then incubated the tiles and associated biofilms in the laboratory for 6 d exposing biofilms to either 0, 0.01, 0.1, or 1.0 μg/L (target concentrations) of ciprofloxacin. At the end of the 6 d laboratory exposure, we quantified gross primary production (GPP), respiration (R), and biomass (as chlorophyll a) of biofilms, and calculated response ratios for each response. All response metrics were significantly differed across sites (p < 0.01). Ciprofloxacin significantly decreased GPP (p < 0.05) regardless of treatment concentration, most notably at the site immediately below the WWTP, where there was no measurable GPP on any ciprofloxacin-treated biofilms. In contrast, respiration (R) was not significantly affected by ciprofloxacin, despite an apparent increase in R at the WWTP site. However, the WWTP site R was significantly different from the most upstream and downstream sites (p < 0.001) but was not significantly different from a nearby site upstream of the WWTP (p > 0.05). These results indicate that chronic exposure to ciprofloxacin through WWTP effluent can alter ecosystem functions performed by biofilms, which can have consequences for higher trophic levels and stream processes. By quantifying biofilm metabolic responses to ciprofloxacin exposure, this study supports the concept that pharmaceuticals and personal care products can induce sub-lethal effects on ecological processes at environmentally-relevant concentrations.

Understanding Ciprofloxacin Failure in Pseudomonas aeruginosa Biofilm: Persister Cells Survive Matrix Disruption.

Biofilms are commonly recalcitrant to antibiotics, through incompletely elucidated mechanisms such as tolerance and persistence. We aimed at investigating how a Pseudomonas aeruginosa biofilm escapes ciprofloxacin treatment. P. aeruginosa PA14 in vitro mature biofilms were challenged with supra-MIC ciprofloxacin concentrations. Cell viability was quantified by fluorescein diacetate assay. Population dynamics were determined by counts of surviving culturable cells. Biofilms were analyzed using confocal laser scanning microscopy (CLSM), and the expression of genes involved in stringent response, toxin-antitoxin HigB/HigA, and type 3 secretion system (T3SS) was quantified by RT-qPCR in untreated and treated biofilms. Ciprofloxacin exposure resulted in an initial reduction of bacterial counts following a biphasic time-kill curve. After 24 h of treatment, the overall cell activity and the density of culturable cells significantly decreased as compared to untreated biofilm. No resistant mutant was isolated among the <1% surviving cells. Phenotypic adaptation toward persistence appeared to start after only 1 h of antibiotic exposure, by an overexpression of the genes involved in stringent response and in the toxin-antitoxin system, whereas the expression of genes encoding for the T3SS remained unchanged. After 4 h of ciprofloxacin exposure, stringent response genes returned to their basal level of expression. After a prolonged ciprofloxacin exposure, a deep alteration in the matrix structure that became thinner and lost mushroom-like aggregates was observed, in relation with reduced biovolumes of exopolysaccharides and extracellular DNA. These results support that ciprofloxacin might first induce the bacterial killing of most bacterial cells, but simultaneously activate stringent response mechanisms contributing to the switch of a subpopulation toward a persister phenotype. Once the persister phenotype is expressed, and despite an unexpected alteration of the biofilm matrix, ciprofloxacin fails to eradicate biofilm.

Soft sweep development of resistance in Escherichia coli under fluoroquinolone stress.

We employed a stepwise selection model for investigating the dynamics of antibiotic-resistant variants in Escherichia coli K-12 treated with increasing concentrations of ciprofloxacin (CIP). Firstly, we used Sanger sequencing to screen the variations in the fluoquinolone target genes, then, employed Illumina NGS sequencing for amplicons targeted regions with variations. The results demonstrated that variations G81C in gyrA and K276N and K277L in parC are standing resistance variations (SRVs), while S83A and S83L in gyrA and G78C in parC were emerging resistance variations (ERVs). The variants containing SRVs and/or ERVs were selected successively based on their sensitivities to CIP. Variant strain 1, containing substitution G81C in gyrA, was immediately selected following ciprofloxacin exposure, with obvious increases in the parC SRV, and parC and gyrA ERV allele frequencies. Variant strain 2, containing the SRVs, then dominated the population following a 20× increase in ciprofloxacin concentration, with other associated allele frequencies also elevated. Variant strains 3 and 4, containing ERVs in gyrA and parC, respectively, were then selected at 40× and 160× antibiotic concentrations. Two variants, strains 5 and 6, generated in the selection procedure, were lost because of higher fitness costs or a lower level of resistance compared with variants 3 and 4. For the second induction, all variations/indels were already present as SRVs and selected out step by step at different passages. Whatever the first induction or second induction, our results confirmed the soft selective sweep hypothesis and provided critical information for guiding clinical treatment of pathogens containing SRVs.

Antibiotic-Induced, Increased Conjugative Transfer Is Common to Diverse Naturally Occurring ESBL Plasmids in Escherichia coli.

Previously, we showed that cefotaxime (CTX) exposure increases conjugative transfer of a blaCTX–M–1 encoding IncI1 plasmid (IncI1/pST49/CTX-M-1) in Escherichia coli in a SOS-independent manner. This study aimed at investigating whether the observation was unique for that plasmid/strain/antibiotic combination or whether antibiotic-induced plasmid transfer (PT) is a more general phenomenon among plasmids in E. coli. Whole genome sequences of 25 E. coli strains were analyzed to identify different extended spectrum beta-lactamases (ESBL) plasmids enabling selection of a diverse collection of plasmids. Experiments were performed following exposure of these strains to 1/2 minimal inhibitory concentration (MIC) of CTX, ampicillin (AMP), or ciprofloxacin (CIP) before conjugation experiments. The frequency of PT was measured and compared to that of donors not exposed to antibiotics. Reverse-transcribed-quantitative real time polymerase chain reaction (RT-qPCR) was used to measure mRNA levels of five PT genes and two SOS response genes in donors exposed to antibiotics. The PT of eight strains (30.8% of strains tested) with IncI1/pST7/CTX-M-1, IncI1/pST49/CTX-M-1, IncI1/pST3/CTX-M-1, IncI1/pST293/CTX-M-1, IncI1/pST295/CTX-M-1, IncI1/pST16/CTX-M-55, and IncFII/CTX-M-14 (n = 2) plasmids was significantly increased following antibiotic exposure. CTX increased PT in all of these eight strain/plasmid combinations, AMP and CIP increased the PT in six and three strains, respectively. RT-qPCR showed that PT genes were up-regulated in the presence of the three antibiotics, whereas SOS-response genes were up-regulated only following CIP exposure. Our findings reveal that antibiotics can increase PT in E. coli strains with various ESBL plasmids. Thus, antibiotic-induced conjugative transfer of ESBL plasmids appears to be a common phenomenon in E. coli, having important implications for assessing the risks of antibiotic use.

Ecological impact of the antibiotic ciprofloxacin on microbial community of aerobic activated sludge.

This study investigated the effects and fate of the antibiotic ciprofloxacin (CIP) at environmentally relevant levels (50-500µg/L) in activated sludge (AS) microbial communities under aerobic conditions. Exposure to 500µg/L of CIP decreased species diversity by about 20% and significantly altered the phylogenetic structure of AS communities compared to those of control communities (no CIP exposure), while there were no significant changes upon exposure to 50µg/L of CIP. Analysis of community composition revealed that exposure to 500µg/L of CIP significantly reduced the relative abundance of Rhodobacteraceae and Nakamurellaceae by more than tenfold. These species frequently occur in AS communities across many full-scale wastewater treatment plants and are involved in key ecosystem functions (i.e., organic matter and nitrogen removal). Our analyses showed that 50-500µg/L CIP was poorly removed in AS (about 20% removal), implying that the majority of CIP from AS processes may be released with either their effluents or waste sludge. We therefore strongly recommend further research on CIP residuals and/or post-treatment processes (e.g., anaerobic digestion) for waste streams that may cause ecological risks in receiving water bodies.

Stress-responses of activated sludge and anaerobic sulfate-reducing bacteria sludge under long-term ciprofloxacin exposure

The activated sludge (AS) and sulfate-reducing bacteria (SRB) sludge systems were continuously operated for 200 days in laboratory to investigate the stress-responses of these two sludge systems under ciprofloxacin (CIP) exposure. It was found that CIP was effectively removed by SRB sludge via adsorption and biodegradation, but little biodegradation in AS system. The CIP biodegradation by SRB sludge made the SRB sludge system more sustainable and tolerant to long-term CIP exposure than AS system with significant (p < 0.05) CIP desorption and decrease of CIP removal. CIP shaped the microbial communities in AS and SRB sludge, and significantly (p < 0.05) inhibited the family Nitrosomonadaceae (ammonia-oxidizing bacteria (AOB)) and genus Nitrospira (nitrite-oxidizing bacteria (NOB)/complete ammonia oxidizer(comammox)) and the nitrogen removal in AS system. Moreover, CIP posed the increase of genus Zoogloea-like organisms and the non-filamentous bulking of AS, e.g. 313 ± 12 mL/g of sludge volume index (SVI) at phase V (influent CIP = 5000 μg/L). The genus Desulfobacter was enriched in SRB sludge system under long-term CIP exposure, and stimulated chemical oxygen demand (COD) removal and sulfate reduction. The increase of genera Zoogloea, Acinetobacter and Flavobacterium in AS, and Zoogloea and Acinetobacter in SRB sludge systems under CIP exposure promoted extracellular polymeric substances (EPS) production and CIP adsorption for self-protection of microbes against CIP toxicity. The functional groups of N–H, O–H, C–O–C and C=O in EPS of AS and SRB sludge provided adsorption sites for CIP and impeded CIP impact on microbial cells. The findings of this study provide an insight into the stress-responses of AS and SRB sludge under long-term CIP exposure, and exhibit the great potential of treating CIP-laden wastewater by SRB sludge system.

Toxicological effects of ciprofloxacin exposure to Drosophila melanogaster

The abuse of ciprofloxacin (CIP) may cause serious side effects and the mechanisms underlying these effects remain unclear. Here, we determinate the 48 h, 72 h and 96 h LC50 values of CIP to Drosophila melanogaster and demonstrate a series of adverse effects after D. melanogaster was exposed to CIP at a sublethal concentration (3.2 mg mL−1). Treated individuals showed shorter lifespan, delayed development and many of the treated larvae failed to pupate or hatch. Smaller body size was observed at every life stage when exposed to CIP and the size of pupae, the weight of third-instar larvae exhibited a perfectly dose-response relationship that the larger concentration exposed to, the smaller body size or lighter weight is. Moreover, reduction in fat body cell viability, elevated oxidative stress markers (SOD and CAT) and down-regulation of diap1, ex, two target genes of Yorkie (Yki), was observed in response to CIP exposure. Most importantly, we found two types of black spot in Drosophila and the proportion of larvae with a black spot was positively related to the treatment dose, which is new in the field. This study provides a scientific basis for the potential harm caused by abuse of quinolones with the goal of urging cautious use of antibiotics.

The cardiovascular toxicity induced by high doses of gatifloxacin and ciprofloxacin in zebrafish

As a new type of pollutant, fluoroquinolones (FQs) antibiotics are ubiquitous in environment and have some threat to human health and ecological environment. Their ecological toxicity to the environment urgently need to be assessed. Therefore, we firstly explored the toxic effects and possible mechanism of cardiovascular toxicity induced by gatifloxacin (GTFX) and ciprofloxacin (CPFX) using zebrafish model. After 24 h exposure, the zebrafish treated with GTFX showed pericardial edema which was further investigated by histopathological examination, while CPFX exposure did not induce morphological abnormalities. However, both of them induced cardiac dysfunction, such as decreased heart rate and cardiac output which was showed a positive correlation with the concentration. To better understand the possible molecular mechanisms underlying cardiovascular toxicity in zebrafish, we investigated the transcriptional level of genes related to calcium signaling pathway and cardiac muscle contraction. The results indicated that the expression of ATPase (atp2a1l) and cardiac troponin C (tnnc1a) genes were significantly inhibited, the expression of calcium channel (cacna1ab) gene showed slight promoted trend after CPFX exposure. For zebrafish treated with GTFX, the expression of atp2a1l genes was also significantly inhibited, while the expression of tnnc1a genes was slightly inhibited and cacna1ab genes expression had no obvious effect. The present study firstly revealed that GTFX exposure can induce morphological and functional abnormalities on the cardiovascular system of zebrafish. Though CPFX exposure did not induce morphological abnormalities, the function of cardiovascular system was still damaged. Mechanistically, this toxicity might result from the pressure of down-regulation of genes associated with calcium signaling pathway and cardiac muscle contraction. The results of this study can provide a valuable theoretical basis for the establishment of FQs environmental quality standards in water environment, environmental drug regulation and risk management.

Salmonella enterica persister cells form unstable small colony variants after in vitro exposure to ciprofloxacin

Persistence phenotype and small colony variants (SCVs) can be part of a bacterial bet-hedging strategy for survival under environmental stresses, such as antimicrobial exposure. These phenotypes are of particular concern in persistent and relapsing infections, since cells resume to normal growth after cessation of the stressful condition. In this context, we found persisters and unstable SCVs as phenotypic variants of Salmonella enterica that were able to survive ciprofloxacin exposure. A high heterogeneity in persister levels was observed among S. enterica isolates grown under planktonic and biofilm conditions and exposed to ciprofloxacin or ceftazidime, which may indicate persistence as a non-multidrug-tolerant phenotype. Nevertheless, a comparable variability was not found in the formation of SCVs among the isolates. Indeed, similar proportions of SCV in relation to normal colony phenotype (NCP) were maintained even after three successive cycles of ciprofloxacin exposure testing colonies from both origins (SCV or NCP). Additionally, we found filamentous and dividing cells in the same scanning electron microscopy images from both SCV and NCP. These findings lead us to hypothesize that besides variability among isolates, a single isolate may generate distinct populations of persisters, where cells growing under distinct conditions may adopt different and perhaps complementary survival strategies.

Pharmacodynamics of ciprofloxacin against Pseudomonas aeruginosa planktonic and biofilm-derived cells.

The influence of growth phase and state on the survival and recovery of Pseudomonas aeruginosa exposed to ciprofloxacin was investigated using batch culture grown planktonic cells and disaggregated biofilm populations. Biofilms were either nonantibiotic exposed or previously exposed to ciprofloxacin before disaggregation and subsequent challenge with ciprofloxacin. Viable counts showed that late stationary phase cells were tolerant to ciprofloxacin over 24h exposure, while all other populations presented a biphasic killing pattern. In contrast, the metabolic activity of planktonic and biofilm-derived cells remained similar to controls during the initial 6h of ciprofloxacin exposure, despite a significant reduction in viable cell numbers. A similar effect was observed when assessing the postantibiotic effect of 1h ciprofloxacin exposure. Thus, although cell reduction occurred, the metabolic status of the cells remained unchanged. The recovery of disaggregated biofilm cells previously exposed to ciprofloxacin was significantly quicker than naïve biofilm cells, and this latter population's recovery was significantly slower than all planktonic populations. Results from this work have implications for our understanding of biofilm-related infections and their resilience to antimicrobial treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: Removal of biofilms from surfaces and infection sites via disaggregation and induction of dispersion may reverse their antibiotic tolerant state. However, little is known of the recovery of the cells upon disaggregation from biofilms. Driven by this gap in knowledge we quantified the effect of ciprofloxacin on disaggregated biofilms of Pseudomonas aeruginosa, including those previously exposed to ciprofloxacin. Our results provide further insight into bacterial resilience, regrowth, and antimicrobial efficacy, as reduction in cell viability does not directly correlate with the metabolic activity of bacteria at the time of the exposure to antimicrobials. Thus, despite a perceived reduction in viability, the potential for cell persistence and regrowth remains and recovery is quicker upon subsequent exposure to antimicrobial, supporting the increase in resilience and recurrence of infections.

Evaluation of the taxonomic and functional variation of freshwater plankton communities induced by trace amounts of the antibiotic ciprofloxacin

Ciprofloxacin (CIP), one of the most frequently detected antibiotics in water systems, has become an aquatic contaminant because of improper disposal and excretion by humans and animals. It is still unknown how trace amounts of CIP affect the aquatic microbial community diversity and function. We therefore investigated the effects of CIP on the structure and function of freshwater microbial communities via 16S/18S rRNA gene sequencing and metatranscriptomic analyses. CIP treatment (7 μg/L) did not significantly alter the physical and chemical condition of the water body as well as the composition of the main species in the community, but slightly increased the relative abundance of cyanobacteria and decreased the relative abundance of eukaryotes. Metatranscriptomic results showed that bacteria enhanced their phosphorus transport and photosynthesis after CIP exposure. The replication, transcription, translation and cell proliferation were all suppressed in eukaryotes, while the bacteria were not affected in any of these aspects. This interesting phenomenon was the exact opposite to both the antibacterial property of CIP and its safety for eukaryotes. We hypothesize that reciprocal and antagonistic interactions in the microcosm both contribute to this result: cyanobacteria may enhance their tolerance to CIP through benefiting from cross-feeding and some secreted substances that withstand bacterial CIP stress would also affect eukaryotic growth. The present study thus indicates that a detailed assessment of the aquatic ecotoxicity of CIP is essential, as the effects of CIP are much more complicated in microbial communities than in monocultures. CIP will continue to be an environmental contaminant due to its wide usage and production and more attention should be given to the negative effects of antibiotics as well as other bioactive pollutants on aquatic environments.