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Food products that are ready-to-eat have become increasingly popular in recent years due to their efficiency, affordability, and convenience. However, there are concerns about public health because certain products, particularly animal products, may contain antibiotic-resistant bacteria. This study aimed to quickly and accurately identify foodborne pathogens, such as Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus), in samples of shawarma and chicken burgers using peptide mass fingerprinting (PMF) technology. Additionally, the prevalence and levels of antibiotic resistance in the pathogens were determined. The study utilized 300 samples obtained from fast food restaurants in Al Qassim, Saudi Arabia. A variety of methods were used to identify foodborne pathogens, including culture on specific media, bacterial counts by numerical dilutions of homogenized samples, and proteome identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The Kirby–Bauer method was applied to detect the susceptibility and resistance of the bacteria to various antibiotics. PCR was utilized to identify antimicrobial resistance genes such as blaTEM, tet(A), blaZ, and mecA in S. aureus and E. coli isolates. The percentage of E. coli, S. aureus, Salmonella, Listeria monocytogenes (L. monocytogenes), Acinetobacter baumannii (A. baumannii), and Hafnia alevei (H. alevei) was 34%, 31%, 10.67%, 7.33%, 6.67%, and 4%, respectively. Shawarma samples were found to contain the highest levels of pathogens, compared with chicken burger samples. According to the MBT Compass Flex Series Version 1.3 software, all isolates were identified with 100% accuracy. The log score for MBT identification ranged from 2.00 to 2.56. Among E. coli isolates, ampicillin, and penicillin had the highest resistance rate (100%), followed by tetracycline (35.29%). A number of antibiotics were reported to be resistant to S. aureus, including nalidixic acid (100%), followed by penicillin (96.77%), piperacillin (45.16%), and norfloxacin (32.26%). Some E. coli isolates were susceptible to tetracycline (49.02%), nalidixic acid (47.06%), and piperacillin (43.14%), whereas amikacin was the only drug that was effective against 32.72% of S. aureus isolates. The proportions of the blaTEM and tet(A) genes in E. coli isolates were 55.89% and 45.1%, respectively, whereas S. aureus strains did not possess either of these genes. However, 21.5% and 47.31% of blaz and mecA genes were present among various isolates of S. aureus, respectively. In contrast, E. coli strains did not possess either of these genes. In conclusion, the fast identification and antimicrobial profiles of the foodborne pathogens were useful in identifying which restaurants and fast food outlets may need to improve their food safety practices. Ultimately, our results will be used to devise targeted strategies to control foodborne pathogens.

With the global rise in antimicrobial resistance (AMR), rapid and reliable microbial diagnostics have become more critical than ever. Traditional culture-based and molecular diagnostic techniques often fall short in terms of speed, cost-efficiency, or scalability, particularly in resource-limited settings. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI–TOF MS) has emerged as a transformative tool in clinical microbiology. Its unparalleled speed and accuracy in microbial identification, along with expanding applications in AMR profiling, make it a leading candidate for next-generation diagnostic workflows. This review aims to provide a comprehensive update on recent advances in MALDI–TOF MS, focusing on its technological evolution, clinical applications, and future potential in microbial diagnostics and resistance detection. We conducted a critical synthesis of peer-reviewed literature published over the last decade, with emphasis on innovations in sample preparation, instrumentation, data interpretation, and clinical integration. Key developments in AMR detection, including growth-based assays, resistance biomarker profiling, and machine learning-driven spectral analysis, are discussed. MALDI–TOF MS is increasingly deployed not only in clinical laboratories but also in environmental surveillance, food safety, and military biodefense. Despite challenges such as database variability and limited access in low-income regions, it remains a cornerstone of modern microbial diagnostics and holds promise for future integration into global AMR surveillance systems.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) has become one of the most serious problems confronting modern healthcare, particularly in intensive care units where patients are highly susceptible, procedures are frequent, and antibiotic exposure is often prolonged. In this review, carbapenem resistance in K. pneumoniae is presented not as a fixed feature of individual bacteria, but as a process that is constantly changing and closely interconnected. We bring together evidence showing how the spread of successful bacterial lineages, the exchange of resistance genes, and gradual genetic adjustment combine to drive both the rapid spread and the long-lasting presence of resistance. A major focus is placed on mobile genetic elements, including commonly encountered plasmid backbones, transposons, and insertion sequences that carry carbapenemase genes such as blaKPC, blaNDM, and blaOXA-48-like. These elements allow resistance genes to move easily between bacteria and across different biological environments. The human gut plays a particularly important role in this process. Its microbial community serves as a largely unseen reservoir where resistance genes can circulate and accumulate well before infection becomes clinically apparent, making prevention and control more difficult. This review also discusses the key biological factors that shape resistance levels, including carbapenemase production, changes in the bacterial cell membrane, and systems that expel antibiotics from the cell, and explains how these features work together. Advances in molecular testing have made it possible to identify resistance more quickly, supporting earlier clinical decisions and infection control measures. Even so, current tests remain limited by narrow targets and may miss low-level carriage, hidden genetic reservoirs, or newly emerging resistance patterns. Finally, we look ahead to approaches that move beyond detection alone, emphasizing the need for integrated surveillance, thoughtful antibiotic use, and coordinated system-wide strategies to lessen the impact of CRKP.

Antimicrobial resistance (AMR) remains a major clinical challenge, with Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species (ESKAPE) accounting for a substantial share of multidrug-resistant (MDR) infections worldwide. These organisms undermine antibiotic efficacy through reduced permeability, surface shielding, biofilm formation, and rapid genetic adaptation, mechanisms that primarily restrict effective exposure at infection sites. Bacteriophages, phage-derived enzymes, and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based antimicrobials provide selective and mechanistically distinct alternatives to conventional antibiotics, but their performance in vivo is often limited by instability in physiological environments, immune neutralization, uneven tissue distribution, and insufficient access to bacteria protected by biofilms or surface-associated barriers. This narrative review examines how nanotechnology-based delivery systems can address these constraints. We first outline the delivery-relevant biological barrier characteristic of ESKAPE pathogens, then summarize the therapeutic potential and inherent limitations of whole phages, phage-derived enzymes, and CRISPR-based antimicrobials when used without formulation. Major nanotechnology platforms for antibacterial delivery are reviewed, followed by analysis of how nano-enabled systems can improve stability, localization, and persistence of these biological agents. A pathogen-aware integration framework is presented that links dominant barriers in each ESKAPE pathogen to the biological modality and nano-enabled delivery strategy most likely to enhance exposure at infection sites. Translational challenges, regulatory considerations, and emerging directions, including responsive delivery systems and personalized approaches, are also discussed. Overall, nano-enabled phage-based therapeutics represent a realistic and adaptable strategy for managing MDR ESKAPE infections. Therapeutic success depends on both continued discovery and engineering of antibacterial agents and effective delivery design.

Tuberculosis (TB) remains a leading infectious killer, increasingly complicated by multidrug-resistant (MDR) and extensively drug-resistant (XDR) disease; current regimens, although effective, are prolonged, toxic, and often fail to reach intracellular bacilli in heterogeneous lung lesions. This narrative review synthesizes how next-generation antimycobacterial strategies can be translated “from molecule to patient” by coupling potent therapeutics with delivery platforms tailored to the lesion microenvironment. We survey emerging small-molecule classes, including decaprenylphosphoryl-β-D-ribose 2′-epimerase (DprE1) inhibitors, mycobacterial membrane protein large 3 (MmpL3) inhibitors, and respiratory chain blockers, alongside optimized uses of established agents and host-directed therapies (HDTs). These are mapped to inhalable and nanocarrier systems that improve intralesional exposure, macrophage uptake, and targeted release while reducing systemic toxicity. Particular emphasis is placed on pulmonary dry powder inhalers (DPIs) and aerosols for direct lung targeting, stimuli-responsive carriers that trigger release through pH, redox, or enzymatic cues, and long-acting depots or implants that shift daily dosing to monthly or quarterly schedules to enhance adherence, safety, and access. We also outline translational enablers, including model-informed pharmacokinetic/pharmacodynamic (PK/PD) integration, device formulation co-design, manufacturability, regulatory quality frameworks, and patient-centered implementation. Overall, aligning stronger drugs with smart delivery platforms offers a practical pathway to shorter, safer, and more easily completed TB therapy, improving both individual outcomes and public health impact.

Food systems expose bacteria to repeated nonlethal stresses during primary production, processing, storage, and sanitation. Depending on the type, intensity, and sequence of exposure, these stresses may weaken cells, act synergistically to promote inactivation, or fail to eliminate contamination. Instead, they can alter bacterial physiology in ways that affect survival, recovery, detection, and responses to control measures. This review examines how stress history contributes to persistent food safety challenges. Listeria monocytogenes is used as a central biological model, with relevant comparisons to other foodborne pathogens. Evidence from food-processing and environmental studies shows that repeated sublethal stress can shift bacterial populations toward stress-hardened states. Here, “stress-hardened” refers to reversible physiological changes and the survival of more tolerant cells, not permanent genetic adaptation. These states include sublethal injury, delayed growth, viable but nonculturable cells, biofilm formation, and increased tolerance to later stresses. These outcomes contribute to, but do not fully explain, the persistence of L. monocytogenes in food environments; intrinsic traits such as psychrotrophic growth and interactions with endogenous microflora also play important roles. These factors help explain repeated recovery of L. monocytogenes after sanitation and the underestimation of viable cells by routine culture-based methods, which do not reliably indicate whether pre-stressed cells retain the potential to cause foodborne illness. Many monitoring and validation approaches rely on unstressed laboratory cultures and fixed enrichment protocols. These conditions do not reflect the physiological states encountered in real food systems. As a result, negative test results may reflect limited recovery rather than true absence, and control performance may be overestimated when stress-conditioned populations are not considered. Across the farm-to-fork continuum, stress responses, persistence mechanisms, and detection limitations are closely linked, indicating that stress history should be considered a core element of hazard characterization, monitoring, and control validation. Incorporating stress biology into food safety assessment can improve the realism of verification strategies when combined with risk characterization that considers infectious dose and host susceptibility, and support control strategies under real-world processing and environmental conditions.

Staphylococcus aureus (S. aureus) is one of the most prevalent bacterial pathogens recovered from diabetic foot infections (DFIs). Most S. aureus isolates exhibit methicillin resistance, so treatment is recommended with antimicrobials active against methicillin-resistant S. aureus (MRSA) in patients who have risk factors associated with MRSA infections. The main goal of this study was to see if proteomics and molecular methods could be effective in identifying and distinguishing MRSA recovered from DFIs. Since MRSA is highly resistant to β-lactam antibiotics and usually does not respond to other antimicrobial drugs, we evaluated the resistance of MRSA isolates against different antibiotics. The standard procedures were followed for a culture of 250 skin swabs collected from diabetic foot patients. The phenotypic characteristics of 48 suspected S. aureus cultures were determined via microscopic examination, Gram staining, a coagulase test, a BBL™ Staphyloslide™ Latex test, a Staph ID 32 API system, and a Vitek 2 Compact system. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to examine the protein profile of all isolates, and real-time PCR was then used to identify mecA and PVL virulence genes. S aureus isolates were tested using the Vitek 2 Compact for antimicrobial susceptibility using Gram-positive cards (GP71). Among the 48 bacterial isolates tested, 45 (93.75%), 42 (87.5%), and 46 (95.83%) were positive in tube coagulase, the Staph ID 32 API system, and the Vitek 2 Compact system, respectively. We correctly identified all suspected S. aureus isolates (100%) via MALDI-TOF MS with a score value ≥2.00 and differentiated them into 22/48 MRSA (45.83%) and 26/48 MSSA (54.17%) isolates. A higher peak intensity at masses of 5530 Da, 6580 Da, 6710 Da, and 6820 Da was detected in MRSA, but not in MSSA. All MRSA isolates tested positive for the mecA gene, while all isolates tested negative for the PVL gene. The antibiotic susceptibility results showed that 22 (100%), 20 (90.91%), 19 (86.36%), 18 (81.82%), 17 (77.27%), 15 (68.18%), 13 (59.1%), and 12 (54.55%) MRSA strains were resistant to cefoxitin, daptomycin, erythromycin, benzylpenicillin, ciprofloxacin, oxacillin, and clindamycin, respectively. In contrast, all MRSA strains were extremely susceptible (100%) to linezolid, nitrofurantoin, quinupristin–dalfopristin, tigecycline, and vancomycin. Moreover, 20 (90.91%), 18 (81.82%), and 17 (77.27%) of the MRSA strains exhibited high sensitivity against rifampin, trimethoprim–sulfamethoxazole, and gentamicin, respectively. In DFIs, MALDI-TOF MS is a powerful and accurate method of identifying and distinguishing both MRSA and MSSA isolates. A high level of antimicrobial resistance was found in MRSA isolates, and antibiotic therapy based on antibiotic susceptibility patterns is essential for a successful outcome.

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.

Healthcare-associated infections lead to considerable morbidity, a prolonged hospital stay, antibiotic resistance, long-term disability, mortality and increased healthcare costs. Based on the literature, some individual and socio-demographic factors including knowledge, age and length of service or work experience, gender and type of profession influence compliance with infection prevention and control procedures. In addition, organizational culture, which refers to the assumptions, values, and norms shared among colleagues, can influence an individual’s thinking and healthcare workers’ behavior, either positively or negatively. Infection control practices based on the perspective of patients, hospital management and healthcare workers may help develop a better understanding of the factors influencing compliance with infection prevention and control policies and guidelines.

Helicobacter pylori (H. pylori) is one of the most prevalent chronic bacterial infections globally, significantly contributing to gastritis, peptic ulcers, and gastric malignancies. Its pathogenesis involves a complex array of virulence factors—including cagA, vacA, and urease—which facilitate mucosal colonization, immune evasion, and persistent inflammation. A major challenge in vaccine development is the bacterium’s ability to manipulate both innate and adaptive immune responses, resulting in limited natural clearance and long-term persistence. This review synthesizes H. pylori pathogenesis and host immune dynamics, highlighting their implications for vaccine design. By elucidating the molecular and cellular mechanisms underlying host–pathogen interactions, we explore how these insights inform antigen selection, adjuvant optimization, and delivery strategies. By integrating basic science with translational objectives, this review aims to support the development of an effective H. pylori vaccine, addressing global health needs, particularly in regions with a high infection burden and limited access to treatment.