Biofilm Inhibitory Effect Research Articles

A bifunctional nanocomplex with remineralizing and antibacterial activities to interrupt dental caries

Dental caries result from an imbalance between the demineralization and remineralization of dental hard tissues, primarily caused by biofilm accumulation. According to the theory of interrupting dental caries, effective anticaries materials and techniques should possess both remineralizing and antibacterial properties. However, current anticaries materials cannot replicate amelogenesis, which is crucial for remineralization and for preventing bacterial adhesion and biofilm formation. In this study, silk fibroin (SF) loaded with benzalkonium chloride (BZC) successfully formed an SF-BZC composite. This composite stabilized amorphous calcium phosphate (ACP), creating an ACP@SF-BZC dual-functional nanocomplex with both remineralizing and antibacterial properties. ACP@SF-BZC demonstrated significant anti-adhesion and biofilm inhibitory effects against Streptococcus mutans and Streptococcus sobrinus. Moreover, compared to fluoride, ACP@SF-BZC significantly enhanced the remineralization of demineralized enamel surfaces, forming a stable remineralized layer with improved mechanical properties, both in vitro and in vivo. In summary, the dual-function ACP@SF-BZC nanocomplex, with its remineralizing and antibacterial effects, offers a promising alternative for preventing and arresting enamel caries.

Antimicrobial, Antibiofilm and Anticancer Potentials of Glycine and Glycyl-Glycine; an in vitro study

Globally, there is a huge demand for novel agents capable of providing protection against both pathogen microorganisms and tumor cells. In this study, the antimicrobial, biofilm inhibitory, and anticancer effects of glycine and glycyl-glycine were investigated. The antimicrobial effects were determined using the broth dilution method, while the biofilm inhibitory effects were assessed through the crystal violet binding assay. Cytotoxic effects on HeLa cell viability were measured using the MTT assay. Our results indicate that, although 100 mg/mL of glycine only inhibited S. epidermidis W17 among the three tested isolates, 400 mg/mL of glycyl-glycine inhibited both S. epidermidis W17 and P. mirabilis U15 strains. Additionally, sub-MICs (concentrations below the Minimum Inhibition Concentration) of glycine inhibited biofilm formation by more than 70% in all tested clinical isolates and exhibited significantly more biofilm inhibition against P. mirabilis U15 and S. epidermidis W17 strains (p

Synergy of Histone Acetyltransferase Inhibitor (HATi) with Quercetin Inhibits Biofilm Formation in Candida tropicalis.

Histone acetyltransferase inhibitors (HATi) are mechanism-based inhibitors that show promise in the treatment of several illnesses, including diabetes, hyperlipidemia, cancer, and Alzheimer's disease. The work emphasizes the significance of HATi as a possible treatment strategy against Candida species biofilms. Here, in this study, we found that combining a HATi, anacardic acid, and quercetin, a known flavonoid, significantly prevented biofilm formation by C. tropicalis. We further show that C. tropicalis exhibited a considerable downregulation of drug-resistance gene expression (CDR1 and MDR1) when co-administrated. Additionally, in silico studies revealed that the anacardic acid (AA) interacts strongly with a histone acetyltransferase, Rtt109, which may account for the observed biofilm inhibitory effect. In conclusion, the study illustrates how HATi may be used to potentiate the inhibitory action of phytoactives or antifungals against drug-resistant yeast infections.

The prebiotic effects of fructooligosaccharides enhance thegrowth characteristics of Staphylococcus epidermidis and enhance the inhibition of Staphylococcus aureus biofilm formation.

Oligosaccharides have been shown to enhance the production of short chain fatty acids (SCFAs) by gut probiotics and regulate gut microbiota, to improve intestinal health. Recent research indicates that oligosaccharides may also positively impact skin microbiota by selectively promoting the growth of skin commensal bacteria and inhibiting pathogenic bacteria. However, the specific metabolic and regulatory mechanisms of skin commensal bacteria in response to oligosaccharides remain unclear. This study aims to explore the influence of four oligosaccharides on the growth and metabolism of Staphylococcus epidermidis and further identify skin prebiotics that can enhance its probiotic effects on the skin. Fructooligosaccharides (FOS), isomaltooligosaccharide (IMO), galactooligosaccharides (GOS) and inulin were compared in terms of their impact on cell proliferation, SCFAs production of S. epidermidis CCSM0287 and the biofilm inhibition effect of their fermentation supernatants on Staphylococcus aureus CCSM0424. Furthermore, the effect of FOS on S. epidermidis CCSM0287 was analysed by the transcriptome analysis. All four oligosaccharides effectively promoted the growth of S. epidermidis CCSM0287 cells, increased the production of SCFAs, with FOS demonstrating the most significant effect. Analysis of the SCFAs indicated that S. epidermidis CCSM0287 predominantly employs oligosaccharides to produce acetic acid and isovaleric acid, differing from the SCFAs produced by gut microbiota. Among the four oligosaccharides, the addition of 2% FOS fermentation supernatant significantly inhibited S. aureus CCSM0424 biofilm formation. Furthermore, RNA sequencing revealed 162 differentially expressed genes (84 upregulated and 78 downregulated) of S. epidermidis CCSM0287 upon FOS treatment compared with glucose treatment. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis highlighted differences in the amino acid synthesis pathway, particularly in terms of arginine biosynthesis. FOS promotes cell proliferation, increases the SCFA production of S. epidermidis CCSM0287 and enhance the inhibition of S. aureus biofilm formation, suggesting that FOS serves as a potential prebiotic for strain S. epidermidis CCSM0287.

Self-adhesive poly-l-lysine/tannic acid hybrid hydrogel for synergistic antibacterial activity against biofilms

Biomedical implants are crucial for enhancing various human physiological functions. However, they are susceptible to microbial contamination after implantation, posing a risk of implant failure. To address this issue, hydrogel-based coatings are used, but achieving both effective antibacterial properties and stable adhesion remains challenging. This study introduces a hybrid hydrogel network made from Tannic Acid (TA) and Poly-l-Lysine (PLL), cross-linked through ionic and hydrogen bonds, which imparts adhesive and anti-infective properties. The physicochemical analysis revealed that the hydrogels exhibited significant porosity, favorable mechanical characteristics, and demonstrated in vitro enzymatic biodegradation. Moreover, the hydrogels demonstrated adhesion to various substrates, including Ti alloy with an adhesive strength of 42.5 kPa, and retained their integrity even after immersion in water for a minimum of 10 days. The modified Ti surfaces significantly reduced protein adsorption (∼70 %), indicating antifouling properties. The hydrogels prevented bacterial adhesion on titanium surfaces through a “contact-kill” mode of action and inhibited biofilm formation by around 94.5 % for Staphylococcus aureus and 90.8 % for Pseudomonas aeruginosa. The modified Ti retained biofilm inhibitory effects for at least six days without significant performance decline. In vitro cytotoxicity assay confirmed the biocompatibility of the hydrogels with NIH3T3 cells. Overall, these results highlight the competence of hybrid hydrogels as effective coatings for Ti implants, offering strong adhesion and biofilm prevention to mitigate implant-related infections.

Development of poly(lactic acid)-based natural antimicrobial film incorporated with caprylic acid against Salmonella biofilm contamination in the meat industry

Using a solvent-casting method, a poly(lactic acid) (PLA) film incorporated with caprylic acid (CA) was developed as an active packaging against Salmonella enterica ser. Typhimurium and S. enteritidis to reduce the risk of microbial contamination during distribution and storage of meat. According to the minimum inhibitory concentration (MIC) test results of the natural antimicrobial, CA was introduced at 0.6, 1.2, 2.4, and 4.8 % (v/v) into neat PLA. The biofilm inhibitory effect and antimicrobial efficacy of CA-PLA film against both Salmonella strains, as well as the intermolecular interactions and barrier properties of CA-PLA film, were evaluated. Biofilm formation was reduced to below the detection limit (<1.0 log CFU/cm2) for both S. typhimurium and S. enteritidis when co-cultured overnight with 4.8 % CA-PLA film. The 4.8 % CA-PLA film achieved maximum log reductions of 2.58 and 1.65 CFU/g for S. typhimurium and 2.59 and 1.76 CFU/g for S. enteritidis on inoculated chicken breast and beef stored at 25 °C overnight, respectively, without any quality (color and texture) losses. CA maintained its typical chemical structure in the film, as confirmed by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectra. Furthermore, film surface morphology observations by field emission scanning electron microscopy (FESEM) showed that CA-PLA film was smoother than neat PLA film. No significant (P > 0.05) changes were observed for water vapor permeability and oxygen permeability by the addition of CA into PLA film, suggesting that CA-PLA film is a promising strategy for active packaging to control Salmonella contamination in the meat industry.

Tannic acid inhibits Escherichia coli biofilm formation and underlying molecular mechanisms: Biofilm regulator CsgD

Biofilms often engender persistent infections, heightened antibiotic resistance, and the recurrence of infections. Therefor, infections related to bacterial biofilms are often chronic and pose challenges in terms of treatment. The main transcription regulatory factor, CsgD, activates csgABC-encoded curli to participate in the composition of extracellular matrix, which is an important skeleton for biofilm development in enterobacteriaceae. In our previous study, a wide range of natural bioactive compounds that exhibit strong affinity to CsgD were screened and identified via molecular docking. Tannic acid (TA) was subsequently chosen, based on its potent biofilm inhibition effect as observed in crystal violet staining. Therefore, the aim of this study was to investigate the specific effects of TA on the biofilm formation of clinically isolated Escherichia coli (E. coli). Results demonstrated a significant inhibition of E. coli Ec032 biofilm formation by TA, while not substantially affecting the biofilm of the ΔcsgD strain. Moreover, deletion of the csgD gene led to a reduction in Ec032 biofilm formation, alongside diminished bacterial motility and curli synthesis inhibition. Transcriptomic analysis and RT-qPCR revealed that TA repressed genes associated with the csg operon and other biofilm-related genes. In conclusion, our results suggest that CsgD is one of the key targets for TA to inhibit E. coli biofilm formation. This work preliminarily elucidates the molecular mechanisms of TA inhibiting E. coli biofilm formation, which could provide a lead structure for the development of future antibiofilm drugs.

Chemical characterization of Thymus zygis L. (thyme) and Elettaria cardamomum L. (cardamom) essential oils and investigation of their inhibitory potentials against cervicovaginal Escherichia coli isolates

IntroductionThe cervicovaginal microbiota plays a crucial role in women's well-being and reproductive health. This study initially focuses on identifying the chemical components of the essential oils (EOs) from Elettaria cardamomum L. (cardamom) (Ec) and Thymus zygis L. (thyme) (Tz). Subsequently, this study aims to determine the antimicrobial and biofilm inhibitory effects of these EOs against both cervicovaginal mixed cultures and the microbial strains isolated from them. MethodsThe compositions of EOs were determined by GC–MS analysis using an Agilent 7890B GC/5977A Series MSD System. Cervicovaginal samples were obtained from two women presenting complaints of a strong odor and vaginal discharge. Samples were collected from the cervix using a sterile cytobrush, ensuring no contact with the urethra, rectum, or external parts of the vagina. The cytological examination was conducted using Papanicolaou staining. Isolated microorganisms were identified through 16S Ribosomal RNA Gene Sequence Analysis and characterized using antibiogram tests. Antimicrobial tests of the EOs were performed by broth microdilution and disc diffusion methods. Biofilm formation was assessed using a crystal violet binding assay. ResultsThe major components of Ec EO were α-terpinyl acetate (41.95 %) and eucalyptol (28.58 %), whereas Tz EO contained significant amounts of o-cymene (38.76 %), carvacrol (22.66 %), and thymol (20.69 %). Escherichia coli codes OR651248 and OR651249 were isolated from CV 6 and CV 7 mixed cultures, respectively. E. coli OR651248 demonstrated a higher antibiotic resistance level than E. coli OR651249. Biofilm formation by mixed cultures was significantly higher than that by E. coli isolates (P < 0.05). The Ec EO displayed minimal to no effect against both mixed cultures, showing zero inhibition zone, 500 μL/mL minimum inhibitory concentrations (MICs), and 500 μL/mL minimum microbicidal concentrations (MMCs). However, the Tz EO exhibited antimicrobial activity against both mixed cultures and the E. coli isolates, with inhibition zones > 15 mm and MICs/MMCs < 4 μL/mL (P < 0.05). In all microbial cultures, 0.5 MIC of both EOs inhibited biofilm formation by more than 80 %. ConclusionThe Tz EO exhibits promising potential as an antimicrobial and biofilm inhibitory agent for managing cervicovaginal E. coli infections.

Exploring anti-quorum sensing potential and biofilm inhibitory effect of azithromycin against urinary Escherichia coli isolated from a teaching hospital in Alexandria

Exploring anti-quorum sensing potential and biofilm inhibitory effect of azithromycin against urinary Escherichia coli isolated from a teaching hospital in Alexandria

The Potential Teeth Bleaching and Halitosis Prevention Effects of Pediococcus inopinatus THK-30, a Kimchi-Derived Lactic Acid Bacterium: In Vitro Study.

Recent developments in addressing dental aesthetic concerns, encompassing issues like teeth discoloration and halitosis, underscore the demand for safer alternative solutions. This study aims to confirm the effects of lactic acid bacteria (LAB) from kimchi on artificial teeth bleaching and their potential impact in terms of preventing halitosis-related bacteria. To evaluate the antimicrobial effects against oral pathogens, disc diffusion tests and broth microdilution methods were used. Additionally, crystal violet analysis was performed to confirm the biofilm inhibition effect. The bleaching effects on stained artificial teeth were analyzed using the CIEDE2000 colorimetric method. Statistical analyses were performed using GraphPad Prism 9 with one-way and two-way ANOVA, with the significance level set at α < 0.05. The strain THK-30, isolated from kimchi, exhibited antibacterial activity against Streptococcus mutans, Porphyromonas gingivalis, and Fusobacterium nucleatum, and was identified as Pediococcus inopinatus. Moreover, THK-30 showed a synergistic antibacterial effect against Gram-negative oral pathogens with 8% sodium hexametaphosphate (SHMP). In the stained artificial teeth bleaching test and artificial teeth biofilm inhibition test, the cell-free supernatant of THK-30 displayed significant teeth bleaching effects and caused the inhibition of biofilm formation, both independently and in combination with SHMP 8%. This study has demonstrated the potential applicability of LAB in teeth discoloration and halitosis. These findings are poised to provide a foundation for the development of research pertaining to the control of oral bacteria.

Investigation of advanced biological properties of carbon, carbon‐boron quantum dots, and copper (II) phthalocyanine nanoconjugates

Non‐peripherally, glycerol terminal groups substituted copper (II) phthalocyanine were non‐covalently (electrostatic and/or π–π interaction) attached to carbon (CQD) and carbon‐boron quantum dots (CBQD) to form QDs‐Pc nanoconjugate systems. Synthesized novel phthalocyanine compounds and QDs‐Pc conjugate systems were characterized using different spectroscopic techniques. Various biological assessments were applied to newly synthesized compounds. Conjugates 4 and 5 had a maximal free radical scavenging activity of 71.3% and 68.1% at a 100 mg/L concentration. Compounds exhibited high antidiabetic activities at 200 mg/L. Also, compounds showed significant DNA nuclease activity at all tested concentrations. The most efficient MIC value was obtained as 4 mg/L against Enterococcus hirae and Enterococcus feacalis. This MIC value was further decreased after photodynamic therapy, and it was observed that the antimicrobial effects of the compounds increased. Inhibition of microbial cell viability was obtained as 100% for all compounds. In addition, compounds exerted perfect biofilm inhibitory effects.

تأثير دقائق اوكسيد الزنك النانوية المصنعة بايولوجيًا على النمط الظاهري والجيني للغشاء الحيوي لبكتريا Proteus mirabilis

تصنف بكتريا Proteus mirabilis كثالث مسبب لالتهاب المجاري البولية المصاحب لتركيب أنابيب القسطرة البولية. تعد الاغشية الحيوية من عوامل الضراوة اللتي تستعملها البكتريا لمقاومة المضادات الحيوية وفي ضل تطور مقاومة البكتريا للمضادات الحيوية, صار لزامًا على العلماء والباحيثن في مجال الاحياء المجهرية أيجاد بدائل ذات كفائة عالية قادرة على اختراق جدار الخلية البكتيرية وبالتالي قتلها.في السنوات الاخيرة, اكتسبت دقائق النانو المصنعة بايولوجيا والصديقة للبيئة أهتمامًا كبيرًا من قبل الباحثين لأمتلاكها خصائص مميزة كصغر حجمها المتناهي واشكالها المختلفة يمَكنهم من استعمالها كمواد مضادة للبكتيريا في مختلف المجلات الطبية والصحية. تهدف هذه الدراسة الى التحري عن قدرة دقائق اوكسيد النحاس النانوية المصنعة بايولوجيا على تثبيط تكوين الغشاء الحيوي المنتج من قبل بكتيريا Proteus mirabilis. أنتجت هذه الدقائق بأستعمال راشح بكتيري يحتوي على الانزيمات المختزلة لملح النحاس واستخدمت عدة تقنيات لتحديد الصفات الفيزيائية المظهرية لدقائق النانو المنتجة حيث استخدمت تقنية XRD لتأكيد التركيب خماسي الشكل لهذه الدقائق , بينما استعملت تقنية TEM لتحديد قطر الجزيئات المنتجة بمعدل 84.45 نانومتر. اما FESEM فقد تم استخدامه لتحديد الصفات الخارجية لسطح جزيئات النانو المنتجة , تم تأكيد النتائج بأستخدام تقنية AFM للكشف عن توزيع و وعورة جزيئات النانو.اما UV فقد سجلت ااقصى امتصاصية على الطول الموجي nm287 .تم التحري في هذه الدراسة عن فعالية دقائق اوكسيد النحاس النانوية في منع تكون الغشاء الحيوي حيث أستعمل مستحلب دقائق اوكسيد النحاس النانوي بالتركيز تحت القاتل 32 μg/ml . سجلت نتائج هذه الدراسة انخفاضا ملحوضَا في انتاج الغشاء الحيوي للبكتريا المنتجه له حيث سجلت العزلات ذات الانتاجية العالية للغشاء الحيوي ضعفأ في الانتاج بعد معاملتها بالتركيز المستعمل من دقائق اوكسيد النحاس النانوية. جاء تأثير هذه الدقائق على كل من النمط الظاهري والتعبير الجيني لجين LuxS المسؤول عن تكوين الغشاء الحيوي بأستخدام تقنية rtPCRحيث سجلت الدراسة انخفاضا في التعبير الجيني لهذا الجين بعد معاملة سلالات البكتريا قيد الدراسة بدقائق اوكسيد النحاس النانوية مقارنتاً بالعزلات غير المعاملة. من خلال نتائج هذه التجربة يمكن التوصل الى امكانية اعتماد دقائق اوكسيد الزنك النانوية كعامل مضاد للبكتيريا عن استخدامه بالتركيز المناسب.

Antifouling activity of aquatic macrophyte extracts on estuarine bacterial biofilms

ABSTRACT Ecologically safe antifouling solutions have received growing attention since the acknowledgment of the environmental risks of traditional biocides used currently. Given that bacteria constitute the initial stage of the biofouling process, this work aims to evaluate the antifouling potential of aquatic macrophyte extracts in inhibiting biofilm-forming bacteria and to evaluate their ecotoxicological effects on target and nontarget organisms (Thalassiosira pseudonana and Nitokra sp.). For this, the ability of 25 macrophyte extracts to inhibit and eradicate single and multispecies estuarine biofilms was evaluated. The extracts and respective serial dilutions that showed a biofilm inhibitory effect ≥ 60% were evaluated in ecotoxicological assays. Of the 25 extracts, only 6 extracts inhibited the biofilm ≥ 60%. The extracts of Cabomba caroliniana and Schoenoplectus californicus stood out as having a biofilm inhibitory effect ≥ 80% (p < 0.05) for bacterial isolates and ≥ 70% (p < 0.05) for multispecies biofilms. Furthermore, these extracts showed no toxic effects on nontarget organisms. These results demonstrate the biotechnological potential of macrophyte compounds.

LACTIC ACID BACTERIA AND FERMENTED MAIZE SUPERNATANT (Omidun) HAVE ANTI-BIOFILM PROPERTIES AGAINST STAPHYLOCOCCI AND ENTEROAGGREGATIVE Escherichia coli STRAINS

Bacterial infections caused by biofilm forming organisms are of public health concern due to their propensity to contribute to persistent chronic diseases, chiefly because of their ability to resist antibiotics and host immune functions. Probiotics are considered useful therapeutic option in combating pathogenic biofilms. This study evaluates the anti-biofilm properties of potential probiotic Lactic Acid Bacteria (LAB) and fermented maize supernatant (Omidun) against selected biofilm-forming pathogens. Crystal violet biofilm assay was used to determine LAB and Omidun biofilm inhibition and dispersion in selected pathogens (Pseudomonas aeruginosa, Coagulase-negative staphylococci (CoNS), S. aureu and Enteroaggregative Escherichia coli) at different concentration (1%, 10%, 50%, 100%) of neutralized and non-neutralized cell free supernatant (CFS). Percentage biofilm inhibitions and dispersions were evaluated, and data were analysed with ANOVA. Omidun and LAB showed promising biofilm inhibitory and dispersive effect against the selected pathogens. L. plantarum showed the greatest biofilm inhibitory effect (P. aeruginosa: 7.85%, CoNS: 27.75%, S. aureus: 66.90%, EAEC: 39.73%) and dispersive effect (P. aeruginosa: 15.94%, CoNS: 23.27%, S. aureus: 24.90%, EAEC: 32.09%) against the selected pathogens while Omidun showed the least biofilm inhibitory and dispersive effect against the selected pathogens. There was no significance difference in the percentage of biofilm inhibition and dispersion produced under different concentrations, neutralized and non-neutralized state. Pseudomonas aeruginosa was the most resistant pathogen while Enteroaggregative Escherichia coli (EAEC) was the most susceptible. Inhibition and dispersion of biofilm can be mediated by LAB and Omidun, these effects appear to be independent of the produced organic acids

Inhibitory Effects of Biosynthesized Copper Nanoparticles on Biofilm Formation of Proteus mirabilis

This study aimed to explore whether green synthesized copper nanoparticles (CuNPs) can function as an anti-biofilm agent produced by P. mirabilis. The nanoparticles were synthesized from cells free extract of P. mirabilis. Characterization of biosynthesized copper nanoparticles was carried out to determine the chemical and physical properties of the product using atomic-force microscopy (AFM), transmission electron microscopy (TEM), field emission scanning electron microscopy (FESEM), X-ray diffraction (XRD) and UV-visible spectroscopy. The hexagonal structure was confirmed by XRD, size range was marked 13-19nm by TEM. FESEM was used to confirm the surface morphology. AFM analysis was used to reveal the roughness and distribution of nanoparticles. UV–visible spectra of the synthesized nanoparticles recorded a maximum peak at 380 nm. Copper nanoparticles showed remarkable biofilm inhibitory effects on wild type strains of multidrug resistant Proteus mirabilis. After incubation for 24 and 48 hours at 37oC with 128 μg/ml sub minimum inhibitory concentration (MIC) of CuO nanoparticles, strong biofilm producer strains showed weak biofilm production. Down regulation changes in LuxS expression using real time PCR technology were detected after treatment with copper nanoparticles of these strains.

Drug repurposing: insights into the antimicrobial effects of AKBA against MRSA

Staphylococcus aureus is a major threat in infectious diseases due to its varied infection types and increased resistance. S. aureus could form persister cells under certain condition and could also attach on medical apparatus to form biofilms, which exhibited extremely high resistance to antibiotics. 3-Acetyl-11-keto-beta-boswellic acid (AKBA) is a well-studied anti-tumor and antioxidant drug. This study is aimed to determine the antimicrobial effects of AKBA against S. aureus and its persister cells and biofilms. The in vitro antimicrobial susceptibility of AKBA was assessed by micro-dilution assay, disc diffusion assay and time-killing assay. Drug combination between AKBA and conventional antibiotics was detected by checkerboard assay. And the antibiofilm effects of AKBA against S. aureus were explored by crystal violet staining combined with SYTO/PI probes staining. Next, RBC lysis activity and CCK-8 kit were used to determine the cytotoxicity of AKBA. In addition, murine subcutaneous abscess model was used to assess the antimicrobial effects of AKBA in vivo. Our results revealed that AKBA was found to show effective antimicrobial activity against methicillin-resistant S. aureus (MRSA) with the minimal inhibitory concentration of 4–8 µg/mL with undetectable cytotoxicity. And no resistant mutation was induced by AKBA after 20 days of consecutive passage. Further, we found that AKBA could be synergy with gentamycin or amikacin against S. aureus and its clinical isolates. By crystal violet and SYTO9/PI staining, AKBA exhibited strong biofilm inhibitory and eradication effects at the concentration of 1 ~ 4 µg/mL. In addition, the effective antimicrobial effect was verified in vivo in a mouse model. And no detectable in vivo toxicity was found. These results indicated that AKBA has great potential to development as an alternative treatment for the refractory S. aureus infections.

Antifungal Biofilm Inhibitory Effects of Combinations of Diclofenac and Essential Oils.

Systemic fungal infections have risen in recent decades and most of them are caused by Candida species, which are becoming increasingly resistant to conventional antifungal drugs. Biofilm production has been considered the most common growth form of Candida cells and is associated with a high level of antifungal resistance. At present, international research reports on the antifungal activity of non-traditional antimicrobial drugs and their potential use against life-threatening resistant fungal infections. Indeed, drug repurposing has led to the consideration of well-known compounds as a last-line therapy. The goal of this work is to evaluate the potential synergistic antifungal biofilm activity of new combinations between diclofenac sodium salt (DSS), a widely used non-steroidal anti-inflammatory drug (NSAID), with the essential oils (EOs) of Mentha piperita, Pelargonium graveolens, and Melaleuca alternifolia, whose antifungal activity has been well documented over the years. The in vitro antifungal activity of DSS and EOs was determined on different Candida strains. Susceptibility testing and the synergism of DSS and EOs versus biofilm cells was performed by using the broth microdilution assay and checkerboard methods. Minimum inhibitory concentrations (sMIC50) of DSS alone ranged from 1.25 to 2.05 mg/mL for all the strains considered. These values significantly decreased when the drug was used in combination with the EOs. The fractional inhibitory concentration index (FICI) was lower than 0.5 for almost all the associations, thus indicating a significant synergism, particularly for the DSS-Pelargonium graveolens combination towards the Candida strains examined. These preliminary results show that the combination of the EOs with DSS improves the antifungal activity on all the tested Candida strains, significantly lowering the concentrations of the components used and thus allowing any toxic effects to be overcome.

Biofilm Inhibitory Effects of Lactobacillus Spp Against Streptomycin-resistant Uropathogenic Escherichia Coli

The biofilm inhibitory effects of Lactobacillus spp against Streptomycin-resistant uropathogenic Escherichia coli (UPEC) were evaluated using the crystal violet test method. Lactobacillus spp were isolated from milk samples while fifty strains of Uropathogenic Escherichia coli were isolated from urine samples from Urinary Tract Infection patients attending Federal Medical Centre (FMC) Owerri, Nigeria. Ten of the E. coli strains resistant to streptomycin antibiotics were screened for their susceptibility to antibiofilm effect of Lactobacillus secondary metabolites extracts. From the result obtained, only one of the E. coli strains was susceptible while nine strains were resistant. This result shows clearly that the metabolite extracts from Lactobacillus sp were not effective in the antibiofilm activity of the E. coli strains and thus not a good candidate for the management of UTI caused by E. coli.

Antimicrobials discovery against Staphylococcus aureus by high throughput screening of drug library

To develop antimicrobials against Staphylococcus aureus by high throughput screening of drug library. The type of this study is experimental research. The clinical isolates of S. aureus were collected from the sputum samples of respiratory inpatient department of the Third Xiangya Hospital of Central South University. The anti-planktonic cells growth inhibition activity of FDA-approved drugs library (including 1 573 molecules) was assessed by building a planktonic cells screening platform; The biofilm inhibitory effect of the FDA-approved drugs was detected by building a biofilm screening platform combined with crystal violet staining; Minimal inhibitory concentrations of the selected hits were determined by broth microdilution assay. Finally, the cytotoxicity of the selected hits was detected by CCK-8 assay. The results showed that 218 hits were exhibited effective growth inhibitory effects against S. aureus by setting the concentrations of the molecules in the FDA-approved library to 100 μmol/L. These selected molecules are mainly anti-infective drugs, accounting for 118 hits; Followed by anti-cancer drugs, anti-inflammatory/-immune drugs, neurological drugs, cardiovascular drugs, endocrine drugs, and metabolic disease drugs, which accounts for 40, 19, 12, 9, 8, and 3 hits; Other unclassified drugs accounts for 9 hits. The top 10 hits exhibiting anti-planktonic cells activity against S. aureus were mainly including antitumor drugs, followed by neurological drugs and unclassified drugs like vitamin K3 with the inhibition rate of 99.65%-100%. Similarly, the top 10 hits showing biofilm inhibitory effects against S. aureus were also mainly including antitumor drugs, followed by neurological drugs and anti-inflammatory/-immune drugs with the inhibition rate of 50.22%-92.95%. The minimal inhibitory concentration (MIC) of the 51 hits by second round screening was determined by micro-dilution assay, which mainly include the antitumor drugs, cardiovascular drugs, endocrine drugs, anti-inflammatory/-immune drugs, metabolic disease drugs, neurological drugs and other unclassified drugs accounted for 22, 5, 3, 9, 2, 5 and 5 hits, respectively, with the MICs of 1.56-50 μmol/L, 6.25-25 μmol/L, 6.25-25 μmol/L, 0.2-50 μmol/L, 25-50 μmol/L, 1.56-50 μmol/L and 0.1-12.5 μmol/L, respectively. In conclusion, the minimum inhibitory concentrations of small molecules screened through high-throughput assay are at the level of micromolar with strong drug development potential and high modifiability. The high effective anti-planktonic cells and anti-biofilm activity by these molecules are expected to provide new ideas for the development of new antimicrobials against S. aureus.

Benzoic acid derivatives as potent antibiofilm agents against Klebsiella pneumoniae biofilm

Klebsiella pneumoniae is responsible for a significant proportion of human urinary tract infections, and its biofilm is a major virulence. One potential approach to controlling biofilm-associated infections is targeting the adhesin MrkD1P to disrupt biofilm formation. We employed Schrodinger's Maestro tool with the OPLS 2005 force field to dock compounds with the target protein. Two benzoic acid derivatives, 3-hydroxy benzoic acid and 2,5-dihydroxybenzoic acid, had strong binding free energies (-55.57 and-18.68kcal/mol) and were the most potent compounds. The in-vitro experiments were conducted to validate the in-silico results. The results showed that both compounds effectively inhibited biofilm formation at low concentrations (4 and 8mg/mL, respectively) and had antibiofilm activity, restricting cell attachment. Both compounds demonstrated a strong biofilm inhibitory effect, with 97% and 89% reduction in biofilm by 3-hydroxy benzoic acid and 2,5-dihydroxybenzoic acid, respectively. These findings suggest that natural compounds can be a potential source of new drugs to combat biofilm-associated infections. The study highlights the potential of targeting adhesin MrkD1P as an effective approach to controlling biofilm-associated infections caused by K.pneumoniae. The results may have implications for the development of new therapies for biofilm-associated infections and pave the way for future research in this area.