Search Results (1,368)

Background/Objectives: Staphylococcus sciuri, traditionally regarded as a commensal organism in animals and the environment, is increasingly recognized as a potential opportunistic pathogen with zoonotic significance. Its genomic reservoir of methicillin resistance homologues further raises concern regarding its role in antimicrobial resistance dissemination. This study describes the first documented case of S. sciuri isolated from the urinary bladder of a domestic rabbit (Oryctolagus cuniculus) in Romania, emphasizing its clinical relevance and antimicrobial profile. Methods: A seven-year-old intact female rabbit presenting with apathy, dysuria, and hematuria underwent clinical evaluation, ultrasonography, and cystocentesis. The aspirated intravesical content was subjected to bacterial culture, MALDI-TOF MS identification, and antimicrobial susceptibility testing via the VITEK 2 system. Results: Pure colonies of Gram-positive cocci were identified as S. sciuri with high confidence. Antimicrobial susceptibility testing revealed susceptibility to β-lactams, aminoglycosides, glycopeptides, linezolid, rifampicin, fusidic acid, tigecycline, and trimethoprim-sulfamethoxazole, while resistance was observed against fluoroquinolones, macrolides, lincosamides, and tetracycline, indicating a multidrug-resistant phenotype. Treatment with trimethoprim-sulfamethoxazole combined with ultrasound-guided bladder lavage and supportive therapy resulted in complete clinical recovery within 10 days. Conclusions: This case highlights the pathogenic potential of S. sciuri in domestic rabbits and its capacity to exhibit multidrug resistance. The findings underscore the necessity of including rabbits in antimicrobial resistance surveillance programs and reinforce the importance of culture and sensitivity testing in guiding the therapeutic management of exotic companion animals. Full article

In this study, Enterococcus faecalis strains isolated from raw cow’s milk were examined for genetic diversity, ability to produce biogenic amines (including histamine, tyramine, putrescine, cadaverine, 2-phenylethylamine) and the presence of corresponding amino acid decarboxylase genes. Identification of 29 strains obtained from Polish farms was carried out by polymerase chain reaction (PCR) and matrix-assisted laser desorption and ionization time of flight (MALDI-TOF MS) methods, and their genetic relationships were assessed by the Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) technique. Amine production capacity was assessed in vitro on synthetic medium, while the presence of decarboxylase genes (hdcA, tyrS, tyrDC, Odc, ldc) was detected by molecular assays, with the use of optimized primers enabling the detection of tyrDC in strains previously considered negative. The results showed high variability between strains and the ability of some isolates to produce high concentrations of tyrDC (max. > 1000 mg/kg); the presence of the tyrDC gene was strongly correlated with high production, although tyrDC-positive strains with low production were also reported, suggesting the influence of regulatory or environmental factors. The study underscores the need for precise molecular tools and systematic monitoring of biogenic amines to ensure the safety and quality of dairy products. Full article

Bacterial outer-membrane vesicles (OMVs) mediate stress tolerance, biofilm formation, and interkingdom communication, but their role in beneficial endophytes remains underexplored. We isolated 11 non-redundant isolates associated with Bacillus, Enterococcus, Kosakonia and Kocuria from Agave tequilana seeds, identified by MALDI-TOF MS and 16S rRNA gene sequencing. We focused on the catalase-negative Enterobacter cloacae SEA01, which exhibits plant-promoting traits and support agave growth under nutrient-poor microcosms. In addition, this endophyte produces OMVs. Time-resolved SEM documented OMV release and cell aggregation within 9 h, followed by mature biofilms at 24 h with continued vesiculation. Purified OMVs (≈80–300 nm) contained extracellular DNA and were characterized by dynamic light scattering and UHPLC–ESI–QTOF-MS lipidomics. The OMV lipidome was dominated by phosphatidylethanolamine (~80%) and was enriched in monounsaturated fatty acids (16:1, 18:1), while the stress-associated cyclopropane fatty acids (17:1, 19:1) were comparatively retained in the whole-cell membranes; OMVs also exhibited reduced ubiquinone-8. SEA01 is catalase-negative, uncommon among plant-associated Enterobacter, suggesting a testable model in which oxidative factors modulate OMV output and biofilm assembly. These may have implications for recognition and redox signaling at the root interface. Future works should combine targeted proteomics/genomics with genetic or chemical disruption of catalase/OMV pathways. Full article

Background/Objectives: Streptococcus uberis is a Gram-positive bacterium and a major cause of bovine mastitis. The use of antimicrobial treatments raises concerns about resistance. This study aimed to characterize S. uberis isolates from one of the ten largest milk-producing regions in Europe. Methods: Thirty-six isolates from 36 cows with mastitis were identified using MALDI-TOF and VITEK^®MS. Susceptibility to 9 antibiotics (penicillin G, ampicillin, tetracycline, erythromycin, clindamycin, cefotaxime, ceftriaxone, levofloxacin, and moxifloxacin) was determined with VITEK^®2. Whole-genome sequencing was performed using MinION Mk1C. Results: Alleles were identified for 7 loci: arcC, ddl, gki, recP, tdk, tpi, and yqiL. Only 10 isolates had alleles for all the loci. The loci with the highest number of alleles were ddl and tdk (33/36 strains), while arcC had the fewest (19/36). Four isolates were assigned to known sequence types (ST6, ST307, and ST184), and novel alleles were detected in 32 of the 36 isolates. Twelve isolates showed phenotypic resistance to one or more of the following antibiotics: tetracycline, erythromycin, clindamycin, and ceftriaxone. The lnu was the most frequently detected resistance gene (27 out of 102 total gene appearances). A total of 19 virulence factors were identified. All strains were predicted to be capable of infecting human hosts. Conclusions: Streptococcus uberis is a potential reservoir of antimicrobial resistance genes. The use of antimicrobials to treat bovine mastitis has reduced the susceptibility of this microorganism to several antibiotics, underscoring the importance of monitoring antimicrobial use in veterinary practice. The results also highlight the high genetic diversity of the isolates, suggesting a strong capacity to adapt to different environmental conditions. Full article

Background: Land use change fundamentally alters soil microbial communities and biochemical processes, yet the integrated effects on rhizosphere microbiome–metabolome networks remained poorly understood. Objective: This study investigated land uses as forest, grassland and intermediary edge shape the rhizosphere biochemical networks of naturally grown Barbarea vulgaris. Methods: Rhizosphere soils of Barbarea vulgaris were analysed for microbial community structure abundance, and metabolomic profile applying phospholipid fatty acid (PLFA) profiling and mass spectrometric untargeted metabolomics (GC–MS/MS and MALDI–TOF/TOF MS). These were coupled with co–inertia analysis to assess microbiome–metabolome interactions. Results: Microbial community analysis revealed significant effects of land use on bacterial community structure (G+/G−, p < 0.001). Untargeted metabolomics identified 248 metabolites, of which 161 were mapped to KEGG pathways. Amino acids and derivatives (21.1%) followed by organic acids (16.8%) were the most representative among identified metabolites. Pathway enrichment analysis revealed coordinated reprogramming of central carbon and nitrogen metabolism across land use gradients, particularly in the amino acid metabolism, TCA cycle, and glycolysis/gluconeogenesis pathways. Microbiome–metabolome coupling analysis revealed distinct correlation patterns between microbial phenotypes and metabolite classes, with forest environments showing the strongest biochemical network integration (RV = 0.91). Edge habitats presented intermediate signatures, supporting their role as transitional zones with unique biochemical properties. Conclusions: The environmental context fundamentally shapes rhizosphere biochemical network organization through coordinated shifts in bacterial community structure and metabolic pathway activity. These habitat-specific metabolic signatures suggest that land use change triggers adaptive biochemical responses that may influence plant performance and ecosystem functioning across environmental gradients. Full article

Despite the devastating consequences of fungal disease, research struggles to catch up to present needs. This study aims to give a broad perspective on the situation, investigating patterns and distribution of fungal pathogens and monitoring trends of resistance to antifungal drugs, over an 8-year timeframe, at the National Institute of Infectious Diseases “Prof. Dr. Matei Balș” in Bucharest, Romania. Samples were inoculated on Sabouraud or Brilliance Candida Agar media; strains were identified using MALDI-TOF MS; and antifungal sensitivity testing was performed using E-Tests strips, VITEK2 Compact and MICRONAUT-AM automatic systems. Candida albicans, accounting for 42% of the positive samples, was the most common pathogen observed, with only 17% of the isolates being susceptible to all antifungals tested, while it was also predominant and deadly in the ICU. The emerging Candidozyma auris, found in 8% of the candidoses, exhibited a fluconazole resistance rate of 96.3%. Of the Aspergillus fumigatus strains, 35.7% showed resistance to azoles, and 25% to amphotericin B. In the ICU, more than half of A. flavus-, A. fumigatus- or A. niger-related cases culminated in death. Antifungal resistance is not to be treated lightly, as it is still a complex and dynamic threat, with devastating consequences. Full article

Preventive immunology is emerging as a cornerstone of animal infectious disease control within One Health, shifting emphasis from treatment to prevention. This review integrates mechanistic insights in host immunity with a comparative evaluation of next-generation interventions—mRNA/DNA and viral-vector vaccines, nanovaccines, monoclonal antibodies, cytokine modulators, probiotics/postbiotics, bacteriophages, and CRISPR-based approaches—highlighting their immunogenicity, thermostability, delivery, and field readiness. Distinct from prior reviews, we appraise diagnostics as preventive tools (point-of-care assays, biosensors, MALDI-TOF MS, AI-enabled analytics) that enable early detection, risk prediction, and targeted interventions, and we map quantifiable links between successful prevention and reduced antimicrobial use. We embed translation factors—regulatory alignment, scalable manufacturing, workforce capacity, equitable access in LMICs, and public trust—alongside environmental and zoonotic interfaces that shape antimicrobial resistance dynamics. We also provide a critical analysis of limitations and failure cases: gene editing may require stacked edits and concurrent vaccination; phage programs must manage host range, resistance, stability, and regulation; and probiotic benefits remain context-specific. Finally, we present a risk–benefit–readiness framework and a time-bound research agenda to guide deployment and evaluation across animal–human–environmental systems. Coordinating scientific innovation with governance and ethics can measurably reduce disease burden, curb antimicrobial consumption, and improve health outcomes across species. Full article

Introduction: Klebsiella variicola, a member of Klebsiella pneumoniae complex, has emerged as an opportunistic pathogen for human infection; however, antimicrobial resistance and hypervirulent characteristics of K. variicola have rarely been investigated in South Korea. Methods: We analyzed 76 clinical K. variicola isolates collected from 12 hospitals between September 2022 and October 2023. Bacterial identification was performed by MALDI-TOF MS. Antimicrobial susceptibility was tested by disk diffusion tests. Resistance determinants and virulence traits were investigated, and whole-genome sequencing was performed for hypermucoviscous or carbapenem-resistant K. variicola isolates. Results: Most (89.5%, 68/76) were susceptible to all 18 antimicrobials tested in this study, and 3 isolates harbored bla_CTX-M-15. One isolate carried bla_KPC-2 on its IncX3 plasmid, which is closely related to carbapenem-resistant K. pneumoniae plasmids. Capsular typing revealed 51 wzi allelic types. Ten isolates showed mucoid phenotype, mainly with KL60 and KL61. Conclusions: This study reveals relatively low resistance rates in K. variicola strains but the presence of multidrug-resistant and hypermucoviscous K. variicola strains. In addition, the evidence of interspecies dissemination of bla_KPC-2 highlights the need for continuous genomic surveillance. Full article

Ready-to-eat (RTE) foods can carry antimicrobial-resistant pathogens; however, few studies link real-world surveillance to practical interventions. This study addressed this gap by estimating the prevalence of Staphylococcus aureus (S. aureus) and methicillin-resistant S. aureus (MRSA) in ready-to-eat foods from Al-Qassim and evaluating a rapid, orthogonal confirmation workflow (culture → MALDI-TOF MS → Vitek 2 → mecA/mecC PCR). The in vitro activity of citrate-stabilized silver nanoparticles (AgNPs) against food-derived MRSA was quantified, and synergy with oxacillin (primary) and ciprofloxacin (secondary) was examined. Silver-susceptibility stability was assessed over 20 days of sub-MIC serial passage, with attention to whether β-lactam co-exposure constrained drift. We surveyed 149 RTE products and paired the confirmation workflow with mechanistic tests of AgNPs as antibiotic adjuvants. S. aureus was recovered from 24.2% of products and MRSA from 6.7%, with higher recovery from animal-source matrices and street-vendor outlets. MALDI-TOF MS provided rapid species confirmation and revealed two reproducible low-mass peaks (m/z 3990 and 4125) associated with MRSA, supporting spectral triage pending molecular confirmation. Antimicrobial susceptibility testing showed the expected β-lactam split (MRSA oxacillin/cefoxitin non-susceptible; MSSA oxacillin-susceptible but largely penicillin-resistant), with last-line agents retained. Citrate-stabilized AgNPs displayed consistent potency against food-derived MRSA (MIC 8–32 µg/mL; MIC₅₀ 16; MIC₉₀ 32) and were predominantly bactericidal (MBC/MIC ≤ 4 in 90%). Checkerboards demonstrated frequent AgNP–oxacillin synergy (median fractional inhibitory concentration index [FICI] 0.37; 4–16-fold oxacillin MIC reductions) and additive-to-synergistic effects with ciprofloxacin (median FICI 0.63), translating time–kill assays into rapid, sustained bactericidal activity without antagonism. During sub-MIC evolution, silver MICs rose modestly (median two-fold) and often regressed off drug; oxacillin co-exposure limited drift. RTE foods therefore represent credible MRSA exposure routes. Integrating MALDI-assisted triage with automated AST enables scalable surveillance, and standardized AgNP formulations emerge as promising β-lactam adjuvants—pending in situ efficacy, safety, and residue evaluation. Full article

Background: In animals, staphylococci constitute a significant part of the normal skin microbiota and mucous membranes. There is limited information available on staphylococci isolated from healthy horses. These skin-associated bacteria can be easily transferred between animals and horse riders via direct contact. Patients undergoing hippotherapy (i.e., medical or therapeutic sessions with horses) are especially at risk of being colonized by horse skin-associated bacteria. However, it remains unclear whether equine skin is colonized by antimicrobial-resistant (AMR) opportunistic pathogens, which may be of concern to human health. Methods: We cultivate staphylococci from samples collected from healthy, non-vet-visiting horses who live on private farms in a rural area. In total, 61 strains were isolated and identified at the species level using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). Results: The diversity of Staphylococcus species in the equine skin microbiota was relatively high and, with the exception of S. aureus, all the other recovered strains were coagulase-negative staphylococci (CoNS). In total, eleven different staphylococcal species were identified: S. xylosus, S. sciuri, S. vitulinus, S. equorum, S. succinus, S. nepalensis, S. lentus, S. fleurettii, S. aureus, S. chromogenes, and S. simulans. Conclusions: These results indicate that healthy equine skin is colonized by opportunistic pathogens that can be causative agents of infections that are also severe in humans. The resistance among the isolated strains was observed in eight antimicrobials of the total tested and 36% (22/61) of the isolates were resistant to at least one antimicrobial. However, their resistance to critically important antibiotics used in human medicine was low. Seven isolates (11.5%; 7/61) were classified as multidrug-resistant (MDR). S. aureus (1/61) showed MDR and was methicillin-resistant. The S. aureus isolate contained genes conferring resistance to antibiotics, i.e., β-lactams (blaZ, mecA), aminoglycosides (aac(6′)/aph(2″)), and macrolide–lincosamide–streptogramin B (erm(B), erm(C), and lun(A/B)). Also CoNS harbored genes conferring resistance to β-lactams (blaZ), aminoglycosides (aac(6′)/aph(2″), ant(4′)-Ia), MLSB (erm(B), erm(C), lun(A/B)), and tetracycline (tetK, tetL). Full article

Background/Objectives: Staphylococcus pseudintermedius (S. pseudintermedius) is an opportunistic pathogen frequently isolated from dogs, involved in a wide range of infections, particularly otitis externa. Increasing antimicrobial resistance (AMR), including methicillin-resistant S. pseudintermedius (MRSP), poses significant challenges for veterinary and potentially human health. This study aimed to assess the prevalence and antimicrobial resistance profiles of S. pseudintermedius in dogs with otitis externa compared to clinically healthy dogs. Methods: Between 2022 and 2025, samples were collected from 400 dogs with otitis externa and 360 healthy dogs in veterinary clinics from Timișoara. Ear swabs were processed by conventional microbiological techniques and confirmed using MALDI-TOF MS. Antimicrobial susceptibility was tested using the VITEK^® 2 Compact system, following CLSI VET01, Fifth Edition (2018) standards. Fourteen antimicrobials from 11 classes were evaluated. Results: S. pseudintermedius was isolated in 40% of dogs with otitis externa and in 21.1% of healthy dogs. The highest resistance in both groups was observed to tetracycline (37.5% and 25%, respectively). No resistance was recorded to linezolid, vancomycin, teicoplanin, tigecycline, or fusidic acid. MRSP strains were identified in 1.2% of dogs with otitis, displaying multidrug resistance (MDR). MDR strains were also detected in 8.7% of diseased and 4% of healthy dogs, indicating the potential for subclinical reservoirs. Conclusions: The findings highlight the notable prevalence and AMR of S. pseudintermedius in both symptomatic and asymptomatic dogs. The detection of MRSP and MDR strains emphasizes the need for prudent antibiotic use and continuous AMR surveillance in veterinary medicine to mitigate zoonotic risks and preserve antimicrobial efficacy. Full article

Photodynamic therapy (PDT) has evolved considerably over the past decades, progressing from first-generation porphyrins to second- and third-generation photosensitizers, including nanocarrier-based systems with improved selectivity and bioavailability. In parallel, matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) has become a gold standard for the characterisation of complex biomolecules, enabling precise determination of molecular mass, purity and stability. This narrative review explores the intersection of these two fields, focusing on how MALDI-TOF MS supports the development, characterisation and clinical application of photosensitizers used in PDT. Literature searches were performed across PubMed, Web of Science, Scopus and Base-search, followed by targeted retrieval of studies on MALDI and PDT applications. Findings indicate that MALDI-TOF MS plays a crucial role at multiple stages: confirming the synthesis and chemical integrity of novel photosensitizers, monitoring their metabolic stability in biological systems and characterising photodegradation products after PDT. Moreover, MALDI imaging mass spectrometry (MALDI-IMS) enables spatial mapping of photosensitizer distribution in tissues, while rapid pathogen identification by MALDI-TOF supports antimicrobial PDT applications. Collectively, the evidence highlights that MALDI-MS is not only a tool for molecular characterisation but also a versatile analytical platform with a direct translational impact on PDT. Its integration with other omics and multimodal imaging approaches is expected to enhance the personalization and clinical effectiveness of photodynamic therapy. Full article

Background: Urinary tract infections (UTIs) are among the most common hospital- and community-acquired infections, creating a substantial healthcare burden due to recurrence, complications, and rising antimicrobial resistance. Accurate diagnosis and timely antimicrobial therapy are essential. This study aimed to identify trends in the etiology, treatment, and resistance patterns of UTIs through a retrospective analysis of urine isolates processed at the Laboratory of Microbiology at University Hospital St. George in Plovdiv, the largest tertiary care and reference microbiology center in Bulgaria, between 2017 and 2022. Materials and Methods: A retrospective single-center study was performed at the hospital’s Microbiology Laboratory. During the study period, 74,417 urine samples from 25,087 hospitalized patients were screened with the HB&L UROQUATTRO system. Positive specimens were cultured on blood agar, Eosin-Methylene Blue, and chromogenic media. Identification was performed using biochemical assays, MALDI-TOF MS, and the Vitek 2 Compact system. Antimicrobial susceptibility testing included disk diffusion, MIC determination, broth microdilution (for colistin), and Vitek 2 Compact, interpreted according to EUCAST standards. Descriptive analysis and temporal resistance trends were evaluated with regression models, and interrupted time-series analysis was applied to assess COVID-19-related effects. Results: Out of 10,177 isolates, Gram-negative bacteria predominated (73%), with Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis as the leading pathogens. Among Gram-positives, Enterococcus faecalis was the most frequent. In the post-COVID-19 period, ESBL production increased in E. coli (34–38%), K. pneumoniae (66–77%), and P. mirabilis (13.5–24%). Carbapenem resistance rose in K. pneumoniae (to 40.6%) and P. aeruginosa (to 24%), while none was detected in E. coli. Colistin resistance increased in K. pneumoniae but remained absent in E. coli and P. aeruginosa. High-level aminoglycoside resistance in E. faecalis was stable (~70%), and vancomycin resistance in E. faecium rose from 4.6% to 8.9%. Conclusions: Both community- and hospital-acquired UTIs in Southeastern Bulgaria are increasingly linked to multidrug-resistant pathogens, particularly ESBL-producing and carbapenem-resistant Enterobacterales. Findings from the region’s largest referral center highlight the urgent need for continuous surveillance, rational antibiotic use, and novel therapeutic approaches. Full article

Background/Objectives: Streptococcus suis (SS), an important zoonotic pathogen, has caused significant economic losses to the global pig industry. Existing commercial vaccines for SS mainly provide effective protection against a single serotype. Due to the existence of many serotypes and their robust immune evasion capabilities, the development of multi-component subunit vaccines or multi-epitope vaccines that provide effective cross-protection against different strains of SS is a key focus of current research. Methods: We applied two-dimensional electrophoresis (2-DE) and immunoblotting to screen for candidate immunogens among the immunogenic cell wall proteins of SS. BALB/c mice were immunized intradermally with a multi-component, multi-epitope vaccine. The vaccine’s safety and immunogenicity were assessed via clinical monitoring, antibody titer detection, cytokine assays, and survival curve analyses. Results: In this study, eight immunogenic cell wall proteins (GH25, Pk, PdhA, Ldh, ExoA, Pgk, MalX, and Dnak) were successfully identified using MALDI-TOF-MS, all of which could induce high IgG antibody titers. Based on the conservation and immunoprotection demonstrated by these eight protective antigenic proteins, PdhA, Ldh, and MalX were screened to construct a multi-component subunit vaccine as a candidate vaccine for providing cross-protection against SS isolates of multiple serotypes. Challenge studies showed that mice immunized with the multi-component subunit vaccine (PdhA, Ldh, and MalX) were protected against challenges with the SS2 virulent strain ZY05719 (62.5% protection) and the SSChz virulent strain CZ130302 (75% protection). Subsequently, we utilized immunoinformatics techniques to design a novel multi-epitope vaccine (MVPLM) derived from the immunogenic proteins PdhA, Ldh, and MalX. However, challenge tests revealed that the MVPLM offered limited protection against SS. Conclusions: These data demonstrate that a multi-component subunit vaccine composed of PdhA, Ldh, and MalX proteins shows promise as a candidate universal vaccine against multiple SS serotypes. Full article

Trichophyton indotineae is an emerging dermatophyte species responsible for recalcitrant and terbinafine-resistant dermatophytosis, raising concerns over diagnostic accuracy and treatment efficacy. This study aimed to improve the identification and resistance profiling of T. indotineae by integrating molecular methods with machine learning-assisted analysis of MALDI-TOF mass spectra. A total of 56 clinical isolates within the Trichophyton mentagrophytes complex were analyzed using ITS and ERG1 gene sequencing, antifungal susceptibility testing, and MALDI-TOF MS profiling. Terbinafine resistance was detected in 23 isolates and correlated with specific ERG1 mutations, including F397L, L393S, F415C, and A448T. While conventional MALDI-TOF MS failed to reliably distinguish T. indotineae from closely related species, unsupervised statistical methods (PCA and hierarchical clustering) revealed distinct spectral groupings. Supervised machine learning algorithms, particularly PLS-DA and SVM, achieved 100% balanced accuracy in species classification using 10-fold cross-validation. Biomarker analysis identified discriminatory spectral peaks for both T. indotineae and T. mentagrophytes (3417.29 m/z and 3423.53 m/z). These results demonstrate that combining MALDI-TOF MS with multivariate analysis and machine learning improves diagnostic resolution and may offer a practical alternative to sequencing in resource-limited settings. This approach could enhance the routine detection of terbinafine-resistant T. indotineae and support more targeted antifungal therapy. Full article