Acta Microbiologica Hellenica

The emergence and dissemination of antimicrobial resistance (AMR) are driven by complex, interconnected mechanisms involving microbial communities, environmental factors, and human activities, with climate change playing a pivotal and accelerating role. Rising temperatures, altered precipitation patterns, and other environmental disruptions caused by climate change create favorable conditions for bacterial growth and enhance the horizontal gene transfer (HGT) of antibiotic resistance genes (ARGs). Thermal stress and environmental pressures induce genetic mutations that promote resistance, while ecosystem disturbances facilitate the stabilization and spread of resistant pathogens. Moreover, climate change exacerbates public and animal health risks by expanding the range of infectious disease vectors and driving population displacement due to extreme weather events, further amplifying the transmission and evolution of resistant microbes. Livestock agriculture represents a critical nexus where excessive antibiotic use, environmental stressors, and climate-related challenges converge, fueling AMR escalation with profound public health and economic consequences. Environmental reservoirs, including soil and water sources, accumulate ARGs from agricultural runoff, wastewater, and pollution, enabling resistance spread. This review aims to demonstrate how the Mediterranean’s strategic position makes it an ideal living laboratory for the development of integrated “One Health” frameworks that address the mechanistic links between climate change and AMR. By highlighting these interconnections, the review underscores the need for a unified approach that incorporates sustainable agricultural practices, climate mitigation and adaptation within healthcare systems, and enhanced surveillance of zoonotic and resistant pathogens—ultimately offering a roadmap for tackling this multifaceted global health crisis. Full article

The European catfish (Silurus glanis, Linnaeus 1758), commonly known as the wels catfish, is one of the largest freshwater fish in Europe and an ecologically and economically important species in both natural ecosystems and aquaculture. Its broad native distribution, together with the rapid growth of farming practices, increases concerns about pathogen dissemination and their potential impact on biodiversity, animal health, and potential risks to human healthcare. This review is based on a structured literature search following PRISMA recommendations for narrative reviews and summarizes current knowledge on the main pathogen groups affecting S. glanis—viruses (ranaviruses, alloherpesviruses), bacteria (Aeromonas spp., Edwardsiella spp.), protozoan and metazoan parasites (Ichthyophthirius multifiliis, Thaparocleidus spp., Eustrongylides spp., Contracaecum larvae), and oomycetes (Saprolegnia spp., Branchiomyces spp.). Within the One Health approach, particular attention is given to zoonotic pathogens such as Aeromonas spp., Edwardsiella tarda, and helminths like Eustrongylides and Contracaecum, which may cause risks to human health through contaminated water or consumption of raw or undercooked fish. The review integrates findings from field surveys, regional case studies such as those from the Danube basin, and data from the authors’ doctoral research. Because the wels catfish is increasingly cultivated and serves as an apex predator in natural habitats, its effective disease management is critical for both aquaculture and wild populations, and also for the food chains at all. Strengthened surveillance, health monitoring, and biosecurity measures are essential preventing the introduction and spread of pathogens into new hosts and habitats. Through the underlining of major catfish pathogen groups, this review highlights key challenges within the One Health approach and underscores the need for integrated health monitoring, biosecurity, and environmental management strategies. Full article

Malassezia spp. has been recognized among neonatal intensive care unit (NICU) patients’ commensals and pathogens, accounting for a significant number of invasive fungal infections. The Random Amplified Polymorphic DNA (RAPD) may be used for Malassezia spp. strains typing from clinical isolates, demonstrating high resolution and specificity. Herein, we propose a retrospective analysis of Malassezia spp. isolates, aiming to investigate their identity and transmission pathways. Moreover, we documented Malassezia spp. prevalence within the University Hospital Policlinico of Catania, Italy. The analysis collected a total number of 16 M. pachydermatis and categorized them into four different clusters, hypothesizing a horizontal transmission. Although the essential role of microbiological sample cultures, our data suggested further environmental surveillance protocols to prevent NICU patients’ colonization due to the Malassezia spp. persistence and adhesion within healthcare surfaces. Full article

Linezolid (LZD) is an oxazolidinone antibiotic effective in the treatment of infection with Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). The decline in susceptibility to linezolid is a concern for antimicrobial chemotherapy. In this study, the prevalence of the LZD minimum inhibitory concentration (MIC) of 2 mg/L (LZD-MIC2), which represents a slightly high value within a range of susceptibility for S. aureus (≤4 mg/L), was investigated retrospectively for staphylococcal species from different sources. We collected the records of LZD MIC of Staphylococcus/Mammaliicoccus that had been obtained in our previous studies on isolates from patients, healthy individuals, and foodstuff. Prevalence of isolates showing LZD-MIC2 was analyzed depending on the type of staphylococcal species and S. aureus clones. In clinical isolates, methicillin-susceptible S. aureus (MSSA) and S. argenteus showed significantly higher LZD-MIC2 rates (20.0% and 21.5%, respectively) than MRSA (7.3%). Among clinical and colonizing isolates of MSSA, LZD-MIC2 was more commonly found in CC1 (ST188, ST2990, etc.), CC8, CC15, ST30, ST97, and ST121 than other lineages. In S. argenteus isolates, which were mostly methicillin-susceptible, there was no significant difference in the LZD-MIC2 prevalence among the three genotypes. The LZD-MIC2 was detected in 18.3% of coagulase-negative staphylococci (CoNS), with S. saprophyticus, S. pasteuri, and M. sciuri showing higher prevalence (30–57%) than other species. The present study revealed that the prevalence of the LZD-MIC2 is different depending on staphylococcal species/types, as they are more common in specific MSSA lineages and some CoNS species. Full article

West Nile Virus (WNV), a mosquito-borne flavivirus first identified in Uganda in 1937, has emerged over the past quarter century as a major global public health threat. Since its introduction into North America in 1999, WNV has become the leading cause of arboviral neuroinvasive disease, with recurrent outbreaks continuing across Europe, Africa, and the Americas. This review provides a comprehensive overview of the microbiology, epidemiology, and clinical impact of WNV. We discuss the molecular biology of the virus, highlighting its genomic organization, replication strategies, and the structural and non-structural proteins that underpin viral pathogenesis and immune evasion. The complex enzootic transmission cycle, involving Culex mosquitoes and diverse avian reservoir hosts, is examined alongside ecological and climatic determinants of viral amplification and spillover into humans and equines. The clinical spectrum of WNV infection is outlined, ranging from asymptomatic seroconversion to West Nile fever and life-threatening neuroinvasive disease, with particular emphasis on risk factors for severe outcomes in older and immunocompromised individuals. Current approaches to diagnosis, supportive management, and vector control are critically reviewed, while challenges in vaccine development and the absence of effective antiviral therapy are underscored. Finally, we address future research priorities, including therapeutic innovation, predictive outbreak modeling, and genomic surveillance of viral evolution. WNV exemplifies the dynamics of emerging zoonotic diseases, and its persistence underscores the necessity of a coordinated One Health approach integrating human, animal, and environmental health. Continued scientific advances and public health commitment remain essential to mitigate its enduring global impact. Full article

Severely burned patients are at high risk of local and systemic infections due to skin barrier loss. Their clinical management is complex and requires coordinated intensive care and infection prevention strategies. Diagnosing infective endocarditis (IE) in this population is particularly difficult due to overlapping symptoms and limited diagnostic specificity. Common pathogens include Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii. We conducted a retrospective cohort study on 543 patients with burns affecting >18% of total body surface area (TBSA), admitted to our Burn Intensive Care Unit (BICU) from 2019 to 2024. The incidence of infective endocarditis was 1.47%, involving aortic (75%), mitral (12.5%), and tricuspid (12.5%) valves. Pathogens identified included S. aureus, Klebsiella pneumoniae, A. baumannii, and P. aeruginosa. This incidence is significantly higher than that in the general population. Mortality reached 50%, with an overall 3-month mortality of 75%. The literature on IE in burn patients is scarce, and the role of antibiotic prophylaxis remains controversial. Infective endocarditis in burn patients, although rare, represents a severe complication with high mortality. Early diagnosis and coordinated multidisciplinary care are essential to improve patient outcomes. Full article

The COVID-19 pandemic resulted in high morbidity and mortality, as well as severe social and economic disruption globally. Since the pandemic began in 2019, the severe acute respiratory syndrome, coronavirus 2, has undergone numerous changes, resulting in the emergence of new variants and subvariants. The emergence of new variants of the virus poses a challenge to scientists. There is currently no SARS-CoV-2 variant meeting the criteria of variants of concern, whereas the only variant of interest is JN.1, and there are six variants under monitoring: LP8.1, NP1.8.1, XEC, KP.3, KP.3.1.1 and the latest, XFG (Stratus). Although the latter appears to be more transmissible than the others, genomic evidence indicates that it is less aggressive than some recent variants. Nevertheless, continuous genomic surveillance of COVID-19 is still important to detect any new variants that could threaten public health. Numerous therapeutic strategies, such as drugs, vaccines, and nutritional supplements, are being used to treat COVID-19. This narrative review is an overview of COVID-19 and its various facets, from the number of cases to the therapies used, the current variants, and the ongoing clinical trials, specifically focusing on the most recent studies. Full article

Genome editing technologies, including CRISPR/Cas9, TALENs, and ZFNs, offer promising approaches to disrupt HPV oncogenes E6 and E7, thereby restoring tumor-suppressor pathways. In this review, we summarize recent preclinical findings demonstrating selective apoptosis and tumor regression in HPV-positive cell and animal models, as well as early-phase clinical studies exploring local CRISPR-based therapies. We also compare the relative strengths and limitations of major editing platforms, discuss delivery strategies, and highlight their potential integration with immunotherapy and conventional treatments. While preclinical studies show encouraging efficacy (e.g., up to 60% tumor regression in xenograft models and marked reactivation of p53/pRb pathways), translation into routine practice remains limited by challenges such as efficient delivery, minimizing off-target effects, long-term safety, cost, and ethical considerations. Continued optimization of high-fidelity nucleases, tissue-specific delivery systems, and genotype-tailored guide RNAs will be essential. Genome editing therefore represents a potential future addition to the therapeutic landscape of HPV-related diseases, but substantial barriers must be addressed before clinical implementation. Full article

Dermatophilosis, caused by the Gram-positive, filamentous bacterium Dermatophilus congolensis, is an important skin disease that adversely affects cattle health and productivity. It also affects other domestic and wild animals and occasionally humans. This review provides a detailed overview of the molecular characteristics and resistome profile of D. congolensis, highlighting recent advances in genomic research. We examine the bacterium’s genome architecture, including its genome size, GC content, gene composition, and phylogenetic placement within the Actinomycetales. Key virulence factors are discussed, including proteolytic enzymes, hyphal invasion, zoospore motility, and the gene products of nasp and agac, emphasizing their roles in tissue invasion, pathogenesis, and diagnostic detection. Furthermore, we analyze resistome, focusing on identified antibiotic resistance genes, diverse resistance mechanisms such as efflux pumps and beta-lactamases, and the contribution of mobile genetic elements to horizontal gene transfer. The implications of these molecular insights for accurate diagnosis, effective treatment, and antibiotic stewardship in cattle production are critically evaluated. Finally, we highlight future research priorities aimed at deepening our understanding of D. congolensis biology and improving strategies for disease control. This review underscores the importance of integrating molecular surveillance with antimicrobial monitoring to safeguard cattle health and promote sustainable livestock management. Full article

Worldwide, polymicrobial infections (PMIs) account for an estimated 20–50% of severe clinical infection cases, with biofilm-associated and device-related infections reaching 60–80% in hospitalized patients. This review discusses the clinical burden of major infections in which PMIs are almost inevitable, such as diabetic foot infections, intra-abdominal infections, pneumonia, and biofilm-associated device infections. Globally, the PMI landscape is diverse; however, the Indian subcontinent is a PMI hotspot where high comorbidities, endemic antimicrobial resistance, and underdeveloped diagnostic capacity elevate the risks of poor outcomes. Existing diagnostic like culture-based methods, PCR panels, sequencing, and biomarker-based assays are constrained by sensitivity, turnaround times (TATs), and high costs. Vulnerable populations, particularly neonates, the elderly, immunocompromised patients, and socioeconomically marginalized groups, show case-fatality rates 2-fold higher than monomicrobial infections in similar settings. Emerging diagnostic solutions include CRISPR-based multiplex assays, artificial intelligence-based metagenomic platforms, and sensitive biosensors with point-of-care applicability. These technologies show potential in reducing the TAT (<2 h) with high accuracy (>95%). However, their translation to real-world settings depends critically on affordability, integration into healthcare pathways, and supportive policy. This will provide equitable diagnostic access, particularly in low- and middle-income countries (LMICs). Full article

Age-related impaired wounds often become infected with bacteria, leading to substantial mortality and morbidity in the elderly. The decline in androgen levels with increasing age is believed to exacerbate inflammation during wound infections. Despite its well-documented anti-inflammatory activities in wound repair, little is known about the effect of age-related androgen deprivation on bacterial phagocytosis in impaired chronic wounds. The aim of this study was to investigate the effect of age-related testosterone deprivation on the phagocytic functions of THP-1 monocyte-derived macrophages to eliminate Gram-positive and Gram-negative bacteria in vitro. Host–pathogen interaction experiments were conducted to quantify the macrophage-mediated clearance of two common wound-associated bacteria, methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa, under in vitro environments that model testosterone levels representative of those found in elderly males, healthy young adults and testosterone replacement therapy (TRT). Testosterone and its metabolite 5α-dihydrotestosterone (DHT) significantly dampened the macrophage-mediated phagocytosis of both MRSA and P. aeruginosa in a dose-dependent manner (p < 0.05). Blockade of the androgen receptor (AR) using enzalutamide confirmed that testosterone mediates bacterial clearance through binding to the AR. Blocking the conversion of testosterone to DHT through stimulation of macrophages with the 5-α-reductase inhibitor finasteride reversed the testosterone-mediated effects on bacterial clearance, which confirmed that testosterone could potentially dampen the innate phagocytic responses in macrophages through conversion to DHT. Novel findings in this study suggest that the selective manipulation of the AR and/or blockade of testosterone–DHT conversion may provide effective therapeutic treatments to combat wound infections in the elderly. Full article

Stenotrophomonas maltophilia is a nosocomial pathogen that is resistant to many broad-spectrum antibiotics. This study aimed to evaluate the in vitro synergy of trimethoprim/sulfamethoxazole (SXT) combined with levofloxacin (LEV) or ceftazidime (CAZ) using checkerboard and gradient diffusion methods. Between 2016 and 2021, 20 S. maltophilia strains (five SXT-resistant and 15 SXT-susceptible strains) were collected from various clinical settings. Their susceptibility to SXT, LEV, and CAZ was assessed using both checkerboard and gradient diffusion synergy tests. The gradient diffusion method was performed using commercial strip-based tests (Liofilchem^®). It should be noted that the gradient diffusion method has not been standardized by the Clinical and Laboratory Standards Institute (CLSI) for synergy testing and is considered for research purposes only. In the checkerboard method, the SXT + LEV combination showed synergy in one strain and an additive effect in 19 strains; the SXT + CAZ combination exhibited synergy in eight strains and an additive effect in 12 strains. In the gradient diffusion method, the SXT + LEV combination showed synergy in one strain and an additive effect in 19 strains; the SXT + CAZ combination exhibited synergy in five strains, an additive effect in 14 strains, and antagonism in one strain. A correlation between the two methods was observed in 90% of SXT + LEV combinations and 65% of SXT + CAZ combinations. Both checkerboard and gradient diffusion methods yielded similar results, indicating their reliability in determining antibiotic combinations. Given the observed synergy, CAZ combinations may be effective for treating SXT-resistant strains. Full article

Microbiome-targeted therapies are redefining gastroenterology by delivering precision interventions that align with the body’s natural microbial ecosystem. This narrative review evaluates evidence for established approaches, probiotics, prebiotics, fecal microbiota transplantation (FMT), and postbiotics, and examines emerging innovations such as engineered probiotics, bacteriophage therapy, and metabolite-based interventions. Cure rates for recurrent Clostridium difficile infection in randomized trials range from 67% to 94%, depending on route and donor protocol, while multi-strain probiotics provide moderate benefits in inflammatory bowel disease. New modalities, including engineered bacteria and defined bacterial consortia, have progressed to Phase 3 trials, with several granted FDA breakthrough therapy designation. Approvals of Rebyota and Vowst mark a pivotal milestone, creating validated regulatory pathways for microbiome therapeutics. Despite progress, challenges remain in protocol standardisation, patient selection, cost-effectiveness, and clinical integration. Over 200 active trials and growing pharmaceutical investment signal a robust pipeline, with applications expanding to oncology, metabolic disorders, and immune modulation. Continued progress depends on validated biomarkers and personalized strategies guided by microbiome profiling. International regulatory harmonization will also be required to ensure safe and equitable adoption. The field is shifting toward working with, rather than against, the body’s microbial ecosystem, offering substantial potential for personalized gastrointestinal disease management. Full article

Background: Pancreatic stone protein (PSP) has emerged as a promising biomarker for the early diagnosis of sepsis, infectious diseases, and chronic inflammatory conditions, including cancer, diabetes, and inflammatory bowel disease. This review evaluates the role of PSP as a biomarker and functional protein, emphasizing its diagnostic and prognostic value. Methods: This review was conducted through a comprehensive literature search using the PubMed database, covering publications from the discovery of PSP in the 1980s up to 2025. The information was gathered into thematic sections to provide a comprehensive and structured review of PSP as a biomarker and functional protein in health and disease. Results: A total of 256 articles were reviewed and critically assessed, with 80 meeting the inclusion criteria and forming the basis of this review. The reviewed literature underscores PSP as a multifaceted protein involved in inflammation, immune modulation, tissue regeneration, and systemic infection. PSP shows promise as an early biomarker in critically ill patients. Conclusions: PSP is a clinically promising but not yet fully validated biomarker. To enable its routine clinical application, standardized diagnostic cutoffs, validation through multicenter trials, and integration with existing biomarker panels are required. Further research is warranted to fully establish its diagnostic and prognostic potential across diverse clinical settings. Full article

Aeromonas is a genus of Gram-negative, facultatively anaerobic bacteria commonly found in aquatic environments and increasingly recognized as opportunistic pathogens. Among them, Aeromonas dhakensis stands out for its high virulence and antimicrobial resistance, but it is often misidentified due to its phenotypic similarity with A. hydrophila and A. caviae. In this study, a microorganism was isolated from the peritoneal fluid of a patient with signs of intra-abdominal infection. MALDI-TOF MS initially suggested A. hydrophila or A. caviae, but the identification was confirmed with high confidence only after further molecular analyses and the use of genome-based tools, which identified the organism as A. dhakensis. This was further supported by phylogenomic and ANI analysis. Resistome analysis revealed intrinsic resistance genes, including a chromosomal class C β-lactamase and an imiH-type carbapenemase, consistent with the observed carbapenem resistance. No plasmid-mediated carbapenemases were found. These findings underscore the limitations of MALDI-TOF in identifying Aeromonas species in certain cases and highlight the value of genomic approaches for accurate species determination and resistance profiling, especially for isolates from sterile body fluids. Full article

Sexually transmitted infections (STIs) are a significant global health concern, affecting millions of people worldwide, particularly sexually active adolescents and young adults. These infections, caused by various pathogens, including bacteria, viruses, parasites, and fungi, can have profound implications for women’s reproductive health and fertility. This review explores the role of vaginal and uterine infections in women’s infertility, focusing on the most common pathogens and their impact on reproductive outcomes. Bacterial infections, such as those caused by intracellular bacteria (Mycoplasma, Ureaplasma, and Chlamydia), Neisseria gonorrhoeae, and bacterial vaginosis, are among the most prevalent causes of infertility in women. Studies have shown that these infections can lead to pelvic inflammatory disease, tubal occlusion, and endometrial damage, all of which can impair fertility. Mycobacterium tuberculosis, in particular, is a significant cause of genital tuberculosis and infertility in high-incidence countries. Viral infections, such as Human papillomavirus (HPV) and Herpes simplex virus (HSV), can also affect women’s fertility. While the exact role of HPV in female infertility remains unclear, studies suggest that it may increase the risk of endometrial implantation issues and miscarriage. HSV may be associated with unexplained infertility. Parasitic infections, such as trichomoniasis and schistosomiasis, can directly impact the female reproductive system, leading to infertility, ectopic pregnancy, and other complications. Fungal infections, such as candidiasis, are common but rarely have serious outcomes related to fertility. The vaginal microbiome plays a crucial role in maintaining reproductive health, and alterations in the microbial balance can increase susceptibility to STIs and infertility. Probiotics have been proposed as a potential therapeutic strategy to restore the vaginal ecosystem and improve fertility outcomes, although further research is needed to establish their efficacy. In conclusion, vaginal and uterine infections contribute significantly to women’s infertility, with various pathogens affecting the reproductive system through different mechanisms. Early diagnosis, appropriate treatment, and preventive measures are essential to mitigate the impact of these infections on women’s reproductive health and fertility. Full article

This study assessed microbiological contamination in street-sold meat products, focusing on Enterobacterales and coagulase-negative staphylococci (CoNS) species and their antibiotic resistance. Chicken and mutton street foods like shawarma and brochettes were tested for bacterial load, species distribution. and resistance profiles. The results showed significant contamination, with Enterobacter cloacae (5.38 Log 10 CFU/g). Staphylococcus lentus and Staphylococcus xylosus were also common, reaching 6.23 Log 10 CFU/g in some samples. Contamination levels varied significantly by food type, with chicken shawarma showing the highest risk. Antimicrobial susceptibility testing revealed high multidrug resistance, particularly among E. cloacae and Staphylococcus species. Biofilm formation an indicator of resistance was observed mainly in staphylococci and enhanced under fed-batch culture. These findings highlight public health concerns tied to poor hygiene and undercooking in street food environments. The study emphasizes the need for improved hygiene practices, standardized cooking methods, and systematic food safety monitoring to reduce contamination and antibiotic resistance risks. Full article

Syphilis, which is caused by Treponema pallidum, remains one of the most common congenital infection worldwide and has tremendous consequences for the mother and her developing foetus if left untreated. The complexity of the exposure to this pathogen extends beyond the well-established clinical manifestations, as it can profoundly affect placental histomorphology. This study aimed to compare T. pallidum-exposed placental villi structures with healthy placentae at term to evaluate the histomorphological differences using stereology. In this case-control study conducted at term (38 weeks ± 2 weeks), 78 placentae were collected from the hospital delivery suites, comprising 39 cases (T. pallidum-exposed) and 39 controls (non-exposed), who were gestational age-matched with other potential confounders excluded. Blood samples from the umbilical vein and placental basal plate were tested for syphilis, using rapid diagnostic test (RDT) kits for T. pallidum (TP) antibodies (IgG and IgM) to classify placentae as exposed to T. pallidum (cases) and non-exposed (controls). Tissue sections were prepared and stained with haematoxylin and eosin, and the mean volume densities of syncytial knots, foetal capillaries, syncytial denuded areas, and intervillous spaces were estimated using stereological methods. Statistical analysis was performed to compare the mean values between the case and control groups. Stereological assessment revealed significant differences between the T. pallidum-exposed and non-exposed groups with regard to syncytial knots (p < 0.0001), syncytial denudation (p < 0.0001), and foetal capillaries (p < 0.0001), but no significant difference in the intervillous space was found (p = 0.1592). Therefore, our study shows, for the first time, that the histomorphology of human placental villi appears to be altered by exposure to T. pallidum. It will, therefore, be interesting to determine whether these changes in the placental villi translate into long-term effects on the baby. Full article

The excessive use of antimicrobials drives the emergence of multidrug resistance (MDR) in bacterial strains, which harbor resistance genes to survive under diverse drug pressures. Such resistance can result in life-threatening infections. The predominance of MDR Pseudomonas spp. poses significant challenges to public health and environmental sustainability, particularly in ecosystems affected by human activities. Characterizing MDR Pseudomonas spp. is crucial for developing effective diagnostic tools and biosecurity protocols, with broader implications for managing other pathogenic bacteria. Strains were diagnosed through 16S rRNA PCR and sequencing, complemented by phylogenetic analysis to evaluate local and global evolutionary connections. Antibiotic susceptibility tests revealed extensive resistance across multiple classes, with MIC values surpassing clinical breakpoints. This study examined the genetic diversity, resistance potential, and phylogenetic relationships among Pseudomonas aeruginosa strain DG2 and Pseudomonas fluorescens strain FM3, which were isolated from solid waste dump sites (n = 30) and dairy farms (n = 22) in West Bengal, India. Phylogenetic analysis reveals distinct clusters that highlight significant geographic linkages and genetic variability among the strains. Significant biofilm production under antibiotic exposure markedly increased resistance levels. RAPD-PCR profiling revealed substantial genetic diversity among the isolates, indicating variations in their genetic makeup. In contrast, SDS-PAGE analysis provided insights into the protein expression patterns that are activated by stress, which are closely linked to MDR. This dual approach offers a clearer perspective on their adaptive responses to environmental stressors. This study underscores the need for vigilant monitoring of MDR Pseudomonas spp. in anthropogenically impacted environments to mitigate risks to human and animal health. Surveillance strategies combining phenotypic and molecular approaches are essential to assess the risks posed by resilient pathogens. Solid waste and dairy farm ecosystems emerge as critical reservoirs for the evolution and dissemination of MDR Pseudomonas spp. Full article