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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.

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.

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.

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.

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) infection, which affects approximately half of the world’s population, remains a serious public health problem. As H. pylori infection leads to a number of gastric pathologies, including inflammation, gastroduodenal ulcers, and malignancies, early detection and treatment are crucial to preventing the spread of the infection. Multiple extragastric complications, such as iron deficiency anaemia, immune thrombocytopenic purpura, vitamin B12 deficiency, diabetes mellitus, cardiovascular diseases, and certain neurological disorders, have also been linked to H. pylori infection. An awareness of H. pylori and associated health hazards is necessary to minimize or even eradicate the infection. Therefore, there is an urgent need to raise the standards for the currently employed diagnostic, eradication, alternative treatment strategies. In addition, a brief overview of traditional and cutting-edge approaches that have proven effective in identifying and managing H. pylori is needed. Based on the test and laboratory equipment available and patient clinical characteristics, the optimal diagnostic approach requires weighing several factors. The pathophysiology and pathogenic mechanisms of H. pylori should also be studied, focusing more on the infection-causing virulence factors of this bacterium. Accordingly, this review aims to demonstrate the various diagnostic, pathophysiological, therapeutic, and eradication tactics available for H. pylori, emphasizing both their advantages and disadvantages. Invasive methods (such as quick urease testing, biopsy, or culture) or noninvasive methods (such as breath tests, stool investigations, or serological tests) can be used. We also present the most recent worldwide recommendations along with scientific evidence for treating H. pylori. In addition to the current antibiotic regimens, alternative therapies may also be considered. It is imperative to eradicate the infections caused by H. pylori as soon as possible to prevent problems and the development of stomach cancer. In conclusion, significant advances have been made in identifying and treating H. pylori. To improve eradication rates, peptide mass fingerprinting can be used as a diagnostic tool, and vaccines can also eliminate the infection.

Pathogens found in food are believed to be the leading cause of foodborne illnesses; and they are considered a serious problem with global ramifications. During the last few decades, a lot of attention has been paid to determining the microorganisms that cause foodborne illnesses and developing new methods to identify them. Foodborne pathogen identification technologies have evolved rapidly over the last few decades, with the newer technologies focusing on immunoassays, genome-wide approaches, biosensors, and mass spectrometry as the primary methods of identification. Bacteriophages (phages), probiotics and prebiotics were known to have the ability to combat bacterial diseases since the turn of the 20th century. A primary focus of phage use was the development of medical therapies; however, its use quickly expanded to other applications in biotechnology and industry. A similar argument can be made with regards to the food safety industry, as diseases directly endanger the health of customers. Recently, a lot of attention has been paid to bacteriophages, probiotics and prebiotics most likely due to the exhaustion of traditional antibiotics. Reviewing a variety of current quick identification techniques is the purpose of this study. Using these techniques, we are able to quickly identify foodborne pathogenic bacteria, which forms the basis for future research advances. A review of recent studies on the use of phages, probiotics and prebiotics as a means of combating significant foodborne diseases is also presented. Furthermore, we discussed the advantages of using phages as well as the challenges they face, especially given their prevalent application in food safety.

Healthcare-associated infections (HAIs) lead to considerable morbidity. Prolonged hospital HAIs, also known as hospital infections or nosocomial infections, refer to infectious diseases that occur within 48 h of hospital admission, within three days of discharge or 30 days after having received healthcare. A total of 39 government-owned hospitals with a total of 10,822 beds providing the majority (59.9%) of Saudi Arabian healthcare were analyzed. Medicare and Medicaid use hospital data to track hospital performance on matters pertaining to IPC. In addition, many hospitals have limited capacity with which to develop the core components required to build an effective IPC programme. Hajj has been associated with the increased prevalence and spread of infectious diseases. These factors include changes in human demographics and behaviour, the effect of new technologies and industries, an increase in international travel and commerce, and breakdown resulting from public health measures. Overall public health service expenditure originates from the administration and services that are provided free of charge to all Saudi citizens.

Brucellae are intracellular sneaky bacteria and they can elude the host’s defensive mechanisms, resulting in therapeutic failure. Therefore, the goal of this investigation was to rapid identification of Brucella species collected from animals and humans in Saudi Arabia, as well as to evaluate their resistance to antibiotics. On selective media, 364 animal samples as well as 70 human blood samples were cultured. Serological and biochemical approaches were initially used to identify a total of 25 probable cultured isolates. The proteomics of Brucella species were identified using the MALDI Biotyper (MBT) system, which was subsequently verified using real-time polymerase chain reaction (real-time PCR) and microfluidic electrophoresis assays. Both Brucella melitensis ( B . melitensis ) and Brucella abortus ( B . abortus ) were tested for antimicrobial susceptibility using Kirby Bauer method and the E-test. In total, 25 samples were positive for Brucella and included 11 B . melitensis and 14 B . abortus isolates. Twenty-two out of 25 (88%) and 24/25 (96%) of Brucella strains were recognized through the Vitek 2 Compact system. While MBT was magnificently identified 100% of the strains at the species level with a score value more than or equal to 2.00. Trimethoprim-sulfamethoxazole, rifampin, ampicillin-sulbactam, and ampicillin resistance in B . melitensis was 36.36%, 31.82%, 27.27%, and 22.70%, respectively. Rifampin, trimethoprim-sulfamethoxazole, ampicillin, and ampicillin-sulbactam resistance was found in 35.71%, 32.14%, 32.14%, and 28.57% of B . abortus isolates, correspondingly. MBT confirmed by microfluidic electrophoresis is a successful approach for identifying Brucella species at the species level. The resistance of B . melitensis and B . abortus to various antibiotics should be investigated in future studies.