Presence Of Fluoroquinolones Research Articles

A triple-channel sensor array utilizing fluorescent carbon dots for simultaneous discrimination and detection of multiple fluoroquinolones

The presence of fluoroquinolones (FQs) residues in food and the environment has prompted concerns regarding food safety and public health. Consequently, it is of great significance to analyze the types and levels of FQs present. However, the majority of studies have concentrated on the specific detection of individual FQs, with a notable absence of high-throughput and rapid analysis methods for the simultaneous detection of multiple FQs that may coexist in food and the environment. Hereon, a triple-channel sensor array was successfully constructed utilizing fluorescent carbon dots (TA-CDs), with the assistance of Cu2+ and Fe3+, for the qualitative discrimination and quantitative detection of eight types of FQs. The sensor array can distinguish between different concentrations of FQs and various mixtures of FQs, as well as 100 % accuracy in the discrimination of unknown samples. Impressively, the sensor platform can quantitatively detect FQs in animal-derived foods, such as honey, milk, eggs, and pork, as well as in water samples. This research has the potential to be extended to other analytes with similar chemical structures or properties.

Rapid, DNA-induced interface swapping by DNA gyrase

DNA gyrase, a ubiquitous bacterial enzyme, is a type IIA topoisomerase formed by heterotetramerisation of 2 GyrA subunits and 2 GyrB subunits, to form the active complex. DNA gyrase can loop DNA around the C-terminal domains (CTDs) of GyrA and pass one DNA duplex through a transient double-strand break (DSB) established in another duplex. This results in the conversion from a positive (+1) to a negative (–1) supercoil, thereby introducing negative supercoiling into the bacterial genome by steps of 2, an activity essential for DNA replication and transcription. The strong protein interface in the GyrA dimer must be broken to allow passage of the transported DNA segment and it is generally assumed that the interface is usually stable and only opens when DNA is transported, to prevent the introduction of deleterious DSBs in the genome. In this paper, we show that DNA gyrase can exchange its DNA-cleaving interfaces between two active heterotetramers. This so-called interface ‘swapping’ (IS) can occur within a few minutes in solution. We also show that bending of DNA by gyrase is essential for cleavage but not for DNA binding per se and favors IS. Interface swapping is also favored by DNA wrapping and an excess of GyrB. We suggest that proximity, promoted by GyrB oligomerization and binding and wrapping along a length of DNA, between two heterotetramers favors rapid interface swapping. This swapping does not require ATP, occurs in the presence of fluoroquinolones, and raises the possibility of non-homologous recombination solely through gyrase activity. The ability of gyrase to undergo interface swapping explains how gyrase heterodimers, containing a single active-site tyrosine, can carry out double-strand passage reactions and therefore suggests an alternative explanation to the recently proposed ‘swivelling’ mechanism for DNA gyrase (Gubaev et al., 2016).

Rapid, DNA-induced interface swapping by DNA gyrase.

DNA gyrase, a ubiquitous bacterial enzyme, is a type IIA topoisomerase formed by heterotetramerisation of 2 GyrA subunits and 2 GyrB subunits, to form the active complex. DNA gyrase can loop DNA around the C-terminal domains (CTDs) of GyrA and pass one DNA duplex through a transient double-strand break (DSB) established in another duplex. This results in the conversion from a positive (+1) to a negative (-1) supercoil, thereby introducing negative supercoiling into the bacterial genome by steps of 2, an activity essential for DNA replication and transcription. The strong protein interface in the GyrA dimer must be broken to allow passage of the transported DNA segment and it is generally assumed that the interface is usually stable and only opens when DNA is transported, to prevent the introduction of deleterious DSBs in the genome. In this paper, we show that DNA gyrase can exchange its DNA-cleaving interfaces between two active heterotetramers. This so-called interface 'swapping' (IS) can occur within a few minutes in solution. We also show that bending of DNA by gyrase is essential for cleavage but not for DNA binding per se and favors IS. Interface swapping is also favored by DNA wrapping and an excess of GyrB. We suggest that proximity, promoted by GyrB oligomerization and binding and wrapping along a length of DNA, between two heterotetramers favors rapid interface swapping. This swapping does not require ATP, occurs in the presence of fluoroquinolones, and raises the possibility of non-homologous recombination solely through gyrase activity. The ability of gyrase to undergo interface swapping explains how gyrase heterodimers, containing a single active-site tyrosine, can carry out double-strand passage reactions and therefore suggests an alternative explanation to the recently proposed 'swivelling' mechanism for DNA gyrase (Gubaev et al., 2016).

Fluorescent responsive membrane based on terbium coordination polymer and carbon dots with AIE effect for rapid and visual detection of fluoroquinolone

In this study, based on aggregation-induced emission (AIE) effect and antenna effect, a novel portable fluorescent responsive membrane was constructed with red carbon dots (R-CDs) as reference signal and terbium coordination polymer (Tb-AMP CPs) as response signal for visual, instrument-free, and sensitive detection of fluoroquinolones (FQs). Specifically, the fluorescent responsive membrane (R-T membrane) was prepared by physically depositing R-CDs with AIE property and Tb-AMP CPs on the surface of polyvinylidene fluoride filter membranes at ambient temperature. In the presence of FQs, Tb3+ in the Tb-AMP CPs of the prepared membrane coordinated with the β-diketone structure of FQs, which turned on the yellow-green fluorescence through the “antenna effect”. As the concentration of FQs increased, the R-T membrane achieved a fluorescent color transition from bright pink to yellow-green. Its visual detection sensitivity for three FQs, including ciprofloxacin, difloxacin, and enrofloxacin, was 0.01 μM, and the detection limits were 7.4 nM, 7.8 nM, and 9.2 nM, respectively, by analyzing the color parameter green. In the residue analysis of FQs in real samples, the constructed membrane also exhibited remarkable anti-interference and reliability, which is of great significance for ensuring the safety of animal-derived food.

Integrating target-responsive microfluidic-based biosensing chip with smartphone for simultaneous quantification of multiple fluoroquinolones

The presence of fluoroquinolone (FQs) antibiotic residues in the food and environment has become a significant concern for human health and ecosystems. In this study, the background-free properties of upconversion nanoparticles (UCNPs), the high specificity of the target aptamer (Apt), and the high quenching properties of graphene oxide (GO) were integrated into a microfluidic-based fluorescence biosensing chip (MFBC). Interestingly, the microfluidic channels of the MFBC were prepared by laser-printing technology without the need for complex preparation processes and additional specialized equipment. The target-responsive fluorescence biosensing probes loaded on the MFBC were prepared by self-assembly of the UCNPs-Apt complex with GO based on π-π stacking interactions, which can be used for the detection of the two FQs on a large scale without the need for multi-step manipulations and reactions, resulting in excellent multiplexed, automated and simultaneous sensing capabilities with detection limits as low as 1.84 ng/mL (enrofloxacin) and 2.22 ng/mL (ciprofloxacin). In addition, the MFBC was integrated with a smartphone into a portable device to enable the analysis of a wide range of FQs in the field. This research provides a simple-to-prepare biosensing chip with great potential for field applications and large-scale screening of FQs residues in the food and environment.

An innovative probe based on aluminum ion-triggered aggregation induced emission effect for ratiometric fluorescence visual detection of fluoroquinolone

Based on the aggregation-induced emission effect triggered by Al3+, a novel probe was constructed for the ratiometric fluorescence visualization detection of fluoroquinolones (FQs) residues. In this probe, Al3+ cased the glutathione-modified copper nanoclusters (GSH-CuNCs) to aggregate by the interaction with surface carboxyl groups, restricting the intramolecular vibration and rotation of GSH-CuNCs and reducing the non-radiative relaxation of excited states. Significant enhancement of fluorescence and quantum yield were achieved. In the presence of FQs, Al3+/GSH-CuNCs coordinated with FQs to form a polymer that inhibited intramolecular proton transfer in the excited state of FQs, thereby enhancing the intrinsic fluorescence of FQs, while the fluorescence intensity of Al3+/GSH-CuNCs was not disturbed. The probe solution changed from red to blue (or green) under UV lamp as the concentration of FQs increased, providing the possibility for semi-quantitative visual analysis. It also showed sensitive spectral responses to FQs with detection limits of 7.9 (ciprofloxacin), 6.8 (gatifloxacin), and 9.5 nM (enrofloxacin), respectively. The application potential in the detection of FQs residues in poultry eggs was verified by real sample test. Additionally, the probe has been extended to the development of fluorescent test paper and the smartphone detection platform, contributing to rapid, portable, visual quantitative analysis of FQs.

Unveil the mechanism of photosensitized fluoroquinolones enhancing chlortetracycline photodegradation under simulated sunlight: Batch experiments and DFT calculation

Fluoroquinolones (FQs), as the most commonly used antibiotics, are ubiquitous in the aquatic environment. The FQs' self-sensitization process could generate reactive oxygen species (ROS), which could react with other coexisting organic pollutants, impacting their transformation behaviors. However, the FQs’ influences and mechanisms on the photochemical transformation of coexisting antibiotics are not yet revealed. In this study, we found ofloxacin (OFL) and norfloxacin (NOR), the two common FQs, can obviously accelerate chlortetracycline (CTC) photodegradation. In the presence of OFL and NOR (i.e., 10 μM), CTC photodegradation rate constants increased by 181.1% and 82.9%, respectively. With the help of electron paramagnetic resonance (EPR) and quenching experiments, this enhancement was attributed to aromatic ketone structure in FQs, which absorbed photons to generate ROS (i.e., 3OFL*, 3NOR*,1O2, and •OH). Notably, 3OFL* or 3NOR* was dominantly contributed to the enhanced CTC photodegradation, with the contribution ratios of 79.9% and 77.3% in CTC indirect photodegradation, respectively. Compared to CTC direct photodegradation, some new photodegradation products were detected in FQs solution, suggesting that 3OFL* or 3NOR* may oxide CTC through electron transfer. Moreover, the higher triple-excited state energy of OFL and NOR over DFT calculation implied that energy transfer from 3OFL* or 3NOR* to CTC was also theoretically feasible. Therefore, the presence of FQs could significantly accelerate the photodegradation of coexisting antibiotics mainly via electron or energy transfer of 3FQs*. The present study provided a new insight for accurately evaluating environmental behaviors and risks when multiple antibiotics coexist.

Host Cell Oxidative Stress Promotes Intracellular Fluoroquinolone Persisters of Streptococcus pneumoniae.

Bacterial persisters represent a small subpopulation that tolerates high antibiotic concentrations without acquiring heritable resistance, and it may be generated by environmental factors. Here, we report the first antibiotic persistence mechanism in Streptococcus pneumoniae, which is induced by oxidative stress conditions and allows the pneumococcus to survive in the presence of fluoroquinolones. We demonstrated that fluoroquinolone persistence is prompted by both the impact of growth arrest and the oxidative stress response induced by H2O2 in bacterial cells. This process protected pneumococci against the deleterious effects of high ROS levels induced by fluoroquinolones. Importantly, S. pneumoniae develops persistence during infection, and is dependent on the oxidative stress status of the host cells, indicating that its transient intracellular life contributes to this mechanism. Furthermore, our findings suggest persistence may influence the outcome of antibiotic therapy and be part of a multistep mechanism in the evolution of fluoroquinolone resistance. IMPORTANCE In S. pneumoniae, different mechanisms that counteract antibiotic effects have been described, such as vancomycin tolerance, heteroresistance to penicillin and fluoroquinolone resistance, which critically affect the therapeutic efficacy. Antibiotic persistence is a type of antibiotic tolerance that allows a bacterial subpopulation to survive lethal antimicrobial concentrations. In this work, we used a host-cell infection model to reveal fluoroquinolone persistence in S. pneumoniae. This mechanism is induced by oxidative stress that the pneumococcus must overcome to survive in host cells. Many fluoroquinolones, such as levofloxacin and moxifloxacin, have a broad spectrum of activity against bacterial pathogens of community-acquired pneumonia, and they are used to treat pneumococcal diseases. However, the emergence of fluoroquinolone-resistant strains complicates antibiotic treatment of invasive infections. Consequently, antibiotic persistence in S. pneumoniae is clinically relevant due to prolonged exposure to fluoroquinolones likely favors the acquisition of mutations that generate antibiotic resistance in persisters. In addition, this work contributes to the knowledge of antibiotic persistence mechanisms in bacteria.

Preparation of green sodium alginate adsorption membrane and its high adsorption performance for fluoroquinolones antibiotics

Fluoroquinolones (FQs) were widely used in human and veterinary medicine, however, the presence of FQs at lower concentration in the environment could threaten to human health and agricultural production. In this research, the adsorption membranes were prepared based on sodium alginate (SA) crosslinking with multivalent metal ions. The SA-Al/Ca membranes and SA-Ca membrane were prepared by crosslinking with AlCl3/CaCl2 solutions and CaCl2 solutions, respectively. Comparing with SA-Ca membrane, the presence of Al3+ in crosslinking agent could produce a large number of adsorption sites on the surface of SA-Al/Ca membrane and remarkably improve the adsorption performance of SA-Al/Ca membrane. Effects of preparation conditions such as crosslinking ways, CaCl2 percentage in compound crosslinker, and crosslinking time on the adsorption performance of prepared SA-Al/Ca membrane were demonstrated. Moreover, the effects of the initial concentration of ciprofloxacin (CIP) and Levofloxacin (S-OFX), adsorption time, solution pH, ionic strength and adsorption temperature on the performance of SA-Al/Ca membrane were studied. The thermodynamics of the adsorption of CIP and S-OFX were well fitted with the Tempkin adsorption isotherm model (R2 = 0.9455) and D-R adsorption isotherm model (R2 = 0.9822), respectively. The maximum adsorption capacity of S-OFX and CIP using SA-Al/Ca membrane was 836.31 mg·g−1 and 858.94 mg·g−1. The results showed that SA-Al/Ca membranes with good adsorption capacity, low cost and high stability.

Fluoroquinolone antibiotics sensitized photodegradation of isoproturon

Fluoroquinolone (FQ) antibiotics are a group of contaminants of emerging environmental concern. In the present study, we demonstrated that norfloxacin (NORF) and ofloxacin (OFLO), two typical FQs, have photochemical reactivity analogous to chromophoric dissolved natural organic matter (DOM) in surface waters and can sensitize the photodegradation of isoproturon (IPU), a phenylurea herbicide. Such photochemical reactivity is ascribed to the quinolone chromophore that is excited to a triplet state (3FQ*) upon UV-A irradiation. 3FQ* further reacts with dissolved oxygen to give rise to singlet oxygen. 3FQ* steady-state concentrations of 6.72 × 10−15 and 1.27 × 10−15 M were measured in 10 μM NORF and OFLO solutions, respectively, under UV365nm irradiation. The degradation of IPU was due to the reaction with 3FQ*, with bimolecular rate constants of 6.07 × 109 and 1.51 × 1010 for 3NORF* and 3OFLO*, respectively. Intriguingly, NORF and OFLO per se were unstable and photolyzed during UV-A irradiation, but the photochemical reactivities of the solutions were not lost accordingly. High-resolution mass spectrometry analysis revealed that defluorination and piperazine moiety oxidation were the main photolysis pathways, while the core quinolone structure remained intact. Thus, the photolysis products largely inherited the photochemical reactivity of the parent compounds. Since all FQs share the same quinolone structure, similar photochemical reactivity is expected. The presence of FQs in surface water would affect the transformation and fate of coexisting compounds. To the best of our knowledge, this is the first study examining the environmental behavior of FQs as photosensitizers. The findings greatly advance the understandings of the influence of FQs in aquatic environment.

Shifts of bacterial community and molecular ecological network at the presence of fluoroquinolones in a constructed wetland system

Fluoroquinolones (FQs) has caused increasing concerns regarding its potential environmental risks. However, their effects on bacterial community and microbial interactions in wetland system remains unclear. To verify these issues, a lab-scale constructed wetland exposed to low concentration FQs mixture was carried out for two months. The results showed that the removal efficiencies of COD and TP were negatively affected. FQs significantly increased the bacterial diversity and altered the overall bacterial community structure. Proteobacteria significantly decreased while Firmicutes exhibited opposite tendency (P < 0.05). Dechloromonas and Delftia, involved in phosphorus removal, decreased significantly (P < 0.05). Molecular ecological network analysis suggested that FQs promoted the network complexity and microbial interactions. A super module emerged at FQs and among-module connections were weakened obviously. Additionally, Nodes of Betaproteobacteria lost most interactions while Clostridia acquired more interactions at the presence of FQs. This study provided insights into how the bacterial community and their molecular ecological network respond to FQs in constructed wetland system.

1144. Modern Problem, Medieval Cure-Resistant Aeromonas in Medicinal Leeches

BackgroundMedicinal leeches are used primarily in plastic and reconstructive surgery when venous congestion threatens tissue viability. The associated infection risk ranges from 4.1 to 20%. Prophylactic antimicrobials such as fluoroquinolones (FQ) or trimethoprim-sulfamethoxazole (SXT) are recommended and target commonly isolated pathogen and gut symbiont, Aeromonas. However, resistance to these agents has been reported and detected in leeches, including at hospital systems across Canada that acquire their stock from the same supplier. Our objective was to describe the local epidemiology of leech-related Aeromonas resistant to one or more commonly used prophylactic agents, and determine if such resistance originates from the common supplier.MethodsSix hospital systems across Canada using leech therapy, purchased from the same supplier, were surveyed. A 5-year retrospective review of all antimicrobial resistant leech-related Aeromonas, derived from clinical, leech, and tank fluid specimens was performed. All Aeromonas resistant to either FQ or SXT were included, and retained frozen isolates from each system were analysed by pulse-field gel electrophoresis (PFGE) using a published Aeromonas protocol.ResultsAll six hospital systems reported leech-related Aeromonas resistant to one or more antimicrobials, totalling 15 isolates. Three systems only reported data from the last year. Four systems used FQ and two used SXT as prophylaxis. Fifteen of 15 were either FQ resistant or intermediate, and four of 15 were SXT resistant. Three of 10 isolates tested for ceftriaxone (CRO) susceptibility were resistant. Five of 15 of the isolates were resistant to two or more agents. Of the two leech quality control isolates, 2/2 were FQ resistant and 1/2 was FQ, SXT and CRO resistant. Only three isolates, each from a different, geographically distinct hospital system, had been retained. PFGE analysis indicated 2/3 are closely related (Figure 1).ConclusionOur preliminary investigation suggests that the presence of FQ and SXT resistance in leech-related Aeromonas might be more common than previously suspected, and that such resistance might originate from a common source. A broader study of the molecular epidemiology of leech-related Aeromonas is warranted. Disclosures All authors: No reported disclosures.

ONE HEALTH: ENVIRONMENTAL CONTAMINATION BY ANTIMICROBIALS

Over the past few decades, a high number of pharmaceuticals have been detected in surface, ground and drinking waters. This contamination comes from domestic sewage, livestock, hospitals and chemical- pharmaceutical industries. Typical examples of these pollutants are the antimicrobials – powerful antibiotics used in human and veterinary medicine. The presence of antimicrobials in the environment can pose a serious threat to the ecosystem and to human health due to their high consumption globally. Even at low environmental concentrations, antibiotics stimulate bacterial resistance. The consequences of the presence of fluoroquinolones in the environment are not fully understood, but are known to be toxic to plants and aquatic organisms. Approximately 85% of the antimicrobials present in influents can be removed by conventional wastewater treatment plants, but the removed fraction is frequently accumulated in the sludge, which is sometimes used as fertilizer, representing an additional input route into the environment. The removal of antimicrobials by biological treatment is ineffective, and it is believed that only advanced oxidation technologies are able to destroy these emerging pollutants.

The effect of a commercial competitive exclusion product on the selection of enrofloxacin resistance in commensal E. coli in broilers

ABSTRACTThe effect of a competitive exclusion product (Aviguard®) on the selection of fluoroquinolone resistance in poultry was assessed in vivo in the absence or presence of fluoroquinolone treatment.Two experiments using a controlled seeder-sentinel animal model (2 seeders: 4 sentinels per group) with one-day-old chicks were used. For both experiments, as soon as the chicks were hatched, the birds of two groups were administered Aviguard® and two groups were left untreated. Three days later, all groups were inoculated with an enrofloxacin-susceptible commensal E. coli strain. Five days after hatching, two birds per group were inoculated with either a bacteriologically fit or a bacteriologically non-fit enrofloxacin-resistant commensal E. coli strain. In experiment 2, all groups were orally treated for three consecutive days (days 8–10) with enrofloxacin. Throughout the experiments, faecal excretion of all inoculated E. coli strains was determined on days 2, 5, 8, 11, 18 and 23 by selective plating (via spiral plater). Linear mixed models were used to assess the effect of Aviguard® on the selection of fluoroquinolone resistance.The use of Aviguard® (P < 0.01) reduced the excretion of enrofloxacin-resistant E. coli when no enrofloxacin treatment was administered. However, this beneficial effect disappeared (P = 0.37) when the birds were treated with enrofloxacin. Similarly, bacterial fitness of the enrofloxacin-resistant E. coli strain used for inoculation had an effect (P < 0.01) on the selection of enrofloxacin resistance when no treatment was administered, whereas this effect was no longer present when enrofloxacin was administered (P = 0.70).Thus, enrofloxacin treatment cancelled the beneficial effects from administrating Aviguard® in one-day-old broiler chicks and resulted in an enrofloxacin-resistant flora.RESEARCH HIGHLIGHTSThe effect of Aviguard® on the selection of enrofloxacin resistance was assessed in vivo.Without enrofloxacin, Aviguard® reduced the selection of enrofloxacin resistance.When enrofloxacin was administered, it cancelled the beneficial effect of Aviguard®.

In-vitro drug–drug interaction studies of diltiazem with floroquinolones

ABSTRACTDiltiazem is an established cardiovascular drug mainly used for the management of hypertension specifically for the angina pectoris. Fluoroquinolones are widely prescribed against the treatment of severe infections. In vitro relations of diltiazem with fluoroquinolones (ciprofloxacin, levofloxacin, norfloxacin, and ofloxacin) were examined using spectrophotometric and separation techniques, i.e., RP-HPLC. Diltiazem’s availabilities were observed to be predisposed highly in the presence of fluoroquinolones. To investigate the mechanism of interaction in a variety of dissolution environments, i.e., simulating body environments with regard to pH on these interactions has been studied. Moreover, complex of diltiazem–fluoroquinolones were prepared and elucidated through IR spectroscopy and confirmed by computational molecular modeling.

Efficient Reduction of Antibacterial Activity and Cytotoxicity of Fluoroquinolones by Fungal-Mediated N-Oxidation.

Extensive usage of fluoroquinolone antibiotics in livestock results in their occurrence in manure and subsequently in the environment. Fluoroquinolone residues may promote bacterial resistance and are toxic to plants and aquatic organisms. Moreover, fluoroquinolones may enter the food chain through plant uptake, if manure is applied as fertilizer. Thus, the presence of fluoroquinolones in the environment may pose a threat to human and ecological health. In this study, the biotransformation of enrofloxacin, marbofloxacin, and difloxacin by the fungus X. longipes (Xylaria) was investigated. The main metabolites were unequivocally identified as the respective N-oxides by mass spectrometry and nuclear magnetic resonance spectroscopy. Fungal-mediated N-oxidation of fluoroquinolones led to a 77-90% reduction of the initial antibacterial activity. In contrast to their respective parent compounds, N-oxides showed low cytotoxic potential and had a reduced impact on cell proliferation. Thus, biotransformation by X. longipes may represent an effective method for inactivating fluoroquinolones.

Effects of Fluoroquinolones and Azithromycin on Biofilm Formation of Stenotrophomonas maltophilia.

Stenotrophomonas maltophilia is an opportunistic pathogen that causes respiratory and urinary tract infections, as well as wound infections in immunocompromised patients. This pathogen is difficult to treat due to increased resistance to many antimicrobial agents. We investigated the in vitro biofilm formation of S. maltophilia, including effects of fluoroquinolones (FQs) and azithromycin on biofilm formation. The organism initiated attachment to polystyrene surfaces after a 4 h incubation period, and reached maximal growth at 18–24 h. In the presence of FQs (moxifloxacin, levofloxacin or ciprofloxacin), the biofilm biomass was significantly reduced (P < 0.05). A lower concentration of moxifloxacin (10 μg/mL) exhibited a better inhibiting effect on biofilm formation than 100 μg/mL (P < 0.01), but with no difference in effect compared to the 50 μg/mL concentration (P > 0.05). However, the inhibitory effects of 10 μg/mL of levofloxacin or ciprofloxacin were slightly less pronounced than those of the higher concentrations. A combination of azithromycin and FQs significantly reduced the biofilm inhibiting effect on S. maltophilia preformed biofilms compared to azithromycin or FQs alone. We conclude that early use of clinically acceptable concentrations of FQs, especially moxifloxacin (10 μg/mL), may possibly inhibit biofilm formation by S. maltophilia. Our study provides an experimental basis for a possible optimal treatment strategy for S. maltophilia biofilm-related infections.

Fluoroquinolone pollution of food, water and soil, and bacterial resistance

Fluoroquinolones are a valuable synthetic antibacterial class widely used in the treatment of infectious diseases both in humans and animals. Until recently, it has been thought that bacterial resistance to fluoroquinolones develops very slowly. Nowadays, there are multiple studies that reveal the alarming occurrence of bacterial resistance and there is a high risk of becoming therapeutically useless. The emergence of this phenomenon comes from injudicious usage in therapy, the presence of residues and their metabolites in food of animal origin and also in sewage, compost and domestic waste, which end up in soil and water sources. In the present paper, we reviewed important issues regarding fluoroquinolones impact on the environment in connection with the development of bacterial resistance: (1) the presence of fluoroquinolones as pollutants in soil, surface waters, and food. Fluoroquinolones are persistent with high specificity to interact with soil compared to other antibiotics. Pollution of water sources raises concerns regarding the effects of small concentrations (ng L−1) on human health and also of the environment. The non-therapeutic use in animal farms conducts to food pollution; the cultivated plants could concentrate the fluoroquinolones (over 100 μg L−1); (2) the increase of bacterial resistance to fluoroquinolones occurring with specific mutations in the target enzymes as well by the plasmid-mediated resistance and active efflux of the cell; (3) international regulations of the fluoroquinolone residues in food that are far to encompass all compounds; (4) fluoroquinolones residues analysis with standardized methods should provide limits of detection lower than maximum residue limit values; and (5) trends and perspectives: (a) a wider process of harmonization of regulations; (b) the fluoroquinolones restriction, necessary for low levels of bacterial resistance; (c) the soil and waste water purification methods; (d) the practice of soil planting scheme as an alternative; and (e) an environmental label in order to facilitate the selection of drugs.

Effect of land application of manure from enrofloxacin-treated chickens on ciprofloxacin resistance of Enterobacteriaceae in soil

A field plot experiment was carried out to evaluate the impact of spreading chicken manure containing enrofloxacin (ENR) and its metabolite ciprofloxacin (CIP), on the levels of CIP-resistant Enterobacteriaceae in soil. The manures from chickens treated with ENR and from untreated control chickens were applied on six plots. Total and CIP-resistant Enterobacteriaceae were counted on Violet Red Bile Glucose medium containing 0 to 16mgL−1 of CIP. A total of 145 isolates were genotyped by enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR). The minimum inhibitory concentration (MIC) of CIP for the isolates of each ERIC-PCR profile was determined. The most frequently isolated Enterobacteriaceae included Escherichia coli, and to a lesser extent, Enterobacter and 5 other genera from environmental origin. The composition of the E. coli community differed between manure and manured soil suggesting that the E. coli genotypes determined by ERIC-PCR varied significantly in their ability to survive in soil. One of these genotypes, including both susceptible and resistant isolates, was detected up to 89days after the manure was applied. Most of the E. coli isolated in soil amended with manure from treated chickens was resistant to CIP (with a MIC ranging between 2 and 32mgL−1). In contrast, despite the presence of ENR in soil at concentrations ranging from 13–518μgkg−1, the environmental Enterobacteriaceae isolates had a CIP MIC≤0.064mgL−1, except one isolate which had a MIC of 0.25mgL−1, These results showed that spreading manure from ENR-treated chickens enabled CIP-resistant E. coli to persist for at least three months in the soil. However, neither the presence of fluoroquinolones, nor the persistence of CIP-resistant E. coli, increased the CIP-susceptibility of environmental Enterobacteriaceae.

Performance of aerobic granular sludge in a sequencing batch bioreactor exposed to ofloxacin, norfloxacin and ciprofloxacin

A granular sludge sequencing batch reactor (SBR) was operated for 340 days for treating a synthetic wastewater containing fluoroquinolones (FQs), namely ofloxacin, norfloxacin and ciprofloxacin. The SBR was intermittently fed with FQs, at concentrations of 9 and 32 μM. No evidence of FQ biodegradation was observed but the pharmaceutical compounds adsorbed to the aerobic granular sludge, being gradually released into the medium in successive cycles after stopping the FQ feeding.Overall COD removal was not affected during the shock loadings. Activity of ammonia oxidizing bacteria and nitrite oxidizing bacteria did not seem to be inhibited by the presence of FQs (maximum of 0.03 and 0.01 mM for ammonium and nitrite in the effluent, respectively). However, during the FQs feeding, nitrate accumulation up to 1.7 mM was observed at the effluent suggesting that denitrification was inhibited. The activity of phosphate accumulating organisms was affected, as indicated by the decrease of P removal capacity during the aerobic phase. Exposure to the FQs also promoted disintegration of the granules leading to an increase of the effluent solid content, nevertheless the solid content at the bioreactor effluent returned to normal levels within ca. 1 month after removing the FQs in the feed allowing recovery of the bedvolume. Denaturing gradient gel electrophoresis revealed a dynamic bacterial community with gradual changes due to FQs exposure. Bacterial isolates retrieved from the granules predominantly belonged to α- and γ-branch of the Proteobacteria phylum.The capacity of the system to return to its initial conditions after withdrawal of the FQ compounds in the inlet stream, reinforced its robustness to deal with wastewaters containing organic pollutants.