Development Of Bacterial Resistance Research Articles

Knowledge and practices of using antibiotics by patients with acute respiratory infections

Introduction. Excessive and inappropriate antibiotic (AB) consumption is associated with the development of antibiotic resistance (AR). Insufficient knowledge and irrational practices in the use of AB for acute respiratory infections (ARI) by patients contribute to the development of AR. Aim: to assess the knowledge and practices of AB use in patients with ARI in order to identify educational needs.
 Material and methods. The study included 393 patients with ARI. The standardized Happy Audit-2 questionnaire was applied. Complaints, the frequency of ARI occurrence, administered AB treatment, AB acquisition method, AB compliance, knowledge about AB, and AB resistance were assessed.
 Results. AB were used by 40.3% of the patients. Antibiotic treatment was recommended by a doctor in 87.6% of cases, taken independently by 5.6%, recommended by a pharmacist in 5.0%, and recommended by relatives in 1.9% of cases. AB were obtained without a prescription in 6.3% of cases, and household reserves were used in 20.8% of cases. Patients were compliant with antibiotic treatment in 89.7% of cases but 10.3% of respondents did not adhere to the prescribed duration/dosage. 26.7% of respondents believed that AB were effective against viral infections, and 13.8% believed they were harmless to the human body. In 24.1% of cases, respondents were unaware of the adverse effects of AB, including the development of bacterial resistance in 29.7% of cases.
 Conclusions. Insufficient knowledge about the effects of antibiotics and irrational usage practices, including self-medication, emphasize the need for strengthening awareness and education measures for potential antibiotic users.

Impact of CRAMP-34 on Pseudomonas aeruginosa biofilms and extracellular metabolites.

Biofilm is a structured community of bacteria encased within a self-produced extracellular matrix. When bacteria form biofilms, they undergo a phenotypic shift that enhances their resistance to antimicrobial agents. Consequently, inducing the transition of biofilm bacteria to the planktonic state may offer a viable approach for addressing infections associated with biofilms. Our previous study has shown that the mouse antimicrobial peptide CRAMP-34 can disperse Pseudomonas aeruginosa (P. aeruginosa) biofilm, and the potential mechanism of CRAMP-34 eradicate P. aeruginosa biofilms was also investigated by combined omics. However, changes in bacterial extracellular metabolism have not been identified. To further explore the mechanism by which CRAMP-34 disperses biofilm, this study analyzed its effects on the extracellular metabolites of biofilm cells via metabolomics. The results demonstrated that a total of 258 significantly different metabolites were detected in the untargeted metabolomics, of which 73 were downregulated and 185 were upregulated. Pathway enrichment analysis of differential metabolites revealed that metabolic pathways are mainly related to the biosynthesis and metabolism of amino acids, and it also suggested that CRAMP-34 may alter the sensitivity of biofilm bacteria to antibiotics. Subsequently, it was confirmed that the combination of CRAMP-34 with vancomycin and colistin had a synergistic effect on dispersed cells. These results, along with our previous findings, suggest that CRAMP-34 may promote the transition of PAO1 bacteria from the biofilm state to the planktonic state by upregulating the extracellular glutamate and succinate metabolism and eventually leading to the dispersal of biofilm. In addition, increased extracellular metabolites of myoinositol, palmitic acid and oleic acid may enhance the susceptibility of the dispersed bacteria to the antibiotics colistin and vancomycin. CRAMP-34 also delayed the development of bacterial resistance to colistin and ciprofloxacin. These results suggest the promising development of CRAMP-34 in combination with antibiotics as a potential candidate to provide a novel therapeutic approach for the prevention and treatment of biofilm-associated infections.

A dual mechanism with H2S inhibition and membrane damage of morusin from Morus alba Linn. against MDR-MRSA

It’s urgent to discover new antibiotics along with the increasing emergence and dissemination of multidrug resistant (MDR) bacterial pathogens. In the present investigation, morusin exhibited rapid bactericidal activity against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE) by targeting the phospholipid of bacterial inner membrane, increasing membrane rigidity and disrupting bacterial homeostasis together with the membrane permeability, which caused fundamental metabolic disorders. Furthermore, morusin can also accumulate ROS, suppress H2S production, and aggravate oxidative damage in bacteria. Importantly, morusin also inhibited the spread of wounds and reduced the bacterial burden in the mouse model of skin infection caused by MRSA. It’s a chance to meet the challenge of existing antibiotic resistance and avoid the development of bacterial resistance, given the multiple targets of morusin.

The Anti-Virulence Activities of the Antihypertensive Drug Propranolol in Light of Its Anti-Quorum Sensing Effects against Pseudomonas aeruginosa and Serratia marcescens.

The development of bacterial resistance is an increasing global concern that requires discovering new antibacterial agents and strategies. Bacterial quorum sensing (QS) systems play important roles in controlling bacterial virulence, and their targeting could lead to diminishing bacterial pathogenesis. In this context, targeting QS systems without significant influence on bacterial growth is assumed as a promising strategy to overcome resistance development. This study aimed at evaluating the anti-QS and anti-virulence activities of the β-adrenoreceptor antagonist propranolol at sub-minimal inhibitory concentrations (sub-MIC) against two Gram-negative bacterial models Pseudomonas aeruginosa and Serratia marcescens. The effect of propranolol on the expression of QS-encoding genes was evaluated. Additionally, the affinity of propranolol to QS receptors was virtually attested. The influence of propranolol at sub-MIC on biofilm formation, motility, and production of virulent factors was conducted. The outcomes of the propranolol combination with different antibiotics were assessed. Finally, the in vivo protection assay in mice was performed to assess propranolol's effect on lessening the bacterial pathogenesis. The current findings emphasized the significant ability of propranolol at sub-MIC to reduce the formation of biofilms, motility, and production of virulence factors. In addition, propranolol at sub-MIC decreased the capacity of tested bacteria to induce pathogenesis in mice. Furthermore, propranolol significantly downregulated the QS-encoding genes and showed significant affinity to QS receptors. Finally, propranolol at sub-MIC synergistically decreased the MICs of different antibiotics against tested bacteria. In conclusion, propranolol might serve as a plausible adjuvant therapy with antibiotics for the treatment of serious bacterial infections after further pharmacological and pharmaceutical studies.

2814. The Impact of Novel Antibiotics on Curbing Antimicrobial Resistance and Infection Spread a Dynamic Modeling Study

Abstract Background Antimicrobial resistance (AMR) is a growing global concern. Novel antibiotics (ABXs) target resistant pathogens causing gram-negative (GN) bacterial infections. This study assesses the broader impact of novel ABX use on the US healthcare system, including preventing the onward spread of infections and reducing the level of high-threat AMR. Methods A stochastic dynamic transmission model was developed to simulate the interaction between infection transmissibility, bacterial resistance development, and multiple lines of ABX. Included infection types were intra-abdominal infections, urinary tract infections and hospital-acquired /ventilator-associated pneumonia. Included AMR types were carbapenem-resistant and ESBL-producing enterobacterales (CRE and ESBLE), and multi-drug resistant pseudomonas aeruginosa (MDRP). The model has compartments for non-infected, colonized, or infected patients, with a sensitive or resistant strain in a US hospital. Infected patients receive antibiotics resulting in infection clearance or use of an alternate ABX class. Sensitive pathogens gain resistance due to drug pressure or conversely could lose resistance. Ordinary differential equations were used to estimate compartment membership. The primary data source was the Premier healthcare database (172 hospitals, 2016-2020). The model examined the potential reduction in AMR levels and avoided hospital infections, by comparing two scenarios: targeted use of novel ABX for specific AMR types and the status quo with limited novel ABX use. Results When even novel ABXs are reserved as the 2nd or 3rd line of treatment, compared to the status quo, it could result in a ∼1.5% reduction in hospital infections and a change in CRE, ESBLE, and MDRP by −2.5%, −3.6%, and −2.1%, respectively, on an annual basis. The cumulative impact of novel ABXs over 5-10 years was much larger but subject to higher prediction uncertainty. The primary driver of results are infection clearance rates, risk of resistance gain on various ABX, and uptake of novel ABX. Conclusion Targeted use of novel ABXs for resistant pathogens could lead to improved, earlier infection clearance with most types of infections and pathogens, thereby reducing infection transmission and rise of high-threat AMR in the long-term. Disclosures Apoorva Ambavane, PhD, Merck & Co., Inc: Grant/Research Support Michal Litkiewicz, MSc, Merck & Co., Inc: Grant/Research Support Aditya Sardesai, MSc, CSL Seqirus: Advisor/Consultant|Merck & Co. Inc: Grant/Research Support|Merck & Co. Inc: Grant/Research Support Diana Teloian, MA, Merck & Co., Inc: Grant/Research Support Engels N. Obi, PhD, Merck & Co Inc: Employee Joe Yang, PhD, Merck & Co., Inc: Employee|Merck & Co., Inc: Stocks/Bonds

The Outcome of Antibiotic Overuse before and during the COVID-19 Pandemic in a Tertiary Care Hospital in Oman.

Antimicrobial resistance (AMR), a serious global public health challenge, may have accelerated development during the COVID-19 pandemic because antibiotics were prescribed for COVID-19. This study aimed to assess antibiotics use before and during the pandemic and correlate the results with the rate of resistant microorganisms detected in hospitalized patients during the study period. This single-center study looked retrospectively at four years of data (2018-2021) from Royal Hospital, Muscat, which is the biggest hospital in Oman with approximately 60,000 hospital admissions yearly. The consumption rate of ceftriaxone, piperacillin tazobactam, meropenem, and vancomycin was presented as the antibiotic consumption index, the ratio of defined daily dose (DDD) per 100 bed days. Analyses were performed using the nonparametric test for trend across the study period. Correlation between antibiotic consumption indexes and the isolated microorganisms in the four-year study period was performed using Spearman's rank correlation coefficient. We compared data from the pre-COVID-19 to the COVID-19 period. Though more patients were admitted pre-COVID-19 (132,828 versus 119,191 during COVID-19), more antibiotics were consumed during the pandemic (7350 versus 7915); vancomycin and ceftriaxone had higher consumption during than before the pandemic (p-values 0.001 and 0.036, respectively). Vancomycin-resistant Enterococcus (VRE) and Candida auris were detected more during the COVID-19 period with p-values of 0.026 and 0.004, respectively. Carbapenem-resistant Enterobacterales (CRE), vancomycin-resistant Enterococcus spp., and C. auris were detected more often during the pandemic with p-values of 0.011, 0.002, and 0.03, respectively. Significant positive correlations between antibiotic consumption and drug-resistant isolates were noted. This study confirms that the overuse of antibiotics triggers the development of bacterial resistance; our results emphasize the importance of antibiotic control.

Knowledge and Awareness of the Therapeutic Benefits and Precautions of Natural Honey Consumption among students

Scientists encourage using honey as an alternative therapy for some infectious disease due to the development of bacterial resistance against antibiotics and decreased some of their effectiveness in clinical practice. Different types of honey have different actions and activities. The aim of this study was to evaluate the knowledge of the therapeutic benefits and precautions of using natural honey among medical sciences students. This is a cross-sectional study that used through a survey questioner which was created using Google form tool and was distributed to undergraduate medicine (MD) and pharmacy students (B. Pharm) in Muscat, Oman. The results of this study found that Omani honey was mostly preferred by 53.4% of the participants. Although honey rich in health benefits it was concluded that 58.2% of participants never advised by a doctor to use honey for a certain medical condition. It was found that 71.9% not aware about anti-inflammatory effects of honey to reduce eye inflammation. While more than 70% know about efficiently of cosmetics uses of honey. Moreover, almost half of the participants (49%) seem to be unaware of the suitable age to start taking honey which may be fatal to infants. Therefore, parents should be warned not to give honey to their new-born to prevent them from the botulism toxicity. The awareness about the benefits of honey is accepted in some consideration while insufficient in the awareness precautions of using honey with children under the age of 12 months which may be fatal.

Comprehensive study on the potential environmental risk of temporal antibiotic usage through wastewater discharges

Antibiotic residues can reach aquatic ecosystems through urban wastewater discharges, posing an ecotoxicological risk for aquatic organisms and favoring the development of bacterial resistance. To assess the emission rate and hazardousness of these compounds, it is important to carry out periodic chemical monitoring campaigns that provide information regarding the actual performance of wastewater treatment plants (WWTPs) and the potential impact of the treated wastewater in the aquatic environment. In this study, 18 of the most widely consumed antibiotics in Spain were determined by liquid chromatography-tandem mass spectrometry in both influent (IWW) and effluent wastewater (EWW) samples collected over four seasons along 2021–2022. Eleven antibiotics were detected in EWW with azithromycin, ciprofloxacin and levofloxacin showing the highest concentration levels (around 2 μg L−1 of azithromycin and 0.4 μg L−1 of quinolone compounds). Data showed that only 4 out of the 11 compounds were removed by more than 50 % in the WWTP, with sulfamethoxazole standing out with an average removal efficiency >80 %. The risk that treated water could pose to the aquatic environment was also assessed, with 6 compounds indicating a potential environmental risk by exceeding established ecotoxicological and resistance thresholds. Based on the risk assessment, the WWTP removal efficiency required to reduce such risk for antibiotics was estimated. In addition, pooled wastewater samples were screened by LC coupled to high resolution mass spectrometry with ion mobility separation, searching for metabolites and transformation products of the antibiotics investigated to widen future research. Studies like this are crucial to map the impact of antibiotic pollution and to provide the basis for designing water quality and risk prevention monitoring programs.

Repeat-exposure in vitro protocol to assess the risk of antimicrobial resistance (AMR) development from use of personal care products: Case study using an antibacterial liquid handwash

The global crisis we are facing with regard to antibiotic resistance has been largely attributed to the overuse and misuse of antibiotics in healthcare and agriculture. However, there is also growing global concern about cross-resistance between biocides and antibiotics. This has made clear the need for more research in this area along with easy-to-perform, but realistic, methods to characterise the potential risk associated with cross-resistance to antibiotics due to biocide use. The primary aim of this work was to develop a repeat-exposure method for predicting bacterial resistance to microbicides, including their cross-resistance to antibiotics. Realism is incorporated in the presented protocol through the use of relevant concentrations and contact times, validated neutralisers, appropriate test organisms and repeat-exposures. The protocol can be applied to formulated microbicides, as shown in the liquid handwash case study presented here. Five bacterial strains were included in the study: Staphylococcus aureus ATCC 6538, Pseudomonas aeruginosa ATCC 15442, Staphylococcus epidermidis ATCC 14990, Escherichia coli ATCC 10536 and Enterococcus hirae ATCC 10541. The protocol parameters used in the case study reflected a worst-case exposure scenario (in terms of contact time and concentration). The results demonstrated that repeated exposure to the liquid handwash would not be expected to lead to development of bacterial resistance or cross-resistance to antibiotics. It is envisaged that this protocol could be used by manufacturers of microbicidal formulations to assess whether repeated use of the test products would contribute to bacterial resistance development or cross-resistance to antibiotics.

Hype or healing: unveiling the truth behind manuka honey?

Aim of the study was to find out the effectiveness of Manuka honey (MH) in various applications to treat drug resistant ENT infections as an adjuvant therapy. The antibacterial properties and antioxidant properties of MH is attributed to the presence of high concentration of methylglyoxal (MGO) and flavonoid, phenolic compounds respectively. Unique manuka factor (UMF) rating is a scale used to determine the antibacterial potency of MH which directly correlates to the MGO and phenolic components in MH. Various cases of nasal crusting in post operative period with multidrug resistant bacterial cultures were included in the study. Each case was started on thorough nasal douching as well as manuka honey topical application. Regular and meticulous follow up was done for each case via endoscopic examination and medical record was kept of any potential side effects. Perichondritis of pinna was included in the study with potentially unique application the above-mentioned condition. MH is a natural antibiotic with low toxicity profile, allowing for prolonged use when necessary. Moreover, the development of bacterial resistance to honey has yet to be reported, making it an attractive alternative or adjunctive therapy, particularly in cases where antimicrobial resistance is a concern. While Manuka honey's antimicrobial properties have been observed in various studies, translating this knowledge into clinical practice requires rigorous investigation to ensure its effectiveness, safety, and proper integration with conventional antibiotic therapies.

Group B streptococcal and premature births: a narrativo review

Introduction: Premature births are those that occur before 37 weeks of gestational age. A clinical condition that remains problematic for obstetrics, mainly because of the high neonatal mortality it causes. Although most children survive, they are at risk of increased damage to neurological development and respiratory and gastrointestinal complications. Microorganisms, such as Streptococcal agalactiae (Group B Streptococcal - GBS), have been associated with prematurity. This comprehensive review aims to present data on the association between GBS and prematurity. Methods: Using the keywords Group B Streptococcal, prematurity, and Intrapartum antibiotic therapy, articles on the following databases were searched: PubMed, the Cochrane database of Systematic Reviews, the Cochrane Central Register of Controlled Trials, and Lilacs/Scielo. The most relevant works were considered those published on this theme from January 1, 1973 to December 31, 2021 available on the databases consulted. Results: Premature rupture of ovular membranes (PROM) occurs in 1% to 3% of pregnancies, being an important cause of perinatal morbidity and mortality and being associated with 30-40% of premature births. Infection caused by group B streptococcal, has been indicated as an important risk factor of premature birth, especially in patients with premature amniorrhexis of the ovular membranes. Discussion: Preventive prophylaxis measures for pregnant women, such as intrapartum medication, to reduce the vertical transmission of invasive GBS diseases resulted in a significant decrease in early newborn disease by the pathogen. In addition, penicillin was and remains as the antimicrobial of choice due to the fact that it has a narrower microbicidal spectrum than the ampicillin, and so it reduces the likelihood of the development of bacterial resistance. Conclusion: GBS has been associated with increased risk of preterm delivery due to premature rupture of the membrane. Also, antibiotic therapy for vaginal infection by bacteria reduced preterm birth with low weight in some populations.

Eobania vermiculata whole-body muscle extract-loaded chitosan nanoparticles enhanced skin regeneration and decreased pro-inflammatory cytokines in vivo

BackgroundUsually, wounds recover in four to six weeks. Wounds that take longer time than this to heal are referred to as chronic wounds. Impaired healing can be caused by several circumstances like hypoxia, microbial colonization, deficiency of blood flow, reperfusion damage, abnormal cellular reaction and deficiencies in collagen production. Treatment of wounds can be enhanced through systemic injection of the antibacterial drugs and/or other topical applications of medications. However, there are a number of disadvantages to these techniques, including the limited or insufficient medication penetration into the underlying skin tissue and the development of bacterial resistance with repeated antibiotic treatment. One of the more recent treatment options may involve using nanotherapeutics in combination with naturally occurring biological components, such as snail extracts (SE). In this investigation, chitosan nanoparticles (CS NPs) were loaded with an Eobania vermiculata whole-body muscle extract. The safety of the synthesized NPs was investigated in vitro to determine if these NPs might be utilized to treat full-skin induced wounds in vivo.ResultsSEM and TEM images showed uniformly distributed, spherical, smooth prepared CS NPs and snail extract-loaded chitosan nanoparticles (SE-CS NPs) with size ranges of 76–81 and 91–95 nm, respectively. The zeta potential of the synthesized SE-CS NPs was − 24.5 mV, while that of the CS NPs was 25 mV. SE-CS NPs showed a remarkable, in vitro, antioxidant, anti-inflammatory and antimicrobial activities. Successfully, SE-CS NPs (50 mg/kg) reduced the oxidative stress marker (malondialdehyde), reduced inflammation, increased the levels of the antioxidant enzymes (superoxide dismutase and glutathione), and assisted the healing of induced wounds. SE-CS NPs (50 mg/kg) can be recommended to treat induced wounds safely. SE was composed of a collection of several wound healing bioactive components [fatty acids, amino acids, minerals and vitamins) that were loaded on CS NPs.ConclusionsThe nanostructure enabled bioactive SE components to pass through cell membranes and exhibit their antioxidant and anti-inflammatory actions, accelerating the healing process of wounds. Finally, it is advised to treat rats’ wounds with SE-CS NPs.Graphical abstract

Antimicrobial activity of the interaction between essential oils against resistant Staphylococcus aureus isolates from bovine mastitis

The control of bovine mastitis is crucial for the sustainability of dairy production. Phytotherapy emerges as a potential alternative for controlling resistant bacteria and potentially replacing conventional therapies. This study aimed to assess the in vitro antimicrobial effect of the Pelargonium graveolens (GEO) and Thymus vulgaris (TEO) essential oils, as well as the interaction between them against strains of Staphylococcus aureus isolated from bovine mastitis. Eight strains of S. aureus isolated from bovine mastitis and two standard strains of human origin were used. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined using the microdilution method. The interaction between the oils was assessed using the checkerboard method. It was observed that both essential oils exhibited inhibitory effects when evaluated separately; however, TEO demonstrated superior bactericidal efficacy. The combination of both oils resulted in a bactericidal complex with a 99% reduction in the effective concentration of GEO. In conclusion, the TEO/GEO complex may contribute to improved therapeutic efficacy, reducing adverse effects, costs, and bacterial resistance development.

The scope of antimicrobial resistance in residential aged care facilities determined through analysis of Escherichia coli and the total wastewater resistome.

High and often inappropriate antibiotic use has been documented for residential aged care facilities (RACFs). As a result, RACFs represent selective environments favoring the development of antimicrobial resistance (AMR). However, surveillance of the prevalence and scope of AMR in RACFs is limited. Here, wastewater-based epidemiology encompassing culture-based methods in combination with whole genome sequencing and metagenomics allowed for the in-depth analysis of the prevalence of antimicrobial-resistant Escherichia coli in two RACFs and one retirement facility. Wastewater was collected at five different time points over 18 months. From these, E. coli were isolated and assessed for phenotypic and genotypic resistance. The antimicrobial resistome of each wastewater sample was also determined. A comparison of facilities revealed a higher prevalence of AMR and multidrug-resistant E. coli observed in one RACF. The international high-risk E. coli clone, ST131, carrying CTX-M-like extended-spectrum beta-lactamases was exclusively isolated from this facility. A high proportion of these isolates were classified as belonging to phylogroups B2 and D, associated with virulent extra-intestinal E. coli infections. The resistome of this facility also revealed a higher prevalence of mobile resistance genes such as sul1 and sul2, conferring sulfamethoxazole resistance, the plasmid-mediated quinolone-resistance gene qnrS, and qacEdelta1 that confers biocide resistance. High fluoroquinolone resistance rates were observed in all three sample sites despite measures in place limiting the use of this class of antibiotics. The findings of this study illustrate that RACFs are highly selective environments that require measures to limit AMR development, potentially through antimicrobial stewardship. IMPORTANCE Antimicrobial resistance (AMR) is a global threat that imposes a heavy burden on our health and economy. Residential aged care facilities (RACFs), where frequent inappropriate antibiotic use creates a selective environment that promotes the development of bacterial resistance, significantly contribute to this problem. We used wastewater-based epidemiology to provide a holistic whole-facility assessment and comparison of antimicrobial resistance in two RACFs and a retirement village. Resistant Escherichia coli, a common and oftentimes problematic pathogen within RACFs, was isolated from the wastewater, and the phenotypic and genotypic AMR was determined for all isolates. We observed a high prevalence of an international high-risk clone, carrying an extended-spectrum beta-lactamase in one facility. Analysis of the entire resistome also revealed a greater number of mobile resistance genes in this facility. Finally, both facilities displayed high fluoroquinolone resistance rates-a worrying trend seen globally despite measures in place aimed at limiting their use.

Quick Detection of Proteus and Pseudomonas in Patients' Urine and Assessing Their Antibiotic Susceptibility Using Infrared Spectroscopy and Machine Learning.

Bacterial resistance to antibiotics is a primary global healthcare concern as it hampers the effectiveness of commonly used antibiotics used to treat infectious diseases. The development of bacterial resistance continues to escalate over time. Rapid identification of the infecting bacterium and determination of its antibiotic susceptibility are crucial for optimal treatment and can save lives in many cases. Classical methods for determining bacterial susceptibility take at least 48 h, leading physicians to resort to empirical antibiotic treatment based on their experience. This random and excessive use of antibiotics is one of the most significant drivers of the development of multidrug-resistant (MDR) bacteria, posing a severe threat to global healthcare. To address these challenges, considerable efforts are underway to reduce the testing time of taxonomic classification of the infecting bacterium at the species level and its antibiotic susceptibility determination. Infrared spectroscopy is considered a rapid and reliable method for detecting minor molecular changes in cells. Thus, the main goal of this study was the use of infrared spectroscopy to shorten the identification and the susceptibility testing time of Proteus mirabilis and Pseudomonas aeruginosa from 48 h to approximately 40 min, directly from patients' urine samples. It was possible to identify the Proteus mirabilis and Pseudomonas aeruginosa species with 99% accuracy and, simultaneously, to determine their susceptibility to different antibiotics with an accuracy exceeding 80%.

Newly isolated phages preying on Pseudomonas syringae pv. garcae: In vitro and ex vivo inactivation studies in coffee plant leafs

Coffee canker, or bacterial halo blight (BHB) of coffee, is a disease caused by the phytopathogenic bacterium Pseudomonas syringae pv. garcae (Psg), having been found for the first time in 1955, in the Garça region (State of São Paulo), and which has stood out in the Brazilian coffee plantations in recent years, leading to severe economic losses that seriously affect coffee trade. The treatments available are still scarce, involving frequent spraying of coffee plantations with either copper derivatives or the antibiotic kasugamycin. However, these compounds should be avoided due to environmental toxicity and the development of bacterial resistances. Herein we report the isolation and physical/biological characterisation of two novel lytic phages and their efficacy in the control of Psg. Phages ph002F and ph004F were isolated from coffee plant leaves in Brazil (Sorocaba/SP and Itu/SP cities), using Psg IBSBF-158 as the host. According to the transmission electron microscopy analyses, both phages belong to the class Caudoviricetes and present myovirus-like morphotypes. Phages ph002F and ph004F showed eclipse times of 5 min and 20 min, respectively, and a burst size of 123 PFU/host cell and 12 PFU/host cell, respectively, allowing to conclude they replicate well in Psg IBSBF-158 with latency periods of 50 min. Phage ph002F (reduction of 4.59 log CFU/mL, compared to uninfected culture) was more effective in inactivating Psg than phage ph004F (reduction of 3.85 log CFU/mL) after 10 h of incubation at a MOI of 10. As a cocktail, the two phages were highly effective in reducing the bacterial load (reduction of 5.26 log CFU/mL at a MOI of 0.1 or reduction of 5.03 log CFU/mL at a MOI of 10, relative to untreated culture), after 12 h of treatment. This study provides evidence that the isolated phages are promising candidates against the causative agent of BHB in coffee plants.

Dual Functional Diblock Amino Acid Copolymer Displaying Synergistic Effect with Curcumin against MRSA and Encapsulation of Curcumin†

Comprehensive SummaryThe intensive use of antibiotics intensifies the development of bacterial resistance, which has become a serious problem globally. Methicillin‐resistant Staphylococcus aureus (MRSA) has resulted in significant morbidity and mortality. Therefore, it is an urgent need to develop new antimicrobial drugs and administration methods. Herein, we report a dual functional diblock copolymer PLL20‐b‐PBLG20, which was prepared by superfast and water‐insensitive polymerization on N‐carboxyanhydrides (NCA) initiated by tetraalkylammonium carboxylate. In addition to direct antimicrobial activity, PLL20‐b‐PBLG20 also exerts a synergistic bactericidal effect against MRSA with curcumin, a plant extract with antibacterial property. Moreover, PLL20‐b‐PBLG20 successfully encapsulates curcumin to form nanoparticles via self‐assembly. The combination of dual functional PLL20‐b‐PBLG20 and curcumin holds promise in combating MRSA infections.

Nutrient condition modulates the antibiotic tolerance of Pseudomonas aeruginosa

The variation in nutrient content across diverse environments has a significant impact on the survival and metabolism of microorganisms. In this study, we examined the influence of nutrients on the antibiotic tolerance of the PAO1 strain of Pseudomonas aeruginosa. Our findings indicate that under nutrient-rich conditions, this strain exhibited relatively high tolerance to ceftazidime, chloramphenicol, and tetracycline, but not aminoglycosides and fluoroquinolones. Transcriptome analysis revealed that genes associated with antibiotic tolerance were expressed more efficiently in nutrient-rich media, including ribosomal protein genes and multidrug efflux pump genes, which conferred higher tetracycline tolerance to the strain. Furthermore, the genes responsible for translation, biosynthesis, and oxidative phosphorylation were suppressed when nutrients were limited, resulting in decreased metabolic activity and lower sensitivity to ciprofloxacin. Artificial interference with ATP synthesis utilizing arsenate confirmed that the curtailment of energy provision bolstered the observed tolerance to ciprofloxacin. In general, our results indicate that this strain of P. aeruginosa tends to activate its intrinsic resistance mechanisms in nutrient-rich environments, thereby enhancing resistance to certain antibiotics. Conversely, in nutrient-limited environments, the strain is more likely to enter a dormant state, which enables it to tolerate antibiotics to which it would otherwise be sensitive. These findings further suggest that antibiotics released in environments with varying eutrophication levels may have divergent effects on the development of bacterial antibiotic resistance.

Synthesis, in Silico Study and Biological Evaluation of N-(Benzothiazol/Thiazol-2-yl)benzamide Derivatives as Quorum Sensing Inhibitors against Pseudomonas aeruginosa.

The development of bacterial resistance to chemical therapy poses a severe danger to efficacy of treating bacterial infections. One of the key factors for resistance to antimicrobial medications is growth of bacteria in biofilm. Quorum sensing (QS) inhibition was created as an alternative treatment by developing novel anti-biofilm medicines. Cell-cell communication is impeded by QS inhibition, which targets QS signaling pathway. The goal of this work is to develop newer drugs that are effective against Pseudomonas aeruginosa by decreasing QS and acting as anti-biofilm agents. In this investigation, N-(benzo[d]thiazol-2-yl)benzamide/N-(thiazol-2-yl)benzamide derivatives 3a-h were designed and synthesized in good yields. Further, molecular docking analyses revealed that binding affinity values were founded -11.2 to -7.6 kcal/mol that were moderate to good. The physicochemical properties of these prepared compounds were investigated through in-silico method. Molecular dynamic simulation was also used to know better understanding of stability of the protein and ligand complex. Comparing N-(benzo[d]thiazol-2-yl)benzamide 3a to salicylic acid (4.40±0.10) that was utilised as standard for quorum sensing inhibitor, the anti-QS action was found greater for N-(benzo[d]thiazol-2-yl)benzamide 3a (4.67±0.45) than salicylic acid (4.40±0.10). Overall, research results suggested that N-(benzo[d]thiazol-2-yl)benzamide/N-(thiazol-2-yl)benzamide derivatives 3a-h may hold to develop new quorum sensing inhibitors.

Membrane-Active Metallopolymers: Repurposing and Rehabilitating Antibiotics to Gram-Negative Superbugs.

Among multiple approaches to combating antimicrobial resistance, a combination therapy of existing antibiotics with bacterial membrane-perturbing agents is promising. A viable platform of metallopolymers as adjuvants in combination with traditional antibiotics is reported in this work to combat both planktonic and stationary cells of Gram-negative superbugs and their biofilms. Antibacterial efficacy, toxicity, antibiofilm activity, bacterial resistance propensity, and mechanisms of action of metallopolymer-antibiotic combinations are investigated. These metallopolymers exhibit4-16-fold potentiation of antibiotics against Gram-negative bacteria with negligible toxicity toward mammalian cells. More importantly, the lead combinations (polymer-ceftazidime and polymer-rifampicin) eradicate preformed biofilms of MDR E. coli and P. aeruginosa, respectively. Further, β-lactamase inhibition, outer membrane permeabilization, and membrane depolarization demonstratesynergy of these adjuvants with different antibiotics. Moreover, the membrane-active metallopolymers enablethe antibiotics to circumvent bacterial resistance development. Altogether, the results indicatethat such non-antibiotic adjuvants bear the promise to revitalize the efficacy of existing antibiotics to tackle Gram-negative bacterial infections.