Multiresistant Clinical Isolates Research Articles

Antibiofilm Activity of Curcumin and Piperine and Their Synergistic Effects with Antifungals against Candida albicans Clinical Isolates.

Candidiasis is the common name for diseases caused by yeast of the genus Candida. Candida albicans is one of the most implicated species in superficial and invasive candidiasis. Antifungals, polyenes, and azoles have been used to treat candidiasis. However, due to the development of antifungal resistance, research of natural substances with potential antifungal effects at low concentrations or combined is also a possibility. The broth microdilution method was used to evaluate the antifungal activity. The biofilm formation was assessed using the microtiter plate method. The antibiofilm activities were assessed using micro plaque tetrazolium salt assay (MTT). The combination effect of antifungal with natural substances was made using the checkerboard method. Among our isolates, clotrimazole was the most resistant, but amphotericin B was the most effective antifungal. The biofilm was formed by all isolates of C. albicans. Curcumin and piperine displayed antibiofilm activity with minimum biofilm inhibitory concentration (MBIC) and minimum eradicating concentration (MBEC) ranging from 64 to 1024 μg/mL and 256 to 2048 μg/mL. In combination, piperine presented double synergistic effects compared to curcumin with all antifungals tested. Curcumin shows more synergistic effect when combined with polyenes than with azoles. However, piperine shows a more synergistic effect when combined with azoles compared to polyenes. C. albicans was susceptible to curcumin and piperine both on planktonic cells and biofilm. The combination of curcumin and piperine with antifungals has shown synergistic effects against multiresistant clinical isolates of Candida albicans representing an alternative drug research for the treatment of clinical candidiasis.

Co-production of blaNDM-1 and blaOXA-23 in multiresistant clinical isolates of Acinetobacter baumannii from Brazil

Acinetobacter baumannii is a common and dangerous non-fermenting gram-negative bacillus due to its various and increasing resistance mechanisms. Therefore, the objective of the work was to describe four NDM-1 -producing Acinetobacter isolates from Natal, northeastern Brazil. These four isolates were identified as Acinetobacter baumannii by MALDI-TOF. All isolates were characterized by antimicrobial susceptibility testing, with resistance to all β-lactams including carbapenems and positive for the synergy test with Ethylenediaminetetraacetic acid. PCR analysis was also performed for blaNDM-1, blaVIM-1, blaIMP-1, blaOXA-23, blaOXA-24/40, blaOXA-51, blaOXA-58, and blaOXA-143 genes and showed positive for the blaNDM-1 and blaOXA-23 genes in all isolates and this result was confirmed by sequencing. This is the first case of Acinetobacter baumannii strains harboring blaNDM-1 gene isolated from northeastern Brazil. This description emphasizes the need for new prevention and control strategies of the dissemination of NDM-1-producing Acinetobacter, which are associated with a high mortality rate.

Combination of Six Individual Derivatives of the Pom-1 Antibiofilm Peptide Doubles Their Efficacy against Invasive and Multi-Resistant Clinical Isolates of the Pathogenic Yeast Candida albicans

In previous studies, derivatives of the peptide Pom-1, which was originally extracted from the freshwater mollusk Pomacea poeyana, showed an exceptional ability to specifically inhibit biofilm formation of the laboratory strain ATCC 90028 as a model strain of the pathogenic yeast Candida albicans. In follow-up, here, we demonstrate that the derivatives Pom-1A to Pom-1F are also active against biofilms of invasive clinical C. albicans isolates, including strains resistant against fluconazole and/or amphotericin B. However, efficacy varied strongly between the isolates, as indicated by large deviations in the experiments. This lack of robustness could be efficiently bypassed by using mixtures of all peptides. These mixed peptide preparations were active against biofilm formation of all the isolates with uniform efficacies, and the total peptide concentration could be halved compared to the original MIC of the individual peptides (2.5 µg/mL). Moreover, mixing the individual peptides restored the antifungal effect of fluconazole against fluconazole-resistant isolates even at 50% of the standard therapeutic concentration. Without having elucidated the reason for these synergistic effects of the peptides yet, both the gain of efficacy and the considerable increase in efficiency by combining the peptides indicate that Pom-1 and its derivatives in suitable formulations may play an important role as new antibiofilm antimycotics in the fight against invasive clinical infections with (multi-) resistant C. albicans.

Synergistic activity and immunomodulatory potential of levofloxacin and Synoeca‐MP peptide against multi-resistant strains of Klebsiella pneumoniae

The purpose of this article is to study the isolated and combined effect of the peptides Synoeca-MP and IDR-1018 against multi-resistant clinical isolates of K. pneumoniae (Kp2177569 - LACEN) in vitro. The bactericidal activity of the peptide Synoeca-MP in combination with three different classes of commercial antimicrobials and its immunomodulatory potential was also evaluated. Synoeca-MP showed better antimicrobial activity than IDR-1018 and presented synergistic action combined with levofloxacin. Therefore, Synoeca-MP and levofloxacin, and the combination of both, were used in subsequent analyses. In the presence of heat-killed antigens, cellular viability and TNF-α levels was maintained, the production of NO increased and a reduction in IL-10 production was observed. The synergistic antibacterial effect between Synoeca-MP and levofloxacin was effective against multidrug-resistant strains of K. pneumoniae. The association of Synoeca-MP and levofloxacin may present a low modulating action of pro and anti-inflammatory mediators, based on these results.

Structural and Functional Insights Into Skl and Pal Endolysins, Two Cysteine-Amidases With Anti-pneumococcal Activity. Dithiothreitol (DTT) Effect on Lytic Activity.

We have structurally and functionally characterized Skl and Pal endolysins, the latter being the first endolysin shown to kill effectively Streptococcus pneumoniae, a leading cause of deathly diseases. We have proved that Skl and Pal are cysteine-amidases whose catalytic domains, from CHAP and Amidase_5 families, respectively, share an α3β6-fold with papain-like topology. Catalytic triads are identified (for the first time in Amidase_5 family), and residues relevant for substrate binding and catalysis inferred from in silico models, including a calcium-binding site accounting for Skl dependence on this cation for activity. Both endolysins contain a choline-binding domain (CBD) with a β-solenoid fold (homology modeled) and six conserved choline-binding loci whose saturation induced dimerization. Remarkably, Pal and Skl dimers display a common overall architecture, preserved in choline-bound dimers of pneumococcal lysins with other catalytic domains and bond specificities, as disclosed using small angle X-ray scattering (SAXS). Additionally, Skl is proved to be an efficient anti-pneumococcal agent that kills multi-resistant strains and clinical emergent-serotype isolates. Interestingly, Skl and Pal time-courses of pneumococcal lysis were sigmoidal, which might denote a limited access of both endolysins to target bonds at first stages of lysis. Furthermore, their DTT-mediated activation, of relevance for other cysteine-peptidases, cannot be solely ascribed to reversal of catalytic-cysteine oxidation.

Amphipathic Barbiturates as Mimics of Antimicrobial Peptides and the Marine Natural Products Eusynstyelamides with Activity against Multi-resistant Clinical Isolates.

We report a series of synthetic cationic amphipathic barbiturates inspired by the pharmacophore model of small antimicrobial peptides (AMPs) and the marine antimicrobials eusynstyelamides. These N,N'-dialkylated-5,5-disubstituted barbiturates consist of an achiral barbiturate scaffold with two cationic groups and two lipophilic side chains. Minimum inhibitory concentrations of 2-8 μg/mL were achieved against 30 multi-resistant clinical isolates of Gram-positive and Gram-negative bacteria, including isolates with extended spectrum β-lactamase-carbapenemase production. The guanidine barbiturate 7e (3,5-di-Br) demonstrated promising in vivo antibiotic efficacy in mice infected with clinical isolates of Escherichia coli and Klebsiella pneumoniae using a neutropenic peritonitis model. Mode of action studies showed a strong membrane disrupting effect and was supported by nuclear magnetic resonance and molecular dynamics simulations. The results express how the pharmacophore model of small AMPs and the structure of the marine eusynstyelamides can be used to design highly potent lead peptidomimetics against multi-resistant bacteria.

N-chlorotaurine, a potent weapon against multiresistant bacteria.

Aims N‐chlorotaurine (NCT) is a body‐own mild oxidizing antiseptic that can be applied topically as a well‐tolerated anti‐infective at many body sites. The objective of this study was to demonstrate its activity against representative nosocomial multidrug‐resistant bacteria.Methods and ResultsThe bactericidal activity of NCT was tested in quantitative killing assays against a panel of multiresistant Gram‐positive and Gram‐negative clinical isolates. N‐chlorotaurine (1%, 55 mmol l−1) reduced the number of CFU of strains of methicillin‐resistant Staphylococcus aureus, linezolid‐resistant Staphylococcus epidermidis, vancomycin‐resistant, and linezolid‐ and vancomycin‐resistant Enterococcus faecium, 3MRGN and 4MRGN Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii and Klebsiella pneumoniae by at least 2 log10 steps after 15 min and completely or nearly to the detection limit after 30 min at pH 7·1 and 37°C.ConclusionThe activity of NCT against these clinical isolates is similar to that against non‐resistant ATCC strains and therefore not influenced by antibiotic resistance. This can be explained by the oxidizing and chlorinating mechanism of action of NCT, which leads to an attack of multiple targets in the microorganisms.Significance and Impact of the StudyThe bactericidal spectrum of NCT is not restricted by resistance against antibiotics. Therefore, it can be used against resistant strains, too.

Мониторинг ведущей грамотрицательной микрофлоры и антибиотикорезистентности при остеомиелите

Introduction There is an urgent need for a surveillance system of regular monitoring of specific bacteria inducing various types of osteomyelitis to identify resistant isolates and optimize the use of antibiotics. Objective: monitoring of specific gram-negative bacteria and analysis of the antibiotic resistance of the strains isolated from osteomyelitis patients over a three-year period. Results and discussion P. aeruginosa was the first most common pathogen among gram-negative microorganisms isolated from the patients between 2017 and 2019. Prevalence of the isolates identified in 2019 decreased by 9.6 % as compared to 2017. Next frequently encountered clinical isolates were Enterobacter sp., Acinetobacter sp., Klebsiella sp. There was a twofold increase in K. pneumoniae strains isolated in 2019. Analysis of antibiotic susceptibility testing data revealed multiresistance of the Acinetobacter sp. strains in 2019 despite the total decrease in resistant isolates in 2017 and 2018. Among non-fermenting gram-negative rods, the species being resistant to imipenem were shown to increase by 5.4 times. Overall antibiotic resistance was on rise. Increased antimicrobial resistance to beta-lactam antibiotics also combined with BLaC inhibitors was observed in Enterobacteriaceae population. Meropenem was found to be effective against most bacteria with growing drug resistance observed as compared with recent years. The antibiotic resistance profiles of Klebsiella sp. strains appeared to be high at antimicrobial testing. Conclusion Diverse bacterial morphology of gram-negative species and increasing proportion of drug-resistant strains isolated in osteomyelitis cases have necessitated regular monitoring of multiresistant clinical isolates for adjustment of empirical antibiotic therapies.

The Synergistic Effect of Combining Some Antibiotics with Powder of Silver NPs, Copper NPs and Titanium NPs

Routine antibiotics susceptibility testing still relies on standardized cultivation-based analyses, includingmeasurement of inhibition zones in conventional agar diffusion tests.The sensitivity of the bacterial isolatescausing Clinical isolatest results showed that all the isolates subjected to this study showed varying resistanceto the antibiotics used. The effect of nanoparticles combined with different antibiotics was investigatedagainst multi resistant Clinical isolates bacteria using disk diffusion method. The inhibition zones diameter(mm) around various antibiotic discs (Trimethoprim, sulphamethaxazol, Azithromycin, Aztreonam,Ceftriaxone, Gentamycin, Tetracyclin, Nitrofurantoin) with/without nanopatrticles against isolates weremeasured. The antibacterial activities of some antibiotics have been increased in the presence of nanoparticleagainst all isolates while others showed no effect. The highest antibacterial activities of antibiotics combinedwith nanoparticle were observed against E.coli isolates for Gentamycin, Trimethoprim-sulphamethaxazol,Azithromycin ,Tetracyclin ,Nitrofurantoin. while A. baumanii appeared sensitive to Gentamycin,Trimethoprim sulphamethaxazol and Azithromycin compared with antibiotics alone. K.pneumoniae becamesensitive to Gentamycin, Tetracyclin and P.aeruginosa to Ampicillin,Trimethoprim sulphamethaxazol,Azithromycin and Gentamycin. nanoparticle showed no effect on antibacterial activities of all antibioticsexcept Gentamycin against gram positive bacteria S.aureus.

Shorter Antibacterial Peptide Having High Selectivity for E. coli Membranes and Low Potential for Inducing Resistance.

Antimicrobial peptides (AMPs) have been recognised as a significant therapeutic option for mitigating resistant microbial infections. It has been found recently that Plasmodium falciparum-derived, 20 residue long, peptide 35409 had antibacterial and haemolytic activity, making it an AMP having reduced selectivity, and suggesting that it should be studied more extensively for obtaining new AMPs having activity solely targeting the bacterial membrane. Peptide 35409 was thus used as template for producing short synthetic peptides (<20 residues long) and evaluating their biological activity and relevant physicochemical characteristics for therapeutic use. Four of the sixteen short peptides evaluated here had activity against E. coli without any associated haemolytic effects. The 35409-1 derivative (17 residues long) had the best therapeutic characteristics as it had high selectivity for bacterial cells, stability in the presence of human sera, activity against E. coli multiresistant clinical isolates and was shorter than the original sequence. It had a powerful membranolytic effect and low potential for inducing resistance in bacteria. This peptide’s characteristics highlighted its potential as an alternative for combating infection caused by E. coli multiresistant bacteria and/or for designing new AMPs.

Discovery of a novel integron-borne aminoglycoside resistance gene present in clinical pathogens by screening environmental bacterial communities

BackgroundNew antibiotic resistance determinants are generally discovered too late, long after they have irreversibly emerged in pathogens and spread widely. Early discovery of resistance genes, before or soon after their transfer to pathogens could allow more effective measures to monitor and reduce spread, and facilitate genetics-based diagnostics.ResultsWe modified a functional metagenomics approach followed by in silico filtering of known resistance genes to discover novel, mobilised resistance genes in class 1 integrons in wastewater-impacted environments. We identified an integron-borne gene cassette encoding a protein that conveys high-level resistance against aminoglycosides with a garosamine moiety when expressed in E. coli. The gene is named gar (garosamine-specific aminoglycoside resistance) after its specificity. It contains none of the functional domains of known aminoglycoside modifying enzymes, but bears characteristics of a kinase. By searching public databases, we found that the gene occurs in three sequenced, multi-resistant clinical isolates (two Pseudomonas aeruginosa and one Luteimonas sp.) from Italy and China, respectively, as well as in two food-borne Salmonella enterica isolates from the USA. In all cases, gar has escaped discovery until now.ConclusionTo the best of our knowledge, this is the first time a novel resistance gene, present in clinical isolates, has been discovered by exploring the environmental microbiome. The gar gene has spread horizontally to different species on at least three continents, further limiting treatment options for bacterial infections. Its specificity to garosamine-containing aminoglycosides may reduce the usefulness of the newest semisynthetic aminoglycoside plazomicin, which is designed to avoid common aminoglycoside resistance mechanisms. Since the gene appears to be not yet common in the clinics, the data presented here enables early surveillance and maybe even mitigation of its spread.

Antimicrobial activity of amphipathic α,α-disubstituted β-amino amide derivatives against ESBL – CARBA producing multi-resistant bacteria; effect of halogenation, lipophilicity and cationic character

The rapid emergence and spread of multi-resistant bacteria have created an urgent need for new antimicrobial agents. We report here a series of amphipathic α,α-disubstituted β-amino amide derivatives with activity against 30 multi-resistant clinical isolates of Gram-positive and Gram-negative bacteria, including isolates with extended spectrum β-lactamase – carbapenemase (ESBL-CARBA) production. A variety of halogenated aromatic side-chains were investigated to improve antimicrobial potency and minimize formation of Phase I metabolites. Net positive charge and cationic character of the derivatives had an important effect on toxicity against human cell lines. The most potent and selective derivative was the diguanidine derivative 4e with 3,5-di-brominated benzylic side-chains. Derivative 4e displayed minimum inhibitory concentrations (MIC) of 0.25–8 μg/mL against Gram-positive and Gram-negative reference strains, and 2–32 μg/mL against multi-resistant clinical isolates. Derivative 4e showed also low toxicity against human red blood cells (EC50 > 200 μg/mL), human hepatocyte carcinoma cells (HepG2: EC50 > 64 μg/mL), and human lung fibroblast cells (MRC-5: EC50 > 64 μg/mL). The broad-spectrum antimicrobial activity and low toxicity of diguanylated derivatives such as 4e make them attractive as lead compounds for development of novel antimicrobial drugs.

Antibacterial activity of a Tribolium castaneum defensin in an in vitro infection model of Streptococcus pneumoniae

ABSTRACTStreptococcus pneumoniae (S. pneumoniae) is the most common bacterial cause of community-acquired pneumonia. Increasing rates of antibiotic-resistant S. pneumoniae strains impair therapy and necessitate alternative treatment options. In this study, we analysed insect-derived antimicrobial peptides (AMPs) for antibacterial effects on S. pneumoniae in a human in vitro infection model.AMP effects on bacterial growth were examined by colony forming unit (CFU)-assays, and growth curve measurements. Furthermore, cytotoxicity to primary human macrophages was detected by measuring lactate-dehydrogenase release to the supernatant. One AMP (Defensin 1) was tested in a model of primary human monocyte-derived macrophages infected with S. pneumoniae strain D39 and a multi-resistant clinical isolate. Inflammatory reactions were characterised by qPCR and multiplex-ELISA.In total, the antibacterial effects of 23 AMPs were characterized. Only Tribolium castaneum Defensin 1 showed significant antibacterial effects against S. pneumoniae strain D39 and a multi-resistant clinical isolate. During in vitro infection of primary human macrophages with S. pneumoniae D39, Defensin 1 displayed strong antibacterial effects, and consequently reduced bacteria-induced cytokine expression and release.In summary, Tribolium castaneum Defensin 1 showed profound antibacterial effectivity against Streptococcus pneumoniae D39 and a multi-resistant clinical isolate without unwanted cytotoxic or inflammatory side effects on human blood-derived macrophages.

Bactericidal synergism between antibiotics and phage endolysin Cpl-711 to kill multidrug-resistant pneumococcus.

Aim:To test the synergistic effect of Cpl-711 endolysin and antibiotics for antipneumococcal activity.Materials & methods:A combination of Cpl-711 and different antibiotics (amoxicillin, cefotaxime, levofloxacin and vancomycin) was tested in a checkerboard assay against several multidrug-resistant Streptococcus pneumoniae strains. Mouse and zebrafish models of pneumococcal sepsis were used to confirm the in vitro data.Results:The activity of Cpl-711 combined with amoxicillin or cefotaxime was synergistic in the bactericidal effect against a serotype 23F multiresistant clinical isolate of S. pneumoniae. Synergy between Cpl-711 and cefotaxime was validated using both mouse and zebrafish models.Conclusion:Combination of Cpl-711 and cefotaxime may help in the treatment of diseases caused by multiresistant pneumococcal strains.

An amphipathic cyclic tetrapeptide scaffold containing halogenated β2,2‐amino acids with activity against multiresistant bacteria

The present study describes the synthesis and biological studies of a small series of head-to-tail cyclic tetrapeptides of the general structure c(Lys-β2,2 -Xaa-Lys) containing one lipophilic β2,2 -amino acid and Lys, Gly, Ala, or Phe as the Xaa residue in the sequence. The peptides were investigated for antimicrobial activity against gram-positive and gram-negative reference strains and 30 multiresistant clinical isolates including strains with extended spectrum β-lactamase-carbapenemase (ESBL-CARBA) production. Toxicity was determined against human red blood cells. The most potent peptides showed high activity against the gram-positive clinical isolates with minimum inhibitory concentrations of 4-8μg/mL and low haemolytic activity. The combination of high antimicrobial activity and low toxicity shows that these cyclic tetrapeptides containing lipophilic β2,2 -amino acids form a valuable scaffold for designing novel antimicrobial agents.

A novel tool against multiresistant bacterial pathogens: lipopeptide modification of the natural antimicrobial peptide ranalexin for enhanced antimicrobial activity and improved pharmacokinetics

As evolutionarily optimised defence compounds, antimicrobial peptides (AMPs) represent a powerful tool against bacterial infections. Ranalexin, an AMP found in the skin of the American bullfrog (Rana catesbeiana), is primarily active against Gram-positive bacteria, with minimum inhibitory concentrations (MICs) of 8-16 mg/L, but shows weaker activity against Gram-negative bacteria (MICs > 64 mg/L). By substitution of six N-terminal amino acids by saturated fatty acids [decanoic acid (C10:0) to myristic acid (C14:0)], lipopeptide derivatives with enhanced antimicrobial activity were developed. The antimicrobial capacity of the peptides was tested against different bacterial strains, including multiresistant clinical isolates. C13C3lexin, the most potent derivative, showed MICs of 2-8 mg/L against Gram-positive bacteria and 2-16 mg/L against Gram-negative bacteria. In time-kill studies, it was clearly shown that ranalexin and the lipopeptide C13C3lexin function as concentration-dependent, fast-acting substances against different bacteria. Cell viability assays revealed that cytotoxicity towards human cells increases with the chain length of the attached fatty acid (IC50, 12.74-108.10 µg/mL). Furthermore, using positron emission tomography (PET) imaging, pharmacokinetic studies of 68Ga-labelled ranalexin and its derivatives were performed for the first time. Here it was demonstrated that ranalexin is rapidly cleared via the kidneys within 1 h post-injection. In contrast, the lipopeptide showed greatly extended circulation in the bloodstream and a shift from renal to hepatic accumulation characteristics. Therefore, the more favourable pharmacokinetics and enhanced antimicrobial activity clearly demonstrate the potential of the lipopeptide AMPs as novel ammunition against emerging multiresistant bacterial pathogens.

A novel synthetic peptide inspired on Lys49 phospholipase A2 from Crotalus oreganus abyssus snake venom active against multidrug-resistant clinical isolates

Currently, the evolving and complex mechanisms of bacterial resistance to conventional antibiotics are increasing, while alternative medicines are drying up, which urges the need to discover novel agents able to kill antibiotic-resistant bacteria. Lys49 phospholipase A2s (PLA2s) from snake venoms are multifunctional toxins able to induce a huge variety of therapeutic effects and consequently serve as templates for new drug leads. Hence, the present study was aimed at the synthesis of oligopeptides mimicking regions of the antibacterial Lys49 PLA2 toxin (CoaTx-II), recently isolated from Crotalus oreganus abyssus snake venom, to identify small peptides able to reproduce the therapeutic action of the toxin. Five peptides, representing major regions of interest within CoaTx-II, were synthesized and screened for their antibacterial properties. The 13-mer peptide pC-CoaTxII, corresponding to residues 115–129 of CoaTx-II, was able to reproduce the promising bactericidal effect of the toxin against multi-resistant clinical isolates. Peptide pC-CoaTxII is mainly composed by positively charged and hydrophobic amino acids, a typical trait in most antimicrobial peptides, and presented no defined secondary structure in aqueous environment. The physicochemical properties of pC-CoaTxII are favorable towards a strong interaction with anionic lipid membranes as those in bacteria. Additional in silico studies suggest formation of a water channel across the membrane upon peptide insertion, eventually leading to bacterial cell disruption and death. Overall, our findings confirm the valuable potential of snake venom toxins towards design and synthesis of novel antimicrobials, thus representing key insights towards development of alternative efficient antimicrobials to fight bacterial resistance to current antibiotics.

Antimycobacterial activity of veterinary antibiotics (Apramycin and Framycetin) against Mycobacterium abscessus: Implication for patients with cystic fibrosis.

Antimicrobial resistance has rendered certain species of Mycobacterium difficult to treat clinically, particularly the nontuberculous Mycobacterium and Mycobacterium abscessus. While veterinary medicine and human medicine share many classes of antibiotics, there are several antibiotics which are uniquely licensed to veterinary medicine but not human medicine. It was, therefore, the aim of this study to examine the action of eight veterinary antibiotics to a population of multi- and pan-resistant M. abscessus, isolated from the sputum of patients with cystic fibrosis (CF). Antibiotic susceptibility studies were performed on human clinical isolates of M. abscessus (n = 16), including 11 smooth isolates, 4 rough isolates, and 1 reference isolate (NCTC 13031), against the following 7 veterinary antibiotics (antibiotic class): apramycin (aminoglycoside), cefovecin (cephalosporin), ceftiofur (cephalosporin), framycetin (aminoglycoside), lincomycin (lincosamide), pirlimycin (lincosamide), and spectinomycin (aminocyclitol). M. abscessus isolates were sensitive (100%) to apramycin and framycetin but resistant (100%) to cefovecin, ceftiofur, lincomycin, pirlimycin, and spectinomycin. This study identified that the veterinary aminoglycosides, apramycin, and framycetin, have in vitro activity against multi-resistant clinical isolates of M. abscessus. Further studies should now compare the activity of these antibiotics against amikacin and the human aminoglycoside, advocated in the treatment of disease in CF patients, to determine if these novel antibiotics have a future role for the development in human medicine with such chronic disease patients.

Curcumin, an antibiotic resistance breaker against a multiresistant clinical isolate of Mycobacterium abscessus.

Curcumin, a phenolic compound extracted from Curcuma longa, exerts multiple pharmacological effects, including an antimicrobial action. Mycobacterium abscessus, an environmental, nontuberculous, rapidly growing mycobacterium, is an emerging human pathogen causing serious lung infections and one of the most difficult to treat, due to its multidrug resistance and biofilm-forming ability. We wanted to evaluate the antimicrobial and antivirulence activity of curcumin and its ability to synergize with antibiotics against a clinical M.abscessus strain (29904), isolated from the bronchoaspirate of a 66-year-old woman admitted to hospital for suspected tuberculosis. Curcumin [minimum inhibitory concentrations (MIC)=128mg/L] was synergic (fractional inhibitory concentration index ≤0.5) with amikacin, clarithromycin, ciprofloxacin, and linezolid, to which strain 29904 showed resistance/intermediate susceptibility. Curcumin at 1/8×MIC significantly reduced motility, whereas at 4×MIC, it completely inhibited 4- and 8-day mature biofilms. Synergistic combinations of curcumin and amikacin induced a general reduction in microbial aggregates and substantial loss in cell viability. Disruption of 4- and 8-day biofilms was the main effect detected when curcumin was the predominant compound. The present findings support previous evidence that curcumin is a potential antibiotic resistance breaker. Curcumin, either alone or combined with antibiotics, could provide a novel strategy to combat antibiotic resistance and virulence of M.abscessus.

Anti-biofilm activity of A22 ((S-3,4-dichlorobenzyl) isothiourea hydrochloride) against Pseudomonas aeruginosa: Influence on biofilm formation, motility and bioadhesion

Bacterial biofilms are involved in various medical infections and for this reason it is of great importance understanding adhesion mechanisms of involved microorganisms is essential to develop new strategies of prevention and control. Different approaches have been used for preventing biofilm related infections in health care settings, such as use of surface coatings agents in medical implants. In this context, is necessary to explore new compounds with anti-biofilm activity. Thus, this study evaluated for the first time the action of A22 against biofilms of Pseudomonas aeruginosa PAO1 strain and multi-resistant clinical isolates on biotic and abiotic surfaces. A22 acts as inhibitor of the MreB protein of the bacterial cell wall, causing the rods to change shape to the coccoid form. In this work, A22 at subinhibitory concentrations was able to prevent biofilm formation, and atomic force microscopy images showed that A22 was highly effective in inhibiting adhesion on polyethylene surfaces. Pseudomonas aeruginosa PAO1 exhibited a strong ability to adhere to HeLa cells, and A22 inhibited the aggregation after 4 h of exposure. Swarming and twitching motilities were significantly altered by A22 at subinhibitory concentrations. Thus, by changing the shape of the bacterial cell, many properties can be affected, such as motility, surface adhesion and biofilm formation. This work presents A22 as a promising novel antibacterial or surface coating agent of medical materials.