Search Results (322)

Candida infections pose a significant health threat, and conventional antifungal drugs like nystatin are limited due to poor solubility, skin permeability, and frequent dosage requirements. Nystatin effectively targets Candida species by disrupting cell membranes, but formulation issues hinder clinical use. Lipid-based vesicular carriers, or novasomes, provide controlled, prolonged drug release and enhanced skin penetration. This study focuses on developing nystatin-loaded novasomal gels as an advanced drug delivery system to enhance therapeutic efficacy, bioavailability, and patient compliance. The formulation was prepared using a modified ethanol injection technique, combining stearic acid, oleic acid, Span 60, cholesterol, and Carbopol to produce a stable transdermal gel. Comprehensive in vitro characterization using FTIR, SEM, XRD, and thermal analysis confirmed the chemical compatibility, morphological uniformity, and physical stability of the nystatin-loaded novasomal gel. Entrapment efficiency differed significantly among the formulations (p < 0.05), with F7 achieving the highest value (80%). All formulations maintained pH levels within the skin-friendly range of 5.5 to 7.0. Viscosity measurements, ranging from 3900 ± 110 to 4510 ± 105 cP, confirmed their appropriate consistency for dermal use. Rheological analysis showed a dominant elastic response, as indicated by storage modulus values consistently higher than the loss modulus. Particle size ranged from 4143 to 9570 nm, while PDI values remained below 0.3, reflecting uniform particle distribution. Zeta potential values were strongly negative, supporting physical stability. XRD studies indicated reduced crystallinity of nystatin within the formulations, while FTIR confirmed drug-excipient compatibility. SEM images showed spherical particles within the micrometer range. In vitro release studies demonstrated sustained drug release over 12 h, with F6 releasing the highest amount. The novasomal gel formulations-maintained stability for 30 days, with no notable alterations in pH, viscosity, or entrapment efficiency. Antifungal evaluation showed a larger inhibition zone (23 ± 2 mm) compared with the plain drug solution (15 ± 1.6 mm), while the MIC value was reduced (4.57 µg/mL), indicating greater potency. Skin irritation assessment in rats revealed only minor, temporary erythema, and the calculated Primary Irritation Index (0.22) confirmed a non-irritant profile. These findings suggest that the developed novasomal gel offers a promising approach for enhancing the treatment of fungal infections by enabling prolonged drug release, minimizing dosing frequency, and improving patient compliance. Full article

Accurate and efficient short-term load forecasting is crucial for the secure and stable operation and scheduling of power grids. Addressing the inability of traditional Transformer-based prediction models to capture symmetric correlations between different feature sequences and their susceptibility to multi-scale feature influences, this paper proposes a short-term power distribution network load forecasting model based on an enhanced Komodo Mlipir Algorithm (KMA)—Multivariate Variational Mode Decomposition (MVMD)-Crossformer. Initially, the KMA is enhanced with chaotic mapping and temporal variation inertia weighting, which strengthens the symmetric exploration of the solution space. This enhanced KMA is integrated into the parameter optimization of the MVMD algorithm, facilitating the decomposition of distribution network load sequences into multiple Intrinsic Mode Function (IMF) components with symmetric periodic characteristics across different time scales. Subsequently, the Multi-variable Rapid Maximum Information Coefficient (MVRapidMIC) algorithm is employed to extract features with strong symmetric correlations to the load from weather and date characteristics, reducing redundancy while preserving key symmetric associations. Finally, a power distribution network short-term load forecasting model based on the Crossformer is constructed. Through the symmetric Dimension Segmentation (DSW) embedding layer and the Two-Stage Attention (TSA) mechanism layer with bidirectional symmetric correlation capture, the model effectively captures symmetric dependencies between different feature sequences, leading to the final load prediction outcome. Experimental results on the real power distribution network dataset show that: the Root Mean Square Error (RMSE) of the proposed model is as low as 14.7597 MW, the Mean Absolute Error (MAE) is 13.9728 MW, the Mean Absolute Percentage Error (MAPE) reaches 4.89%, and the coefficient of determination (R²) is as high as 0.9942. Full article

Background: Nephrolepis exaltata (sword fern) possesses a considerable amount of phytochemicals and different biological activities. The current study investigates the anti-biofilm potential of greenly synthesized bimetallic nanoparticles of Nephrolepis exaltata leaf methanol extract (NEME-MnO₂-MgO BNPs). Methods: The NEME was subjected to UPLC/MS analysis, followed by characterization of its NPs by size, zeta potential, FTIR, entrapment efficiency, and release. Then, antioxidant, antimicrobial and antibiofilm assays were employed, followed by in silico studies. Results: The UPLC/MS analysis of NEME led to the tentative identification of 27 metabolites, mostly phenolics. The MnO₂-MgO BNPs presented a uniform size and distribution and exhibited IC₅₀ values of 350 and 215.6 μg/mL, in the DPPH and ABTS assays, respectively. Moreover, the NPs exhibited antimicrobial and anti-biofilm efficacies against Pseudomonas aeruginosa, Klebsiella pneumonia (ATCC-9633), Staphylococcus aureus (ATCC-6538), Escherichia coli, Bacillus cereus, and C. albicans, with MIC values of 250–500 μg/mL. The MnO₂-MgO BNPs inhibited Candida albicans biofilms with a % inhibition of 66.83 ± 2.45% at 1/2 MIC. The network pharmacology highlighted epigallocatechin and hyperoside to be the major compounds responsible for the anti-biofilm potential. The ASKCOS facilitated the prediction of the redox transformations that occurred in the green synthesis, while the docking analysis revealed enhanced binding affinities of the oxidized forms of both compounds towards the outer membrane porin OprD of P. aeruginosa, with binding scores of −4.6547 and −5.7701 kcal/mol., respectively. Conclusions: The greenly synthesized Nephrolepis exaltata bimetallic nanoparticles may provide a promising, eco-friendly, and sustainable source for antimicrobial agents of natural origin with potential biofilm inhibition. Full article

Many scholars are interested in modeling complex data in an effort to create novel probability distributions. This article proposes a novel class of distributions based on the inverse of the exponentiated Weibull hazard rate function. A particular member of this class, the Weibull–Rayleigh distribution (WR), is presented with focus. The WR features diverse probability density functions, including symmetric, right-skewed, left-skewed, and the inverse J-shaped distribution which is flexible in modeling lifetime and systems data. Several significant statistical features of the suggested WR are examined, covering the quantile, moments, characteristic function, probability weighted moment, order statistics, and entropy measures. The model accuracy was verified through Monte Carlo simulations of five different statistical estimation methods. The significance of WR is demonstrated with three real-world data sets, revealing a higher goodness of fit compared to other competing models. Additionally, the change point for the WR model is illustrated using the modified information criterion (MIC) to identify changes in the structures of these data. The MIC and curve analysis captured a potential change point, supporting and proving the effectiveness of WR distribution in describing transitions. Full article

This study aimed to develop and characterise novel hydrogels based on natural bioactive compounds for topical antimicrobial applications. Four gel systems were formulated using different polymers, namely polyacrylic acid (Carbopol 940, CBP-G), chitosan with high and medium molecular weights (CTH-G and CTM-G), and sodium alginate (ALG-G), incorporating tinctures of Verbena officinalis and Aloysia triphylla, Laurus nobilis essential oil, and a β-cyclodextrin–clove essential oil complex. All gels displayed a homogeneous macroscopic appearance and maintained stability for over 90 days. Rheological studies demonstrated gel-like behaviour for CBP-G and ALG-G, with well-defined linear viscoelastic regions and distinct yield points, while CTM-G exhibited viscoelastic liquid-like properties. SEM imaging confirmed uniform and continuous matrices, supporting controlled active compound distribution. Thermogravimetric analysis (TG-DTA) revealed a two-step degradation profile for all gels, characterised by high thermal stability up to 230 °C and near-total decomposition by 500 °C. FTIR spectra confirmed the incorporation of bioactive compounds and products and highlighted varying interaction strengths with polymer matrices, which were stronger in CBP-G and CTH-G. Antimicrobial evaluation demonstrated that chitosan-based gels exhibited the most potent inhibitory and antibiofilm effects (MIC = 2.34 mg/mL) and a cytocompatibility assessment on HaCaT keratinocytes showed enhanced cell viability for chitosan gels and dose-dependent cytotoxicity for alginate formulations at high concentrations. Overall, chitosan-based gels displayed the most favourable combination of stability, antimicrobial activity, and biocompatibility, suggesting their potential for topical pharmaceutical use. Full article

Introduction: Staphylococcus aureus is a high-priority foodborne pathogen contributing to several food poisoning outbreaks. Methicillin- and vancomycin-resistant S. aureus (MRSA and VRSA), pose significant public health concerns due to their potential for serious illness, antibiotic resistance, and transmission within both healthcare and community settings. These bacteria can cause numerous infections, ranging from skin and soft tissue infections to life-threatening conditions like bloodstream infections, pneumonia, and endocarditis. Although several publications are concerned with Staphylococcus aureus contamination in ready-to-eat (RTE) food products, little published data is available about its prevalence in pizza, which is widely distributed and consumed worldwide. Methods: The current study is intended to determine the prevalence, virulence genes, and antimicrobial resistance profiles of S. aureus in three hundred ready-to-eat pizza samples (100 each of meat, chicken, and canned tuna pizzas) collected from different restaurants in Mansoura City, Egypt. The typical colonies on Baird–Parker selective agar supplemented with egg yolk tellurite emulsion were counted and further confirmed based on Gram staining, coagulase testing, catalase testing, carbohydrate fermentation, and thermostable nuclease production. The genomic DNA of the confirmed coagulase-positive isolates was prepared and subjected to PCR analyses for detecting the nuc gene, mecA (methicillin resistance gene), and vancomycin resistance gene (vanA), as well as six selected S. aureus virulence genes: sea, seb, sec, sed, hla, and tsst. The antimicrobial resistance profile of the S. aureus isolates was determined against 16 antimicrobial agents belonging to six classes using the agar disc diffusion method according to the Clinical and Laboratory Standards Institute guidelines (CLSI), except for oxacillin and vancomycin, which were assessed using the MIC test. Results: The results revealed that 56% (56/100), 56% (56/100), and 40% (40/100) of chicken, meat, and canned tuna pizzas were positive for S. aureus, with an overall prevalence of 50.7% (152/300). All 560 isolates (100%) were verified as S. aureus based on molecular confirmation of the nuc gene. Interestingly, 48.6% (272/560) and 8.6% (48/560) of the isolates tested were identified as methicillin- and vancomycin-resistant S. aureus (MRSA and VRSA) through detection of mecA and vanA genes, respectively. Among the S. aureus isolates tested, the hla gene was detected in 87.1% (488/560), while the enterotoxin genes sea, seb, sec, and sed were identified in 50% (280/560), 78.6% (440/560), 9.8% (55/560), and 24.5% (137/560) of isolates, respectively. All recovered isolates (n = 560) were classified as multidrug-resistant and were resistant to penicillin, oxacillin, and ampicillin. Moreover, 77% (431/560), 24% (134/560), 8% (45/560), and 8.6% (48/560) of isolates were resistant to cefotaxime, ciprofloxacin, azithromycin, and vancomycin, respectively. Conclusions: The current study emphasizes that ready-to-eat pizza is highly contaminated with multidrug-resistant S. aureus, highlighting the urgent need for rationalizing antibiotic use in both veterinary and human medicine to prevent the transmission of resistant bacteria through the food chain. Additionally, strict adherence to good hygienic practices throughout all stages of the food chain is essential to minimize overall contamination and enhance food safety. Full article

Background/Objectives: Current dosing recommendations for piperacillin/tazobactam suggest adjustments only for patients with creatinine clearance (CrCl) below 40 mL/min, potentially neglecting the variability in drug exposure among patients with a CrCl greater than 40 mL/min. This study aimed to develop a population pharmacokinetic (PK) model for piperacillin/tazobactam and explore optimal dosage regimens tailored by renal function and pathogen susceptibility. Methods: Twelve healthy adults received a single intravenous dose of piperacillin/tazobactam (4 g/0.5 g). Population PK models were developed using nonlinear mixed-effects modeling. Monte Carlo simulations were conducted to identify optimal dosing regimens across various renal functions and MIC levels, guided by pharmacodynamic targets defined as the percentage of time that free drug concentrations exceed the minimum inhibitory concentration (fT_>MIC). Results: PK profiles of both drugs were best described by two-compartment models. Estimated glomerular filtration rate (eGFR) adjusted by body surface area and body weight were identified as significant covariates influencing drug clearance and peripheral volume of distribution. Simulations showed that the standard dosing regimen (4/0.5 g q6h with 30 min infusion) achieved a 90% probability of target attainment (PTA) for 50%fT_>MIC at MIC values up to 4 mg/L in patients with normal renal function. However, this regimen often did not achieve a 90% PTA for stringent targets (100%fT_>MIC, 100%fT_>4MIC) or higher MICs, particularly in patients with eGFR ≥ 130 mL/min. Conclusions: These findings suggest current dosing regimens may be inadequate and highlight the potential of alternative strategies, such as extended or continuous infusion, which warrant further investigation in clinical populations to optimize therapeutic outcomes. Full article

Among synanthropic species, European hedgehogs are widely distributed throughout Europe. In recent decades, these animals have increasingly adapted to anthropogenic environments, where they find abundant shelter and food resources, along with fewer natural predators. As with other wildlife, it is likely that their coexistence in cities is also affecting their microbiota, promoting the development of antimicrobial resistance (AMR). This study aimed to assess the occurrence and patterns of AMR in commensal enteric Escherichia coli isolated from hedgehogs (n = 53) living in anthropogenic environments upon admission to a wildlife rescue center in Turin (Italy). The effects of hospitalization on the prevalence and trends of AMR were also assessed. Our results confirm that hedgehogs can harbor resistant E. coli upon admission, in particular against cefazolin (41.5%), ampicillin (37.7%), and enrofloxacin (22.6%). In addition, hospitalization promoted an increase in minimum inhibitory concentration (MIC) values of all antibiotics except imipenem, which led to a significant increase in E. coli that was resistant towards doxycycline, enrofloxacin, and trimethoprim-sulfamethoxazole. Admitted hedgehogs were also carriers of extended-spectrum beta-lactamase-producing E. coli (5.7%), whose presence increased during hospitalization (to 20.8%). These results highlight the role of hospitalizations longer than five days in the acquisition of AMR and suggest that European hedgehogs can become potential carriers of resistant E. coli following hospitalization. Full article

Rheum cordatum Losinsk is a plant species distributed in Kazakhstan but remains relatively understudied despite its promising biological potential. The present study aimed to explore leaf extracts of R. cordatum by utilizing advanced green extraction technologies including supercritical CO₂ (ScCO₂), subcritical ethanol (Sc) and ultrasound-assisted extraction (UAE) to characterize their phytochemical composition and evaluate their antioxidant and antimicrobial activities. A total of 53 phytochemical compounds were identified, with gallic acid (30.71 µg/mg UAE-EtOH-4h), rutin (21.93 µg/mg ScCO₂-150) and hesperidin (14.98 µg/mg ScCO₂-150) being notably abundant. Among the tested extracts, ScCO₂ extraction at 150 bar (ScCO₂-150) demonstrated the highest antioxidant activity, exhibiting IC₅₀ values of 0.0132 mg/mL (DPPH) and 0.0462 mg/mL (ABTS), coupled with the highest total phenolic content (140 mg GAE/g). Moreover, the ScCO₂-150 extract showed pronounced antimicrobial efficacy, particularly against Bacillus subtilis Pseudomonas aeruginosa and Staphylococcus aureus, with minimum inhibitory concentrations (MIC) ranging from 125 to 250 µg/mL. These findings highlight the considerable potential of R. cordatum leaves as a valuable, abundant and sustainable source of natural antioxidants and antimicrobial agents, with supercritical CO₂ extraction presenting substantial advantages in selectively obtaining bioactive phytochemicals. Full article

Asphalt pavement cracking is an important factor affecting its service life. Under certain conditions, the self-healing behavior of asphalt itself can repair pavement cracks. However, the self-healing ability of asphalt itself is limited. In order to strengthen the self-healing ability of asphalt, the microcapsule wrapped with a repair agent is pre-mixed into the asphalt mixture. When the crack occurs and spreads to the surface of the microcapsule, the microcapsule ruptures and the healing agent flows out to realize the self-healing of the crack. Current microcapsules are mostly prepared with healing agents and bio-oil as core materials, and their high-temperature resistance to rutting is poor. While the epoxy resin contains a three-membered cyclic ether, it can undergo ring-opening polymerization to bond and repair the asphalt matrix. In addition, research on microcapsules mainly focuses on the self-healing properties of microcapsule-modified asphalt. In fact, before adding microcapsules to asphalt to improve its self-healing performance, it is necessary to ensure that the asphalt has a good road performance. On this basis, the self-healing performance of asphalt is improved, thereby extending the service life of asphalt pavement. Therefore, two-component epoxy self-healing microcapsules (E-mic and G-mic) were first prepared in this paper. Then, a temperature scanning test, rheological test of bending beams, and linear amplitude scanning test were, respectively, conducted for the microcapsule/asphalt to evaluate its road performance, including the high-temperature performance, low-temperature crack resistance, and fatigue performance. Finally, the self-healing performance of microcapsules/asphalt was tested. The results showed that the self-developed epoxy self-healing microcapsules were well encapsulated and presented as spherical micron-sized particles. The average particle size of the E-mic was approximately 23.582 μm, while the average particle size of the G-mic was approximately 22.440 μm, exhibiting a good normal distribution. In addition, they can remain intact and unbroken under high-temperature conditions. The results of road performance tests indicated that the microcapsule/asphalt mixture exhibits an excellent high-temperature resistance to permanent deformation, low-temperature crack resistance, and fatigue resistance. The self-healing test demonstrated that the microcapsule/asphalt exhibited an excellent self-healing performance. When the microcapsule content was 4%, the self-healing rate reached its optimal level of 67.8%, which was 149.2% higher than that of the base asphalt. Full article

This study examined the eco-friendly synthesis of zinc oxide (ZnO) nanoparticles using Cladophora glomerata extracts as reducing and stabilizing agents, comparing zinc acetate and zinc chloride precursors for biomedical and environmental applications. Zinc acetate-synthesized ZnO nanoparticles showed a significant absorption peak around 320–330 nm, indicating stable, quasi-spherical ZnO nanoparticles with a narrow size distribution, primarily around 100 nm. Zeta potential measurements revealed a value of −25 mV for these particles, suggesting moderate colloidal stability. XRD analysis confirmed a highly crystalline hexagonal wurtzite structure for zinc acetate-derived ZnO, and SEM images supported a proper microstructure with approximately 2 µm particle size. FTIR analysis indicated higher-quality ZnO from zinc acetate due to the absence of moisture and hydroxyl groups. Conversely, zinc chloride-derived ZnO particles displayed a broader absorption spectrum around 370 nm, indicative of significant aggregation. Their narrower zeta potential distribution around +10 mV suggested diminished colloidal stability and a heightened aggregation tendency. While a peak around 100 nm was observed, many particles exceeded 1000 nm, reaching up to 10,000 nm. XRD results showed that zinc chloride adversely affected crystallinity, and SEM analysis indicated smaller particles (approx. 1 µm). FTIR analysis demonstrated that zinc chloride samples retained hydroxyl groups. Both zinc acetate- and zinc chloride-derived ZnO nanoparticles produced notable inhibitory zones against Gram-positive (L. monocytogenes, S. aureus) and specific Gram-negative bacteria (E. coli, K. pneumoniae). Zinc acetate-derived ZnO showed a 21 mm inhibitory zone against P. vulgaris, while zinc chloride-derived ZnO showed a 10.1 mm inhibitory zone against C. albicans. Notably, zinc chloride-derived ZnO exhibited broad-spectrum antimicrobial activity. MIC readings indicated that zinc acetate-derived ZnO had better antibacterial properties at lower concentrations, such as 3.125 µg/mL against L. monocytogenes. These findings emphasize that the precursor material selection critically influences particle characteristics, including optical properties, surface charge, and colloidal stability. Full article

Daucus is a large genus of the Apiaceae family, comprising around forty-five accepted species, that has a worldwide distribution. Species of this genus have been reported to have several traditional medicinal uses, and some of them are also largely used as food and spices. Daucus nebrodensis Strobl. is an endemic species of Sicily growing in the montane environments of the Madonie and the Nebrodi Mountains. In this work, the essential oil of D. nebrodensis (DnEO), collected wild near Messina (Italy), was chemically and biologically investigated. The hydrodistilled essential oil (yield 0.15%), obtained from fresh aerial parts, was evaluated by GC-MS, and It was particularly rich in monoterpene hydrocarbons, with sabinene (33.6%), α-pinene (17.2%), γ-terpinene (9.8%), and α-terpinene (7.6%) as the main metabolites. DnEO, and its main constituents, have been tested to evaluate their biological properties. Given the current problem of antibiotic resistance, it is of great interest to identify alternative molecules that could counteract the its progression. Therefore, DnEO was tested against Gram-negative species, such as E. coli DH5α and P. aeruginosa PAOI, and Gram-positive species, such as S. aureus ATCC6538P, B. subtilis AZ54, and M. smegmatis MC²155, showing notable antibacterial activity. The MIC for Bacillus subtilis, the most sensitive strain, was 18 mg/mL, while the MIC for Pseudomonas aeruginosa, the least sensitive strain, was 30 mg/mL. Moreover, interesting antibiofilm activity was observed against Mycobacterium smegmatis with a 55% inhibition. Its ability to form biofilms contributes to its persistence and resistance in clinical settings. These findings highlight the potential of D. nebrodensis EO as a source of bioactive compounds with promising antimicrobial and antibiofilm properties. Full article

This research presents a novel, sustainable, and eco-friendly method for the rapid green synthesis of nanoparticles with antibacterial properties. This method employs steam distillation to extract reducing and stabilizing agents from orange peel waste, followed by ultrasound-assisted synthesis. To the best of our knowledge, this is the first reported integration of these two techniques for nanoparticle production. The extracted materials were then subjected to rigorous characterization through a combination of analytical techniques, including FTIR, HPLC, and TEM. These analytical approaches enabled a comprehensive analysis of the synthesized NPs, revealing their size distribution within the range of 1.5 to 14 nm. Among the synthesized nanomaterials, AgNPs exhibited the most potent antibacterial activity, with statistically significant minimum inhibitory concentrations (MICs) of 16 ppm for E. coli ATCC and 32 ppm for resistant E. coli and E. faecalis strains. This study underscored the promise of valorizing citrus waste for nanomaterial synthesis and introduced a novel, scalable methodology for producing bioactive nanoparticles, promoting a more sustainable technology for this purpose. Notably, this research aligns with United Nations Sustainable Development Goal 12, which promotes responsible consumption and production by transforming organic waste into high-value functional nanomaterials for biomedical and environmental applications. Full article

The increasing emergence of antifungal resistance poses potential clinical challenges, particularly among immunocompromised patients with cancer at risk of invasive mold infections, but data on antifungal susceptibility trends specific to this population are few. We evaluated distributions of minimal inhibitory concentrations (MIC), including minimal effective concentrations (MEC) for echinocandins, of 11 antifungal agents for 523 mold isolates (395 Aspergillus spp.) from cancer patients. Based on published Clinical and Laboratory Standards Institute guidelines, isavuconazole had notably high rates of non-wild-type MICs for A. fumigatus (19.6%), A. flavus/oryzae (34.8%), A. niger complex (26.1%), and A. terreus complex (8.33%). Persistent low baseline resistance of A. fumigatus to voriconazole was observed across multiple years (2.4–11.5% per year, average 8.41%) without significant trends in MIC change over time. Itraconazole and posaconazole demonstrated the lowest MIC distributions (MIC₅₀ ≤ 0.06–0.5 µg/mL) of the azoles against Aspergillus spp. Amongst the A. niger complex, 29.4% (27/92) demonstrated non-wild-type MICs to itraconazole. While the A. nidulans group was less frequent (n = 24), bimodal peaks in MIC/MEC were noted for caspofungin (≤0.06 and 1 µg/mL). Non-Aspergillus molds of significance (Zygomycetes, Fusarium spp., Scedosporium spp., and Lomentospora prolificans) demonstrated variable but increased MICs to antifungal agents as previously described. Our results highlight increased rates of non-wild type MICs for Aspergillus spp. to isavuconazole and voriconazole, which are commonly used antifungal agents in cancer patients. Such AST trends should be closely monitored in populations with frequent antifungal use and encourage increased antifungal stewardship efforts. Full article

Antimicrobial resistance is a threat to probiotic microorganisms due to their potential role in harboring and transmitting resistance genes. This study focuses on two Bifidobacterium species (B. animalis subsp. lactis and B. longum) by analyzing 657 Minimal Inhibitory Concentration (MIC) values extracted from research articles indexed in Scopus, PubMed, and Web of Science, published since 2014, and considering 17 different antibiotics. MIC values were used for descriptive statistical analysis (boxplots and violin plots) to evaluate both inter- and intraspecies distributions. The results showed an overall increase in MIC values compared to historical data, with B. longum exhibiting high resistance to tetracyclines and streptomycin—approximately 25% to 50% of the strains had MIC values > EFSA cut-offs. The violin plots revealed the presence of resistant subpopulations, particularly within B. longum. These findings support the relevance of longitudinal MIC analysis as a tool for detecting early shifts in antimicrobial susceptibility and highlight the importance of data-driven approaches for microbiological risk assessment in probiotic applications. Full article