Concentrations Of Azithromycin Research Articles

A New Electrochemical Sensor for the Detection of Azithromycin using Screen‐Printed Carbon Electrode Modified with Boron‐Doped Diamond Nanoparticles and Reduced‐Graphene Oxide

AbstractExcessive use of azithromycin (AZM) is associated with resistance and negative impacts on the environment. Therefore, this study aimed to develop an electrochemical sensor for AZM. The screen‐printed carbon electrode (SPCE) modified with boron‐doped diamond nanoparticle (BDDNP) and reduced‐graphene oxide (rGO) was successfully applied to measure the AZM concentration. The morphology of the modified electrode was characterized by scanning electron microscopy‐energy dispersive X‐ray (SEM‐EDX). Furthermore, the effect of electrochemical parameters, such as signal per background, scan rate, and selectivity was determined. The results showed that under optimum conditions, the plot of AZM demonstrated linearity in the range of 30–100 μM with a limit of detection (LOD) at 1.6 μM. The repeatability and recovery were also investigated, and the sensor was applied to real samples obtained from hospital wastewater. A remarkable 93.27 % recovery rate and excellent precision were recorded with an %RSD of 2.41 %. This study underscores the potential of SPCE modified with rGO/BDDNP for accurate and reliable measurement of AZM levels, showcasing great applicability in addressing the challenges associated with monitoring AZM in various environmental settings.

Pharmacokinetics and Bioequivalence of Two Formulations of Azithromycin Tablets: A Randomized, Single-Dose, Three-Period, Crossover Study in Healthy Chinese Volunteers Under Fasting and Fed Conditions.

Azithromycin is the first azalide antibiotic that is related to the macrolide family of antibiotics. Bioequivalence studies in China are initiated by the National Medical Products Administration (NMPA), which supports a generic consistency evaluation program for ensuring that generic products manufactured in China meet the required standards and provide equivalent therapeutic effects to their reference products. This study aimed to assess the bioequivalence of two azithromycin tablets under both fasting and fed conditions in healthy Chinese volunteers. This was a single-center, open-label, single-dose, randomized, three-way crossover trial with two independent groups (fasting group and fed group). A total of 72 healthy Chinese subjects (36 subjects in the fasting state and 36 subjects in the fed state) were enrolled and randomized to treatment. Blood samples were collected from 0 to 120 h after a single oral dose of a 250-mg generic azithromycin tablet (test, T) or branded azithromycin tablet (reference, R). The plasma concentrations of azithromycin were determined by high-performance liquid chromatography-tandem mass spectrometry (HPLC‒MS/MS). A non-compartmental analysis method was used to estimate the pharmacokinetic parameters. Adverse events were documented. In a fasting state, the bioequivalence of maximum plasma concentration (Cmax) was evaluated using the reference-scaled average bioequivalence (RSABE) approach (within-subject standard deviation, SWR >0.294), and the bioequivalence of area under the concentration-time curve from time 0 to the time of the last measurable plasma concentration (AUC0-t) and area under the concentration-time curve from time 0 extrapolated to infinity (AUC0-∞) were evaluated by the average bioequivalence (ABE) method (SWR < 0.294). The geometric mean ratio (GMR) of T/R for Cmax was 106.49%, while the 95% upper confidence bound was < 0. The GMRs of AUC0-t and AUC0-∞ were 103.34% and 101.28%, and the 90% confidence intervals (CIs) of the test/reference were 95.90-111.35%/94.85-108.15%, respectively. In the fed state, the RSABE approach was applied to estimate the bioequivalence of Cmax (SWR >0.294), and the ABE approach was applied to estimate the bioequivalence of AUC0-t and AUC0-∞ (SWR < 0.294). The GMR for Cmax was 99.80%, while the 95% upper confidence bound value was < 0. The GMRs of AUC0-t and AUC0-∞ were 97.07% and 98.15%, and the 90% CIs of the T/R were 90.02-104.68% and 90.66-106.25%, respectively. All adverse events were mild and transient. The trial indicated that the test and the reference azithromycin tablets were bioequivalent and well tolerated in healthy Chinese volunteers under both fasting and fed conditions. Clinicaltrials, ChiCTR2300071630 (retrospectively registered in 19/05/2023).

Minimum inhibitory concentrations of azithromycin in clinical isolates of Mycobacterium avium complex in Japan.

The latest guidelines include azithromycin as a preferred regimen for treating Mycobacterium avium complex (MAC) pulmonary disease. However, serially collected susceptibility data on clinical MAC isolates are limited, and no breakpoints have been determined. We investigated the minimum inhibitory concentrations (MICs) of azithromycin and clarithromycin for all MAC strains isolated in 2021 from a single center in Japan, excluding duplicates. The MICs were determined using a panel based on the microbroth dilution method, according to the latest Clinical and Laboratory Standards Institute recommendations. The MICs were determined for 318 MAC strains. Although there was a significant positive correlation between the MICs of azithromycin and clarithromycin, the MICs of azithromycin tended to be higher than those of clarithromycin. Among the cases in which the strains were isolated, 18 patients initiated treatment, including azithromycin treatment, after sample collection. Some patients infected with stains with relatively high azithromycin MICs achieved a microbiological cure with azithromycin-containing regimens. This study revealed a higher MIC distribution for azithromycin than clarithromycin, raising questions about the current practice of estimating azithromycin susceptibility based on the clarithromycin susceptibility test result. However, this was a single-center study that included only a limited number of cases treated with azithromycin. Therefore, further multicenter studies that include a greater number of cases treated with azithromycin are warranted to verify the distribution of azithromycin MICs and examine the correlation between azithromycin MICs and treatment effectiveness.IMPORTANCEThe macrolides serve as key drugs in the treatment of pulmonary Mycobacterium avium complex infection, and the administration of macrolide should be guided by susceptibility test results. Azithromycin is recommended as a preferred choice among macrolides, surpassing clarithromycin; however, drug susceptibility testing is often not conducted, and clarithromycin susceptibility is used as a surrogate. This study represents the first investigation into the minimum inhibitory concentration of azithromycin on a scale of several hundred clinical isolates, revealing an overall tendency for higher minimum inhibitory concentrations compared with clarithromycin. The results raise questions about the appropriateness of using clarithromycin susceptibility test outcomes for determining the administration of azithromycin. This study highlights the need for future discussions on the clinical breakpoints of azithromycin, based on large-scale clinical research correlating azithromycin susceptibility with treatment outcomes.

Population Pharmacokinetics and Individualized Medication of Azithromycin for Injection in Children Under 6 Years Old

Pharmacokinetic data for injectable azithromycin in children remain limited. This study aims to develop and validate a population pharmacokinetic model of azithromycin for injection in children under 6 years old and optimize its dosage regimen in this population. We prospectively enrolled patients under 6 years old who received azithromycin for injection at Beijing Friendship Hospital, Capital Medical University. Demographic information, clinical characteristics, and venous blood samples were collected in accordance with the research protocol. Azithromycin concentrations were determined using a validated UPLC-MS/MS method. The population pharmacokinetic model was structured using Phoenix NLME. The adequacy and robustness of the model was evaluated using VPC and bootstrap. We optimized azithromycin's dosing regimen for injection through Monte Carlo simulations. We included 254 plasma concentration data from 148 patients to establish the model. The clearance and volume were 1.27 L/h/kg and 45.6 L/kg, respectively. The covariates included were weight and age. VPC plots and nonparametric bootstrap showed that the final PPK model was reliable and robust. Based on Monte Carlo simulation, we derived a simple and practical dosing scheme. The results provided reference for individualized dosing in this population. The individualized dosing scheme based on Monte Carlo simulation can optimize clinical decision-making and guide personalized therapy.

Establishment and Evaluation of a Resazurin-Based Microdilution Assay for Microbial Sensitivity Test of Neisseria gonorrhoeae

To establish and evaluate a microbial sensitivity test method for Neisseria gonorrhoeae based on resazurin coloration. Based on the broth microdilution method, resazurin was added as a live bacteria indicator. WHO G, a WHO gonococcal reference strain, was used to optimize the incubation time for resazurin-stained bacteria and the color change was visually observed to obtain the results. Agar dilution method (the gold standard) and resazurin-based microdilution assay were used to determine the minimum inhibitory concentration (MIC) of azithromycin, ceftriaxone, and spectinomycin for 3 reference strains and 32 isolates of Neisseria gonorrhoeae. The results were analyzed based on essential agreement (EA), which reflected the consistency of the MIC values, category agreement (CA), which reflected the consistency in the determination of drug resistance, intermediary, and sensitivity, very major error (VME), which reflected false sensitivity, and major error (ME), which reflected pseudo drug resistance, to evaluate the accuracy of resazurin-based microdilution assay as a microbial sensitivity test of of Neisseria gonorrhoeae. CA and EA rates≥90% and VME and ME rates≤3% were found to be the acceptable performance rates. The results obtained 6 hours after resazurin was added were consistent with those of the agar dilution method and the resazurin-based microdilution assay was established accordingly based on this parameter. The EA of resazurin-based microdilution assay for measuring the MIC results of azithromycin, ceftriaxone, and spectinomycin was 97.1%, 91.5%, and 94.3%, respectively, and the CA was 88.6%, 94.3%, and 94.3%, respectively. The VME was 0% for all three antibiotics, while the ME was 11.4%, 5.7%, and 5.7%, respectively. The resazurin-based microdilution assay established in this study showed good agreement with agar dilution method for measuring the MIC of antibiotics against Neisseria gonorrhoeae. Moreover, the sensitivity results of this method were highly reliable and could be easily obtained through naked eye observation. Nonetheless, the results of drug resistance should be treated with caution and the optimization of parameters should be continued.

Brevicidine acts as an effective sensitizer of outer membrane-impermeable conventional antibiotics for Acinetobacter baumannii treatment.

The antibiotic resistance of Acinetobacter baumannii poses a significant threat to global public health, especially those strains that are resistant to carbapenems. Therefore, novel strategies are desperately needed for the treatment of infections caused by antibiotic-resistant A. baumannii. In this study, we report that brevicidine, a bacterial non-ribosomally produced cyclic lipopeptide, shows synergistic effects with multiple outer membrane-impermeable conventional antibiotics against A. baumannii. In particular, brevicidine, at a concentration of 1 μM, lowered the minimum inhibitory concentration of erythromycin, azithromycin, and rifampicin against A. baumannii strains by 32-128-fold. Furthermore, mechanistic studies were performed by employing erythromycin as an example of an outer membrane-impermeable conventional antibiotic, which showed the best synergistic effects with brevicidine against the tested A. baumannii strains in the present study. The results demonstrate that brevicidine disrupted the outer membrane of A. baumannii at a concentration range of 0.125-4 μM in a dose-dependent manner. This capacity of brevicidine could help the tested outer membrane-impermeable antibiotics enter A. baumannii cells and thereafter exert their antimicrobial activity. In addition, the results show that brevicidine-erythromycin combination exerted strong A. baumannii killing capacity by the enhanced inhibition of adenosine triphosphate biosynthesis and accumulation of reactive oxygen species, which are the main mechanisms causing the death of bacteria. Interestingly, brevicidine and erythromycin combination showed good therapeutic effects on A. baumannii-induced mouse peritonitis-sepsis models. These findings demonstrate that brevicidine is a promising sensitizer candidate of outer membrane-impermeable conventional antibiotics for treating A. baumannii infections in the post-antibiotic age.

Antimicrobial susceptibility of Treponema pallidum subspecies pallidum: an in-vitro study

The increasing incidence of syphilis and the limitations of first-line treatment with penicillin, particularly in neurosyphilis, neonatal syphilis, and pregnancy, highlight the need to expand the therapeutic repertoire for effective management of this disease. We assessed the in-vitro efficacy of 18 antibiotics from several classes on Treponema pallidum subspecies pallidum (T pallidum), the syphilis bacteria. Using the in-vitro culture system for T pallidum, we exposed the pathogen to a concentration range of each tested antibiotic. After a 7-day incubation, the treponemal burden was evaluated by quantitative PCR targeting the T pallidum tp0574 gene. The primary outcome was the minimum inhibitory concentration (MIC) at which the quantitative PCR values were not significantly higher than the inoculum wells. We also investigated the susceptibility of macrolide-resistant strains to high concentrations of azithromycin, and the possibility of developing resistance to linezolid, a proposed candidate for syphilis treatment. Amoxicillin, ceftriaxone, several oral cephalosporins, tedizolid, and dalbavancin exhibited anti-treponemal activity at concentrations achievable in human plasma following regular dosing regimens. The experiments revealed a MIC for amoxicillin at 0·02 mg/L, ceftriaxone at 0·0025 mg/L, cephalexin at 0·25 mg/L, cefetamet and cefixime at 0·0313 mg/L, cefuroxime at 0·0156 mg/L, tedizolid at 0·0625 mg/L, spectinomycin at 0·1 mg/L, and dalbavancin at 0·125 mg/L. The MIC for zoliflodacin and balofloxacin was 2 mg/L. Ertapenem, isoniazid, pyrazinamide, and metronidazole had either a poor or no effect. Azithromycin concentrations up to 2 mg/L (64 times the MIC) were ineffective against strains carrying mutations associated to macrolide resistance. Exposure to subtherapeutic doses of linezolid for 10 weeks did not induce phenotypic or genotypic resistance. Cephalosporins and oxazolidinones are potential candidates for expanding the current therapeutic repertoire for syphilis. Our findings warrant testing efficacy in animal models and, if successful, clinical assessment of efficacy. European Research Council.

Development of a national antibiotic multimetric index for identifying watersheds vulnerable to antibiotic pollution

Improved surveillance of antibiotics and antibiotic resistance (AR) throughout the environment is an important aspect of the prevention and control of threats posed to human and ecological health. In response to field investigations often limited by resources and time, this study aims to develop a systematic approach to assess watershed vulnerability to antibiotic pollution and AR by integrating modeling and field studies. The national antibiotic pollution vulnerability index was developed to identify watersheds most impacted by antibiotic sources. The index incorporates multiple metrics representing antibiotic pollution driven by both agricultural activities and municipal wastewater (i.e. outpatient antibiotic prescriptions, wastewater treatment plant effluent flow, stream order and dilution factor of effluent-receiving streams, manure application, and animal facilities), alongside climate change indicators (i.e., temperature, precipitation, and runoff). The pollution index was applied at a state level in North Carolina to identify the most-impacted watersheds and inform site selection for targeted field study quantifying azithromycin, ciprofloxacin, sulfamethoxazole, and trimethoprim concentrations. Modeled-informed sites in NC demonstrated the highest reported concentrations of azithromycin, trimethoprim, and sulfamethoxazole compared to previous NC studies, confirming the index effectiveness in identifying watersheds with higher antibiotic concentrations. At the national scale, watersheds relatively more vulnerable to antibiotic pollution are predominantly located in the Midwest, South, and Northeast regions of the U.S., with Iowa and Indiana being the most impacted states. Climate change is expected to exacerbate watershed vulnerability to agriculture-driven AR in the Midwest and Northeast due to an increase in precipitation and mean temperature coupled with intense agricultural activities. In addition, due to climate change-induced reductions in precipitation and runoff, watersheds in the Midwest, Mid-Atlantic, and South Central are dominantly at higher risk of effluent-driven AR occurrences. We have disseminated the developed indices as open-source online tools to aid in prioritizing strategies to mitigate AR occurrence across the U.S.

Impact of azithromycin, doxycycline and redox-active small molecules on amoxicillin-induced Chlamydia pneumoniae persistence

Amoxicillin is recommended as primary treatment for community-acquired bacterial pneumonia (CABP). 5–10% of CABP cases are caused by Chlamydia pneumoniae, an obligate intracellular bacterium which responds to beta-lactam antibiotics by converting to a persistent phenotype. To support rational pharmacotherapy of C. pneumoniae infections, we investigated how clinically relevant concentrations of azithromycin and doxycycline affect amoxicillin induced C. pneumoniae persistence. Given the known role of redox state alterations in the action of bactericidal antibiotics and widespread use of redox-active dietary supplements when experiencing respiratory symptoms, we also studied how redox active compounds affect the studied antibiotic treatments. Our data demonstrate that clinically applied amoxicillin concentrations (10 and 25 mg/l) fail to eradicate C. pneumoniae infection in respiratory epithelial cells. Transmission electron microscopy (TEM) of amoxicillin-treated C. pneumoniae infected cells reveal aberrant bacterial morphology characteristic of chlamydial stress response. Amoxicillin was also found to significantly limit the antichlamydial effect of azithromycin or doxycycline. However, based on quantitative culture and quantitative PCR data, azithromycin was superior to doxycycline in C. pneumoniae eradication either as monotherapy or in combination with amoxicillin. Amoxicillin was also found to decrease respiratory epithelial cell glutathione (GSH) levels, whereas redox-active dibenzocyclooctadiene lignans increased C. pneumoniae load in amoxicillin-treated cultures up to two-fold. These data highlight the impact of relative administration time on the efficacy of antichlamydial antibiotics and indicate unfavorable interactions between amoxicillin and redox-active small molecules.

Azithromycin induces neurotoxicity in zebrafish by interfering with the VEGF/Notch signaling pathway

Azithromycin (AZM) is a widely used antibiotic in both human and veterinary medicine, and its use has significantly increased during the COVID-19 pandemic. However, potential adverse effects of AZM on aquatic organisms have not been well studied. In this study, we explored the neurotoxicity of AZM in zebrafish and delved into its underlying mechanisms. Our results showed that AZM exposure resulted in a spectrum of detrimental effects in zebrafish, encompassing abnormal behaviors, damaged neuronal development, aberrant lateral line nervous system development, vascular malformations and perturbed expression of genes related to neural development. Moreover, we observed a concentration-dependent exacerbation of these neurotoxic manifestations with increasing AZM concentrations. Notably, AZM induced excessive cell apoptosis and oxidative stress damage. In addition, alterations in the expression levels of the genes involved in the VEGF/Notch signaling pathway were evident in AZM-exposed zebrafish. Consequently, we hypothesize that AZM may induce neurotoxicity by influencing the VEGF/Notch signaling pathway. To validate this hypothesis, we introduced a VEGF signaling inhibitor, axitinib, and a Notch signaling agonist, valproic acid, alongside AZM exposure. Remarkably, the administration of these rescue compounds significantly mitigated the neurotoxic effects induced by AZM. This dual verification provides compelling evidence that AZM indeed induces neurotoxicity during the early developmental stages of zebrafish, primarily through its interference with the VEGF/Notch pathway. Innovatively, our study reveals the molecular mechanism of AZM-induced neurotoxicity from the perspective of the close connection between blood vessels and nervous system. These findings provide new insights into the potential mechanisms underlying the neurotoxic effect of antibiotics and highlight the need for further investigation into the ecotoxicological effects of antibiotics on aquatic organisms and the potential risks to human health.

Antibacterial activity of the novel compound Sudapyridine (WX-081) against Mycobacterium abscessus.

This study aimed to investigate sudapyridine (WX-081) antibacterial activity against Mycobacterium abscessus in vitro and its effect on in vivo bacterial growth and host survival using a zebrafish model of M. abscessus infection. WX-081 in vitro antibacterial activity was assessed based on growth inhibition of M. abscessus standard strain ATCC19977 and 36 clinical isolates. Maximum tolerated concentrations (MTCs) of WX-081, bedaquiline, and azithromycin and inhibition of M. abscessus growth were assessed in vivo after fluorescently labelled bacilli and drugs were injected into zebrafish. Bacterial counts were analysed using one-way ANOVA and fluorescence intensities of zebrafish tissues were analysed and expressed as the mean ± SE. Moreover, Kaplan-Meier survival analysis was conducted to assess intergroup differences in survival of M. abscessus-infected zebrafish treated with different drug concentrations using a log-rank test, with a p value <0.05 indicating a difference was statistically significant. Drug sensitivity testing of M. abscessus standard strain ATCC19977 and 36 clinical isolates revealed MICs ranging from 0.12-0.96 µg/mL and MIC50 and MIC90 values of 0.48 µg/mL and 0.96 µg/mL, respectively. Fluorescence intensities of M. abscessus-infected zebrafish tissues was lower after treatment with the WX-081 MTC (62.5 µg/mL) than after treatment with the azithromycin MTC (62.5 µg/mL) and the bedaquiline MTC (15.6 µg/mL). When the concentration of WX-081 increased from 1.95µg/mL to 1/8 MTC(7.81µg/mL), the survival rate of zebrafish at 4-9 dpf decreased from 90.00% to 81.67%. WX-081 effectively inhibited M. abscessus growth in vitro and in vivo and prolonged survival of M. abscessus-infected zebrafish, thus indicating that WX-081 holds promise as a clinical treatment for M. abscessus infection.

Wastewater surveillance of pharmaceuticals during the COVID-19 pandemic in Mexico City and the Mezquital Valley: A comprehensive environmental risk assessment

This study tracked five pharmaceutically active compounds (PhACs) in Mexico City's sewage, namely, famotidine, indomethacin, dexamethasone, azithromycin, and ivermectin, which were used to treat COVID-19. The monitoring campaign was carried out over 30 months (May 2020 to November 2022), covering the five COVID-19 waves in Mexico. In the Central Emitter, the main sewage outflow, famotidine displayed levels of 132.57 ± 28.16 ng L−1 (range from < LOQ to 189.1 ng L−1), followed by indomethacin (average 672.46 ± 116.4 ng L−1, range from 516.7 to 945.2 ng L−1), dexamethasone (average 610.4 ± 225.7 ng L−1, range from 233.4 to 1044.5 ng L−1), azithromycin (average 4436.2 ± 903.6 ng L−1, range from 2873.7 to 5819.6 ng L−1), and ivermectin (average 3413.3 ± 1244.6 ng L−1, range from 1219.8 to 4622.4 ng L−1). The concentrations of dexamethasone, azithromycin and ivermectin were higher in sewage from a temporary COVID-19 care unit, by a factor of 3.48, 3.52 and 2.55, respectively, compared with those found in municipal wastewater. In the effluent of the Atotonilco Wastewater Treatment Plant (AWWTP), which treats near 60 % of the Mexico City's sewage, famotidine was absent, while concentrations of indomethacin, dexamethasone, azithromycin and ivermectin were 78.2 %, 76.7 %, 74.4 %, and 88.1 % lower than those in the influent, respectively. The occurrence of PhACs in treated and untreated wastewater resulted in medium to high environmental risk since Mexico City's wastewater is reused for irrigation in the Mezquital Valley. There, PhACs were found in irrigation canals at lower levels than those observed in Mexico City throughout the monitoring. On the other hand, famotidine, indomethacin, and dexamethasone were not found in surface water resulting from the infiltration of wastewater through soil in Mezquital Valley, while azithromycin and ivermectin sporadically appeared in surface water samples collected through 2021. Using an optimized risk assessment based on a semi-probabilistic approach, the PhACs were prioritized as ivermectin > azithromycin > dexamethasone > famotidine > indomethacin.

Evaluation of the Ecological Risk and the Development of Antimicrobial Resistance due to the Presence of the Macrolide Antibiotics Azithromycin and Clarithromycin in Romanian Aquatic Environment

The presence of antibiotics in the aquatic environment can result in the emergence of antibiotic-resistant pathogens. In this paper, our aim was to identify, quantify and distribute macrolide antibiotics in the aquatic environment in the river basins of Arges-Vedea, Buzau-Ialomita and Dobrogea-Litoral and of the Danube River. In the Arges-Vedea river basin area, clarithromycin was detected most frequently, i.e. 58.62%, in the Buzau-Ialomita river basin area with a frequency of 92.31% azithromycin was detected, and in the Dobrogea - Litoral river basin area clarithromycin was detected with a frequency of 35.48%. The highest concentration of azithromycin, 559 ng/L and of clarithromycin, 502 ng/L was recorded in the Buzau-Ialomita river basin. The ecotoxicological RQecotox risk was also estimated, as well as the risk of developing antibiotic resistance of RQAMR pathogens, by the ratio of Measured Environmental Concentration (MEC) and Predicted No Effect Concentrations (PNEC). The study also aimed to investigate the prevalence of antibiotic resistance in potentially pathogenic bacteria isolated from aquatic environments. Thus, for β-lactam antibiotics, E. coli strains isolated from the Danube River exhibited a very high level of resistance to ampicillin (51%) and high level to azithromycin (43%), cefazolin (38%), amoxicillin+clavulanic acid (36%) and cefoxitin (26%). Low and respectively, low level resistance was noticed for aztreonam (6%) and imipenem (4%).

Poly(sebacic acid) microparticles loaded with azithromycin as potential pulmonary drug delivery system: Physicochemical properties, antibacterial behavior, and cytocompatibility studies

Recurrent bacterial infections are a common cause of death for patients with cystic fibrosis and chronic obstructive pulmonary disease. Herein, we present the development of the degradable poly(sebacic acid) (PSA) microparticles loaded with different concentrations of azithromycin (AZ) as a potential powder formulation to deliver AZ locally to the lungs. We characterized microparticle size, morphology, zeta potential, encapsulation efficiency, interaction PSA with AZ and degradation profile in phosphate buffered saline (PBS). The antibacterial properties were evaluated using the Kirby-Bauer method against Staphylococcus aureus. Potential cytotoxicity was evaluated in BEAS-2B and A549 lung epithelial cells by the resazurin reduction assay and live/dead staining. The results show that microparticles are spherical and their size, being in the range of 1–5 μm, should be optimal for pulmonary delivery. The AZ encapsulation efficiency is nearly 100 % for all types of microparticles. The microparticles degradation rate is relatively fast – after 24 h their mass decreased by around 50 %. The antibacterial test showed that released AZ was able to successfully inhibit bacteria growth. The cytotoxicity test showed that the safe concentration of both unloaded and AZ-loaded microparticles was equal to 50 μg/ml. Thus, appropriate physicochemical properties, controlled degradation and drug release, cytocompatibility, and antibacterial behavior showed that our microparticles may be promising for the local treatment of lung infections.

High-risk antibiotics positively correlated with antibiotic resistance genes in five typical urban wastewater

Antibiotic resistance genes (ARGs) and antibiotic amount increased within close proximity to human dominated ecosystems. However, few studies assessed the distribution of antibiotics and ARGs in multiple ecosystems especially the different urban wastewater. In this study, the spatial distribution of ARGs and antibiotics across the urban wastewater included domestic, livestock, hospital, pharmaceutical wastewater, influent of the wastewater treatment plant (WWTP) in Northeast China. The q-PCR results showed that ARGs were most abundant in community wastewater and followed by WWTP influent, livestock wastewater, pharmaceutical wastewater and hospital wastewater. The ARG composition differed among the five ecotypes with qnrS was the dominant ARG subtypes in WWTP influent and community wastewater, while sul2 dominant in livestock, hospital, pharmaceutical wastewater. The concentration of antibiotics was closely related to the antibiotic usage and consumption data. In addition to the high concentration of azithromycin at all sampling points, more than half of the antibiotics in livestock wastewater were veterinary antibiotics. However, antibiotics that closely related to humankind such as roxithromycin and sulfamethoxazole accounted for a higher proportion in hospital wastewater (13.6%) and domestic sewage (33.6%), respectively. The ambiguous correlation between ARGs and their corresponding antibiotics was detected. However, antibiotics that exhibited high ecotoxic effects were closely and positively correlated with ARGs and the class 1 integrons (intI1), which indicated that high ecotoxic compounds might affect antimicrobial resistance of bacteria by mediating horizontal gene transfer of ARGs. The coupling mechanism between the ecological risk of antibiotics and bacterial resistance needed to be further studied, and thereby provided a new insight to study the impact of environmental pollutants on ARGs in various ecotypes.

Reporting of Azithromycin Activity against Clinical Isolates of Extensively Drug-Resistant Salmonella enterica Serovar Typhi.

This study aimed to evaluate the minimum inhibitory concentration (MIC) of azithromycin (AZM) in clinical isolates of extensively drug-resistant (XDR) Salmonella Typhi (i.e., resistant to chloramphenicol, ampicillin, trimethoprim-sulfamethoxazole, fluoroquinolones, and third-generation cephalosporin) using the E-test versus the broth microdilution method (BMD). From January to June 2021, a retrospective cross-sectional study was carried out in Lahore, Pakistan. Antimicrobial susceptibility was performed initially by the Kirby-Bauer disk diffusion method for 150 XDR Salmonella enterica serovar Typhi isolates, and MICs of all the recommended antibiotics were determined by the VITEK 2 (BioMérieux) fully automated system using Clinical Laboratory Standard Institute (CLSI) 2021 guidelines. The E-test method was used to determine AZM MICs. These MICs were compared with the BMD, which is the method recommended by the CLSI but not adopted in routine laboratory reporting. Of 150 isolates, 10 (6.6%) were resistant by disk diffusion. Eight (5.3%) of these had high MICs against AZM by the E-test. Only three isolates (2%) were resistant by E-test, having an MIC of 32 μg/mL. All eight isolates had a high MIC by BMD with different MIC distributions, but only one was resistant, having an MIC of 32 μg/mL by BMD. The sensitivity, specificity, negative predictive value, positive predictive value, and diagnostic accuracy of the E-test method versus BMD were 98.65%,100%, 99.3%, 33.3%, and 98.6%, respectively. Similarly, the concordance rate was 98.6%, negative percent agreement was 100%, and positive percent agreement was 33%. The BMD is the most reliable approach for reporting AZM sensitivity in XDR S. Typhi compared with the E-test and disk diffusion methods. Potentially, AZM resistance in XDR S. Typhi is around the corner. Sensitivity patterns should be reported with MIC values, and if possible, higher values should be screened for the presence of any potential resistance genes. Antibiotic stewardship should be strictly implemented.

Effect of Azithromycin on Genotoxicity in Allium Test

The effect of azithromycin at a concentration of 50-1000.0 mg/l on the pathology of mitosis in the Allium test in the first metaphase after treatment and after the completion of the recovery period is being investigated. Research methods: Allium test, cytogenetic analysis, statistical analysis. The Allium test was conducted on a batch of the Stuttgarter variety which is genetically unstable as the result of micronucleus test. The results of the analysis showed that the tested concentrations of azithromycin (50.0-1000.0 mg/l) in the first mitosis did not cause a significant increase in the values of mitosis pathology (PM) compared with the negative control. However, there was a significant increase (8 times) in the number of cells with PM in the “aftereffect, after 48 hours” variant for a concentration of 300.0 mg/l of azithromycin: from 4.1% to 32.3%. It should be noted that under the action of all tested concentrations of azithromycin, the same tendency is observed as in the control variant in terms of the preservation of the PM spectrum and the dominance, firstly, of the pathology “overrun/lag of chromosomes”, and secondly, the pathology of “asynchronous spindle fission”. The percentage of mentioned PMs ranges from 52 to 74, and from 7 to 22, respectively. Other types of PMs, according to their occurrence in experimental variants, are arranged in the following sequence: c-mitosis, chromosome scattering, chromosome outliers, polyploidy, chromosome stickiness, chromosomal and chromatid bridges, pycnosis of interphase nuclei. Under the action of azithromycin, starting from a concentration of 100.0 mg/l to 1000.0 mg/l, polyploid cells were observed in dividing cells, which contained a tetraploid set of chromosomes. In experimental variants in single cells, a significant increase in the size of the nuclei of interphase cells and the presence of nuclear buds were noted, which is considered as markers of polyploidization and gene amplification. The recovery period (after 48 hours after the action of the antibiotic) for azithromycin does not reduce the number of cells with PM but changes their composition and spectrum.

How toxic is the COVID-19 drug azithromycin in the presence of Posidonia oceanica? Toxicokinetics and experimental approach of meiobenthic nematodes from a metallically pristine area

The current study presents the results of an experiment carried to assess the impact of azithromycin, a COVID-19 drug, probably accumulated in marine sediments for three years, since the start of the pandemic, on benthic marine nematodes. It was explored the extent to which a common macrophyte from the Mediterranean Sea influenced the toxic impact of azithromycin on meiobenthic nematodes. Metals are known to influence toxicity of azithromycin. The nematofauna from a metallically pristine site situated in Bizerte bay, Tunisia, was exposed to two concentrations of azithromycin [i.e. 5 and 10 μg l−1]. In addition, two masses of the common macrophyte Posidonia oceanica [10 and 20% Dry Weight (DW)] were considered and associated with azithromycin into four possible combinations. The abundance and the taxonomic diversity of the nematode communities decreased significantly following the exposure to azithromycin, which was confirmed by the toxicokinetic data and behaving as substrate for P-glycoprotein (P-gp). The toxicity of 5 μg l−1 dosage of azithromycin was partially reduced at 10% DW of Posidonia and completely at 20% DW. The results showed that 5 μg l−1 of azithromycin can be reduced by the macrophyte P. oceanica when present in the environment at low masses as 10% DW.

Biotesting of Chlorella sp algae for certain medicinal drugs

In recent years, the study of drugs as pollutants in the country is gaining momentum. This is because the country does not use special wastewater treatment devices for pharmaceuticals. They enter the environment in different ways. Therefore, research on biotesting drugs in living organisms is underway. The article presents the results of experiments conducted to assess the effect of paracetamol, azithromycin, suprastin on the number and biomass of Chlorella sp algae. Chlorella sp algae are a widely used organism for biotesting. Among the above drugs in the study, paracetamol had a negative effect on the number and biomass of algae. The minimum harmful concentration of paracetamol for chlorella was 5 mg/L, the minimum harmful concentration of azithromycin for chlorella was 200 mg/L, the minimum harmful concentration of chlorella suprastin was 8 mg/L. When comparing the effect of paracetamol, azithromycin, suprastin on the number of green algae Chlorella sp, it was observed that paracetamol had the highest effect. Concentration of paracetamol 10 mg/L brought the number of chlorella close to 0. High concentrations of paracetamol and suprastinin the biomass of chlorella showed contamination of the environment.

ANALYSIS OF MINIMAL INHIBITORY CONCENTRATION(MIC) OF AZITHROMYCIN IN EXTREMELY DRUG RESISTANT (XDR)SALMONELLA ENTERICA SEROVAR TYPHI IN PRE-COVID AND AFTER ONE YEAR OF COVID-19 PANDEMIC

Abstract &#x0D; Objective: To compare the Minimal Inhibitory Concentration (MIC) of Azithromycin in Extremely Drug Resistant (XDR) Salmonella enterica serovar Typhi (S. Typhi), Pre-COVID and after one year of COVID-19 pandemic.&#x0D; Methodology: This cross-sectional study was conducted in the Department of Microbiology, Combined Military Hospital, Lahore, Pakistan, from May 2021 to April 2022. All XDR S. Typhi isolated during the study period were included and tested. Kirby –Bauer Disk Diffusion method was used for antibiotic susceptibility, and Azithromycin MIC was determined by the E-strip method. The latest Clinical &amp; Laboratory Standards Institute (CLSI) breakpoints for Azithromycin susceptibility testing were used. &#x0D; Results: A total of 384 XDR S. Typhi were included in the study. MICs of these isolates were in the ranges of 2-16ug/mL, and all were sensitive to Azithromycin as per CLSI standards. These were further compared with MICs of the pre-covid isolates. The MICs of the isolates were relatively higher as compared to pre-COVID MICs. &#x0D; Conclusion: The potential Azithromycin resistance in XDR Salmonella Typhi is knocking at the door. Sensitivity patterns should be reported with MICs values, and if possible higher values should be screened for the presence of any potential resistance genes. Antibiotic stewardship should be strictly implemented.