Lyophilized Human Bone Allograft as an Antibiotic Carrier: An In Vitro and In Vivo Study.

Background: Antibiotics delivered from implanted bone substitute materials (BSM) can potentially be used to prevent acute infections and biofilm formation, providing high concentrations of antibiotics at the surgical site without systemic toxicity. In addition, BSM should allow osteoconductivity supporting bone healing without further surgery. Promising results have been achieved using lyophilized bone allografts mixed with antibiotics. Methods: In this study specially prepared human bone allografts were evaluated as an antibiotic carrier in vitro and in vivo. The efficacy of different antibiotic-impregnated bone allografts was measured by drug release tests in vitro and in vivo and bacterial susceptibility tests using four bacterial species usually responsible for implant-associated infections. Results: The loading procedures of allograft bone substitutes with antibiotics were successful. Some of the antibiotic concentrations exceeded the MIC90 for up to 7 days in vitro and for up to 72 h in vivo. The susceptibility tests showed that S. epidermidis ATCC 12228 was the most susceptible bacterial species in comparison to the other strains tested for all antibiotic substances. Vancomycin and rifampicin showed the best results against standard and patient-isolated strains in vitro. In vivo, new bone formation was comparable in all study groups including the control group without antibiotic loading. Conclusions: Human bone allografts showed the capacity to act as customized loaded antibiotic carriers to prevent acute infections and should be considered in the management of bone infections in combination with systemic antimicrobial therapy.

Establishment of Epidemiological Resistance Cut-Off Values of Aquatic Aeromonas to Eight Antimicrobial Agents.

The abuse of antibiotics in aquaculture has led to the increasing rate of antibiotic resistance of aquatic bacteria including Aeromonas, which is an increasing threat to environmental and human health. To date, no epidemiological cut-off values (COWT) for Aeromonas spp. have been established by the Clinical and Laboratory Standards Institute nor the European Commission on Antimicrobial Susceptibility Testing. In this study, commercially prepared minimum inhibitory concentration (MIC) test 96-well plates (dry-form plates) were used to determine the MIC of eight antimicrobial agents against 556 Aeromonas strains. The obtained MIC distributions were simulated and analyzed by NRI and ECOFFinder to obtain tentative COWT values for Aeromonas spp. The COWT values of eight kinds of representative antimicrobial agents including trimethoprim-sulfamethoxazole, erythromycin, doxycycline, neomycin, colistin, florfenicol, enrofloxacin, and ceftazidime for Aeromonas spp. were established and were 0.25, 64/32, 4/2, 8, 4, 1, 0.062/0.125, and 0.5 µg/mL, respectively. Results showed that Aeromonas spp. had a very high proportion of non-wild-type strains to enrofloxacin, florfenicol, and doxycycline, which are the most widely used antimicrobials in aquaculture. The COWT values for Aeromonas spp. obtained in this study can contribute to the final establishment of COWT for Aeromonas spp. internationally.

Clinical Distribution Characteristics of 1439 Carbapenem-Resistant Escherichia coli Strains in China: Drug Resistance, Geographical Distribution, Antibiotic MIC50/90.

PURPOSE: To explore the clinical distribution characteristics and antimicrobial susceptibilities of carbapenem-resistant Escherichia coli (CR-ECO) in Hebei Province, China, from 2017 to 2019, and provide data on the treatment of this bacterial infection and the prevention of its spread. MATERIALS AND METHODS: A total of 1439 CR-ECO strains were collected from 2017 to 2019 in Hebei Province, China. Drug sensitivity tests were performed using the minimum inhibitory concentration (MIC) method, and the data were analyzed statistically using WHONET5.6 software. RESULTS: A total of 54,377 strains of Escherichia coli were isolated in Hebei Province from 2017 to 2019, of which 1439 strains were CR-ECO (2.65%). The highest proportion (33.78%) of strains was isolated from urine, and the detection rate showed a slow downward trend over the past 3 years. CR-ECO was mainly detected in densely populated and economically developed areas. Of all the patients, 54.2% were from the medical ward; the ratio of male to female patients with CR-ECO infections was 1.35:1; elderly patients and adults accounted for 59.6% and 30.8%, respectively, whereas minors and newborns accounted for 4.9% and 4.7%, respectively. For CR-ECO, the drug resistance rates to ß-lactams were all higher than 80% and there was an annual increasing trend, while the drug resistance rates to quinolones remained nearly unchanged. The rate of resistance to aminoglycosides was relatively low, especially to amikacin (approximately 22%). The MIC50 of other antibacterial drugs, except amikacin, was equal to or higher than the break point of drug resistance. CONCLUSION: From 2017 to 2019, the isolation rate of CR-ECO in Hebei Province, China, remained stable; however, the drug resistance rate showed an upward trend, primarily in cases of urinary tract infections in older men; the resistance rate to amikacin was the lowest.

In Vitro Antimicrobial Activity of Quinolones Against Major Bacterial Isolates from the Ocular Surface Bacterial Flora of Tertiary Hospital Patients in Japan.

Purpose: To determine the in vitro antimicrobial activity of quinolones against major bacterial isolates from the ocular surface bacterial flora of patients in a tertiary hospital for selection of optimal antibiotic eye drop during the perioperative stage. Methods: The conjunctival sac scraping of 933 patients who underwent ophthalmic surgery was cultivated and bacterial species of the isolates were identified. The minimum inhibitory concentrations (MICs) of gatifloxacin (GFLX), moxifloxacin (MFLX), levofloxacin (LVFX), and tosufloxacin (TFLX) were measured by microdilution methods. The cumulative percentages of MICs of 4 quinolones against major bacteria were calculated. The concentrations of quinolones inhibiting 50% (MIC50) and 90% (MIC90) of the major bacteria were compared. Results: The study mainly included 784 patients scheduled for cataract surgery, 73 for vitrectomy, 30 for corneal transplantation, 30 for conjunctival surgery, 11 for eyelid surgery. The most frequently isolated bacterium was coagulase-negative Staphylococci (CNS) (184 strains), followed by Corynebacterium (107 strains), Staphylococcus aureus (33 strains), Streptococcus (18 strains), and Enterococcus (13 strains). The percentages of methicillin-sensitive CNS isolates for which MIC of GFLX, MFLX, LVFX, and TFLX was 0.06 µg/mL or less were 8.0%, 13.4%, 5.4%, and 63.4%, respectively. Similarly, the percentage for Corynebacterium was 23.0%, 23.0%, 0%, and 35.6%, respectively. MIC50 of TFLX for Streptococcus and Enterococcus showed the lowest values, 0.12 and 0.25 µg/mL, respectively. Conclusions: Among 4 quinolones, TFLX has the highest in vitro antimicrobial activity against major bacterial isolates from the ocular surface bacterial flora of patients in a tertiary hospital.

Variations in the C-unit of bedaquiline provides analogues with improved biology and pharmacology.

Analogues of the anti-tuberculosis drug bedaquiline, bearing a 3,5-dimethoxy-4-pyridyl C-unit, retain high anti-bacterial potency yet exert less inhibition of the hERG potassium channel, in vitro, than the parent compound. Two of these analogues (TBAJ-587 and TBAJ-876) are now in preclinical development. The present study further explores structure-activity relationships across a range of related 3,5-disubstituted-4-pyridyl C-unit bedaquiline analogues of greatly varying lipophilicity (clogP from 8.16 to 1.89). This broader class shows similar properties to the 3,5-dimethoxy-4-pyridyl series, being substantially more potent in vitro and equally active in an in vivo (mouse) model than bedaquiline, while retaining a lower cardiovascular risk profile through greatly attenuated hERG inhibition.

Antimicrobial susceptibility pattern of oral isolates of Aggregatibacter actinomycetemcomitans.

BACKGROUND: Aggregatibacter actinomycetemcomitans is involved in the etiology of localized aggressive periodontitis (LAP), a condition that frequently requires supplemental antibiotic therapy. Information on antimicrobial susceptibility pattern and guidelines for oral antibiotic therapy are not available on Indian patients. AIM: The main aim of the present study was to screen clinical isolates on a panel of antibiotics commonly used for oral/systemic therapy. MATERIALS AND METHODS: The study included 40 strains of A. actinomycetemcomitans isolated from patients with LAP. The subgingival plaque was plated onto Trypticase Soy Serum Bacitracin Vancomycin Agar medium and incubated for 72 h, and suspected colonies were confirmed by phenotypic tests. Each isolate was tested against a panel of 12 antibiotics using MIC gradient strip test. ATCC strains of A. actinomycetemcomitans serotype A and C were used as standards. Performance and interpretation of the test were done according to the manufacturers' instructions. Distribution of MICs among isolates (n = 40) were used to calculate concentrations inhibiting 50% (MIC50) and 90% (MIC90) of strains. RESULTS: Moxifloxacin, cefotaxime and ceftriaxone showed excellent activity with 100% growth inhibition followed by amoxicillin, amoxiclav and doxycycline (>90% activity). The bacterial strains were moderately susceptible to cefuroxime, cefazolin and tetracycline but displayed poor susceptibility to clindamycin and azithromycin. All isolates were resistant to metronidazole. CONCLUSION: The isolates of A. actinomycetemcomitans displayed a high level of resistance to azithromycin and clindamycin. Development of resistance against tetracycline also appears to be significant. Variable resistance among the different members of the cephalosporin group is a factor to be investigated further since susceptibility profile against these antibiotics and interpretative criteria for oral bacteria are not available.

3,5-Dialkoxypyridine analogues of bedaquiline are potent antituberculosis agents with minimal inhibition of the hERG channel.

Bedaquiline is a new drug of the diarylquinoline class that has proven to be clinically effective against drug-resistant tuberculosis, but has a cardiac liability (prolongation of the QT interval) due to its potent inhibition of the cardiac potassium channel protein hERG. Bedaquiline is highly lipophilic and has an extremely long terminal half-life, so has the potential for more-than-desired accumulation in tissues during the relatively long treatment durations required to cure TB. The present work is part of a program that seeks to identify a diarylquinoline that is as potent as bedaquiline against Mycobacterium tuberculosis, with lower lipophilicity, higher clearance, and lower risk for QT prolongation. Previous work led to the identification of compounds with greatly-reduced lipophilicity compounds that retain good anti-tubercular activity in vitro and in mouse models of TB, but has not addressed the hERG blockade. We now present compounds where the C-unit naphthalene is replaced by a 3,5-dialkoxy-4-pyridyl, demonstrate more potent in vitro and in vivo anti-tubercular activity, with greatly attenuated hERG blockade. Two examples of this series are in preclinical development.

Assessing concentration of antibiotics in tissue during oral treatments against piscirickettsiosis.

The use of antimicrobials in aquaculture is increasingly being scrutinized. In Chile, piscirickettsiosis accounts for approximately 90% of the total volume of antibiotics used in marine aquaculture. Treatment failures are frequently reported, but there is limited information on why this occurs. Fish producers have started assessing the level of antibiotics in fish tissues during and immediately after in-feed treatments to determine if they are adequately medicating their fish. In this study, we evaluated the probability of finding antibiotic concentrations in muscle tissue above the minimum inhibitory concentration for 90% of the P. salmonis isolates (MIC90) recently tested in Chile, for two antibiotics commonly used in aquaculture. We found that the proportion of fish with antibiotic concentrations above the MIC90 varied, depending on the product used, species, day of sample collection, and size category of fish within a cage. The proportion of fish above the MIC90 was lower in fish treated with florfenicol than in fish treated with oxytetracycline. Using a mixed-effects logistic model, we modeled the probability of antibiotic concentrations above MIC90 when fish were treated with florfenicol. Our model suggested lower probabilities of having concentrations above MIC90 in Atlantic salmon than in rainbow trout when samples were collected 14 days after the treatment started compared to 7 days, and in the smaller fish within a cage. We discuss these findings and hypothesize about potential issues with treating large populations of fish with in-feed antimicrobials.

Amoxicillin concentrations in relation to beta-lactamase activity in sputum during exacerbations of chronic obstructive pulmonary disease.

BACKGROUND: Acute exacerbations of chronic obstructive pulmonary disease (COPD) are often treated with antibiotics. Theoretically, to be maximally effective, the antibiotic concentration at sites of infection should exceed the minimum inhibitory concentration at which 90% of the growth of potential pathogens is inhibited (MIC90). A previous study showed that most hospitalized COPD patients had sputum amoxicillin concentrations

Comparison of aqueous humour concentration after single high dose versus multiple administration of topical moxifloxacin in rabbits.

For the prevention of postoperative ocular infections prophylactic topical antibiotics are routinely used. Studies evaluating comparative difference between single dose versus multiple dose administration on aqueous humour concentration of moxifloxacin are lacking. This study compared the aqueous humour concentration of moxifloxacin following its topical administration in rabbit eyes with two dose regimens. Twelve albino rabbits were divided into two groups. In group-1, two drops were administered thrice (total six drops) at 2 min intervals, in both the eyes; in group-2, two drops of moxifloxacin were administered three times a day for three days and also two h before aqueous humour collection i.e. on fourth day. Mean aqueous humour concentrations were calculated and compared using Student's 't' test and P<0.05 was considered significant. Moxifloxacin concentration in aqueous humour in group-1 was 23.79 µg/ml and in group-2 was 42.08 µg/ml. Both dosing regimens produced substantially higher aqueous concentrations than the known minimum inhibitory concentration for most bacteria. Moxifloxacin concentration in aqueous humour with multiple instillations is significantly higher than single instillation (P<0.05), which is adequate to cover ciprofloxacin-resistant gram-negative bacteria. Repeated topical moxifloxacin administration achieved significantly higher aqueous humour concentrations than single administration.

Nanotechnology as a therapeutic tool to combat microbial resistance.

Use of nanoparticles is among the most promising strategies to overcome microbial drug resistance. This review article consists of three parts. The first part discusses the epidemiology of microbial drug resistance. The second part describes mechanisms of drug resistance used by microbes. The third part explains how nanoparticles can overcome this resistance, including the following: Nitric oxide-releasing nanoparticles (NO NPs), chitosan-containing nanoparticles (chitosan NPs), and metal-containing nanoparticles all use multiple mechanisms simultaneously to combat microbes, thereby making development of resistance to these nanoparticles unlikely. Packaging multiple antimicrobial agents within the same nanoparticle also makes development of resistance unlikely. Nanoparticles can overcome existing drug resistance mechanisms, including decreased uptake and increased efflux of drug from the microbial cell, biofilm formation, and intracellular bacteria. Finally, nanoparticles can target antimicrobial agents to the site of infection, so that higher doses of drug are given at the infected site, thereby overcoming resistance.

Unraveling a mechanism of honey antibacterial action: polyphenol/H2O2-induced oxidative effect on bacterial cell growth and on DNA degradation.

Several compounds with antibacterial activities were identified in honey however, a mechanism by which they lead to bacterial growth inhibition and bacterial death remains still unknown. We recently found that honeys possess DNA degrading activity mediated by honey hydrogen peroxide and an unknown honey component(s). Here we provide evidence that active honeys (MIC90 of 6.25-12.5% v/v) possessed significantly higher levels of phenolics (p<0.02) of higher radical scavenging activities (p<0.005) than honeys of average activity. Removal of H2O2 by catalase eliminated bacteriostatic activities caused by both phenolics and H2O2 suggesting that the growth inhibition resulted from the coupling chemistry between these compounds. Both phenolics and H2O2 were involved in DNA degradation by honeys. Treatment of plasmid DNA with H2O2 alone did not affect the DNA integrity but H2O2 removal from honey by catalase prevented DNA degradation. Polyphenols extracted from honeys degraded plasmid DNA in the presence of H2O2 and Cu(II) in the Fenton-type reaction. The extent of DNA degradation was inversely related to the polyphenol concentration in this system as well as in honeys. At low content, honey polyphenols exerted pro-oxidant activity damaging to DNA. In conclusion, honey phenolics with pro-oxidant activities were necessary intermediates that conferred oxidative action of H2O2. Phenolic/H2O2-induced oxidative stress constituted the mechanism of honey bacteriostatic and DNA damaging activities.